RESUMO
Glutathione peroxidases (Gpxs) are antioxidant enzymes not studied so far in legume nodules, despite the fact that reactive oxygen species are produced at different steps of the symbiosis. The function of two Gpxs that are highly expressed in nodules of the model legume Lotus japonicus was examined. Gene expression analysis, enzymatic and nitrosylation assays, yeast cell complementation, in situ mRNA hybridization, immunoelectron microscopy, and LjGpx-green fluorescent protein (GFP) fusions were used to characterize the enzymes and to localize each transcript and isoform in nodules. The LjGpx1 and LjGpx3 genes encode thioredoxin-dependent phospholipid hydroperoxidases and are differentially regulated in response to nitric oxide (NO) and hormones. LjGpx1 and LjGpx3 are nitrosylated in vitro or in plants treated with S-nitrosoglutathione (GSNO). Consistent with the modification of the peroxidatic cysteine of LjGpx3, in vitro assays demonstrated that this modification results in enzyme inhibition. The enzymes are highly expressed in the infected zone, but the LjGpx3 mRNA is also detected in the cortex and vascular bundles. LjGpx1 is localized to the plastids and nuclei, and LjGpx3 to the cytosol and endoplasmic reticulum. Based on yeast complementation experiments, both enzymes protect against oxidative stress, salt stress, and membrane damage. It is concluded that both LjGpxs perform major antioxidative functions in nodules, preventing lipid peroxidation and other oxidative processes at different subcellular sites of vascular and infected cells. The enzymes are probably involved in hormone and NO signalling, and may be regulated through nitrosylation of the peroxidatic cysteine essential for catalytic function.
Assuntos
Regulação da Expressão Gênica de Plantas , Glutationa Peroxidase/genética , Lotus/genética , Proteínas de Plantas/genética , Antioxidantes/metabolismo , Glutationa Peroxidase/metabolismo , Lotus/metabolismo , Organismos Geneticamente Modificados/genética , Organismos Geneticamente Modificados/metabolismo , Proteínas de Plantas/metabolismo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Nódulos Radiculares de Plantas/metabolismo , S-Nitrosoglutationa/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismoAssuntos
Bortezomib/administração & dosagem , Glomerulonefrite Membranoproliferativa , Gamopatia Monoclonal de Significância Indeterminada , Glomerulonefrite Membranoproliferativa/tratamento farmacológico , Glomerulonefrite Membranoproliferativa/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Gamopatia Monoclonal de Significância Indeterminada/tratamento farmacológico , Gamopatia Monoclonal de Significância Indeterminada/patologiaRESUMO
AIM: Further to its pivotal role in haemostasis, factor Xa (FXa) promotes effects on the vascular wall. The purpose of the study was to evaluate if FXa modifies the expression level of energy metabolism and oxidative stress-related proteins in femoral arteries obtained from type 2 diabetic patients with end-stage vasculopathy. METHODS: Femoral arteries were obtained from 12 type 2 diabetic patients who underwent leg amputation. Segments from the femoral arteries were incubated in vitro alone and in the presence of 25 nmol l(-1) FXa and 25 nmol l(-1) FXa + 50 nmol l(-1) rivaroxaban. RESULTS: In the femoral arteries, FXa increased triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase isotype 1 expression but decreased pyruvate dehydrogenase expression. These facts were accompanied by an increased content of acetyl-CoA. Aconitase activity was reduced in FXa-incubated femoral arteries as compared with control. Moreover, FXa increased the protein expression level of oxidative stress-related proteins which was accompanied by an increased malonyldialdehyde arterial content. The FXa inhibitor, rivaroxaban, failed to prevent the reduced expression of pyruvate dehydrogenase induced by FXa but reduced acetyl-CoA content and reverted the decreased aconitase activity observed with FXa alone. Rivaroxaban + FXa but not FXa alone increased the expression level of carnitine palmitoyltransferase I and II, two mitochondrial long chain fatty acid transporters. Rivaroxaban also prevented the increased expression of oxidative stress-related proteins induced by FXa alone. CONCLUSIONS: In femoral isolated arteries from type 2 diabetic patients with end-stage vasculopathy, FXa promoted disruption of the aerobic mitochondrial metabolism. Rivaroxaban prevented such effects and even seemed to favour long chain fatty acid transport into mitochondria.
Assuntos
Diabetes Mellitus Tipo 2/metabolismo , Fator Xa/farmacologia , Artéria Femoral/metabolismo , Acetilcoenzima A/análise , Idoso , Carnitina O-Palmitoiltransferase/genética , Angiopatias Diabéticas/metabolismo , Metabolismo Energético , Feminino , Glicólise , Humanos , Masculino , Mitocôndrias/metabolismo , Morfolinas/farmacologia , Estresse Oxidativo , Rivaroxabana , Tiofenos/farmacologiaRESUMO
Legume nodule senescence is a poorly understood process involving a decrease in N(2) fixation and an increase in proteolytic activity. Some physiological changes during nodule aging have been reported, but scarce information is available at the subcellular level. Biochemical, immunological and proteomic approaches were used to provide insight into the effects of aging on the mitochondria and cytosol of nodule host cells. In the mitochondria, the oxidative modification of lipids and proteins was associated with a marked decline in glutathione, a reduced capacity to regenerate ascorbate, and upregulation of alternative oxidase and manganese superoxide dismutase. In the cytosol, there were consistent reductions in the protein concentrations of carbon metabolism enzymes, inhibition of protein synthesis and increase in serine proteinase activity, disorganization of cytoskeleton, and a sharp reduction of cytosolic proteins, but no detectable accumulation of oxidized molecules. We conclude that nodule mitochondria are an early target of oxidative modifications and a likely source of redox signals. Alternative oxidase and manganese superoxide dismutase may play important roles in controlling ROS concentrations and the redox state of mitochondria. The finding that specific methionine residues of a cytosolic glutamine synthetase isoform are sulfoxidized suggests a regulatory role of this enzyme in senescing nodules.
Assuntos
Mitocôndrias/metabolismo , Phaseolus/metabolismo , Regulação da Expressão Gênica de Plantas , Glutamato-Amônia Ligase/metabolismo , Glutationa/análise , Glutationa/metabolismo , Metabolismo dos Lipídeos , Proteínas Mitocondriais/metabolismo , Proteínas Mitocondriais/fisiologia , Fixação de Nitrogênio , Oxirredução , Oxirredutases/metabolismo , Oxirredutases/fisiologia , Phaseolus/enzimologia , Proteínas de Plantas/metabolismo , Proteínas de Plantas/fisiologia , Isoformas de Proteínas/metabolismo , Processamento de Proteína Pós-Traducional , Proteoma , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Superóxido Dismutase/fisiologiaRESUMO
Seminal vesicles can be affected by tumours originating in other locations. However, primary tumours of the seminal vesicle are extremely rare, with less than 100 cases reported in literature. Seminal vesicle adenocarcinoma is the most common type, but there are also other malign lesions. Diagnosis is challenging due to the lack of early symptoms and well-defined criteria. These tumours are usually asymptomatic and discovered incidentally during imaging tests or pelvic surgery. Definitive diagnosis requires anatomopathological analysis. Case report of 58-years-old man with schwannoma of the seminal vesicle. We describe the main characteristics of these tumours as well as their therapeutic approach.
RESUMO
BACKGROUND: Adolescents' consumption of tranquilizers, sedatives, and sleeping pills (TSSp) has increased during the last few decades, and TSSp are currently among the substances with the lowest age-of-onset. We characterized current-use patterns of TSSp consumers by age when first taken. METHODS: This study used individualized secondary data retrieved from the 2016 Spanish State Survey on Drug Use in Secondary Education (16-18-year-olds), and included all subjects who reported having taken TSSp at any point, but excluded those who had started during the previous year (nâ=â1502). Logistic regression models were used to obtain adjusted odds ratios (aOR) for associations between early TSSp consumption (<14âyears) and current TSSp use patterns, adjusted for sociodemographic factors. RESULTS: About 17.9% of respondents had taken TSSp (average age-of-onsetâ=â13.7) and 45% of these without a prescription. TSSp consumption at <14âyears was higher for males and nonrepeaters. Having begun to use TSSpâ<â14âyears was associated with both higher probability of consumption in the last month (aORâ=â1.41; 95%CI:1.12-1.77) and daily/almost daily consumption in the last month (aORâ=â1.56; 95CI%:1.16-2.08). CONCLUSIONS: The results of this study show there is a high proportion of 16 to 18 TSSp student consumers - both prescribed and nonprescribed; it also establishes that early onset-of-use is associated with higher levels of intensive use later on.
Assuntos
Medicamentos Indutores do Sono , Transtornos Relacionados ao Uso de Substâncias , Tranquilizantes , Adolescente , Humanos , Hipnóticos e Sedativos/efeitos adversos , Masculino , Medicamentos Indutores do Sono/uso terapêutico , Estudantes , Transtornos Relacionados ao Uso de Substâncias/tratamento farmacológico , Tranquilizantes/uso terapêuticoRESUMO
The exact role of viral replication in patients with severe COVID-19 has not been extensively studied, and it has only been possible to demonstrate the presence of replicative virus for more than 3 months in a few cases using different techniques. Our objective was to study the presence of RNA SARS-CoV-2 in autopsy samples of patients who died from COVID-19 long after the onset of symptoms. Secondary superimposed pulmonary infections present in these patients were also studied. We present an autopsy series of 27 COVID-19 patients with long disease duration, where pulmonary and extrapulmonary samples were obtained. In addition to histopathological analysis, viral genomic RNA (gRNA) and viral subgenomic RNA (sgRNA) were detected using RT-PCR and in situ hybridization, and viral protein was detected using immunohistochemistry. This series includes 26 adults with a median duration of 39 days from onset of symptoms to death (ranging 9-108 days), 92% of them subjected to immunomodulatory therapy, and an infant patient. We detected gRNA in the lung of all but one patient, including those with longer disease duration. SgRNA was detected in 11 out of 17 patients (64.7%) with illness duration up to 6 weeks and in 3 out of 9 patients (33.3%) with more than 6 weeks of disease progression. Viral protein was detected using immunohistochemistry and viral mRNA was detected using in situ hybridization in 3 out of 4 adult patients with illness duration of <2 weeks, but in none of the 23 adult patients with an illness duration of >2 weeks. A remarkable result was the detection of viral protein, gRNA and sgRNA in the lung cells of the pediatric patient after 95 days of illness. Additional pulmonary infections included: 9 acute bronchopneumonia, 2 aspergillosis, 2 cytomegalovirus, and 1 BK virus infection. These results suggest that in severe COVID-19, SARS-CoV-2 could persist for longer periods than expected, especially in immunocompromised populations, contributing to the persistence of chronic lung lesions. Additional infections contribute to the fatal course of the disease.
RESUMO
BACKGROUND: Chagas disease is a parasitic disease endemic to Latin America, but it has become a disease of global concern due to migration flows. Asymptomatic carriers may host the parasite for years, without knowing they are infected. The aim of this study is to assess prevalence of Chagas disease and evaluate the participants' level of knowledge between Latin American migrants attending a community-based screening campaign. METHODS: Three community-based campaigns were performed in Alicante (Spain) in 2016, 2017 and 2018, including educational chats and blood tests for Trypanosoma cruzi serology. Participants completed a questionnaire assessing knowledge about the mechanisms of transmission, disease presentation, diagnosis, and treatment. People seropositive for T. cruzi underwent diagnostic confirmation by two different tests. Results were analyzed by multivariable logistic regression and expressed as adjusted odds ratios (aORs), adjusting for age, sex, and time in Spain. RESULTS: A total of 596 participants were included in the study; 17% were aged under 18 years. Prevalence in adults was 11% [54/496; 95% confidence interval (CI): 8.3-14.5%] versus 0% among children. All but one case were in Bolivians. Diagnosis was independently associated with having been born in Bolivia (aOR: 102, 95% CI: 13-781) and a primary school-level education (aOR: 2.40, 95% CI: 1.14-5.06). Of 54 people diagnosed with Chagas disease (most of whom were asymptomatic), 42 (77.7%) returned to the clinic at least once, and 24 (44.4%) received treatment. Multivariable analysis showed that coming from Argentina (aOR: 13, 95% CI: 1.61-1188) or Bolivia (aOR: 1.90, 95% CI: 1.19-3.39) and having received information about Chagas disease in Spain (aOR: 4.63, 95% CI: 2.54-8.97) were associated with a good level of knowledge on the disease. Having primary level studies (aOR: 0.59, 95% CI: 0.34-0.98) and coming from Ecuador (aOR: 4.63, 95% CI: 2.52-847) were independently associated with a lower level of knowledge. CONCLUSIONS: Community-based interventions are a good strategy for diagnosing neglected diseases such as Chagas disease in non-endemic countries and for identifying and treating infected, asymptomatic individuals.
Assuntos
Doença de Chagas/diagnóstico , Migrantes/estatística & dados numéricos , Trypanosoma cruzi/isolamento & purificação , Adulto , Doença de Chagas/epidemiologia , Serviços de Saúde Comunitária , Pesquisa Participativa Baseada na Comunidade , Estudos Transversais , Diagnóstico Precoce , Humanos , América Latina/etnologia , Programas de Rastreamento , Pessoa de Meia-Idade , Doenças Negligenciadas/epidemiologia , Prevalência , Espanha/epidemiologiaRESUMO
BACKGROUND: Renal biopsy (RB) represents the gold standard for diagnosis of kidney diseases. In this paper we analyse whether the indication of RB and histopathology in patients 65 years or older is different from the other patients. MATERIAL AND METHODS: Retrospective study of 93 native renal biopsies performed in the General Hospital of Segovia in the period 2004-2008. The RB was performed percutaneously under ultrasound guidance in real time, using a 16G automatic needle. RESULTS: Mean age of biopsied patients was 56.89 ± 19 (range 14-89) , and 57% were males. A total of 39RB were performed on people aged 65 years or older. Overall, nephrotic syndrome (NS) is the most common indication of RB, and IgA glomerulonephritis the most common histology. In people ≥ 65 years, acute renal failure (ARF) is the most common indication for RB, and rapidly progressing (crescentic) glomerulonephritis/vasculitis the most detected the diagnosis. When taking age into account, no significant differences in the number of glomeruli obtained by RB or in the number of RB performed on the same patient. CONCLUSIONS: In people 65 years or older, ARF is the main indication of RB and crescentic glomerulonephritis/vasculitis the most frequent diagnosis.
Assuntos
Nefropatias/patologia , Rim/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto JovemRESUMO
BACKGROUND: T2 magnetic resonance imaging (T2MR) is a new method for the diagnosis of invasive candidiasis, although most studies have analyzed its role in patients with candidemia or not infection. CASE REPORT: We present the case of a patient with arteritis and thrombosis of the hepatic graft resulted from an undocumented fungal infection in the explanted liver.T2MR in serum was a suitable diagnostic tool for the diagnosis of the deep-seated invasive candidiasis in the absence of candidemia or the isolation of the yeast in culture. CONCLUSIONS: T2MR allowed the diagnosis of deep-seated invasive candidiasis in an immunodepressed patient without candidemia, even before the onset of symptoms.
Assuntos
Candidíase Invasiva/diagnóstico , Transplante de Fígado , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/microbiologia , Candidemia , Candidíase Invasiva/sangue , Feminino , Humanos , Espectroscopia de Ressonância Magnética/métodos , Pessoa de Meia-Idade , Complicações Pós-Operatórias/sangueRESUMO
We describe the case of a male patient who was diagnosed with acute monoblastic leukemia and received a peripheral stem cell transplantation (PSCT) with peripheral blood hematopoietic progenitors. Because he was in clinical remission with no evidence of chronic graft-versus-host disease (GVHD), immunosuppression was withdrawn, and he developed nephrotic syndrome (NS) months later. A kidney biopsy showed focal segmental glomerulosclerosis (FSGS) as part of the GVHD. Soon after the reintroduction of previous immunosuppressive therapy, we observed a complete remission of the NS.
Assuntos
Glomerulosclerose Segmentar e Focal/complicações , Doença Enxerto-Hospedeiro/complicações , Transplante de Células-Tronco Hematopoéticas , Síndrome Nefrótica/diagnóstico , Adulto , Glomerulosclerose Segmentar e Focal/imunologia , Glomerulosclerose Segmentar e Focal/patologia , Doença Enxerto-Hospedeiro/imunologia , Doença Enxerto-Hospedeiro/patologia , Humanos , Terapia de Imunossupressão , Leucemia Monocítica Aguda/terapia , Masculino , Síndrome Nefrótica/etiologia , Síndrome Nefrótica/terapiaRESUMO
BACKGROUND: Interferon therapy at the injection sites has been related to different cutaneous lesions including erythema and induration as the most frequent ones. While the glycoprotein induces fatigue, fever and is even believed to precipitate autoimmune disorders such as type I diabetes, thyroid disease and systemic lupus erythematosus, a lupus erythematosus-like histologic reaction at the interferon injection site has never been reported. To our knowledge, a microscopic self-resolving lesion mimicking lupus erythematosus at the injection site of interferon has not been described. RESULTS: We report five cases of cutaneous lesions at the inoculation site of interferon with a histopathologic lupus erythematosus-like pattern. Three of them were receiving interferon alfa therapy because of a malignant melanoma and the other two patients were receiving interferon beta-1b for multiple sclerosis. Biopsy specimens taken from different lesions showed similar microscopic findings consisting of dermal mucin deposits and dense lymphocytic infiltrates along hair follicles with hydropic degeneration of follicular basal layer. CONCLUSIONS: Multiple cutaneous lesions related to interferon at the injection site have been reported, but none of them with a histologic lupus-like presentation. In this study we describe five cases in which interferon therapy has induced a resolutive cutaneous lesion mimicking lupus erythematous. This peculiar microscopic pattern has been previously described once before, but interpreted as cutaneous mucinosis.
Assuntos
Toxidermias/etiologia , Interferon-alfa/efeitos adversos , Interferon beta/efeitos adversos , Lúpus Eritematoso Cutâneo/diagnóstico , Pele/efeitos dos fármacos , Adulto , Diagnóstico Diferencial , Toxidermias/patologia , Feminino , Humanos , Injeções Subcutâneas , Interferon alfa-2 , Interferon beta-1b , Masculino , Pessoa de Meia-Idade , Proteínas Recombinantes , Pele/patologiaRESUMO
Ischemia/reperfusion (I/R) leads to Acute Kidney Injury. HIF-1α is a key factor during organ response to I/R. We previously demonstrated that HIF-1α is induced during renal reperfusion, after ischemia. Here we investigate the role of HIF-1α and the HIF-1α dependent mechanisms in renal repair after ischemia. By interference of HIF-1α in a rat model of renal I/R, we observed loss of expression and mis-localization of e-cadherin and induction of α-SMA, MMP-13, TGFß, and collagen I. Moreover, we demonstrate that HIF-1α inhibition promotes renal cell infiltrates by inducing IL-1ß, TNF-α, MCP-1 and VCAM-1, through NFkB activity. In addition, HIF-1α inhibition induced proximal tubule cells proliferation but it did not induce compensatory apoptosis, both in vivo. In vitro, HIF-1α knockdown in HK2 cells subjected to hypoxia/reoxygenation (H/R) promote cell entry into S phase, correlating with in vivo data. HIF-1α interference leads to downregulation of miR-127-3p and induction of its target gene Bcl6 in vivo. Moreover, modulation of miR-127-3p in HK2 cells subjected to H/R results in EMT regulation: miR127-3p inhibition promote loss of e-cadherin and induction of α-SMA and collagen I. In conclusion, HIF-1α induction during reperfusion is a protector mechanism implicated in a normal renal tissue repair after I/R.
Assuntos
Injúria Renal Aguda/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Isquemia/metabolismo , Rim/metabolismo , Traumatismo por Reperfusão/metabolismo , Injúria Renal Aguda/etiologia , Animais , Apoptose , Ciclo Celular , Proliferação de Células , Modelos Animais de Doenças , Fibrose , Mediadores da Inflamação/metabolismo , Isquemia/complicações , Rim/irrigação sanguínea , Rim/patologia , Macrófagos/metabolismo , Masculino , Nefrite/complicações , Nefrite/metabolismo , Ratos Sprague-Dawley , Traumatismo por Reperfusão/complicaçõesAssuntos
Adalimumab/efeitos adversos , Anti-Inflamatórios/efeitos adversos , Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/induzido quimicamente , Doença de Crohn/tratamento farmacológico , Adalimumab/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The digital workflow in dentistry allows for complete digital processing of the restoration starting with the digital impression using an intraoral scanner to until the fabrication of final reconstruction. Recent advances in 3D printing technologies opened new possibilities also for dental technicians through which wax-up and casting procedures in the laboratories could be eliminated. In this clinical report, a technique is described where the pattern was fabricated using additive manufacturing for pressed lithium disilicate onlay restorations. (AU)
O fluxo de trabalho digital em odontologia permite o processamento digital completo da restauração começando com a impressão digital usando um scanner intraoral até a fabricação da peça final. Os avanços recentes nas tecnologias de impressão 3D abriram novas possibilidades também para os técnicos em prótese dentária através dos quais os procedimentos de cera e fundição nos laboratórios poderiam ser eliminados. Neste relato de caso clínico, descreve-se uma técnica onde o padrão foi fabricado usando a fabricação de aditivos para restaurações do tipo onlay em dissilicato de lítio injetadas. (AU)
Assuntos
Humanos , Masculino , Adulto , Impressão Tridimensional , Síndrome de Dente Quebrado/diagnóstico por imagem , Restaurações Intracoronárias/métodosRESUMO
Acute tubular necrosis (ATN) caused by ischemia/reperfusion (I/R) during renal transplantation delays allograft function. Identification of factors that mediate protection and/or epithelium recovery could help to improve graft outcome. We studied the expression, regulation and role of hypoxia inducible factor 1-alpha (HIF-1 α), using in vitro and in vivo experimental models of I/R as well as human post-transplant renal biopsies. We found that HIF-1 α is stabilized in proximal tubule cells during ischemia and unexpectedly in late reperfusion, when oxygen tension is normal. Both inductions lead to gene expression in vitro and in vivo. In vitro interference of HIF-1 α promoted cell death and in vivo interference exacerbated tissue damage and renal dysfunction. In pos-transplant human biopsies, HIF-1 α was expressed only in proximal tubules which exhibited normal renal structure with a significant negative correlation with ATN grade. In summary, using experimental models and human biopsies, we identified a novel HIF-1 α induction during reperfusion with a potential critical role in renal transplant.