Conditioning of hiPSC-derived cardiomyocytes using surface topography obtained with high throughput technology.

Surface functionalization of polymers aims to introduce novel properties that favor bioactive responses. We have investigated the possibility of surface functionalization of polyethylene terephthalate (PET) sheets by the combination of laser ablation with hot embossing and the application of such techniques in the field of stem cell research. We investigated the response of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to topography in the low micrometer range. HiPSC-CMs are expected to offer new therapeutic tools for myocardial replacement or regeneration after an infarct or other causes of cardiac tissue loss. However, hiPSC-CMs are phenotypically immature compared to myocytes in the adult myocardium, hampering their clinical application. We aimed to develop and test a high-throughput technique for surface structuring that would improve hiPSC-CMs structural maturation. We used laser ablation with a ps-laser source in combination with nanoimprint lithography to fabricate large areas of homogeneous micron- to submicron line-like pattern with a spatial period of 3 µm on the PET surface. We evaluated cell morphology, alignment, sarcomeric myofibrils assembly, and calcium transients to evaluate phenotypic changes associated with culturing hiPSC-CMs on functionalized PET. Surface functionalization through hot embossing was able to generate, at low cost, low micrometer features on the PET surface that influenced the hiPSC-CMs phenotype, suggesting improved structural and functional maturation. This technique may be relevant for high-throughput technologies that require conditioning of hiPSC-CMs and may be useful for the production of these cells for drug screening and disease modeling applications with lower costs.

Metabolic responses of channel catfish (Ictalurus punctatus) exposed to phenol and post-exposure recovery.

Metabolic adjustments were studied in channel catfish Ictalurus punctatus exposed to 1.5 mg L-1 of phe nol (10% LC50) for four days and recovered for seven days. Lower triacylglycerol (TGA) stores and increased muscle fat free acids (FFA) suggest fat catabolism in muscle. Remarkable liver FFA decrease (-31%) suggests liver fat catabolism as well. Increased muscular ammonia levels and ASAT (aspartate aminotransferase) and decreased plasma aminoacids suggest higher muscular amino acid uptake. Constant levels of glucose and increased liver glycogen stores, associated with lower amino acids in plasma, indicate gluconeogenesis from amino acids. This is supported by higher hepatic ALAT and ASAT. Higher hepatic LDH followed by lower plasma lactate may indicate that plasma lactate was also used as gluconeogenic substrate. Biochemical alterations were exacerbated during the post-exposure recovery period. Reduction in muscle and plasma protein content indicate proteolysis. A higher rate of liver fat catabolism was resulted from a remarkable decrease in hepatic TGA (-58%). Catabolic preference for lipids was observed in order to supply such elevated energy demand. This study is the first insight about the metabolic profile of I. punctatus to cope with phenol plus its ability to recover, bringing attention to the biological consequences of environmental contamination.

Metabolic effects of trichlorfon (Masoten®) on the neotropical freshwater fish pacu (Piaractus mesopotamicus).

Fish parasites are among the crucial limiting factors in aquaculture. The organophosphorous pesticide trichlorfon is widely used as an insecticide and against fish parasites worldwide. In this study, the effects of environmental trichlorfon on biochemical and physiological parameters were investigated in Piaractus mesopotamicus (pacu), a widely farmed fish in South America, through sublethal exposure (8 µg L(-1), 10 % of the LC50; 96 h) and recovery. The activity of brain acetylcholinesterase (AChE) was reduced after exposure (15.5 %) and remained decreased during the recovery (21.5 %). In white muscle, AChE activity decreased 31 % only after recovery. Alkaline phosphatase (ALP) and acid phosphatase (ACP) activities of the liver, muscle and plasma were steady during exposure. However, after the recovery period, ALP activity was increased in the liver and muscle and decreased in plasma, while ACP was increased in the liver and decreased in muscle. Intermediary metabolism was also affected by trichlorfon, depicting increase of energetic demand (hypoglycemia, neoglucogenesis and lipid catabolism), which remained even after recovery. These results indicate that P. mesopotamicus is adversely affected by sublethal concentrations of trichlorfon and are useful for assessing the impact as well as the pros and cons of its use in controlling fish parasites in aquaculture.