Asunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Antagonistas del Receptor de Bradiquinina B2/uso terapéutico , Bradiquinina/análogos & derivados , Tratamiento Farmacológico de COVID-19 , Bradiquinina/uso terapéutico , COVID-19/diagnóstico , COVID-19/fisiopatología , Humanos , Pulmón/efectos de los fármacos , Pulmón/fisiopatología , Masculino , Persona de Mediana Edad , SARS-CoV-2 , Resultado del TratamientoRESUMEN
High LDL-cholesterol (LDL-C) characterizes familial hypercholesterolemia (FH) and familial combined hyperlipidemia (FCH). LDL-apheresis, used in these patients to reduce LDL-C levels, has been shown to also affect HDL levels and composition. We studied LDL-apheresis effects on six FH and nine FCH subjects' serum capacity to modulate cellular cholesterol efflux, an index of HDL functionality, and to load macrophages with cholesterol. Serum cholesterol efflux capacity (CEC) and macrophage cholesterol loading capacity (CLC) were measured before, immediately after, and two days after LDL-apheresis. The procedure reduced total cholesterol (TC), LDL-C, and apoB plasma levels (-69%, -80% and -74%, respectively), parameters only partially restored two days later. HDL-C and apoA-I plasma levels, reduced after LDL-apheresis (-27% and -16%, respectively), were restored to almost normal levels two days later. LDL-apheresis reduced serum aqueous diffusion (AD) CEC, SR-BI-CEC, and ABCA1-CEC. AD and SR-BI were fully restored whereas ABCA1-CEC remained low two days later. Sera immediately and two days after LDL-apheresis had a lower CLC than pre-LDL-apheresis sera. In conclusion, LDL-apheresis transiently reduces HDL-C levels and serum CEC, but it also reduces also serum capacity to deliver cholesterol to macrophages. Despite a potentially negative effect on HDL levels and composition, LDL-apheresis may counteract foam cells formation.
Asunto(s)
Eliminación de Componentes Sanguíneos , LDL-Colesterol/sangre , LDL-Colesterol/metabolismo , Transportador 1 de Casete de Unión a ATP , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 1 , Transportadoras de Casetes de Unión a ATP/metabolismo , Transporte Biológico , Antígenos CD36/metabolismo , Difusión , Femenino , Humanos , Hiperlipoproteinemia Tipo II/sangre , Hiperlipoproteinemia Tipo II/terapia , Macrófagos/metabolismo , Masculino , Persona de Mediana Edad , Factores de Tiempo , Agua/metabolismoRESUMEN
Kidney transplantation in morbidly obese patients can be technically demanding. Furthermore, morbidly obese patients experience a high rate of wound infections and related complications, which mostly result from the longer length and extent of the incision. These complications can be avoided through minimally invasive surgery; however, conventional laparoscopic instruments are unsuitable for the safe performance of a kidney transplant in morbidly obese patients. Herein, we report the first minimally invasive, total robotic kidney transplant in a morbidly obese patient. A left, deceased donor kidney was transplanted into a 29-year-old woman with a body mass index (BMI) of 41 kg/m(2) who had been on hemodialysis for 5 years. The operation was performed intraabdominally using the DaVinci Robotic Surgical System with 4 trocars and a 7 cm midline incision. The operative time was 223 min, and the blood loss was less than 50 cc. The kidney had immediate graft function. No perioperative complications were observed, and the patient was discharged on postoperative day 5 with normal kidney function. Minimally invasive access and robotic technology facilitated the safe performance of a successful kidney transplant in a morbidly obese patient.
Asunto(s)
Trasplante de Riñón/efectos adversos , Obesidad/complicaciones , Adulto , Índice de Masa Corporal , Femenino , Humanos , Laparoscopía , Morbilidad , Robótica , Donantes de Tejidos , Resultado del TratamientoRESUMEN
This study aimed to analyse the sarcolemma of Col6a1-/- fibers in comparison with wild type and mdx fibers, taken as positive control in view of the known structural and functional alterations of their membranes. Structural and mechanical properties were studied in single muscle fibers prepared from FDB muscle using atomic force microscopy (AFM) and conventional electrophysiological techniques to measure ionic conductance and capacitance. While the sarcolemma topography was preserved in both types of dystrophic fibers, membrane elasticity was significantly reduced in Col6a1-/- and increased in mdx fibers. In the membrane of Col6a1-/- fibers ionic conductance was increased likely due to an increased leakage, whereas capacitance was reduced, and the action potential (ap) depolarization rate was reduced. The picture emerging from experiments on fibers in culture was consistent with that obtained on intact freshly dissected muscle. Mdx fibers in culture showed a reduction of both membrane conductance and capacitance. In contrast, in mdx intact FDB muscle resting conductance was increased while resting potential and ap depolarization rate were reduced, likely indicating the presence of a consistent population of severely altered fibers which disappear during the culture preparation.
Asunto(s)
Colágeno Tipo VI/fisiología , Distrofina/fisiología , Fibras Musculares Esqueléticas/fisiología , Distrofias Musculares/fisiopatología , Sarcolema/fisiología , Potenciales de Acción/fisiología , Análisis de Varianza , Animales , Supervivencia Celular/fisiología , Colágeno Tipo VI/biosíntesis , Colágeno Tipo VI/genética , Modelos Animales de Enfermedad , Distrofina/genética , Electrofisiología/métodos , Potenciales de la Membrana/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos mdx , Microscopía de Fuerza Atómica , Fibras Musculares Esqueléticas/ultraestructura , Sarcolema/ultraestructura , Técnicas de Cultivo de TejidosRESUMEN
Cerato-ulmin (CU) is a fungal toxin class II hydrophobin, involved in Dutch elm disease. The formation of hydrophobin films at the air-water interface is a key mechanism which plays a role of paramount importance at different stages of the fungal development. We present a study on the precursor stages of growth towards the self-assembly aggregation film of CU. Atomic force microscopy images of CU dropped on mica substrates indicate that the system self-organizes in almost one-dimensional pearl-necklace-like chains, which subsequently collapse and possibly merge to form extended and rather compact planar films. We propose and verify a simple model to describe the self-aggregation mechanism in terms of progressive thickening of the pearl chains due to the successive merging and collapse of the elementary constitutive units.
Asunto(s)
Proteínas Fúngicas/metabolismo , Micotoxinas/metabolismo , Ophiostoma/química , Agua/química , Aire/análisis , Animales , Microscopía de Fuerza Atómica/métodos , Pinctada/química , Enfermedades de las Plantas/microbiología , Unión Proteica , Propiedades de Superficie , Ulmus/microbiologíaAsunto(s)
Aterectomía Coronaria/efectos adversos , Aneurisma Coronario/diagnóstico , Hiperlipoproteinemia Tipo II/diagnóstico , Hiperlipoproteinemia Tipo II/patología , Anciano , Colesterol/sangre , Aneurisma Coronario/etiología , Aneurisma Coronario/terapia , Humanos , Masculino , Persona de Mediana EdadRESUMEN
In order to investigate the protein folding-unfolding process, dynamic light scattering (DLS) and atomic force microscopy (AFM) imaging were used to study two fragments of the muscle cardiac protein beta-connectin, also known as titin. Both fragments belong to the I band of the sarcomer, and they are composed of four domains from I(27) to I(30) (tetramer) and eight domains from I(27) to I(34) (octamer). DLS measurements provide the size of both fragments as a function of temperature from 20 up to 86 degrees C, and show a thermal denaturation due to temperature increase. AFM imaging of both fragments in the native state reveals a homogeneous and uniform distribution of comparable structures. The DLS and AFM techniques turn out to be complementary for size measurements of the fragments and fragment aggregates. An unexpected result is that the octamer folds into a smaller structure than the tetramer and the unfolded octamer is also smaller than the unfolded tetramer. This feature seems related to the significance of the hydrophobic interactions between domains of the fragment. The longer the fragment, the more easily the hydrophobic parts of the domains interact with each other. The fragment aggregation behavior, in particular conditions, is also revealed by both DLS and AFM as a process that is parallel to the folding-unfolding transition.
Asunto(s)
Modelos Químicos , Modelos Moleculares , Proteínas Musculares/química , Proteínas Musculares/ultraestructura , Miocardio/metabolismo , Fragmentos de Péptidos/química , Fragmentos de Péptidos/ultraestructura , Proteínas Quinasas/química , Proteínas Quinasas/ultraestructura , Simulación por Computador , Conectina , Humanos , Microscopía de Fuerza Atómica/métodos , Conformación Proteica , Refractometría/métodosRESUMEN
The stroma of human bone marrow contains a population of skeletal stem cells (hBM-MSC) which are common ancestors, among the others, of osteoblasts and adipocytes. It has been proposed that the imbalance between hBM-MSC osteogenesis and adipogenesis, which naturally accompanies bone marrow senescence, may contribute to the development of bone-associated diseases, like osteoporosis. The possibility to reproduce this mechanism in vitro has been demonstrated, providing a good model to disclose the details of the complex bone-fat generation homeostasis. Nevertheless, the lack of a simple approach to quantitatively assess the actual stage of a cellular population hindered the adoption of this in vitro model. In this work, the direct differentiation of hBM-MSCs towards a single (osteo or adipo) lineage was characterized using quantitative biophysical and biological approaches, together with the parallel process of trans-differentiation from one lineage to the other. The results confirm that the original plasticity of hBM-MSCs is maintained along the initial stages of the differentiation, showing that in vitro conversion of pre-osteoblasts into adipocytes and, vice versa, of pre-adipocytes into osteoblasts is extremely efficient, comparable with the direct differentiation. Moreover, a method based on digital holography is proposed, providing a quantitative indication of the phenotype stage along differentiation.
Asunto(s)
Transdiferenciación Celular , Células Madre Mesenquimatosas/citología , Modelos Biológicos , Osteoporosis/fisiopatología , Adipocitos/citología , Adipocitos/metabolismo , Fenómenos Biofísicos , Células de la Médula Ósea/citología , Diferenciación Celular , Linaje de la Célula , Células Cultivadas , Proteína de Unión a los Ácidos Grasos 7/genética , Proteína de Unión a los Ácidos Grasos 7/metabolismo , Holografía , Humanos , Imagenología Tridimensional , Células Madre Mesenquimatosas/metabolismo , Microscopía de Contraste de Fase , Osteoblastos/citología , Osteoblastos/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Osteoporosis/metabolismo , ARN/aislamiento & purificación , ARN/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteínas Supresoras de Tumor/genética , Proteínas Supresoras de Tumor/metabolismoRESUMEN
Eosinophilic granulomatosis with polyangitis (EGPA) is an uncommon ANCA-associated systemic small-vessel necrotizing vasculitis. At times, EGPA presenting manifestations can be very different from the usually recognized disease patterns. We report a 52-year-old female patient with 3 years history of itching. During the time occurred a chronic skin lichenification on her legs and gradually developed a full-blown ANCA-MPO positive EGPA in combination with blood hypereosinophilia, eosinophilic vasculitis at skin biopsy, subclinical asthma and chronic rhinosinusitis.
Asunto(s)
Síndrome de Churg-Strauss/complicaciones , Granulomatosis con Poliangitis/complicaciones , Liquen Plano/etiología , Enfermedad Crónica , Síndrome de Churg-Strauss/diagnóstico , Femenino , Granulomatosis con Poliangitis/diagnóstico , Humanos , Persona de Mediana EdadRESUMEN
To evaluate the relationship between carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) gut colonization and mortality in diabetic patients with a foot infection (DFI) we performed a single-centre, retrospective, matched case-control study. In the study period, we identified 21 patients with DFI who had KPC-Kp gut colonization and 21 controls. The 90-day mortality rate was significantly higher in patients with colonized guts (47%) than the controls (4%) (p 0.013). A multivariate analysis demonstrated that gut colonization with KPC-Kp was the only independent predictor of mortality: odds ratio 13.33, 95% CI 1.90-272.80, p 0.024. In patients with DFI, KPC-Kp gut colonization appears to be an important risk factor for mortality.
Asunto(s)
Portador Sano/microbiología , Pie Diabético/mortalidad , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/enzimología , beta-Lactamasas/metabolismo , Anciano , Anciano de 80 o más Años , Portador Sano/epidemiología , Estudios de Casos y Controles , Femenino , Humanos , Infecciones por Klebsiella/epidemiología , Klebsiella pneumoniae/aislamiento & purificación , Masculino , Factores de Riesgo , Análisis de SupervivenciaRESUMEN
AIM: Lipoprotein apheresis (LA) is the elective therapy for homozygous and other forms of familial hypercholesterolemia (FH) and familial combined hypercholesterolemia (FCH), resistant/intolerant to lipid lowering drugs, and hyperlipoproteinemia(a) for which drugs are not available. To assess the effect of LA on the incidence of adverse cardiac or vascular events (ACVE) at the time period of pre-initiation of apheresis and during the LA treatment. METHODS: We collected data of 30 patients (mean age 62 ± 8 years, males 73%), with FH, or FCH and cardiovascular disease on maximally tolerated lipid lowering therapy and LA treatment (median 5 years, interquartile range 3-8 years). Associated hyperlipoproteinemia(a) was present in 16/30 subjects. The LA treatment was performed biweekly as clinically indicated by dextran-sulfate or heparin-induced LDL precipitation apheresis. The ACVE incidence, before and after treatment, was evaluated by statistical analyses. RESULTS: The ACVE incidence occurred before and after the LA treatment inception, were 86 and 15 events respectively. Notably, 6/15 of ACVE were secondary to stent restenosis and 7/15 follow-up events occurred during the first 5 years. The AVCE rates/year were 0.58 and 0.13 respectively (p < 0.001). CONCLUSIONS: Our data confirm long-term efficacy and positive impact of LA on morbidity in patients with FH and FCH and atherosclerotic disease at maximally tolerated lipid lowering therapy.
Asunto(s)
Eliminación de Componentes Sanguíneos/métodos , Enfermedades Cardiovasculares/prevención & control , Hiperlipoproteinemias/terapia , Hipolipemiantes/uso terapéutico , Lipoproteínas/sangre , Anciano , Biomarcadores/sangre , Eliminación de Componentes Sanguíneos/efectos adversos , Enfermedades Cardiovasculares/diagnóstico , Enfermedades Cardiovasculares/epidemiología , Sulfato de Dextran/uso terapéutico , Femenino , Heparina/uso terapéutico , Humanos , Hiperlipoproteinemias/sangre , Hiperlipoproteinemias/diagnóstico , Hiperlipoproteinemias/epidemiología , Hipolipemiantes/efectos adversos , Incidencia , Italia/epidemiología , Masculino , Persona de Mediana Edad , Factores de Riesgo , Factores de Tiempo , Resultado del TratamientoRESUMEN
BACKGROUND: This study compared long-term survival in pancreatic or periampullary cancer treated with Whipple pancreatoduodenectomy (PD) and pylorus-preserving pancreatoduodenectomy (PPPD). METHODS: Two hundred twenty-one patients with pancreatic head or periampullary cancer were treated. Prognostic variables included age, gender, type and period of operation, and tumor stage. In the ductal adenocarcinomas variables also included tumor and node status, type of lymphadenectomy, pathologic grade, and presence of microscopic residual tumor. The end point was death as a result of neoplastic recurrence. Survival curves were estimated by using the Kaplan-Meier method, and multifactorial analysis was also performed on the data from the ductal adenocarcinoma group. RESULTS: The mortality rate was 8.2% in the PD group versus 7.0% in the PPPD group. Morbidity rates were 34.4% for PD and 45.8% for PPPD. Five-year survival was 9.6% in the ductal adenocarcinoma and 63.8% in the periampullary carcinoma groups. Univariate analysis failed to show statistically significant differences in survival curves between the two treatments in either patient group. Correcting for multiple variables in the ductal adenocarcinoma group did not reveal any significant differences in survival rates between the two treatments. CONCLUSIONS: PPPD was as successful as classic PD in the treatment of ductal adenocarcinoma and periampullary cancer of the pancreas. Long-term survival was not influenced by the type of resection.
Asunto(s)
Adenocarcinoma/cirugía , Carcinoma Ductal de Mama/cirugía , Neoplasias Pancreáticas/cirugía , Pancreaticoduodenectomía/métodos , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia , Complicaciones Posoperatorias , Píloro , Estudios Retrospectivos , Análisis de SupervivenciaRESUMEN
Mesenchymal stem cells have attracted great interest in the field of tissue engineering and regenerative medicine because of their multipotentiality and relative ease of isolation from adult tissues. The medical application of this cellular system requires the inclusion in a growth and delivery scaffold that is crucial for the clinical effectiveness of the therapy. In particular, the ideal scaffolding material should have the needed porosity and mechanical strength to allow a good integration with the surrounding tissues, but it should also assure high biocompatibility and full resorbability. For such a purpose, protein-inspired biomaterials and, in particular, elastomeric-derived polypeptides are playing a major role, in which they are expected to fulfil many of the biological and mechanical requirements. A specific chimeric protein, designed starting from elastin, resilin and collagen sequences, was characterized over different length scales. Single-molecule mechanics, aggregation properties and compatibility with human mesenchymal stem cells were tested, showing that the engineered compound is a good candidate as a stem cell scaffold to be used in tissue engineering applications.
Asunto(s)
Proteínas de la Matriz Extracelular/química , Proteínas de la Matriz Extracelular/metabolismo , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/fisiología , Péptidos/síntesis química , Péptidos/metabolismo , Andamios del Tejido , Proliferación Celular , Supervivencia Celular/fisiología , Células Cultivadas , Proteínas de la Matriz Extracelular/genética , Humanos , Péptidos/genética , Ingeniería de Proteínas/métodosRESUMEN
The three-dimensional structure and the mechanical properties of a ß-connectin fragment from human cardiac muscle, belonging to the I band, from I(27) to I(34), were investigated by small-angle x-ray scattering (SAXS) and single-molecule force spectroscopy (SMFS). This molecule presents an entropic elasticity behavior, associated to globular domain unfolding, that has been widely studied in the last 10 years. In addition, atomic force microscopy based SMFS experiments suggest that this molecule has an additional elastic regime, for low forces, probably associated to tertiary structure remodeling. From a structural point of view, this behavior is a mark of the fact that the eight domains in the I(27)-I(34) fragment are not independent and they organize in solution, assuming a well-defined three-dimensional structure. This hypothesis has been confirmed by SAXS scattering, both on a diluted and a concentrated sample. Two different models were used to fit the SAXS curves: one assuming a globular shape and one corresponding to an elongated conformation, both coupled with a Coulomb repulsion potential to take into account the protein-protein interaction. Due to the predominance of the structure factor, the effective shape of the protein in solution could not be clearly disclosed. By performing SMFS by atomic force microscopy, mechanical unfolding properties were investigated. Typical sawtooth profiles were obtained and the rupture force of each unfolding domain was estimated. By fitting a wormlike chain model to each peak of the sawtooth profile, the entropic elasticity of octamer was described.
Asunto(s)
Microscopía de Fuerza Atómica , Proteínas Musculares/química , Proteínas Quinasas/química , Dispersión del Ángulo Pequeño , Difracción de Rayos X , Conectina , Elasticidad , Humanos , Multimerización de Proteína , Estructura Cuaternaria de Proteína , Desplegamiento ProteicoAsunto(s)
Accidentes de Trabajo , Lesiones Encefálicas/terapia , Lóbulo Temporal/lesiones , Adulto , Lesiones Encefálicas/diagnóstico , Cuerpos Extraños/cirugía , Humanos , Masculino , Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/microbiologíaAsunto(s)
Proteínas Bacterianas/metabolismo , Infecciones por Klebsiella/epidemiología , Infecciones por Klebsiella/terapia , Klebsiella pneumoniae/enzimología , beta-Lactamasas/metabolismo , Anciano , Humanos , Unidades de Cuidados Intensivos , Masculino , Estudios Retrospectivos , Centros de Atención Terciaria , Resultado del TratamientoAsunto(s)
Catéteres/efectos adversos , Falla de Equipo , Embolia Pulmonar/etiología , Dispositivos de Acceso Vascular/efectos adversos , Anciano , Remoción de Dispositivos , Femenino , Humanos , Embolia Pulmonar/diagnóstico por imagen , Embolia Pulmonar/terapia , Radiografía , Resultado del TratamientoRESUMEN
Cerato-platanin (CP), the first member of the "cerato-platanin family", is a moderately hydrophobic protein produced by Ceratocystis fimbriata, the causal agent of a severe plant disease called "canker stain". The protein is localized in the cell wall of the fungus and it seems to be involved in the host-plane interaction and induces both cell necrosis and phytoalexin synthesis (one of the first plant defence-related events). Recently, it has been determined that CP, like other fungal surface protein, is able to self assemble in vitro. In this paper we characterize the aggregates of CP by Atomic Force Microscopy (AFM) images. We observe that CP tends to form early annular-shaped oligomers that seem to constitute the fundamental bricks of a hierarchical aggregation process, eventually resulting in large macrofibrillar assemblies. A simple model, based on the hypothesis that the aggregation is energetically favourable when the exposed surface is reduced, is compatible with the measured aggregates' shape and size. The proposed model can help to understand the mechanism by which CP and many other fungal surface proteins exert their effects.