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Corrigendum: Evaluating SARS-CoV-2 Spike Protein Transfection in HEK-293T Cells for VLP Applications Rohmah, Ilmiana Nurur; Marlita, Marlita; Kusuma, Kavana Hafil; Christina, Yuyun Ika; Dwijayanti, Dinia Rizqi; Mustikaningtyas, Dewi; Widodo, Nashi; Djati, M. Sasmito
The Journal of Experimental Life Science Vol. 15 No. 2 (2025)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2025.015.02.01

Abstract

The spike protein of SARS-CoV-2 is crucial for initiating infections by binding to host cells and mediating membrane fusion. In this study, HEK-293T cells were transfected with plasmids encoding three structural proteins of SARS-CoV-2, i.e., Spike (S), Membrane (M), and Envelope (E). This transfection enabled the formation of SARS-CoV-2 Virus-Like Particles (VLPs), which allows for safer studies of the virus and its proteins. To serve as a marker for expression, an enhanced green fluorescent protein (EGFP) was fused to the spike protein, resulting in a Spike-EGFP (S-EGFP) fusion protein. The characteristics of SARS-CoV-2 spike protein expression in the transfected HEK-293T cells were then investigated using polymerase chain reaction (PCR) and flow cytometry. The PCR analysis revealed non-specific DNA band smearing, which did not provide conclusive confirmation of spike protein expression. However, flow cytometry analysis demonstrated that approximately 30% of the transfected cells exhibited green fluorescence, indicating the expression of the Spike-EGFP fusion protein. These findings, obtained through flow cytometry, confirmed the successful spike protein expression in transfected HEK-293T cells.  
Determination of Polyphenol Compounds in Clinacanthus nutans and Their Potential as Antioxidants Widodo, Nashi; Salma Wahid Marseti; Yuslinda Annisa; Fairuz Sarah Kamila; Rosyadah, Nuraini; Rifa’i, Muhaimin
Biotropika: Journal of Tropical Biology Vol. 13 No. 1 (2025)
Publisher : Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.biotropika.2025.013.01.06

Abstract

Free radicals can cause oxidative damage if they are produced in excess amounts in the body. To prevent this, the work of endogenous antioxidants needs to be supported by safe and effective exogenous antioxidants. Clinacanthus nutans is one of the sources of natural polyphenol compounds that have the potential to be used as free radical-scavenging agents. The study began with the extraction of C. nutans simplicia with microwave assisted extraction (MAE) using a 96% ethanol solvent. The amounts of phenols and flavonoids in the extract was then measured using the Folin-Ciocalteu and aluminium chloride techniques, and their antioxidant activity was assessed using the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (NO) scavenging assays. Based on the results of the study, it is known that C. nutans ethanolic extract contains higher flavonoids (238.29 ± 10.87 mgQE.g-1) than the total phenols (148.76 ± 1.26 mgGAE.g-1). This study also showed the potential of C. nutans extract as a free-radical scavenger with an IC50 from the NO-scavenging and DPPH assays of 28.37 ± 3.38 ppm and 117.84 ± 3.93 ppm, respectively. Despite the presence of high polyphenol content in the extract, its antioxidant activity was found to be relatively weak. Consequently, further study is necessary to ascertain the specific class of compounds responsible for its antioxidant activity and to assess its effectiveness.
Daruju (Acanthus ilicifolius L.) May Exhibit Anti-Breast Cancer Activity Through Inhibition of Proliferation Regulators: A Computational Study: Daruju (Acanthus ilicifolius L.) as Anti-Breast Cancer Agent Rosyadah, Nuraini; Kamila, Fairuz Sarah; Hermanto, Feri Eko; Widyananda, Muhammad Hermawan; Grahadi, Rahmat; Dwijayanti, Dinia Rizqi; Widodo, Nashi; Ulfa, Siti Mariyah
Journal of Tropical Life Science Vol. 15 No. 1 (2025)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/

Abstract

Breast cancer's increasing prevalence globally underscores the urgent need for effective and gentle therapies, positioning the exploration of herbal remedies as a critical pursuit. Daruju (Acanthus ilicifolius L.) emerges as a compelling candidate due to its inherent bioactive components. This research pioneers the application of advanced computational techniques to unveil the latent anti-breast cancer potential within A. ilicifolius. Our in-silico investigation commenced by cataloging A. ilicifolius compounds using the KNApSAcK database and existing literature. These compounds underwent rigorous screening for drug-like characteristics via SWISS-ADME and potential biological activity using PASS ONLINE. Protein targets relevant to breast cancer were predicted through SWISS Target and the STRING database, integrated with Cytoscape for network visualization. Molecular docking, performed with PyRx 0.8, assessed the binding strength between the identified compounds and target proteins, with the most promising interactions selected for further scrutiny. The stability of these crucial interactions was then evaluated through molecular dynamics simulations using YASARA. This comprehensive computational strategy aims to pinpoint potential anti-breast cancer agents derived from A. ilicifolius. Initial analysis of 17 compounds from A. ilicifolius, based on chromatography, databases, and prior studies, narrowed down to five that adhered to Lipinski’s Rule of Five for drug-likeness: 4-O-beta-D-glucosyl-4-coumaric acid, (-)-lyoniresinol, α-amyrin, adenosine, and p-coumaric acid. These compounds were predicted to directly interact with key breast cancer-related proteins across pathways like estrogen signaling, JAK/STAT, and PI3K/AKT. Notably, molecular docking revealed strong binding affinities for α-amyrin with CDK4, ER, and EGFR (-7.5 kcal/mol, -9.5 kcal/mol, and -8.7 kcal/mol, respectively), comparable to known inhibitors. Molecular dynamics simulations further corroborated the stability of these complexes, analyzing RMSD and binding affinity parameters. Consequently, α-amyrin stands out as a promising anti-breast cancer agent within A. ilicifolius, exhibiting potential to inhibit proteins crucial for breast cancer cell proliferation and survival, including CDK4, ER, and EGFR.
Design, Construction and Expression of Spike Highly Conserved Region (HCR) SARS-CoV-2 and Cholera Toxin Subunit B Fusion Protein in Lactococcus lactis NZ3900: Construction of recombinant plasmid in Lactococcus lactis Kesuma, Suryanata; Adianingsih, Oktavia Rahayu; Winarsih, Sri; Widodo, Nashi; Yurina, Valentina
Journal of Tropical Life Science Vol. 15 No. 2 (2025): In Press
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/2xf1qp96

Abstract

Prevention of SARS-CoV-2 transmission has primarily been achieved through vaccination, which is generally administered via injection and may cause discomfort. No commercially available SARS-CoV-2 vaccines can be administered via the mucosal route. However, recent advancements have demonstrated that vaccination with Lactococcus lactis enables vaccine delivery through the mucosa. A promising target for SARS-CoV-2 vaccine materials is the highly conserved region (HCR) of the SARS-CoV-2 spike (SARS-CoV-2 HCR Spike). Vaccine efficacy is enhanced by adding Cholera Toxin Subunit B (CTB) as an adjuvant. HCR and CTB proteins were recombinantly fused using a synthetic gene with optimized codons. This study aimed to construct a fusion protein of the SARS-CoV-2 spike protein and CTB in L. lactis strain NZ3900. The construction and expression of fusion proteins were analyzed using sequencing and protein electrophoresis. Codon optimization resulted in a Codon Adaptation Index value of 0.93 and a GC content of 27.06%. The cloning results revealed the formation of L. lactis colonies expressing the Fusion protein of the SARS-CoV-2 HCR Spike and CTB, which formed yellow colonies on the selection Elicker medium. PCR and sequencing confirmed the presence of the hcr-ctb gene, with a length of 981 bp and 100% sequence similarity. The Fusion protein of the SARS-CoV-2 HCR Spike and CTB was successfully expressed with a molecular weight of >35 kDa. In conclusion, we successfully constructed a Fusion protein of the SARS-CoV-2 HCR Spike and CTB in L. lactis NZ3900 as a potential vaccine candidate for oral administration to prevent SARS-CoV-2 infection.
Hepatoprotection and Immunomodulation of Natural Killer and CD8 T Cells by Meniran-Turmeric Extract Combination in Mice Injected with 7,12-Dimethylbenz[a] Anthracene Puspitarini, Sapti; Widodo, Nashi; Widyarti, Sri; Azzamuddin, Haidar; Ulinnuha, Maria; Puspitoputri, Alya H.; Khansa, Shafanony; Rifa’i, Muhaimin
HAYATI Journal of Biosciences Vol. 33 No. 2 (2026): March 2026
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.33.2.499-507

Abstract

Combination herbal or polyherbal offers advantages by synergizing multiple plants to address health issues more effectively. Meniran and turmeric are traditionally used medicinal plants with notable pharmacological activities due to their bioactive compounds. This study investigated the curative effect of a meniran-turmeric extract combination on immunocompetent cells, particularly NK and CD8 cells, and on liver histopathology in mice administered 7,12-dimethylbenz[a]anthracene (DMBA). Female BALB/c mice (Mus musculus) were injected subcutaneously with DMBA at 45 mg/kg body weight (BW) in the mammary gland area and maintained for eight weeks. They were then treated with a 1:1 meniran-turmeric extract combination for two weeks at doses of 100, 300, 900, and 1,800 mg/kg BW, followed by liver and immune cell analyses. The results showed a lower CD8+ T-cell population in extract-treated groups (17.53±1.13%-19.79±1.51%) compared with the DMBA group (20.92±4.74%). Conversely, NK+ cell populations increased after extract treatment (0.33±0.01%-0.56±0.13%) compared with DMBA alone (0.27±0.02%). Liver histopathology indicated that extract combination administration did not cause more severe damage than that observed in the DMBA group. An extract combination of meniran and turmeric could modulate the immune system, particularly NK and CD8 cells, in mice injected with DMBA. In addition, the study indicated that the combination of meniran and turmeric extract might play a role as an antihepatotoxic agent. This research could serve as a consideration for future studies developing a combination formulation of meniran and turmeric.