The Indonesian Biomedical Journal

106-23

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: Many chronic disorders, including vascular diseases, metabolic syndrome, and neurological diseases, are known to share a common factor of excessive inflammation. It takes a lot of energy to heal damaged tissue, which is a complicated process. The outcome is often suboptimal, with some degree of fibrosis, depending on the tissue's ability to regenerate and the strength of the inflammatory response. We may get new insights into disease causation and therapeutic strategies by better understanding endogenous regulatory points within the inflammatory response. CONTENT: Despite of the benefit in raising an inflammatory response, it also have unfavourable effects. Unresolved inflammation can over accumulate collagenous connective tissue and induce fibrosis, promote tissue dysfunction, and finally organ failure. Currently, the resolution of inflammation was described in terms of contemporary molecules as a different mechanisms from anti-inflammatory, since in resolution, the pathogen and apoptotic cells crumbs will be cleared and the macrophages will set back the tissue homeostasis. An active transition in the mediators that predominate in exudates occurs in conjunction with the remission of inflammation. These groups of inborn pro-resolution named resolvins, maresins, and protectins work to reduce inflammation by triggering certain pathways that support homeostasis rather than by suppressing the immune system. SUMMARY: The resolution of inflammation, once believed to be a passive process, is now understood to entail active biochemical programs that allow inflamed tissues to regain equilibrium. In this review, we spotlight the resolution to inflammation as a strategy to prevent tissue fibrosis and hinder the organ damage.

132-40

Authors: Aden Arrafif Bahtiarsyah, Lisna Hidayati, Nastiti Wijayanti, Tri Rini Nuringtyas

BACKGROUND: Type 2 diabetes mellitus affects glucose metabolism resulting in hyperglycemia. The bark of Cinnamomum burmannii (CB) and the leaves of Aquilaria malaccensis (AM) are believed to be effective for diabetes treatment. This study evaluated the synergistic effect of CB and AM, the extracts' phytochemical profiles, and the interaction between CB and AM metabolites with a-glucosidase through molecular docking. METHODS: The dried material was macerated with ethanol and then tested for a-amylase and a-glucosidase inhibitory activities and glucose diffusion inhibition in varied combination proportions. The extract fingerprinting was performed using a UV-Vis spectrophotometer followed by thin layer chromatography to determine the class of secondary metabolite in the extracts. Human maltase-glucoamylase (MGAM) receptor, ligands from acarbose and selected metabolites of CB and AM were studied in silico using UCSF Chimera, AutoDockTools, Autodock Vina Wizard (PyRx), and Biovia Discovery Studio software. RESULTS: The best ratio of CB:AM for a-amylase and a-glucosidase inhibition was 0.75:0.25 mg/mL, with inhibitory activities of 86.36 and 96.38 %, respectively. The best glucose diffusion inhibition was achieved at a ratio of 0.5:0.5 mg/mL CB and AM. The b-caryophyllene of CB and palustrol of AM had a significantly higher binding affinity of -10.7 kcal/mol and -10.2 kcal/mol, respectively than acarbose, which had a binding affinity of -8.1 kcal/mol. CONCLUSION: A ratio of CB to AM suppresses the activity of diabetes-related enzymes more efficiently. The in silico study suggested that the presence of b-caryophyllene in CB and palustrol in AM supported the synergistic activity.

150-6

Authors: Ahmad Syauqy Tafrihani, Nisa Ul Hasanah, Dhiya Ulhaq Salsabila, Ratih Kurnia Wardani, Ummi Maryam Zulfin, Muthi' Ikawati, Edy Meiyanto, Riris Istighfari Jenie

BACKGROUND: Cardamom (Amomum cardamom) essential oil (CEO) contains monoterpenes with antioxidant activity and is reported to exhibit anticancer activity against some cancer cell lines. Triple-negative breast cancer (TNBC) has the lowest prognosis among breast cancer types due to its aggressive characteristics. This study was conducted to explore the potency of CEO in inhibiting 4T1 cell proliferation and migration and compared its activity to sappan (Caesalpinia sappan) wood extract (CSE). METHODS: We used the 4T1 cell line as the TNBC cell model and tested the cytotoxicity of CEO by using a trypan blue exclusion assay. We studied the senescence induction ability of CEO using SA-β-Gal assay, the migratory inhibition activity using scratch wound healing assay, and inhibition of matrix metalloproteinase 9 (MMP-9) expression using gelatin zymography. RESULTS: CEO showed cytotoxicity toward 4T1 cells with the IC50 values of 60 µg/mL. CEO at ½ IC50 and IC50 concentration significantly increased cell senescence, but CSE did not. CEO at IC50 also reduced cell ability to migrate and also considerably reduced MMP-9 activity. Moreover, these activities related to the inhibition of the cell metastasis process were weaker compared than that of CSE. CONCLUSION: CEO showed potency as a chemopreventive agent on the TNBC 4T1 cell line model with moderate cytotoxicity. CEO induced 4T1 cell senescence, inhibited cell migration and suppressed MMP-9 expression.

165-70

Authors: Ratu Purwanti, Agnes Rengga Indrati, Nida Suraya, Maya Marinda Montain, Ines Atmosukarto, Neni Nurainy, Ida Parwati

BACKGROUND: BCG vaccine has been proven to be effective protection against tuberculosis (TB) meningitis and miliary TB. However, for protection against pulmonary TB, the results remains vary widely. Recombinant vaccine consisting of two immunodominant Mtb antigens ESAT-6-Ag85C-polyHistag (EAH) is currently being developed as a new TB vaccine candidate for booster. An immugonecity test for vaccine candidates is required in the initial phase to evaluate cellular immune response. This study was conducted to evaluate the cellular immune response by measuring interferon-gamma (IFN-γ) cytokines produced by T cells after ex vivo stimulation by TB EAH antigen fusion. METHODS: Peripheral blood mononuclear cells supernatant samples were collected from 16 new pulmonary TB subjects, 17 pulmonary TB in treatment subjects, and 10 healthy subjects. Samples were tested for IFN-γ level with enzyme-linked immunosorbent assay (ELISA). Kruskal-Wallis test was used to test the differences between IFN-γ levels among three groups, and followed by post-hoc analysis using Mann Whitney. RESULTS: The median of IFN-γ levels for new pulmonary TB, pulmonary TB in treatment, and healthy subjects were 17.09 (2.65-140.14) pg/mL, 4.36 (2.43-21.41) pg/mL, and 2.91 (2.39-3.85) pg/mL, respectively. There were significant differences of IFN-γ levels between new pulmonary TB group and pulmonary TB in treatment group (p=0.012), between new pulmonary TB group and healthy group (p=0.001), and also between pulmonary TB in treatment group and healthy group (p=0.035). CONCLUSION: Results show that TB EAH could stimulate cell-mediated immune responses in the three groups, with the highest IFN-γ levels are found in new pulmonary TB group, suggesting a potential immunodominant antigen fusion for vaccine candidate development.

179-86

Authors: Anastasia Armimi, Afina Firdaus Syuaib, Katherine Vanya, Marselina Irasonia Tan, Dessy Natalia, David Virya Chen, Chikako Ono, Yoshiharu Matsuura, Anita Artarini, Ernawati Arifin Giri-Rachman

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects humans' lower respiratory tracts and causes coronavirus disease-2019 (COVID-19). Neutralizing antibodies is one of the adaptive immune system responses that can reduce SARS-CoV-2 infection. This study aimed to develop a SARS-CoV-2 neutralization assay system using pseudo-lentivirus. METHODS: The plasmid used for pseudo-lentivirus production was characterized using restriction analysis. The gene encoding for SARS-CoV-2 spike protein was confirmed using sequencing. The transfection pseudo-lentivirus optimal condition was determined by choosing the transfection reagents and adding centrifugation step. Optimal pseudo-lentivirus infection was analysed using fluorescent assay and luciferase assay. The optimal condition of pseudo-lentivirus infection was determined by the target cell type and the number of pseudo-lentiviruses used for neutralization test. SARS-CoV-2 pseudo-lentivirus was used to detect neutralizing antibodies from serum samples. RESULTS: The plasmid used for pseudo-lentivirus production was characterized and confirmed to have no mutations. Lipofectamine 2000 reagent generated pseudo-lentivirus with a higher ability to infect target cells, as indicated by a percentage green fluorescent protein (GFP) of 12.68%. Pseudo-lentivirus centrifuged obtained more stable results in luciferase expression. Optimal pseudo-lentivirus infection conditions were obtained using puromycin-selected HEK 293T-ACE2 cells as target cells. The number of pseudo-lentiviruses used in the neutralization assay system was multiplicity of infection (MOI) 0.075. Serum A samples with a 1:10 dilution had the highest neutralizing antibody activity. CONCLUSION: This study shows that SARS-CoV-2 neutralization assay system using pseudo-lentivirus successfully detected neutralizing antibodies in human serum, which were indicated by a decrease in the percentage of pseudo-lentivirus infections.