Comparative study of ultrasound application versus mechanical agitation on pork belly brining for bacon production.

Pork belly brining is a time-consuming step of bacon production that needs to be studied and enhanced through suitable technologies. In this sense, this study aimed at evaluating the impact of ultrasound (US), mechanical agitation (AG), and static brine (SB) on the kinetics of water loss (WL), solids gain (SG), and salt content (SC) of pork belly during brining under different temperatures. Mathematical models were used to estimate mass transfer rates, equilibrium parameters, and thermodynamic properties. Peleg model was chosen as the most suitable model to predict the kinetics experimental data (Radj2 ≥ 0.979 and RMSE ≤ 0.014). The increase in the brine temperature increased WL, SG, and SC for all treatments. Nonlinear effects of temperature were observed for WL, SG, and SC, following an Arrhenius-type behavior. The assistance of ultrasound significantly enhanced the velocity of WL, SG, and SC by 32-56%, while AG improved by 18-39% both compared to SB. Brining was considered an endothermic and non-spontaneous process through the thermodynamic assessment. The increase in temperature and the AG and US processes accelerated the formation of the activated complex. The application of ultrasound was considered the most suitable technology to reduce the brining time. However, significant improvements can be obtained by mechanical agitation. Therefore, both methods can be used to reduce the time processing of pork belly aiming at accelerating the bacon production process.

Gene-Edited Human-Induced Pluripotent Stem Cell Lines to Elucidate DAND5 Function throughout Cardiac Differentiation.

(1) Background: The contribution of gene-specific variants for congenital heart disease, one of the most common congenital disabilities, is still far from our complete understanding. Here, we applied a disease model using human-induced pluripotent stem cells (hiPSCs) to evaluate the function of DAND5 on human cardiomyocyte (CM) differentiation and proliferation. (2) Methods: Taking advantage of our DAND5 patient-derived iPSC line, we used CRISPR-Cas9 gene-editing to generate a set of isogenic hiPSCs (DAND5-corrected and DAND5 full-mutant). The hiPSCs were differentiated into CMs, and RT-qPCR and immunofluorescence profiled the expression of cardiac markers. Cardiomyocyte proliferation was analysed by flow cytometry. Furthermore, we used a multi-electrode array (MEA) to study the functional electrophysiology of DAND5 hiPSC-CMs. (3) Results: The results indicated that hiPSC-CM proliferation is affected by DAND5 levels. Cardiomyocytes derived from a DAND5 full-mutant hiPSC line are more proliferative when compared with gene-corrected hiPSC-CMs. Moreover, parallel cardiac differentiations showed a differential cardiac gene expression profile, with upregulated cardiac progenitor markers in DAND5-KO hiPSC-CMs. Microelectrode array (MEA) measurements demonstrated that DAND5-KO hiPSC-CMs showed prolonged field potential duration and increased spontaneous beating rates. In addition, conduction velocity is reduced in the monolayers of hiPSC-CMs with full-mutant genotype. (4) Conclusions: The absence of DAND5 sustains the proliferation of hiPSC-CMs, which alters their electrophysiological maturation properties. These results using DAND5 hiPSC-CMs consolidate the findings of the in vitro and in vivo mouse models, now in a translational perspective. Altogether, the data will help elucidate the molecular mechanism underlying this human heart disease and potentiates new therapies for treating adult CHD.

Generation of a gene-corrected human induced pluripotent stem cell line derived from a patient with laterality defects and congenital heart anomalies with a c.455G > A alteration in DAND5.

Human induced pluripotent stem cells (hiPSCs) from individual patient basis are considered a powerful resource to model human diseases. However, to study complex multigenic diseases such as Congenital Heart Disease, it is crucial to generate perfect isogenic controls to understand gene singularity and contribution. Here, we report the engendering of an isogenic hiPSC line with homozygous correction of c.455G > A alteration in the DAND5 gene, using CRISPR/Cas9 technology. The characterization of a clone of this cell line demonstrates normal karyotype, pluripotent state, and potential to differentiate in vitro towards endoderm, mesoderm, and ectoderm.