Polyethylene terephthalate nanoparticles effect on RAW 264.7 macrophage cells

AbstractPlastic pollution is a major environmental concern due to its pervasiveness which continues to increase year on year, as a result of a continuing acceleration in global plastic production and use. Polyethylene terephthalate (PET) is among the most produced plastics, commonly used as food and beverage containers. Once released in the environment, the degradation of plastic materials produces micro-and nano-plastics, with a particular concern about potential toxicological effects if they cross epithelial barriers via inhalation or ingestion. In this work, the effect of PET nanoparticles (PET-NPs) (≤ 250 d.nm) was assayed on mouse macrophages cell line (RAW 264.7) in in vitro experiments. Results showed that PET nanoparticles were easily internalized by the cells, 15 μg/mL of nanoparticles concentration had exhibited effects in cell proliferation and a slightly increased production of Reactive Oxygen Species (ROS), which seems to trigger cell response as foreign particles related to upregulation of PCDH12, IGH-V10, ROBO1 genes, and cell maintenance functions, related to FTSJ2 gene upregulation. Thus, the RAW 264.7 results showed here are useful towards for a preliminary and understanding of the potentially toxic effects related to PET nanoparticles and complementary to other in vitro assays, as the first step into the development of the risk assessment framework

Characterization of an Alkali- and Halide-Resistant Laccase Expressed in E. coli: CotA from <i>Bacillus clausii</i>

The limitations of fungal laccases at higher pH and salt concentrations have intensified the search for new extremophilic bacterial laccases. We report the cloning, expression, and characterization of the bacterial cotA from Bacillus clausii, a supposed alkalophilic ortholog of cotA from B. subtilis. Both laccases were expressed in E. coli strain BL21(DE3) and characterized fully in parallel for strict benchmarking. We report activity on ABTS, SGZ, DMP, caffeic acid, promazine, phenyl hydrazine, tannic acid, and bilirubin at variable pH. Whereas ABTS, promazine, and phenyl hydrazine activities vs. pH were similar, the activity of B. clausii cotA was shifted upwards by ~0.5-2 pH units for the simple phenolic substrates DMP, SGZ, and caffeic acid. This shift is not due to substrate affinity (K(M)) but to pH dependence of catalytic turnover: The k(cat) of B. clausii cotA was 1 s⁻¹ at pH 6 and 5 s⁻¹ at pH 8 in contrast to 6 s⁻¹ at pH 6 and 2 s⁻¹ at pH 8 for of B. subtilis cotA. Overall, k(cat)/K(M) was 10-fold higher for B. subtilis cotA at pH(opt). While both proteins were heat activated, activation increased with pH and was larger in cotA from B. clausii. NaCl inhibited activity at acidic pH, but not up to 500-700 mM NaCl in alkaline pH, a further advantage of the alkali regime in laccase applications. The B. clausii cotA had ~20 minutes half-life at 80°C, less than the ~50 minutes at 80°C for cotA from B. subtilis. While cotA from B. subtilis had optimal stability at pH~8, the cotA from B. clausii displayed higher combined salt- and alkali-resistance. This resistance is possibly caused by two substitutions (S427Q and V110E) that could repel anions to reduce anion-copper interactions at the expense of catalytic proficiency, a trade-off of potential relevance to laccase optimization

Nanotecnología en procesos ambientales y remediación de la contaminación

El desarrollo de la nanotecnología ha abierto muchas oportunidades para la transformación de los procesos productivos. Sabiendo que los grandes retos de la humanidad son el de tener procesos productivos amigables al ambiente y energéticamente económicos, es de esperarse que la introducción de la nanotecnología contribuirá al cumplimiento de estas dos metas. Sin embargo, en la actualidad la sociedad sigue contaminando el ambiente poniendo en riesgo los diferentes ecosistemas y su misma salud. Existe la imperiosa necesidad de reducir, ver eliminar, el daño al ambiente y de limpiar y restaurar los sitios ya contaminados. En esta impostergable tarea, la nanotecnología tiene aún algo que decir y sobre todo hacer

Growth and production of cell constituents in batch cultures of <i>Botryococcus sudeticus</i>

The growth of, and the production of neutral lipids, carbohydrates and proteins by, the alga Botryococcus sudeticus in batch culture is described. The algal mass contains, during the stationary phase of growth, about 4.5% protein, 7.5% carbohydrate and 22.0% neutral lipid on a dry weight basis. Some physiological characteristics of this species are discussed

Microarray analysis of Neosartorya fischeri using different carbon sources, petroleum asphaltenes and glucose-peptone

Asphaltenes are considered as the most recalcitrant petroleum fraction and represent a big problem for the recovery, separation and processing of heavy oils and bitumens. Neosartorya fischeri is a saprophytic fungus that is able to grow using asphaltenes as the sole carbon source [1]. We performed transcription profiling using a custom designed microarray with the complete genome from N. fischeri NRRL 181 in order to identify genes related to the transformation of asphaltenes [1]. Data analysis was performed using the genArise software. Results showed that 287 genes were up-regulated and 118 were down-regulated. Here we describe experimental procedures and methods about our dataset (NCBI GEO accession number GSE68146) and describe the data analysis to identify different expression levels in N. fischeri using this recalcitrant carbon source