Expression of tumoral GSK3-ß, PD-L1, and CD8 cell density in urothelial carcinomas, association with tumor grade and overall survival.

Bladder cancer is the most common malignancy in the urinary tract, and is biologically and clinically quite heterogeneous. Around 90% of diagnoses are made in the 6th decade, being more prevalent in males. The programmed cell death 1 (PD-1) and programmed cell death ligand 1 (PD-L1) axis play a putative role in immune checkpoint and as a means through which cancer evades the immune system. Inhibition of the glicogênio synthase kinase (GSK) 3 leads to the downregulation of PD-1 via upregulation of the transcription factor Tbet. The use of biomarkers PD-L1 and GSK-3ß and evaluation of the immune infiltrate have very promising correlations with urothelial carcinoma prognosis and treatment prediction. OBJECTIVE: To investigate the protein expression of PD-L1 and GSK-3ß and the CD8-positive immune infiltrates in bladder carcinomas. MATERIALS AND METHODS: This was a cross-sectional study of 140 samples of urothelial carcinomas from 2015 to 2018. Automated digitally assisted scoring and conventional analyses of the markers of GSK-3ß (27C10), CD8 (7103ß) and PDL-1 (22c3), were reviewed by two pathologists independently and a histologic score was calculated. The density of CD8 was also measured. RESULTS: The immunoexpression of GSK-3ß (91%) was presented in most samples, PD-L1 in 62.9% and CD8 cells present in 46.3% of cases. When analyzed in conjunction, the levels of GSK-3ß and PD-L1 (P = 0.033), and CD8 and PD-L1 (P<0.002) showed significant correlations. No significant associations were observed between GSK-3ß and CD8. The positivity of GSK-3ß and PD-L1 was predominant in high-grade tumors. CONCLUSION: Despite the tumor microenvironment heterogeneity, the expression of CD8, GSK-3ß and PDL1 could be valuable and GSK-3ß could be a potential target in advanced bladder cancer, especially in the context of immunotherapy.

Comparison of glass and digital slides for cervical cytopathology screening and interpretation.

Cervical cancer is the second most common form of cancer and a leading cause of premature death among women aged 15 to 44 worldwide. In Brazil, there is a high prevalence of infection by the human papillomavirus - HPV. Digital pathology optimizes time and space for reading cervicovaginal cytology slides. We evaluated the feasibility of using whole slide images (WSI) for the routine interpretation of cytology exams. A total of 99 cases of vaginal cytology were selected from a reference laboratory in Northeastern Brazil. Three cytotechnicians participated in the study. Cellular atypia was the one that most presented concordance values. Two observers almost perfectly agreed (k = 0.86 and k = 0.84, respectively) on the negative diagnoses. The performance of the evaluators for NILM (negative for intraepithelial lesion and malignancy) showed high reproducibility and sensitivity in the digital slides, mainly between evaluators A and C. In contrast, the microbiology group showed disagreement between the diagnoses by digital slides and the standard- gold. The concordance between the digital diagnoses and the gold standard for ASCUS was 89%. In the inflammatory category, Spearman's test showed similar results between raters A, B, and C (rs = 0.47, rs = 0.41, and rs = 0.47, respectively). This study reports the diagnostic validation using digital slides in view of the need to optimize the cytology visualization process. Our experience shows good diagnostic agreement between digital and optical microscopy in several analyzed categories, but mainly in relation to cellular atypia and inflammatory processes.

Horizontal Transposon Transfer and Their Ecological Drivers: The Case of Flower-breeding Drosophila.

Understanding the mechanisms that shape the architecture, diversity, and adaptations of genomes and their ecological and genetic interfaces is of utmost importance to understand biological evolution. Transposable elements (TEs) play an important role in genome evolution, due to their ability to transpose within and between genomes, providing sites of nonallelic recombination. Here we investigate patterns and processes of TE-driven genome evolution associated with niche diversification. Specifically, we compared TE content, TE landscapes, and frequency of horizontal transposon transfers (HTTs) across genomes of flower-breeding Drosophila (FBD) with different levels of specialization on flowers. Further, we investigated whether niche breadth and ecological and geographical overlaps are associated with a potential for HTT rates. Landscape analysis evidenced a general phylogenetic pattern, in which species of the D. bromeliae group presented L-shaped curves, indicating recent transposition bursts, whereas D. lutzii showed a bimodal pattern. The great frequency of highly similar sequences recovered for all FBD suggests that these species probably experienced similar ecological pressures and evolutionary histories that contributed to the diversification of their mobilomes. Likewise, the richness of TEs superfamilies also appears to be associated with ecological traits. Furthermore, the two more widespread species, the specialist D. incompta and the generalist D. lutzii, presented the highest frequency of HTT events. Our analyses also revealed that HTT opportunities are positively influenced by abiotic niche overlap but are not associated with phylogenetic relationships or niche breadth. This suggests the existence of intermediate vectors promoting HTTs between species that do not necessarily present overlapping biotic niches.

Atlantic flower-invertebrate interactions: A data set of occurrence and frequency of floral visits.

Encounters between flowers and invertebrates are key events for the functioning of tropical forests. Assessing the structure of networks composed of the interactions between those partners leads to a better understanding of ecosystem functioning and the effects of environmental factors on ecological processes. Gathering such data is, however, costly and time-consuming, especially in the highly diverse tropics. We aimed to provide a comprehensive repository of available flower-invertebrate interaction information for the Atlantic Forest, a South American tropical forest domain. Data were obtained from published works and "gray literature," such as theses and dissertations, as well as self-reports by co-authors. The data set has ~18,000 interaction records forming 482 networks, each containing between one and 1061 interaction links. Each network was sampled for about 200 h or less, with few exceptions. A total of 641 plant genera within 136 different families and 39 orders were reported, with the most abundant and rich families being Asteraceae, Fabaceae, and Rubiaceae. Invertebrates interacting with these plants were all arthropods from 10 orders, 129 families, and 581 genera, comprising 2419 morphotypes (including 988 named species). Hymenoptera was the most abundant and diverse order, with at least six times more records than the second-ranked order (Lepidoptera). The complete data set shows Hymenoptera interacting with all plant orders and also shows Diptera, Lepidoptera, Coleoptera, and Hemiptera to be important nodes. Among plants, Asterales and Fabales had the highest number of interactions. The best sampled environment was forest (~8000 records), followed by pastures and crops. Savanna, grasslands, and urban environments (among others) were also reported, indicating a wide range of approaches dedicated to collecting flower-invertebrate interaction data in the Atlantic Forest domain. Nevertheless, most reported data were from forest understory or lower strata, indicating a knowledge gap about flower-invertebrate interactions at the canopy. Also, access to remote regions remains a limitation, generating sampling bias across the geographical range of the Atlantic Forest. Future studies in these continuous and hard-to-access forested areas will yield important new information regarding the interactions between flowers and invertebrates in the Atlantic Forest. There are no copyright restrictions on the data set. Please cite this data paper if the data are used in publications and teaching events.

Arundic acid (ONO-2526) inhibits stimulated-S100B secretion in inflammatory conditions.

Astrocytes respond to injury by modifying the expression profile of several proteins, including the S100 calcium-binding protein B (S100B), assumed to be a marker as well as a mediator of brain injury. AA is an inhibitor of S100B synthesis and plays a protective role in different models of brain injury, as decreases in S100B expression cause decreases in extracellular S100B. However, S100B mRNA expression, S100B protein content and S100B secretion do not always occur in association; as such, we herein investigated the effect of AA on S100B secretion, using different approaches with three stimulating conditions for S100B secretion, namely, low potassium medium, TNF-α (in hippocampal slices) and LPS exposure (in astrocyte cultures). Our data indicate that AA directly affects S100B secretion, indicating that the extracellular levels of this astroglial protein may be mediating the action of this compound. More importantly, AA had no effect on basal S100B secretion, but inhibited stimulated S100B secretion (stimulated either by the proinflammatory molecules, LPS or TNF-α, or by low potassium medium). Data from hippocampal slices that were directly exposed to AA, or from animals that received the acid by intracerebroventricular infusion, contribute to understanding its neuroprotective effect.

Evolutionary history and classification of Micropia retroelements in Drosophilidae species.

Transposable elements (TEs) have the main role in shaping the evolution of genomes and host species, contributing to the creation of new genes and promoting rearrangements frequently associated with new regulatory networks. Support for these hypotheses frequently results from studies with model species, and Drosophila provides a great model organism to the study of TEs. Micropia belongs to the Ty3/Gypsy group of long terminal repeats (LTR) retroelements and comprises one of the least studied Drosophila transposable elements. In this study, we assessed the evolutionary history of Micropia within Drosophilidae, while trying to assist in the classification of this TE. At first, we performed searches of Micropia presence in the genome of natural populations from several species. Then, based on searches within online genomic databases, we retrieved Micropia-like sequences from the genomes of distinct Drosophilidae species. We expanded the knowledge of Micropia distribution within Drosophila species. The Micropia retroelements we detected consist of an array of divergent sequences, which we subdivided into 20 subfamilies. Even so, a patchy distribution of Micropia sequences within the Drosophilidae phylogeny could be identified, with incongruences between the species phylogeny and the Micropia phylogeny. Comparing the pairwise synonymous distance (dS) values between Micropia and three host nuclear sequences, we found several cases of unexpectedly high levels of similarity between Micropia sequences in divergent species. All these findings provide a hypothesis to the evolution of Micropia within Drosophilidae, which include several events of vertical and horizontal transposon transmission, associated with ancestral polymorphisms and recurrent Micropia sequences diversification.

First molecular phylogeny of Paralucilia Brauer & Bergenstamm, 1891 (Insecta, Diptera, Calliphoridae): A preliminary approach.

Paralucilia Brauer & Bergenstamm, 1891 (Diptera, Oestroidea, Calliphoridae) is a small genus of blowflies restricted to the Neotropical region, which is commonly reported on decaying corpses and vertebrate carcasses. The number of species currently assigned to this genus and their denominations are contentious, with either three or five species recognized by different authors. This taxonomic instability results in a lack of consensus in species determination, making it impossible to compare results from different studies as well as to elaborate confident taxonomic keys. In order to solve some of the most commonly reported taxonomic conflicts within this genus, to the best of our knowledge, this study presents the first phylogenetic hypothesis for the relationships among Paralucilia species, based on molecular analysis of the COI, ITS2, 28S, and 16S genes. Maximum parsimony, maximum likelihood, and Bayesian inference analyses were used for phylogenetic reconstruction and divergence time estimation analyses. Intra- and interspecific genetic distances were calculated among species using the COI dataset. The results showed that at least three of the five currently accepted species are well defined: P. fulvinota, P. pseudolyrcea, and P. paraensis, however, a significant level of intraspecific variation was observed in P. fulvinota. These findings will assist future revisions of the description, classification, and distribution of species of Paralucilia, as well as in the elaboration of taxonomic keys. Additionally, we show that it is possible to clarify the evolutionary history of this Neotropical genus using supplementary evidence such as morphology and molecular data.

A pilot study on Hla-G locus control region haplotypes and cervical intraepithelial neoplasias.

Human papillomavirus (HPV) can induce cervical intraepithelial neoplasias (CIN) grades 1, 2 and 3. Untreated, these lesions may progress to cervical cancer (CC) which is the third most common cancer in women worldwide. HLA-G plays an immunotolerant role in the immune response. The aim of this study was to characterize the configuration of SNPs located at the distal promoter of HLA-G in patients with CIN2 and CIN3 and control women. The study sample was composed of 207 women as follows: 73 diagnosed with CIN2 lesions, 56 with CIN3 and 78 healthy control women. Genotyping was performed by sequence base typing. Eleven haplotype configurations subdivided in two main haplogroups (H1dist and H2dist), were characterized and compared between patients and controls. The haplotypes H1.1Dist (GAGAACGC) and H2.1Dist (AGGTACAC) were more frequent in Euro-Descendants as well as in Brazilian Mixed. Nevertheless, the haplotype H2.1Dist standed out as a susceptibility haplotype in Brazilian Mixed patients while the H1.1Dist presented a protector effect in this same ethnic group. Whether such LCR haplotype configurations can impact on HLA-G gene expression levels in women who developed cervical intraepithelial neoplasia is still unknown and it is of utmost importance that more investigation on this field be pursued.

The use of DNA barcode for identifying species of Oxysarcodexia Townsend (Diptera: Sarcophagidae): A preliminary survey.

Oxysarcodexia is one of the Neotropical richest genera within the Sarcophagidae family. Medical, veterinary and forensic importance of these flies are due to their association with corpses, cases of myiasis in humans and domestic animals, and being pathogen carriers. Regarding morphological identification, molecular techniques, especially the DNA-based ones, arise as useful alternatives or complementary methodologies for species identification. Thus, in this study we aimed to investigate the potential of the COI marker (barcode region) to delimit Oxysarcodexia species in comparison with the morphological identification criteria. A COI fragment was amplified and the length of the sequences after alignment were of 648bp (149 parsimoniously informative variable sites). According to the Neighbor-Joining phylogenetic tree, specimens of the same morphological species were clustered in monophyletic clades (82-100% bootstrap branch support). Species-level resolution thus achieved was successful, despite low interspecific divergence (1.8-2.3%) and since interspecific variation was higher than intraspecific divergence (0.1-1.2%). Therefore, the use of COI barcode sequences supports differentiation and identification of the Oxysarcodexia species studied.

Schistosomiasis in Southern Brazil 17 years after the confirmation of the first autochthonous case.

INTRODUCTION: Schistosomiasis, a parasitic disease, is an important public health issue in Brazil, particularly Northern Brazil. Since the first recorded occurrence of Biomphalaria glabrata in the States of Santa Catarina and Rio Grande do Sul, it has spread to South Brazil. METHODS: Using the Information System for Notifiable Diseases (SINAN), we assessed the disease spread in Southern Brazil. RESULTS: In Rio Grande do Sul, nine localities had confirmed cases (n = 95, 2001-2013). CONCLUSIONS: We confirmed disease expansion to Southern Brazil. We demonstrated the effectiveness of SINAN to monitor notifiable diseases. Our results are useful to develop preventive actions for schistosomiasis control.

Chromosomal evolution in the Drosophila cardini group (Diptera: Drosophilidae): photomaps and inversion analysis.

Detailed chromosome photomaps are the first step to develop further chromosomal analysis to study the evolution of the genetic architecture in any set of species, considering that chromosomal rearrangements, such as inversions, are common features of genome evolution. In this report, we analyzed inversion polymorphisms in 25 different populations belonging to six neotropical species in the cardini group: Drosophila cardini, D. cardinoides, D. neocardini, D. neomorpha, D. parthenogenetica and D. polymorpha. Furthermore, we present the first reference photomaps for the Neotropical D. cardini and D. parthenogenetica and improved photomaps for D. cardinoides, D. neocardini and D. polymorpha. We found 19 new inversions for these species. An exhaustive pairwise comparison of the polytene chromosomes was conducted for the six species in order to understand evolutionary patterns of their chromosomes.

Efeitos da atividade física na memória declarativa, capacidade funcional e qualidade de vida em idosos / Effects of physical activity in declarative memory, functional capacity and quality of life in elderly

A expectativa de sobrevida da população idosa tem aumentado nos últimos anos, podendo o processo de envelhecimento ser acompanhado por declínio das capacidades funcionais, físicas e cognitivas dos idosos. Dentro da função cognitiva, a memória apresenta-se como uma das suas principais queixas. Os objetivos do estudo foram analisar a memória declarativa, a capacidade funcional e qualidade de vida de idosos ativos e insuficientemente ativos, a fim de observar se a atividade física interfere positivamente nestas variáveis. Trata-se de estudo transversal, do qual fizeram parte dois grupos de comparação, idosos ativos (G1) e insuficientemente ativos (G2), com idade acima de 60 anos, residentes no município de Santa Maria-RS. A avaliação foi composta pelos seguintes instrumentos: Teste de Memória Emocional, Índice de Katz e Avaliação da Qualidade de Vida: SF-36. Como resultados, foi encontrado que os idosos do grupo G1 apresentaram melhores escores tanto na memória como na qualidade de vida quando comparados aos do grupo G2. Já no quesito capacidade funcional para atividades básicas de vida diária, avaliado pelo Índice de Katz, não houve diferença estatisticamente significativa entre os grupos, mas quando observados os resultados da variável capacidade funcional avaliado pelo SF-36, o G1 apresentou melhores resultados com diferença estatisticamente significativa quando comparado ao G2. Devido à importância da atividade física no retardo dos declínios decorrentes do envelhecimento, torna-se necessário criar estratégias para a participação dos idosos em grupos de atividades, contribuindo para a melhoria da qualidade de vida, independência e participação.

Life expectancy of the elderly population has increased in recent years, and the aging process may be accompanied by functional capacity, physical and cognitive decline in the elderly. In cognitive functions, memory appears as one of their main complaints. This study aimed to analyze the declarative memory, functional capacity and quality of life in active and insufficiently active seniors, in order to observe whether physical activity positively affects these variables. This is a cross-sectional study, which included two groups of comparison, active (G1) and insufficiently active seniors (G2), aged over 60, living in the city of Santa Maria-RS. The evaluation consisted of the following instruments: the Emotional Memory Test, the Katz Index and Short Form Health Survey: SF-36. Results showed that the elderly group G1 showed higher scores in both the memory and the quality of life when compared to G2. Concerning the functional capacity for basic activities of daily living, assessed by the Katz Index, there was no statistically significant difference between groups, but when we observe the results of the functional capacity variable assessed by the SF-36, G1 showed better results with statistically significant differences as compared with G2. Because of the importance of physical activity in the delay of the declines from aging, it is necessary to create strategies for the participation of the elderly in group activities, contributing to the improvement of quality of life, independence and participation.

Experience of family members as a result of children's hospitalization at the Intensive Care Unit / Experiencia vivida por los familiares con la internación de niños en la Unidad de Cuidado Intensivo / Experiência vivida pelos familiares com a internação de crianças na unidade de terapia intensiva

Objective. To describe the experience of family members as a result of children's hospitalization at the Intensive Care Unit (ICU). Methodology. Descriptive and cross-sectional study. A structured interview was held with 20 relatives of patients hospitalized at two clinics of the Botucatu Medical School at Universidade Estadual Paulista 'Júlio de Mesquita Filho'. Information was collected between July and September 2010. Results. The main characteristics of the participating relatives were: 80% mothers of the children; 70% low education level and 70% married. Sixty percent of the children were hospitalized at the ICU for the first time. Eighty percent of the interviewees believe that the children's behavior changes inside the unit and 85% consider that visiting hours are sufficient. The predominant negative feelings are fear (50%) and insecurity (20%), while the predominant positive feelings are hope (50%) and the expectation of discharge (25%). The professional who most supported the relatives was the nurse (35%). Conclusion. The family members' experience as a result of the children's hospitalization at the ICU involves positive and negative aspects, which also affect the child's behavior at the unit.

Objetivo. Describir la experiencia vivida por los familiares con la internación de niños en la Unidad de Cuidados Intensivos (UCI). Metodología. Estudio descriptivo de tipo transversal. Se realizó una entrevista estructurada a 20 familiares de pacientes hospitalizados en dos clínicas de la Facultad de Medicina de Botucatu de la Universidade Estadual Paulista 'Júlio de Mesquita Filho'. La información se recolectó entre julio y septiembre de 2010. Resultados. Las principales características de los familiares participantes fueron: 80%, madres de los niños; 70% tenía escolaridad baja y el 70% estaba casado. Fue la primera hospitalización en la UCI en el 60% de los niños. El 80% de los entrevistados cree que hay cambio en el comportamiento del niño dentro de la unidad y el 85% considera suficiente el tiempo de visita. Los sentimientos negativos predominantes son el miedo (50%) y la inseguridad (20%), mientras que los positivos son la esperanza (50%) y la expectativa del alta (25%). El profesional que más apoyo brindó al familiar fue el enfermero (35%). Conclusión. La experiencia de los familiares con la internación del niño en UCI tiene aspectos positivos y negativos, que incluso, afectan el comportamiento del niño en la unidad.

Objetivo. Descrever a experiência vivida pelos familiares com a internação de crianças na Unidade de Terapia Intensiva (UTI). Metodologia. Estudo descritivo de tipo transversal. Realizou-se uma entrevista estruturada a 20 familiares de pacientes hospitalizados em duas clínicas da Faculdade de Medicina de Botucatu da Universidade Estadual Paulista 'Júlio de Mesquita Filho'. A informação se coletou entre julho e setembro de 2010. Resultados. As principais características dos familiares participantes foram: 80% eram mães das crianças, 70% tinham escolaridade baixa e o 70% estavam casados. Foi a primeira hospitalização na UTI em 60% das crianças. 80% dos entrevistados creem que há mudança no comportamento da criança dentro da unidade e 85% considera suficiente o tempo de visita. Os sentimentos negativos predominantes são o medo (50%) e a insegurança (20%), enquanto os positivos são a esperança (50%) e a expectativa do alta (25%). O profissional que mais apoio brindou ao familiar foi o enfermeiro (35%). Conclusão. A experiência dos familiares com a internação da criança na UTI tem aspectos positivos e negativos, que inclusive, afetam o comportamento da criança na unidade.

The LTR retrotransposon micropia in the cardini group of Drosophila (Diptera: Drosophilidae): a possible case of horizontal transfer.

The presence of the micropia retroelement from the Ty1-copia family of LTR retroelements was investigated in three species of the Drosophila cardini group. Southern blot analysis suggested the existence of at least four micropia copies in the genomes of D. cardinoides, D. neocardini and D. polymorpha populations. The high sequence similarity between dhMiF2 and Dm11 clones (micropia retroelements isolated from D. hydei and D. melanogaster, respectively) with micropia sequences amplified from D. cardini group genome supports the hypothesis that this retroelement plays an active role in horizontal transfer events between D. hydei and the D. cardini group.

Study of the Drosophilidae (Diptera) communities on Atlantic Forest islands of Santa Catarina State, Brazil.

A study of the community dynamics of Drosophilidae was carried out in six insular communities and two others on the mainland. Seasonal collections were carried out throughout two years in Santa Catarina State, southern of Brazil. The diversity index calculations show high values when compared with temperate climate communities. The sites on the mainland (Serra do Tabuleiro) presented the highest diversity, which was measured by the Diversity Index (H'). These sites are covered by primary Atlantic Forest and theoretically should have a higher variation of ecological niches. A dendogram showing the similarity between the communities, calculated by Morisita Index, points to a level of similarity equal to 60% for all communities. In this diagram, we can see two clades: one on the mainland and the other on the islands. The six island sites are grouped into one clade and separated into two subclades, one including the sites on Santa Catarina Island and the other consisting of the islands adjacent to this last and very much larger one. These groupings show the very important role of the spatial component on the prediction of the structure of the communities. This fact raises the discussion about the high complexity of the Atlantic Forest ecosystem and consequently the unpredictability of its fauna, highlighting the need of its conservation.