We have shown that the miR(21/10b) ratio is an independent prognostic factor for M0 ccRCC patients, which could be useful to identify high-risk M0 patients who could benefit from increased surveillance.

Rat embryos at all cleavage stages of development were cultured in vitro. The various cleavage stages were successfully cultured through at least one mitotic cycle with the exception of the four-cell stage. No instance of development through the four-cell stage was observed. The eight-cell stage was cultured to the blastocyst stage with about 80 % success. There was no difference in viability of eight-cell embryos if they were cultured without the zona pellucida.Despite some difficulties with adhesion of embryos and cellular death, denuded embryo pairs aggregated into a single blastocyst during a 24-hr culture period. Of all such aggregation attempts, 45 % proved successful. Twenty-five aggregate blastocysts were transferred to eight recipient females. Subsequently, seven transferred embryos were born to four of the host females.Five of these transferred embryos were externally female while the remaining two were normal males. Two of these females were sibling aggregates (identical phenotypes) and no evidence of mosaicism should have been observed. Of the remaining five animals, three were of one parental phenotype or the other, while the remaining two (one male and one female) expressed some degree of parental phenotype mosaicism.

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Improvement of imaging quality has the potential to visualize previously unseen building blocks of the brain and is therefore one of the great challenges in neuroscience. Rapid development of new tissue clearing techniques in recent years have attempted to solve imaging compromises in thick brain samples, particularly for high resolution optical microscopy, where the clearing medium needs to match the high refractive index of the objective immersion medium. These problems are exacerbated in insect tissue, where numerous (initially air-filled) tracheal tubes branching throughout the brain increase the scattering of light. To date, surprisingly few studies have systematically quantified the benefits of such clearing methods using objective transparency and tissue shrinkage measurements. In this study we compare a traditional and widely used insect clearing medium, methyl salicylate combined with permanent mounting in Permount (“MS/P”) with several more recently applied clearing media that offer tunable refractive index (n): 2,2′-thiodiethanol (TDE), “SeeDB2” (in variants SeeDB2S and SeeDB2G matched to oil and glycerol immersion, n = 1.52 and 1.47, respectively) and Rapiclear (also with n = 1.52 and 1.47). We measured transparency and tissue shrinkage by comparing freshly dissected brains with cleared brains from dipteran flies, with or without addition of vacuum or ethanol pre-treatments (dehydration and rehydration) to evacuate air from the tracheal system. The results show that ethanol pre-treatment is very effective for improving transparency, regardless of the subsequent clearing medium, while vacuum treatment offers little measurable benefit. Ethanol pre-treated SeeDB2G and Rapiclear brains show much less shrinkage than using the traditional MS/P method. Furthermore, at lower refractive index, closer to that of glycerol immersion, these recently developed media offer outstanding transparency compared to TDE and MS/P. Rapiclear protocols were less laborious compared to SeeDB2, but both offer sufficient transparency and refractive index tunability to permit super-resolution imaging of local volumes in whole mount brains from large insects, and even light-sheet microscopy. Although long-term permanency of Rapiclear stored samples remains to be established, our samples still showed good preservation of fluorescence after storage for more than a year at room temperature.