Effect of genistein and coenzyme Q10 in oxidative damage and mitochondrial membrane potential in an attenuated type II mucopolysaccharidosis cellular model.

Mucopolysaccharidosis type II (MPS II) is an inborn error of the metabolism resulting from several possible mutations in the gene coding for iduronate-2-sulfatase (IDS), which leads to a great clinical heterogeneity presented by these patients. Many studies demonstrate the involvement of oxidative stress in the pathogenesis of inborn errors of metabolism, and mitochondrial dysfunction and oxidative stress can be related since most of reactive oxygen species come from mitochondria. Cellular models have been used to study different diseases and are useful in biochemical research to investigate them in a new promising way. The aim of this study is to develop a heterozygous cellular model for MPS II and analyze parameters of oxidative stress and mitochondrial dysfunction and investigate the in vitro effect of genistein and coenzyme Q10 on these parameters for a better understanding of the pathophysiology of this disease. The HP18 cells (heterozygous c.261_266del6/c.259_261del3) showed almost null results in the activity of the IDS enzyme and presented accumulation of glycosaminoglycans (GAGs), allowing the characterization of this knockout cellular model by MPS II gene editing. An increase in the production of reactive species was demonstrated (p < .05 compared with WT vehicle group) and genistein at concentrations of 25 and 50 µm decreased in vitro its production (p < .05 compared with HP18 vehicle group), but there was no effect of coenzyme Q10 in this parameter. There was a tendency for lysosomal pH change in HP18 cells in comparison to WT group and none of the antioxidants tested demonstrated any effect on this parameter. There was no increase in the activity of the antioxidant enzymes superoxide dismutase and catalase and oxidative damage to DNA in HP18 cells in comparison to WT group and neither genistein nor coenzyme q10 had any effect on these parameters. Regarding mitochondrial membrane potential, genistein induced mitochondrial depolarization in both concentrations tested (p < .05 compared with HP18 vehicle group and compared with WT vehicle group) and incubation with coenzyme Q10 demonstrated no effect on this parameter. In conclusion, it is hypothesized that our cellular model could be compared with a milder MPS II phenotype, given that the accumulation of GAGs in lysosomes is not as expressive as another cellular model for MPS II presented in the literature. Therefore, it is reasonable to expect that there is no mitochondrial depolarization and no DNA damage, since there is less lysosomal impairment, as well as less redox imbalance.

Increased cytokine levels induced by high phenylalanine concentrations in late diagnosis PKU patients compared to early diagnosis: Anti-inflammatory effect of L-carnitine.

Phenylketonuria (PKU) was the first genetic disease to have an effective therapy, which consists of phenylalanine intake restriction. However, there are patients who do not adhere to treatment and/or are not submitted to neonatal screening. PKU patients present L-carnitine (L-car) deficiency, compound that has demonstrated an antioxidant and anti-inflammatory role in metabolic diseases. This study evaluated the effect caused by exposure time to high Phe levels in PKU patients at early and late diagnosis, through pro- and anti-inflammatory cytokines, as well as the L-car effect in patients under treatment. It was observed that there was a decrease in phenylalanine levels in treated patients compared to patients at diagnosis, and an increase in L-car levels in the patients under treatment. Inverse correlation between Phe versus L-car and nitrate plus nitrite versus L-car in PKU patients was also showed. We found increased proinflammatory cytokines levels: interleukin (IL)-1ß, interferons (IFN)-gamma, IL-2, tumor necrosis factor (TNF)-alpha, IL-8 and IL-6 in the patients at late diagnosis compared to controls, and IL-8 in the patients at early diagnosis and treatment compared to controls. Increased IL-2, TNF-alpha, IL-6 levels in the patients at late diagnosis compared to early diagnosis were shown, and reduced IL-6 levels in the treated patients compared to patients at late diagnosis. Moreover, it verified a negative correlation between IFN-gamma and L-car in treated patients. Otherwise, it was observed that there were increased IL-4 levels in the patients at late diagnosis compared to early diagnosis, and reduction in treated patients compared to late diagnosed patients. In urine, there was an increase in 8-isoprostane levels in the patients at diagnosis compared to controls and a decrease in oxidized guanine species in the treated patients compared to the diagnosed patients. Our results demonstrate for the first time in literature that time exposure to high Phe concentrations generates a proinflammatory status, especially in PKU patients with late diagnosis. A pro-oxidant status was verified in not treated PKU patients. Our results demonstrate the importance of early diagnosis and prompt start of treatment, in addition to the importance of L-car supplementation, which can improve cellular defense against inflammation and oxidative damage in PKU patients.

Factors associated with women diagnosed with syphilis who received prenatal care in a primary healthcare unit.

OBJECTIVE: To estimate the prevalence of syphilis and its associated factors in women who were treated at public maternity hospitals and received prenatal care in a primary healthcare unit. METHODS: This cross-sectional study included 399 postpartum women. Interviews were conducted, and additional data were extracted from the pregnant woman's booklet, medical records, and printed tests. The dependent variable was a gestational syphilis diagnosis. The independent variables were grouped into socioeconomic and demographic, behavioral, reproductive, and prenatal blocks. The prevalence, prevalence ratios, and 95% confidence intervals (95%CI) were calculated. The χ 2 test was also performed (p≤0.05). Multivariate analysis was performed using Poisson regression models. RESULTS: The prevalence of gestational syphilis was 9.61% (95%CI: 7.14-12.83). We identified the following determining factors (adjusted prevalence ratios): history of sexually transmitted infections (2.3), first sexual intercourse by the age of 15 (2.42), partner having a history of syphilis (5.98), partner using crack/cocaine (6.42) and marijuana and others (3.02), not having a partner (3.07), low income (2.85), history of stillbirth (5.21), beginning prenatal care in the third trimester (3.15), and prenatal care received in a primary healthcare unit (without a Family Health Strategy team) (0.35). CONCLUSION: Individual and prenatal factors were associated with gestational syphilis. To control congenital syphilis, targeted interventions are needed to control syphilis in the adult population including expansion of access to quality prenatal care with identification of risks for syphilis and connection between prevention and treatment actions, implementation of strategies focused on early sexual education, effective establish prenatal care involving both partners, and effective implementation of the National Men's Health Policy (PNAISH - Política Nacional de Atenção Integral à Saúde dos Homens ).

Inflammatory process and oxidative/nitrative stress: in vivo study in mucopolysaccharidosis type IV A patients under long-term enzyme replacement therapy.

Mucopolysaccharidosis type IV A (MPS IVA) is an inborn error of the metabolism (IEM) caused by a deficiency of the enzyme N-acetylgalactosamine 6-sulfate sulfatase (GALNS). Since 2014, enzyme replacement therapy (ERT) is the recommended treatment for these patients. It is known that the inflammatory response is closely related to antioxidant defenses and oxidative stress, and literature shows involvement of oxidative stress in the pathogenesis of IEM. The aim of this study is to investigate the mechanisms of oxidative/nitrative stress and inflammation in patients with MPS IVA under long-term ERT. In the present work we investigate parameters of oxidative/nitrative stress in plasma and urine of MPS IVA patients under long-term ERT and controls, such as plasmatic nitrate/nitrite levels using the LDH Method, urinary di-tyrosine levels by fluorometric method, plasmatic content of sulfhydryl groups, urinary oxidized guanine species by ELISA kit and the plasmatic total antioxidant status. We next evaluated the plasmatic pro and anti-inflammatory cytokines concentration (IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α) and the expression of factors and enzymes Nrf-2, NF-κß and HO-1, main mediators between inflammation and oxidative stress. In concern to the oxidative/nitrative stress parameters, there was no significant difference between the groups MPS IVA patients under long-term ERT and controls, showing that there is no overproducing of RNS, no protein damage, no DNA/RNA oxidative damage and no modification in the non-enzymatic antioxidant capacity of a tissue to prevent the damage associated to free radical processes in these patients. It was also verified no significant difference between the MPS IVA patients under long-term ERT and controls groups regarding the production of proinflammatory cytokines. About anti-inflammatory cytokines, IL 10 was shown to be elevated in MPS IVA patients under long-term ERT in comparison to the control group. We next evaluated the genic expression of Nrf-2, NF-κß and HO-1and there was no significant difference between the MPS IVA patients under long-term ERT and control groups. In conclusion, MPS IVA patients under long term ERT are not in an inflammatory state and there is no alteration in genic expression in the genes analyzed which are involved in oxidative stress and inflammatory pathways. It is,however, important to consider that absence of imbalance of antioxidant defenses in MPS IVA patients under long term ERT is so far preliminary it is supported by methodologies that are not highly sensitive nor very accurate. Further experiments in future using state-of-the-art methodologies will corroborate these findings. Nevertheless, our results demonstrated the protective effect of the treatment in relation to the parameters studied and the importance of starting treatment in the early stages of the disease.

Evaluation of oxidative damage to biomolecules and inflammation in patients with urea cycle disorders.

Urea cycle disorders (UCD) are inborn errors of metabolism that occur due to a loss of function in enzymes and transporters involved in the urea cycle, causing an intoxication by hyperammonemia and accumulation of metabolites. Patients can develop hepatic encephalopathy (HE), severe neurological and motor disabilities, and often death. The mechanisms involved in the pathophysiology of UCD are many and complex, but there are strong indications that oxidative stress and inflammation are present, being responsible for at least part of the cellular damage that occurs in these diseases. The aim of this study was to evaluate oxidative and nitrosative damage and inflammation in UCD, to better understand the pathophysiology mechanisms of these diseases. We evaluated the nitrite and nitrate content, thiobarbituric acid-reactive substances (TBARS), carbonyl protein content and a panel of cytokines in plasma sample of 14 patients. The UCD patients group consisted of individuals affected with ornithine transcarbamylase deficiency (n = 8), carbamoyl phosphate synthetase deficiency (n = 2), argininosuccinate synthetase deficiency (n = 2); arginase 1 deficiency (n = 1) and argininosuccinate lyase deficiency (n = 1). Patients mean age at diagnosis was 5.25 ± 9.86 years-old and mean concentrations were compared with healthy individuals of matched age and gender. We found a significant reduction in nitrogen reactive species in patients when compared to controls. TBARS was increased in patients, indicating lipid peroxidation. To evaluate protein oxidative damage in UCD, the carbonyl content was measured, and the results also demonstrated an increase in this biomarker. Finally, we found that UCD patients have enhanced concentrations of cytokines, with pro-inflammatory interleukins IL-6, IL-8, interferon-γ and TNF-α, and anti-inflammatory IL-10 being increased when compared to the control group. In conclusion, our results demonstrate that oxidative stress and inflammation occurs in UCD and probably contribute to the severe brain damage present in patients.

Factors associated with women diagnosed with syphilis who received prenatal care in a primary healthcare unit

ABSTRACT Objective To estimate the prevalence of syphilis and its associated factors in women who were treated at public maternity hospitals and received prenatal care in a primary healthcare unit. Methods This cross-sectional study included 399 postpartum women. Interviews were conducted, and additional data were extracted from the pregnant woman's booklet, medical records, and printed tests. The dependent variable was a gestational syphilis diagnosis. The independent variables were grouped into socioeconomic and demographic, behavioral, reproductive, and prenatal blocks. The prevalence, prevalence ratios, and 95% confidence intervals (95%CI) were calculated. The χ 2 test was also performed (p≤0.05). Multivariate analysis was performed using Poisson regression models. Results The prevalence of gestational syphilis was 9.61% (95%CI: 7.14-12.83). We identified the following determining factors (adjusted prevalence ratios): history of sexually transmitted infections (2.3), first sexual intercourse by the age of 15 (2.42), partner having a history of syphilis (5.98), partner using crack/cocaine (6.42) and marijuana and others (3.02), not having a partner (3.07), low income (2.85), history of stillbirth (5.21), beginning prenatal care in the third trimester (3.15), and prenatal care received in a primary healthcare unit (without a Family Health Strategy team) (0.35). Conclusion Individual and prenatal factors were associated with gestational syphilis. To control congenital syphilis, targeted interventions are needed to control syphilis in the adult population including expansion of access to quality prenatal care with identification of risks for syphilis and connection between prevention and treatment actions, implementation of strategies focused on early sexual education, effective establish prenatal care involving both partners, and effective implementation of the National Men's Health Policy (PNAISH - Política Nacional de Atenção Integral à Saúde dos Homens ).

Neural cell adhesion molecule is required for ventricular conduction system development.

The most distal portion of the ventricular conduction system (VCS) contains cardiac Purkinje cells (PCs), which are essential for synchronous activation of the ventricular myocardium. Contactin-2 (CNTN2), a member of the immunoglobulin superfamily of cell adhesion molecules (IgSF-CAMs), was previously identified as a marker of the VCS. Through differential transcriptional profiling, we discovered two additional highly enriched IgSF-CAMs in the VCS: NCAM-1 and ALCAM. Immunofluorescence staining showed dynamic expression patterns for each IgSF-CAM during embryonic and early postnatal stages, but ultimately all three proteins became highly enriched in mature PCs. Mice deficient in NCAM-1, but not CNTN2 or ALCAM, exhibited defects in PC gene expression and VCS patterning, as well as cardiac conduction disease. Moreover, using ST8sia2 and ST8sia4 knockout mice, we show that inhibition of post-translational modification of NCAM-1 by polysialic acid leads to disrupted trafficking of sarcolemmal intercalated disc proteins to junctional membranes and abnormal expansion of the extracellular space between apposing PCs. Taken together, our data provide insights into the complex developmental biology of the ventricular conduction system.

Treating COVID-19 With Hydroxychloroquine (TEACH): A Multicenter, Double-Blind Randomized Controlled Trial in Hospitalized Patients.

BACKGROUND: Effective therapies to combat coronavirus 2019 (COVID-19) are urgently needed. Hydroxychloroquine (HCQ) has in vitro antiviral activity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but the clinical benefit of HCQ in treating COVID-19 is unclear. Randomized controlled trials are needed to determine the safety and efficacy of HCQ for the treatment of hospitalized patients with COVID-19. METHODS: We conducted a multicenter, double-blind randomized clinical trial of HCQ among patients hospitalized with laboratory-confirmed COVID-19. Subjects were randomized in a 1:1 ratio to HCQ or placebo for 5 days and followed for 30 days. The primary efficacy outcome was a severe disease progression composite end point (death, intensive care unit admission, mechanical ventilation, extracorporeal membrane oxygenation, and/or vasopressor use) at day 14. RESULTS: A total of 128 patients were included in the intention-to-treat analysis. Baseline demographic, clinical, and laboratory characteristics were similar between the HCQ (n = 67) and placebo (n = 61) arms. At day 14, 11 (16.4%) subjects assigned to HCQ and 6 (9.8%) subjects assigned to placebo met the severe disease progression end point, but this did not achieve statistical significance (P = .350). There were no significant differences in COVID-19 clinical scores, number of oxygen-free days, SARS-CoV-2 clearance, or adverse events between HCQ and placebo. HCQ was associated with a slight increase in mean corrected QT interval, an increased D-dimer, and a trend toward an increased length of stay. CONCLUSIONS: In hospitalized patients with COVID-19, our data suggest that HCQ does not prevent severe outcomes or improve clinical scores. However, our conclusions are limited by a relatively small sample size, and larger randomized controlled trials or pooled analyses are needed.

Using online databases to produce comprehensive accounts of the vascular plants from the Brazilian protected areas: The Parque Nacional do Itatiaia as a case study.

BACKGROUND: Brazil is one of the most biodiverse countries in the world, with about 37,000 species of land plants. Part of this biodiversity is within protected areas. The development of online databases in the last years greatly improved the available biodiversity data. However, the existing databases do not provide information about the protected areas in which individual plant species occur. The lack of such information is a crucial gap for conservation actions. This study aimed to show how the information captured from online databases, cleaned by a protocol and verified by taxonomists allowed us to obtain a comprehensive list of the vascular plant species from the "Parque Nacional do Itatiaia", the first national park founded in Brazil. All existing records in the online database JABOT (15,100 vouchers) were downloaded, resulting in 11,783 vouchers identified at the species level. Overall, we documented 2,316 species belonging to 176 families and 837 genera of vascular plants in the "Parque Nacional do Itatiaia". Considering the whole vascular flora, 2,238 species are native and 78 are non-native. NEW INFORMATION: The "Parque Nacional do Itatiaia" houses 13% of the angiosperm and 37% of the fern species known from the Brazilian Atlantic Forest. Amongst these species, 82 have been cited as threatened, following IUCN categories (CR, EN or VU), seven are data deficient (DD) and 15 have been classified as a conservation priority, because they are only known from a single specimen collected before 1969.

A list of land plants of Parque Nacional do Caparaó, Brazil, highlights the presence of sampling gaps within this protected area.

BACKGROUND: Brazilian protected areas are essential for plant conservation in the Atlantic Forest domain, one of the 36 global biodiversity hotspots. A major challenge for improving conservation actions is to know the plant richness, protected by these areas. Online databases offer an accessible way to build plant species lists and to provide relevant information about biodiversity. A list of land plants of "Parque Nacional do Caparaó" (PNC) was previously built using online databases and published on the website "Catálogo de Plantas das Unidades de Conservação do Brasil." Here, we provide and discuss additional information about plant species richness, endemism and conservation in the PNC that could not be included in the List. We documented 1,791 species of land plants as occurring in PNC, of which 63 are cited as threatened (CR, EN or VU) by the Brazilian National Red List, seven as data deficient (DD) and five as priorities for conservation. Fifity-one species were possible new ocurrences for ES and MG states. NEW INFORMATION: "Parque Nacional do Caparaó" houses 8% of the land plant species endemic to the Brazilian Atlantic Forest, including 6% of its angiosperms, 31% of its lycophytes and ferns and 14% of its avascular plants. Twelve percent of the threatened species listed for the State of Espírito Santo and 7% listed for the State of Minas Gerais are also protected by PNC. Surprisingly, 79% of the collections analysed here were carried out in Minas Gerais, which represents just 21% of the total extension of the Park. The compiled data uncover a huge botanical collection gap in this federally-protected area.

Expansión rápida del maxilar superior y su impacto a nivel de la vía aérea superior: revisión bibliográfica / Rapid expansion of the maxilla and its impact on the upper airway aspect: bibliographic review

La expansión del maxilar ya sea rápida o lenta, ha sido un procedimiento utilizado duran- te años para la corrección de maloclusiones transversales. Sus beneficios y efectos a nivel dentario, como de sutura palatina media son conocidos. Sin embargo, en los últimos años la repercusión y sus efectos a nivel de vía aérea han sido estudiados y se han estable- cido como un punto de interés sobre todo durante el tratamiento de pacientes con pro- blemas respiratorios. Según la bibliografía actual, la expansión rápida del maxilar induce un aumento significativo del volumen de la vía aérea superior. Hoy en día, con el alto desarrollo de técnicas basadas en imágenes 3D y con el advenimiento de la tomografía computarizada Cone Beam, es posible realizar análisis cuantitativos y evaluar los efectos de la expansión maxilar a nivel de la vía aérea superior en los tres sentidos del espacio. Mediante el uso de imagenología es posible visualizar la ganancia en la vía aérea superior pero aún no es posible evidenciar si este aumento se correlaciona con una mejora en la función respiratoria

Maxillary expansion, whether rapid or slow, has been a procedure used for years to co- rrect transverse malocclusions. Its benefits and effects at the dental aspect, such as the middle palatal suture are well known. Lately, however, its repercussion and effects at the airway aspect, have been studied and established as a point of interest; especially during the treatment of patients with respiratory problems. According to current bibliography, rapid maxillary expansion is associated with an increase in the upper airway volume. Nowadays, with the high development of techniques based on 3D images and with the inception of Cone Beam computed tomography, it is possible to perform quantitative analysis and evaluate the effects of maxillary expansion on the upper airway in a three dimensional way. With the use of imagenology, it is possible to visualize the increase in the upper airway dimensions, however, there is no evidence that this increase somehow, improves the respiratory function

Prevention by L-carnitine of DNA damage induced by 3-hydroxy-3-methylglutaric and 3-methylglutaric acids and experimental evidence of lipid and DNA damage in patients with 3-hydroxy-3-methylglutaric aciduria.

3-hydroxy-3-methylglutaric aciduria (HMGA) is an inherited disorder of the leucine catabolic pathway in which occurs a deficiency of the 3-hydroxy-3-methylglutaryl-CoA lyase enzyme. Therefore, the organic acids 3-hydroxy-3-methylglutaric (HMG) and 3-methylglutaric (MGA), mainly, accumulate in tissues of affected patients. Lately, much attention has been focused on free radicals as mediators of tissue damage in human diseases, causing lipid peroxidation, protein oxidation and DNA damage. The treatment of this disease is based in a restricted protein ingest and supplementation with l-carnitine (LC), an antioxidant and detoxifying agent. In the present work, we investigated the in vitro oxidative damage to DNA induced by the accumulation of organic acids and oxidative stress parameters in vivo of patients with 3-HMG, as well as the effect of the recommended therapy. The in vitro DNA damage was analyzed by the alkaline comet assay in leukocytes incubated with HMG and MGA (1 mM, 2.5 mM and 5 mM) and co-incubated with LC (90 µM and 150 µM). The in vivo urinary 15-F2t-isoprostane levels and urinary oxidized guanine species were measured by ELISA kits in patient's urine before and after the treatment with LC. HMG and MGA induced a DNA damage index (DI) significantly higher than that of the control group. The DI was significantly reduced in the presence of LC. It was also verified a significant increase of oxidized guanine species and urinary isoprostane levels, biomarker of oxidative DNA damage and lipid peroxidation respectively, in patients before treatment. After the treatment and supplementation with LC, patients presented significantly lower levels of those biomarkers. Analyzing the data together, we can conclude that HMGA patients present oxidative lipid and DNA damage, which is induced by HMG and MGA, and the antioxidant therapy with LC can prevent that kind of injuries.

Outer-membrane protein LptD (PA0595) plays a role in the regulation of alginate synthesis in Pseudomonas aeruginosa.

PURPOSE: The presence of alginate-overproducing (Alg+) strains of Pseudomonas aeruginosa in cystic fibrosis patients is indicative of chronic infection. The Alg+ phenotype is generally due to a mutation in the mucA gene, encoding an innermembrane protein that sequesters AlgT/U, the alginate-specific sigma factor. AlgT/U release from the anti-sigma factor MucA is orchestrated via a complex cascade called regulated intramembrane proteolysis. The goal of this study is to identify new players involved in the regulation of alginate production. METHODOLOGY: Previously, a mutant with a second-site suppressor of alginate production (sap), sap27, was isolated from the constitutively Alg+ PDO300 that harbours the mucA22 allele. A cosmid from a P. aeruginosa minimum tiling path library was identified via en masse complementation of sap27. The cosmid was transposon mutagenized to map the contributing gene involved in the alginate production. The identified gene was sequenced in sap27 along with algT/U, mucA, algO and mucP. The role of the novel gene was explored using precise in-frame algO and algW deletion mutants of PAO1 and PDO300.Results/Key findings. The gene responsible for restoring the mucoid phenotype was mapped to lptD encoding an outer-membrane protein. However, the sequencing of sap27 revealed a mutation in algO, but not in lptD. In addition, we demonstrate that lipopolysaccharide transport protein D (LptD)-dependent alginate production requires AlgW in PAO1 and AlgO in PDO300. CONCLUSION: LptD plays a specific role in alginate production. Our findings suggest that there are two pathways for the production of alginate in P. aeruginosa, one involving AlgW in the wild-type, and one involving AlgO in the mucA22 mutant.

Pseudomonas aeruginosa Regulated Intramembrane Proteolysis: Protease MucP Can Overcome Mutations in the AlgO Periplasmic Protease To Restore Alginate Production in Nonmucoid Revertants.

The progression of cystic fibrosis (CF) from an acute to a chronic disease is often associated with the conversion of the opportunistic pathogen Pseudomonas aeruginosa from a nonmucoid form to a mucoid form in the lung. This conversion involves the constitutive synthesis of the exopolysaccharide alginate, whose production is under the control of the AlgT/U sigma factor. This factor is regulated posttranslationally by an extremely unstable process and has been commonly attributed to mutations in the algT (algU) gene. By exploiting this unstable phenotype, we isolated 34 spontaneous nonmucoid variants arising from the mucoid strain PDO300, a PAO1 derivative containing the mucA22 allele commonly found in mucoid CF isolates. Complementation analysis using a minimal tiling path cosmid library revealed that most of these mutants mapped to two protease-encoding genes, algO, also known as prc or PA3257, and mucP Interestingly, our algO mutations were complemented by both mucP and algO, leading us to delete, clone, and overexpress mucP, algO, mucE, and mucD in both wild-type PAO1 and PDO300 backgrounds to better understand the regulation of this complex regulatory mechanism. Our findings suggest that the regulatory proteases follow two pathways for regulated intramembrane proteolysis (RIP), where both the AlgO/MucP pathway and MucE/AlgW pathway are required in the wild-type strain but where the AlgO/MucP pathway can bypass the MucE/AlgW pathway in mucoid strains with membrane-associated forms of MucA with shortened C termini, such as the MucA22 variant. This work gives us a better understanding of how alginate production is regulated in the clinically important mucoid variants of Pseudomonas aeruginosaIMPORTANCE Infection by the opportunistic pathogen Pseudomonas aeruginosa is the leading cause of morbidity and mortality seen in CF patients. Poor patient prognosis correlates with the genotypic and phenotypic change of the bacteria from a typical nonmucoid to a mucoid form in the CF lung, characterized by the overproduction of alginate. The expression of this exopolysaccharide is under the control an alternate sigma factor, AlgT/U, that is regulated posttranslationally by a series of proteases. A better understanding of this regulatory phenomenon will help in the development of therapies targeting alginate production, ultimately leading to an increase in the length and quality of life for those suffering from CF.

Oral health literacy: a concise review

According to the World Health Organization, Health Literacy (HL) corresponds to the cognitive and social abilities that determine the motivation and ability of individuals to gain access, to understand, and to use information in ways that provide and maintain good health. The development of HL in dentistry came late, and only in the last decade did it reach a level similar to that in the medical area. In dentistry, HL centered on the concept of Oral Health Literacy (OHL), defined as the degree to which individuals have the capacity to obtain, process, and understand basic information on oral health and services that is necessary to make appropriate health decisions. The evidence suggests that people with low HL have worse health status and greater use of medical resources, which results in an increase in costs in the general population. Determinants of the level of OHL include age, level of education, and socioeconomic level. These determinants are reflected in low oral health and in less access to information or less understanding regarding care, pathologies, or dental treatments. The instruments for measuring HL and OHL are mainly aimed at recognizing arithmetic and reading skills, which are not fully related to the ability of the people surveyed to find, understand and use information related to health. OHL is an important issue at the level of health programs, because knowledge of OHL helps in medical practice, in disease prevention and in health promotion. OHL instruments must have validated and demonstrate adequate psychometric properties.

Mindfulness como estratégia para o tratamento da obesidade infantil / Mindfulness as a strategy for the treatment of childhood obesity

Introdução: A obesidade infantil é um grave problema global de saúde pública. O manejo no tratamento dessa enfermidade é complexo, pois depende de adequação à mudança de hábitos da própria família. A técnica mindfulness pode ser eficaz nesse contexto pois atua tanto no ganho de peso quanto nos sintomas ansiosos. Objetivo: avaliar a prática mindfulness em crianças e adolescentes. Avaliar sua aplicabilidade aos pais no tratamento da obesidade infantil. Material e Métodos: ensaio clínico longitudinal com 28 pacientes atendidos no ambulatório especializado de hospital terciário, no período de junho de 2016 a junho de 2018. As atividades consistiram de cinco sessões mensais de grupos operativos de meditações guiadas e atividades semanais domiciliares programadas. Foram avaliados peso, altura, Índice de Massa Corporal (IMC), escore Z do IMC e circunferências da cintura, do abdome, do quadril e do braço antes de cada sessão mensal. Resultados: a média da idade dos participantes foi 10,7 ± 3,68 anos; o escore Z do IMC 3,13 ± 0,89, a circunferência abdominal 90,9 ± 15,86 cm. A média de IMC referente à primeira avaliação foi 29,1 ± 4,63 kg/m2 e 29,9 ± 3,81kg/m2 na última. Os dados de evolução de IMC, circunferência da cintura, circunferência abdominal, circunferência do quadril e circunferência do braço direito não apresentaram diferença significante entre os valores iniciais e finais. Entretanto, houve melhora no vínculo entre as crianças e seus responsáveis com algumas mudanças no comportamento familiar. Conclusão: A prática mindfulness em crianças e adolescentes, bem como em seus pais, não mostrou impacto na redução do peso dos pacientes avaliados. Entretanto, poderá, em longo prazo, levar a resultados positivos na qualidade de vida e na melhora dos hábitos alimentares, podendo levar a redução do peso.

Introduction: Childhood obesity is a serious global public health problem. Management of this disease is complex, as it depends on changes of family habits. The mindfulness technique can be effective in this context by acting both on weight gain and on anxious symptoms. Objective: to evaluate the practice of mindfulness in obese children and adolescents as well as its applicability to their parents. Material and Methods: Longitudinal clinical trial with 28 patients attended at a tertiary pediatric endocrinology outpatient clinic from June 2016 to June 2018, who participated to five monthly group mindfulness guided sessions and weekly programmed home activities. Weight, height, Body Mass Index and waist, abdomen, hip and arm circumference were evaluated before each monthly session. Results: mean chronological age of participants was 10.7 ± 3.68 years; Z score of BMI 3.13 ± 0.89, the abdominal circumference 90.9 ± 15.86 cm. Mean BMI on the first evaluation was 29.1 ± 4.63 kg / m2 and 29.9 ± 3.81 kg / m2 on last evaluation. Data on BMI evolution, waist circumference, waist circumference, hip circumference and right arm circumference did not present significant difference between the initial and final values. However, there has been an improvement in the relationship between the children and their caregivers with some changes in family behavior. Conclusion: mindfulness practice in children and adolescents, as well as in their parents, showed no impact on weight reduction of assessed patients. In the long run, this approach may lead to positive results in quality of life and improvement in eating habits, and therefore may lead to weight reduction.

Isolation and characterization of embryonic stem cell-derived cardiac Purkinje cells.

The cardiac Purkinje fiber network is composed of highly specialized cardiomyocytes responsible for the synchronous excitation and contraction of the ventricles. Computational modeling, experimental animal studies, and intracardiac electrical recordings from patients with heritable and acquired forms of heart disease suggest that Purkinje cells (PCs) may also serve as critical triggers of life-threatening arrhythmias. Nonetheless, owing to the difficulty in isolating and studying this rare population of cells, the precise role of PC in arrhythmogenesis and the underlying molecular mechanisms responsible for their proarrhythmic behavior are not fully characterized. Conceptually, a stem cell-based model system might facilitate studies of PC-dependent arrhythmia mechanisms and serve as a platform to test novel therapeutics. Here, we describe the generation of murine embryonic stem cells (ESC) harboring pan-cardiomyocyte and PC-specific reporter genes. We demonstrate that the dual reporter gene strategy may be used to identify and isolate the rare ESC-derived PC (ESC-PC) from a mixed population of cardiogenic cells. ESC-PC display transcriptional signatures and functional properties, including action potentials, intracellular calcium cycling, and chronotropic behavior comparable to endogenous PC. Our results suggest that stem-cell derived PC are a feasible new platform for studies of developmental biology, disease pathogenesis, and screening for novel antiarrhythmic therapies.

The potential therapeutic effect of guanosine after cortical focal ischemia in rats.

BACKGROUND AND PURPOSE: Stroke is a devastating disease. Both excitotoxicity and oxidative stress play important roles in ischemic brain injury, along with harmful impacts on ischemic cerebral tissue. As guanosine plays an important neuroprotective role in the central nervous system, the purpose of this study was to evaluate the neuroprotective effects of guanosine and putative cerebral events following the onset of permanent focal cerebral ischemia. METHODS: Permanent focal cerebral ischemia was induced in rats by thermocoagulation. Guanosine was administered immediately, 1 h, 3 h and 6 h after surgery. Behavioral performance was evaluated by cylinder testing for a period of 15 days after surgery. Brain oxidative stress parameters, including levels of ROS/RNS, lipid peroxidation, antioxidant non-enzymatic levels (GSH, vitamin C) and enzymatic parameters (SOD expression and activity and CAT activity), as well as glutamatergic parameters (EAAC1, GLAST and GLT1, glutamine synthetase) were analyzed. RESULTS: After 24 h, ischemic injury resulted in impaired function of the forelimb, caused brain infarct and increased lipid peroxidation. Treatment with guanosine restored these parameters. Oxidative stress markers were affected by ischemic insult, demonstrated by increased ROS/RNS levels, increased SOD expression with reduced SOD activity and decreased non-enzymatic (GSH and vitamin C) antioxidant defenses. Guanosine prevented increased ROS/RNS levels, decreased SOD activity, further increased SOD expression, increased CAT activity and restored vitamin C levels. Ischemia also affected glutamatergic parameters, illustrated by increased EAAC1 levels and decreased GLT1 levels; guanosine reversed the decreased GLT1 levels and did not affect the EAAC1 levels. CONCLUSION: The effects of brain ischemia were strongly attenuated by guanosine administration. The cellular mechanisms involved in redox and glutamatergic homeostasis, which were both affected by the ischemic insult, were also modulated by guanosine. These observations reveal that guanosine may represent a potential therapeutic agent in cerebral ischemia by preventing oxidative stress and excitotoxicity.

Neuropsychological performance as endophenotypes in extended schizophrenia families from the Central Valley of Costa Rica.

OBJECTIVE: The understanding of complex heritable psychiatric disorders such as schizophrenia could be clarified by examining endophenotypes within genetically isolated populations, such as the one found in the Central Valley of Costa Rica. The reduction of familial variability within a sample could allow the relationship between the cognitive and symptomatic manifestations of the illness and the genetic underpinnings to become more observable. This study investigates the neuropsychological test performances of 41 family members from four extended multiplex families within the Spanish origin population of the Central Valley of Costa Rica as potential endophenotypes for genetic studies. METHODS: Individuals with a diagnosis of schizophrenia or schizoaffective disorder were compared with unaffected relatives and 15 unrelated controls with no family history of schizophrenia. RESULTS: Although the sample size is small, the results confirm previous reports in the literature of deficits in working memory, executive function, processing speed, and verbal fluency in individuals with schizophrenia compared with controls and intermediate performance in nonpsychotic family members compared with controls. We also found several suggestive quantitative cognitive trait loci with log of the odds greater than 1.75. CONCLUSION: These findings suggest that the cognitive deficits in schizophrenia are consistent aspects of the illness, although their usefulness as endophenotypes for genetic studies remains unclear.