RESUMEN
Islet transplantation represents a therapeutic option for type 1 diabetes (T1D). Long-term viability of transplanted islets requires improvement. Mesenchymal stromal cells (MSCs) have been proposed as adjuvants for islet transplantation facilitating grafting and functionality. Stem cell aggregation provides physiological interactions between cells and enhances the in situ concentration of modulators of inflammation and immunity. We established a hanging-drop culture of adult human skin fibroblast-like cells as spheroids, and skin spheroid-derived cells (SphCs) were characterized. We assessed the potential of SphCs in improving islet functionality by cotransplantation with a marginal mass of allogeneic islets in an experimental diabetic mouse model and characterized the secretome of SphCs by mass spectrometry-based proteomics. SphCs were characterized as multipotent progenitors and their coculture with anti-CD3 stimulated mouse splenocytes decreased CD4+ T cell proliferation with skewed cytokine secretion through an increase in the Th2/Th1 ratio profile. SphCs-conditioned media attenuated apoptosis of islets induced by cytokine challenge in vitro and importantly, intratesticular SphCs administration did not show tumorigenicity in immune-deficient mice. Moreover, SphCs improved glycemic control when cotransplanted with a marginal mass of allogeneic islets in a diabetic mouse model without pharmacological immunosuppression. SphCs' protein secretome differed from its paired skin fibroblast-like counterpart in containing 70% of up- and downregulated proteins and biological processes that overall positively influenced islets such as cytoprotection, cellular stress, metabolism, and survival. In summary, SphCs improved the performance of transplanted allogeneic islets in an experimental T1D model, without pharmacological immunosuppression. Future research is warranted to identify SphCs-secreted factors responsible for islets' endurance.
Asunto(s)
Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 1 , Trasplante de Células Madre Hematopoyéticas , Trasplante de Islotes Pancreáticos , Islotes Pancreáticos , Humanos , Ratones , Animales , Adulto , Islotes Pancreáticos/metabolismo , Diabetes Mellitus Tipo 1/terapia , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Experimental/terapia , Diabetes Mellitus Experimental/metabolismo , Citocinas/metabolismoRESUMEN
Islet transplantation represents a therapeutic option for type 1 diabetes (T1D). Long-term viability of transplanted islets requires improvement. Mesenchymal stromal cells (MSCs) have been proposed as adjuvants for islet transplantation facilitating grafting and functionality. Stem cell aggregation provides physiological interactions between cells and enhances the in situ concentration of modulators of inflammation and immunity. We established a hanging-drop culture of adult human skin fibroblast-like cells as spheroids, and skin spheroid-derived cells (SphCs) were characterized. We assessed the potential of SphCs in improving islet functionality by cotransplantation with a marginal mass of allogeneic islets in an experimental diabetic mouse model and characterized the secretome of SphCs by mass spectrometry-based proteomics. SphCs were characterized as multipotent progenitors and their coculture with anti-CD3 stimulated mouse splenocytes decreased CD4+ T cell proliferation with skewed cytokine secretion through an increase in the Th2/Th1 ratio profile. SphCs-conditioned media attenuated apoptosis of islets induced by cytokine challenge in vitro and importantly, intratesticular SphCs administration did not show tumorigenicity in immune-deficient mice. Moreover, SphCs improved glycemic control when cotransplanted with a marginal mass of allogeneic islets in a diabetic mouse model without pharmacological immunosuppression. SphCs' protein secretome differed from its paired skin fibroblast-like counterpart in containing 70% of up- and downregulated proteins and biological processes that overall positively influenced islets such as cytoprotection, cellular stress, metabolism, and survival. In summary, SphCs improved the performance of transplanted allogeneic islets in an experimental T1D model, without pharmacological immunosuppression. Future research is warranted to identify SphCs-secreted factors responsible for islets' endurance.
RESUMEN
The scientific publication landscape is changing quickly, with an enormous increase in options and models. Articles can be published in a complex variety of journals that differ in their presentation format (online-only or in-print), editorial organizations that maintain them (commercial and/or society-based), editorial handling (academic or professional editors), editorial board composition (academic or professional), payment options to cover editorial costs (open access or pay-to-read), indexation, visibility, branding, and other aspects. Additionally, online submissions of non-revised versions of manuscripts prior to seeking publication in a peer-reviewed journal (a practice known as pre-printing) are a growing trend in biological sciences. In this changing landscape, researchers in biochemistry and molecular biology must re-think their priorities in terms of scientific output dissemination. The evaluation processes and institutional funding for scientific publications should also be revised accordingly. This article presents the results of discussions within the Department of Biochemistry, University of São Paulo, on this subject.
Asunto(s)
Bioquímica , Biología Molecular , Publicaciones Periódicas como Asunto/estadística & datos numéricos , Edición/tendencias , Investigación , Brasil , Humanos , Publicaciones Periódicas como Asunto/normas , Publicaciones Periódicas como Asunto/tendenciasRESUMEN
The scientific publication landscape is changing quickly, with an enormous increase in options and models. Articles can be published in a complex variety of journals that differ in their presentation format (online-only or in-print), editorial organizations that maintain them (commercial and/or society-based), editorial handling (academic or professional editors), editorial board composition (academic or professional), payment options to cover editorial costs (open access or pay-to-read), indexation, visibility, branding, and other aspects. Additionally, online submissions of non-revised versions of manuscripts prior to seeking publication in a peer-reviewed journal (a practice known as pre-printing) are a growing trend in biological sciences. In this changing landscape, researchers in biochemistry and molecular biology must re-think their priorities in terms of scientific output dissemination. The evaluation processes and institutional funding for scientific publications should also be revised accordingly. This article presents the results of discussions within the Department of Biochemistry, University of São Paulo, on this subject.
Asunto(s)
Humanos , Publicaciones Periódicas como Asunto/estadística & datos numéricos , Edición/tendencias , Investigación , Bioquímica , Biología Molecular , Publicaciones Periódicas como Asunto/normas , Publicaciones Periódicas como Asunto/tendencias , BrasilAsunto(s)
Actinobacteria/aislamiento & purificación , Bacteriemia/microbiología , Glomerulonefritis/etiología , Trasplante de Corazón/efectos adversos , Terapia de Inmunosupresión/efectos adversos , Antiinfecciosos/administración & dosificación , Antiinfecciosos/uso terapéutico , Bacteriemia/complicaciones , Bacteriemia/tratamiento farmacológico , Cultivo de Sangre , Quimioterapia Combinada , Glomerulonefritis/inmunología , Glomerulonefritis/microbiología , Glomerulonefritis/terapia , Humanos , Terapia de Inmunosupresión/métodos , Inmunosupresores/administración & dosificación , Inmunosupresores/uso terapéutico , Masculino , Persona de Mediana Edad , Daño por Reperfusión Miocárdica/cirugía , Diálisis RenalRESUMEN
Type 1 diabetes mellitus (T1DM) results from death of insulin-secreting ß cells mediated by self-immune cells, and the consequent inability of the body to maintain insulin levels for appropriate glucose homeostasis. Probably initiated by environmental factors, this disease takes place in genetically predisposed individuals. Given the autoimmune nature of T1DM, therapeutics targeting immune cells involved in disease progress have been explored over the last decade. Several high-cost trials have been attempted to prevent and/or reverse T1DM. Although a definitive solution to cure T1DM is not yet available, a large amount of information about its nature and development has contributed greatly to both the improvement of patient's health care and design of new treatments. In this study, we discuss the role of different types of immune cells involved in T1DM pathogenesis and their therapeutic potential as targets and/or modified tools to treat patients. Recently, encouraging results and new approaches to sustain remnant ß cell mass and to increase ß cell proliferation by different cell-based means have emerged. Results coming from ongoing clinical trials employing cell therapy designed to arrest T1DM will probably proliferate in the next few years. Strategies under consideration include infusion of several types of stem cells, dendritic cells and regulatory T cells, either manipulated genetically ex vivo or non-manipulated. Their use in combination approaches is another therapeutic alternative. Cell-based interventions, without undesirable side effects, directed to block the uncontrollable autoimmune response may become a clinical reality in the next few years for the treatment of patients with T1DM.
Asunto(s)
Autoinmunidad/inmunología , Células Dendríticas/inmunología , Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 1/terapia , Células Secretoras de Insulina/inmunología , Trasplante de Células Madre/métodos , Linfocitos T Reguladores/inmunología , Animales , Ensayos Clínicos como Asunto , Células Dendríticas/trasplante , Modelos Animales de Enfermedad , Humanos , Insulina/uso terapéutico , Células Secretoras de Insulina/efectos de los fármacos , Ratones , Ratones Endogámicos NOD , Trasplante de Células Madre/tendencias , Linfocitos T Reguladores/trasplanteRESUMEN
Haemodialysis patients have acquired immunity disturbances, co-morbidities and a vascular access, factors predisposing them to infection and bacteraemia. Clostridium perfringens is an anaerobic bacterium potentially causing severe infections, including rarely septic arthritis. We report the first case of Clostridium perfringens septic arthritis in a haemodialysis patient and suggest a haematogenous spread. After rapid joint lavage combined with appropriate anti-microbial therapy, the patient recovered.
Asunto(s)
Artritis Infecciosa/diagnóstico , Artritis Infecciosa/microbiología , Clostridium perfringens , Nefropatías Diabéticas/complicaciones , Nefropatías Diabéticas/terapia , Articulación de la Cadera , Diálisis Renal , Adulto , Artritis Infecciosa/terapia , Nefropatías Diabéticas/microbiología , Humanos , MasculinoRESUMEN
During the past few years, Epithelial-Mesenchymal Transition (EMT) has emerged as one of the most hot spots in clinical research. Its existence in human tumors can form the basis for explaining characteristics of cancer progression and metastasis, as well as certain cases of drug resistance and relapses after treatment. These cellular responses are tightly regulated by intracellular signaling pathways evoked by humoral factors that include growth factors, chemokines and cytokines. Indeed, several gene regulatory programs known to promote EMT during development have recently been discovered to play key roles in cancer progression. A deeper understanding of the cellular and molecular basis of these different programs should aid in both the development of better diagnosis methods, as well as of specific treatments for invasive cancer. In this review we set out to summarize recent novel insights into the molecular players underlying EMT and its relation with cancer progression and metastasis.
Asunto(s)
Progresión de la Enfermedad , Transición Epitelial-Mesenquimal , Neoplasias/patología , Animales , Transición Epitelial-Mesenquimal/inmunología , Humanos , Metaloproteinasas de la Matriz/metabolismo , MicroARNs/genética , Invasividad Neoplásica , Metástasis de la Neoplasia , Neoplasias/enzimología , Neoplasias/genética , Neoplasias/inmunología , Proteínas Tirosina Quinasas Receptoras/metabolismo , Transducción de Señal , Factor de Crecimiento Transformador beta/inmunologíaRESUMEN
AIMS/HYPOTHESIS: Transplantation of pancreatic islets constitutes a promising alternative treatment for type 1 diabetes. However, it is limited by the shortage of organ donors. Previous results from our laboratory have demonstrated beneficial effects of recombinant human prolactin (rhPRL) treatment on beta cell cultures. We therefore investigated the role of rhPRL action in human beta cell survival, focusing on the molecular mechanisms involved in this process. METHODS: Human pancreatic islets were isolated using an automated method. Islet cultures were pre-treated in the absence or presence of rhPRL and then subjected to serum starvation or cytokine treatment. Beta cells were labelled with Newport green and apoptosis was evaluated using flow cytometry analysis. Levels of BCL2 gene family members were studied by quantitative RT-PCR and western blot. Caspase-8, -9 and -3 activity, as well as nitric oxide production, were evaluated by fluorimetric assays. RESULTS: The proportion of apoptotic beta cells was significantly lowered in the presence of rhPRL under both cell death-induced conditions. We also demonstrated that cytoprotection may involve an increase of BCL2/BAX ratio, as well as inhibition of caspase-8, -9 and -3. CONCLUSIONS/INTERPRETATION: Our study provides relevant evidence for a protective effect of lactogens on human beta cell apoptosis. The results also suggest that the improvement of cell survival may involve, at least in part, inhibition of cell death pathways controlled by the BCL2 gene family members. These findings are highly relevant for improvement of the islet isolation procedure and for clinical islet transplantation.
Asunto(s)
Apoptosis/efectos de los fármacos , Células Secretoras de Insulina/citología , Células Secretoras de Insulina/efectos de los fármacos , Prolactina/farmacología , Proteínas Recombinantes/farmacología , Transducción de Señal/efectos de los fármacos , Adulto , Apoptosis/fisiología , Inhibidores de Caspasas , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Células Cultivadas , Citocinas/metabolismo , Diabetes Mellitus Tipo 1/cirugía , Humanos , Células Secretoras de Insulina/metabolismo , Trasplante de Islotes Pancreáticos/métodos , Persona de Mediana Edad , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Transducción de Señal/fisiología , Proteína X Asociada a bcl-2/metabolismoRESUMEN
The combination of Pleth Variability Index (PVI) and passive leg raising (PLR)-induced pulse pressure variation may help to diagnose hypovolemia in spontaneously breathing patients. In 44 subjects, PVI and Pulse Pressure (PP) variation after PLR were measured before and after induced hypovolemia (blood gift or hemodialysis session). PVI values were significantly greater after hemodialysis session or blood gift (22% vs 18%, P = 0.03); in contrast PP variation did not change significantly (7% vs 4%, P = 0.49). The accuracy of these parameters or of their combination to identify the "after hypovolemia induction" period was weak. In spontaneous ventilation, PVI value is greater after induced hypovolemia, whereas PP variation does not change significantly. The combination of PVI and PLR does not improve the accuracy of the detection of induced hypovolemia.
Asunto(s)
Presión Sanguínea , Hipovolemia/diagnóstico , Respiración , Anciano , Algoritmos , Femenino , Humanos , Pierna , Masculino , Persona de Mediana Edad , Diálisis RenalRESUMEN
We describe an HIV1-positive patient under long-term tenofovir treatment who developed a severe, biopsy-proven, acute tubular necrosis with proximal tubule (PT) dysfunction, precipitated by the very recent start of diclofenac, a nonsteroidal antiinflammatory drug (NSAID). Recent studies show that NSAIDs not only alter glomerular filtration but also multidrug resistance protein (MRP) 4-mediated PT secretion of several substrates. Since the patient tolerated tenofovir well for several years prior to diclofenac use, our observation suggests that diclofenac interfered with tenofovir clearance, thereby favoring its nephrotoxicity. NSAIDs should be avoided in patients under tenofovir.
Asunto(s)
Lesión Renal Aguda/inducido químicamente , Adenina/análogos & derivados , Diclofenaco/efectos adversos , Síndrome de Fanconi/inducido químicamente , Anticuerpos Anti-VIH/inmunología , Seropositividad para VIH/tratamiento farmacológico , VIH-1/inmunología , Organofosfonatos/efectos adversos , Lesión Renal Aguda/patología , Adenina/efectos adversos , Adenina/uso terapéutico , Fármacos Anti-VIH/efectos adversos , Fármacos Anti-VIH/uso terapéutico , Antiinflamatorios no Esteroideos/efectos adversos , Antiinflamatorios no Esteroideos/uso terapéutico , Biopsia , Diagnóstico Diferencial , Diclofenaco/uso terapéutico , Sinergismo Farmacológico , Quimioterapia Combinada , Síndrome de Fanconi/patología , Femenino , Seropositividad para VIH/virología , Humanos , Túbulos Renales Proximales/efectos de los fármacos , Túbulos Renales Proximales/patología , Persona de Mediana Edad , Organofosfonatos/uso terapéutico , TenofovirRESUMEN
Paecilomyces lilacinus is a saprophytic mould which rarely causes infection in humans. We report a case of Paecilomyces lilacinus catheter-related fungemia in a chronic hemodialyzed patient. Blood cultures remained positive for 8 weeks. The infection was cured after eventual acceptance by the patient of oral voriconazole treatment for 6 weeks and removal of the tunneled catheter. The literature on Paecilomyces fungemia in humans is reviewed.
Asunto(s)
Antifúngicos/uso terapéutico , Infecciones Relacionadas con Catéteres/tratamiento farmacológico , Infecciones Relacionadas con Catéteres/microbiología , Catéteres de Permanencia/efectos adversos , Fungemia/tratamiento farmacológico , Fungemia/microbiología , Paecilomyces/aislamiento & purificación , Pirimidinas/uso terapéutico , Diálisis Renal , Triazoles/uso terapéutico , Anciano , Femenino , Humanos , VoriconazolRESUMEN
We report moderate renal failure in a 50-year-old man with a history of recent colonoscopy after oral sodium phosphate purgative use. We initially missed the correct diagnosis, but renal biopsy revealed signs of acute phosphate nephropathy. The patient had residual renal impairment at 8-month follow-up. Greater awareness of this complication is needed amongst health care professionals. The preventive strategies are discussed.
Asunto(s)
Catárticos/efectos adversos , Nefrocalcinosis/inducido químicamente , Biopsia , Colonoscopía , Creatinina/sangre , Humanos , Hiperfosfatemia/inducido químicamente , Enfermedad Iatrogénica , Masculino , Persona de Mediana EdadRESUMEN
Ex vivo islet cell culture prior to transplantation appears as an attractive alternative for treatment of type 1 diabetes. Previous results from our laboratory have demonstrated beneficial effects of human prolactin (rhPRL) treatment on human islet primary cultures. In order to probe into the molecular events involved in the intracellular action of rhPRL in these cells, we set out to identify proteins with altered expression levels upon rhPRL cell treatment, using two-dimensional (2D) gel electrophoresis and mass spectrometry (MS). An average of 300 different protein spots were detected, 14 of which were modified upon rhPRL treatment (p<0.01), of which 12 were successfully identified using MS and grouped according to their biological functions. In conclusion, our study provides, for the first time, information about proteins that could be critically involved in PRL's action on human pancreatic islets, and facilitate identification of new and specific targets involved in islet cell function and proliferation.