Saturated fatty acids contribute to beta-cell dysfunction in the onset of type 2 diabetes mellitus. Cellular responses to lipotoxicity include oxidative stress, endoplasmic reticulum (ER) stress, and blockage of autophagy. Palmitate induces ER Ca2+ depletion followed by notable store-operated Ca2+ entry. Subsequent elevation of cytosolic Ca2+ can activate undesirable signaling pathways culminating in cell death. Mitochondrial Ca2+ uniporter (MCU) is the major route for Ca2+ uptake into the matrix and couples metabolism with insulin secretion. However, it has been unclear whether mitochondrial Ca2+ uptake plays a protective role or contributes to lipotoxicity. Here, we observed palmitate upregulated MCU protein expression in a mouse clonal beta-cell, MIN6, under normal glucose, but not high glucose medium. Palmitate elevated baseline cytosolic Ca2+ concentration ([Ca2+],) and reduced depolarization-triggered Ca2+ influx likely due to the inactivation of voltage-gated Ca2+ channels (VGCCs). Targeted reduction of MCU expression using RNA interference abolished mitochondrial superoxide production but exacerbated palmitate-induced [Ca2+], overload. Consequently, MCU knockdown aggravated blockage of autophagic degradation. In contrast, co-treatment with verapamil, a VGCC inhibitor, prevented palmitate-induced basal [Ca2+], elevation and defective [Ca2+], transients. Extracellular Ca2+ chelation as well as VGCC inhibitors effectively rescued autophagy defects and cytotoxicity. These observations suggest enhanced mitochondrial Ca2+ uptake via MCU upregulation is a mechanism by which pancreatic beta-cells are able to alleviate cytosolic Ca2+ overload and its detrimental consequences.

Inorganic phosphate (Pi) is essential for maintaining cellular function but excess of Pi leads to serious complications, including vascular calcification. Accumulating evidence suggests that oxidative stress contributes to the pathogenic progression of calcific changes. However, the molecular mechanism underlying Pi-induced reactive oxygen species (ROS) generation and its detrimental consequences remain unclear. Type III Na⁺-dependent Pi cotransporter, PiT-1/-2, play a significant role in Pi uptake of vascular smooth muscle cells. Pi influx via PiT-1/-2 increases the abundance of PiT-1/-2 and depolarization-activated Ca²⁺ entry due to its electrogenic properties, which may lead to Ca²⁺ and Pi overload and oxidative stress. At least four mitochondrial Pi transporters are suggested, among which the phosphate carrier (PiC) is known to be mainly involved in mitochondrial Pi uptake. Pi transport via PiC may induce hyperpolarization and superoxide generation, which may lead to mitochondrial dysfunction and endoplasmic reticulum stress, together with generation of cytosolic ROS. Increase in net influx of Ca²⁺ and Pi and their accumulation in the cytosol and mitochondrial matrix synergistically increases oxidative stress and osteogenic differentiation, which could be prevented by suppressing either Ca²⁺ or Pi overload. Therapeutic strategies targeting plasmalemmal and mitochondrial Pi transports can protect against Pi-induced oxidative stress and vascular calcification.

Heart failure (HF) is a global epidemic with a rising burden on individuals and healthcare systems. Advanced HF is characterized by severe symptoms at rest and marked limitations in physical activity. Stem cell therapies potentially address the limitations of current conventional treatments by reversing damage to cardiac tissue via their unique capacity for self-renewal and multilineage differentiation.

Single-incision laparoscopic endorectal pull-through (SILEP) has emerged as an alternative to conventional laparoscopic surgery for Hirschsprung disease (HSCR), offering improved cosmetic outcomes with comparable efficacy. However, its adoption remains limited, and evidence supporting its long-term safety and functional outcomes is sparse. This study presents our institutional experience with SILEP, alongside a systematic review and meta-analysis evaluating perioperative outcomes, postoperative complications, bowel function, and cosmesis. A prospective study was conducted at our institution involving 93 pediatric patients diagnosed with HSCR, who underwent SILEP between 2020 and 2021. Demographic data, perioperative details, and postoperative outcomes, including bowel function score (Rintala) and cosmetic results (Manchester Scar Scale), were prospectively collected and analyzed. Additionally, a systematic review and meta-analysis were performed following PRISMA guidelines, pooling data from nine studies comprising 420 patients. The mean operative time was 70.0 ± 30.3 min, significantly shorter than the pooled estimate from the meta-analysis (166.44 min; 95% CI 137.74-195.14, p < 0.001). No intraoperative complications or conversions were recorded. The mean hospital stay was 5.23 ± 2.69 days, consistent with prior reports. Enterocolitis occurred in 16.1% of cases, comparable to the pooled estimate (12%; 95% CI 7-21%). Among patients aged ≥ 4 years (n = 64), 84% achieved good to excellent bowel function scores. The mean Manchester Scar Score was 5.73 ± 1.38. SILEP is a safe, effective, and cosmetically superior approach for HSCR, with favorable perioperative outcomes and functional results. Our findings, supported by meta-analysis data, reinforce the feasibility of SILEP as a viable alternative for HSCR management.

Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are a powerful tool in advancing cardiovascular research and clinical applications. The maturation of sarcomere organization in hiPSC-CMs is crucial, as it supports the contractile function and structural integrity of these cells. Traditional methods for assessing this maturation like manual annotation and feature extraction are labor-intensive, time-consuming, and unsuitable for high-throughput analysis. To address this, we propose D-SarcNet, a dual-stream deep learning framework that takes fluorescent hiPSC-CM single-cell images as input and outputs the stage of the sarcomere structural organization on a scale from 1.0 to 5.0. The framework also integrates Fast Fourier Transform (FFT), deep learning-generated local patterns, and gradient magnitude to capture detailed structural information at both global and local levels. Experiments on a publicly available dataset from the Allen Institute for Cell Science show that the proposed approach not only achieves a Spearman correlation of 0.868 marking a 3.7% improvement over the previous state-of-the-art but also significantly enhances other key performance metrics, including MSE, MAE, and R2 score. Beyond establishing a new state-of-the-art in sarcomere structure assessment from hiPSC-CM images, our ablation studies highlight the significance of integrating global and local information to enhance deep learning networks ability to discern and learn vital visual features of sarcomere structure.