Production of recombinant lytic polysaccharide monooxygenases and evaluation effect of its addition into Aspergillus fumigatus var. niveus cocktail for sugarcane bagasse saccharification.

Lignocellulosic biomass is a promising alternative for producing biofuels, despite its recalcitrant nature. There are microorganisms in nature capable of efficiently degrade biomass, such as the filamentous fungi. Among them, Aspergillus fumigatus var. niveus (AFUMN) has a wide variety of carbohydrate-active enzymes (CAZymes), especially hydrolases, but a low number of oxidative enzymes in its genome. To confirm the enzymatic profile of this fungus, this study analyzed the secretome of AFUMN cultured in sugarcane bagasse as the sole carbon source. As expected, the secretome showed a predominance of hydrolytic enzymes compared to oxidative activity. However, it is known that hydrolytic enzymes act in synergy with oxidative proteins to efficiently degrade cellulose polymer, such as the Lytic Polysaccharide Monooxygenases (LPMOs). Thus, three LPMOs from the fungus Thermothelomyces thermophilus (TtLPMO9D, TtLPMO9H, and TtLPMO9O) were selected, heterologous expressed in Aspergillus nidulans, purified, and used to supplement the AFUMN secretome to evaluate their effect on the saccharification of sugarcane bagasse. The saccharification assay was carried out using different concentrations of AFUMN secretome supplemented with recombinant T. thermophilus LPMOs, as well as ascorbic acid as reducing agent for oxidative enzymes. Through a statistic design created by Design-Expert software, we were able to analyze a possible cooperative effect between these components. The results indicated that, in general, the addition of TtLPMO9D and ascorbic acid did not favor the conversion process in this study, while TtLPMO9O had a highly significant cooperative effect in bagasse saccharification compared to the control using only AFUMN secretome.

Consumption of free sugars and dietary sources in the diet of university students according to socioeconomic and lifestyle factors / Consumo de açúcar livre e fontes dietéticas na dieta de estudantes universitários segundo fatores socioeconômicos e de estilo de vida

ABSTRACT Objective To evaluate the consumption of free sugars and associated factors in the diet of university students entering full-time courses at a public university in Mato Grosso. Methods Cross-sectional study with students aged 16 to 26 years in the first semester of 2016, 2017, and 2018. Average sugar consumption was estimated using a 24-hour diet recall, evaluating the percentage of free sugars in total energy intake and the prevalence of consumption greater than 10.0% of total energy intake, according to socioeconomic and lifestyle variables, stratified by sex, and the foods that most contributed to its consumption. Results A total of 1,063 students were evaluated, the percentage of free sugars in the students' diet was on average 14.3% among men and 15.4% among women, with the prevalence of consumption above 10.0% also being higher among women (72.6 vs. 66.6%). There was a direct relationship between the participation of free sugars in the highest tertiles of energy intake, and the main food groups sources of sugar were desserts, juices, soft drinks, and coffee. There was a significant association between courses outside the health sciences, tobacco use, and unsatisfactory meal consumption profile with higher consumption of free sugars among women. Among men, sugar consumption was higher among those who were not overweight compared to those who were overweight. Conclusion The consumption of free sugars is high among university students, being associated with higher energy consumption, especially for sweetened beverages.

RESUMO Objetivo Avaliar o consumo de açúcar livre e os fatores associados na alimentação de universitários ingressantes em cursos de período integral em uma universidade pública de Mato Grosso. Métodos Estudo transversal com estudantes de 16 a 26 anos de idade no primeiro semestre de 2016, 2017 e 2018 (n=1.063). O consumo médio de açúcar foi estimado por meio de recordatório de 24 horas, sendo avaliada a participação do açúcar livre na ingestão energética total e a prevalência de participação do consumo superior a 10,0% da ingestão energética, segundo variáveis socioeconômicas, de estilo de vida e fatores dietéticos, , estratificadas por sexo. Resultados A participação do açúcar livre na dieta foi, em média, de 14,3% entre os homens e 15,4% entre as mulheres, sendo a prevalência de consumo acima de 10,0% também maior entre as mulheres (72,6 vs. 66,6%). Foi maior a participação do açúcar livre nos tercis mais elevados de ingestão energética, sendo verificado que os principais grupos de alimentos fontes foram sobremesas, sucos, refrigerantes e café. Entre as mulheres, verificou-se maior consumo de açúcar livre entre aquelas que faziam uso de tabaco e que apresentavam perfil de consumo de refeições insatisfatório. Por outro lado, menor consumo de açúcar livre foi observado entre as estudantes da área da saúde. Entre os homens, o consumo de açúcar livre foi maior entre aqueles sem excesso de peso, comparados aos com excesso. Conclusão O consumo de açúcar livre foi elevado entre estudantes universitários, sendo maior entre os estudantes com maior consumo energético. Verificou-se que as bebidas adoçadas estiveram entre as principais fontes.

Heme-deficient metabolism and impaired cellular differentiation as an evolutionary trade-off for human infectivity in Trypanosoma brucei gambiense.

Resistance to African trypanosomes in humans relies in part on the high affinity targeting of a trypanosome lytic factor 1 (TLF1) to a trypanosome haptoglobin-hemoglobin receptor (HpHbR). While TLF1 avoidance by the inactivation of HpHbR contributes to Trypanosoma brucei gambiense human infectivity, the evolutionary trade-off of this adaptation is unknown, as the physiological function of the receptor remains to be elucidated. Here we show that uptake of hemoglobin via HpHbR constitutes the sole heme import pathway in the trypanosome bloodstream stage. T. b. gambiense strains carrying the inactivating mutation in HpHbR, as well as genetically engineered T. b. brucei HpHbR knock-out lines show only trace levels of intracellular heme and lack hemoprotein-based enzymatic activities, thereby providing an uncommon example of aerobic parasitic proliferation in the absence of heme. We further show that HpHbR facilitates the developmental progression from proliferating long slender forms to cell cycle-arrested stumpy forms in T. b. brucei. Accordingly, T. b. gambiense was found to be poorly competent for slender-to-stumpy differentiation unless a functional HpHbR receptor derived from T. b. brucei was genetically restored. Altogether, we identify heme-deficient metabolism and disrupted cellular differentiation as two distinct HpHbR-dependent evolutionary trade-offs for T. b. gambiense human infectivity.

The translational challenge in Chagas disease drug development.

Chagas disease is a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi. There is an urgent need for safe, effective, and accessible new treatments since the currently approved drugs have serious limitations. Drug development for Chagas disease has historically been hampered by the complexity of the disease, critical knowledge gaps, and lack of coordinated R&D efforts. This review covers some of the translational challenges associated with the progression of new chemical entities from preclinical to clinical phases of development, and discusses how recent technological advances might allow the research community to answer key questions relevant to the disease and to overcome hurdles in R&D for Chagas disease.

Association among gestational diabetes mellitus, periodontitis and prematurity: a cross-sectional study.

Objective: Gestational diabetes mellitus (GDM) causes maternal and infant morbidity. Periodontitis is associated with adverse pregnancy outcomes. The aim of this study was to evaluate periodontal status, prematurity and associated factors in pregnant women with and without GDM. Methods: This observational cross-sectional study included 80 pregnant women with GDM (G1 = 40) and without GDM (G2 = 40). Demographic and socioeconomic status, systemic and periodontal health condition, prematurity and newborns' birth weight were analyzed. For bivariate analysis, Mann-Whitney U-test, t test and Chi-squared test were used. Binary logistic regression analyzed independent variables for periodontitis and prematurity (p < 0.05). Results: Patients from G1 presented lower socioeconomic status, higher weight and body mass index (BMI). Prematurity (G1 = 27.5%; G2 = 2.5%; p < 0.05) and severe periodontitis percentages (G1 = 22.5%; G2 = 0; p = 0.001) were higher in G1 than in G2. Logistic regression analysis showed that household monthly income (OR = 0.65; 95% CI 0.48-0.86; p = 0.003) and maternal BMI (adjusted OR = 1.12; 95% CI 1.01-1.25; p = 0.028) were significant predictors of periodontitis during the third trimester of pregnancy. Presence of GDM remained in the final logistic model related to prematurity (adjusted OR = 14.79; 95% CI 1.80-121.13; p = 0.012). Conclusion: Pregnant women with GDM presented higher severity of periodontitis, lower socioeconomic status, higher overweight/obesity and a 10-fold higher risk of prematurity. Socioeconomic-cultural status and BMI were significant predictors for periodontitis, and GDM was a predictor to prematurity.

Association among gestational diabetes mellitus, periodontitis and prematurity: a cross-sectional study

ABSTRACT Objective: Gestational diabetes mellitus (GDM) causes maternal and infant morbidity. Periodontitis is associated with adverse pregnancy outcomes. The aim of this study was to evaluate periodontal status, prematurity and associated factors in pregnant women with and without GDM. Subjects and methods: This observational cross-sectional study included 80 pregnant women with GDM (G1 = 40) and without GDM (G2 = 40). Demographic and socioeconomic status, systemic and periodontal health condition, prematurity and newborns' birth weight were analyzed. For bivariate analysis, Mann-Whitney U-test, t test and Chi-squared test were used. Binary logistic regression analyzed independent variables for periodontitis and prematurity (p < 0.05). Results: Patients from G1 presented lower socioeconomic status, higher weight and body mass index (BMI). Prematurity (G1 = 27.5%; G2 = 2.5%; p < 0.05) and severe periodontitis percentages (G1 = 22.5%; G2 = 0; p = 0.001) were higher in G1 than in G2. Logistic regression analysis showed that household monthly income (OR = 0.65; 95% CI 0.48-0.86; p = 0.003) and maternal BMI (adjusted OR = 1.12; 95% CI 1.01-1.25; p = 0.028) were significant predictors of periodontitis during the third trimester of pregnancy. Presence of GDM remained in the final logistic model related to prematurity (adjusted OR = 14.79; 95% CI 1.80-121.13; p = 0.012). Conclusions: Pregnant women with GDM presented higher severity of periodontitis, lower socioeconomic status, higher overweight/obesity and a 10-fold higher risk of prematurity. Socioeconomic-cultural status and BMI were significant predictors for periodontitis, and GDM was a predictor to prematurity.

The translational challenge in Chagas disease drug development

Chagas disease is a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi. There is an urgent need for safe, effective, and accessible new treatments since the currently approved drugs have serious limitations. Drug development for Chagas disease has historically been hampered by the complexity of the disease, critical knowledge gaps, and lack of coordinated R&D efforts. This review covers some of the translational challenges associated with the progression of new chemical entities from preclinical to clinical phases of development, and discusses how recent technological advances might allow the research community to answer key questions relevant to the disease and to overcome hurdles in R&D for Chagas disease.

Cyclophilin 19 secreted in the host cell cytosol by Trypanosoma cruzi promotes ROS production required for parasite growth.

Infection by Trypanosoma cruzi, the protozoan parasite that causes Chagas disease, depends on reactive oxygen species (ROS), which has been described to induce parasite proliferation in mammalian host cells. It is unknown how the parasite manages to increase host ROS levels. Here, we found that intracellular T. cruzi forms release in the host cytosol its major cyclophilin of 19 kDa (TcCyp19). Parasites depleted of TcCyp19 by using CRISPR/Cas9 gene replacement proliferate inefficiently and fail to increase ROS, compared to wild type parasites or parasites with restored TcCyp19 gene expression. Expression of TcCyp19 in L6 rat myoblast increased ROS levels and restored the proliferation of TcCyp19 depleted parasites. These events could also be inhibited by cyclosporin A, (a cyclophilin inhibitor), and by polyethylene glycol-linked to antioxidant enzymes. TcCyp19 was found more concentrated in the membrane leading edges of the host cells in regions that also accumulate phosphorylated p47phox , as observed to the endogenous cyclophilin A, suggesting some mechanisms involved with the translocation process of the regulatory subunit p47phox in the activation of the NADPH oxidase enzymatic complex. We concluded that cyclophilin released in the host cell cytosol by T. cruzi mediates the increase of ROS, required to boost parasite proliferation in mammalian hosts.

EIF2α phosphorylation is regulated in intracellular amastigotes for the generation of infective Trypanosoma cruzi trypomastigote forms.

Trypanosomatids regulate gene expression mainly at the post-transcriptional level through processing, exporting and stabilising mRNA and control of translation. In most eukaryotes, protein synthesis is regulated by phosphorylation of eukaryotic initiation factor 2 (eIF2) at serine 51. Phosphorylation halts overall translation by decreasing availability of initiator tRNAmet to form translating ribosomes. In trypanosomatids, the N-terminus of eIF2α is extended with threonine 169 the homologous phosphorylated residue. Here, we evaluated whether eIF2α phosphorylation varies during the Trypanosoma cruzi life cycle, the etiological agent of Chagas' disease. Total levels of eIF2α are diminished in infective and non-replicative trypomastigotes compared with proliferative forms from the intestine of the insect vector or amastigotes from mammalian cells, consistent with decreased protein synthesis reported in infective forms. eIF2α phosphorylation increases in proliferative intracellular forms prior to differentiation into trypomastigotes. Parasites overexpressing eIF2αT169A or with an endogenous CRISPR/Cas9-generated eIF2αT169A mutation were created and analysis revealed alterations to the proteome, largely unrelated to the presence of µORF in epimastigotes. eIF2αT169A mutant parasites produced fewer trypomastigotes with lower infectivity than wild type, with increased levels of sialylated mucins and oligomannose glycoproteins, and decreased galactofuranose epitopes and the surface protease GP63 on the cell surface. We conclude that eIF2α expression and phosphorylation levels affect proteins relevant for intracellular progression of T. cruzi.

GCN2-Like Kinase Modulates Stress Granule Formation During Nutritional Stress in Trypanosoma cruzi.

The integrated stress response in eukaryotic cells is an orchestrated pathway that leads to eukaryotic Initiation Factor 2 alpha subunit (eIF2α) phosphorylation at ser51 and ultimately activates pathways to mitigate cellular damages. Three putative kinases (Tck1, Tck2, and Tck3) are found in the Trypanosoma cruzi genome, the flagellated parasite that causes Chagas disease. These kinases present similarities to other eukaryotic eIF2α kinases, exhibiting a typical insertion loop in the kinase domain of the protein. We found that this insertion loop is conserved among kinase 1 of several T. cruzi strains but differs among various Kinetoplastidae species, suggesting unique roles. Kinase 1 is orthologous of GCN2 of several eukaryotes, which have been implicated in the eIF2α ser51 phosphorylation in situations that mainly affects the nutrients levels. Therefore, we further investigated the responses to nutritional stress of T. cruzi devoid of TcK1 generated by CRISPR/Cas9 gene replacement. In nutrient-rich conditions, replicative T. cruzi epimastigotes depleted of TcK1 proliferate as wild type cells but showed increased levels of polysomes relative to monosomes. Upon nutritional deprivation, the polysomes decreased more than in TcK1 depleted line. However, eIF2α is still phosphorylated in TcK1 depleted line, as in wild type parasites. eIF2α phosphorylation increased at longer incubations times, but KO parasites showed less accumulation of ribonucleoprotein granules containing ATP-dependent RNA helicase involved in mRNA turnover (DHH1) and Poly-A binding protein (PABP1). Additionally, the formation of metacyclic-trypomastigotes is increased in the absence of Tck1 compared to controls. These metacyclics, as well as tissue culture trypomastigotes derived from the TcK1 knockout line, were less infective to mammalian host cells, although replicated faster inside mammalian cells. These results indicate that GCN2-like kinase in T. cruzi affects stress granule formation, independently of eIF2α phosphorylation upon nutrient deprivation. It also modulates the fate of the parasites during differentiation, invasion, and intracellular proliferation.

Diesel exhaust exposure intensifies inflammatory and structural changes associated with lung aging in mice.

Life expectancy is increasing worldwide. Lung aging is a process marked by changes in multiple morphological, physiological and age-related biomarkers (e.g., sirtuins) and is influenced by external factors, such as air pollution. Hence, the elderly are considered more vulnerable to the air pollution hazards. We hypothesized that diesel exhaust (DE) exposure intensifies changes in lung inflammatory and structural parameters in aging subjects. Two- and fifteen-month-old mice were exposed to DE for 30 days. Lung function was measured using the forced oscillation method. The inflammatory profile was evaluated in the bronchoalveolar lavage fluid (BALF) and blood, and lung volumes were estimated by stereology. Antioxidant enzyme activity was evaluated by spectrophotometry, sirtuin 1 (SIRT1), sirtuin 2 (SIRT2) and sirtuin 6 (SIRT6) expression was assessed by reverse transcription polymerase chain reaction (RT-PCR), and levels of the sirtuin proteins were evaluated by immunohistochemical staining in lung tissues. Older mice presented decreased pulmonary resistance and elastance, increased macrophage infiltration and decreased tumor necrosis factor (TNF) and interleukin 10 (IL-10) levels in the BALF, reduced activities of the antioxidant enzymes glutathione peroxidase (GPx) and glutathione reductase (GR), and increased activity glutathione S-transferase (GST); increased lung volumes with decreased elastic fiber and increased airway collagen content. SIRT1 gene expression was decreased in older animals, but protein levels were increased. DE exposure increased macrophage infiltration and oxidative stress in the lungs of animals of both ages. SIRT6 gene expression was decreased by DE exposure, with increased protein levels. In older animals, DE affected lung structure and collagen content. Lung aging features, such as decreased antioxidant reserves, lower IL-10 expression, and decreased SIRT1 levels may predispose subjects to exacerbated responses after DE exposure. Our data support the hypothesis that strategies designed to reduce ambient air pollution are an important step towards healthy aging.

Intervenção fonoaudiológica precoce em afasia infantil decorrente de um acidente vascular cerebral: relato de caso / Early speech-language intervention in childhood aphasia after a stroke: case report / Intervención fonoaudiológica precoz en afasia infantil debido a un accidente vascular cerebral: reporte de un caso

O Acidente Vascular Cerebral é menos frequente em crianças do que em adultos, porém possui um poder devastador para o futuro destes indivíduos, mesmo com indícios de melhor recuperação das alterações. O objetivo deste estudo é descrever os achados fonoaudiológicos de um caso de Acidente Vascular Cerebral infantil, evidenciando a evolução após 12 meses de terapia fonoaudiológica, iniciada precocemente, após lesão neurológica adquirida. Trata-se de um indivíduo de 11 anos, gênero masculino, com normalidade de desenvolvimento neuropsicomotor e independência para as atividades funcionais durante a primeira infância. Em março de 2015 sentiu um mal súbito, foi encaminhado para o hospital de urgência e diagnosticado com Acidente Vascular Cerebral, do tipo hemorrágico, em região frontoparietotemporal esquerda. Foram realizados procedimentos cirúrgicos, e a internação durou 25 dias. No momento da alta hospitalar houve a orientação sobre a necessidade de atendimento fonoaudiológico. A primeira avaliação de linguagem evidenciou afasia adquirida do tipo emissiva, conforme as classificações propostas para crianças. Foram realizadas 91 sessões de terapia fonoaudiológica, com duração de 50 minutos, pelo período de 12 meses. Ao analisar as habilidades formais de linguagem que permaneceram alteradas, observa-se que as relacionadas à leitura e escrita são as de maior significância, interferindo no desempenho escolar e desenvolvimento comunicativo. Assim, além do atendimento fonoaudiológico até que seja possível uma comunicação funcional, é importante que haja o apoio escolar.

Stroke is less common in children than in adults, but it has a devastating power for the future of these individuals, even with signs of better recovery from the changes. The aim of this study is to describe the speech therapy findings of a case of stroke child, showing the progress after 12 months of speech therapy initiated early after acquired neurological injury. This is an individual of 11 years old, male, with neurodevelopment normality and independence to the functional activities during early childhood. In March of 2015, he suffered a sudden illness, was referred to the emergency hospital and diagnosed with hemorrhagic stroke, in the left frontoparetotemporal region. Surgical procedures were performed, and hospitalization lasted 25 days. At the time of hospital discharge, there was guidance about the need for speech therapy care. In the first evaluation of language, there was evidence of acquired aphasia of the emissive type, according to the classifications proposed for children. A total of 91 speech therapy sessions lasting 50 minutes were performed for 12 months. When analyzing formal language skills that have remained altered, it is observed that those related to reading and writing are those of greater significance, interfering in school performance and communicative development. Thus, in addition to speech and language therapy until functional communication is possible, it is important that there be school support.

El accidente cerebrovascular es menos común en niños que en adultos, pero tiene un poder devastador para el futuro de estes individuos aun que se tenga indicios de mejor recuperación de las alteraciones. El objetivo de este estudio es describir los hallazgos fonoauiológicos de un caso de Accidente Vascular Cerebral Infantil, que muestra el progreso después de 12 meses de terapia fonoaudiológica iniciada precozmente, en un caso de lesión neurológica adquirida. Se trata de un individuo de 11 años, de sexo masculino, con normalidad el desarrollo neuropsicomotor y independencia de las actividades funcionales durante la primera infancia. En marzo el año 2015 sintió un mal súbito, fue remitido al hospital de emergencia y diagnosticado como Accidente Vascular Cerebral, de tipo hemorrágico en la región frontoparietotemporal izquierda. Se realizaron procedimientos quirúrgicos y la hospitalización duró 25 días. En el momento del alta hospitalaria hubo orientación sobre la necesidad de tratamiento fonoaudiológico. La primera evaluación del lenguaje mostró afasia adquirida de tipo emisivo, de acuerdo con las clasificaciones propuestas para los niños. Se realizaron 91 sesiones de terapia fonoaudiológica, con una duración de 50 minutos, el período de 12 meses. Mediante el análisis de las habilidades lingüísticas formales que permanecieron cambiada, se observa que las relacionadas con la lectura y la escritura son las más significativas, porque interfieren con el desempeño escolar y desarrollo comunicativo. Por lo tanto, además de la terapia fonoaudiologoca hasta que una comunicación funcional sea posible, es importante el apoyo escolar.

Polyphenol-Rich Diets Exacerbate AMPK-Mediated Autophagy, Decreasing Proliferation of Mosquito Midgut Microbiota, and Extending Vector Lifespan.

BACKGROUND: Mosquitoes feed on plant-derived fluids such as nectar and sap and are exposed to bioactive molecules found in this dietary source. However, the role of such molecules on mosquito vectorial capacity is unknown. Weather has been recognized as a major determinant of the spread of dengue, and plants under abiotic stress increase their production of polyphenols. RESULTS: Here, we show that including polyphenols in mosquito meals promoted the activation of AMP-dependent protein kinase (AMPK). AMPK positively regulated midgut autophagy leading to a decrease in bacterial proliferation and an increase in vector lifespan. Suppression of AMPK activity resulted in a 6-fold increase in midgut microbiota. Similarly, inhibition of polyphenol-induced autophagy induced an 8-fold increase in bacterial proliferation. Mosquitoes maintained on the polyphenol diet were readily infected by dengue virus. CONCLUSION: The present findings uncover a new direct route by which exacerbation of autophagy through activation of the AMPK pathway leads to a more efficient control of mosquito midgut microbiota and increases the average mosquito lifespan. Our results suggest for the first time that the polyphenol content and availability of the surrounding vegetation may increase the population of mosquitoes prone to infection with arboviruses.

Photo-assisted electrochemical degradation of simulated textile effluent coupled with simultaneous chlorine photolysis.

The influence of chloride ion concentration during the photo-assisted electrochemical degradation of simulated textile effluent, using a commercial Ti/Ru0.3Ti0.7O2 anode, was evaluated. Initially, the effect of applied current and supporting electrolyte concentration on the conversion of chloride ions to form reactive chlorine species in 90 min of experiment was analyzed in order to determine the maximum production of reactive chlorine species. The optimum conditions encountered (1.5 A and 0.3 mol dm(-3) NaCl) were subsequently employed for the degradation of simulated textile effluent. The efficiency of the process was determined through the analysis of chemical oxygen demand (COD), total organic carbon (TOC), of the presence of organochlorine products and phytotoxicity. Photo-assisted electrochemical degradation was more efficient for COD and TOC removal than the electrochemical technique alone. With simultaneous UV irradiation, a reduced quantity of reactive chlorine was produced, indicating that photolysis of the chlorine species led to the formation of hydroxyl radicals. This fact turns a simple electrochemical process into an advanced oxidation process.

Genome of Rhodnius prolixus, an insect vector of Chagas disease, reveals unique adaptations to hematophagy and parasite infection.

Rhodnius prolixus not only has served as a model organism for the study of insect physiology, but also is a major vector of Chagas disease, an illness that affects approximately seven million people worldwide. We sequenced the genome of R. prolixus, generated assembled sequences covering 95% of the genome (∼ 702 Mb), including 15,456 putative protein-coding genes, and completed comprehensive genomic analyses of this obligate blood-feeding insect. Although immune-deficiency (IMD)-mediated immune responses were observed, R. prolixus putatively lacks key components of the IMD pathway, suggesting a reorganization of the canonical immune signaling network. Although both Toll and IMD effectors controlled intestinal microbiota, neither affected Trypanosoma cruzi, the causal agent of Chagas disease, implying the existence of evasion or tolerance mechanisms. R. prolixus has experienced an extensive loss of selenoprotein genes, with its repertoire reduced to only two proteins, one of which is a selenocysteine-based glutathione peroxidase, the first found in insects. The genome contained actively transcribed, horizontally transferred genes from Wolbachia sp., which showed evidence of codon use evolution toward the insect use pattern. Comparative protein analyses revealed many lineage-specific expansions and putative gene absences in R. prolixus, including tandem expansions of genes related to chemoreception, feeding, and digestion that possibly contributed to the evolution of a blood-feeding lifestyle. The genome assembly and these associated analyses provide critical information on the physiology and evolution of this important vector species and should be instrumental for the development of innovative disease control methods.

Cyanoacrylate Adhesive as an Alternative Tool for Membrane Fixation in Guided Tissue Regeneration.

AIM: To report a well succeeded use of cyanoacrylate adhesive for fixating a resorbable membrane during a guided tissue regeneration procedure (GTR). BACKGROUND: The immobilization of membranes in GTR is essential for establishing proper environment for cell differentiation and tissue regeneration. However, some membranes are very difficult to be kept in position by sutures and its fixation by mini screws or pins may be time consuming and expensive. CASE DESCRIPTION: A 47-year-old woman presenting a vertical bone defect at the palatal aspect of the left central incisor was treated by GTR using particulate autogenous bone graft associated to a collagen membrane. The membrane was glued to the bone surrounding the defect and to the tooth surface with cyanoacrylate adhesive. The postoperative period was uneventful and 4 years later, excellent results in terms of radiographic filling of the defect and reduction of the probing depth were seen. For illustrative purposes, histological findings obtained during a previous experiment in calvaria of guinea pigs is shown, characterizing a foreign body granuloma and proving that the cyanoacrylate adhesive is a safe tool in GTR. CONCLUSION: The use of a membrane glued with cyanoacrylate to immobilize membranes in GTR is viable and safe from both technical and biological standpoints and may be advantageous for clinical and research purposes. CLINICAL SIGNIFICANCE: The alternative method for membrane fixation shown in this case report can contribute to simplify the technique in GTR procedures.

Proliferation and differentiation of Trypanosoma cruzi inside its vector have a new trigger: redox status.

Trypanosoma cruzi proliferate and differentiate inside different compartments of triatomines gut that is the first environment encountered by T. cruzi. Due to its complex life cycle, the parasite is constantly exposed to reactive oxygen species (ROS). We tested the influence of the pro-oxidant molecules H2O2 and the superoxide generator, Paraquat, as well as, metabolism products of the vector, with distinct redox status, in the proliferation and metacyclogenesis. These molecules are heme, hemozoin and urate. We also tested the antioxidants NAC and GSH. Heme induced the proliferation of epimastigotes and impaired the metacyclogenesis. ß-hematin, did not affect epimastigote proliferation but decreased parasite differentiation. Conversely, we show that urate, GSH and NAC dramatically impaired epimastigote proliferation and during metacyclogenesis, NAC and urate induced a significant increment of trypomastigotes and decreased the percentage of epimastigotes. We also quantified the parasite loads in the anterior and posterior midguts and in the rectum of the vector by qPCR. The treatment with the antioxidants increased the parasite loads in all midgut sections analyzed. In vivo, the group of vectors fed with reduced molecules showed an increment of trypomastigotes and decreased epimastigotes when analyzed by differential counting. Heme stimulated proliferation by increasing the cell number in the S and G2/M phases, whereas NAC arrested epimastigotes in G1 phase. NAC greatly increased the percentage of trypomastigotes. Taken together, these data show a shift in the triatomine gut microenvironment caused by the redox status may also influence T. cruzi biology inside the vector. In this scenario, oxidants act to turn on epimastigote proliferation while antioxidants seem to switch the cycle towards metacyclogenesis. This is a new insight that defines a key role for redox metabolism in governing the parasitic life cycle.

Avaliação periodontal e salivar de uma amostra de pacientes com Diabetes mellitus tipo 1 do município de Bauru-SP / Periodontal and salivary evaluation of a sample of type 1 diabetes mellitus patients of Bauru SP - Brazil

A inter-relação entre o diabetes mellitus (DM) e a doença periodontal (DP)aponta para o DM como fator de risco para DP, aumentando sua prevalência,extensão e severidade. Além disso, a DP é considerada a sexta complicação doDM. Diante da atual epidemia mundial de diabetes, incluindo aumento da incidênciade casos de diabetes mellitus tipo 1 (DM1) em pacientes cada vez mais jovens, oobjetivo deste trabalho observacional foi avaliar a condição periodontal e salivar deuma amostra de pacientes com DM1 da cidade de Bauru-SP. Trinta pacientesdivididos entre grupo teste (GDM1 – com diagnóstico de diabetes tipo 1) e grupocontrole (GC - normoglicêmicos) foram avaliados clinicamente através dosparâmetros periodontais - profundidade de sondagem (PS), nível clínico de inserção(NCI), índice de placa (IP) e índice de sangramento à sondagem (ISS) – e salivares– pH, capacidade tampão, glicose salivar e atividade da peroxidase, além dissotiveram a glicemia capilar mensurada. Para GDM1, foi solicitado a apresentação doresultado do teste de hemoglobina glicada (HbA1c) mais recente. A análiseestatística foi feita através do teste t de student para dados pareados e correlaçãode Pearson (p<0,05). Entre os parâmetros clínicos periodontais, houve diferençaestatisticamente significante entre os grupos apenas para IP e ISS (p<0,01). Quandoas variáveis foram analisadas correlacionando-se aos resultados da HbA1c, ascorrelações positivas foram encontradas com IP e ISS (p<0,01). Os níveis deglicemia capilar apresentaram diferença estatisticamente significante entre grupos(p<0,01) e correlações positivas foram encontradas apenas no GC, nos parâmetrosPS e NCI (p˂0,05). Para análises salivares, nenhuma diferença estatisticamente foiencontrada entre os grupos (p>0,05). Concluiu-se que os pacientes com DM1apresentaram pior condição periodontal, porém sem diferenças nos parâmetrossalivares.

Diabetes Mellitus is a risk factor for periodontal disease increasing itsprevalence, extension and severity. Periodontal disease is considered the sixthcomplication of diabetes. There is a global epidemic of diabetes, including anincrease of incidence of type 1 diabetes (T1D) in younger patients. Thus, the aim ofthis observational study was to evaluate the periodontal and salivary condition of asample of patients with T1D of Bauru – SP – Brazil. Thirty patients were divided intest group (T1DG – type 1 diabetes) and control (CG – normoglycemic). Periodontalclinical parameters were evaluated by means of probing depth (PD), bleeding onprobing (BOP), clinical attachment level (CAL), plaque index (PI). Salivary testsconsisted of pH, buffer capacity, salivary glucose and peroxidases activity. Thecapillary glycemia was recorded for all patients and the glycated hemoglobin (HbA1c)for the T1DG. The statistical analysis was performed through Student’s t test forcomparisons between groups and Pearson’s correlation (p<0.05). Results showedstatistically significant differences for groups for PI and BOP (p<0.01). Positivecorrelations were found for HbA1c, PI and BOP (p<0.01). Levels of capillary glycemiawere statistically significant differences between groups (p<0.01). Positivecorrelations between capillary glycemia, PD and NCI were encountered only forcontrol group (p<0.01). There was no significant difference between groups forsalivary analysis (p>0.05). In conclusion, T1D patients present worse periodontalconditions and no difference in salivary parameters.