Unveiling the mechanisms underlying photothermal efficiency of gold shell-isolated nanoparticles (AuSHINs) on ductal mammary carcinoma cells (BT-474).

Gold nanoparticles are valuable photothermal agents owing to their efficient photothermal conversion, photobleaching resistance, and potential surface functionalization. Herein, we combined bioinspired membranes with in vitro assays to elicit the molecular mechanisms of gold shell-isolated nanoparticles (AuSHINs) on ductal mammary carcinoma cells (BT-474). Langmuir and Langmuir-Schaefer (LS) films were handled to build biomembranes from BT-474 lipid extract. AuSHINs incorporation led to surface pressure-area (π-A) isotherms expansion, increasing membrane flexibility. Fourier-transform infrared spectroscopy (FTIR) of LS multilayers revealed electrostatic AuSHINs interaction with head portions of BT-474 lipid extract, causing lipid chain disorganization. Limited AuSHINs insertion into monolayer contributed to hydroperoxidation of the unsaturated lipids upon irradiation, consistently with the surface area increments of ca. 2.0%. In fact, membrane disruption of irradiated BT-474 cells containing AuSHINs was confirmed by confocal microscopy and LDH leakage, with greater damage at 2.2 × 1013 AuSHINs/mL. Furthermore, the decrease in nuclei dimensions indicates cell death through photoinduced damage.

The use of Artificial Intelligence in predicting Respiratory Syncytial Virus-inhibiting flavonoids.

Human Respiratory Syncytial Virus (hRSV) infection results in death and hospitalization of thousands of people worldwide each year. Unfortunately, there are no vaccines or specific treatments for hRSV infections. Screening hundreds or even thousands of promising molecules is a challenge for science. We integrated biological, structural, and physicochemical properties to train and to apply the concept of artificial intelligence (AI) able to predict flavonoids with potential anti-hRSV activity. During the training and simulation steps, the AI produced results with hit rates of more than 83%. The better AIs were able to predict active or inactive flavonoids against hRSV. In the future, in vitro and/or in vivo evaluations of these flavonoids may accelerate trials for new anti-RSV drugs, reduce hospitalizations, deaths, and morbidity caused by this infection worldwide, and be used as input in these networks to determine which parameter is more important for their decision.

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.

Quercetin inhibits neutrophil extracellular traps release and their cytotoxic effects on A549 cells, as well the release and enzymatic activity of elastase and myeloperoxidase.

Neutrophil extracellular traps (NETs) were first reported as a microbicidal strategy for activated neutrophils. Through an immunologic response against several stimuli, neutrophils release their DNA together with proteins from granules, nucleus, and cytoplasm (e.g., elastase and myeloperoxidase). To date, NETs have been implicated in tissue damage during intense inflammatory processes, mainly when their release is dependent on oxygen radical generation. Flavonoids are antioxidant and anti-inflammatory agents; of these, quercetin is commonly found in our daily diet. Therefore, quercetin could exert some protective activity against tissue damage induced by NETs. In our in vitro assays, quercetin reduced NETs, myeloperoxidase (MPO), and elastase release from neutrophils stimulated with phorbol 12-myristate 13-acetate (PMA). The activity of these enzymes also decreased in the presence of quercetin. Quercetin also reduced the cytotoxic effect of NETs on alveolar cells (A549 cell line). Further, in silico assays indicated favorable interactions between quercetin and NET proteins (MPO and elastase). Overall, our results demonstrate that quercetin decreases deleterious cellular effects of NETs by reducing their release from activated neutrophils, and diminishing the enzymatic activity of MPO and elastase, possibly through direct interaction.

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.

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.

Hipercementosis, una alteración de la estructura dentaria: recopilación bibliográfica / Hypercementosis, an alteration of the dental structure: bibliographic compilation

La hipercementosis se presenta como depósitos de cemento en la raíz de uno o más dientes. Siendo más frecuente en dientes que están sometidos a fuerzas de oclusión, particularmente en pacientes con enfermedad de Paget. El aumento de cemento se depositará en una banda que se encuentra situada alrededor del tercio apical de la raíz, es de ayuda útil realizar una evaluación radiográfi ca antes de una extracción dental (AU)

Hypercementosis occurs as cement deposits in the root of one or more teeth. Being more frequent in teeth that are subjected to occlusion forces or in patients with Paget's disease. The cement increase can be deposited in a band that can be found near the apical third of the root, it is a dental aid that a radiographic evaluation is made before a dental extraction (AU)

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.

Recurrent membranoproliferative glomerulonephritis after second renal graft treated with plasmapheresis and rituximab.

We present a case of a 45-year-old man who suffered from idiopatic membranoproliferative glomerulonephritis (MPGN) in the native kidney that relapsed after his first and second renal grafts. The patient was diagnosed in 1990 with lobular MPGN type I, receiving his first renal graft in 1996. In 2001, a biopsy showed recurrence of MPGN type I (rMPGN). He underwent a second renal graft in 2008. In January 2010, he experienced increased proteinuria and creatinine. Upon electron microscopy of a renal graft biopsy we diagnosed a new rMPGN. At the time of the biopsy, complement levels were normal, although C3 and C4 decreased further. We administered 12 plasmapheresis (PP) sessions and four doses of rituximab. Due to persistent renal impairment, we performed a new biopsy 3 months later, showing less severity of the acute lessions. He received a new cycle of treatment (PP+rituximab). One year later, his renal function was stable with a creatinine ranging between 2 and 2.5 mg/dL and a protein/creatinine ratio less than 1 mg/mg. We concluded that the treatment stopped the disease progression.

Long-term survival of simultaneous pancreas-kidney transplantation: influence of early posttransplantation complications.

BACKGROUND: Complications that develop in the early posttransplantation period after simultaneous pancreas-kidney transplantation (SPKT) can contribute to poor long-term survival of grafts and patients. PATIENTS AND METHODS: We studied 86 SPKTs that were performed between 2000 and 2010 in our hospital, analyzing all complications in the early posttransplantation period and their impact on long-term survival in patients and grafts. RESULTS: The mean age of the patients was 38.77 ± 7.13 years (79.1% male). Of the 86 SPKT patients, 22.1% were on peritoneal dialysis (PD) before transplantation, 68.6% were on hemodialysis (HD), and 9.3% had not received any substitutive renal therapy. The immunosuppressive regimens consisted of induction with basiliximab followed by tacrolimus, mycophenolate mofetil, and steroid therapy. More than 75% of patients experienced an infection in the early posttransplantation period: bacteremia (37.2%), central catheter infection (7%), wound infection (4.7%), urinary tract (14%) and positive abdominal drain culture (45.3%). Approximately one third (31.4%) of patients underwent a reoperation, primarily due to bleeding (21.95%) or infection (19.51%). One fifth of patients (19.8%) experienced an acute rejection episode. The 3-year survival of the pancreas was lower among PD patients (82%) compared with patients who did not undergo dialysis before SPKT (100%). The 5-year survival rate of both grafts was lower among patients who underwent a reoperation than those who did not: pancreas survival rates, were 70% versus 93%, respectively (P = .015) and kidney graft survival rates were 75% versus 96%, respectively (P = .0017). Five-year patient survival rates were also lower among reoperated patients than those who were not (85% vs 97%, respectively), although the difference was not significant (P = .27). CONCLUSIONS: Complications in the early posttransplantation period after SPKT were frequent, increasing morbidity and inpatient stay. One third of our patients underwent a reoperation, which had a negative impact on graft survival.

Impact of recurrent glomerulonephritis on renal graft survival.

BACKGROUND: Glomerular disease causes graft loss in the intermediate and long term, especially recurrent primary renal disease, negatively impacting graft survival. Thus, it must be considered a differential diagnosis in the evaluation of chronic graft dysfunction. METHODS: The objectives of our study were to compare the impacts of primary glomerular disease on graft survival and association with interstitial fibrosis/tubular atrophy (IFTA) or transplant glomerulopathy. We examined the influence of the relapse of glomerulonephritis (GN) on renal graft survival in a retrospective study of 1057 patients undergoing renal transplantations between March 1981 and October 2009. Among this group, 128 patients were diagnosed with pretransplant GN by renal biopsy. We examined graft survival on recurrence compared with IFTA and transplant glomerulopathy using Kaplan-Meier analysis. RESULTS: We analyzed a cohort of 128 patients who were diagnosed with pretransplant GN by renal biopsy, including 28.9% (37) of whom were males. The mean age was 42.04 ± 13.82 years. The most frequent type was immunoglobulin A GN (IgAGN; 31.3%), followed by membranoproliferative GN (MPGN; 28.9%), rapidly progressive GN (RPGN; 16.4%), focal-segmental GN (FSGN; 13.3%), membranous GN (9.4%), and minimal change GN; (0.8%). Among the 16 cases (12.5%) of GN recurrence; MPGN was associated most frequently (n = 10, 28.9%), followed by FSGN (n = 4, 23.5%), RPGN (n = 1, 4.8%), and IgAGN (n = 1, 2.5%). We noted that 11.8% of subjects to be positive for hepatitis C virus; while 3.9% were hepatitis B virus(HBV)-positive. We observed no differences in hepatic serology between patients who experienced recurrence (HBV 6.3% vs hepatitis C virus [HCV] 18.8%) compared with IFTA (HBV 3.1% vs HCV 9.4%). Fifty-one patients (39.8%) were biopsied after transplantation due to impaired renal function: there were recurrences of GN in 12.5% (n = 16), IFTA in 25% (n = 32), and transplant glomerulopathy in 2.3% (n = 3) cases. The average graft survival in our cohort was 8.36 ± 0.59 years. The median patient survival among those who experienced a recurrence was 8.36 ± 1.79 years; 7.19 ± 1.01 years in IFTA patients; and 3.31 ± 0.91 years in patients with transplant glomerulopathy (log-rank P = .06). Upon multivariate analysis, recurrence of GN was not an independent predictor of renal loss. CONCLUSIONS: MPGN was the type of GN that recurred most frequently followed by FSGN. No differences in graft survival were noted between long-term recurrence of GN and other causes of chronic graft dysfunction. The recurrence of primary disease did not worsen the renal graft prognosis versus other causes of chronic graft dysfunction.

Síndrome antisintetasa sin miositis con amiloidosis secundaria AA: una asociación no descrita / No disponible

No disponible

[Complications associated with percutaneous renal biopsy in Spain, 50 years later]. / Complicaciones asociadas a la biopsia renal percutánea. Experiencia en España 50 años después.

BACKGROUND: The renal biopsy is essential for the diagnostic of glomerular disease However, it is an aggressive procedure with risk of complications. OBJECTIVES: The aim of our study was to evaluate the complications directly related to percutaneous renal biopsy procedure in our centre. METHODS: This retrospective study was performed using the data obtained from all patients who underwent percutaneous renal biopsy of the native kidney from January 1992 to December 2008. A semiautomatic 18 G needle biopsy was used until 2004 and thereafter we used a 16 G needle. From January 2009 to January 2010 we prospectively analyzed changes induced by renal biopsy. We analysed age, sex, indication for biopsy, histopathological diagnosis, hypertension, serum creatinine, GFR-MDRD-4, proteinuria, hemoglobin pre and post biopsy. Minor complications were defined as a decrease in hemoglobin levels greater than 1 g/dL. Mayor complications were: need for blood transfusion, surgery, nephrectomy, angiography, embolization, or death. The renal biopsy was performed by the nephrologist with the help of ultrasound. Anticoagulant therapy was removed prior to the biopsy. RESULTS: Total number of renal biopsies were 867. Seven hundred and ninety five renal biopsies were performed between 1992 and 2008. The prospective part of the study included 70 additional biopsies. Considering all patients, the mean age was 46.8 ± 19 and 60.7% were male. There were only six major complications (0.75%). Three of these mayor complications occurred in liver transplanted patients and required vascular embolization or nephrectomy. The remaining 3 major complications were observed in: one patient with liver disease, another patient had trait of hemophilia and a third patient required nephrectomy which after examination demonstrated epithelioid hemangioma. During the prospective analysis the rate of major and minor complications did not change, 1.4 and 2.0 % respectively. Switching from 18 to 16 G biopsy needle did not result in an increase of major complications. CONCLUSIONS: Major complications derived from all renal biopsy during the last 18 years were observed in only 0.75-1.4 %. Major complications occurred mainly in liver transplant patients. The use of 16 G needle provided greater diagnostic yield than the 18 G and it did not cause an increase in complications.

Complicaciones asociadas a la biopsia renalpercutánea. Experiencia en España 50 años después / Complications associated with percutaneous renal biopsy in Spain, 50 years later

Antecedentes: La biopsia renal (BR) es una técnica fundamental en el estudio de las enfermedades renales. Es también el procedimiento más agresivo por su morbimortalidad, por lo cual resulta fundamental conocer sus complicaciones. Objetivos: El objetivo de nuestro estudio fue cuantificar las complicaciones de la BR percutánea en nuestro centro. Métodos: Se realizó un estudio retrospectivo de todos los pacientes a los que se les realizó una BR percutánea de riñón nativo entre enero de 1992 y diciembre de 2008. Hasta el año 2004 usamos una aguja semiautomática de 18 Gauges (G) y desde esa fecha, de 16 G. Se realizó, además, un estudio prospectivo desde enero de 2009 hasta enero de 2010. Se analizaron: edad, sexo, indicación de biopsia, diagnóstico histopatológico, hipertensión arterial (HTA), creatinina sérica, GFR-MRD-4, proteinuria y hemoglobina previa y posterior a la biopsia. Definimos complicaciones menores como: descenso de la hemoglobina mayor de 1 g/dl y como complicaciones mayores la necesidad de transfusión, cirugía, nefrectomía, arteriografía, embolización o muerte. La BR fue realizada por el equipo de nefrología con control ecográfico y retirando el tratamiento antiagregante. Resultados: El número total de biopsias realizadas en los últimos 18 años ha sido de 867. En el estudio retrospectivo, desde enero de 1992 hasta diciembre de 2008, se realizaron 797 biopsias renales. La edad media de los pacientes fue de 46,8 ± 19,1 años y el 60,7% de ellos eran hombres. Sólo observamos seis complicaciones mayores (0,75%). Tres de los pacientes que las presentaron habían sido sometidos a trasplante hepático, presentaron complicaciones hemorrágicas, dos de ellos precisaron embolización y uno nefrectomía. Las tres restantes complicaciones se presentaron en una paciente hepatópata, una afectada de hemofilia y en la tercera se realizó nefrectomía que evidenció hemangiomas epitelioides múltiples. En el estudio prospectivo (enero de 2009-2010) se han realizado 70 biopsias, observándose complicaciones mayores en un 1,4% (un caso) y menores en un 2% (un caso), datos similares a los del estudio retrospectivo. No hubo diferencias en complicaciones mayores entre la aguja de 16 y la de 18 G. Conclusiones: Las complicaciones mayores fueron del 0,75-1,4% y se presentan, sobre todo, en pacientes sometidos a trasplante hepático. Con el empleo de la aguja de 16 G no se observaron más complicaciones mayores y sí una mayor rentabilidad diagnóstica (AU)

Background: The renal biopsy is essential for the diagnostic of glomerular disease However, it is an aggressive procedure with risk of complications. Objectives: The aim of our study was to evaluate the complications directly related to percutaneous renal biopsy procedure in our centre. Methods: This retrospective study was performed using the data obtanined from all patients who underwent percutaneous renal biopsy of the native kidney from January 1992 to December 2008. A semiautomatic 18 G needle biopsy was used until 2004 and thereafter we used a 16 G needle. From January 2009 to January 2010 we prospectively analyzed changes induced by renal biopsy. We analysed age, sex, indication for biopsy, histopathological diagnosis, hypertension, serum creatinine, GFR-MDRD-4, proteinuria, hemoglobin pre and post biopsy. Minor complications were defined as a decrease in hemoglobin levels greater than 1 g/dL. Mayor complications were: need for blood transfusion, surgery, nephrectomy, angiography, embolization, or death. The renal biopsy was performed by the nephrologist with the help of ultrasound. Anticoagulant therapy was removed prior to the biopsy. Results: Total number of renal biopsies were 867. Seven hundred and ninety five renal biopsies were performed between 1992 and 2008. The prospective part of the study included 70 additional biopsies. Considering all patients, the mean age was 46.8±19 and 60.7% were male. There were only six major complications (0.75%). Three of these mayor complications occurred in liver transplanted patients and required vascular embolization or nephrectomy. The remaining 3 major complications were observed in: one patient with liver disease, another patient had trait of hemophilia and a third patient required nephrectomy which after examination demostrated epitheliod hemanangioma. During the prospective analysis the rate of major and minor complications did not change, 1.4 and 2.0 % respectively. Switching from 18 to 16 G biopsy needle did not result in an increase of major complications. Conclusions: Major complications derived from all renal biopsy during the last 18 years were observed in only 0.75-1.4 %. Major complications occurred mainly in liver transplant patients. The use of 16 G needle provided greater diagnostic yield than the 18 G and it did not cause an increase in complications (AU)

Neutrophil haptotaxis induced by mouse MNCF: interactions with extracellular matrix glycoproteins probably contribute to overcoming the anti-inflammatory action of dexamethasone.

OBJECTIVE AND DESIGN: The Macrophage-derived Neutrophil Chemotactic Factor (MNCF) has been characterized as a dexamethasone-resistant neutrophil chemotactic lectin produced by rat macrophages. This study was undertaken to evaluate different MNCF cellular sources and investigate the mechanisms by which MNCF overcomes the anti-inflammatory actions of dexamethasone. MATERIAL AND METHODS: The mouse macrophage-like cell line P388D1 and thioglycollate-elicited mouse macrophages were studied regarding their capacity to release MNCF. Neutrophil migration assays were performed in vivo and in vitro, in either the presence or absence of extracellular matrix glycoproteins (ECM). RESULTS: Mouse and P388D1 macrophages release a lectin that reproduces the activities of rat MNCF. The ability of MNCF to induce neutrophil adhesion and haptotaxis is enhanced through its interaction with laminin and fibronectin. These properties are not inhibited by dexamethasone. CONCLUSIONS: Together, our results suggest that dexamethasone-resistant neutrophil migration induced by MNCF occurs probably because of its interactions with ECM.