Detection of arboviruses in Aedes aegypti through transovarian analysis: A study in Goiânia, Goiás.

BACKGROUND: Arboviral diseases are a group of infectious diseases caused by viruses transmitted by arthropods, mainly mosquitoes. These diseases, such as those caused by the dengue (DENV), Zika (ZIKV), chikungunya (CHIKV), and yellow fever (YFV) viruses, have a significant impact worldwide. In this context, entomological surveillance plays a crucial role in the control and prevention of arboviruses by providing essential information on the presence, distribution, and activity of vector mosquitoes. Based on entomological surveillance, transovarian transmission provides information regarding the maintenance and dissemination of arboviruses. The objective of this study was to detect these arboviruses in Goiânia, Goiás, and analyze the occurrence of transovarian transmission. METHODS: Aedes aegypti eggs were collected from different regions of Goiânia and cultivated under controlled laboratory conditions until the emergence of adult mosquitoes. Adult females were grouped into pools containing their heads and thoraxes. These pools were subsequently evaluated using reverse-transcription quantitative polymerase chain reaction (RT-qPCR) assay. RESULTS: A total of 157 pools (N=1570) were analyzed, with two pools testing positive for CHIKV and one pool testing positive for ZIKV, indicating that the offspring resulting from transovarian transmission are potentially infectious. CONCLUSIONS: In summary, the demonstration of the vertical transmission mechanisms of CHIKV and ZIKV in A. aegypti serves as an alert to health authorities, as these diseases are still underreported, and their primary urban vector has likely acquired this capacity, contributing to the dissemination of these infections.

ROR1 expression in mature B lymphoid neoplasms by flow cytometry.

Immunophenotyping by flow cytometry is an integral part of the diagnosis and classification of leukemias/lymphomas. The expression of ROR1 associated with chronic B lymphocytic leukemia (CLL) is well described in the literature, both in its diagnosis and in the follow-up of minimal residual disease (MRD) research, however, there are few studies regarding the expression pattern of ROR1 in other subtypes of mature B lymphoid neoplasms. With the aim of evaluating the expression of ROR1 and associating it with the expression of other important markers for the differentiation of mature B lymphoid neoplasms (MBLN), 767 samples of cases that entered our laboratory for immunophenotyping with clinical suspicion of MBLN were studied. ROR1 expression is predominant in CD5+/CD10- neoplasms. Overall, positive ROR1 expression was observed in 461 (60.1%) cases. The CD5+/CD10- group had a significantly higher proportion of ROR1 positive samples (89.9%) and more brightly expressed ROR1 than the other groups. Our results highlight the importance of evaluating ROR1 expression in the diagnosis of MBLN to contribute to the differential diagnosis, and possibly therapy of mainly CLL, and indicate that this marker could be considered as a useful addition to immunophenotypic panels, particularly for more challenging cases.

Detection of arboviruses in Aedes aegypti through transovarian analysis: A study in Goiânia, Goiás

Abstract Background: Arboviral diseases are a group of infectious diseases caused by viruses transmitted by arthropods, mainly mosquitoes. These diseases, such as those caused by the dengue (DENV), Zika (ZIKV), chikungunya (CHIKV), and yellow fever (YFV) viruses, have a significant impact worldwide. In this context, entomological surveillance plays a crucial role in the control and prevention of arboviruses by providing essential information on the presence, distribution, and activity of vector mosquitoes. Based on entomological surveillance, transovarian transmission provides information regarding the maintenance and dissemination of arboviruses. The objective of this study was to detect these arboviruses in Goiânia, Goiás, and analyze the occurrence of transovarian transmission. Methods: Aedes aegypti eggs were collected from different regions of Goiânia and cultivated under controlled laboratory conditions until the emergence of adult mosquitoes. Adult females were grouped into pools containing their heads and thoraxes. These pools were subsequently evaluated using reverse-transcription quantitative polymerase chain reaction (RT-qPCR) assay. Results: A total of 157 pools (N=1570) were analyzed, with two pools testing positive for CHIKV and one pool testing positive for ZIKV, indicating that the offspring resulting from transovarian transmission are potentially infectious. Conclusions: In summary, the demonstration of the vertical transmission mechanisms of CHIKV and ZIKV in A. aegypti serves as an alert to health authorities, as these diseases are still underreported, and their primary urban vector has likely acquired this capacity, contributing to the dissemination of these infections.

Immunostimulatory nanoparticles delivering cytokines as a novel cancer nanoadjuvant to empower glioblastoma immunotherapy.

Glioblastoma (GBM) stands as a highly aggressive and deadly malignant primary brain tumor with a median survival time of under 15 months upon disease diagnosis. While immunotherapies have shown promising results in solid cancers, brain cancers are still unresponsive to immunotherapy due to immunological dysfunction and the presence of a blood-brain barrier. Interleukin-12 (IL-12) emerges as a potent cytokine in fostering anti-tumor immunity by triggering interferon-gamma production in T and natural killer cells and changing macrophages to a tumoricidal phenotype. However, systemic administration of IL-12 toxicity in clinical trials often leads to significant toxicity, posing a critical hurdle. To overcome this major drawback, we have formulated a novel nanoadjuvant composed of immunostimulatory nanoparticles (ISN) loaded with IL-12 to decrease IL-12 toxicity and enhance the immune response by macrophages and GBM cancer cells. Our in vitro results reveal that ISN substantially increase the production of pro-inflammatory cytokines in GBM cancer cells (e.g. 2.6 × increase in IL-8 expression compared to free IL-12) and macrophages (e.g. 2 × increase in TNF-α expression and 6 × increase in IL-6 expression compared to the free IL-12). These findings suggest a potential modulation of the tumor microenvironment. Additionally, our study demonstrates the effective intracellular delivery of IL-12 by ISN, triggering alterations in the levels of pro-inflammatory cytokines at both transcriptional and protein expression levels. These results highlight the promise of the nanoadjuvant as a prospective platform for resharing the GBM microenvironment and empowering immunotherapy.

Report of natural Mayaro virus infection in Mansonia humeralis (Dyar & Knab, Diptera: Culicidae).

BACKGROUND: Arboviruses are a group of viruses transmitted to vertebrate hosts by certain blood-feeding arthropods. Among urban vectors of arboviruses, mosquitoes of the genus Aedes are the most common. However, other mosquitoes may be susceptible to infection and involved in the transmission, such as Mansonia spp. Therefore, this study aimed to investigate whether Mansonia humeralis can be infected with the Mayaro virus (MAYV). METHODS: These insects were collected from 2018 to 2020 in chicken coops of rural communities in Jaci Paraná in Porto Velho, Rondônia, Brazil, while performing blood-feeding on roosters. The mosquitoes were randomly grouped in pools from which the head and thorax were macerated and checked for the presence of MAYV by quantitative reverse transcription polymerase chain reaction (RT-qPCR). The positive pools were used to infect the C6/36 cell line, and on different days post-infection, the supernatant of the infected cells was subjected to viral detection by RT-qPCR. RESULTS: A total of 183 pools of female mosquitoes were tested, of which 18% were positive for MAYV; some samples from insect pools inoculated into C6/36 cells showed in vitro multiplication capacity between 3 and 7 days post-infection. CONCLUSIONS: This is the first report of Ma. humeralis mosquitoes that are naturally infected by MAYV, indicating that these vectors may be potential transmitting agents of this arbovirus.

Vocational Rehabilitation and Length of Stay at Work after Work-Related Musculoskeletal Disorders: A Longitudinal Study in Brazil.

Vocational rehabilitation is an intervention to enhance the return to work and improve quality of life. The aim of this study was to evaluate sociodemographic and occupational factors associated with the length of stay at work among workers with work-related musculoskeletal disorders (WRMDs) who had undergone rehabilitation through the Brazilian public social security system. This was a longitudinal study among 680 workers with histories of disability due to WRMDs who returned to the formal job market after vocational rehabilitation between 2014 and 2018. Survival analysis was performed to identify the factors influencing permanence in work. Job dismissal occurred for 29.26% of the workers. The average duration of employment after returning to the formal job position was 56 months. The following factors were associated with shorter length of employment: living in the southeastern region (HR: 2.78; 95% CI 1.12-6.91) or southern region (HR: 2.68; 95% CI 1.04-6.90) of Brazil; working in transportation, storage or postal services (HR: 2.57; 95% CI 1.07-6.17); or working in financial activities, insurance or related services (HR: 2.70; 95% CI 1.05-6.89). These findings may contribute to the discussion about prevention of disability and interventions to ensure health care for workers with WRMD disabilities who undergo rehabilitation.

Procedures for health data linkage: applications in health surveillance. / Procedimentos para vinculação de dados da saúde: aplicações na vigilância em saúde.

OBJECTIVE: To present a standardized methodology for linking different public health databases. METHODS: This was a methodological review article specifically describing data processing procedures for deterministic linkage between structured databases. It instructs on how to: treat data, select linkage keys, and link databases using two databases simulated in R software. RESULTS: The commands used for the deterministic linkage of the inner_join type were presented. The linkage process resulted in a database with 40,108 pairs using only the "Name" key. Adding the second key, "Name of mother", the resulted dropped to 112 pairs. By adding the third key, "Date of birth", only two pairs were identified. CONCLUSION: Database linkage and its analysis are valid and valuable tools for health services in supporting health surveillance actions.

Polymeric nanoparticles targeting Sialyl-Tn in gastric cancer: A live tracking under flow conditions.

Drug delivery using nanoparticles (NPs) represents a potential approach for therapy in cancer, such gastric cancer (GC) due to their targeting ability and controlled release properties. The use of advanced nanosystems that deliver anti-cancer drugs specifically to tumor cells may strongly rely on the expression of cancer-associated targets. Glycans aberrantly expressed by cancer cells are attractive targets for such delivery strategy. Sialylated glycans, such as Sialyl-Tn (STn) are aberrantly expressed in several epithelial tumors, including GC, being a potential target for a delivery nanosystem. The aim of this study was the development of NPs surface-functionalized with a specific antibody targeting the STn glycan and further evaluate this nanosystem effectiveness regarding its specificity and recognition capacity. Our results showed that the NPs surface-functionalized with anti-STn antibody efficiently are recognized by cells displaying the cancer-associated STn antigen under static and live cell monitoring flow conditions. This uncovers the potential use of such NPs for drug delivery in cancer. However, flow exposure was disclosed as an important biomechanical parameter to be taken into consideration. Here we presented an innovative and successful methodology to live track the NPs targeting STn antigen under shear stress, simulating the physiological flow. We demonstrate that unspecific binding of NPs agglomerates did not occur under flow conditions, in contrast with static assays. This robust approach can be applied for in vitro drug studies, giving valuable insights for in vivo studies.

Brain-Targeted Drug Delivery.

At present, brain diseases affect one in six people worldwide, and they include a wide range of neurological diseases from Alzheimer's and Parkinson's diseases to epilepsy, brain injuries, brain cancer, neuroinfections and strokes [...].

Torulaspora delbrueckii Phenotypic and Metabolic Profiling towards Its Biotechnological Exploitation.

Wine is a particularly complex beverage resulting from the combination of several factors, with yeasts being highlighted due to their fundamental role in its development. For many years, non-Saccharomyces yeasts were believed to be sources of spoilage and contamination, but this idea was challenged, and many of these yeasts are starting to be explored for their beneficial input to wine character. Among this group, Torulaspora delbrueckii is gaining relevance within the wine industry, owing to its low volatile acidity production, increased release of aromatic compounds and enhanced color intensity. In addition, this yeast was also attracting interest in other biotechnological areas, such as bread and beer fermentation. In this work, a set of 40 T. delbrueckii strains, of varied geographical and technological origins, was gathered in order to characterize the phenotypic behavior of this species, focusing on different parameters of biotechnological interest. The fermentative performance of the strains was also evaluated through individual fermentations in synthetic grape must with the isolates' metabolic profile being assessed by HPLC. Data analysis revealed that T. delbrueckii growth is significantly affected by high temperature (37 °C) and ethanol concentrations (up to 18%), alongside 1.5 mM SO2, showing variable fermentative power and yields. Our computation models suggest that the technological origin of the strains seems to prevail over the geographical origin as regards the influence on yeast properties. The inter-strain variability and profile of the products through the fermentative processes reinforce the potential of T. delbrueckii from a biotechnological point of view.

Structural and functional analysis of broad pH and thermal stable protease from Penicillium aurantiogriseum URM 4622.

This study aimed to better characterize a recently purified stable extracellular alkaline peptidase produced by Penicillium aurantiogriseum (URM 4622) through fluorescence spectroscopy, far-UV circular dichroism, kinetic and thermodynamic models to understand its' structure-activity and denaturation. Fluorescence data showed that changing pH leads to tryptophan residues exposure to more hydrophilic environments at optimum activity pH 9.0 and 10.0. When thermally treated, it displayed less unfolding at these pH values, along with 4-fold less photoproducts formation than at neutral pH. Different pH CD spectra showed more ß-sheet (21.5-43.0%) than α-helix (1-6.2%). At pH9.0, more than 2-fold higher α-helix content than any other pH. The melting temperature (Tm) was observed between 50 and 60 °C at all pH studied, with lower Tm at pH 9.0-11.0 (54.9-50.3 °C). The protease displayed two phase transition, with two energies of denaturation, and a 4-fold higher thermal stability (ΔH°m) than reports for other microorganism's proteases. An irreversible folding transition occurs between 50 and 60 °C. It displayed energies of denaturation suggesting higher thermal stability than reported for other microorganism's proteases. These results help elucidating the applicability of this new stable protease.

Procedimentos para vinculação de dados da saúde: aplicações na vigilância em saúde / Procedimientos para la vinculación de datos sanitarios: aplicaciones en la vigilancia en la salud / Procedures for health data linkage: applications in health surveillance

Objetivo: Apresentar metodologia padronizada para vinculação de diferentes bancos de dados em saúde pública. Métodos: Artigo de revisão metodológica, com descrição específica de processos de tratamento de dados para vinculação (linkage) determinística entre bancos de dados estruturados. Instruiu-se como tratar os dados, selecionar chaves de vinculação e vincular os bancos, utilizando-se dois bancos de dados simulados no software R. Resultados: Foram apresentados os comandos utilizados para a vinculação determinística, do tipo inner_join. O processo de vinculação resultou em um banco de dados com 40.108 pares ao se utilizar apenas a chave "Nome". Com a adição da segunda chave, "Nome da mãe", o resultado caiu para 112 pares. Ao adicionar a terceira chave, "Data de nascimento", apenas dois pares foram identificados. Conclusão: A vinculação de bancos de dados e suas análises são ferramentas válidas e úteis para os serviços de saúde, no apoio a ações de vigilância em saúde.

Objetivo: Presentar metodología estandarizada para vincular diferentes bases de datos de salud pública. Métodos: Artículo de revisión metodológica y descripción de los procesos de tratamiento de datos para la vinculación determinista entre bases de datos. Se dieron instrucciones sobre como manejar los datos, seleccionar claves de vinculación y vincular las bases de datos empleando dos bases de datos simuladas en el software R. Resultados: Se presentaron los comandos utilizados para la vinculación determinista, del tipo inner-join. El proceso resultó en una base de datos con 40.108 pares utilizando únicamente la clave "Nombre". Con la adición de la segunda clave, "Nombre de la madre", el resultado se redujo a 112 pares. Al agregar la tercera clave, "Fecha de nacimiento", solo se identificaron dos pares. Conclusión: La vinculación de bases de datos y sus análisis son herramientas válidas y útiles para que los servicios de salud las utilicen para apoyar las acciones de vigilancia en la salud.

Objective: To present a standardized methodology for linking different public health databases. Methods: This was a methodological review article specifically describing data processing procedures for deterministic linkage between structured databases. It instructs on how to: treat data, select linkage keys, and link databases using two databases simulated in R software. Results: The commands used for the deterministic linkage of the inner_join type were presented. The linkage process resulted in a database with 40,108 pairs using only the "Name" key. Adding the second key, "Name of mother", the resulted dropped to 112 pairs. By adding the third key, "Date of birth", only two pairs were identified. Conclusion: Database linkage and its analysis are valid and valuable tools for health services in supporting health surveillance actions.

Conhecimento da equipe multiprofissional sobre segurança do paciente em Unidade de Terapia Intensiva / Knowledge of the multidisciplinary team on patient safety in an Intensive Care Unit

Analisar os fatores que interferem na segurança do paciente em Unidades de Terapia Intensiva (UTI) mediante o conhecimento da equipe multiprofissional. Tratou-se de um estudo descritivo com abordagem quantiqualitativa realizado com 40 profissionais de saúde, que compõe a equipe multiprofissional da UTI de um hospital estadual, referência regional. A coleta de dados foi realizada entre os meses de maio a dezembro de 2020. Deu-se por meio de entrevista semiestruturada, direcionada por um roteiro temático e questionário sociodemográfico. Os dados foram processados no software IRAMUTEQ, através da Classificação Hierárquica Descendente e analisados por meio da técnica do discurso do sujeito coletivo. Foi evidenciado que a fragilidade no conhecimento sobre o conceito de segurança do paciente, a falta de protocolos assistências, os problemas interpessoais, a ocorrência de eventos adversos, a subnotificação e dimensionamento inadequado são alguns dos fatores que interferem na segurança do paciente na UTI. Nota-se a necessidade de resolução das falhas existentes, sobretudo ao que refere ao dimensionamento de pessoal, as questões relacionadas à interação da equipe multiprofissional e a implementação de protocolos institucionais, que são norteadores do cuidado. Com o fortalecimento da cultura de segurança, onde os profissionais sintam-se empoderados a realizar a notificação, não apenas dos EA, como também das circunstâncias notificáveis.

The aim of this study was to analyze the factors that interfere with patient safety in Intensive Care Units (ICU) through the knowledge of the multidisciplinary team. This was a descriptive study with a quantitative and qualitative approach, carried out with 40 health professionals, who make up the multidisciplinary team of the ICU of a regional reference hospital of the state. Data collection was carried out between the months of May and December 2020. It took place through a semi-structured interview, guided by a thematic script and a sociodemographic questionnaire. Data were processed using IRAMUTEQ software, through Descending Hierarchical Classification, and were analyzed using the collective subject discourse technique. It was demonstrated that the lack of knowledge about the concept of patient safety, the lack of care protocols, interpersonal problems, the occurrence of adverse events, underreporting, and inadequate estimations are some of the factors that interfere with patient safety in the ICU. There is a need to resolve existing flaws, especially regarding personnel size, issues related to the interaction of the multidisciplinary team, and the implementation of institutional protocols, which guide care. By strengthening the safety culture, professionals feel empowered to report not only AEs, but also reportable circumstances.

Biotechnological Importance of Torulaspora delbrueckii: From the Obscurity to the Spotlight.

Torulaspora delbrueckii has attracted interest in recent years, especially due to its biotechnological potential, arising from its flavor- and aroma-enhancing properties when used in wine, beer or bread dough fermentation, as well as from its remarkable resistance to osmotic and freezing stresses. In the present review, genomic, biochemical, and phenotypic features of T. delbrueckii are described, comparing them with other species, particularly with the biotechnologically well-established yeast, Saccharomyces cerevisiae. We conclude about the aspects that make this yeast a promising biotechnological model to be exploited in a wide range of industries, particularly in wine and bakery. A phylogenetic analysis was also performed, using the core proteome of T. delbrueckii, to compare the number of homologous proteins relative to the most closely related species, understanding the phylogenetic placement of this species with robust support. Lastly, the genetic tools available for T. delbrueckii improvement are discussed, focusing on adaptive laboratorial evolution and its potential.

Intratumoral VEGF nanotrapper reduces gliobastoma vascularization and tumor cell mass.

Glioblastoma multiforme (GBM) is the most aggressive and invasive malignant brain cancer. GBM is characterized by a dramatic metabolic imbalance leading to increased secretion of the pro-angiogenic factor VEGF and subsequent abnormal tumor vascularization. In 2009, FDA approved the intravenous administration of bevacizumab, an anti-VEGF monoclonal antibody, as a therapeutic agent for patients with GBM. However, the number of systemic side effects and reduced accessibility of bevacizumab to the central nervous system and consequently to the GBM tumor mass limited its effectiveness in improving patient survival. In this study, we combined experimental and computational modelling to quantitatively characterize the dynamics of VEGF secretion and turnover in GBM and in normal brain cells and simultaneous monitoring of vessel growth. We showed that sequestration of VEGF inside GBM cells, can be used as a novel target for improved bevacizumab-based therapy. We have engineered the VEGF nanotrapper, a cargo system that allows cellular uptake of bevacizumab and inhibits VEGF secretion required for angiogenesis activation and development. Here, we show the therapeutic efficacy of this nanocargo in reducing vascularization and tumor cell mass of GBM in vitro and in vivo cancer models.

Host-specific gene expression as a tool for introduction success in Naupactus parthenogenetic weevils.

Food resource access can mediate establishment success in invasive species, and generalist herbivorous insects are thought to rely on mechanisms of transcriptional plasticity to respond to dietary variation. While asexually reproducing invasives typically have low genetic variation, the twofold reproductive capacity of asexual organisms is a marked advantage for colonization. We studied host-related transcriptional acclimation in parthenogenetic, invasive, and polyphagous weevils: Naupactus cervinus and N. leucoloma. We analyzed patterns of gene expression in three gene categories that can mediate weevil-host plant interactions through identification of suitable host plants, short-term acclimation to host plant defenses, and long-term adaptation to host plant defenses and their pathogens. This approach employed comparative transcriptomic methods to investigate differentially expressed host detection, detoxification, immune defense genes, and pathway-level gene set enrichment. Our results show that weevil gene expression responses can be host plant-specific, and that elements of that response can be maintained in the offspring. Some host plant groups, such as legumes, appear to be more taxing as they elicit a complex gene expression response which is both strong in intensity and specific in identity. However, the weevil response to taxing host plants shares many differentially expressed genes with other stressful situations, such as host plant cultivation conditions and transition to novel host, suggesting that there is an evolutionarily favorable shared gene expression regime for responding to different types of stressful situations. Modulating gene expression in the absence of other avenues for phenotypic adaptation may be an important mechanism of successful colonization for these introduced insects.

In vitro model for predicting the access and distribution of drugs in the brain using hCMEC/D3 cells.

The BBB is a protective entity that prevents external substances from reaching the CNS but it also hinders the delivery of drugs into the brain when they are needed. The main objective of this work was to improve a previously proposed in vitro cell-based model by using a more physiological cell line (hCMEC/D3) to predict the main pharmacokinetic parameters that describe the access and distribution of drugs in the CNS: Kpuu,brain, fu,plasma, fu,brain and Vu,brain. The hCMEC/D3 permeability of seven drugs was studied in transwell systems under different conditions (standard, modified with albumin and modified with brain homogenate). From the permeability coefficients of those experiments, the parameters mentioned above were calculated and four linear IVIVCs were established. The best ones were those that relate the in vitro and in vivo Vu,brain and fu,brain (r2 = 0.961 and r2 = 0.940) which represent the binding rate of a substance to the brain tissue, evidencing the importance of using brain homogenate to mimic brain tissue when an in vitro brain permeability assay is done. This methodology could be a high-throughput screening tool in drug development to select the CNS promising drugs in three different in vitro BBB models (hCMEC/D3, MDCK and MDCK-MDR1).

How I investigate minimal residual disease in acute lymphoblastic leukemia.

Minimal Residual Disease (MRD) is the most important independent prognostic factor in acute lymphoblastic leukemia (ALL) and refers to the deep level of measurable disease in cases with complete remission by conventional pathologic analysis, especially by cytomorphology. MRD can be detected by multiparametric flow cytometry, molecular approaches such as quantitative polymerase chain reaction for immunoglobulin and T-cell receptor (IG/TR) gene rearrangements or fusion genes transcript, and high-throughput sequencing for IG/TR. Despite the proven clinical usefulness in detecting MRD, these methods have differences in sensitivity, specificity, applicability, turnaround time and cost. Knowing and understanding these differences, as well as the principles and limitations of each technology, is essential to laboratory standardization and correct interpretation of MRD results in line with treatment time points, therapeutic settings, and clinical trials. Here, we review the methodological approaches to measure MRD in ALL and discuss the advantages and limitations of the most commonly used techniques.