The KMT2A recombinome of acute leukemias in 2023.

Chromosomal rearrangements of the human KMT2A/MLL gene are associated with de novo as well as therapy-induced infant, pediatric, and adult acute leukemias. Here, we present the data obtained from 3401 acute leukemia patients that have been analyzed between 2003 and 2022. Genomic breakpoints within the KMT2A gene and the involved translocation partner genes (TPGs) and KMT2A-partial tandem duplications (PTDs) were determined. Including the published data from the literature, a total of 107 in-frame KMT2A gene fusions have been identified so far. Further 16 rearrangements were out-of-frame fusions, 18 patients had no partner gene fused to 5'-KMT2A, two patients had a 5'-KMT2A deletion, and one ETV6::RUNX1 patient had an KMT2A insertion at the breakpoint. The seven most frequent TPGs and PTDs account for more than 90% of all recombinations of the KMT2A, 37 occur recurrently and 63 were identified so far only once. This study provides a comprehensive analysis of the KMT2A recombinome in acute leukemia patients. Besides the scientific gain of information, genomic breakpoint sequences of these patients were used to monitor minimal residual disease (MRD). Thus, this work may be directly translated from the bench to the bedside of patients and meet the clinical needs to improve patient survival.

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro / Efeitos das redes extracelulares dos neutrófilos durante a infecção in vitro pelo vírus sincicial respiratório

The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.5–16 μg/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,5–16 μg/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios [...].

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro

Abstract The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.516 g/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

Resumo O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,516 g/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios semelhantes àqueles induzidos pelo vírus em células não tratadas, indicando danos citotóxicos e deletérios. Nossos resultados corroboram achados relatados anteriormente de que as NETs contribuem para a imunopatologia desenvolvida por pacientes infectados com hRSV.

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro / Efeitos das redes extracelulares dos neutrófilos durante a infecção in vitro pelo vírus sincicial respiratório

Abstract The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.5-16 μg/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

Resumo O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,5-16 μg/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios semelhantes àqueles induzidos pelo vírus em células não tratadas, indicando danos citotóxicos e deletérios. Nossos resultados corroboram achados relatados anteriormente de que as NETs contribuem para a imunopatologia desenvolvida por pacientes infectados com hRSV.

Sacroiliac joint dysfunction and lumbar pain. Evaluation in a Brazilian population.

The study intended to evaluate the incidence and evolution of sacroiliac joint dysfunction (SIJD). To reach 50 patients with SIJD diagnosis, 192 patients with low back pain and failure in conservative approach were consecutively examined (26% incidence). Initially patients underwent intra-articular (IA) corticosteroid sacroiliac joint (SIJ) block followed, if necessary, by cooled SIJ radiofrequency or referred to surgical intervention, in order of complexity. From the 50 patients submitted to IA SI block, 41 (82%) referred pain and quality of life improvement and lesser rescue analgesics consumption for 25 weeks. The block induced a prompt onset of pain relief and there was a drop in mean pain score from 8 to 2 cm (p < 0.001) maintained up to 25 weeks. Rescue analgesic consumption also significantly dropped (p < 0.05). However, nine patients (18%) did not refer long lasting improvement in the third week evaluation and underwent cooled radiofrequency. From this population of nine, seven were successful (78%) while two were recommended surgery. In view of the 50 patients, 82% were comfortable after IA block, 18% were submitted to radiofrequency, with a success rate of 78%. The final incidence of surgery suggestion was 4%.

El estudio pretende evaluar la incidencia y evolución de la disfunción de la articulación sacroilíaca (DASI). Para llegar a 50 pacientes con diagnóstico de DASI, se examinaron consecutivamente 192 pacientes con dolor lumbar y fracaso en el abordaje conservador (26% de incidencia). Inicialmente, los pacientes se sometieron a un bloqueo de la articulación sacroilíaca (ASI) con corticosteroides intraarticulares (IA) seguido, si era necesario, de radiofrecuencia ASI enfriada o remitidos a una intervención quirúrgica, en orden de complejidad. De los 50 pacientes sometidos al bloqueo IA SI, 41 (82%) refirieron mejoría del dolor y de la calidad de vida y menor consumo de analgésicos de rescate durante 25 semanas. El bloqueo indujo un rápido inicio del alivio del dolor y hubo una caída en la puntuación media del dolor de 8 a 2 cm (p < 0.001) mantenida hasta 25 semanas. El consumo de analgésicos de rescate también disminuyó significativamente (p < 0.05). Sin embargo, nueve pacientes (18%) no refirieron una mejoría duradera en la evaluación de la tercera semana y se sometieron a radiofrecuencia fría. De esta población de nueve, siete tuvieron éxito (78%), mientras que a dos se les recomendó cirugía. De los 50 pacientes, 82% se sintió cómodo después del bloqueo IA, 18% fue sometido a radiofrecuencia, con una tasa de éxito de 78%. La incidencia final de sugerencia de cirugía fue de 4%.

Changes in gut-brain axis parameters in adult rats of both sexes with different feeding pattern that were early nicotine-exposed.

Nicotine is an endocrine disruptor and imprinting factor during breastfeeding that can cause food intake imbalance in the adulthood. As nicotine affects the intestinal microbiota, altering the composition of the bacterial communities and short-chain fatty acids (SCFAs) synthesis in a sex-dependent manner, we hypothesized that nicotine could program the gut-brain axis, consequently modifying the eating pattern of adult male and female rats in a model of maternal nicotine exposure (MNE) during breastfeeding. Lactating Wistar rat dams received minipumps that release 6 mg/kg/day of nicotine (MNE group) or saline for 14 days. The progeny received standard diet from weaning until euthanasia (26 weeks of age). We measured: in vivo electrical activity of the vagus nerve; c-Fos expression in the nucleus tractus solitarius, gastrointestinal peptides receptors, intestinal brain-derived neurotrophic factor (BDNF), SCFAs and microbiota. MNE females showed hyperphagia despite normal adiposity, while MNE males had unchanged food intake, despite obesity. Adult MNE offspring showed decreased Bacteroidetes and increased Firmicutes, Actinobacteria and Proteobacteria. MNE females had lower fecal acetate while MNE males showed higher vagus nerve activity. In summary nicotine exposure through the milk induces long-term intestinal dysbiosis, which may affect eating patterns of adult offspring in a sex-dependent manner.

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro.

The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.5-16 µg/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

Moringa Oleifera: an alternative ingredient to improve the egg quality of Japanese quail / [Moringa oleifera: um ingrediente alternativo na melhora da qualidade de ovos de codornas japonesas]

This study aims to evaluate effects of inclusion of moringa in Japanese quail diets on laying performance, egg quality, blood parameters, serum biochemical profile, and behavior. One hundred and forty-four Japanese quails, approximately 35-d-old, were distributed in a completely randomized design with four treatments and six replications. Treatments were diet inclusion of 0, 2, 4 and 6% of dried and ground moringa leaves. The inclusion of moringa in diets reduces the quadratic feed intake up to the level of 1.20%, increases weight of eggs with a quadratic behavior up to 3.80%, and linearly increases yolk weight. Yolk color changes with higher levels of inclusion of moringa and resulted in more intense colors. The biochemical profile of quails changed slightly but remains within the normal range. The inclusion of 4% of moringa in diets increases alkaline phosphatase. Regarding cholesterol and triglycerides, diet with 6% moringa inclusion was lower when compared to the others. The behavior of laying quails does not change due to inclusion of moringa in diets. Up to 3.83% of Moringa oleifera can be included in Japanese quail diet to improve egg quality without compromising performance, biochemical profile, blood parameters and behavior.(AU)

O objetivo deste estudo foi avaliar a inclusão de Moringa oleifera na dieta de codornas japonesas sobre o desempenho, a qualidade de ovos, os parâmetros sanguíneos, o perfil bioquímico e o comportamento. Foram utilizadas 144 codornas japonesas, com aproximadamente 35 dias de idade, as quais foram distribuídas em delineamento inteiramente ao acaso, com quatro tratamentos e seis repetições cada. Foram fornecidas dietas com 0, 2, 4 e 6% de folhas desidratadas e moídas de moringa. A inclusão de moringa nas dietas reduziu quadraticamente o consumo até o nível de 1,2%, aumentou o peso dos ovos com comportamento quadrático até o nível de 3,8% e aumentou linearmente o peso da gema. A coloração da gema se intensificou com maiores inclusões de moringa. O perfil bioquímico sofreu alterações leves, mas não saiu dos padrões normais para codornas. A inclusão de 4% de moringa nas dietas aumentou os níveis de fosfatase alcalina. Já a inclusão de 6% reduziu níveis de colesterol e triglicérides. O comportamento das codornas não se alterou com a inclusão de moringa nas dietas. A inclusão de até 3,83% de Moringa oleifera na dieta de codornas japonesas aumenta a qualidade de ovos sem comprometer o desempenho, o perfil bioquímico, os parâmetros sanguíneos e o comportamento delas.(AU)

Diagnosis and treatment of HEp-2 cells contaminated with mycoplasma / Diagnóstico e tratamento de células HEp-2 contaminadas por micoplasma

Abstract Contamination of primary and cell cultures by mycoplasmas is one of the main economic and biological pitfalls in basic research, diagnosis and manufacture of biotechnological products. It is a common issue which may be difficult to conduct surveillance on. Mycoplasma presence may affect several physiological parameters of the cell, besides being considered an important source of inaccurate and/or non-reproducible scientific results. Each cell type presents characteristical symptoms, mainly morphological, that indicate a contamination by mycoplasma. HEp-2 cells originate from carcinoma of the larynx and are, therefore, part of the respiratory tract, which is one of mycoplasma habitats. Despite the importance these cells in several biological research (evaluation of cell proliferation and migration, apoptosis, antiviral and antitumor compounds), the alterations induced by mycoplasma contamination in HEp-2 cells have not yet been described. Here, we describe the progressive morphological alterations in culture of HEp-2 cells infected with mycoplasma, as well as the-diagnosis of the infection and its treatment. Mycoplasma contamination described within this work led to cytoplasm elongation, cell-to-cell spacing, thin plasma membrane projections, cytoplasmic vacuoles, fusion with neighboring cells, and, finally, cell death. Contamination was detected by fluorescence imaging (DAPI) and PCR reactions. The cultures were treated with BM-Cyclin antibiotic to eliminate contamination. The data presented here will be of relevance to researchers whose investigations involve cell culture, especially respiratory and HEp-2 cells.

Resumo A contaminação de culturas primárias e celulares por micoplasmas é uma das principais armadilhas econômicas e biológicas da pesquisa básica, diagnóstico e fabricação de produtos biotecnológicos. Trata-se de uma contaminação rotineira, mas de difícil acompanhamento. A presença de micoplasma pode afetar vários parâmetros fisiológicos da célula, além de ser considerada uma importante fonte de resultados científicos imprecisos e/ou não reprodutíveis. Cada tipo de célula apresenta sintomas característicos, principalmente morfológicos, que indicam uma contaminação por micoplasma. As células HEp-2 são originárias do carcinoma da laringe e, portanto, fazem parte do trato respiratório, um dos habitats do micoplasma. Apesar da importância destas células em diversas pesquisas biológicas (avaliação da proliferação e migração celular, apoptose, compostos antivirais e antitumorais), as alterações decorrentes da contaminação por micoplasma nestas células ainda não foi descrita. Aqui, descrevemos as alterações morfológicas progressivas na cultura de células HEp-2 infectadas por micoplasma, bem como o diagnóstico da infecção e seu tratamento. A contaminação por micoplasma descrita neste trabalho resultou em alongamento citoplasmático, espaçamento entre células, projeções delgadas da membrana plasmática, vacúolos citoplasmáticos, fusão de células vizinhas e, finalmente, morte celular. A contaminação foi detectada por imagens de fluorescência (DAPI) e reações de PCR. As culturas foram tratadas com antibiótico BM-Cyclin para eliminar a contaminação. Os dados aqui apresentados serão de relevância para pesquisadores cujas investigações envolvem cultura celular, principalmente células respiratórias e HEp-2.

Diagnosis and treatment of HEp-2 cells contaminated with mycoplasma.

Contamination of primary and cell cultures by mycoplasmas is one of the main economic and biological pitfalls in basic research, diagnosis and manufacture of biotechnological products. It is a common issue which may be difficult to conduct surveillance on. Mycoplasma presence may affect several physiological parameters of the cell, besides being considered an important source of inaccurate and/or non-reproducible scientific results. Each cell type presents characteristical symptoms, mainly morphological, that indicate a contamination by mycoplasma. HEp-2 cells originate from carcinoma of the larynx and are, therefore, part of the respiratory tract, which is one of mycoplasma habitats. Despite the importance these cells in several biological research (evaluation of cell proliferation and migration, apoptosis, antiviral and antitumor compounds), the alterations induced by mycoplasma contamination in HEp-2 cells have not yet been described. Here, we describe the progressive morphological alterations in culture of HEp-2 cells infected with mycoplasma, as well as the-diagnosis of the infection and its treatment. Mycoplasma contamination described within this work led to cytoplasm elongation, cell-to-cell spacing, thin plasma membrane projections, cytoplasmic vacuoles, fusion with neighboring cells, and, finally, cell death. Contamination was detected by fluorescence imaging (DAPI) and PCR reactions. The cultures were treated with BM-Cyclin antibiotic to eliminate contamination. The data presented here will be of relevance to researchers whose investigations involve cell culture, especially respiratory and HEp-2 cells.

Causes and Pathology of Equine Pneumonia and Pleuritis in Southern Brazil.

We document the causes and pathological findings in 50 cases of equine pneumonia and pleuritis in Southern Brazil. Suppurative (17/50), pyogranulomatous (14/50), aspiration (5/50), mycotic (4/50), bronchointerstitial (3/50), embolic (3/50) and eosinophilic granulomatous pneumonia (1/50) and pleuritis (3/50) were the main conditions identified. Streptococcus spp. were identified in 11 cases of suppurative pneumonia. Suppurative pneumonia was further divided into acute (8/17), subacute (6/17) and chronic (3/17) based on the morphological pattern of lesions. Rhodococcus equi was identified in all cases of pyogranulomatous pneumonia and exclusively affected young foals.

The MLL recombinome of acute leukemias in 2017.

Chromosomal rearrangements of the human MLL/KMT2A gene are associated with infant, pediatric, adult and therapy-induced acute leukemias. Here we present the data obtained from 2345 acute leukemia patients. Genomic breakpoints within the MLL gene and the involved translocation partner genes (TPGs) were determined and 11 novel TPGs were identified. Thus, a total of 135 different MLL rearrangements have been identified so far, of which 94 TPGs are now characterized at the molecular level. In all, 35 out of these 94 TPGs occur recurrently, but only 9 specific gene fusions account for more than 90% of all illegitimate recombinations of the MLL gene. We observed an age-dependent breakpoint shift with breakpoints localizing within MLL intron 11 associated with acute lymphoblastic leukemia and younger patients, while breakpoints in MLL intron 9 predominate in AML or older patients. The molecular characterization of MLL breakpoints suggests different etiologies in the different age groups and allows the correlation of functional domains of the MLL gene with clinical outcome. This study provides a comprehensive analysis of the MLL recombinome in acute leukemia and demonstrates that the establishment of patient-specific chromosomal fusion sites allows the design of specific PCR primers for minimal residual disease analyses for all patients.

Diagnosis and treatment of HEp-2 cells contaminated with mycoplasma

Abstract Contamination of primary and cell cultures by mycoplasmas is one of the main economic and biological pitfalls in basic research, diagnosis and manufacture of biotechnological products. It is a common issue which may be difficult to conduct surveillance on. Mycoplasma presence may affect several physiological parameters of the cell, besides being considered an important source of inaccurate and/or non-reproducible scientific results. Each cell type presents characteristical symptoms, mainly morphological, that indicate a contamination by mycoplasma. HEp-2 cells originate from carcinoma of the larynx and are, therefore, part of the respiratory tract, which is one of mycoplasma habitats. Despite the importance these cells in several biological research (evaluation of cell proliferation and migration, apoptosis, antiviral and antitumor compounds), the alterations induced by mycoplasma contamination in HEp-2 cells have not yet been described. Here, we describe the progressive morphological alterations in culture of HEp-2 cells infected with mycoplasma, as well as the-diagnosis of the infection and its treatment. Mycoplasma contamination described within this work led to cytoplasm elongation, cell-to-cell spacing, thin plasma membrane projections, cytoplasmic vacuoles, fusion with neighboring cells, and, finally, cell death. Contamination was detected by fluorescence imaging (DAPI) and PCR reactions. The cultures were treated with BM-Cyclin antibiotic to eliminate contamination. The data presented here will be of relevance to researchers whose investigations involve cell culture, especially respiratory and HEp-2 cells.

Resumo A contaminação de culturas primárias e celulares por micoplasmas é uma das principais armadilhas econômicas e biológicas da pesquisa básica, diagnóstico e fabricação de produtos biotecnológicos. Trata-se de uma contaminação rotineira, mas de difícil acompanhamento. A presença de micoplasma pode afetar vários parâmetros fisiológicos da célula, além de ser considerada uma importante fonte de resultados científicos imprecisos e/ou não reprodutíveis. Cada tipo de célula apresenta sintomas característicos, principalmente morfológicos, que indicam uma contaminação por micoplasma. As células HEp-2 são originárias do carcinoma da laringe e, portanto, fazem parte do trato respiratório, um dos habitats do micoplasma. Apesar da importância destas células em diversas pesquisas biológicas (avaliação da proliferação e migração celular, apoptose, compostos antivirais e antitumorais), as alterações decorrentes da contaminação por micoplasma nestas células ainda não foi descrita. Aqui, descrevemos as alterações morfológicas progressivas na cultura de células HEp-2 infectadas por micoplasma, bem como o diagnóstico da infecção e seu tratamento. A contaminação por micoplasma descrita neste trabalho resultou em alongamento citoplasmático, espaçamento entre células, projeções delgadas da membrana plasmática, vacúolos citoplasmáticos, fusão de células vizinhas e, finalmente, morte celular. A contaminação foi detectada por imagens de fluorescência (DAPI) e reações de PCR. As culturas foram tratadas com antibiótico BM-Cyclin para eliminar a contaminação. Os dados aqui apresentados serão de relevância para pesquisadores cujas investigações envolvem cultura celular, principalmente células respiratórias e HEp-2.

Tratamento de efluentes de abatedouro de frangos por meio de reator UASB seguido de filtro anaeróbio / Treatment of waste water poultry slaughter house through UASB reactor followed by anaerobic filter

O Brasil é um dos maiores produtores de aves do mundo e se destaca na exportação de carne de frango, que tem papel relevante na economia do país. Como resultado do crescimento da indústria de processamento do frango, há também um significativo aumento na geração de efluentes. Uma alternativa interessante para o tratamento desses resíduos são os reatores anaeróbios. Este trabalho teve como objetivo avaliar o desempenho do sistema reator UASB-Filtro anaeróbio no tratamento de efluentes de abate de frangos. O estudo foi desenvolvido em um abatedouro avícola, situado na região metropolitana de Belo Horizonte, Minas Gerais. O sistema de tratamento foi composto por peneira estática, caixa de gordura, reator UASB e filtro anaeróbio. A vazão média diária de efluentes do abatedouro foi de 30m³/dia. A carga orgânica aplicada no reator UASB foi de 0,71kg DQO/m³.d. O monitoramento do sistema foi realizado por meio de análises físico-químicas do afluente e dos efluentes da caixa de gordura, reator UASB e filtro anaeróbio. Os parâmetros avaliados foram: temperatura, pH, demanda química de oxigênio (DQO), demanda bioquímica de oxigênio (DBO), sólidos suspensos totais (SST), e sólidos suspensos voláteis (SSV). Os valores efluentes médios encontrados para DBO, DQO e SST foram 106, 290 e 56mg/L, respectivamente. A eficiência de remoção do sistema para DBO variou de 83 a 99%, e para DQO de 50 a 98%. A eficiência de remoção média de SST e SSV foi de 92%. Conclui-se que o sistema reator UASB seguido de filtro anaeróbio apresentou alto desempenho na remoção de sólidos e carga orgânica. Dessa forma, esses reatores tornam-se uma alternativa viável para o tratamento de águas residuárias de frigoríficos, já que oferecem bons resultados de remoção de poluentes.

Brazil is one of the largest poultry producers in the world and stands out in exportation of chicken meat, which plays an important role in the economy. As a result of the growth of the poultry processing industry, there is also a significant increase in the generation of wastewater. An interesting alternative for the treatment of such wastes is anaerobic. This study aimed to evaluate the performance of the system UASB reactor-anaerobic filter on wastewater treatment of slaughtering chickens. The study was conducted in a poultry slaughterhouse located in the metropolitan region of Belo Horizonte, Minas Gerais. The treatment system was composed of static sieve, grease trap, UASB reactor and anaerobic filter. The daily average flow of effluent from the abattoir was 30m³/day. The organic load applied to the UASB reactor was 0.71kg COD/m³.d. The monitoring system was performed by means of physico-chemical analysis of the influent and effluent of fat, UASB reactor and anaerobic filter. The parameters evaluated were: temperature, pH, chemical oxygen demand (COD), biochemical oxygen demand (BOD), total suspended solids (SS) and volatile suspended solids (VSS). The mean values for effluent BOD, COD and SS were 106, 290 and 56mg/L, respectively. The removal efficiency of BOD to the system ranged 83-99% for COD and 50-98%. The average removal efficiency of SS and SSV were 92%. We conclude that the system UASB reactor followed by anaerobic filter showed high performance in removing solids and organic load. Thus, these reactors become a viable alternative for treating wastewater slaughterhouses, offering good results in removing pollutants.

Supervisão profissional em serviço social: uma experiência de supervisão externa a coordenadores de serviços domiciliários para pessoas idosas / Professional supervision in social work: an external supervisory experience the home care service coordinators for elderly

O objetivo deste artigo é apresentar um projecto de supervisão profissional desenvolvido por assistentes sociais, docentes do departamento de Serviço Social de uma universidade, denominados de supervisores externos, a assistentes sociais, profissionais responsáveis pela coordenação dos serviços domiciliários para as pessoas idosas, de uma organização sem fins lucrativos. Esta organização desenvolve atividades de ação social na cidade de Lisboa e estabeleceu uma parceria com o departamento de Serviço Social para melhorar as práticas profissionais em serviços de apoio domiciliário para pessoas idosas. No âmbito do relato desta experiência, evidenciamos o projecto de supervisão profissional externa tendo presente alguns casos analisados e considerando o modo como este processo pode potenciar o desenvolvimento pessoal e profissional dos assistentes sociais.

The purpose of this article is to present a professional supervision project developed by social workers, teachers in a Social work Department in a University, called external supervisors to social workers, to professionals responsible for coordinating home care services for the older people in a nonprofit organization. This organization develops social action activities in Lisbon and as established a partnership with the Department of Social work to improve professional practices in home care services for older people. Under this experience, we showed the professional supervision project, taking in to account some cases analyzed, considering the way they can enhance the personal and professional development of social workers.

Polychlorinated biphenyls (PCBs) and p,p'-dichlorodiphenyldichloroethylene (DDE) concentrations in maternal and umbilical cord serum in a human cohort from South Portugal.

Organochlorine compounds as polychlorinated biphenyls (PCBs) and pp'-dichlorodiphenyldichloroethylene (pp'DDE) are ubiquitous, resistant to degradation and lipophilic compounds, commonly found in the general population. Prenatal exposure to these compounds has been associated to adverse developmental effects. Levels of PCBs and pp'DDE were investigated in maternal and umbilical cord serum of 68 women/newborns pairs from Algarve, South Portugal. Mean sum PCBs congeners and pp'DDE concentrations were 1.62 ± 0.39 ng mL(-1)and 1.11 ± 0.69 ng mL(-1)-for maternal and 1.45 ± 0.25 ng mL(-1) and 0.85 ± 0.50 ng mL(-1)for cord serum, respectively. Congeners distribution pattern followed the order PCB 153>180>130, both for maternal and cord serum). Significant correlations (p<0.05) were found between maternal and cord serum concentrations. The umbilical cord/maternal serum ratio were 0.90 and 0.76, respectively for ΣPCB and pp'DDE Multivariate analyses relate women in urban centers with higher PCB levels, while higher pp'DDE relates to older primiparous women that live at rural areas. PCBs were also correlated to more portions of fat rich foods, while pp'DDE is associated to higher ingestion of vegetables and fruits. Smoking habits only correlated to maternal PCB. The present work provides, to our knowledge and for the first time, baseline human-biomonitoring data and establishes background ranges of PCB and pp'DDE levels in the maternal and umbilical cord serum in the Algarve region. These compounds exhibited the capacity to pass the placenta barrier and target the fetus. Even in non-industrialized areas, and in non-intensive agriculture areas, like the Southern Portugal, there is a need to take measures to eliminate or minimize the risk of organochlorine exposure during pregnancy.

Thoracic splenosis as a differential diagnosis of juxtapleural nodules.

Thoracic splenosis is rare and consists of ectopic implantation of splenic tissue into the chest after concomitant thoracic and abdominal trauma with diaphragm injury. It occurs in about 18% of cases of splenic ruptures. In almost all cases, diagnosis is given incidentally once patients are usually asymptomatic. Thoracic splenosis should be considered as a differential diagnosis in all patients with history of trauma presenting with juxtapleural nodules in chest computed tomography. However, malignant conditions should be ruled out firstly. Biopsy is not essential for the diagnosis once nuclear medicine can confirm splenosis in patients with pertinent history of trauma and suggestive tomographic image. We present a typical case of thoracic splenosis whose diagnosis was made by nuclear medicine and no invasive procedures were required.

Quantitative computed tomography of the liver in dogs submitted to prednisone therapy / Tomografia computadorizada quantitativa do fígado de cães submetidos à terapia com prednisona

The aim of this study is to evaluate the possible alterations in radiodensity of the hepatic parenchyma and spleen resulting from the corticotherapy with prednisone, correlating these findings with serum biochemical hepatic transaminase activity. A clinical trial was performed on eight healthy dogs with the intent of evaluating possible hepatic alterations after prednisone therapy using quantitative computed tomography. All animals received prednisone orally at a dose of 2mg/kg of weight for 30 days. The radiodensity of the liver and spleen was determined before and after the period that the medication was administered. An increase in the radiodensity of the hepatic parenchyma between the initial and final experimental stages was observed. In the present study the existence of early hepatic alterations were verified, which are compatible with glycogen storage.

O objetivo deste estudo é avaliar as possíveis alterações na radiodensidade do parênquima hepático e esplênico decorrentes da corticoterapia com prednisona, correlacionando estes achados com níveis séricos de transaminase bioquímicas hepáticas. Um ensaio clínico foi realizado em oito cães sadios com o objetivo de avaliar possíveis alterações hepáticas após a terapia com prednisona por meio de tomografia computadorizada quantitativa. Todos os animais receberam prednisona por via oral na dose de 2mg / kg de peso por 30 dias. A radiodensidade do fígado e do baço foi determinada antes e após o período em que a medicação foi administrada. Foi observado aumento da radiodensidade do parênquima hepático entre os estágios iniciais e finais do experimento. No presente estudo, foi verificada a existência de alterações hepáticas precoces, que são compatíveis com o acúmulo de glicogênio.

Assessment of essential and nonessential metals and different metal exposure biomarkers in the human placenta in a population from the south of Portugal.

The general population is exposed to metals as trace amounts of metallic compounds are present in air, water, and food. Information on background exposures and biomarker concentrations of environmental chemicals in the general Portuguese population is limited. Therefore, the purpose of this study was to determine the levels of important nonessential metals with recognized toxicity cadmium (Cd) and lead (Pb) and essential metals copper (Cu), nickel (Ni), chromium (Cr), and zinc (Zn) in placentas of mothers living in south Portugal (Algarve). Due to the difficulty in establishing the effects of chemicals in a complex and variable environment, this study also aimed to examine the response of biomarkers, such as biochemical changes that occurs at subcellular levels in the presence of contaminants. The investigated biomarkers in placentas indicative of metal exposure or damage included the metallothioneins (MT), delta-aminolevulinic acid dehydratase (ALAD) (specific for Pb), and lipid peroxidation (LPO) as an index of oxidative stress damage. Moreover, HJ-BIPLOT was applied in order to identify and categorize mothers vulnerable to environmental contamination in this region. Metal concentrations in the placenta were not excessive but within the range found in most European studies. In general, the biomarkers MT and LPO were positively correlated with metal levels, while with ALAD the opposite occurred, indicating the selected battery of biomarkers were suitable to study the effects of metals on human placenta. Further, the application of multivariate analysis with HJ-BIPLOT showed that most significant factors contributing to maternal and fetal exposures via placenta were dietary and smoking habits.