Specific Recognition of ß-Galactofuranose-Containing Glycans of Synthetic Neoglycoproteins by Sera of Chronic Chagas Disease Patients.

Chagas disease (CD) can be accurately diagnosed by detecting Trypanosoma cruzi in patients' blood using polymerase chain reaction (PCR). However, parasite-derived biomarkers are of great interest for the serological diagnosis and early evaluation of chemotherapeutic efficacy when PCR may fail, owing to a blood parasite load below the method's limit of detection. Previously, we focused on the detection of specific anti-α-galactopyranosyl (α-Gal) antibodies in chronic CD (CCD) patients elicited by α-Gal glycotopes copiously expressed on insect-derived and mammal-dwelling infective parasite stages. Nevertheless, these stages also abundantly express cell surface glycosylphosphatidylinositol (GPI)-anchored glycoproteins and glycoinositolphospholipids (GIPLs) bearing nonreducing terminal ß-galactofuranosyl (ß-Galf) residues, which are equally foreign to humans and, therefore, highly immunogenic. Here we report that CCD patients' sera react specifically with synthetic ß-Galf-containing glycans. We took a reversed immunoglycomics approach that entailed: (a) Synthesis of T. cruzi GIPL-derived Galfß1,3Manpα-(CH2)3SH (glycan G29SH) and Galfß1,3Manpα1,2-[Galfß1,3]Manpα-(CH2)3SH (glycan G32SH); and (b) preparation of neoglycoproteins NGP29b and NGP32b, and their evaluation in a chemiluminescent immunoassay. Receiver-operating characteristic analysis revealed that NGP32b can distinguish CCD sera from sera of healthy individuals with 85.3% sensitivity and 100% specificity. This suggests that Galfß1,3Manpα1,2-[Galfß1,3]Manpα is an immunodominant glycotope and that NGP32b could potentially be used as a novel CCD biomarker.

TcI Isolates of Trypanosoma cruzi Exploit the Antioxidant Network for Enhanced Intracellular Survival in Macrophages and Virulence in Mice.

Trypanosoma cruzi species is categorized into six discrete typing units (TcI to TcVI) of which TcI is most abundantly noted in the sylvatic transmission cycle and considered the major cause of human disease. In our study, the TcI strains Colombiana (COL), SylvioX10/4 (SYL), and a cultured clone (TCC) exhibited different biological behavior in a murine model, ranging from high parasitemia and symptomatic cardiomyopathy (SYL), mild parasitemia and high tissue tropism (COL), to no pathogenicity (TCC). Proteomic profiling of the insect (epimastigote) and infective (trypomastigote) forms by two-dimensional gel electrophoresis/matrix-assisted laser desorption ionization-time of flight mass spectrometry, followed by functional annotation of the differential proteome data sets (≥2-fold change, P < 0.05), showed that several proteins involved in (i) cytoskeletal assembly and remodeling, essential for flagellar wave frequency and amplitude and forward motility of the parasite, and (ii) the parasite-specific antioxidant network were enhanced in COL and SYL (versus TCC) trypomastigotes. Western blotting confirmed the enhanced protein levels of cytosolic and mitochondrial tryparedoxin peroxidases and their substrate (tryparedoxin) and iron superoxide dismutase in COL and SYL (versus TCC) trypomastigotes. Further, COL and SYL (but not TCC) were resistant to exogenous treatment with stable oxidants (H2O2 and peroxynitrite [ONOO(-)]) and dampened the intracellular superoxide and nitric oxide response in macrophages, and thus these isolates escaped from macrophages. Our findings suggest that protein expression conducive to increase in motility and control of macrophage-derived free radicals provides survival and persistence benefits to TcI isolates of T. cruzi.

Workshop on pediatric trauma care: low-cost simulation.

OBJECTIVE: to assess nursing students' and nurses' knowledge, satisfaction and self-confidence after a theoretical workshop on emergency care for traumatized children and clinical simulation. METHODS: a quasi-experimental study, carried out with nursing students and nurses residing at a public university in southern Brazil. A workshop on pediatric trauma care was created and a mannequin was created for simulations. A knowledge pre-test and post-test and the Student Satisfaction and Self-Confidence in Learning instrument were applied to measure satisfaction and self-confidence in learning. For analysis, descriptive statistics and the Wilcoxon test were used to compare means before and after intervention. RESULTS: the difference between misses and hits was statistically significant (p<0.005), demonstrating an increase in participants' knowledge after the workshop. Satisfaction and self-confidence were demonstrated in the instrument's high scores. CONCLUSIONS: the effectiveness of the workshop in teaching-learning emergency care for pediatric trauma was demonstrated.

Prognostic Performance of Peripheral Blood Biomarkers in Identifying Seropositive Individuals at Risk of Developing Clinically Symptomatic Chagas Cardiomyopathy.

Biomarkers for prognosis-based detection of Trypanosoma cruzi-infected patients presenting no clinical symptoms to cardiac Chagas disease (CD) are not available. In this study, we examined the performance of seven biomarkers in prognosis and risk of symptomatic CD development. T. cruzi-infected patients clinically asymptomatic (C/A; n = 30) or clinically symptomatic (C/S; n = 30) for cardiac disease and humans who were noninfected and healthy (N/H; n = 24) were enrolled (1 - ß = 80%, α = 0.05). Serum, plasma, and peripheral blood mononuclear cells (PBMCs) were analyzed for heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1), vimentin, poly(ADP-ribose) polymerase (PARP1), 8-hydroxy-2-deoxyguanosine (8-OHdG), copeptin, endostatin, and myostatin biomarkers by enzyme-linked immunosorbent assay (ELISA) and Western blotting. Secreted hnRNPA1, vimentin, PARP1, 8-OHdG, copeptin, and endostatin were increased by 1.4- to 7.0-fold in CD subjects versus N/H subjects (P < 0.001) and showed excellent predictive value in identifying the occurrence of infection (area under the receiver operating characteristic [ROC] curve [AUC], 0.935 to 0.999). Of these, vimentin, 8-OHdG, and copeptin exhibited the best performance in prognosis of C/S (versus C/A) CD, determined by binary logistic regression analysis with the Cox and Snell test (R2C&S = 0.492 to 0.688). A decline in myostatin and increase in hnRNPA1 also exhibited good predictive value in identifying C/S and C/A CD status, respectively. Furthermore, circulatory 8-OHdG (Wald χ2 = 15.065), vimentin (Wald χ2 = 14.587), and endostatin (Wald χ2 = 17.902) levels exhibited a strong association with changes in left ventricular ejection fraction and diastolic diameter (P = 0.001) and predicted the risk of cardiomyopathy development in CD patients. We have identified four biomarkers (vimentin, 8-OHdG, copeptin, and endostatin) that offer excellent value in prognosis and risk of symptomatic CD development. Decline in these four biomarkers and increase in hnRNPA1 would be useful in monitoring the efficacy of therapies and vaccines in halting CD. IMPORTANCE There is a lack of validated biomarkers for diagnosis of T. cruzi-infected individuals at risk of developing heart disease. Of the seven potential biomarkers that were screened, vimentin, 8-OHdG, copeptin, and endostatin exhibited excellent performance in distinguishing the clinical severity of Chagas disease. A decline in these four biomarkers can also be used for monitoring the therapeutic responses of infected patients to established or newly developed drugs and vaccines and precisely inform the patients about their progress. These biomarkers can easily be screened using the readily available plasma/serum samples in the clinical setting by an ELISA that is inexpensive, fast, and requires low-tech resources at the facility, equipment, and personnel levels.

Workshop on pediatric trauma care: low-cost simulation / Workshop sobre atención de trauma pediátrico: simulación de bajo costo / Workshop sobre atendimento de trauma pediátrico: simulação de baixo custo

ABSTRACT Objective: to assess nursing students' and nurses' knowledge, satisfaction and self-confidence after a theoretical workshop on emergency care for traumatized children and clinical simulation. Methods: a quasi-experimental study, carried out with nursing students and nurses residing at a public university in southern Brazil. A workshop on pediatric trauma care was created and a mannequin was created for simulations. A knowledge pre-test and post-test and the Student Satisfaction and Self-Confidence in Learning instrument were applied to measure satisfaction and self-confidence in learning. For analysis, descriptive statistics and the Wilcoxon test were used to compare means before and after intervention. Results: the difference between misses and hits was statistically significant (p<0.005), demonstrating an increase in participants' knowledge after the workshop. Satisfaction and self-confidence were demonstrated in the instrument's high scores. Conclusions: the effectiveness of the workshop in teaching-learning emergency care for pediatric trauma was demonstrated.

RESUMEN Objetivo: evaluar el conocimiento, la satisfacción y la confianza en sí mismos de estudiantes y enfermeros de enfermería luego de un workshop teórico sobre atención de emergencia al niño traumatizado y simulación clínica. Métodos: estudio cuasiexperimental, realizado con estudiantes de enfermería y enfermeros residentes en una universidad pública del sur de Brasil. Se creó un workshop de atención al trauma pediátrico y se creó un maniquí para las simulaciones. Para medir la satisfacción y la autoconfianza en el aprendizaje se aplicó un preprueba y posprueba de conocimientos y el instrumento de Satisfacción Estudiantil y Autoconfianza en el Aprendizaje. Para el análisis se utilizó estadística descriptiva y la prueba de Wilcoxon para comparar medias antes y después de la intervención. Resultados: la diferencia entre errores y aciertos fue estadísticamente significativa (p<0,005), demostrando un aumento en el conocimiento de los participantes después del workshop. La satisfacción y la confianza en uno mismo se demostraron en las altas puntuaciones del instrumento. Conclusiones: se demostró la efectividad del workshop en la enseñanza-aprendizaje de la atención de emergencia en trauma pediátrico.

RESUMO Objetivo: avaliar conhecimento, satisfação e autoconfiança de estudantes de enfermagem e enfermeiros após workshop teórico sobre atendimento emergencial à criança traumatizada e simulação clínica. Métodos: estudo quase-experimental, realizado com estudantes de enfermagem e enfermeiros residentes em uma universidade pública no sul do Brasil. Foi construído um workshop sobre atendimento de trauma pediátrico e confeccionado um manequim para as simulações. Aplicaram-se pré-teste e pós-teste de conhecimento e o instrumento Student Satisfaction and Self-Confidence in Learning para mensurar a satisfação e autoconfiança na aprendizagem. Para análise, utilizaram-se a estatística descritiva e o teste de Wilcoxon para comparação das médias antes e após a intervenção. Resultados: a diferença entre erros e acertos foi estatisticamente significativa (p<0,005), demonstrando aumento do conhecimento dos participantes após o workshop. A satisfação e a autoconfiança foram demonstradas nos altos escores do instrumento. Conclusões: evidenciou-se a efetividade do workshop no ensino-aprendizagem do atendimento emergencial ao trauma pediátrico.

Potential Utility of Protein Targets of Cysteine-S-Nitrosylation in Identifying Clinical Disease Status in Human Chagas Disease.

Trypanosoma cruzi (Tc) infection causes Chagas disease (ChD) presented by dilated cardiomyopathy and heart failure. During infection, oxidative and nitrosative stresses are elicited by the immune cells for control the pathogen; however, excess nitric oxide and superoxide production can result in cysteine S-nitrosylation (SNO) of host proteins that affects cellular homeostasis and may contribute to disease development. To identify the proteins with changes in SNO modification levels as a hallmark of ChD, we obtained peripheral blood mononuclear cells (PBMC) from seronegative, normal healthy (NH, n = 30) subjects, and from seropositive clinically asymptomatic (ChD CA, n = 25) or clinically symptomatic (ChD CS, n = 28) ChD patients. All samples were treated (Asc+) or not-treated (Asc-) with ascorbate (reduces nitrosylated thiols), labeled with the thiol-labeling BODIPY FL-maleimide dye, resolved by two-dimensional electrophoresis (total 166 gels), and the protein spots that yielded significant differences in abundance or SNO level at p-value of ≤ 0.05 t-test/Welch/BH were identified by MALDI-TOF/TOF MS or OrbiTrap LC-MS/MS. Targeted analysis of a new cohort of PBMC samples (n = 10-14/group) was conducted to verify the differential abundance/SNO levels of two of the proteins in ChD (vs. NH) subjects. The multivariate adaptive regression splines (MARS) modeling, comparing differences in relative SNO level (Asc-/Asc+ ratio) of the protein spots between any two groups yielded SNO biomarkers that exhibited ≥90% prediction success in classifying ChD CA (582-KRT1 and 884-TPM3) and ChD CS (426-PNP, 582-KRT1, 486-ALB, 662-ACTB) patients from NH controls. Ingenuity Pathway Analysis (IPA) of the SNO proteome dataset normalized to changes in protein abundance suggested the proteins belonging to the signaling networks of cell death and the recruitment and migration of immune cells were most affected in ChD CA and ChD CS (vs. NH) subjects. We propose that SNO modification of the select panel of proteins identified in this study have the potential to identify ChD severity in seropositive individuals exposed to Tc infection.

Immune complexes in chronic Chagas disease patients are formed by exovesicles from Trypanosoma cruzi carrying the conserved MASP N-terminal region.

The exovesicles (EVs) are involved in pathologic host-parasite immune associations and have been recently used as biomarkers for diagnosis of infectious diseases. The release of EVs by Trypanosoma cruzi, the causative agent of Chagas disease, has recently been described, with different protein cargoes including the MASP multigene family of proteins MASPs are specific to this parasite and characterized by a conserved C-terminal (C-term) region and an N-terminal codifying for a signal peptide (SP). In this investigation, we identified immature MASP proteins containing the MASP SP in EVs secreted by the infective forms of the parasite. Those EVs are responsible for the formation of immune complexes (ICs) containing anti-MASP SP IgGs in patients with different (cardiac, digestive and asymptomatic) chronic Chagas disease manifestations. Moreover, purified EVs as well as the MASP SP inhibit the action of the complement system and also show a significant association with the humoral response in patients with digestive pathologies. These findings reveal a new route for the secretion of MASP proteins in T. cruzi, which uses EVs as vehicles for immature and misfolded proteins, forming circulating immune complexes. Such complexes could be used in the prognosis of digestive pathologies of clinical forms of Chagas disease.

Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients.

The discovery of T cell epitopes is essential not only for gaining knowledge about host response to infectious disease but also for the development of immune-intervention strategies. In Chagas disease, given the size and complexity of the Trypanosoma cruzi proteome and its interaction with the host's immune system, the fine specificity of T cells has not been extensively studied yet, and this is particularly true for the CD4+ T cell compartment. The aim of the present work was to optimize a protocol for the generation of parasite-specific memory T cell lines, representative of their in vivo precursor populations and capable of responding to parasite antigens after long-term culture. Accordingly, peripheral blood mononuclear cells (PBMC) from both chronic asymptomatic and cardiac patients, and from non-infected individuals, underwent different in vitro culture and stimulation conditions. Subsequently, cells were tested for their capacity to respond against T. cruzi lysate by measuring [3H]-thymidine incorporation and interferon-γ and GM-CSF secretion. Results allowed us to adjust initial T. cruzi lysate incubation time as well as the number of expansions with phytohemagglutinin (PHA) and irradiated allogeneic PBMC prior to specificity evaluation. Moreover, our data demonstrated that parasite specific T cells displayed a clear and strong activation by using T. cruzi lysate pulsed, Epstein-Barr virus (EBV)-transformed human B lymphocytes (B-LCL), as autologous antigen presenting cells. Under these culture conditions, we generated a clone from an asymptomatic patient's memory CD4+ T cells which responded against epimastigote and trypomastigote protein lysate. Our results describe a culture method for isolating T. cruzi specific T cell clones from patients with Chagas disease, which enable the acquisition of information on functionality and specificity of individual T cells.

Gene Expression Profiling and Functional Characterization of Macrophages in Response to Circulatory Microparticles Produced during Trypanosoma cruzi Infection and Chagas Disease.

BACKGROUND: Chronic inflammation and oxidative stress are hallmarks of chagasic cardiomyopathy (CCM). In this study, we determined if microparticles (MPs) generated during Trypanosoma cruzi (Tc) infection carry the host's signature of the inflammatory/oxidative state and provide information regarding the progression of clinical disease. METHODS: MPs were harvested from supernatants of human peripheral blood mononuclear cells in vitro incubated with Tc (control: LPS treated), plasma of seropositive humans with a clinically asymptomatic (CA) or symptomatic (CS) disease state (vs. normal/healthy [NH] controls), and plasma of mice immunized with a protective vaccine before challenge infection (control: unvaccinated/infected). Macrophages (mφs) were incubated with MPs, and we probed the gene expression profile using the inflammatory signaling cascade and cytokine/chemokine arrays, phenotypic markers of mφ activation by flow cytometry, cytokine profile by means of an ELISA and Bioplex assay, and oxidative/nitrosative stress and mitotoxicity by means of colorimetric and fluorometric assays. RESULTS: Tc- and LPS-induced MPs stimulated proliferation, inflammatory gene expression profile, and nitric oxide (∙NO) release in human THP-1 mφs. LPS-MPs were more immunostimulatory than Tc-MPs. Endothelial cells, T lymphocytes, and mφs were the major source of MPs shed in the plasma of chagasic humans and experimentally infected mice. The CS and CA (vs. NH) MPs elicited >2-fold increase in NO and mitochondrial oxidative stress in THP-1 mφs; however, CS (vs. CA) MPs elicited a more pronounced and disease-state-specific inflammatory gene expression profile (IKBKB, NR3C1, and TIRAP vs. CCR4, EGR2, and CCL3), cytokine release (IL-2 + IFN-γ > GCSF), and surface markers of mφ activation (CD14 and CD16). The circulatory MPs of nonvaccinated/infected mice induced 7.5-fold and 40% increases in ∙NO and IFN-γ production, respectively, while these responses were abolished when RAW264.7 mφs were incubated with circulatory MPs of vaccinated/infected mice. CONCLUSION: Circulating MPs reflect in vivo levels of an oxidative, nitrosative, and inflammatory state, and have potential utility in evaluating disease severity and the efficacy of vaccines and drug therapies against CCM.

In Vitro Human Umbilical Vein Endothelial Cells Response to Ionic Dissolution Products from Lithium-Containing 45S5 Bioactive Glass.

Since lithium (Li⁺) plays roles in angiogenesis, the localized and controlled release of Li⁺ ions from bioactive glasses (BGs) represents a promising alternative therapy for the regeneration and repair of tissues with a high degree of vascularization. Here, microparticles from a base 45S5 BG composition containing (wt %) 45% SiO2, 24.5% Na2O, 24.5% CaO, and 6% P2O5, in which Na2O was partially substituted by 5% Li2O (45S5.5Li), were obtained. The results demonstrate that human umbilical vein endothelial cells (HUVECs) have greater migratory and proliferative response and ability to form tubules in vitro after stimulation with the ionic dissolution products (IDPs) of the 45S5.5Li BG. The results also show the activation of the canonical Wnt/ß-catenin pathway and the increase in expression of proangiogenic cytokines insulin like growth factor 1 (IGF1) and transforming growth factor beta (TGFß). We conclude that the IDPs of 45S5.5Li BG would act as useful inorganic agents to improve tissue repair and regeneration, ultimately stimulating HUVECs behavior in the absence of exogenous growth factors.

Correction: Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients.

[This corrects the article DOI: 10.1371/journal.pone.0178380.].

Conhecimento teórico de graduandos sobre parada cardiorrespiratória no suporte básico de vida / Conocimientos teóricos de los estudiantes universitarios sobre la parada cardiorrespiratoria en soporte vital básico / Theoretical knowledge of undergraduates on cardiorespiratory arrest in basic life support

Objective identificar o conhecimento teórico de graduandos de cursos que não são da saúde sobre parada cardiorrespiratória no suporte básico de vida Method pesquisa descritiva e exploratória de natureza quantitativa realizada com graduandos de universidade pública localizada no Noroeste do Paraná, em 2019. Para coleta dos dados, utilizou-se questionário, contendo caracterização do sujeito e dez questões referentes ao reconhecimento e atendimento de situações de parada cardiorrespiratória e reanimação cardiopulmonar. Os dados foram tabulados e analisados. Results não se consideraram aptos para atendimento de evento de parada cardiorrespiratória 94,0% dos participantes; 92,6% não sabiam detectar essa condição; e 95,5% não souberam qual conduta adotar. Conclusion o conhecimento teórico de graduandos de cursos que não são da saúde sobre parada cardiorrespiratória foi insuficiente e é necessário treinamento para leigos em situações de emergência, para que o atendimento seja iniciado correta e imediatamente, sem postergar acionamento de serviço médico de emergência.

Objetivo identificar los conocimientos teóricos de los estudiantes universitarios de cursos no sanitarios sobre la parada cardiorrespiratoria en soporte vital básico. Método investigación descriptiva y exploratoria de carácter cuantitativo realizada con egresados de universidad pública ubicada en el Noroeste de Paraná, en 2019. Para la recolección de datos se utilizó un cuestionario, que contenía caracterización del sujeto y diez preguntas relacionadas con el reconocimiento y asistencia de situaciones de paro cardiorrespiratorio y reanimación cardiopulmonar. Los datos fueron tabulados y analizados. Resultados el 94,0% de los participantes no se consideraron aptos para el evento de paro cardiorrespiratorio; El 92,6% no sabía cómo detectar esta afección; y el 95,5% no sabía qué conducta adoptar. Conclusión los conocimientos teóricos de los estudiantes universitarios de cursos no sanitarios sobre parada cardiorrespiratoria fueron insuficientes y se requiere capacitación para los laicos en situaciones de emergencia, de modo que la atención se inicie de manera correcta e inmediata, sin posponer la activación del servicio médico de emergencia.

Objective to identify the theoretical knowledge of undergraduates of non-health courses about cardiorespiratory arrest on basic life support. Method descriptive and exploratory research of quantitative nature carried out with graduates of public university located in the Northwest of Paraná, in 2019. For data collection, a questionnaire was used, containing characterization of the subject and ten questions related to the recognition and attendance of situations of cardiorespiratory arrest and cardiopulmonary resuscitation. The data were tabulated and analyzed. Results 94.0% of the participants were not considered fit for cardiorespiratory arrest event; 92.6% did not know how to detect this condition; and 95.5% did not know which conduct to adopt. Conclusion the theoretical knowledge of undergraduates of non-health courses about cardiorespiratory arrest was insufficient and training is required for lay people in emergency situations, so that care is started correctly and immediately, without postponing emergency medical service activation.

S-Nitrosylation Proteome Profile of Peripheral Blood Mononuclear Cells in Human Heart Failure.

Nitric oxide (NO) protects the heart against ischemic injury; however, NO- and superoxide-dependent S-nitrosylation (S-NO) of cysteines can affect function of target proteins and play a role in disease outcome. We employed 2D-GE with thiol-labeling FL-maleimide dye and MALDI-TOF MS/MS to capture the quantitative changes in abundance and S-NO proteome of HF patients (versus healthy controls, n = 30/group). We identified 93 differentially abundant (59-increased/34-decreased) and 111 S-NO-modified (63-increased/48-decreased) protein spots, respectively, in HF subjects (versus controls, fold-change | ≥1.5|, p ≤ 0.05). Ingenuity pathway analysis of proteome datasets suggested that the pathways involved in phagocytes' migration, free radical production, and cell death were activated and fatty acid metabolism was decreased in HF subjects. Multivariate adaptive regression splines modeling of datasets identified a panel of proteins that will provide >90% prediction success in classifying HF subjects. Proteomic profiling identified ATP-synthase, thrombospondin-1 (THBS1), and vinculin (VCL) as top differentially abundant and S-NO-modified proteins, and these proteins were verified by Western blotting and ELISA in different set of HF subjects. We conclude that differential abundance and S-NO modification of proteins serve as a mechanism in regulating cell viability and free radical production, and THBS1 and VCL evaluation will potentially be useful in the prediction of heart failure.

Changes in Proteome Profile of Peripheral Blood Mononuclear Cells in Chronic Chagas Disease.

Trypanosoma cruzi (Tc) infection causes chagasic cardiomyopathy; however, why 30-40% of the patients develop clinical disease is not known. To discover the pathomechanisms in disease progression, we obtained the proteome signature of peripheral blood mononuclear cells (PBMCs) of normal healthy controls (N/H, n = 30) and subjects that were seropositive for Tc-specific antibodies, but were clinically asymptomatic (C/A, n = 25) or clinically symptomatic (C/S, n = 28) with cardiac involvement and left ventricular dysfunction. Protein samples were labeled with BODIPY FL-maleimide (dynamic range: > 4 orders of magnitude, detection limit: 5 f-mol) and resolved by two-dimensional gel electrophoresis (2D-GE). After normalizing the gel images, protein spots that exhibited differential abundance in any of the two groups were analyzed by mass spectrometry, and searched against UniProt human database for protein identification. We found 213 and 199 protein spots (fold change: |≥ 1.5|, p< 0.05) were differentially abundant in C/A and C/S individuals, respectively, with respect to N/H controls. Ingenuity Pathway Analysis (IPA) of PBMCs proteome dataset identified an increase in disorganization of cytoskeletal assembly and recruitment/activation and migration of immune cells in all chagasic subjects, though the invasion capacity of cells was decreased in C/S individuals. IPA predicted with high probability a decline in cell survival and free radical scavenging capacity in C/S (but not C/A) subjects. The MYC/SP1 transcription factors that regulate hypoxia and oxidative/inflammatory stress were predicted to be key targets in the context of control of Chagas disease severity. Further, MARS-modeling identified a panel of proteins that had >93% prediction success in classifying infected individuals with no disease and those with cardiac involvement and LV dysfunction. In conclusion, we have identified molecular pathways and a panel of proteins that could aid in detecting seropositive individuals at risk of developing cardiomyopathy.

Vitamin A deficiency induces prooxidant environment and inflammation in rat aorta.

We evaluated whether nutritional vitamin A deficiency generates oxidative stress and inflammation in aorta. Wistar male rats (21 days old) were given free access to a control (8 mg retinol as retinyl palmitate/kg) or a vitamin A- deficient diet for three months. One group of deficient animals was fed with the control diet fifteen days before sacrifice. Thiobarbituric acid-reactive substances (TBARS) and nitrite concentration where both analyzed in serum and aorta. Aorta Copper-Zinc Superoxide dismutase (CuZnSOD), Glutathion peroxidase (GPx) and Catalase (CAT) activities were measured. In addition, binding activity of the nuclear factor- kB (NF-kB), inducible and endothelial Nitric Oxide synthase (iNOS and eNOS, respectively) and Ciclooxygenase-2 (COX-2) expressions were determinated in aorta. Rats fed the vitamin A- deficient diet were characterized by sub-clinical plasma retinol concentration and showed increased serum and aorta concentrations of TBARS compared to controls. Lower than control activities of CuZnSOD, GPx, and CAT were observed in aorta of the vitamin A- deficient group. The binding activity of NF- kB was higher in vitamin A- deficient animals than controls. In addition, NO production evaluated as nitrite concentration increased in aorta and serum, associated with a higher expression of iNOS, eNOS and COX-2 in aorta of vitamin A-deficient rats. The incorporation of vitamin A into the diet of vitamin A-deficient rats reverted the changes observed in TBARS level, CuZnSOD and GPx activities, nitrite concentration and also, iNOS, eNOS and COX-2 expression. Prooxidant environment and inflammation are induced by vitamin A deficiency in rat aorta.

Association between ocular abnormalities and systemic diseases in Down Syndrome patients / Associação entre achados oftalmológicos e comorbidades em pacientes com Síndrome de Down

Abstract Objectives: Describe ocular findings and its correlation with systemic diseases in Down Syndrome (DS) pediatric patients. Methods: Quantitative and cross-sectional study of prevalence with children aged from 0 to 25 years. Standard ophthalmic examinations performed: visual acuity, slit lamp biomicroscopy, ocular motility, static refraction and indirect ophthalmoscopy. Ocular findings were associated with comorbidities available in pediatric records of patients in FURB Down Syndrome Outpatient clinic, in which they have regular follow-up. Results: A total of 76 patients were evaluated (33 males and 43 females). Of these, 72 patients (94.73%) had ocular abnormalities. Refractive errors were the most prevalent (94.73%), followed by alterations in indirect ophthalmoscopy (40.8%), biomicroscopy (15.8%), ocular motility (15.8%) and epiphora (9.2%). From the refractive changes 13.15% had myopia; 76.31% had hypermetropia and 47.36% had astigmatism. Sytemic abnormalities were observed in 73 children. The most prevalent was thyroid diseases presented in 65.79%, followed by heart disease 61.84%, gastrointestinal disease (15.79%); abdominal hernias (14.4%); respiratory changes (14.4%); genitourinary alterations (10.53%); musculoskeletal alterations (10.53%) and epilepsy (3.95%). There was statically significant association between the presence of myopia and hypothyroidism (p = 0.01); astigmatism and heart diseases (p = 0.003); and astigmatism and genitourinary alterations (p = 0.001). Conclusion: There was a high prevalence of ophthalmologic abnormalities in this study of children with Down Syndrome. Associations between myopia and hypothyroidism, astigmatism and heart diseases, and astigmatism and genitourinary disorders were found. More studies and increase of the sample are necessary to confirm the associations of ophthalmologic abnormalities with most common systemic diseases in this population.

Resumo Objetivos: Descrever as alterações oculares e sua correlação com outras comorbidades em pacientes pediátricos com Síndrome de Down (SD). Métodos: Estudo quantitativo de prevalência com delineamento transversal em crianças de 0 a 25 anos portadoras de SD. Realizados exames oftalmológicos de acuidade visual, biomicroscopia anterior, motilidade ocular, refração estática objetiva ou subjetiva conforme o nível de cooperação do paciente e oftalmoscopia indireta. Os achados foram correlacionados com as comorbidades disponíveis nos prontuários dos pacientes do Ambulatório de Síndrome de Down da FURB, no qual são acompanhados regularmente. Resultados: Foram avaliados 76 pacientes (33 do sexo masculino e 43 do sexo feminino). Dentre esses, 72 pacientes (94,73%) tiveram alterações oculares. Alterações refrativas foram as mais prevalentes (94,73%), seguidas de alterações na oftalmoscopia indireta (40,8%), biomicroscopia (15,8%), motilidade ocular (15,8%) e epífora (9,2%). Das alterações refrativas 13,15% tiveram miopia; 76,31% tiveram hipermetropia e 47,36% tiveram astigmatismo. Na amostra, 73 pacientes possuíam alguma comorbidade. A mais prevalente foi a alteração de tireoide, presente em 65,79% dos pacientes, seguido de alterações cardíacas (61,84%), alterações gastrointestinais (15,79%), hérnias abdominais (14,4%), alterações respiratórias (14,4%), alterações geniturinárias (10,53%), alterações osteomusculares (10,53%) e epilepsia (3,95%). Houve associações significativas entre miopia e hipotireoidismo (p = 0,01); astigmatismo e cardiopatias (p = 0,003); e astigmatismo e alterações geniturinárias (p = 0,001). Conclusão: Houve alta prevalência de alterações oftalmológicas na amostra. Foram encontradas associações entre miopia e hipotireoidismo, astigmatismo e cardiopatias, e astigmatismo e alterações geniturinárias. Mais estudos e aumento da amostra são necessários para confirmar os resultados das associações nessa população.

Angiogenic effects of ionic dissolution products released from a boron-doped 45S5 bioactive glass.

In regenerative medicine of vascularized tissues, there is a great interest in the use of biomaterials that are able to stimulate angiogenesis, a process necessary for rapid revascularization to allow the transport and exchange of oxygen, nutrients, growth factors and cells that take part in tissue repair and/or regeneration. An increasing number of publications have shown that bioactive glasses stimulate angiogenesis. Because it has been established that boron (B) may play a role in angiogenesis, the aim of this study was to assess the in vivo angiogenic effects of the ionic dissolution products that from a bioactive glass (BG) in the 45S5 system doped with 2 wt% B2O3 (45S5.2B). The pro-angiogenic capacity of 45S5.2B BG was assessed on the vasculature of the embryonic quail chorioallantoic membrane (CAM). Ionic dissolution products from 45S5.2B BG increased angiogenesis. This is quantitatively evidenced by the greater expression of integrin αvß3 and higher vascular density in the embryonic quail CAM. The response observed at 2 and 5 days post-treatment was equivalent to that achieved by applying 10 µg mL-1 of basic fibroblast growth factor. These results show that the ionic dissolution products released from the bioactive glass 45S5.2B stimulate angiogenesis in vivo. The effects observed are attributed to the presence the ionic dissolution products, which contained 160 ± 10 µM borate.

Innate immune responses and antioxidant/oxidant imbalance are major determinants of human Chagas disease.

BACKGROUND: We investigated the pathological and diagnostic role of selected markers of inflammation, oxidant/antioxidant status, and cellular injury in human Chagas disease. METHODS: Seropositive/chagasic subjects characterized as clinically-symptomatic or clinically-asymptomatic (n = 116), seronegative/cardiac subjects (n = 102), and seronegative/healthy subjects (n = 45) were analyzed for peripheral blood biomarkers. RESULTS: Seropositive/chagasic subjects exhibited an increase in sera or plasma levels of myeloperoxidase (MPO, 2.8-fold), advanced oxidation protein products (AOPP, 56%), nitrite (5.7-fold), lipid peroxides (LPO, 12-17-fold) and malondialdehyde (MDA, 4-6-fold); and a decline in superoxide dismutase (SOD, 52%) and glutathione (GSH, 75%) contents. Correlation analysis identified a significant (p<0.001) linear relationship between inflammatory markers (AOPP/nitrite: r = 0.877), inflammation and antioxidant/oxidant status (AOPP/glutathione peroxidase (GPX): r = 0.902, AOPP/GSH: r = 0.806, Nitrite/GPX: 0.773, Nitrite/LPO: 0.805, MDA/MPO: 0.718), and antioxidant/oxidant levels (GPX/MDA: r = 0.768) in chagasic subjects. Of these, MPO, LPO and nitrite biomarkers were highly specific and sensitive for distinguishing seropositive/chagasic subjects from seronegative/healthy controls (p<0.001, training and fitting AUC/ROC >0.95). The MPO (r = 0.664) and LPO (r = 0.841) levels were also correlated with clinical disease state in chagasic subjects (p<0.001). Seronegative/cardiac subjects exhibited up to 77% decline in SOD, 3-5-fold increase in LPO and glutamate pyruvate transaminase (GPT) levels, and statistically insignificant change in MPO, AOPP, MDA, GPX, GSH, and creatine kinase (CK) levels. CONCLUSIONS: The interlinked effects of innate immune responses and antioxidant/oxidant imbalance are major determinants of human Chagas disease. The MPO, LPO and nitrite are excellent biomarkers for diagnosing seropositive/chagasic subjects, and MPO and LPO levels have potential utility in identifying clinical severity of Chagas disease.

Antigenicity and diagnostic potential of vaccine candidates in human Chagas disease.

BACKGROUND: Chagas disease, caused by Trypanosoma cruzi, is endemic in Latin America and an emerging infectious disease in the US and Europe. We have shown TcG1, TcG2, and TcG4 antigens elicit protective immunity to T. cruzi in mice and dogs. Herein, we investigated antigenicity of the recombinant proteins in humans to determine their potential utility for the development of next generation diagnostics for screening of T. cruzi infection and Chagas disease. METHODS AND RESULTS: Sera samples from inhabitants of the endemic areas of Argentina-Bolivia and Mexico-Guatemala were analyzed in 1(st)-phase for anti-T. cruzi antibody response by traditional serology tests; and in 2(nd)-phase for antibody response to the recombinant antigens (individually or mixed) by an ELISA. We noted similar antibody response to candidate antigens in sera samples from inhabitants of Argentina and Mexico (n=175). The IgG antibodies to TcG1, TcG2, and TcG4 (individually) and TcG(mix) were present in 62-71%, 65-78% and 72-82%, and 89-93% of the subjects, respectively, identified to be seropositive by traditional serology. Recombinant TcG1- (93.6%), TcG2- (96%), TcG4- (94.6%) and TcG(mix)- (98%) based ELISA exhibited significantly higher specificity compared to that noted for T. cruzi trypomastigote-based ELISA (77.8%) in diagnosing T. cruzi-infection and avoiding cross-reactivity to Leishmania spp. No significant correlation was noted in the sera levels of antibody response and clinical severity of Chagas disease in seropositive subjects. CONCLUSIONS: Three candidate antigens were recognized by antibody response in chagasic patients from two distinct study sites and expressed in diverse strains of the circulating parasites. A multiplex ELISA detecting antibody response to three antigens was highly sensitive and specific in diagnosing T. cruzi infection in humans, suggesting that a diagnostic kit based on TcG1, TcG2 and TcG4 recombinant proteins will be useful in diverse situations.

Tatividade física como adjunto terapêutico para pacientes psiquiátricos com adoecimento mental severo: revisão da literatura / Physical activity as a therapeutic adjunct to psychiatric patients with severe mental illness: a literature review / Actividad física como coadyuvante terapéutico para pacientes psiquiátricos con enfermedad mental severa: revisión de la literatura

Existe a carência de esclarecimentos mais precisos sobre as repercussões psíquicas da prática de atividade física (AF) nos serviços de saúde mental para pessoas com adoecimento mental severo (severe mental illness; SMI). O objetivo desta revisão foi investigar a AF como adjunto terapêutico para pacientes com SMI. Foram usadas as bases de dados EBSCOHost, PsycInfo, SportDiscus, Medline, PsyArticles. Estudos qualitativos e quantitativos publicados entre 2000 e 2014 foram inclusos e também literatura anterior considerada relevante. As investigações relatam que a AF pode contribuir para a melhora de humor, estado de alerta, concentração, padrões de sono e sintomas psicóticos; aliviar sintomas secundários como baixa autoestima e retraimento social. Há evidência também de benefícios psicológicos da AF em pacientes depressivos. Os pacientes veem a AF positivamente e gostariam de ser ativos e melhorar a saúde mental. Barreiras identificadas pelos pacientes foram: impacto do adoecimento e efeitos de medicação, fadiga, ganho de peso por medicação, medo de discriminação e questões relativas à segurança e falta de suporte profissional. O contexto grupal aparece como um ambiente favorecedor da prática de AF. A literatura considera a AF como uma intervenção que pode ser útil para a promoção da saúde mental e bem-estar, portanto uma estratégia em reabilitação psicossocial para pessoas com SMI. As conclusões dos estudos se restringem a um âmbito descritivo, são limitadas, carecem de uma metodologia de pesquisa precisa em alguns casos, revelando a necessidade de novos estudos.

There is a lack of more precise clarification of the psychological impact of physical activity (PA) in Mental Health Services for people with severe mental illness (SMI). This review aims to investigate physical activity as a therapeutic adjunct for patients with SMI. Data sources: EBSCOHost, PsycInfo, SPORTDiscus, Medline, PsyArticles. Quantitative and qualitative articles published between 2000-2014 were sourced. The studies report that PA can contribute to improve mood, alertness, concentration, sleep patterns and psychotic symptoms. It can relieve secondary symptoms such as low self-esteem and social withdrawal. There is also evidence of psychological benefits of PA in depressed patients. Patients see PA positively and would like to be active and improve mental health. Barriers to uptake of PA were identified: impact of the illness and effects of medication, fatigue, weight gain by medication, fear of discrimination and safety issues and lack of professional support. The group context appears as an environment that promotes the practice of PA. The literature considers AP as an intervention that can be useful for promoting mental health, well-being, therefore a psychosocial rehabilitation strategy for people with SMI. The conclusions of the studies are restricted to a descriptive level, are limited, and lack an accurate methodology of research in some cases revealing the need for future studies.

Existe la carencia de aclaraciones más precisas sobre las repercusiones psíquicas de la práctica de actividad física (AF) en los Servicios de Salud Mental para personas con enfermedad mental severa (severe mental illnes; SMI). El objetivo de esta revisión fue investigar la AF como coadyuvante terapéutico para pacientes con SMI. Fueron usadas las bases de datos EBSCOHost, PsycInfo, SportDiscus, Medline, PsyArticles. Se incluyeron estudios cualitativos y cuantitativos publicados en el período entre 2000- 2014 y la literatura anterior considerada relevante. Los estudios relatan que la AF puede contribuir a la mejora del estado de ánimo, estado de alerta, concentración, patrones de sueño y síntomas psicóticos; aliviar los síntomas secundarios como baja autoestima y retraimiento social. También hay evidencia de beneficios psicológicos de la AF en pacientes depresivos. Los pacientes ven la AF de forma positiva y desean ser activos y mejorar la salud mental. Las barreras identificadas por los pacientes fueron: impacto de la enfermedad y efectos de la medicación, fatiga, aumento de peso por medicación, miedo a la discriminación y cuestiones relativas a la seguridad y falta de soporte profesional. El contexto grupal aparece como un entorno favorecedor de la práctica de AF. La literatura considera la AF como una intervención que puede ser útil para la promoción de la salud mental y el bienestar, por lo tanto una estrategia en rehabilitación psicosocial para las personas con SMI. Las conclusiones de los estudios se restringen a un ámbito descriptivo, son limitadas, carecen de una metodología de investigación precisa en algunos casos revelando la necesidad de nuevos estudios.