Resumo Fundamento A intervenção percutânea em pacientes com disfunção de prótese biológica mitral apresenta-se como uma alternativa ao tratamento cirúrgico convencional. Objetivo Relatar a primeira experiência brasileira de implante transcateter de bioprótese valve-in-valve mitral via transeptal (TMVIV-via transeptal). Métodos Foram incluídos pacientes portadores de disfunção de bioprótese cirúrgica submetidos ao TMVIV-transeptal em 12 hospitais brasileiros. Foram considerados estatisticamente significativos valores de p<0,05. Resultados Entre junho/2016 e fevereiro/2019, 17 pacientes foram submetidos ao TMVIV-via transeptal. A mediana de idade foi 77 anos (IIQ,70-82), a mediana do escore STS-PROM foi 8,7% (IIQ,7,2-17,8). Todos os pacientes tinham sintomas limitantes de insuficiência cardíaca (CF≥III), tendo 5 (29,4%) sido submetidos a mais de uma toracotomia prévia. Obteve-se sucesso do TMVIV-via transeptal em todos os pacientes. A avaliação ecocardiográfica demonstrou redução significativa do gradiente médio (pré-intervenção, 12±3,8 mmHg; pós-intervenção, 5,3±2,6 mmHg; p<0,001), assim como aumento da área valvar mitral (pré-intervenção, 1,06±0,59 cm2; pós-intervenção, 2,18±0,36 cm2; p<0,001) sustentados em 30 dias. Houve redução significativa e imediata da pressão sistólica de artéria pulmonar, com redução adicional em 30 dias (pré-intervenção, 68,9±16,4 mmHg; pós-intervenção, 57,7±16,5 mmHg; 30 dias, 50,9±18,7 mmHg; p<0,001). Durante o seguimento, com mediana de 162 dias (IIQ, 102-411), observou-se marcada melhora clínica (CF≤II) em 87,5%. Um paciente (5,9%) apresentou obstrução de via de saída de ventrículo esquerdo (VSVE), evoluindo para óbito logo após o procedimento, e outro morreu aos 161 dias de seguimento.Conclusão: A primeira experiência brasileira de TMVIV-transeptal demonstra a segurança e a efetividade dessa nova técnica. A obstrução da VSVE é uma complicação potencialmente fatal, reforçando a importância da seleção dos pacientes e do planejamento do procedimento. (Arq Bras Cardiol. 2020; [online].ahead print, PP.0-0)

Abstract Background Percutaneous intervention in patients with bioprosthetic mitral valve dysfunction is an alternative to conventional surgical treatment. Objectives To report the first Brazilian experience with transseptal transcatheter bioprosthetic mitral valve-in-valve implantation (transseptal-TMVIV). Methods Patients with surgical bioprosthetic dysfunction submitted to transseptal-TMVIV in 12 Brazilian hospitals were included. The significance level adopted was p<0.05. Results From June/2016 to February/2019, 17 patients underwent transseptal-TMVIV. Their median age was 77 years (IQR,70-82) and median Society of Thoracic Surgeons predicted risk of mortality (STS-PROM) score was 8.7% (IQR,7.2-17.8). All patients had limiting symptoms of heart failure (FC≥III) and 5 (29.4%) had undergone more than one previous thoracotomy. Transseptal-TMVIV was successful in all patients. Echocardiographic assessment showed a significant reduction in mean mitral valve gradient (pre-intervention, 12±3.8 mmHg; post-intervention, 5.3±2.6 mmHg; p<0.001), in addition to an increase in mitral valve area (pre-intervention, 1.06±0.59 cm2; post-intervention, 2.18±0.36 cm2; p<0.001) sustained for 30 days. There was a significant and immediate reduction in the pulmonary artery systolic pressure, with an additional reduction in 30 days (pre-intervention, 68.9±16.4 mmHg; post-intervention, 57.7±16.5 mmHg; 30 days, 50.9±18.7 mmHg; p<0.001). During follow-up (median, 162 days; IQR, 102-411), significant clinical improvement (FC≤II) was observed in 87.5% of the patients. One patient (5.9%) had left ventricular outflow tract (LVOT) obstruction and died right after the procedure, and another died at 161 days of follow-up. Conclusion The first Brazilian experience with transseptal-TMVIV shows the safety and effectivity of the new technique. The LVOT obstruction is a potentially fatal complication, reinforcing the importance of patients’ selection and of procedural planning. (Arq Bras Cardiol. 2020; [online].ahead print, PP.0-0)

Dynamic S-palmitoylation of proteins is the addition of palmitic acid by zDHHC palmitoyl transferases (PATs) and depalmitoylation by palmitoyl protein thioesterases (PPTs). A putative PAT (TcPAT1) has been previously identified in Trypanosoma cruzi, the etiological agent of Chagas disease. Here we analyse other 14 putative TcPATs and 2 PPTs in the parasite genome. T. cruzi cell lines expressing TcPATs and TcPPTs plus a FLAG tag at the C terminus were produced for most enzymes, with positive detection by indirect immunofluorescence. Overexpressed TcPATs were mostly found as single spots at the parasite anterior end, while the TcPPTs were dispersed throughout the parasite body.

BACKGROUND Recent studies showed that essential oils from different pepper species (Piper spp.) have promising leishmanicidal and trypanocidal activities. OBJECTIVES In search for natural compounds against Trypanosoma cruzi, different forms of the parasite were incubated for 24 h at 28ºC or 4ºC with Piper aduncum essential oil (PaEO) or its main constituents linalool and nerolidol. METHODS PaEO chemical composition was obtained by GC-MS. Drug activity assays were based on cell counting, MTT data or infection index values. The effect of PaEO on the T. cruzi cell cycle and mitochondrial membrane potential was evaluated by flow cytometry. FINDINGS PaEO was effective against cell-derived (IC50/24 h: 2.8 μg/mL) and metacyclic (IC50/24 h: 12.1 μg/mL) trypomastigotes, as well as intracellular amastigotes (IC50/24 h: 9 μg/mL). At 4ºC - the temperature of red blood cells (RBCs) storage in blood banks - cell-derived trypomastigotes were more sensitive to PaEO (IC50/24 h = 3.8 μg/mL) than to gentian violet (IC50/24 h = 24.7 mg/mL). Cytotoxicity assays using Vero cells (37ºC) and RBCs (4ºC) showed that PaEO has increased selectivity for cell-derived trypomastigotes. Flow cytometry analysis showed that PaEO does not affect the cell cycle of T. cruzi epimastigotes, but decreases their mitochondrial membrane potential. GC-MS data identified nerolidol and linalool as major components of PaEO, and linalool had trypanocidal effect (IC50/24 h: 306 ng/mL) at 4ºC. MAIN CONCLUSION The trypanocidal effect of PaEO is likely due to the presence of linalool, which may represent an interesting candidate for use in the treatment of potentially contaminated RBCs bags at low temperature.

Resumo Objetivo: analisar a percepção sobre a atividade de trabalho dos agentes de limpeza terceirizados de uma faculdade pública e determinar seu índice de capacidade para o trabalho. Métodos: foram utilizadas três ferramentas - Análise Coletiva do Trabalho (ACT); Questionário Sociodemográfico de trabalho e de estilo de vida (QSD) e o Índice de Capacidade para o Trabalho (ICT). Resultados/discussão: das 22 agentes de limpeza, seis apresentaram ICT Baixo e Moderado, o que as coloca em posição de vulnerabilidade. Para além das demandas físicas de trabalho, as queixas das agentes de limpeza se concentraram nos riscos psicossociais, especialmente relacionados ao estresse e ao assédio moral. As trabalhadoras acreditam no valor de suas atividades, mas manifestaram ressentimento pela falta de reconhecimento por parte dos gestores e usuários da instituição. Conclusão: Fica evidente a necessidade de intervir nesse contexto para a manutenção da capacidade de trabalho e das condições para viver bem destes agentes de limpeza terceirizados.

Abstract Objective: to analyze the perception of the outsourced cleaning workers from a public university on their work activity and to determine their work ability index. Method: three tools were used - Collective Work Analysis; a social-demographic, occupational and lifestyle questionnaire; and Work Ability Index (WAI). Results/discussion: from the 22 cleaning workers, six presented low and moderated WAI, which shows that they are in a vulnerable position. Besides the physical demands, the workers’ complaints were about psychosocial risks, specially related to stress and workplace bullying. Discussion: The cleaning workers believe in the quality value that their activities add to the university and expressed resentment by the lack of recognition from the managers and users of the institution. It is clear that the university needs to intervene in those work conditions so that workers can maintain their work capacity and well being.

This study had analyzed the antibacterial, antifungal and trypanocidal activity of the essential oils from Cinnamodendron dinisii Schwacke (Canellaceae) and Siparuna guianensis Aublet (Siparunaceae). The essential oils were obtained from fresh leaves by hydrodistillation, using a modified Clevenger apparatus. Chemical analysis by gas-liquid chromatography coupled to mass spectrometry (GC-MS) showed that these essential oils are rich in monoterpene and sesquiterpene hydrocarbons. Activity against the pathogenic bacteria Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella choleraesuis and Staphylococcus aureus was evaluated with the agar cavity diffusion method, while activity on the filamentous fungi Aspergillus flavus, Aspergillus niger, Aspergillus carbonariusand Penicillium commune was evaluated by the disk diffusion technique. Trypanocidal activity was tested against Trypanosoma cruzi epimastigotes, using the Tetrazolium salt (MTT) colorimetric assay. Both essential oils exhibited low inhibitory effect towards bacteria, showing high MIC values (125–500 μg mL−1), with Gram positive bacteria being more susceptible. Better inhibitory effect was obtained for the evaluated fungi, with lower MIC values (7.81–250 μg mL−1), being A. flavus the most susceptible species. Both essential oils presented low trypanocidal activity, with IC50/24 h values of 209.30 μg mL−1 for S. guianensis and 282.93 μg mL−1 for C. dinisii. Thus, the high values observed for the MIC of evaluated bacteria and for IC50/24 h of T. cruzi, suggest that the essential oils have a low inhibitory activity against these microorganisms. In addition, the low MIC values observed for the tested fungi species indicate good inhibitory activity on these microorganisms’s growth.

Heteroxenic and monoxenic trypanosomatids were screened for the presence of actin using a mouse polyclonal antibody produced against the entire sequence of the Trypanosoma cruzi actin gene, encoding a 41.9 kDa protein. Western blot analysis showed that this antibody reacted with a polypeptide of approximately 42 kDa in the whole-cell lysates of parasites targeting mammals (T. cruzi, Trypanosoma brucei and Leishmania major), insects (Angomonas deanei, Crithidia fasciculata, Herpetomonas samuelpessoai and Strigomonas culicis) and plants (Phytomonas serpens). A single polypeptide of approximately 42 kDa was detected in the whole-cell lysates of T. cruzi cultured epimastigotes, metacyclic trypomastigotes and amastigotes at similar protein expression levels. Confocal microscopy showed that actin was expressed throughout the cytoplasm of all the tested trypanosomatids. These data demonstrate that actin expression is widespread in trypanosomatids.

Trypanosomes are parasitic protozoa in which gene expression is primarily controlled through the regulation of mRNA stability and translation. This post-transcriptional control is mediated by various families of RNA-binding proteins, including those with zinc finger CCCH motifs. CCCH zinc finger proteins have been shown to be essential to differentiation events in trypanosomatid parasites. Here, we functionally characterise TcZFP2 as a predicted post-transcriptional regulator of differentiation in Trypanosoma cruzi. This protein was detected in cell culture-derived amastigotes and trypomastigotes, but it was present in smaller amounts in metacyclic trypomastigote forms of T. cruzi. We use an optimised recombinant RNA immunopreciptation followed by microarray analysis assay to identify TcZFP2 target mRNAs. We further demonstrate that TcZFP2 binds an A-rich sequence in which the adenosine residue repeats are essential for high-affinity recognition. An analysis of the expression profiles of the genes encoding the TcZFP2-associated mRNAs throughout the parasite life cycle by microarray hybridisation showed that most of the associated mRNAs were upregulated in the metacyclic trypomastigote forms, also suggesting a role for TcZFP2 in metacyclic trypomastigote differentiation. Knockdown of the orthologous Trypanosoma brucei protein levels showed ZFP2 to be a positive regulator of specific target mRNA abundance.

The family Nematotaeniidae, tapeworms commonly found in the small intestines of amphibians and reptiles, includes 27 recognised species distributed among four genera: Bitegmen Jones, Cylindrotaenia Jewell, Distoichometra Dickey and Nematotaenia Lühe. The taxonomy of these cestodes is poorly defined, due in part to the difficulties of observing many anatomical traits. This study presents and describes a new genus and species of nematotaeniid parasite found in cane toads (Rhinella marina) from eastern Brazilian Amazonia. The cestodes were collected during the necropsy of 20 hosts captured in the urban area of Belém, Pará. The specimens were fixed and processed for light microscopy, scanning electron microscopy (SEM) and three-dimensional (3D) reconstruction. Samples were also collected for molecular analyses. The specimens presented a cylindrical body, two testes and paruterine organs. However, they could not be allocated to any of the four existing nematotaeniid genera due to the presence of two each of dorsal compact medullary testes, cirri, cirrus pouches, genital pores, ovaries and vitelline glands per mature segment. Lanfrediella amphicirrus gen. nov. sp. nov. is the first nematotaeniid studied using Historesin analysis, SEM and 3D reconstruction, and it is the second taxon for which molecular data have been deposited in GenBank.

Citral, the main constituent of lemongrass (Cymbopogon citratus) essential oil, was added to Trypanosoma cruzi cultures grown in TAU3AAG medium to observe the effect on the epimastigote-to-trypomastigote differentiation process (metacyclogenesis). Our results showed that citral (20 μg/mL) did not affect epimastigote viability or inhibit the differentiation process. Concentrations higher than 60 μg/mL, however, led to 100% cell death (both epimastigote and trypomastigote forms). Although epimastigotes incubated with 30 μg/mL citral were viable and able to adhere to the substrate, we observed around 50% inhibition in metacyclogenesis, with a calculated concentration that inhibited metacyclogenesis by 50% after 24 h (IC50/24 h) of about 31 μg/mL. Treatment with 30 μg/mL citral did not hinder epimastigote multiplication because epimastigote growth resumed when treated cells were transferred to a drug-free liver infusion tryptose culture medium. Metacyclogenesis was almost totally abolished at 40 μg/mL after 24 h of incubation. Furthermore, the metacyclic trypomastigotes obtained in vitro were similarly susceptible to citral, with an IC50/24 h, concentration that killed 50% of the cells after 24 h, of about 24.5 μg/mL. Therefore, citral appears to be a good candidate as an inhibitory drug for further studies analyzing the T. cruzi metacyclogenesis process.

O objetivo desta pesquisa foi traçar o perfil demográfico e práticas de saúde bucal do ACS, em cidades de pequeno porte do Estado do Piauí. Foram selecionados quatro municípios: Água Branca, Piracuruca, Queimada Nova e Simões. O método utilizado foi o transversal observacional descritivo. Foram aplicados questionários a 109 ACS, constituídos de 28 perguntas fechadas. Os dados foram transferidos para planilha do programa ExcelÒ, tabulados e analisados. Pelos dados obtidos, foi possível concluir que: (1) o agente comunitário de saúde do interior do Piauí é predominantemente do sexo feminino, casado, com idade entre 20-39 anos, tem em média de um a três filhos, com grau de escolaridade médio acima de nove anos, renda de um salário mínimo, residindo em média há 24 anos na comunidade; (2) a grande maioria dos ACS não foi capacitada, não assistiu palestras educativas sobre saúde bucal; entretanto, quase a metade realiza atividades em saúde bucal, mas não as registra; (3) a autopercepção sobre o conhecimento em saúde bucal pelos ACS predominou entre conceitos regular e bom, o que coincidiu com o percentual de acertos ao questionário aplicado.

The aim of this survey was to draw the demographic profile and the actions concerning oral health carried out by the communitarian agents of health, in small towns of Piauí State, Brazil. Four towns were chosen: Água Branca, Piracuruca, Queimada Nova e Simões. The method used was observational descriptive cross-sectional. One hundred and nine agents were assessed through questionnaires, with 28 multiples choice questions. Data was analyzed in Excel. It was possible to conclude that: the communitarian agents on the countryside of Piauí are predominantly females, married, age between 20-39, with one to three children, nine years of formal education, monthly income of one minimum salary, and has been living in their community for 24 years in average; the large majority of the agents has not received specific qualification to carry out their job, has not attended lectures on oral health, although almost half of them works on activities related to oral health, despite not registering them properly; they perceive themselves as having a fair knowledge of oral health, which coincided with the assessment collected by the questionnaire.

O objetivo desta pesquisa foi traçar o perfil demográfico e práticas de saúde bucal do ACS, em cidades de pequeno porte do Estado do Piauí. Foram selecionados quatro municípios: Água Branca, Piracuruca, Queimada Nova e Simões. O método utilizado foi o transversal observacional descritivo. Foram aplicados questionários a 109 ACS, constituídos de 28 perguntas fechadas. Os dados foram transferidos para planilha do programa ExcelÒ, tabulados e analisados. Pelos dados obtidos, foi possível concluir que: (1) o agente comunitário de saúde do interior do Piauí é predominantemente do sexo feminino, casado, com idade entre 20-39 anos, tem em média de um a três filhos, com grau de escolaridade médio acima de nove anos, renda de um salário mínimo, residindo em média há 24 anos na comunidade; (2) a grande maioria dos ACS não foi capacitada, não assistiu palestras educativas sobre saúde bucal; entretanto, quase a metade realiza atividades em saúde bucal, mas não as registra; (3) a autopercepção sobre o conhecimento em saúde bucal pelos ACS predominou entre conceitos regular e bom, o que coincidiu com o percentual de acertos ao questionário aplicado.

The aim of this survey was to draw the demographic profile and the actions concerning oral health carried out by the communitarian agents of health, in small towns of Piauí State, Brazil. Four towns were chosen: Água Branca, Piracuruca, Queimada Nova e Simões. The method used was observational descriptive cross-sectional. One hundred and nine agents were assessed through questionnaires, with 28 multiples choice questions. Data was analyzed in Excel. It was possible to conclude that: the communitarian agents on the countryside of Piauí are predominantly females, married, age between 20-39, with one to three children, nine years of formal education, monthly income of one minimum salary, and has been living in their community for 24 years in average; the large majority of the agents has not received specific qualification to carry out their job, has not attended lectures on oral health, although almost half of them works on activities related to oral health, despite not registering them properly; they perceive themselves as having a fair knowledge of oral health, which coincided with the assessment collected by the questionnaire.

Uptake of transferrin by epimastigote forms of the protozoan Trypanosoma cruzi occurs mainly through a cytostome/ cytopharynx, via uncoated endocytic vesicles that bud off from the bottom of the cytopharynx. We have here examined whether detergent-resistant membrane (DRM) domains might be involved in this process. Purified whole cell membrane fractions were assayed for cholesterol levels and used in dot blot analyses. Detergent-resistant membrane markers (cholera B toxin and anti-flotillin-1 antibody) presented positive reaction by dot blots in cholesterol-rich/ protein-poor membrane sub-fractions. The positive dot blot fraction was submitted to lipid composition analysis, showing composition similar to that of raft fractions described for other eukaryotic cells. Immunofluorescence assays allowed the localization of punctual positive signal for flotillin-1, matching the precise cytostome/ cytopharynx location. These data were confirmed by immunofluorescence assays with the co-localization of flotillin-1 and the transferrin uptake site. Our data suggest that DRM domains occur and are integrated at the cytostome/ cytopharynx of T. cruzi epimastigotes, being the main route for transferrin uptake.

Experimental chronic (45-day-old) skin lesion in hamster hind foot induced by Leishmania (Viannia) lainsoni infection showed the presence of promastigote forms in the tissue, inside parasitophorous vacuoles, as assessed by transmission electron microscopy. Experimental in vitro interaction (24 and 48 h) between Leishmania (V.)lainsoni and J774-G8 macrophage cells also demonstrated the same profile. This morphological aspect is unusual, since in this parasite genus only amastigote forms have been described as the resistant and obligate intracellular forms.

Este trabalho objetivou identificar locos de carateres quantitativos (QTLs) assosciados com fotossíntese, crescimento e produtividade de soja com a finalidade de identificar possíveis associações entre estas características pela análise de coicindência de QTLs nos respectivos grupos de ligação (GL). Para isto, foram usadas populações de linhagens recombinantes endogâmicas (RILs) das gerações F7 e F8 derivadas dos cruzamentos entre as variedades BARC-8 e Garimpo. As características avaliadas foram taxa de assimilação líquida de CO2 sob luz saturante (Asat), taxa de fotossíntese potencial (Pmax), área foliar (A), área foliar específica (SLA), nitrogênio foliar específico (N); número de nódulos (nN), sementes (n s) e vagens (nP); massa fresca de sementes por planta (W S), massa fresca de 100 sementes (W HS) e percentagem de proteína da semente (P%). Foi possível identificar os seguintes QTLs associados com as características: SLA, Asat, N, W R, W ST, W L, W T, W P, W HS, n s e nP, indicando que as populações de RIL apresentam grande potencial para mapeamento de locos associados com caracteres quantitativos em soja. As correlações entre os caracteres analisados foram parcialmente confirmadas pela coincidência de QTL.

This work aimed to identify the quantitative trait loci (QTL) associated with photosynthesis and growth and productivity traits of soybean and to study possible associations between these traits by the analysis of coincidence of QTL in linkage groups (LGs). Thus, populations of recombinant inbred lines (RILs) of the F7 and F8 generations derived from the cross between the varieties BARC-8 and Garimpo were used. The traits evaluated were net assimilation rate of CO2 under saturating light (Asat), potential photosynthesis rate (Pmax), leaf area (A), specific leaf area (SLA), specific leaf nitrogen (N); root (W R), nodule (W N), stem (W ST), leaf (W L), pod (W P) and plant dry mass (W T); nodule (nN), seed (n s), and pod number (nP); seed fresh mass per plant (W S), one-hundred seed fresh mass (W HS) and seed protein percentage (P%). It was possible to identify the following QTL associated with the following soybean traits: SLA, Asat, N, W R, W ST, W L, W T, W P, W HS, n s and nP, indicating that the RIL population has a great potential for mapping loci associated with quantitative traits of the soybean crop. The correlations between the soybean traits were partially confirmed by coincidence of QTL.

Leishmania (Viannia) lainsoni is the Leishmania species that presents the most distinct biological (morphology, growth in axenic culture medium), biochemical (enzymatic electrophoresis profile), and molecular biology characteristics, when compared to other species of the Viannia subgenus. Development of promastigote forms of this parasite attached to the wall of the pyloric and hind gut regions of sand fly vectors is a solid characteristic that allows its positioning in the Viannia subgenus. However, taxonomic data from biochemical and molecular techniques on this Leishmania species are still not conclusive. It is evident the difficulty in taxonomically positioning this borderline Leishmania species. In this review we present the data accumulated since L. (Viannia) lainsoni has been described and we discuss its position in the Viannia subgenus.