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Hypertrophy Determination of H9c2 Cardiomyoblast Cell Line Using Wright-Giemsa Staining: An Experience in Developing an Acceptable and Easy-to-handle In-vitro Protocol Fadhillah, Muhamad Rizqy; Arozal, Wawaimuli; Wibowo, Heri; Bustami, Arleni; Sukmawati, Dewi; Kusmardi, Kusmardi; Triana, Novi; Khatimah, Nurul Gusti
Majalah Patologi Indonesia Vol. 33 No. 2 (2024): MPI
Publisher : Perhimpunan Dokter Spesialis Patologi Anatomik Indonesia (PDSPA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.55816/mpi.v33i2.682

Abstract

Introduction Cell-size area (CSA) becomes the standard parameter routinely tested in vitro for cardiac hypertrophy studies. Thus, staining is an essential tool for this purpose. As reported in a previous study, immunofluorescence staining is an established method for CSA. However, because it is expensive and requires a specialized microscope, e.g., immunofluorescence or confocal microscope, it is not applicable in a laboratory with limited equipment. Wright-Giemsa staining is a standard procedure in hematology laboratories and is inexpensive and convenient. Here, we shared our experience developing a CSA determination protocol using Wright-Giemsa in H9c2 cardiomyoblast. Methods The viability tests were performed on H9c2 to determine the effective dosage of angiotensin II and Irbesartan (standard drug). The H9c2 were divided into three groups: the control group (without either angiotensin II or irbesartan), the negative control (with angiotensin II), and the positive control (with angiotensin II and Irbesartan), triplicate for each group. The cells then are acclimatized overnight, serum-starved for one day, and incubated with angiotensin and irbesartan for 48 hours. Lastly, Wright-Giemsa was observed using a light microscope in three fields. The CSA was determined by three independent observers blindly, statistically different if the p<0.05 using ANOVA ways or Kruskal-Wallis. Results After the H9c2 induced by angiotensin-II 1 μM and Irbesartan 1μM, we found the CSA significantly differed among each group (p<0,0001). The negative control has a higher median and interquartile range (IQR) CSA (10.78 (6.79) um2) compared to the control group (median (IQR) 7.27 (4.91) um2) and positive control (median (IQR) 7.849(5.31) um2). Conclusion It can be concluded that the Wright-Giemsa might help determine the CSA for in-vitro hypertrophic studies.
Aktivitas Antiinflamasi Crude Extract Fukoidan dari Sargassum crassifolium pada Sel RAW 264.7 yang Diinduksi LPS Hikariastri, Pangartika; Winarno, Hendig; Kusmardi, Kusmardi; Laksmitawati, Dian Ratih; Abdillah, Syamsudin
Jurnal Kefarmasian Indonesia VOLUME 9, NOMOR 2, AGUSTUS 2019
Publisher : Pusat Penelitian dan Pengembangan Biomedis dan Teknologi Dasar Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22435/jki.v9i2.1547

Abstract

Inflammation is known as the basic mechanism underlying various chronic diseases. Macrophage activation by inflammatory stimulus induces the release of inflammatory mediators, thus the mediators becoming a promising target of anti-inflammatory drug development. Previous studies indicated that fucoidan has anti-inflammatory activity by inhibiting the release of proinflammatory mediators. The aim of this study is determining anti-inflammatory activity of fucoidan crude extract form Sargassum crassifolium Garut waters by observing its effect on proinflammatory cytokines TNF-α, IL-1β, dan IL-6. Fucoidan is a polysaccharide substance which has various characteristics, depending on the source and the extraction method which is influencing its bioactivity. Sargassum crassifolium collected from Garut-West Java is extracted using diluted HCl 0,1 M and precipitated with ethanol to obtain fucoidan crude extract. The crude extract is tested on LPS-induced RAW 264.7 cells to evaluate its effect on TNF-α, IL-1β, and IL-6 level using ELISA method. The result showed that fucoidan crude extract decreased the level of TNF-α by the dose of 25 and 50 μg/ml, and decreased the level of IL-1β and IL-6 by the dose of 25 μg/ml. The dose of 50 μg/ml failed to inhibit IL-1β and IL-6 production. This study showed that fucoidan crude extract derived from S. crassifolium has anti-inflammatory activity to RAW 264.7 cells by the dose of 25 μg/ml.
Potensi Ekstrak Etanol 96% Daun Salam (Syzigium polyanthum Wight. (Walp.)) terhadap Ekspresi p53 pada Sel Kanker HeLa Cell Lines Hidayati, Wahyu; Sjahid, Landyyun Rahmawan; Ismalasari, Wahyu; Kusmardi, Kusmardi
Jurnal Kefarmasian Indonesia VOLUME 10, NOMOR 2, AGUSTUS 2020
Publisher : Pusat Penelitian dan Pengembangan Biomedis dan Teknologi Dasar Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22435/jki.v10i2.1996

Abstract

Bay leaves (Syzigium polyanthum Wight. (Walp.) have been utilized as one of traditional medicines in Indonesian. However, the potency of the leaves for cancer treatment has not been well-explored. The aim of this study is to determine the expression of p53 tumor suppressor gene after administration of 96% ethanol extract of Bay leaves on HeLa cell lines. This study is begin by making extract using ethanol 96% as a solvent followed by phytochemical screening and extract evaluation of HeLa cells by immunohistochemistry. The observation of p53 protein expression is conducted using the Image J Profiler Plugin software 1.52a version. The H-score data is processed using the One-Way ANNOVA IBM SPSS 20 version program and continued with Post Hoc analysis using the Duncan test. The identification of 96% ethanol extract result showed that bay leaves contain flavonoids, alkaloids, saponins, phenols, terpenoids, and steroids. Based on the Post Hoc test results, the administration of 96% ethanol extract bay leaves with three dose variations (156 µg/ml, 234 µg/ml, 312 µg/ml) and negative control had significant differences, while the high dose (312 µg/ml) had not significant differences to positive control. The result showed that 96% ethanol extract of bay leaves by the dose of 312 ug/ml can inhibit the expression of mutant p53 protein in cervical cancer cells HeLa cell lines.
Phytochemical Constituents and Evaluation of Lipoxygenase Activity of Tetracera macrophylla Twigs Wall.ex Hook.f. & Thoms Ladeska, Vera; Elya, Berna; Hanafi, Muhammad; Kusmardi, Kusmardi
JSFK (Jurnal Sains Farmasi & Klinis) Vol 11 No 1 (2024): J Sains Farm Klin 11(1), April 2024
Publisher : Fakultas Farmasi Universitas Andalas

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.25077/jsfk.11.1.1-6.2024

Abstract

Lipoxygenase is an enzyme that catalyzes the formation of leukotriene, which is involved in various diseases  such as asthma, osteoporosis and atherosclerosis. Inhibition of this enzyme can overcome inflammatory diseases. Tetracera  macrophylla Wall.ex Hook. f.& Thoms plant has shown various biological activities, especially as an anti-inflammatory. This study  aims to analyze phytochemical constituents and evaluate the lipoxygenase activity of Tetracera macrophylla twig extract. The  sample was gradually macerated using n-hexane, ethyl acetate, and methanol. Phytochemical analysis of the extract was carried  out using the LC-MS/MS method. Lipoxygenase assay was performed in vitro using RAW 264.7 macrophage cells induced with  lipopolysaccharide using spectrofluorometry. As a comparison, zileuton, which has been established as a 5-LOX inhibitor and  quercetin, which has the potential as a 5-LOX inhibitor. The results showed that methanol extract of Tetracera macrophylla twigs (RM3) had lower specific LOX activity (0.350 mU/mL) than standard zileuton (7.927 mU/mL) and quercetin (12.837 mU/mL).  Decreased LOX activity can lessen the generation of leukotrienes, which cause inflammation. The study showed that methanol extract of Tetracera macrophylla twigs contains active compounds that can be developed as lipoxygenase inhibitors
Potensi Ekstrak Daun Ficus deltoidea Jack sebagai Anti Kanker Payudara Putri, Rizky Clarinta; Kusmardi, Kusmardi; Hakim, Rani Wardani
Majalah Kedokteran Indonesia Vol 74 No 1 (2024): Journal of The Indonesian Medical Association - Majalah Kedokteran Indonesia, Vo
Publisher : PENGURUS BESAR IKATAN DOKTER INDONESIA (PB IDI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.47830/jinma-vol.74.1-2024-1232

Abstract

Female breast cancer is the most common diagnosed cancer worldwide in 2020, with an estimated 2.3 million new cases or 11.7% from total 18,1 million cases, followed by lung, colorectal, prostate, and stomach cancers. In 2040, the breast cancer cases are predicted will be increased into 3 million cases per year or grow 40% from 2020 data. It is also estimated the number of deaths due to breast cancer will be one million death per year in 2040. In Indonesia, the number of new breast cancers reached 68,858 cases (16.6%) of a total of 396,914 new cases of cancer in 2020. ​ The incidence and mortality rates of breast cancer are 44 and 15.3 per 100,000 population, respectively in Indonesia. The most common breast cancer treatments used currently are surgery, chemotherapy, and radiation. These three treatments involve a rather invasive approach and often causes side effects. Serious side effects can reduce a patient's quality of life, lead to non-compliance to chemotherapy, and lead to drug resistance.​ Some cancer patients use alternative therapy such as complementary medicine including herbal medicine to treat the cancer. Patients try to discover and utilize natural-derived anti-cancer drugs because it believes has fewer side effects and more affordable. One of Indonesia plant which is therapeutically beneficial is Ficus deltoidea. Ficus deltoidea leaves had been used in traditional medicine to treat diabetes, inflammation, diarrhoea, cancer and infections. F. deltoidea leaves contained flavonoids, saponins, tannins, steroids and terpenes which known for its anticancer effect.
Comparison of c-MYC Expression between Patients with Germinal Center and Non-Germinal Center B-cell-like Diffuse Large B Cell Lymphoma Hanum, Sitti Fatimah; Hardjolukito, Endang Sri Roostini; Kusmardi, Kusmardi; Murtiani, Farida; Widiantari, Aninda Dinar; Setiawaty, Vivi
Indonesian Journal of Cancer Vol 19, No 2 (2025): June
Publisher : http://dharmais.co.id/

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33371/ijoc.v19i2.1277

Abstract

Background: c-MYC expression has been used as a prognostic marker to predict prognosis and determine therapeutic strategies in both Diffuse Large B Cell Lymphoma (DLBCL) subtypes. No study on c-MYC expression associated with DLBCL has ever been conducted in Indonesia. Our study aimed to evaluate differences in c-MYC expression in both DLBCL subtypes and assess the immunophenotype profile.Method: We selected 40 DLBCL cases and divided them into Germinal Center B-cell (GCB) and non-GCB subtypes using Hans Criteria. We evaluated c-MYC expression, and a cut-off value of 60.4% was determined using Receiver Operating Characteristic (ROC) curve analysis.Results: We found that c-MYC expression was significantly higher in GCB subtypes compared to non-GCB subtypes (n = 17 (42.5%) vs n = 20 (7.5%), p 0.000 and mostly had an immunophenotype of CD10+/BCL6+/MUM1+.Conclusion: Higher c-MYC expression is found more frequently in GCB subtypes. These findings suggest that c-MYC may play a subtype-specific role in DLBCL pathogenesis, potentially influencing therapeutic decisions for Indonesian patients. Future studies should validate these results in larger, multi-center cohorts and explore the mechanistic link between c-MYC and the GCB subtype and its clinical implications for targeted therapies.
Beetroot (Beta vulgaris) potential in preventing colorectal cancer using in-silico analysis Dwijayanti, Adisti; Azizah, Norma N.; Erlina, Linda; Kusmardi, Kusmardi; Ningsih, Sri S.; Fadilah, Fadilah; Hashim, Najihah M.
Narra J Vol. 5 No. 2 (2025): August 2025
Publisher : Narra Sains Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.52225/narra.v5i2.1578

Abstract

Colorectal cancer (CRC) is one of the leading causes of cancer-related mortality worldwide, necessitating the need for an effective therapeutic strategy. Beta vulgaris (beetroot) possesses active compounds that exert anti-cancer properties. The aim of this study was to evaluate the potential of beetroot as a preventative agent against the progression of CRC using differentially expressed gene (DEG) analysis and network pharmacology approaches. The protein-protein interaction network and molecular docking analyses were employed to assess the key interactions of beetroot active compounds with CRC-related target protein. Cytotoxicity of beetroot extract was experimentally evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay on the HT29 cell line. The result of this study showed that protein in the cell cycle was significantly enriched in CRC, with cyclin-dependent kinase 4 (CDK4) gene as one of the specific genes. Quercetin, galangin, hesperidin, farrerol, and betanin were the most typical compounds of beetroot based on the Comparative Toxicogenomics Database (CTD). Molecular docking studies revealed the strong binding affinity between quercetin (-7.04 kcal/mol) and bentanin (-8.11 kcal/mol) with CDK4. Beetroot demonstrated anticancer properties against the HT29 cell line with IC50 value of 39.03±1.4 µg/mL. In conclusion, the beetroot extract has inhibitory activity against HT29 cell line proliferation, highlighting its potential in preventing the development of CRC through the substantial suppression of gene expression within the cell cycle pathway.
Hipertiroid dan Badai Tiroid: Tinjauan Pustaka Nina Yolanda Putri; Ariadi Humardhani; Kusmardi, Kusmardi
JURNAL RISET RUMPUN ILMU KEDOKTERAN Vol. 4 No. 3 (2025): Desember : Jurnal Riset Rumpun Ilmu Kedokteran
Publisher : Pusat riset dan Inovasi Nasional

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.55606/jurrike.v4i3.6984

Abstract

Hyperthyroidism is an endocrine disorder characterized by excessive production of thyroid hormones, with Graves' disease being the most common cause. This condition can lead to serious complications, including thyroid crisis or storm, which is a medical emergency with a high mortality risk. This article aims to review the literature related to the definition, epidemiology, etiology, clinical manifestations, pathophysiology, diagnosis, management, complications, and prognosis of hyperthyroidism and thyroid crisis. The writing method used a narrative literature review based on national and international literature from 2014 to 2024 obtained through PubMed, ScienceDirect, Google Scholar, and Sinta-indexed national journals. The results of the review show that hyperthyroidism has a broad impact on organ systems, with cardiovascular and metabolic complications being the main burden. Thyroid crisis, although rare, remains a serious threat that requires prompt, aggressive, and multidisciplinary management. In conclusion, early detection, accurate diagnosis, and guideline-based therapy are key to improving patient prognosis and reducing mortality.
The Effect of an Annona muricata Leaves Extract on Circulating Soluble Adhesion Molecules in Colon Carcinogenesis Model Indrawati, Lili; Kusmardi, Kusmardi; Wiyanto, Marwito
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy Vol 24, No 1 (2023): VOLUME 24, NUMBER 1, April, 2023
Publisher : The Indonesian Society for Digestive Endoscopy

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24871/241202330-34

Abstract

Background: Annona muricata leaves are used as traditional tea drink that is currently being studied on the developing effort of treating some types of cancers because they are known as anti-cancer. Motivated by that usage, this study aims to analyse the potential of A. muricata leaves extract to be an anti-colon cancer by investigating the extract capability in reducing blood intercellular cell-adhesion molecule-1 (ICAM-1) and vascular cell-adhesion molecule-1 (VCAM-1).Method: The research was conducted in Faculty of Medicine Universitas Indonesia with an ex­perimental design. A. muricata leaves extract was tested in vivo. In vivo test was conducted to Swiss Webster mice induced with 10 mg/kg azoxymethane (AOM) and dextran sodium sulfate 1% (DSS). Statistical analysis used was SPSS.Results: EIFAM reduce the serum level of ICAM-1 and VCAM-1. EIFAM significantly reduce serum level of VCAM compare to ESFAM.Conclusion: Ethanol insoluble fraction of Annona muricata leaves water extract of is potential to be an anti-colon cancer proven by the extract capability to reduce ICAM-1 and VCAM-1.
Effects of Sambiloto (Andrographis paniculata) Extract and Spirulina (Spirulina platensis) Administration on Ki-67 Protein Expression in the Colon Epithelial Cells of Plasmodium berghei-infected Mice Kusmardi, Kusmardi; Ariffandi, Bagas; Lubis, Nadar Sukri; Lestari, Tri Wahyuni; Intan, Putri Reno; Pakpahan, Alfred
The Indonesian Biomedical Journal Vol 14, No 1 (2022)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v14i1.1737

Abstract

BACKGROUND: Malaria remains a global health concern and an endemic disease in Indonesia. Sambiloto (Andrographis paniculata) and spirulina (Spirulina platensis) are two potential antimalarial agents which act as antioxidants and antiinflammatories that can suppress morbidities during chronic inflammation in malaria, such as proliferation in the colon. This study aims to investigate the effects of A. paniculata extract and S. platensis administration on Ki-67 expression in medial colon epithelial cells of Plasmodium berghei-infected mice measured by H-score.METHODS: Thirty P. berghei-infected male Swiss-Webster mice were divided into five groups: negative controls (carboxymethyl cellulose/CMC); positive controls (dihydroartemisinin-piperaquine/DHP); A. paniculata extract alone (AP); A. paniculata extract in combination with S. platensis extract (AP+ES); and with S. platensis powder (AP+PS). All mice were infected with P. berghei on day 0. The treatment for each group were given 3 days before infection (D-3) until the day of infection (D0) for 28 days after infection. Colon tissues were processed with immunohistochemistry to detect Ki-67.RESULTS: A difference in Ki-67 expression was observed among the groups (p<0.01). The mean H-score for the CMC control group is 135.503±6.723. The lowest level of Ki-67 expression was observed in the AP+PS group (H-score= 110.941±7.079). AP group did not show a significant difference from the CMC group (p=0.514) and neither did the AP+ES group (p=0.234).CONCLUSION: In conclusion, administration of A. paniculata extract and S. platensis powder lowers Ki-67 expression in medial colon epithelial cells of P. berghei-infected mice.KEYWORDS: malaria, spirulina, Ki-67, Andrographis paniculata, Spirulina platensis
Co-Authors . Syamsudin Adisti Dwijayanti Alfred Pakpahan, Alfred Aniati Murni Anwar S. Ibrahim Anwar S. Ibrahim Aprilliyani, Ifana Ariadi Humardhani Ariffandi, Bagas Arleni Bustami Aryo Tedjo Athifa, Khasna Fayza Atmaja, Fredy Wirya Azizah, Norma N. Berna Elya Budiana Tanurahardja Dewi Sukmawati Diah Rini Handjari Dian Ratih Laksmitawati Dimas Ramadhian N Ditha Diana Ditha Diana Dwira, Surya Dwitia Wulandari Endang SR. Hardjolukito Endang Sri Roostini Hardjolukito Enif Esti Triana Ening Krisnuhoni Erlina, Linda Erwina Muhadi Fadhillah, Muhamad Rizqy Fadilah Fadilah, Fadilah Fajriati, Annisa Hamdani Zain Hanum, Sitti Fatimah Hashim, Najihah M. Hayun Hayun, Hayun Hendig Winarno Heri Wibowo Heru Suhartanto Hikariastri, Pangartika Imelda Setiana Silitonga Irandi Putra Pratomo Ismalasari, Wahyu Jasni Mohammad Zain Khatimah, Nurul Gusti Kristanty, Diyah Ladeska, Vera Landyyun Rahmawan Sjahid Landyyun Rahmawan Sjahid Lestari, Tri Wahyuni Lili Indrawati Lubis, Nadar Sukri Megawati, Ajeng Muhammad Hanafi Murtiani, Farida Nina Yolanda Putri Noor, Dimas Ramadhian Novi Fajar Utami Nurdin, Wildawati Peni Kistijani Samsuria Putri Reno Intan Putri, Rizky Clarinta Rani Wardani Hakim Rino Patiatta Rohmat, Siti Saudah Shirly Kumala Sibli, Sibli Sri S. Ningsih, Sri S. Sri Suryo Adiyanti, Sri Suryo Surya Dwira Syamsu Nur, Syamsu Toto Haryanto Triana, Novi Virhan Novianry Vivi Setiawaty Wahyu Hidayati Wahyu Hidayati Wahyu Ismalasari Wawaimuli Arozal Widiantari, Aninda Dinar Wiwiek Ernajanti Wiyanto, Marwito Yulhasri Yulhasri Yusoff, Marina Yusra Yusra