Effects of caloric restriction on oxidative stress parameters

Takayasu Arteritis (TA) is characterized by granulomatous panarteritis, vessel wall fibrosis, and irreversible vascular impairment. The aim of this study is to explore the usefulness of the Enhanced Liver Fibrosis score (ELF), procollagen-III aminoterminal propeptide (PIIINP), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), and hyaluronic acid (HA) in assessing vascular damage in TA patients. ELF, PIIINP, TIMP-1, and HA were measured in 24 TA patients, and the results were correlated with the clinical damage indexes (VDI and TADS), an imaging damage score (CARDS), and disease activity scores (NIH and ITAS2010). A mean ELF score 8.42 (±1.12) and values higher than 7.7 (cut-off for liver fibrosis) in 21/24 (87.5%) of patients were detected. The VDI and TADS correlated significantly to ELF (p < 0.01). Additionally, a strong association across ELF and CARDS (p < 0.0001), PIIINP and CARDS (p < 0.001), and HA and CARDS (p < 0.001) was observed. No correlations of the tested biomarkers with inflammatory parameters, NIH, and ITAS2010 scores were found. To our knowledge, this is the first study that suggests the association of the serum biomarkers PIIINP, HA, and ELF score with damage but not with disease activity in TA patients. The ELF score and PIIINP may be useful biomarkers reflecting an ongoing fibrotic process and quantifying vascular damage.

Zebrafish (Danio rerio) are widely used as a vertebrate model in cardiovascular research due to their genetic similarity to humans, optical transparency during early development, and amenability to in vivo imaging. This manuscript presents a standardized, accessible protocol for assessing cardiac morphology and function in zebrafish embryos at 96 h post-fertilization (hpf) using brightfield light microscopy. The method includes embryo collection, morphological screening, immobilization in agarose, video recording of the beating heart, and image-based analysis of cardiac parameters such as ventricular dimensions, stroke volume, heart rate, ejection fraction, and cardiac output. Calculations are based on geometric approximations of the ventricle using open-source software (e.g., ImageJ, Zembryo Analyzer). The protocol enables quantitative assessment of cardiac performance using widely available equipment, making it suitable for laboratories with limited resources and high-throughput screenings. In this study, 96 hpf zebrafish larvae were analyzed by recording the beating heart under a light microscope. Key cardiac parameters measured included ventricular dimensions, which were used to calculate stroke volume and heart rate, determined by beats per minute. From these, ejection fraction and cardiac output were derived to assess overall cardiac performance. Representative results from healthy embryos demonstrated consistent ventricular contraction and robust functional indices, while embryos exposed to cadmium showed impaired cardiac morphology and significantly reduced cardiac output. Overall, the method is important for detecting developmental cardiotoxicity and provides a reproducible, non-invasive approach for evaluating cardiac function in vivo during early zebrafish development.