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All Journal Jurnal Widya Medika
Jaya, Ferdinand Wiliam
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Medicine & Pharmacology

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PROTOCOL OPTIMIZATION OF RT-qPCR FOR NF-ΚB DETECTION IN CARDIAC TISSUE FROM HFHS-FED MICE Jaya, Ferdinand Wiliam; Novita, Bernadette Dian; Suwandito, Laurensius; Wijaya, Hendy; Tjahjono, Yudy; Hendrata, Adi Pramono; Dewi, Sianty; Wijaya, Sumi; Ervina, Martha; Kuncorojakti, Suryo
JURNAL WIDYA MEDIKA Vol. 12 No. 1 (2026): March
Publisher : FAKULTAS KEDOKTERAN UNIVERSITAS KATOLIK WIDYA MANDALA SURABAYA

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33508/jwm.v12i1.8196

Abstract

Accurate detection of gene expression in challenging tissues requires methodological optimization to ensure reproducibility. Cardiac tissue, with its low cell density and variable RNA yield, presents particular difficulties for RT‑qPCR analysis. This study aimed to refine NF‑κB detection in HFHS‑fed C57BL/6 mice by comparing a standard 2‑ΔΔCt approach with an optimized protocol incorporating individualized calibrators adjusted for RNA concentration. Six male mice were divided into standard-diet (STD) and high‑fat, high‑sucrose (HFHS) groups, and cardiac RNA was extracted after 8 weeks of feeding. While relative NF‑κB expression trends were higher in the HFHS group, differences were not statistically significant; however, the optimized method consistently reduced variability and improved measurement reliability compared to the generalized approach. These findings highlight the value of individualized calibration as a protocol refinement for RT‑qPCR, offering a more rational framework for gene expression analysis in heterogeneous or technically demanding tissue samples.
IMMUNOMODULATORY EFFECTS OF HUMAN ALLOGENEIC MESENCHYMAL STEM CELLS IN AGING-RELATED FRAILTY: A SYSTEMATIC REVIEW AND META-ANALYSIS OF RANDOMIZED CONTROLLED TRIALS Wedharga, I Gede Putu Adhi; Jaya, Ferdinand Wiliam
JURNAL WIDYA MEDIKA Vol. 12 No. 2 (2026): July
Publisher : FAKULTAS KEDOKTERAN UNIVERSITAS KATOLIK WIDYA MANDALA SURABAYA

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33508/jwm.v12i2.8384

Abstract

Aging-related frailty is associated with immune dysregulation, including chronic inflammation and immunosenescence. Human allogeneic mesenchymal stem cells (hMSCs) have been proposed as immunomodulatory therapies; however, the responsive immune domains remain unclear. This systematic review and meta-analysis explored immune and inflammatory biomarkers reported in randomized controlled trials (RCTs) of hMSC therapy for aging-related frailty. Major databases were searched through April 2026 for RCTs enrolling adults aged ≥60 years with clinically diagnosed frailty. Outcomes encompassed innate inflammation, adaptive immune activation and composition, immunosenescence-related measures, and alloimmune responses. Biomarkers reported by at least two trials were pooled using a random-effects model. Three RCTs (n = 196) met the eligibility criteria. hMSC therapy was associated with a significant reduction in tumor necrosis factor-α (TNF-α) at 6 months (mean difference [MD] −0.61, 95% CI −1.08 to −0.14), while interleukin (IL)-6, IL-8, interferon (IFN)-γ, and the CD4/CD8 ratio showed no consistent effects. Limited alloimmune data indicated no clinically meaningful immune sensitization. Overall, hMSC therapy may preferentially attenuate innate TNF-α–driven inflammation, while other immune effects remain exploratory.
Co-Authors Hendrata, Adi Pramono Kuncorojakti, Suryo Martha Ervina Novita, Bernadette Dian Sianty Dewi Sumi WIjaya, Sumi Suwandito, Laurensius Tjahjono, Yudy Wedharga, I Gede Putu Adhi Wijaya, Hendy