RESUMEN
In this study, we investigated the effects of pingyangmycin (PYM) on the growth inhibition and apoptosis of human umbilical vein endothelial cells (HUVEC). In this study, we aimed to explore the optimal concentration of PYM to induce the apoptosis of HUVEC and to determine its mechanism of action. After treatment of HUVEC with different concentrations of PYM for 24 h, cell counting kit-8 (CCK-8) was used to detect growth inhibiting effects. Annexin V-FITC/propidium iodide stain was used to detect apoptosis, and western blot was used to detect the expression of glucose-related protein 78 (GPR78) and C/EBP homologous protein (CHOP) endoplasmic reticulum stress proteins. With increasing PYM concentration, the growth inhibition of HUVEC increased (P < 0.05), the apoptotic numbers of HUVEC increased (P < 0.05), with higher PYM concentrations inducing necrosis, and the protein expression of GRP78 and CHOP increased (P < 0.05). PYM could obviously inhibit the proliferation and promote the apoptosis of HUVEC. Necrotic cells were more prevalent than apoptotic cells at high PYM concentrations. This study helped to determine the proper concentration of PYM to induce more apoptosis than necrosis, which is critical to minimize inflammation, enhance the healing of the skin, and maintain safety for the patient. PYM might induce HUVEC apoptosis through the endoplasmic reticulum stress pathway.
Asunto(s)
Antibióticos Antineoplásicos/farmacología , Apoptosis , Bleomicina/análogos & derivados , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Bleomicina/farmacología , Línea Celular , Chaperón BiP del Retículo Endoplásmico , Estrés del Retículo Endoplásmico , Proteínas de Choque Térmico , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Humanos , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Factor de Transcripción CHOP/genética , Factor de Transcripción CHOP/metabolismoRESUMEN
An antifungal protein exhibiting a high activity against Sclerotinia sclerotiorum in vivo was purified by ammonium sulfate precipitation, hydrophobic chromatography, and gel filtration chromatography from the culture filtrate of the endophytic Bacillus subtilis strain Em7. The protein was characterized as a ß-1,3-1,4-glucanase according to amino acid analysis, and showed excellent properties in thermal stability and acid resistance. At the same time, the antifungal protein was cloned and heterologously expressed in Escherichia coli BL21. The recombinant protein was purified and showed similar enzymatic properties to the native protein, exhibiting strong inhibitory activity against S. sclerotiorum. This shows that the ß-1,3-1,4-glucanase may play a very important role in B. subtilis Em7 biocontrol function. In addition, many physiochemical properties of the native and purified recombinant protein were compared, including the effect of pH, temperature, metal cations, substrate specificity, and kinetic parameters. All parameters were similar between the native and recombinant purified protein, indicating that the purified recombinant protein has potential for industrial applications.
Asunto(s)
Antifúngicos/farmacología , Ascomicetos/efectos de los fármacos , Bacillus subtilis/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/farmacología , Proteínas Recombinantes , Proteínas Bacterianas/química , Proteínas Bacterianas/aislamiento & purificación , Activación Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Cinética , Pruebas de Sensibilidad Microbiana , Estabilidad Proteica , Especificidad por SustratoRESUMEN
The objectives of this study were to evaluate protein thermal stability, water-protein interaction, microstructure, and protein conformation between PSE-like and normal chicken breast meat batters. Sixty pale, soft, and exudative (PSE)-like (L*>53, pH24 h<5.7) and 60 normal (46Asunto(s)
Carne/análisis
, Músculos Pectorales/fisiología
, Conformación Proteica
, Animales
, Pollos/fisiología
, Electroforesis en Gel de Poliacrilamida
, Manipulación de Alimentos
, Calor
, Desnaturalización Proteica
RESUMEN
MicroRNAs (miRNAs) play critical roles in post-transcriptional gene regulation and act as important endogenous regulators to various stressors. Ultraviolet-B (UV-B) radiation is a major factor influencing crop growth and development. In this study, we isolated a novel wheat miRNA, named Tae-miR6000, and confirmed its expression diversity after UV-B treatments. Additionally, using the Northern blotting technique, we found that six miRNAs were highly responsive to UV-B stress in wheat. Of these six miRNAs, miR159, miR167a, and miR171 were significantly upregulated, and the remaining three miRNAs were downregulated, at different time points after UV-B treatment. This result indicates that miRNAs may be involved in the regulation of targets after induction by UV-B stress. Furthermore, promoter analysis of the UV-B-responsive miRNA genes revealed some light-relevant cis-elements, such as the I-box and G-box. Taken together, the results of this study suggest that wheat miRNAs play important roles in the response to UV-B stress.