Expansion of the cytochrome C oxidase subunit I database and description of four new lobose testate amoebae species (Amoebozoa; Arcellinida).

Arcellinida is ascending in importance in protistology, but description of their diversity still presents multiple challenges. Furthermore, applicable tools for surveillance of these organisms are still in developing stages. Importantly, a good database that sets a correspondence between molecular barcodes and species morphology is lacking. Cytochrome oxidase (COI) has been suggested as the most relevant marker for species discrimination in Arcellinida. However, some major groups of Arcellinida are still lacking a COI sequence. Here we expand the database of COI marker sequences for Arcellinids, using single-cell PCR, transcriptomics, and database scavenging. In the present work, we added 24 new Arcellinida COI sequences to the database, covering all unsampled infra- and suborders. Additionally, we added six new SSUrRNA sequences and described four new species using morphological, morphometrical, and molecular evidence: Heleopera steppica, Centropyxis blatta, Arcella uspiensis, and Cylindrifflugia periurbana. This new database will provide a new starting point to address new research questions from shell evolution, biogeography, and systematics of arcellinids.

A decade of Zoology Summer Course: impressions and impacts of the first university extension course on Zoology in Brazil / Uma década de Curso de Verão em Zoologia: impressões e impactos do primeiro curso de extensão universitária sobre Zoologia no Brasil

Although the diversity of animal groups distributed in Brazil provides countless research opportunities, the current scenario does not follow this demand. The reasons for the disconnections range from inequality in the availability of resources for teaching and research to the focus of researchers on specific groups of animals, while others remain neglected. Training new potential Brazilian researchers interested in Zoology is essential for a greater understanding of this diversity, as well as exposing those potential new researchers to new groups and different work possibilities. Thus, the Summer Course in Zoology (in Portuguese, CVZoo) promoted by the Graduate Program in Zoology at the University of São Paulo, over the last ten years, has been seeking to contribute to this training of new researchers in the field of Zoology, as well as in updating teachers through university extension activities. In order to assess the impacts caused by CVZoo on the academic and professional training of the participants, Google forms were sent to participants in the ten editions of the course, as well as compiled information available on the Lattes Platform. Qualitative and quantitative analyses showed the profile of graduates, their expectations, and perceptions about the course. Based on these data, we demonstrate the CVZoo's efficiency in popularizing Zoology throughout the country in contributing to the decentralization of knowledge as well as in meeting the urgent concerns of making access to knowledge more egalitarian and socially fair.

Embora a diversidade de grupos de animais existentes no Brasil ofereça inúmeras oportunidades de estudo, o cenário atual não acompanha essa demanda. Os motivos para essa desconexão vão desde a desigualdade na disponibilidade de recursos para ensino e pesquisa até o foco de pesquisadores em grupos específicos de animais, enquanto outros permanecem negligenciados. O treinamento de novos pesquisadores interessados em Zoologia é essencial para um maior entendimento da diversidade brasileira, assim como a exposição de tais pesquisadores a novos grupos e diferentes possibilidades de trabalho. O Curso de Verão em Zoologia (CVZoo) promovido pelo Programa de Pós-graduação em Zoologia da Universidade de São Paulo, ao longo de dez anos vem buscando contribuir para a formação de novos(as) pesquisadores(as) na área da Zoologia, bem como na atualização de docentes do Ensino Básico por meio de atividades de extensão universitária. Para avaliar os impactos causados pelo CVZoo na formação acadêmica e profissionalizante dos participantes, foram enviados formulários aos participantes das dez edições do curso, bem como compiladas informações disponíveis na Plataforma Lattes. Análises qualitativas e quantitativas evidenciaram o perfil das pessoas egressas, suas expectativas e percepções acerca do curso oferecido. Com base nesses dados, é apontada a eficiência do CVZoo na popularização da Zoologia por todo o país, contribuindo para a descentralização do conhecimento, bem como atendendo às preocupações prementes de tornar o acesso ao conhecimento mais igualitário e socialmente justo

Deconstructing Difflugia: The tangled evolution of lobose testate amoebae shells (Amoebozoa: Arcellinida) illustrates the importance of convergent evolution in protist phylogeny.

Protists, the micro-eukaryotes that are neither plants, animals nor fungi build up the greatest part of eukaryotic diversity on Earth. Yet, their evolutionary histories and patterns are still mostly ignored, and their complexity overlooked. Protists are often assumed to keep stable morphologies for long periods of time (morphological stasis). In this work, we test this paradigm taking Arcellinida testate amoebae as a model. We build a taxon-rich phylogeny based on two mitochondrial (COI and NADH) and one nuclear (SSU) gene, and reconstruct morphological evolution among clades. In addition, we prove the existence of mitochondrial mRNA editing for the COI gene. The trees show a lack of conservatism of shell outlines within the main clades, as well as a widespread occurrence of morphological convergences between far-related taxa. Our results refute, therefore, a widespread morphological stasis, which may be an artefact resulting from low taxon coverage. As a corollary, we also revise the groups systematics, notably by emending the large and highly polyphyletic genus Difflugia. These results lead, amongst others, to the erection of a new infraorder Cylindrothecina, as well as two new genera Cylindrifflugia and Golemanskia.

A comparative study indicates vertical inheritance and horizontal gene transfer of arsenic resistance-related genes in eukaryotes.

Arsenic is a ubiquitous element in the environment, a source of constant evolutionary pressure on organisms. The arsenic resistance machinery is thoroughly described for bacteria. Highly resistant lineages are also common in eukaryotes, but evolutionary knowledge is much more limited. While the origin of the resistance machinery in eukaryotes is loosely attributed to horizontal gene transfer (HGT) from bacteria, only a handful of eukaryotes were deeply studied. Here we investigate the origin and evolution of the core genes in arsenic resistance in eukaryotes using a broad phylogenetic framework. We hypothesize that, as arsenic pressure is constant throughout Earth's history, resistance mechanisms are probably ancestral to eukaryotes. We identified homologs for each of the arsenic resistance genes in eukaryotes and traced their possible origin using phylogenetic reconstruction. We reveal that: i. an important component of the arsenic-resistant machinery originated before the last eukaryotic common ancestor; ii. later events of gene duplication and HGT generated new homologs that, in many cases, replaced ancestral ones. Even though HGT has an important contribution to the expansion of arsenic metabolism in eukaryotes, we propose the hypothesis of ancestral origin and differential retention of arsenic resistance mechanisms in the group. Key-words: Environmental adaptation; resistance to toxic metalloids; detoxification; comparative genomics; functional phylogenomics.

Analyses of the pericyte transcriptome in ischemic skeletal muscles.

BACKGROUND: Peripheral arterial disease (PAD) affects millions of people and compromises quality of life. Critical limb ischemia (CLI), which is the most advanced stage of PAD, can cause nonhealing ulcers and strong chronic pain, and it shortens the patients' life expectancy. Cell-based angiogenic therapies are becoming a real therapeutic approach to treat CLI. Pericytes are cells that surround vascular endothelial cells to reinforce vessel integrity and regulate local blood pressure and metabolism. In the past decade, researchers also found that pericytes may function as stem or progenitor cells in the body, showing the potential to differentiate into several cell types. We investigated the gene expression profiles of pericytes during the early stages of limb ischemia, as well as the alterations in pericyte subpopulations to better understand the behavior of pericytes under ischemic conditions. METHODS: In this study, we used a hindlimb ischemia model to mimic CLI in C57/BL6 mice and explore the role of pericytes in regeneration. To this end, muscle pericytes were isolated at different time points after the induction of ischemia. The phenotypes and transcriptomic profiles of the pericytes isolated at these discrete time points were assessed using flow cytometry and RNA sequencing. RESULTS: Ischemia triggered proliferation and migration and upregulated the expression of myogenesis-related transcripts in pericytes. Furthermore, the transcriptomic analysis also revealed that pericytes induce or upregulate the expression of a number of cytokines with effects on endothelial cells, leukocyte chemoattraction, or the activation of inflammatory cells. CONCLUSIONS: Our findings provide a database that will improve our understanding of skeletal muscle pericyte biology under ischemic conditions, which may be useful for the development of novel pericyte-based cell and gene therapies.

The Sexual Ancestor of all Eukaryotes: A Defense of the "Meiosis Toolkit": A Rigorous Survey Supports the Obligate Link between Meiosis Machinery and Sexual Recombination.

The distribution pattern of the meiotic machinery in known eukaryotes is most parsimoniously explained by the hypothesis that all eukaryotes are ancestrally sexual. However, this assumption is questioned by preliminary results, in culture conditions. These suggested that Acanthamoeba, an organism considered to be largely asexual, constitutively expresses meiosis genes nevertheless-at least in the lab. This apparent disconnect between the "meiosis toolkit" and sexual processes in Acanthamoeba led to the conclusion that the eukaryotic ancestor is asexual. In this review, the "meiosis toolkit" is rigorously defended, drawing on numerous research articles. Additionally, the claim of constitutive meiotic gene expression is probed in Acanthamoeba via the same transcriptomics data. The results show that the expression of the meiotic machinery is not constitutive in Acanthamoeba as claimed before. Furthermore, it is argued that this would have no implications for understanding the nature of the eukaryotic ancestor, regardless of the result.

De novo Sequencing, Assembly, and Annotation of the Transcriptome for the Free-Living Testate Amoeba Arcella intermedia.

Arcella, a diverse understudied genus of testate amoebae is a member of Tubulinea in Amoebozoa group. Transcriptomes are a powerful tool for characterization of these organisms as they are an efficient way of characterizing the protein-coding potential of the genome. In this work, we employed both single-cell and clonal populations transcriptomics to create a reference transcriptome for Arcella. We compared our results with annotations of Dictyostelium discoideum, a model Amoebozoan. We assembled a pool of 38 Arcella intermedia transcriptomes, which after filtering are composed of a total of 14,712 translated proteins. There are GO categories enriched in Arcella including mainly intracellular signal transduction pathways; we also used KEGG to annotate 11,546 contigs, which also have similar distribution to Dictyostelium. A large portion of data is still impossible to assign to a gene family, probably due to a combination of lineage-specific genes, incomplete sequences in the transcriptome and rapidly evolved genes. Some absences in pathways could also be related to low expression of these genes. We provide a reference database for Arcella, and we highlight the emergence of the need for further gene discovery in Arcella.

O processo de trabalho gerencial do enfermeiro no setor de hiperdia na atenção básica: relato de experiência / The nurse's management work process in the hyperdia sector in basic care: experience report

Objetivo: analisar a organização do processo de trabalho gerencial do enfermeiro no setor do Programa de Hiperdia em uma Policlínica Regional do município de Niterói-RJ. Metodologia: trata-se de um relato de experiência, de abordagem qualitativa, desenvolvido a partir da observação da organização do processo de trabalho dos enfermeiros que trabalham no setor do Programa de Hiperdia de uma Policlínica Regional do município de Niterói-RJ. Realizada durante o segundo semestre acadêmico de 2019. Levaram-se em consideração os seguintes componentes do processo: objeto, agentes, instrumentos, finalidade, método e produto. Resultados: foi possível analisar o processo de trabalho exercido pela enfermagem no setor do Programa de Hiperdia. Observou-se a implementação de rotina, utilização de diversos instrumentos, organização de registros de enfermagem e implementação de controle diário de atendimento, cabe ressaltar o bom relacionamento enfermeiro-paciente, que possibilita o cuidado adequado com a necessidade do usuário. Considerações finais: o Programa de Hiperdia se mostra importante para a Policlínica ofertando um atendimento dentro das perspectivas do Sistema Único de Saúde (SUS). No processo de trabalho gerencial o enfermeiro ao coordenar esse setor, realiza função de gestor, pesquisador, educador e prestador de assistência, funções estas que são os pilares construtores do processo de trabalho da enfermagem. (AU)

Objective: to analyze the organization of the managerial work process of nurses in the sector of the Hiperdia Program in a Regional Polyclinic in the city of Niterói-RJ. Methodology: it is an experience report, with a qualitative approach, developed from the observation of the organization of the work process of nurses who work in the sector of the Hiperdia Program of a Regional Polyclinic in the city of Niterói-RJ. Held during the second academic semester of 2019. The following components of the process were taken into account: object, agents, instruments, purpose, method and product. Results: it was possible to analyze the work process performed by nursing in the sector of the Hiperdia Program. It was observed the routine implementation, the use of several instruments, the organization of nursing records and the implementation of daily care control, it is worth mentioning the good nurse-patient relationship, which enables adequate care with the user's need. Final considerations: the Hiperdia Program is important for the Polyclinic, offering care within the perspective of the Unified Health System (SUS). In the managerial work process, the nurse, when coordinating this sector, performs the function of manager, researcher, educator and care provider, functions that are the building blocks of the nursing work process. (AU)

Objetivo: analizar la organización del proceso de trabajo gerencial de las enfermeras en el sector del Programa Hiperdia en un Policlínico Regional en la ciudad de Niterói-RJ. Metodología: este es un informe de experiencia, con un enfoque cualitativo, desarrollado a partir de la observación de la organización del proceso de trabajo de las enfermeras que trabajan en el sector del Programa Hiperdia de una Policlínica Regional en la ciudad de Niterói-RJ. Se llevó a cabo durante el segundo semestre académico de 2019. Se tomaron en cuenta los siguientes componentes del proceso: objeto, agentes, instrumentos, propósito, método y producto. Resultados: fue posible analizar el proceso de trabajo realizado por enfermería en el sector del Programa Hiperdia. Se observó la implementación de rutina, el uso de varios instrumentos, la organización de los registros de enfermería y la implementación del control de atención diaria, vale la pena mencionar la buena relación enfermera-paciente, que permite una atención adecuada a las necesidades del usuario. Consideraciones finales: el Programa Hiperdia es importante para la Policlínica, ya que ofrece atención desde la perspectiva del Sistema Único de Salud (SUS). En el proceso de trabajo gerencial, la enfermera, al coordinar este sector, desempeña la función de gerente, investigador, educador y proveedor de atención, funciones que son los componentes básicos del proceso de trabajo de enfermería. (AU)

Phylogenomics and Morphological Reconstruction of Arcellinida Testate Amoebae Highlight Diversity of Microbial Eukaryotes in the Neoproterozoic.

Life was microbial for the majority of Earth's history, but as very few microbial lineages leave a fossil record, the Precambrian evolution of life remains shrouded in mystery. Shelled (testate) amoebae stand out as an exception with rich documented diversity in the Neoproterozoic as vase-shaped microfossils (VSMs). While there is general consensus that most of these can be attributed to the Arcellinida lineage in Amoebozoa, it is still unclear whether they can be used as key fossils for interpretation of early eukaryotic evolution. Here, we present a well-resolved phylogenomic reconstruction based on 250 genes, obtained using single-cell transcriptomic techniques from a representative selection of 19 Arcellinid testate amoeba taxa. The robust phylogenetic framework enables deeper interpretations of evolution in this lineage and demanded an updated classification of the group. Additionally, we performed reconstruction of ancestral morphologies, yielding hypothetical ancestors remarkably similar to existing Neoproterozoic VSMs. We demonstrate that major lineages of testate amoebae were already diversified before the Sturtian glaciation (720 mya), supporting the hypothesis that massive eukaryotic diversification took place in the early Neoproterozoic and congruent with the interpretation that VSM are arcellinid testate amoebae.

Morphometric and genetic analysis of Arcella intermedia and Arcella intermedia laevis (Amoebozoa, Arcellinida) illuminate phenotypic plasticity in microbial eukaryotes.

Testate amoebae are eukaryotic microorganisms characterized by the presence of an external shell (test). The shell morphology is used as a diagnostic character, but discordance between morphological and molecular data has been demonstrated in groups of arcellinids (Amoebozoa), one of the principal groups of testate amoebae. Morphology of the test is supposed to differentiate genera and species and it is applied in ecological, monitoring and paleontological studies. However, if phenotype does not reflect genotype, conclusions in these types of studies become severely impaired. The objective of this work is to evaluate the morphometrical and morphological variation of the closely related and morphologically similar taxa Arcella intermedia laevis Tsyganov and Mazei, 2006 and Arcella intermedia (Deflandre 1928) Tsyganov and Mazei, 2006 in nature and in cultured individuals and see how these are correlated with molecular data. Our results demonstrate that phenotypic plasticity in Arcella intermedia make morphological distinctions impossible in both taxa. Arcella intermedia and Arcella intermedia laevis are molecularly identical for SSU rDNA and a mitochondrial molecular marker (NAD9/7). We conclude that morphological techniques alone cannot identify phenotypic plasticity from natural populations. More work is clearly needed to better understand the morphological, morphometric and molecular variability in these organisms.