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1.
Environ Sci Pollut Res Int ; 27(13): 14711-14722, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32052329

RESUMEN

Despite the exponential increase of studies on plastic debris in recent years, there are still few works focusing on the problem as it relates to inland waters: little is known about the accumulation and dispersion dynamics on lake shores, and there are no standardized sampling methods for monitoring purposes. The accumulation of plastic litter in natural habitats also threatens the resident organisms. In this paper, we investigated the abundance and accumulation of plastic particles, ranging in size from 1 to 50 mm, from the beach sediment of three volcanic lakes in Central Italy: Albano, Bracciano, and Vico. The collection was designed to define the most important variables that one must consider in order to obtain a representative sample of plastic litter in a lake environment. In view of the high heterogeneity of sampling protocols used, comparison among the obtained results is limited and sometimes impossible. By using one of the proposed sampling methodologies, and critically analyzing the results, we aimed to highlight a possible monitoring criterion and to identify specific elements that can be meaningful and representative. The samples were collected in May and September 2017. For each lake, we sampled plastic items and sediments from two beaches. Albano contained the largest amount of plastic (in weight), while Bracciano had the largest number of particles. Our observations lead us to infer that the number of particles is the parameter most sensitive to environmental variations, as well as the more suitable for monitoring with greater definition the differences between sites. Moreover, sampling should be taken in different seasons, following a sampling pattern that includes at least two beaches placed in strategic positions with respect to wind and waves.In order to identify new indicators to evaluate the entry points of plastic into the food web, we collected, from the same sites analyzed, some specimens of the Talitrid Amphipod Cryptorchestia garbinii, a detritivorous species having a critical role in debris turnover of these environments. To investigate the microplastic (MP) ingestion in natural conditions, we analyzed their digestive tracts with both Nile red staining method and micro-FTIR spectroscopy. The analyses confirmed that C. garbinii was able to ingest plastics in natural conditions. Therefore, it can signify one of the entry points for microplastics (MPs) in the trophic chain. This observation constitutes the first evidence of MP ingestion in this species.


Asunto(s)
Anfípodos , Contaminantes Químicos del Agua/análisis , Animales , Monitoreo del Ambiente , Italia , Lagos , Plásticos , Estaciones del Año
2.
Biomolecules ; 9(9)2019 09 17.
Artículo en Inglés | MEDLINE | ID: mdl-31533362

RESUMEN

Studying transcription machinery assembly in vitro is challenging because of long intrinsically disordered regions present within the multi-modular transcription factors. One example is alcohol dehydrogenase repressor 1 (Adr1p) from fermenting yeast, responsible for the metabolic switch from glucose to ethanol. The role of each individual transcription activation domain (TAD) has been previously studied, but their interplay and their roles in enhancing the stability of the protein is not known. In this work, we designed five unique miniAdr1 constructs containing either TADs I-II-III or TAD I and III, connected by linkers of different sizes and compositions. We demonstrated that miniAdr1-BL, containing only PAR-TAD I+III with a basic linker (BL), binds the cognate DNA sequence, located in the promoter of the ADH2 (alcohol dehydrogenase 2) gene, and is necessary to stabilize the heterologous expression. In fact, we found that the sequence of the linker between TAD I and III affected the solubility of free miniAdr1 proteins, as well as the stability of their complexes with DNA. miniAdr1-BL is the stable unit able to recognize ADH2in vitro, and hence it is a promising tool for future studies on nucleosomal DNA binding and transcription machinery assembly in vitro.


Asunto(s)
Alcohol Deshidrogenasa/genética , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/metabolismo , Ingeniería de Proteínas/métodos , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/enzimología , Factores de Transcripción/química , Factores de Transcripción/metabolismo , Alcohol Deshidrogenasa/química , Alcohol Deshidrogenasa/metabolismo , Sitios de Unión , Clonación Molecular , ADN de Hongos/metabolismo , Proteínas de Unión al ADN/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Pichia/genética , Pichia/metabolismo , Unión Proteica , Dominios Proteicos , Estabilidad Proteica , Saccharomyces cerevisiae/genética , Factores de Transcripción/genética , Activación Transcripcional
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