Preclinical evaluation of combined therapy with amiodarone and low-dose benznidazole in a mouse model of chronic Trypanosoma cruzi infection.

Chagasic chronic cardiomyopathy (CCC) is the primary clinical manifestation of Chagas disease (CD), caused by Trypanosoma cruzi. Current therapeutic options for CD are limited to benznidazole (Bz) and nifurtimox. Amiodarone (AMD) has emerged as most effective drug for treating the arrhythmic form of CCC. To address the effects of Bz and AMD we used a preclinical model of CCC. Female C57BL/6 mice were infected with T. cruzi and subjected to oral treatment for 30 consecutive days, either as monotherapy or in combination. AMD in monotherapy decreased the prolonged QTc interval, the incidence of atrioventricular conduction disorders and cardiac hypertrophy. However, AMD monotherapy did not impact parasitemia, parasite load, TNF concentration and production of reactive oxygen species (ROS) in cardiac tissue. Alike Bz therapy, the combination of Bz and AMD (Bz/AMD), improved cardiac electric abnormalities detected T. cruzi-infected mice such as decrease in heart rates, enlargement of PR and QTc intervals and increased incidence of atrioventricular block and sinus arrhythmia. Further, Bz/AMD therapy ameliorated the ventricular function and reduced parasite burden in the cardiac tissue and parasitemia to a degree comparable to Bz monotherapy. Importantly, Bz/AMD treatment efficiently reduced TNF concentration in the cardiac tissue and plasma and had beneficial effects on immunological abnormalities. Moreover, in the cardiac tissue Bz/AMD therapy reduced fibronectin and collagen deposition, mitochondrial damage and production of ROS, and improved sarcomeric and gap junction integrity. Our study underlines the potential of the Bz/AMD therapy, as we have shown that combination increased efficacy in the treatment of CCC.

Multi-Generation Ecosystem Selection of Rhizosphere Microbial Communities Associated with Plant Genotype and Biomass in Arabidopsis thaliana.

The role of the microbiome in shaping the host's phenotype has emerged as a critical area of investigation, with implications in ecology, evolution, and host health. The complex and dynamic interactions involving plants and their diverse rhizospheres' microbial communities are influenced by a multitude of factors, including but not limited to soil type, environment, and plant genotype. Understanding the impact of these factors on microbial community assembly is key to yielding host-specific and robust benefits for plants, yet it remains challenging. Here, we conducted an artificial ecosystem selection experiment for eight generations of Arabidopsis thaliana Ler and Cvi to select soil microbiomes associated with a higher or lower biomass of the host. This resulted in divergent microbial communities shaped by a complex interplay between random environmental variations, plant genotypes, and biomass selection pressures. In the initial phases of the experiment, the genotype and the biomass selection treatment had modest but significant impacts. Over time, the plant genotype and biomass treatments gained more influence, explaining ~40% of the variation in the microbial community's composition. Furthermore, a genotype-specific association of plant-growth-promoting rhizobacterial taxa, Labraceae with Ler and Rhizobiaceae with Cvi, was observed under selection for high biomass.

Benznidazole and amiodarone combined treatment attenuates cytoskeletal damage in Trypanosoma cruzi-infected cardiac cells.

Chagas disease (CD), a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi, is an important public health problem mainly in Latin America, leading to approximately 12,000 annual deaths. Current etiological treatment for CD is limited to two nitro compounds, benznidazole (Bz) and nifurtimox (Nif), both presenting relevant limitations. Different approaches have been employed to establish more effective and safer schemes to treat T. cruzi infection, mostly based on drug repurposing and combination therapies. Amiodarone (AMD), an antiarrhythmic medicament of choice for patients with the chronic cardiac form of CD, is also recognized as a trypanocidal agent. Therefore, our aim is to investigate the combined treatment Bz + AMD on trypomastigote viability, control of T. cruzi intracellular form proliferation, and recovery of the infection-induced cytoskeleton alterations in cardiac cells. The combination of Bz + AMD did not improve the direct trypanocidal effect of AMD on the infective blood trypomastigote and replicative intracellular forms of the parasite. Otherwise, the treatment of T. cruzi-infected cardiac cells with Bz plus AMD attenuated the infection-triggered cytoskeleton damage of host cells and the cytotoxic effects of AMD. Thus, the combined treatment Bz + AMD may favor parasite control and hamper tissue damage.

Experimental Combination Therapy with Amiodarone and Low-Dose Benznidazole in a Mouse Model of Trypanosoma cruzi Acute Infection.

Chagas disease (CD), caused by Trypanosoma cruzi, affects approximately 6 to 7 million people in Latin America, with cardiomyopathy being the clinical manifestation most commonly associated with patient death during the acute phase. The etiological treatment of CD is restricted to benznidazole (Bz) and nifurtimox (Nif), which involve long periods of administration, frequent side effects, and low efficacy in the chronic phase. Thus, combined therapies emerge as an important tool in the treatment of CD, allowing the reduction of Bz dose and treatment duration. In this sense, amiodarone (AMD), the most efficient antiarrhythmic drug currently available and prescribed to CD patients, is a potential candidate for combined treatment due to its known trypanocidal activity. However, the efficacy of AMD during the acute phase of CD and its interaction with Bz or Nif are still unknown. In the present study, using a well-established murine model of the acute phase of CD, we observed that the Bz/AMD combination was more effective in reducing the peak parasitemia than both monotherapy treatments. Additionally, the Bz/AMD combination reduced (i) interleukin-6 (IL-6) levels in cardiac tissue, (ii) P-wave duration, and (iii) frequency of arrhythmia in infected animals and (iv) restored gap junction integrity in cardiac tissue. Therefore, our study validates AMD as a promising candidate for combined therapy with Bz, reinforcing the strategy of combined therapy for CD. IMPORTANCE Chagas disease affects approximately 6 to 7 million people worldwide, with cardiomyopathy being the clinical manifestation that most commonly leads to patient death. The etiological treatment of Chagas disease is limited to drugs (benznidazole and nifurtimox) with relatively high toxicity and therapeutic failures. In this sense, amiodarone, the most effective currently available antiarrhythmic drug prescribed to patients with Chagas disease, is a potential candidate for combined treatment due to its known trypanocidal effect. In the present study, we show that combined treatment with benznidazole and amiodarone improves the trypanocidal effect and reduces cardiac damage in acutely T. cruzi-infected mice.

Treatment of a Post-biopsy Pulmonary Artery Pseudoaneurysm.

Peripheral pulmonary artery pseudoaneurysms (PPAP) following a lung biopsy are exceedingly rare but can lead to severe haemoptysis. Cases requiring treatment are usually managed using an endovascular approach. Nevertheless, successful percutaneous treatment has been described. Several embolic agents can be used to accomplish percutaneous embolization. We aim to report a successful percutaneous treatment of a post-biopsy PPAP using thrombin. Our patient developed a PPAP, with consequent large alveolar haemorrhage and haemoptysis, after a transthoracic lung biopsy. Because the patient had a mechanical mitral valve, she needed to be anticoagulated, impairing the spontaneous resolution of the pseudoaneurysm. Therefore, the PPAP had to be treated to safely anticoagulate the patient. A percutaneous approach was chosen given the very peripheric location of the PPAP. Treating PPAP with an endovascular approach can be challenging. With this report, we demonstrate that the percutaneous approach is a safe and efficient alternative method. We choose thrombin as an embolic agent given its safety profile and efficacy under anticoagulation. The good results of our intervention reinforce the applicability and efficacy of this kind of treatment approach to PPAP.

Characterization and trypanocidal activity of a ß-lapachone-containing drug carrier.

The treatment of Chagas disease (CD), a neglected parasitic condition caused by Trypanosoma cruzi, is still based on only two drugs, nifurtimox (Nif) and benznidazole (Bz), both of which have limited efficacy in the late chronic phase and induce severe side effects. This scenario justifies the continuous search for alternative drugs, and in this context, the natural naphthoquinone ß-lapachone (ß-Lap) and its derivatives have demonstrated important trypanocidal activities. Unfortunately, the decrease in trypanocidal activity in the blood, high toxicity to mammalian cells and low water solubility of ß-Lap limit its systemic administration and, consequently, clinical applications. For this reason, carriers as drug delivery systems can strategically maximize the therapeutic effects of this drug, overcoming the above mentioned restrictions. Accordingly, the aim of this study is to investigate the in vitro anti-T. cruzi effects of ß-Lap encapsulated in2-hydroxypropyl-ß-cyclodextrin (2HP-ß-CD) and its potential toxicity to mammalian cells.

Análise longitudinal das condições de vida e saúde de idosos com histórico de câncer: Estudo Saúde, Bem-Estar e Envelhecimento / Longitudinal analysis of living conditions and health of elderly people with a history of cancer: Study Health, Well-Being and Aging

O uso de dados autorrelatados para estimar o câncer na população, bem como para o conhecimento de seus determinantes, tem se mostrado útil em estudos epidemiológicos. Em acréscimo, o uso de dados genéticos em pesquisas sobre câncer e envelhecimento tem se mostrado promissor. No entanto, são escassos os estudos longitudinais sobre envelhecimento que analisaram tanto a validade do autorrelato de câncer como a prevalência do histórico da doença, acrescido da análise das condições de vida e saúde, e da presença de variantes patogênicas associadas ao câncer hereditário. Esta tese apresenta-se sob a forma de dois manuscritos. O primeiro manuscrito verificou a validade do autorrelato do histórico de câncer e descreveu a prevalência da doença, antes e após o linkage, entre a base de dados do Estudo Saúde, Bem-Estar e Envelhecimento (SABE) com a base de dados do Registro de Câncer de Base Populacional de São Paulo (RCBP-SP). O segundo manuscrito analisou as mudanças nas condições de vida e saúde de idosos com histórico de câncer ao longo de cinco anos, identificou os fatores associados em dois momentos distintos, e descreveu a presença de mutações patogênicas associadas ao câncer hereditário. O câncer autorreferido apresentou acurácia de 98,5%, sensibilidade de 82,1% (IC5%:74,7-89,4), especificidade de 95,6% (IC55%:94,2-97,0), valor preditivo positivo de 59,6% (IC95%:50,6-68,6) e valor preditivo negativo de 98,6% (IC95%:97,8-99,3). A prevalência de câncer após o linkage foi 8,8% (IC95%:7,4-10,3) na linha de base e 12,8% (IC95%:10,5-15,0) no seguimento. Não foi observado mudanças nas características comportamentais e de saúde nos idosos com histórico de câncer durante o período estudado. Por outro lado, observaram-se diferenças nos fatores associados entre o primeiro e o segundo momento. Na linha de base, foram associados ao histórico da doença possuir serviço privado de saúde (OR: 1,56; IC95%: 1,11-2,20) e perda de apetite (OR: 2,50; IC95%: 1,28-4,88). No seguimento, permaneceu associado a perda de apetite (OR: 2,43; IC95%: 1,43-4,13), acrescido de depressão (OR: 1,80; IC95%:1,06-3,05) e cor de pele branca (OR: 1,65; IC95%: 1,07-2,54). Por sua vez, foram identificados 28 idosos com pelo menos uma das 14 mutações patogênicas associadas ao câncer hereditário detectadas pelo sequenciamento completo do genoma, mas nenhuma mutação mostrou-se compatível ao seu respectivo fenótipo. O câncer autorrelatado mostrou-se válido para o uso em estudos longitudinais sobre envelhecimento. Em cinco anos de seguimento, verificou-se aumento da prevalência de câncer e variações nos fatores associados ao histórico da doença. Tais achados são importantes para a formulação de ações de prevenção e cuidado personalizado, bem como para o planejamento dos serviços de saúde e do seu acesso.

The use of self-reported data to estimate cancer in the population, as well as for the knowledge of its determinants, has been shown to be useful in epidemiological studies. In addition, the use of genetic data in cancer and aging research has shown promise. However, there are few longitudinal studies on aging that analyzed both the validity of self-report of cancer and the prevalence of the history of the disease, plus the analysis of living and health conditions, and the presence of pathogenic variants associated with hereditary cancer. This thesis is presented in the form of two manuscripts. The first manuscript verified the validity of the self-report of cancer history and described the prevalence of the disease, before and after the linkage, between the Health, Well-Being and Aging Study (SABE) database with the Population-Based Cancer Registry of São Paulo (RCBP-SP). The second manuscript analyzed changes in the living and health conditions of elderly people with a history of cancer over a five-year period, identified the associated factors at two different times, and described the presence of pathogenic mutations associated with hereditary cancer. Self-reported cancer had an accuracy of 98.5%, sensitivity of 82.1% (95%CI: 74.7-89.4), specificity of 95.6% (95%CI: 94.2-97.0), positive predictive value of 59.6% (95%CI: 50.6-68.6) and negative predictive value of 98.6% (95%CI: 97.8-99.3). The prevalence of cancer after linkage was 8.8% (95%CI:7.4-10.3) at baseline and 12.8% (95%CI:10.5-15.0) at follow-up. No changes in behavioral and health characteristics were observed in the elderly with a history of cancer during the study period. On the other hand, differences were observed in the associated factors between the first and second moments. At baseline, having a private health service (OR: 1.56; 95%CI: 1.11-2.20) and loss of appetite (OR: 2.50; 95%CI: 1.28-4.88). During follow-up, it remained associated with loss of appetite (OR: 2.43; 95%CI: 1.43-4.13), plus depression (OR: 1.80; 95%CI: 1.06-3.05) and white skin color (OR: 1.65; 95%CI: 1.07-2.54). In turn, 28 elderly individuals were identified with at least one of the 14 pathogenic mutations associated with hereditary cancer detected by complete genome sequencing, but no mutation was compatible with their respective phenotype. Self-reported cancer has been shown to be valid for use in longitudinal studies on aging. In five years of follow-up, there was an increase in the prevalence of cancer and variations in factors associated with the history of the disease. These findings are important for the formulation of prevention and personalized care actions, as well as for the planning of health services and their access.

Development of auditory and language skills in children using cochlear implants with two signal processing strategies / Desenvolvimento das habilidades auditivas e de linguagem em crianças usuárias de implante coclear com duas estratégias de processamento de sinal

Abstract Introduction: The increase in the spectral information offered by the sound processing strategy HiRes 120 has led to great expectations for the pediatric population. Due to a shorter duration of auditory deprivation and higher neural plasticity, children could benefit more substantially from the spectral information of this sound processing strategy. Objective: To compare auditory and language skills in Brazilian children with cochlear implants using the HiRes and HiRes 120 sound processing strategies. Methods: Thirty children, aged 1-3 years, with congenital hearing loss, were divided into two groups, according to the signal processing strategy adjusted at the time of the cochlear implant activation. The assessed children were matched according to chronological age and the time of the cochlear implant use. The auditory and language skills were evaluated longitudinally through the Infant-Toddler Meaningful Auditory Integration Scale and Production Infant Scale Evaluation, carried out before surgery, and 3, 6 and 12 months after device implantation. The Mann-Whitney test was applied for the comparison between the two groups with a 5% significance level. Results: The findings indicated development of hearing and language skills in the first year of cochlear implant use; however, there was no statistically significant difference in the evolution of such skills due to the adjusted processing strategy in the activation of the cochlear implant electrodes. Conclusion: The development of auditory and language skills in the assessed children was similar during the entire study period, regardless of which signal processing strategy was used.

Resumo Introdução: O aumento das informações espectrais proporcionado pela estratégia de processamento de sinal HiRes 120 gera expectativas em relação à população pediátrica, uma vez que, devido ao curto período de privação sensorial e da maior plasticidade neural, as crianças poderiam se beneficiar de forma mais substancial das informações espectrais dessa estratégia de processamento de sinal. Objetivo: Comparar as habilidades auditivas e de linguagem em crianças usuárias de implante coclear, com a estratégia de processamento de sinal HiRes e HiRes 120. Método: Trinta crianças de um a três anos e com perda auditiva congênita foram divididas em dois grupos, de acordo com a estratégia de processamento de sinal ajustada no momento da ativação do implante coclear. As crianças avaliadas foram pareadas de acordo com a idade cronológica e o tempo de uso do implante coclear. As habilidades auditivas e de linguagem foram avaliadas, longitudinalmente, por meio do Infant-Toddler Meaningful Auditory Integration Scale e Production Infant Scale Evaluation, antes do implante coclear e com 3, 6 e 12 meses de uso do dispositivo. O teste estatístico de Mann-Whitney foi aplicado para comparar os resultados dos grupos, com nível de significância de 5%. Resultados: Os achados indicaram desenvolvimento das habilidades auditivas e de linguagem no primeiro ano de uso do implante coclear, porém sem diferença estatisticamente significante na evolução de tais habilidades, em função da estratégia de processamento ajustada na ativação dos eletrodos do implante coclear. Conclusão: O desenvolvimento das habilidades auditivas e de linguagem das crianças avaliadas foi semelhante durante o período do presente estudo, independentemente da estratégia de processamento de sinal usada.

Trypanosoma cruzi Promotes Transcriptomic Remodeling of the JAK/STAT Signaling and Cell Cycle Pathways in Myoblasts.

Chagas disease is responsible for more than 10,000 deaths per year and about 6 to 7 million infected people worldwide. In its chronic stage, patients can develop mega-colon, mega-esophagus, and cardiomyopathy. Differences in clinical outcomes may be determined, in part, by the genetic background of the parasite that causes Chagas disease. Trypanosoma cruzi has a high genetic diversity, and each group of strains may elicit specific pathological responses in the host. Conflicting results have been reported in studies using various combinations of mammalian host-T. cruzi strains. We previously profiled the transcriptomic signatures resulting from infection of L6E9 rat myoblasts with four reference strains of T. cruzi (Brazil, CL, Y, and Tulahuen). The four strains induced similar overall gene expression alterations in the myoblasts, although only 21 genes were equally affected by all strains. Cardiotrophin-like cytokine factor 1 (Clcf1) was one of the genes found to be consistently upregulated by the infection with all four strains of T. cruzi. This cytokine is a member of the interleukin-6 family that binds to glycoprotein 130 receptor and activates the JAK/STAT signaling pathway, which may lead to muscle cell hypertrophy. Another commonly upregulated gene was tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein theta (Ywhaq, 14-3-3 protein Θ), present in the Cell Cycle Pathway. In the present work, we reanalyzed our previous microarray dataset, aiming at understanding in more details the transcriptomic impact that each strain has on JAK/STAT signaling and Cell Cycle pathways. Using Pearson correlation analysis between the expression levels of gene pairs in biological replicas from each pathway, we determined the coordination between such pairs in each experimental condition and the predicted protein interactions between the significantly altered genes by each strain. We found that although these highlighted genes were similarly affected by all four strains, the downstream genes or their interaction partners were not necessarily equally affected, thus reinforcing the idea of the role of parasite background on host cell transcriptome. These new analyses provide further evidence to the mechanistic understanding of how distinct T. cruzi strains lead to diverse remodeling of host cell transcriptome.

Synthesis and trypanocidal activity of novel pyridinyl-1,3,4-thiadiazole derivatives.

Herein, we present the design, synthesis and trypanocidal evaluation of sixteen new 1,3,4-thiadiazole derivatives from N-aminobenzyl or N-arylhydrazone series. All derivatives were assayed against the trypomastigote form of Trypanosoma cruzi, showing IC50 values ranging from 3 to 226 µM, and a better trypanocidal profile was demonstrated for the 1,3,4-thiadiazole-N-arylhydrazones (3a-g). In this series, the 2-pyridinyl fragment bound to the imine subunit of the hydrazine moiety presented pharmacophoric behavior for trypanocidal activity. Compounds 2a, 11a and 3e presented remarkable activity and excellent selectivity indexes. Compound 2a was also active against the intracellular amastigote form of T. cruzi. Moreover, its corresponding hydrochloride, compound 11a, showed the most promising profile, producing phenotypic changes similar to those caused by posaconazole, a well-known inhibitor of sterol biosynthesis. Thus, 1,3,4-thiadiazole derivative 11a could be considered a good prototype for the development of new drug candidates for Chagas disease therapy.

Development of auditory and language skills in children using cochlear implants with two signal processing strategies.

INTRODUCTION: The increase in the spectral information offered by the sound processing strategy HiRes 120 has led to great expectations for the pediatric population. Due to a shorter duration of auditory deprivation and higher neural plasticity, children could benefit more substantially from the spectral information of this sound processing strategy. OBJECTIVE: To compare auditory and language skills in Brazilian children with cochlear implants using the HiRes and HiRes 120 sound processing strategies. METHODS: Thirty children, aged 1-3 years, with congenital hearing loss, were divided into two groups, according to the signal processing strategy adjusted at the time of the cochlear implant activation. The assessed children were matched according to chronological age and the time of the cochlear implant use. The auditory and language skills were evaluated longitudinally through the Infant-Toddler Meaningful Auditory Integration Scale and Production Infant Scale Evaluation, carried out before surgery, and 3, 6 and 12 months after device implantation. The Mann-Whitney test was applied for the comparison between the two groups with a 5% significance level. RESULTS: The findings indicated development of hearing and language skills in the first year of cochlear implant use; however, there was no statistically significant difference in the evolution of such skills due to the adjusted processing strategy in the activation of the cochlear implant electrodes. CONCLUSION: The development of auditory and language skills in the assessed children was similar during the entire study period, regardless of which signal processing strategy was used.

Trypanosoma cruzi down-regulates mechanosensitive proteins in cardiomyocytes.

BACKGROUND: Cardiac physiology depends on coupling and electrical and mechanical coordination through the intercalated disc. Focal adhesions offer mechanical support and signal transduction events during heart contraction-relaxation processes. Talin links integrins to the actin cytoskeleton and serves as a scaffold for the recruitment of other proteins, such as paxillin in focal adhesion formation and regulation. Chagasic cardiomyopathy is caused by infection by Trypanosoma cruzi and is a debilitating condition comprising extensive fibrosis, inflammation, cardiac hypertrophy and electrical alterations that culminate in heart failure. OBJECTIVES: Since mechanotransduction coordinates heart function, we evaluated the underlying mechanism implicated in the mechanical changes, focusing especially in mechanosensitive proteins and related signalling pathways during infection of cardiac cells by T. cruzi. METHODS: We investigated the effect of T. cruzi infection on the expression and distribution of talin/paxillin and associated proteins in mouse cardiomyocytes in vitro by western blotting, immunofluorescence and quantitative real-time polymerase chain reaction (qRT-PCR). FINDINGS: Talin and paxillin spatial distribution in T. cruzi-infected cardiomyocytes in vitro were altered associated with a downregulation of these proteins and mRNAs levels at 72 h post-infection (hpi). Additionally, we observed an increase in the activation of the focal adhesion kinase (FAK) concomitant with increase in ß-1-integrin at 24 hpi. Finally, we detected a decrease in the activation of FAK at 72 hpi in T. cruzi-infected cultures. MAIN CONCLUSION: The results suggest that these changes may contribute to the mechanotransduction disturbance evidenced in chagasic cardiomyopathy.

Trypanosoma cruzi down-regulates mechanosensitive proteins in cardiomyocytes

BACKGROUND Cardiac physiology depends on coupling and electrical and mechanical coordination through the intercalated disc. Focal adhesions offer mechanical support and signal transduction events during heart contraction-relaxation processes. Talin links integrins to the actin cytoskeleton and serves as a scaffold for the recruitment of other proteins, such as paxillin in focal adhesion formation and regulation. Chagasic cardiomyopathy is caused by infection by Trypanosoma cruzi and is a debilitating condition comprising extensive fibrosis, inflammation, cardiac hypertrophy and electrical alterations that culminate in heart failure. OBJECTIVES Since mechanotransduction coordinates heart function, we evaluated the underlying mechanism implicated in the mechanical changes, focusing especially in mechanosensitive proteins and related signalling pathways during infection of cardiac cells by T. cruzi. METHODS We investigated the effect of T. cruzi infection on the expression and distribution of talin/paxillin and associated proteins in mouse cardiomyocytes in vitro by western blotting, immunofluorescence and quantitative real-time polymerase chain reaction (qRT-PCR). FINDINGS Talin and paxillin spatial distribution in T. cruzi-infected cardiomyocytes in vitro were altered associated with a downregulation of these proteins and mRNAs levels at 72 h post-infection (hpi). Additionally, we observed an increase in the activation of the focal adhesion kinase (FAK) concomitant with increase in β-1-integrin at 24 hpi. Finally, we detected a decrease in the activation of FAK at 72 hpi in T. cruzi-infected cultures. MAIN CONCLUSION The results suggest that these changes may contribute to the mechanotransduction disturbance evidenced in chagasic cardiomyopathy.

Improved sealing ability promoted by calcium silicate-based root canal sealers

Aim: This study investigated the sealing ability produced by AH Plus (Dentsply DeTrey, Konstaz, Germany), EndoSequence BC sealer (Brasseler USA, Savannah, GA, USA), GuttaFlow (Coltène/Whaledent, Altstätten, France) and MTA Fillapex (Angelus, Londrina, Brazil). Methdos: A total of forty-six single-root human canines were prepared and randomly divided into four experimental groups (n=10): MTA Fillapex, EndoSequence BC Sealer, AH Plus or GuttaFlow. Teeth with intact crowns served as negative controls (n=3) and teeth filled with only gutta-percha served as positive controls (n=3). Teeth were mounted in a two-chamber apparatus and exposed to Enterococcus faecalis. The number of days over a 60-days period was recorded for the appearance of turbidity in the lower chamber. Kaplan-Meier method was used to estimate the survival curves. The nonparametric log-rank test was used to compare the survival curves using a significance level of 0.05. Results: The results at the end of the observation time were further analyzed by chi-square testing. All positive controls leaked within 24 h, whereas none of the negative controls leaked after 60 days. EndoSequence BC Sealer and MTA Fillapex had significant less bacterial leakage (P<0.05) than the other tested sealers. No significant difference between AH Plus and GuttaFlow was observed (P>0.05). Conclusion: In conclusion, calcium-silicate based root canal sealers promoted improved sealing ability when compared to other endodontic sealers

Relatos de médicos sobre a experiência do processo de morrer e a morte de seus pacientes / Medical reports about the experience of the dying process and the death of his/her patients

A morte é concebida diferentemente em cada cultura, e, sendo assim, cada povo tem uma forma de abordar, estudar e debater seus conceitos. Entre os médicos não é diferente, pois ela faz parte de sua profissão, principalmente nas especialidades que lidam com as situações terminais. O objetivo dessa pesquisa foi compreender como os médicos, de diferentes especialidades, encaram a morte iminente ou constatada de pacientes e identificar se houve, na sua formação acadêmica, algum preparo para isso. Esta pesquisa foi descritiva, de abordagem qualitativa e de corte transversal. Através de uma entrevista semiestruturada, cada um dos 9 profissionais abordados expressou o seu entendimento sobre a morte e relatou seu preparo para lidar com ela. Para análise dos dados quantificou-se gênero, idade e tempo de formação. Para as respostas abertas, criaram-se três categorias: 1) como cada profissional encara a morte do seu paciente; 2) se houve algum preparo para lidar com a morte de seus pacientes (formal ou não); 3) renovação de energias.

Death is conceived differently in every culture, every people has a way to address, study and discuss their concepts. Among physicians, it is no different, for it is part of their profession, specifically the specialties that deal with the terminal patients. The objective of this research was to understand how physicians from different specialties, face imminent or established patient's death and identify whether there was, in his academic background, some preparation for it. This research is descriptive qualitative approach, cross-sectional. Through a semi-structured interview, 9 professionals from different specialties expressed, in different ways, how they understand death and how they prepare to deal with it. For the later analysis, it was quantified: gender, age and training time. And as for the opened answers, 3 categories were created: 1) How each professional deal with the death of their patient; 2) if there was any kind preparation to deal with their patient's death (formal or not); 3) Energy's renovation.

TGF-ß receptor type II costameric localization in cardiomyocytes and host cell TGF-ß response is disrupted by Trypanosoma cruzi infection.

Transforming growth factor beta (TGF-ß) cytokine is involved in Chagas disease establishment and progression. Since Trypanosoma cruzi can modulate host cell receptors, we analysed the TGF-ß receptor type II (TßRII) expression and distribution during T. cruzi - cardiomyocyte interaction. TßRII immunofluorescent staining revealed a striated organization in cardiomyocytes, which was co-localized with vinculin costameres and enhanced (38%) after TGF-ß treatment. Cytochalasin D induced a decrease of 45·3% in the ratio of cardiomyocytes presenting TßRII striations, demonstrating an association of TßRII with the cytoskeleton. Western blot analysis showed that cytochalasin D significantly inhibited Smad 2 phosphorylation and fibronectin stimulation after TGF-ß treatment in cardiomyocytes. Trypanosoma cruzi infection elicited a decrease of 79·8% in the frequency of cardiomyocytes presenting TßRII striations, but did not interfere significantly in its expression. In addition, T. cruzi-infected cardiomyocytes present a lower response to exogenous TGF-ß, showing no enhancement of TßRII striations and a reduction of phosphorylated Smad 2, with no significant difference in TßRII expression when compared to uninfected cells. Together, these results suggest that the co-localization of TßRII with costameres is important in activating the TGF-ß signalling cascade, and that T. cruzi-derived cytoskeleton disorganization could result in altered or low TGF-ß response in infected cardiomyocytes.

Parents' perception of health-related quality of life in children and adolescents with excess weight / Percepção dos pais acerca da qualidade de vida relacionada à saúde de crianças e adolescentes com excesso de peso

ABSTRACT OBJECTIVE: To evaluate the perception of parents or caregivers on the health-related quality of life (HRQOL) of children/adolescents with overweight/obesity and possible factors associated with this perception. METHODS: This was a cross-sectional study involving 297 caregivers of children and adolescents with normal weight (n = 170) and with overweight/obesity (n = 127), from public and private schools in the study municipality. HRQOL scores obtained through the Child Health Questionnaire - Parent Form 50 (CHQ-PF50) were compared according to the nutritional status and gender of the children/adolescents. Multiple regression analysis was used to determine the predictive value of studied variables for the variation in HRQOL scores. RESULTS: Parents of children/adolescents with overweight/obesity attributed lower HRQOL scores to their children in the following domains: physical functioning (p < 0.01; d = 0.49), self-esteem (p < 0.01; d = 0.38), parental impact-emotional (p < 0.05; d= 0.29), family cohesion (p < 0.05; d = 0.26), physical summary score (p < 0.05;d = 0.29), and psychosocial summary score (p < 0.05; d = 0.25). In the multiple regression models, the variables with the highest contribution to the variation in HRQOL scores were: in the physical functioning domain, parental impact-time (ß = 0.23; p < 0.05); self-esteem, nutritional status (ß = -0.18;p ≤ 0.01); emotional impact on parents, impact on parents' time (ß = 0.31; p < 0.05); and in family cohesion, global behavior (ß = 0.30;p < 0.05). CONCLUSIONS: A negative impact on HRQOL of children/adolescents with overweight/obesity was observed in the physical and psychosocial aspects. The nutritional status was the variable with the greatest contribution for the assessment the self-esteem of children and adolescents in this study.

RESUMO OBJETIVO: Avaliar a percepção dos pais ou cuidadores a respeito da qualidade de vida relacionada à saúde (QVRS) de crianças/adolescentes com sobrepeso/obesidade e os possíveis fatores associados a essa percepção. MÉTODOS: Estudo transversal com a participação de 297 cuidadores de crianças e adolescentes eutróficos (n = 170) e com sobrepeso/obesidade (n = 127), provenientes de escolas públicas e privadas do município do estudo. Escores de QVRS obtidos por meio do Child Health Questionnaire - Parent Form 50 (CHQ-PF50) foram comparados de acordo com o estado nutricional e sexo das crianças/adolescentes. Análises de regressão múltipla foram usadas para determinar o valor preditivo das variáveis estudadas para a variação dos escores de QVRS. RESULTADOS: Pais de crianças/adolescentes com sobrepeso/obesidade atribuíram menores escores de QVRS para seus filhos nos domínios: função física (p < 0,01; d = 0,49), autoestima (p < 0,01; d = 0,38), impacto emocional dos pais (p < 0,05; d = 0,29), coesão familiar (p < 0,05; d = 0,26), sumário do escore físico (p < 0,05; d = 0,29) e sumário do escore psicossocial (p < 0,05; d = 0,25). Nos modelos de regressão múltipla, as variáveis com maior contribuição para a variação dos escores de QVRS foram: no domínio função física, impacto no tempo dos pais (ß = 0,23; p < 0,05); autoestima, estado nutricional (ß = -0,18; p ≤ 0,01); impacto emocional nos pais, impacto no tempo dos pais (ß = 0,31; p < 0,05); coesão familiar, comportamento global (ß = 0,30; p < 0,05). CONCLUSÕES: Há impacto negativo na QVRS de crianças/adolescentes com sobrepeso/obesidade em aspectos físicos e psicossociais. O estado nutricional foi a variável de maior contribuição para a avaliação da autoestima das crianças e adolescentes do presente estudo.

Parents' perception of health-related quality of life in children and adolescents with excess weight.

OBJECTIVE: To evaluate the perception of parents or caregivers on the health-related quality of life (HRQOL) of children/adolescents with overweight/obesity and possible factors associated with this perception. METHODS: This was a cross-sectional study involving 297 caregivers of children and adolescents with normal weight (n=170) and with overweight/obesity (n=127), from public and private schools in the study municipality. HRQOL scores obtained through the Child Health Questionnaire - Parent Form 50 (CHQ-PF50) were compared according to the nutritional status and gender of the children/adolescents. Multiple regression analysis was used to determine the predictive value of studied variables for the variation in HRQOL scores. RESULTS: Parents of children/adolescents with overweight/obesity attributed lower HRQOL scores to their children in the following domains: physical functioning (p<0.01; d=0.49), self-esteem (p<0.01; d=0.38), parental impact-emotional (p<0.05; d=0.29), family cohesion (p<0.05; d=0.26), physical summary score (p<0.05; d=0.29), and psychosocial summary score (p<0.05; d=0.25). In the multiple regression models, the variables with the highest contribution to the variation in HRQOL scores were: in the physical functioning domain, parental impact-time (ß=0.23; p<0.05); self-esteem, nutritional status (ß=-0.18; p≤0.01); emotional impact on parents, impact on parents' time (ß=0.31; p<0.05); and in family cohesion, global behavior (ß=0.30; p<0.05). CONCLUSIONS: A negative impact on HRQOL of children/adolescents with overweight/obesity was observed in the physical and psychosocial aspects. The nutritional status was the variable with the greatest contribution for the assessment the self-esteem of children and adolescents in this study.

HiFocus Helix™ electrode insertion: surgical approach.

BACKGROUND: Cochlear implants have been used for almost 30 years as a device for the rehabilitation of individuals with severe-to-profound hearing loss. One of the important aspects of cochlear implantation is the type of electrode selected and proper insertion of the electrode array in scala tympani to minimize cochlear damage. The HiFocus Helix™ electrode is a precurved design aimed at placing the electrode contacts close to the spiral ganglion cells in the modiolus. The prescribed insertion techniques are intended to minimize the likelihood of damage to the basilar membrane or lateral wall of the cochlea. CASE PRESENTATION: To describe the first insertion of a HiFocus Helix™ electrode in Brazil exposing surgical particularities and device details in a patient with profound hearing loss, due to Mondini's dysplasia. CONCLUSION: No problems were encountered during the surgical procedure. The patient experienced improvement in hearing thresholds and speech perception. The HiFocus Helix™ electrode proved easy to insert and provided expected hearing benefits for the patient. This manuscript indicates that the HiResolution™ Bionic Ear System with HiFocus Helix™ electrode comprise a cochlear implant system that is practical and beneficial for the treatment of severe-to-profound hearing loss.

Falhas dos componentes externos do implante coclear: influência da idade e do tempo de uso / Repair issues associated with cochlear implants external components: the influence of age and time of use

OBJETIVO: identificar quais são as falhas técnicas mais frequentes dos componentes externos do implante coclear e verificar a influência da idade do usuário e o tempo de uso do dispositivo nestas falhas. MÉTODOS: estudo retrospectivo, por meio da análise de 128 prontuários da Associação dos Deficientes Auditivos, Pais, Amigos e Usuários de Implante Coclear, que apresentaram alguma queixa no funcionamento do componente externo, no período de junho a agosto de 2011. Para análise dos dados foi realizada a análise descritiva dos componentes que apresentaram falha técnica, bem como das variáveis estudadas. Posteriormente, foi aplicada a estatística inferencial por meio do teste Kruskal-Wallis para verificar a influência da idade do paciente e tempo de uso do implante coclear com a falha apresentada, com nível de significância p< 0,05. RESULTADOS: os componentes que apresentaram mais falhas foram: antena externa (29,7%), seguida do compartimento de bateria (23,4%), processador de fala (14,10%), microfone (10,90%), cabos de transmissão (9,40%), controlador (6,30%), baterias (5,50%) e carregador de baterias (0,80%). Não foi verificada influência do tempo de uso do implante coclear e a idade cronológica do paciente em relação às falhas apresentadas no presente estudo. CONCLUSÃO: os achados obtidos a partir do presente estudo podem auxiliar os profissionais da área na orientação sobre a manutenção destes componentes, mas as tendências aqui apresentadas não podem ser generalizadas para todo território nacional, pois estas foram analisadas em um número limitado de casos. .

PURPOSE: to identify what are the most frequent technical flaws of the external components of the cochlear implant and verify the influence of age and the time of use of the device in these failures. METHODS: retrospective study. It was analyzed 128 medical records of the Association of Deaf Parents, Friends and Cochlear Implant Users, which presented a complaint in the functioning of the external component for the period from June to August 2011. For analysis of data was carried out the descriptive analysis of failed components, as well as the variables studied. It was applied to an inferential statistics through the Kruskal-Wallis test to analyze the influence of patient’s age and time of use of the cochlear implant with failure, with significance level p <0.05. RESULTS: the components that were more failure was the external antenna (29,7%), followed by the battery compartment (23,4%), speech processor (14,10%), microphone (10,90%), transmission cables (9,40%), controller (6,30%), batteries (5,50) and battery charge (0,80%). Has not been verified influence of time of use of the cochlear implant and the chronological age of the patient in relation to failures presented in this study. CONCLUSION: the findings obtained from this study can assist professionals in guidance on the maintenance of these components, but the trends presented here cannot be generalized to the entire national territory, as these were analyzed in a limited number of cases. .