RESUMEN
Amyloid formation due to protein aggregation is associated with several amyloid diseases (amyloidosis). The use of small organic ligands as inhibitors of protein aggregation is an attractive strategy for the treatment of these diseases. In the present study, we evaluated the in vitro inhibitory and destabilizing effects of Mesalazine on human insulin fibrillation. To induce fibrillation, human insulin was incubated in 50 mM glycine buffer (pH 2.0) at 50 °C. The effect of Mesalazine on insulin amyloid aggregation was studied using spectroscopic, imaging, and computational approaches. Based on the results, the Mesalazine in a concentration-dependent manner (different ratios (1:0.1, 1:0.5, 1:1, and 1:5) of the insulin to Mesalazine) prevented the formation of amyloid fibrils and destabilized pre-formed fibrils. In addition, our molecular docking study confirmed the binding of Mesalazine to insulin through hydrogen bonds and hydrophobic interactions. Our findings suggest that Mesalazine may have therapeutic potential in the prevention of insulin amyloidosis and localized amyloidosis.
Asunto(s)
Amiloide , Amiloidosis , Humanos , Amiloide/química , Amiloide/metabolismo , Mesalamina/farmacología , Agregado de Proteínas , Simulación del Acoplamiento Molecular , Proteínas Amiloidogénicas , Insulina/metabolismo , Amiloidosis/tratamiento farmacológicoRESUMEN
BACKGROUND: Glyphosate is a non-selective systemic herbicide with a broad spectrum of weed control that inhibits a key enzyme, 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase, in the shikimate pathway. OBJECTIVES: Isolation and analysis of the epsps (aroA) gene responsible for glyphosate-tolerance in bacteria from Roundup- contaminated soils was the aim of this study. MATERIALS AND METHODS: Sampling was done from the soil of the gardens which were heavily contaminated by Roundup herbicide and then bacterial screening was performed in the presence of high concentrations of glyphosate. The genus of bacterium was identified via molecular methods such as 16S rRNA sequencing. The aroA gene of this bacterium (aroA HA-09) was isolated using the primers designed-upon specific regions of aroA genes available in NCBI GenBank database. The PCR product was cloned, sequenced and subcloned into pET28a as an expression vector and transferred into E. coli strain BL21(DE3). The cells were inoculated in liquid M9 minimal medium containing IPTG and different concentrations of glyphosate. RESULTS: The genus of bacterium was identified as Pseudomonas sp. strain HA-09. The isolated aroA HA-09 gene from this bacterium was approximately 2.2 kb in size. Bioassay of E. coli expressing this gene showed high tolerance to glyphosate (up to 300 mM). CONCLUSION: The aroA HA-09 gene could be considered as a novel and efficient candidate for development of glyphosate-tolerant crop plants.
RESUMEN
AIM: Squamous cell carcinoma of esophagus (SCCE) is an aggressive disease with a poor prognosis. Tropomyosins attach to actin microfilaments, providing its stability. Nonmuscle cells express Tpm isoforms such as Tpm1.6 and Tpm1.7 which are involved in cytoskeleton functional properties regulation. MATERIALS & METHODS: The expression of Tpm1.6 and Tpm1.7 was analyzed in SCCE tissues and its association with clinicopathological parameters and survival of patients was assessed. RESULTS: Tpm1.6 and Tpm1.7, besides TPM1 mRNA decreased considerably in SCCE tissues relative to normal esophageal tissues (p < 0.001). TPM1 downregulation level was significantly associated with the degree of tumor differentiation (p = 0.017). CONCLUSION: Tpm1.6 and Tpm1.7 suppression play a crucial role in esophagus tumorigenesis and could be associated with SCCE poor prognosis.
Asunto(s)
Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Tropomiosina/genética , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/genética , Regulación hacia Abajo , Esófago , Femenino , Humanos , Masculino , Persona de Mediana Edad , Isoformas de Proteínas/genética , ARN Mensajero/genética , Análisis de Supervivencia , Transcriptoma/genética , Tropomiosina/metabolismoRESUMEN
Thirty bacterial strains with various abilities to utilize glyphosate as the sole phosphorus source were isolated from farm soils using the glyphosate enrichment cultivation technique. Among them, a strain showing a remarkable glyphosate-degrading activity was identified by biochemical features and 16S rRNA sequence analysis as Ochrobactrum sp. (GDOS). Herbicide (3 mM) degradation was induced by phosphate starvation, and was completed within 60 h. Aminomethylphosphonic acid was detected in the exhausted medium, suggesting glyphosate oxidoreductase as the enzyme responsible for herbicide breakdown. As it grew even in the presence of glyphosate concentrations as high as 200 mM, Ochrobactrum sp. could be used for bioremediation purposes and treatment of heavily contaminated soils.
Asunto(s)
Glicina/análogos & derivados , Herbicidas/metabolismo , Ochrobactrum/aislamiento & purificación , Ochrobactrum/metabolismo , Microbiología del Suelo , Biodegradación Ambiental , ADN Bacteriano/genética , ADN Ribosómico/genética , Glicina/metabolismo , Datos de Secuencia Molecular , Ochrobactrum/clasificación , Ochrobactrum/genética , Filogenia , ARN Ribosómico 16S/genética , GlifosatoRESUMEN
Background: For successful in vitro plant regeneration, plant cell lines with multiple transgene integration and low transgene expression levels need to be ruled out. Although real-time polymerase chain reaction (RT-PCR) is a rapid way to accomplish this, it is also expensive and typically limits the size of the target sequence. Quantitative competitive PCR (QC-PCR) is proven to be a safe and accurate method for determination of both copy number and quantification of transcript levels of synthetic transgenes in transformed plants. Results: The glyphosate oxidoreductase genewas chemically synthesized and used to transform Brassica napus L. via Agrobactrium-mediated transformation. A construct containing the mutated form of a synthetic glyphosate oxidoreductase (gox) gene (internal standard) was prepared. Gene copy number was estimated in nine independent transgenic lines using QC-PCR as well as the standard method of Southern blot analysis. By quantitative RT-PCR, transcript levels were also determined in these lines. High (> 3), medium to high (2.2-3), medium to low (1-2.2), and low (< 1) levels of transcript were detected. Conclusions: No direct relationship was found between copy number and transgene expression levels. QC-PCR method could be implemented to screen putative transgenic plants and quickly select single T-DNA inserts. QC-PCR methods and the prepared competitor construct may be useful for future quantification of commercial transgenic food and feed.