Comparison of methods for measuring shear force and sarcomere length and their relationship with sensorial tenderness of longissimus muscle in beef.

Slice shear force (SSF) and laser diffraction, considered faster methodologies, for measuring beef instrumental tenderness and sarcomere length, were compared with reference methodologies Warner-Bratzler shear force (WBSF) and phase contrast microscopy. Striploin samples (n = 74) were analyzed for pH, sarcomere length, instrumental tenderness, myofibrillar fragmentation index, and sensorial tenderness. Pearson's correlation measured the association of meat evaluation methods with residual analysis of the multivariate analysis of variance model. The n-dimensional profile to evaluated methods was presented by biplot to identify the behavior of the correlation between the methods (variables). There was moderate correlation between SSF and WBSF (r = .63; p < .01) and both presented moderate correlation with sensorial tenderness (r = - .62 and -.55, to SSF and WBSF, respectively; p < .01). However, WBSF was more efficient to classify samples as tender (68%) than SSF (47%), comparing with sensorial tenderness (80%). There was a moderate correlation for laser and microscopy for sarcomere length (r = .57; p < .01). Sarcomeres were shorter when measured by laser than microscopy. Either with low correlation coefficients, sarcomere measured by laser (r = .29; p < .05) presented higher correlation with sensorial tenderness than with microscopy (r = .22; p < .10). Results highlighted that SSF was faster and easier to run, while WBSF was more appropriate to classify samples by sensorial tenderness grades. Laser diffraction is more suitable to explain effects on tenderness; however, microscopy revealed results of sarcomere length that were more realistic, once laser can underestimate sarcomeres.

In vivo measurements reveal a single 5'-intron is sufficient to increase protein expression level in Caenorhabditis elegans.

Introns can increase gene expression levels using a variety of mechanisms collectively referred to as Intron Mediated Enhancement (IME). IME has been measured in cell culture and plant models by quantifying expression of intronless and intron-bearing reporter genes in vitro. We developed hardware and software to implement microfluidic chip-based gene expression quantification in vivo. We altered position, number and sequence of introns in reporter genes controlled by the hsp-90 promoter. Consistent with plant and mammalian studies, we determined a single, natural or synthetic, 5'-intron is sufficient for the full IME effect conferred by three synthetic introns, while a 3'-intron is not. We found coding sequence can affect IME; the same three synthetic introns that increase mcherry protein concentration by approximately 50%, increase mEGFP by 80%. We determined IME effect size is not greatly affected by the stronger vit-2 promoter. Our microfluidic imaging approach should facilitate screens for factors affecting IME and other intron-dependent processes.