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The effect of Phyllanthus niruri and Catharanthus roseus on Macrophage Polarization in Breast Cancer Mice Model: The Effect of P. niruri and C. roseus in Breast Cancer Mice Model Sakti, Sefihara Paramitha; Sari, Fikriya Novita; Rachmawati, Farida; Widyarti, Sri; Rahayu, Sri; Soewondo, Aris; Jatmiko, Yoga Dwi; Rifa'i, Muhaimin
Journal of Tropical Life Science Vol. 14 No. 1 (2024)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.14.01.03

Cancer death cases have increased yearly, and there are estimated to be 21.6 million cancer cases in 2030. Studies of herbal compounds for cancer treatment alternatives are essential because cancer treatment is relatively expensive and has adverse effects. Phyllanthus niruri (Pn) and Catharanthus roseus (Cr) are plants that are known as herbal medicines. Combining the two plants is expected to prevent and enhance the immune system in breast cancer cases. This study aims to analyze the anti-cancer and immunomodulatory effects of P. niruri and C. roseus extract (PCE) in modulating macrophage polarization in breast cancer mice. Experimental animals are divided into six groups and there is healthy control (normal mice), cancer (DMBA-induced mice), cancer mice with cisplatin administration, cancer mice with PCE administration with three different doses, including dose 1 (500 mg/kg Pn + 15 mg/kg Cr), dose 2 (1000 mg/kg Pn + 75 mg/kg Cr), and dose 3 (2000 mg/kg Pn + 375 mg/kg Cr). The mice were injected with DMBA once a week for six weeks to induce cancer in mice. The breast cancer mice model was administered with PCE orally for 14 days. The expression of CD11b+IL-10+ and CD11b+IFN-γ+ demonstrated macrophage polarization. The results showed that breast cancer induction using DMBA increased the level of IL-10 and decreased the level of IFN-γ significantly compared to the normal group (p < 0.05). In specific doses, administration of PCE could reduce IL-10 levels and increase the level of IFN-γ significantly (p < 0.05). PCE can modulate the polarization of macrophages by suppressing the M2-like macrophage and increasing the M1-like macrophage. The ability of PCE to modulate macrophage polarization indicates that the combination of P. niruri and C. roseus has activity as an anti-cancer.

Effect of Combination of Different Antibiotics and Promoters for Expressing Recombinant Darbepoetin in Stable CHO K-1 Cell Line: Evaluating Antibiotic Combinations to Improve Yield and Quality of Darbepoetin Widekdo, Dwi purno; Widodo, Nashi; Rifa'i, Muhaimin; Jatmiko, Yoga Dwi
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/

Antibiotics are key for successful molecular cloning techniques. Different antibiotics have different mechanisms of action, which leads to cell heath and a viable number of passages. Moreover, the suitability of the promoter also plays an important role in achieving a higher level of protein titter in the stable cell platform. Therefore, with plenty of options of antibiotics and promoters available, we need to determine the best combination of antibiotics and promoters, particularly for specific proteins of interest. Darbepoetin is a recombinant therapeutics protein with extra glycosylation to increase the half-life in the blood; this drug is used for the administration of CKD and leukemia patients. Blasticidin-S and puromycin were used as antibiotics, and CMV and EF-1 promoters were used in this experiment to evaluate the expression of recombinant darbepoetin for the protein model. CHO K-1 cell line was transfected with a plasmid carrying a combination of promoter and antibiotics genes; after 14 days, the level of specific protein expression was evaluated using the western blot technique. A single clone cell was obtained using the serial dilution method to evaluate the clonality and expression of the protein of interest. This study successfully obtained a single clone from stable pool transfection. This result suggested that a combination of puromycin antibiotics and EF-1 promoter has promising expression compared to Blasticidin-S antibiotics with CMV promoter. For further conclusion, an analytical comparison of both combinations needed to be done.

The effect of Phyllanthus niruri and Catharanthus roseus on Macrophage Polarization in Breast Cancer Mice Model: The Effect of P. niruri and C. roseus in Breast Cancer Mice Model Sakti, Sefihara Paramitha; Sari, Fikriya Novita; Rachmawati, Farida; Widyarti, Sri; Rahayu, Sri; Soewondo, Aris; Jatmiko, Yoga Dwi; Rifa'i, Muhaimin
Journal of Tropical Life Science Vol. 14 No. 1 (2024)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.14.01.03

Cancer death cases have increased yearly, and there are estimated to be 21.6 million cancer cases in 2030. Studies of herbal compounds for cancer treatment alternatives are essential because cancer treatment is relatively expensive and has adverse effects. Phyllanthus niruri (Pn) and Catharanthus roseus (Cr) are plants that are known as herbal medicines. Combining the two plants is expected to prevent and enhance the immune system in breast cancer cases. This study aims to analyze the anti-cancer and immunomodulatory effects of P. niruri and C. roseus extract (PCE) in modulating macrophage polarization in breast cancer mice. Experimental animals are divided into six groups and there is healthy control (normal mice), cancer (DMBA-induced mice), cancer mice with cisplatin administration, cancer mice with PCE administration with three different doses, including dose 1 (500 mg/kg Pn + 15 mg/kg Cr), dose 2 (1000 mg/kg Pn + 75 mg/kg Cr), and dose 3 (2000 mg/kg Pn + 375 mg/kg Cr). The mice were injected with DMBA once a week for six weeks to induce cancer in mice. The breast cancer mice model was administered with PCE orally for 14 days. The expression of CD11b+IL-10+ and CD11b+IFN-γ+ demonstrated macrophage polarization. The results showed that breast cancer induction using DMBA increased the level of IL-10 and decreased the level of IFN-γ significantly compared to the normal group (p < 0.05). In specific doses, administration of PCE could reduce IL-10 levels and increase the level of IFN-γ significantly (p < 0.05). PCE can modulate the polarization of macrophages by suppressing the M2-like macrophage and increasing the M1-like macrophage. The ability of PCE to modulate macrophage polarization indicates that the combination of P. niruri and C. roseus has activity as an anti-cancer.

Development of a Stable CHO Cell Line for High-Level Expression and Purification of Recombinant Darbepoetin Alfa Widekdo, Dwi purno; Widodo, Nashi; Rifa'i, Muhaimin; Jatmiko, Yoga Dwi
Journal of Tropical Life Science Vol. 15 No. 3
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.15.03.12

Recombinant Darbepoetin alfa (DARB), an erythropoiesis-stimulating agent, is widely used in treating anemia linked to chronic kidney disease and chemotherapy. This study presents a systematic approach for developing a high-yield, stable Chinese hamster ovary (CHO) cell line capable of producing recombinant DARB. A codon-optimized gene construct under dual CMV/EF1α promoters and puromycin resistance selection was transfected into CHO cells. To isolate high-producing monoclonal populations, transfected cells were subjected to limiting dilution cloning in 96-well plates, allowing single-cell-derived colonies to expand and be individually assessed. Clone DARV IV Pool 2 Clone 2E6 demonstrated the highest and most consistent DARB expression, validated through Western blot using anti-human EPO antibodies. The culture supernatant underwent two-step clarification via centrifugation and tangential flow filtration (TFF), followed by purification using anion-exchange chromatography on a HiTrap Q HP column. Gradient elution enabled effective separation, with SDS-PAGE and Western blot confirming high purity and molecular integrity of the recombinant protein. Variability in clone expression highlighted the influence of genomic integration sites and potential epigenetic silencing, emphasizing the importance of screening and stability validation. This study demonstrates that rational vector design, antibiotic-based clone selection, and robust purification strategies can produce CHO-derived DARB suitable for large-scale production. The integrated workflow supports scalability, product consistency, and regulatory readiness for biosimilar therapeutic manufacturing

Phenotypic and Genotypic Characterization of Tetracycline-Resistant Bacteria Isolated from Brantas River, Malang Raya and Their Biofilm Formation Ability: Phenotypic and Genotypic Characterization of Tetracycline-Resistant Bacteria Mahira, Syarafin; Yoga Dwi Jatmiko; Maharani Pertiwi Koentjoro
Journal of Tropical Life Science Vol. 16 No. 01 (2026)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.16.01.14

Antibiotic resistance in aquatic environments has become an important public health concern due to the spread of antibiotic-resistant bacteria (ARB) through river water, which provides favorable conditions for biofilm formation and enables bacteria to survive environmental stresses, including antibiotic exposure. The Brantas River water is still widely used for household activities, increasing the potential dissemination of bacteria through water exposure. This study aims to determine the phenotypic characteristics, identify tetracycline-resistant bacterial species using 16S rRNA gene sequencing, and analyze the level of biofilm formation and cell density within the biofilm by tetracycline-resistant bacterial isolates. Tetracycline-resistant isolates were phenotypically characterized and identified using 16S rRNA gene sequencing. Detection of tetracycline resistance genes was performed by Polymerase Chain Reaction (PCR). Biofilm formation ability was evaluated using the crystal violet (CV) assay in Tryptic Soy Broth (TSB) and Brantas River water, exopolysaccharide (EPS) extraction, and then the cell density in the biofilm was calculated using Total Plate Count (TPC). Isolates TET-15.1 and TET-40.1 were found to be resistant to all four concentrations. Identification results showed similarities between TET-15.1 with Alcaligenes faecalis; and TET-40.1 with Serratia sp. The resistant genes found in both isolates were tet(S) and tet(B). Alcaligenes faecalis TET-15.1 had the highest biofilm formation ability and higher EPS production than Serratia sp. TET-40.1, with OD584 values ranging from 1.5 to 2.25 and EPS concentration of 4 – 5 mg/mL at tetracycline concentration of 8 μg/mL. The biofilm formation cycle reached maturity after 16 hours of incubation and then dispersed after 32 hours. These findings indicate that tetracycline-resistant bacteria from the Brantas River possess biofilm forming capabilities that may contribute to their environmental persistence and potential role in the dissemination of antibiotic resistance.

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