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Journal : Medical Journal of Indonesia

Expressions of stemness markers in keloid tissue Ningsih, Sri S.; Sari, Dewi H.; Antarianto, Radiana D.; Hardiany, Novi S.; Sadikin, Mohamad; Wanandi, Septelia I.; Jusman, Sri W.A.
Medical Journal of Indonesia Vol 27, No 3 (2018): September
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (280.506 KB) | DOI: 10.13181/mji.v27i3.1920

Abstract

Background: Keloid is an abnormal wound healing process that extends beyond the site of injury. Keloid and tumor’s shared similarity of recurrence suggesting a shared underlying mechanism that involves stemness. Octamer-binding transcription factor-4 (Oct-4) and aldehyde dehydrogenase-1 (ALDH1) are stem cell stemness markers. This study aimed to analyze Oct-4 and ALDH1 expressions in keloid tissues.Methods: Samples were obtained from keloid tissue excisions from three keloid patients and post-circumcision preputial skin from three healthy donors (normal control) in accordance with the local ethical committee regulation. Total RNA was isolated using TriPure Isolation kit (Ameritech), and expressions of Oct4 and ALDH1 mRNA in keloid and preputial skin were determined by quantitative reverse transcription–polymerase chain reaction (qRT-PCR) using Livak method.Results: The qRT-PCR analysis revealed the expressions of Oct4 and ALDH1 in keloid and preputial skin tissues. Keloid tissues exhibited lower expression levels of Oct-4 and ALDH1 than the preputial skin. The difference was statistically insignificant.Conclusion: Keloid tissues express Oct-4 and ALDH1 as stemness markers, and the stemness characteristics of keloid might be similar to a normal skin.
Expression and specific activities of carbamoyl phosphate synthetase 1 in chronic hypoxic rats Nikmah, Uly A.; Prijanti, Ani R.; Jusman, Sri W.A.; Sadikin, Mohamad
Medical Journal of Indonesia Vol 25, No 1 (2016): March
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (430.303 KB) | DOI: 10.13181/mji.v25i1.1213

Abstract

Background: Urea biosynthesis is a very important process in the liver which needs ATP, CO2 and functional mitochondria or aerobic condition. Liver can adapt to hypoxic condition, generally and locally. This study aimed to analyze the effect of chronic hypoxia on liver urea biosynthesis as indicated by the level and specific activity of mRNA of carbamoyl phosphate synthetase 1 (CPS1), a key enzyme in urea biosynthesis in hypoxic rats.Methods: 20 male Sprague-Dawley rats were placed in hypoxic chamber supplied by a mixture of 10% O2 and 90% N2. Five rats were sacrificed at 1, 3, 5, and 7 days after exposure. Liver homogenates were analyzed for HIF-1 (hypoxia inducible factor-1) by ELISA, CPS1 mRNA by real time RT-PCR and CPS1 enzymatic specific activities by Pierson method. Data were analyzed by ANOVA test and Pearson correlation.Results: The HIF-1 in liver increased significantly, as well as CPS1 mRNA and CPS1 enzymatic activities (p<0.05). There was a strong correlation (r=0.618; p<0.01) between the level of CPS1 mRNA and CPS1 enzymatic activities, moderate correlation between HIF-1 and CPS1 mRNA (r=0.419; p<0.05) but no correlation between HIF-1 and CPS1 enzymatic activities. The study indicated that urea biosynthesis in liver was affected by hypoxia and partially under HIF-1 regulation. The study also found increase of urea and NH3 biosynthesis related to proteolysis as indicated by the decrease of total body weight and liver weight.Conclusion: There was an increase in the expression and specific activities of CPS1 in urea biosynthesis as a result of increasing proteolysis  in chronic hypoxic condition.
Myocardial damage after continuous aerobic and anaerobic exercise in rats Flora, Rostika; Ferdinal, Frans; Hernowo, Bethy S.; Wanandi, Septelia I.; Sadikin, Mohamad; Freisleben, Hans-Joachim
Medical Journal of Indonesia Vol 22, No 4 (2013): November
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (813.04 KB) | DOI: 10.13181/mji.v22i4.601

Abstract

Background: Regular physical activity is highly recommended in preventive, curative, and rehabilitative programs in order to promote health, especially cardiovascular health. However, physical activity can also cause sudden death. In athletes, sudden death may occur during sport competitions, with myocardial infarction as the most common etiology. It is suspected that continuous training without any rest-day play a role in cardiac muscle damage and sudden death during competition. Our study was aimed to learn about cardiac muscle adaptation on continuous aerobic and anaerobic physical activity without any rest-day. Methods: The specimens in our study were cardiac muscle tissue obtained from rats that had performed aerobic and anaerobic physical activity on treadmill for 1, 3, 7, and 10 days without any rest-day. Blood gas analysis and hematological assessment were used as parameters of systemic adaptation to hypoxia during physical activity. Moreover, histopathology of cardiac muscle tissue was performed as parameter for cardiac muscle damage.Results: The results showed that aerobic and anaerobic physical activity caused a systemic hypoxic condition and triggered adaptation responses. Cardiac muscle damage occurred on the 10th day in both treatment groups, with more severe damage observed in the group with anaerobic physical activity. The tissue protein level in the anaerobic group increased progressively on the 10th day.Conclusion: Physical activity may result in hypoxia and systemic adaptation. Aerobic and anaerobic physical activities performed for 10 days without any rest-day may cause cardiac muscle damage. (Med J Indones. 2013;22:209-14. doi: 10.13181/mji.v22i4.601)Keywords: Cardiac muscle, cardiac muscle damage, histopathology, physical activity
The suppression of manganese superoxide dismutase decreased the survival of human glioblastoma multiforme T98G cells Hardiany, Novi S.; Sadikin, Mohamad; Siregar, Nurjati; Wanandi, Septelia I.
Medical Journal of Indonesia Vol 26, No 1 (2017): March
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1015.302 KB) | DOI: 10.13181/mji.v26i1.1511

Abstract

Background: Glioblastoma multiforme (GBM) is a primary malignant brain tumor which has poor prognosis. High incidence of oxidative stress-based therapy resistance could be related to the high antioxidant status of GBM cells. Our previous study has reported that manganese superoxide dismutase (MnSOD) antioxidant expression was significantly higher in high grade glioma than in low grade. The aim of this study was to analyze the impact of MnSOD suppression toward GBM cell survival.Methods: This study is an experimental study using human glioblastoma multiforme T98G cell line. Suppression of MnSOD expression was performed using in vitro transfection MnSOD-siRNA. The MnSOD expression was analyzed by measuring the mRNA using real time RT-PCR, protein using ELISA technique, and specific activity of enzyme using inhibition of xantine oxidase. Concentration of reactive oxygen species (ROS) intracellular was determined by measuring superoxide radical and hydrogen peroxide. Cell survival was analyzed by measuring viability, proliferation, and cell apoptosis.Results: In vitro transfection of MnSOD-siRNA suppressed the mRNA, protein, and specific activity of MnSOD. This treatment significantly increased the concentration of superoxide radical; however, it did not influence the concentration of hydrogen peroxide. Moreover, viability MnSOD-suppressing cell significantly decreased, accompanied by increase of cell apoptosis without affecting cell proliferation.Conclusion: The suppression of MnSOD expression leads to decrease glioblastoma multiforme cell survival, which was associated to the increase of cell apoptotic.
Avidin inhibits PHA-induced human peripheral blood mononuclear cell proliferation Firakania, Cicia; Mansur, Indra G.; Jusman, Sri W.A.; Sadikin, Mohamad
Medical Journal of Indonesia Vol 25, No 1 (2016): March
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1031.939 KB) | DOI: 10.13181/mji.v25i1.1264

Abstract

Background: Cell proliferation occurs not only in normal but also in cancer cells. Most of cell proliferation inhibition can be done by inhibiting the DNA synthesis, notably by intervening the formation of purine or pyrimidine. In purine de novo synthesis, it was assumed that biotin plays a role as a coenzyme in carboxylation reaction, one of the pivotal steps in the purine de novo pathways. The aim of this study was to see the avidin potency to bind biotin and inhibit mitosis.Methods: Peripheral blood mononuclear cell (PBMC) was cultured in RPMI-1640 medium and stimulated by phytohemagglutinin (PHA) in the presence or absence of interleukin-2 (IL-2), with or without avidin. The effect of avidin addition was observed at 24, 48, and 72 hours for cell proliferation, viability, and cell cycle. Statistical analysis was done by one-way ANOVA.Results: Avidin inhibited cell proliferation and viability in culture under stimulation by PHA with and without IL-2. Cell cycle analysis showed that avidin arrested the progression of PBMC after 72 hours of culture. Most cells were found in G0/G1 phase.Conclusion: Inhibition of biotin utilization by avidin binding can halt cell proliferation.
Relative hypoxia and oxidative stress in spleen lymphocytes of immunized Balb/c mice as indicated by HIF-1α, HIF-2α, Nrf2 expression, and glutathione peroxidase activity Praditi, Citra; Prijanti, Ani R.; Jusman, Sri W.A.; Sadikin, Mohamad
Medical Journal of Indonesia Vol 27, No 4 (2018): December
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (410.853 KB) | DOI: 10.13181/mji.v27i4.2152

Abstract

Background: Lymphocytes activated by immunization must increase their metabolism to meet the energy requirements for mitosis, differentiation, and protein synthesis, which may subject the cell to conditions of relative hypoxia and oxidative stress. This study was conducted to investigate the increase in the levels of transcription factors involved in both conditions.Methods: Male Balb/c mice were divided into the following four groups, each consisting of six animals: the control and three experimental groups. The experimental groups were immunized by injection of 0.2 ml of 2% sheep red blood cells (SRBC) suspended in phosphate-buffered saline (PBS). Lymphocytes were harvested from the spleens of each group at time intervals of 24-, 48-, and 72-h post-immunization. The buffy coat from splenocytes was separated using Ficoll Histopaque as the medium. The lymphocytes were separated from adherent cells by incubating the purified splenocytes in microtubes for 2-h. Cells were lysed by three freeze–thaw cycles (−80°C and 37°C) and used to analyze the levels of HIF-1α and HIF-2α (mRNA and protein), Nrf2 (protein), and glutathione peroxidase (GPx) activity.Results: The treatment caused an increase in GPx activity and HIF-1α protein concentration 24-h post-immunization, whereas the HIF-1α mRNA levels remained static. Elevated Nrf2 protein levels were detected within 48-h after treatment. Meanwhile, the HIF-2α mRNA and protein levels increased within72-h after immunization.Conclusion: Immunization with SRBC suspension induced relative hypoxia, elevated reactive oxygen species (ROS), and oxidative stress in the lymphocytes as indicated by the increase in both HIF-1α and HIF-2α protein and mRNA levels, GPx activity, and Nrf2 protein levels.
Efforts to overcome hypoxia condition in Balb/c mouse macrophages after intraperitoneal SRBC immunization Sarsanti, Pungguri Ayu Nega; Sadikin, Mohamad; Jusman, Sri Widia Azraki
Medical Journal of Indonesia Vol 28, No 1 (2019): March
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (836.675 KB) | DOI: 10.13181/mji.v28i1.1961

Abstract

BACKGROUND Activated macrophages require increased oxygen to destroy foreign bodies, leading to an increase in the levels of reactive oxygen species (ROS). Therefore, macrophages would experience hypoxic and oxidative stress conditions at the same time. Thus, this study was aimed to evaluate the mechanism of the activated macrophages to overcoming this dual condition.METHODS The activated macrophages were harvested from the intraperitoneal cavities of 18 BALB/c mice immunized with 2% sheep red blood cells (SRBCs). The macrophage suspension was divided into four groups: control, 24, 48, and 72 hours after-immunization groups. The expressions of hypoxia-inducible factor (HIF)-1α, HIF-2α, and cytoglobin (Cygb), as markers for hypoxic condition, were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA), whereas peroxisome proliferator-activated receptor gamma coactivator (PGC-1α) protein as a marker for mitochondrial biogenesis and aerobic metabolism was measured with ELISA. The analysis of oxidative stress was conducted with the water-soluble tetrazolium salt test.RESULTS The HIF-1α mRNA expression was the highest at 24 hours, whereas the HIF-2α mRNA showed no increased expression during the observation. The Cygb mRNA decreased after 24 hours. The highest expressions of HIF-1α and HIF-2α proteins were detected at 72 hours, whereas the Cygb protein expression increased since 24 hours. The PGC-1α protein expression increased at 72 hours. The WST test showed the highest ROS level at 24 hours.CONCLUSIONS The macrophages were activated by SRBCs underwent dual hypoxia and oxidative stress condition simultaneously to overcome the foreign bodies. The macrophages overcame these stress conditions by increasing their aerobic metabolism.
Co-Authors . Rusdi Ahmad A. Jusuf Ahmad Aulia Ahmad Aulia Jusuf Ahmad R. Utomo Amarila Malik AMARILA MALIK Andi N.K. Syarifin, Andi N.K. ANDREW ROBERT COSSINS Angelina Stevany Regina Masengi Ani R. Prijanti Arief Budi Witarto Aryenti . Asri Rasad Astutiati Nurhasanah ASTUTIATI NURHASANAH Bethy S. Hernowo Caroline T. Sardjono CAROLINE TAN SARDJONO Cicia Firakania, Cicia DARYL ROBERT WILLIAMS Dewi Sukmawati Diah Iskandriati Dian R. Laksmitawati Dian Ratih Laksmitawati Dwirini Retno Gunarti Ekawati, Maria Erni Hernawati Purwaningsih, Erni Hernawati Fanny Septiani Farhan Febriana C. Iswanti Febriana Catur Iswanti Franciscus D. Suyatna Frans Ferdinal HANIFAH RAHMI Hans -Joachim Freisleben Hans Joachim Freisleben HANS-JOACHIM FREISLEBEN Heri Wibowo Hermansyah Hermansyah Indra G. Mansur Jeanne A. Pawitan JEANNE ADIWINATA PAWITAN Juniarti . Jusman, Sri Widia Azraki Lailan Safina Nasution Lio, Tiara Mayang Pratiwi Mohamad Saekhu Muchlis Ramli Nani Dharmasetiawani Nina Fitriana, Nina Ninik Mudjihartini Noorwati Sutandyo Novi S. Hardiany Novi Silvia Hardiany Nurhadi Ibrahim Nurjati C. Siregar Oentoeng Soeradi Pamungkas, Joko Praditi, Citra Radiana D. Antarianto Rahmawati Ridwan Reni Paramita Rini Puspitaningrum RONDANG ROEMIATI SOEGIANTO Rostika Flora Saleh, Mgs. M. Irsan Samsuridjal Djauzi Sari, Dewi H. Sarsanti, Pungguri Ayu Nega Satriotomo, Irawan Septelia I. Wanandi SEPTELIA INAWATI WANANDI SEPTELIA INAWATI WANANDI Siregar, Nurjati Siti Rahmawati Achyat Siufui Hendrawan Sri S. Ningsih, Sri S. Sri W.A. Jusman Sri Widia A.Jusman Subandrate Syarifah Dewi Tiwuk Susantiningsih Tomohiko Yamazaki Trismilah Trismilah TRISMILAH TRISMILAH Trismilah, Uly A. Nikmah, Uly A. Wahono Sumaryono Wardaya Wardaya Wawan Mulyawan Wawan Mulyawan Yefta Moenadjat Yudianita Kesuma, Yudianita Yuhernita . Zen Hafy