RESUMO
Sepsis, a life-threatening inflammatory response, demands economical, accurate, and rapid detection of biomarkers during the critical "golden hour" to reduce the patient mortality rate. Here, we demonstrate a cost-effective waveguide-enhanced nanogold-linked immunosorbent assay (WENLISA) based on nanoplasmonic waveguide biosensors for the rapid and sensitive detection of procalcitonin (PCT), a sepsis-related inflammatory biomarker. To enhance the limit of detection (LOD), we employed sandwich assays using immobilized capture antibodies and detection antibodies conjugated to gold nanoparticles to bind the target analyte, leading to a significant evanescent wave redistribution and strong nanoplasmonic absorption near the waveguide surface. Experimentally, we detected PCT for a wide linear response range of 0.1 pg/mL to 1 ng/mL with a record-low LOD of 48.7 fg/mL (3.74 fM) in 8 min. Furthermore, WENLISA has successfully identified PCT levels in the blood plasma of patients with sepsis and healthy individuals, offering a promising technology for early sepsis diagnosis.
Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Sepse , Humanos , Pró-Calcitonina , Imunoadsorventes , Ouro , Sepse/diagnóstico , Biomarcadores , Anticorpos ImobilizadosRESUMO
Effects of leukemia inhibitory factor (LIF) and fibroblast growth factor 2 (FGF2) on establishment and maintenance of rabbit embryonic stem cell (rESC) lines were assessed. When grown on MEF feeders, rESC lines derived from fertilized embryos were established and maintained in medium containing paracrine factors LIF (via STAT3) and/or FGF2 (via MEK-ERK1/2 and PI3K-AKT). However, high levels of ERK1/2 and AKT activities in rESCs were crucial for maintaining their undifferentiated proliferation. Although rESCs under the influence of either LIF (500, 1,000, and 2,000 U/ml) or FGF2 (5, 10, and 20 ng/ml) alone had enhanced expression of pluripotency markers, peak expression occurred when both LIF (1,000 U/ml) and FGF2 (10 ng/ml) were applied. Induced dephosphorylation of STAT3, ERK1/2, and AKT by specific inhibitors limited growth of rESCs and caused remarkable losses of self-renewal capacity; therefore, we inferred that STAT3, ERK, and AKT had essential roles in maintaining rESC proliferation and self-renewal. We concluded that LIF and FGF2 jointly maintained the undifferentiated state and self-renewal of rESCs through an integrative signaling module.
Assuntos
Proliferação de Células/efeitos dos fármacos , Células-Tronco Embrionárias/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/farmacologia , Fator Inibidor de Leucemia/farmacologia , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/metabolismo , Proteínas de Homeodomínio/metabolismo , Camundongos , Proteína Quinase 1 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fator 3 de Transcrição de Octâmero/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores de Fosfoinositídeo-3 Quinase , Fosforilação/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo , Coelhos , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Zigoto/citologiaRESUMO
The growth and on-site bioremediation potential of an isolated thermal- and CO2-tolerant mutant strain, Chlorella sp. MTF-7, were investigated. The Chlorella sp. MTF-7 cultures were directly aerated with the flue gas generated from coke oven of a steel plant. The biomass concentration, growth rate and lipid content of Chlorella sp. MTF-7 cultured in an outdoor 50-L photobioreactor for 6 days was 2.87 g L⻹ (with an initial culture biomass concentration of 0.75 g L⻹), 0.52 g L⻹ d⻹ and 25.2%, respectively. By the operation with intermittent flue gas aeration in a double-set photobioreactor system, average efficiency of CO2 removal from the flue gas could reach to 60%, and NO and SO2 removal efficiency was maintained at approximately 70% and 50%, respectively. Our results demonstrate that flue gas from coke oven could be directly introduced into Chlorella sp. MTF-7 cultures to potentially produce algal biomass and efficiently capture CO2, NO and SO2 from flue gas.
Assuntos
Poluentes Atmosféricos/metabolismo , Biomassa , Chlorella/crescimento & desenvolvimento , Microalgas/crescimento & desenvolvimento , Biodegradação Ambiental , Dióxido de Carbono/metabolismo , Chlorella/metabolismo , Metalurgia , Microalgas/metabolismo , Óxidos de Nitrogênio/metabolismo , Dióxido de Enxofre/metabolismoRESUMO
Diabetes mellitus (DM), a state of chronic hyperglycemia, is associated with a variety of serious complications. Hyperglycemia-induced advanced glycation end products (AGEs) play an important role in the development of diabetic complications. In vivo, we demonstrated that disrupted mitochondria and autophagy was elevated in type II DM db/db mice. Mitophagy was evidenced by increased autophagosome formation in the beta-islet cells. The adducts of N(epsilon)-(carboxymethyl) lysine (CML), a major AGE, and bovine serum albumin (CML-BSA) stimulated the conversion of microtubule-associated protein 1 light chain 3-I (LC3-I) to LC3-II in rat insulinoma cells (RIN-m5F). CML-BSA increased ROS generation as demonstrated in a time-dependent manner. Experiments with mitochondrial targeted enhanced yellow fluorescent protein transfected RIN-m5F cells, massive fragmented mitochondria were visualized in the CML-BSA treated cells. Taken together, these data suggested that AGEs may cause mitochondrial dysfunction and mitophagosome formation, and AGEs-induced glycoxidative stress may trigger mitophagic process to modulate mitochondrial fates leading to either cell survival or cell death.