The Indonesian Biomedical Journal

94-125

Authors: Irma Ruslina Defi, Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Fibronectin type III domain-containing protein 5 (FNDC5), or also known as irisin, has been identified for two decades but almost completely disregarded for 10 years. It is present in skeletal muscle, heart, and brain, and in reaction to exercise can transform white adipose tissue into brown. Since then, irisin has gained a lot of attention for its potencies in treating metabolic disorders. In this review article, the potential future of irisin especially on metabolism and aging process will be discussed. CONTENT: Sedentary lifestyle is acknowledged as risk factor for type 2 diabetes mellitus, obesity, immune system issues, asthma, and neurological or heart illness. Irisin is secreted by muscle cells when exercising, produced after the proteolytic cleavage of FNDC5 protein. Irisin has positive impacts on maintaining physiological balance including reducing inflammation, keeping the bone homeostasis, as well as influencing metabolic processes and the neurological system function. Due to these many and advantageous characteristics, irisin could be a possible choice for preventing and managing disorders associated with modern society, and finding the agents to increase irisin can be beneficial. SUMMARY: Irisin offers a fresh potential basis for kinesitherapy and shows promise as a therapeutic target due to its various biological activities. Irisin pathway can be activated through dietary changes, the use of natural substances and drugs and can interact with this signalling pathway which involved peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) and uncoupling protein mRNA 1 (UCP1) to resolve obesity and its metabolic comorbidities.

135-43

Authors: Primariadewi Rustamadji, Elvan Wiyarta, Meike Pramono, Sinta Chaira Maulanisa

BACKGROUND: Neoadjuvant chemotherapy (NAC) plays a crucial role in the management of invasive breast cancer with no special type (IBC-NST), with the immune system's response to cancer heavily relying on the dynamics between tumor-infiltrating lymphocytes (TILs) and cancer cells. In this study, the differential effects of anthracycline-based NAC on stromal and intratumoral foxhead box P3 (FOXP3+) TILs expressions were specifically examined. METHODS: In this cross-sectional study, 32 IBC-NST samples were evaluated for pre- and post-NAC FOXP3+ TIL expression as well as the changes of FOXP3+ TIL expression. Comprehensive data collection regarding subjects' age, tumor size, grade, lymphovascular invasion, regional lymph node metastasis, and receptor status were conducted. Immunohistochemistry was utilized to quantify FOXP3+ TILs. The stromal, intratumoral and total FOXP3+ TILs expression were then analyzed. RESULTS: Significant reductions in FOXP3+ TIL expression post-NAC were observed, with stromal FOXP3+ TILs showing a median decrease of 3.6 units in subjetcs aged ≥50 years (p=0.013) and a median decrease of 13.2 units in subjects with tumors ≥5 cm after NAC (p=0.006). In contrast, intratumoral FOXP3+ TILs remained relatively stable, with minor changes. The total FOXP3+ TIL expression, combining stromal and intratumoral components, was significantly decreased with a median of 13.0 units decreased to 5.3 units (p<0.001). CONCLUSION: This study highlights the significant reduction in stromal FOXP3+ TIL expression after NAC treatment in IBC-NST subjects, in contrast to the relatively stable intratumoral FOXP3+ TILs. Understanding these differences may guide future therapeutic strategies and improve treatment outcomes for IBC-NST.

152-61

Authors: Sri Yani, Salim Harris, Ahmad Aulia Jusuf, Roman Goenarjo, Feda Anisah Makkiyah, Rifqi Sabita, Nurhadi Ibrahim

BACKGROUND: Stroke is one of the major causes of disability in the world. High-intensity interval training (HIIT) is known as a novel treatment to promote stroke recovery. However, the results differ in their effects on irisin, which is a regulator of brain-delivered neurotrophic factor (BDNF). Therefore, this study was conducted to investigate the effect of HIIT on BDNF and irisin levels in a rat model of ischemic stroke with middle cerebral artery occlusion (MCAO) induction on recovery motor function. METHODS: Rats were categorized into 4 groups: sham, MCAO, MCAO+moderate-intensity interval training (MIIT), and MCAO+HIIT. MCAO induction was performed to create the ischemic stroke rats model. The motor function was assessed through rotarod and footprint tests. Blood samples were obtained 6 days before MCAO and 14 days after MCAO to examine BDNF and irisin levels with enzyme-linked immunosorbent assay (ELISA). Brain tissue samples were collected 14 days after MCAO for histopathological examination of cortical tissue with hematoxylin-eosin (HE) staining. RESULTS: Rats in the MCAO+HIIT group exhibited an enhanced ability to walk on the rotarod (p=0.016). The stride-length hind paw right in the MCAO+HIIT group demonstrated a noteworthy increase in comparison to baseline value (p=0.036), and the stride-length fore paw right showed a significant increase in both the MCAO+HIIT (p=0.036) and MCAO groups (p=0.034). BDNF significantly improved in the MCAO+MIIT (p=0.043) and MCAO+HIIT groups (p=0.018). The irisin level only showed a significant enhancement in the MCAO+HIIT group (p=0.018). CONCLUSION: HIIT increased motor function, while BDNF level increased with HIIT and MIIT intervention. This preclinical research is useful for supporting the recovery of stroke patients by HIIT intervention.

172-9

Authors: Merci Monica br Pasaribu, Suzanna Immanuel, Damayanti Rusli Sjarif, Ina Susianti Timan, Safarina G. Malik, Muchtaruddin Mansyur, Ernawati Simanjuntak

BACKGROUND: Amino acids are one of the essential metabolites, especially the 20 amino acids that are preserved as the building blocks of protein. Alterations in amino acid concentrations are related to disease such as inborn error of metabolism, cancer, as well as nutritional status. Hence, it is necessary to define reference values of 20 plasma-free amino acids for Indonesian toddlers and to establish a robust measurement technique using chromatography with tandem mass spectrometry (MS). METHODS: This study was a cross-sectional preliminary study to establish reference values. The sample was prepared by mixing plasma with 20% sulfosalicylic acid. Plasma-free amino acids were measured with high-performance liquid chromatography (HPLC) non-derivatization technique using column XTerra for chromatographic separation coupled with tandem MS. Amino acids reference values were taken from 101 healthy Indonesian toddlers aged 1-3 years old. Since amino acids data were not Gaussian distributed, the lower and upper of the reference value was established from the 5th percentile and the 95th percentile, respectively. RESULTS: Analysis for 20 amino acids was validated. The accuracy ranged from 90.53-105.39% and the precision ranged from 0.06-3.80%. The limit of detection range was 1-2 nmol/mL, and the limit of quantification range was 2-4 nmol/mL. The result was linear, with R2 higher than 0.998. There was no significant difference between boys and girls for all amino acids except for glycine. CONCLUSION: HPLC with tandem MS method can be used to evaluate amino acids in clinical practice. The reference values obtained are specific for aged 1-3 years old from urban areas in Indonesia. The study suggests that for each population, the reference values for amino acids should be established.

189-98

Authors: Ermin Rachmawati, Maulidya Machdaniar, Larasati Sekar Kinasih, Djanggan Sargowo, Brillian Putra Syarifuddin, Fayza Cinta Rahmadina Putri, Mahrus Ismail

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD), characterized by inflammation and steatosis, is one of the metabolic problems that trigger serious complications. Statin is usually used for NAFLD therapy, however, statin intolerance and resistance reduce the effectiveness of NAFLD treatment. Therefore, this study was conducted to develop the phytopharmaca against NAFLD by elucidating the anti-inflammatory and steatosis suppression of vegetable spices fermentation extract (VSFE) in rabbits induced with high-fat diet (HFD). METHODS: Twenty New Zealand White rabbits were determined into 4 groups; which were groups receiving standard diet (SD), HFD, HFD and 100 mg/kgBW VSFE, as well as HFD and 200 mg/kgBW VSFE. mRNA expressions of tumor necrosis factor alpha (TNFα) and CCAAT/enhancer-binding protein alpha (C/EBPα) were evaluated. The TNFα concentration was quantified after protein and RNA isolation. Inflammation and steatosis were observed in liver hematoxylin-eosin-stained preparate using microscope. RESULTS: Macrophage cells and hepatic steatosis showed significant decrease in a drug dependent manner. An effective dose of 200 mg/kgBW decreased C/EBPα and TNFα expression to 0.45±0.32 and 0.72±0.29 (p=0.013 and p=0.002, compared to the HFD group), respectively. TNFα inflammatory cytokine concentrations also responded to the administration of 100 mg/kgBW and 200 mg/kgBW doses, with the value of 31.72±10.40 mg/dL and 48.35±7.15 mg/dL (p=0.009 and p=0.002, compared to the HFD group), respectively. CONCLUSION: VSFE might prevent NAFLD by inhibit steatosis and inflammation through suppression of C/EBPα expression and TNFα expression.

1-22

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: World Health Organization has reported four million people die every year due to obesity comorbidity, and the prevalence of obesity is keep increasing, especially after COVID-19. Obesity has been defined as a chronic disease involving adipose tissue dysfunction which leads to metabolic diseases and psychosocial consequences. The review article will highlight some recent researches regarding the new conceptual framework that integrates both metabolic drives, as well as to summarize the numerous discussions about the current understanding of hypothalamic control of food intake and energy homeostasis. CONTENT: Obesity apparently is not simply regulated only by food and exercise. Hypothalamus takes part in controlling energy intake and expenditure via appetite regulation. Hedonic control in cortical and subcortical brain areas process cognitive, reward, information, and executive function. Managing metabolic adaptation, browning the white adipose tissue, and preserving lean mass can be another strategy to safely manage obesity. SUMMARY: Obesity need to be managed in a multimodal strategy including neurophysiology and physiology approach, together with environment support. Thus, a weight regain can be prevented. Commitment from both scientific and regulation point of view can shed a light to eradicate obesity.

31-9

Authors: Edy Fachrial, Juliana Lina, Harmileni, Sari Anggraini, Widya Yanti Sihotang

BACKGROUND: Type 2 diabetes mellitus (T2DM) cases tend to rise throughout the year in all parts of the world. The α-glucosidase inhibitors are frequently used to both prevent and treat T2DM. This α-glucosidase inhibitor activity is seen in some probiotic species, and a certain strain of Pediococcus acidilactici exhibits promising characteristics as an α-glucosidase inhibitor. This study was conducted to assess the hypoglycemic activity and safety of P. acidilactici strain DNH16 (PADNH16) in experimental rats with T2DM induced by streptozotocin. METHODS: The experiment was conducted in vivo using Wistar rats. Acarbose was employed as a positive control, and Lactobacillus casei was used as a comparative. For 24 days, post-prandial blood glucose levels were assessed every 3 days, followed by serum biochemical levels measurement of liver, kidney, and lipid profiles. The pancreas was histopathologically examined utilizing the Hematoxyline-Eosin staining procedure. RESULTS: Administration of PADNH16 to T2DM rats lowered post-prandial blood glucose levels and gave hypoglycemic benefits comparable to acarbose and commercial probiotics. PADNH16 dosing did not affect serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), urea, or creatinine levels, showing that PADNH16 was not hazardous to liver or kidney function. The lipid profile assessment revealed that the values, including high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides, were comparable to the control group. The pancreatic histopathology revealed that injection of PADNH16 caused no alterations to pancreatic β cells. CONCLUSION: P. acidilactici isolate DNH16 has a hypoglycemic effect on T2DM rats, but does not affect liver function enzymes, kidneys, lipid profiles and does not provide significant changes in pancreatic β cells.

48-55

Authors: Muhammad Ridwan, Herlina Dimiati, Maimun Syukri, Ronny Lesmana, Lia Meuthia Zaini

BACKGROUND: Sodium glucose co-transporter-2 inhibitor (SGLT2-i), a new oral antidiabetic drug, has been recommended for its morbidity and mortality benefits in patients with heart failure. This study aimed to determine the effect of acute SGLT2-i therapy on the relative ratio expression of microRNA-21 (miR-21), transforming growth factor-β1 (TGF-β1), and matrix metalloproteinase-2 (MMP-2) in the process of cardiac fibrosis in the hyperglycemia Wistar rat models compared to biguanide (metformin) therapy. METHODS: We used Streptozotocin (STZ) to induce hyperglycemia in Wistar rats (n=31), randomly divided into four groups: negative control (NC, n=4), positive control (PC, n=10), hyperglycemia plus metformin (M, n=8), and hyperglycemia plus empagliflozin (E, n=9). After seven weeks, the rats were sacrificed and the heart tissue was taken for microRNA and messenger RNA (mRNA) extraction, followed by reverse transcription quantitative real time polymerase chain reaction (RT-qPCR) examination. The data was analyzed using One-way ANOVA. RESULTS: Results showed a decreasing trend in the gene expression relative ratio of miR-21 (1.0 vs. 1.9; p=0.079) and TGF-β1 (0.9 vs. 3.2; p=0.145), but a significant increase in MMP-2 gene expression (1.3 vs. 0.7; p=0.002) in the SGLT2-i (empagliflozin) vs. biguanide (metformin) groups. CONCLUSION: Empagliflozin administration may play a significant role in preventing the occurrence of cardiac fibrosis in hyperglycemia.

66-71

Authors: Ryco Giftyan Ardika, Bernardus Parish Budiono, Nyoman Suci Widiastiti, Nani Maharani, Neni Susilaningsih, Ferry Sandra

BACKGROUND: Microcirculation and cellular disturbances caused by sepsis might trigger significant intestinal damage. Andrographis paniculata extract decreases inflammatory intestinal epithelial cells with its role as an antiparasitic and anti-inflammatory agent. However, A. paniculata extract’s effect on sepsis have not been commonly studied, especially in the intestinal tissues. Therefore, this study was conducted to determine A. paniculata leaves extract (APLE) effect in sepsis-induced intestinal tissues of rats by examining the expression of inflammatory cytokines involved in sepsis, namely tumor necrosis factor (TNF)-α and Caspase-3. METHODS: Rats were divided into five groups; two groups received no pretreatment and the other three groups received 200, 400, and 500 mg/kg BW/day APLE, respectively. Three pretreated groups and one group with no pretreatment were then injected with 1 mg/200 g BW lipopolysaccharides (LPS) intraperitoneally to create septic rat models. Three days after the LPS-induction, rats were euthanized and the expression of TNF-α and Caspase-3 were assessed based on the immunohistochemical staining of rats’ intestinal tissues. RESULTS: Compared with NaCl (sham), LPS significantly (p<0.001) induced TNF-α expression from 6.60±1.36 to 25.37±1.74. Pretreatment of 200, 400, and 500 mg/kg BW/day APLE could significantly (p<0.001) inhibit the LPS-induced TNF-α expression (18.82±1.36, 11.45±1.18, and 6.89±1.90, respectively). Similar with TNF-α, compared with NaCl (sham), LPS significantly (p<0.001) induced Caspase-3 expression from 6.92±1.66 to 23.59±2.25. Pretreatment of 200, 400, and 500 mg/kg BW/day APLE could significantly (p<0.001) inhibit the LPS-induced Caspase-3 expression (17.47±1.68, 12.99±1.51, and 5.59±1.51, respectively). CONCLUSION: The pretreatment of APLE could inhibit the LPS-induced TNF-α and Caspase-3 expression, therefore APLE could be suggested as a potential sepsis-preventing agent.

79-87

Authors: Ujang Saeful Hikmat, Ani Retno Prijanti, Heri Wibowo, Indriyanti Rafi Sukmawati, Dicky Levenus Tahapary

BACKGROUND: Type 2 Diabetes (T2D) impairs the innate immune system including monocytes. Monocytes are divided into two subgroups depending on the expression of cluster of differentiation (CD)14 and CD16 receptors, namely CD14+CD16- and CD14+CD16+. CD14+CD16+ are proinflammatory monocytes and develop into M1 type macrophages, which contribute to foam cell production, a risk factor for cardiovascular disease (CVD). Therefore, it is important to determine the influence of T2D conditions on changes in monocyte subsets and whether these changes correlate with CVD risk markers. METHODS: Peripheral blood mononuclear cell (PBMC) was obtained from 10 T2D subjects and 10 healthy donors. Subsequently, PBMC was incubated for 24 hours with and without 10 mL lipopolysaccharide. Flow cytometry was used to evaluate CD14 and CD16 expression, while multiplex immunoassays were applied to measure interleukin (IL)-1b and IL-10 concentrations in supernatants. RESULTS: In T2D, the percentage of CD14+CD16+ monocytes increased (p=0.07), and an increase in CD14+CD16+ monocytes more than 6.8% was linked with CVD risk markers (r=10.146, p=0.002). Meanwhile, inflammatory mediators released by monocytes shown an increase in IL-1b (p=0.041) but not in IL-10 (p=0.082) in T2D subjects. Fasting blood glucose levels were also found to be substantially linked with an increase in CD14+CD16+ monocytes (r=0.530, p=0.016). CONCLUSION: T2D patients had a higher percentage of CD14+CD16+ monocytes and IL-1b levels than healthy donors. An increase in CD14+CD16+ monocytes above 6.8% associated with CVD risk markers in T2D patients.

351-7

Authors: Muhammad Agus Aulia, Keri Lestari, Ahmad Faried, Cynthia Retna Sartika, Andi Wijaya, Achmad Adam, Hanna Gunawan, Rima Haifa, Nabilla Farah Naura, Billy Yosua Costantin Pongajow

BACKGROUND: Stroke is one of many leading causes of death and disability worldwide. Despite decades of research, treatment for stroke recovery is still inadequate. Recently, mesenchymal stem cell (MSC) therapy is considered as one of the potential treatments for stroke due to their capabilities to repair or regenerate damaged tissues, as well as immunomodulators in inflammatory conditions. Many pre-clinical and clinical trials have shown MSC therapy feasibility, safety, and efficacy in treating stroke. However, evidence regarding the optimal treatment plan and factors that can improve stem cell functional outcomes in treating stroke needs to be further explored. Therefore, a systematic review was conducted. METHODS: Recent literatures from 2015-2022 regarding stem cell therapies, specifically MSC on ischemic stroke, were collected from two reliable databases: PubMed and PubMed Central. Collected literatures were properly applied to inclusion-exclusion criteria and appraised critically. Keyword strategies on databases were employed, including both medical subject headings (MeSH). RESULTS: Five literatures from 726 were identified and used for systematic review. All animal model in the literatures were prepared to have middle cerebral artery occlusion. All studies indicated that MSC therapy is a safe and reliable procedure despite the variety of transplantation routes. No report of toxicity, rejection reaction, nor infection on MSC treated groups. CONCLUSION: Stem cell sources, dosages, and delivery routes could be resourceful for future translational studies to ensure the safety and efficacy of MSC therapy for ischemic stroke.

366-74

Authors: Irma Rahayu Latarissa, Melisa Intan Barliana, Anna Meiliana, Ida Paulina Sormin, Erizal Sugiono, Cissy Bana Kartasasmita, Irmansyah Irmansyah, Keri Lestari

BACKGROUND: Before WHO revoked the emergency use authorization for Chloroquine (CQ) and Hydroxychloroquine (HCQ) because of their side effects, it was suggested to use these two drugs for COVID-19 therapy. In addition, another derivate of quinine, namely Quinine Sulfate (QS), showed good in silico and in vitro antiviral activity against SARS-CoV-2. Prior the WHO revocation, this study was conducted to evaluate the efficacy of QS in mild-to-moderate COVID-19 patients. METHODS: This was an adaptive, controlled, multicenter, randomized, double-blind clinical trial involving mild-to-moderate COVID-19 patients in Indonesia. The participants were divided into 2 groups: the control group (standard COVID-19 treatment + placebo) and the treatment group (standard COVID-19 treatment + QS). The primary outcome was the efficacy of QS based on clinical status using a 7-point ordinal scale. The secondary outcomes were the efficacy of QS in terms of the incidence and duration of oxygen supplementation, incidence of mechanical ventilation, and length of stay. RESULTS: No significant difference in the efficacy parameters studied was found between the control group and the treatment group. The difference in the mean oxygen saturation was also measured and the results showed a significant difference where the treatment group had higher mean oxygen saturation than the control group (p=0.001). CONCLUSION: Although not significant, the treatment group showed better therapy outcomes compared to the control group.

383-90

Authors: Rina Triana, Indriyanti Rafi Sukmawati, Mas Rizky Anggun Adipurna Syamsunarno, Keri Lestari

BACKGROUND: Type 2 diabetes mellitus (T2DM) is the most common metabolic disorder associated with oxidative stress and chronic inflammation. Understanding the regulatory mechanisms related to the role of nuclear factor erythroid 2 (Nrf2), a master regulator of cellular antioxidant defenses, in individuals with T2DM, is essential. Therefore, in-depth investigation regarding the associations between Nrf2 levels, and metabolic parameters, such as waist circumference, fasting glucose levels, HbA1C, and serum inflammatory markers expressions is needed to elucidate the mechanisms driving T2DM and its metabolic disturbances. METHODS: This was a cross-sectional study including 90 T2DM and 25 healthy subjects. Nrf2 level; serum inflammatory markers levels, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, interferon (IFN)-α2, IFN-g, monocyte chemoattractant protein (MCP)-1, IL-10, IL-12p70, IL-17A, IL-18 and IL-23; as well as waist circumference; fasting glucose levels; and HbA1C were measured. Nrf2 concentrations in peripheral blood mononuclear cells were determined using enzyme-linked immunosorbent assay (ELISA), while serum inflammatory markers concentrations were quantified with flowcytometry. RESULTS: Nrf2, TNF-α, and IL-17A levels were significantly higher in the T2DM group, while IL-1β, IFN-α 2, IL-10, IL-12p70, and IL-23 levels were elevated in healthy controls Nrf2 exhibited positive correlations with waist circumference, fasting glucose, HbA1C, and TNF-α. Conversely, an inverse correlation of Nrf2 was observed with IL-1β, IL-12p70 and IL-17A. CONCLUSION: Correlations between Nrf2 and metabolic clinical parameters suggest its role in regulating glucose metabolism and adiposity. Elevated Nrf2 levels observed in T2DM patients may present novel therapeutic avenues for enhancing endogenous antioxidant defenses.

400-10

Authors: Nur Irawati, Heri Wibowo, Ani Retno Prijanti, Rona Kartika, Dicky Levenus Tahapary

BACKGROUND: Cluster of differentiation (CD)36 and adenosine triphosphate-binding cassette transporter A1 (ABCA1) are 2 macrophages-expressed receptors that promote cholesterol uptake and efflux, in which their imbalance might be associated with the foam cell formation risk. Type 2 diabetes mellitus (T2DM) has been correlated with the increase of this plaque formation. Therefore, it is necessary to determine whether expression of CD36 and ABCA1 in macrophages are correlated with coronary artery disease (CAD) risk markers in T2DM cases. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from 13 diabetic patients and 11 healthy donors. Then, the PBMC-derived macrophages were cultured with supplement of oxidized low-density lipoprotein (ox-LDL) or lipopolysaccharide (LPS). Expression of CD36 and ABCA1 was measured using flowcytometry, meanwhile the supernatant concentration of interleukin (IL)-1b and IL-10 was measured by multiplex immunoassay. RESULTS: T2DM subjects more likely to have low proportion of CD36-ABCA+ macrophages compared to healthy donors (p=0.041) and it had negative correlation with glucose homeostasis and insulin resistance markers, including fasting blood glucose (FBG, r=-0.408, p=0.048), glycated hemoglobin (HbA1c, r=-0.380, p=0.049), triglyceride glucose index (r=-0,518, p=0.009), and high-sensitivity C-reactive protein (hs-CRP, r=-0.556, p=0.005). Moreover, it also had a negative correlation with atherogenic markers such as triglyceride (r=-0.417, p=0.043), triglyceride/HDL, and LDL/HDL, but had positive correlation with HDL (r=0.540, p=0.007). Most of T2DM subjects had high IL-1β/IL-10 ratio after ox-LDL and LPS stimulation (p=0.02 and p=0.05, respectively). CONCLUSION: Reduced proportion of CD36-ABCA1+ macrophages followed with high IL-1β/IL-10 can be a marker of CAD in T2DM.

420-8

Authors: Lina Hadi, Albert Manggading Hutapea, Florenly Florenly

BACKGROUND: Orthodontic tooth movement (OTM) may increase the risk of treatment-related complications for some diabetes mellitus (DM) patients. Hydrogen-rich water (HRW) has been demonstrated in many studies to reduce oxidative stress and cell damage. This study aimed to examine the levels of interleukin (IL)-1β, blood glucose level, body weight, tooth displacement, and population of osteoclasts and osteoblasts in diabetic rats with OTM. METHODS: Thirty rats (Rattus novergicus) were divided into 6 groups: OTM, HRW, DM, DM+OTM, DM+HRW, and DM+OTM+HRW. DM, DM+OTM, DM+HRW, and DM+OTM+HRW groups were induced with streptozotocin (STZ) after 8 weeks of high-fat diet (HFD) and continued being fed HFD for 4 weeks. OTM, DM+OTM, and DM+OTM+HRW groups were placed on an orthodontic device for the orthodontic treatment. HRW, DM+HRW, and DM+OTM+HRW groups were administered HRW via oral gavage 3 times a day for 4 weeks. At the end of the study, all rats were euthanized and blood samples were collected for IL-1β measurement using enzyme-linked immunosorbent assay (ELISA) kits. Meanwhile, the rats’ maxilla was taken to measure tooth movement, and the number of osteoclast and osteoblast were counted. RESULTS: The highest increase of IL-1β was in the DM+OTM group (140.07±5.14 pg/mL) and the lowest was in the HRW group (92.80±2.89 pg/mL). The average number of osteoblasts were higher in tension sites, while osteoclasts were higher in pressure sites. CONCLUSION: Consumption of HRW in STZ-induced diabetic rats with OTM can reduce IL-1β levels, reduce tooth mobility, and promote bone remodeling.

269-77

Authors: Albertus Budi Sulistya, Rima Haifa, Geofanny Facicilia, Kristin Talia Marbun, Marsya Nilam Kirana, Yussy Afriani Dewi, Cynthia Retna Sartika, Andi Wijaya, Keri Lestari Dandan

BACKGROUND: Dendritic cell (DC)-based cancer therapy is a promising adjuvant therapy for nasopharyngeal cancer (NPC) after chemoradiation. Owing to low immunity after chemoradiation, DC therapy activates immune responses. Moreover, DC-based cancer therapy can decrease tumor progression, prolong lifespan, and increase the quality of life of patients. Various studies regarding the use of DC therapy for NPC have been reported, however there are limited reviews on the implementation and foundation of DC immunotherapy to expand this technology. METHODS: A literature search was performed on EMBASE, ScienceDirect, PubMed (MEDLINE), and Cochrane Library, with the term dendritic cells therapy for nasopharyngeal cancer, dendritic cell immunotherapy in nasopharyngeal cancer patients, and DC therapy in NPC, as the search keywords. RESULTS: A total of 199 literatures were reviewed, and four clinical trials were identified as relevant for this review. DC vaccines can be processed with various maturation and activation processes. Selected literatures reported antigens used when incubating the DC are latent membrane protein (LMP) 1, LMP2, and Epstein–Barr virus nuclear antigen 1 (EBNA1). Although DC therapy was produced from different pathways, it has been reported that there are increases of cluster of differentiation (CD)8+ T cells, CD4+ T cells, and the progression free survival (PFS) rate in DC immunotherapy patients than the radiochemotherapy patients. CONCLUSION: It can be concluded that DC could be used as an adjuvant therapy alongside the standard therapy of NPC, which prolongs NPC patient survival.

288-96

Authors: Septelia Inawati Wanandi, Resda Akhra Syahrani, Ayu Suraduhita, Elvira Yunita, Melva Louisa

BACKGROUND: Breast cancer stem cells (BCSCs) have been identified as playing a crucial role in therapeutic resistance. This resistance can be attributed to the anti-apoptotic protein survivin and the antioxidant MnSOD high expression. To overcome the resistance to doxorubicin (DOX), this study proposed the utilization of andrographolide (ANDRO), the primary bioactive compound in Andrographis paniculata leaves. The objective was to examine the role of andrographolide in regulating survivin, caspase-9, and caspase-3 gene expressions to reverse doxorubicin resistance in human BCSCs. METHODS: BCSCs were exposed to 0.1 µM DOX every two days or 50 µM rotenone (ROT) for 6 hours, subsequently supplemented with 0.3 mM ANDRO. Superoxide and peroxide levels were measured using DHE and DCFH-DA assay. The MnSOD, survivin, caspase-9, and caspase-3 mRNA expression levels were analyzed using qRT-PCR. Protein expressions were evaluated using Western blotting assay. MnSOD activity was determined using xanthine oxidase inhibition assay. The apoptotic cells were determined using Annexin-V/PI staining. RESULTS: This study indicated that the cytotoxic mechanisms of DOX, similar to ROT, in BCSCs were attributed to oxidative stress, as evidenced by an elevation in superoxide rather than peroxide levels, accompanied by a decrease in MnSOD activity. This study also highlighted that ANDRO reversed DOX resistance in BCSCs subjected to repeated DOX treatment by downregulating survivin and upregulating caspase-9 and caspase-3 mRNA expressions, thereby activating the intrinsic apoptotic pathway. CONCLUSION: This study provides insights into the role of ANDRO in modulating the expression of apoptotic genes, such as survivin, caspase-9, and caspase-3, to overcome DOX resistance in BCSCs.

304-10

Authors: Nurul Hidayah, Ika Yustisia, Rosdiana Natzir, Lia Hafiyani, Ilhamuddin Ilhamuddin, Hijral Aswad, Marhaen Hardjo

BACKGROUND: Insufficient glucose levels in colorectal cancer (CRC) patients leads to a condition where fructose might become an alternative source for cells proliferation, but the role of fructose or fructose-glucose combinations in development of CRC has not been elucidated well. In this study, the effect of fructose-glucose variations on viability, migration, and glucose transporter (GLUT)5, GLUT7, GLUT11 mRNA expressions in WiDr CRC cell line were examined. METHODS: Cells were treated with varying ratios of fructose-glucose (F100%; F75%:G25%; F50%:G50%; F25%:G75%; G100%; F: Fructose, G: Glucose). Untreated cells (F0:G0) were used as cell control. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used for cell viability test, scratch assay was used to examine the cell migration, and quantitative polymerase chain reaction (qPCR) was performed to examine mRNA expressions. Data were analyzed using one-way analysis of variance (ANOVA) and followed with Tukey's post-hoc test, with p<0.05 consideres as significant. RESULTS: Fructose-glucose combinations and glucose 100% significantly increased the cell viability compared to control (p<0.05). All treatment groups showed a significant increase in cell migration compared to control (p=0.000). Only GLUT7 and GLUT11 expressions in the G100% group were significantly different compared to the control (p=0.000). GLUT7 and GLUT11 expressions were also significantly different in F100% and F50%:G50% treatments compared to G100% (p=0.000). CONCLUSION: Taken together, fructose might play important role in cell migration. However, in cell viability, combination with glucose could increase fructose's effect. Fructose might not affect the mRNA expressions of GLUT5, GLUT7 and GLUT11.

318-27

Authors: Desty Restia Rahmawati, Arief Nurrochmad, Riris Istighfari Jenie, Edy Meiyanto

BACKGROUND: The anti-cancer properties of pentagamavunon-1 (PGV-1) and curcumin have been documented. This study aimed to evaluate the cytotoxic effect of this combination on breast cancer cell growth using 4T1 and T47D cells, representing triple-negative breast cancer (TNBC) and non-TNBC, respectively. METHODS: Cytotoxic assay was evaluated using MTT reagent for single and combination treatment of PGV-1 and curcumin in 4T1 and T47D cells. Cell cycle analysis was examined through flowcytometry with propidium iodide dye. May Grünwald-Giemsa staining was also performed to analyze the mitotic catastrophe RESULTS: PGV-1 and curcumin alone had significant cytotoxic effects against two breast cancer cell lines, with IC50 values of 4 μM and 40 μM for 4T1 and 2 μM and 20 μM for T47D, respectively. Both compounds showed high selectivity for the 4T1 and T47D cells (selectivity index >3). In addition, when PGV-1 and curcumin were combined, a synergistic effect was observed in both cell types with a combination index of <0.7. This combination results in cell cycle arrest in the G2/M phase, increased cell accumulation in the sub-G1 phase, and a synergistic increase in mitotic catastrophe. CONCLUSION: Combined intervention of PGV-1 and curcumin on TNBC and non-TNBC breast cancer cells substantially augments cell cycle arrest in the G2/M and sub-G1 phases, coupled with the occurrence of mitotic catastrophe. In summary, the results suggest that PGV-1 coupled with curcumin holds promise as an effective approach to addressing breast cancer and warrants further investigation.

336-40

Authors: Nur Zam Zam, Harun Iskandar, Nur Ahmad Tabri, Arif Santoso, Nurjannah Lihawa, Harry Akza Putrawan, Ferry Sandra

BACKGROUND: Emergence of drug resistance due to epidermal growth factor receptor (EGFR) Exon 20 T790M poses a challenge in the effective management of non-small cell lung carcinoma (NSCLC). Significant breakthrough in the management of NSCLC with a specific genetic alteration causes substantial condition improvement in patients whose cancer progressed after first-generation tyrosine kinase inhibitor treatment and who developed tumors with EGFR Exon 20 T790M mutation. The present study analyzed a cohort of NSCLC patients and investigated the incidence of the EGFR Exon 20 T790M status with Osimertinib therapy, along with its impact on survival rates. METHODS: This was a retrospective cohort study on 22 NSCLC subjects who were genetically examined for EGFR status from plasmic cell free total nucleic acid. Subjects with EGFR Exon 20 T790M mutation were treated with/without Osimertinib. Demographic and clinical data were descriptively summarized, and the differences of each variable and correlation between survival rate and EGFR Exon 20 T790M were analyzed. RESULTS: Subjects with (n=13) and without (n=9) EGFR Exon 20 T790M had survival rates of 10.77±2.45 and 4.78±1.48, respectively (p=0.000). Based on 1-year survival status of subjects with EGFR Exon 20 T790M, there were 3 Osimertinib-treated survivors and 2 Osimertinib-treated non-survivors. Eight subjects with EGFR Exon 20 T790M and without Osimertinib treatment did not survive (p=0.001). CONCLUSION: Since the treatment of Osimertinib demonstrated a noteworthy survival rate among NSCLC subjects with EGFR Exon 20 T790M, thus Osimertinib could be suggested as a potential targeted therapy for NSCLC subjects with EGFR Exon 20 T790M.

296-302

Authors: Maki Yoshikawa, Masatoshi Oshita, Tomomi Gotoh, Yukina Tawa, Katsuaki Dan

BACKGROUND: Stem cell culture medium is garnering attention because it comprises growth factors and exosomes secreted by stem cells. In this study, stem cell culture medium was subjected to electric field stimulation using the cells alive system (CAS) to determine the antiaging effects of this medium on dermal fibroblasts. METHODS: The medium was added to the monolayer cultures of human fibroblasts with or without 100 μg/mL advanced glycation end products (AGE), and the cells were incubated for 4 hours. Total RNA was extracted, and mRNA levels of several AGE receptors and heat shock proteins were measured using quantitative reverse transcriptase-polymerase chain rection (qRT-PCR). RESULTS: Similar amounts of exosomes were extracted from the supernatants of stem cells cultured with (CAS+) and without (CAS−) electric field stimulation. The results demonstrated that CAS+ had a higher oxygen radical absorbance capacity than CAS−, indicating its high antioxidant activity. Furthermore, the expression of domain-containing link scavenger receptor-1 mRNA, which is involved in the degradation of cell surface receptors for AGE under glycation stress, significantly increased. The electric field stimulation also increased the expression of several heat shock proteins, which play important roles in cellular responses generated against glycation stress. The electric field stimulation of the stem cell culture medium effectively promoted the endocytosis and degradation of AGE, thereby exerting antioxidant effects and inducing glycation stress. CONCLUSION: Electric field stimulation of stem cell culture medium enhanced the expression of Hsp and its antioxidant activity in fibroblasts.

310-5

Authors: Faisal Faisal, Nur Rochmah, Muhammad Faizi, Novina Novina, Erni Nuraeni

BACKGROUND: Type 1 diabetes (T1D) mostly occurs due to the destruction of pancreatic beta cells due to autoimmune processes. Diagnosis of T1D can be established by examining the c-peptide levels and the markers of pancreatic autoantibodies, including glutamic acid decarboxylase 65 autoantibodies (GAD-65). However, the association between c-peptide and anti-GAD-65 toward patients’ clinical manifestations needs to be further explored. Hence, the aim of current study was to identify the association of anti-GAD65 with c-peptide and clinical characteristics in children with T1D. METHODS: Case-control study involving 47 T1D children (T1D group) and 41 healthy children (control group) younger than 18 years old was conducted. Secondary data regarding subjects’ demographic characteristics and medical history were collected from subjects, and serum blood was drawn from each subject for the anti-GAD65 and c-peptide measurement. Anti-GAD65 and c-peptide levels were measured using an Enzyme-Linked Immunosorbent Assay (ELISA) methods. RESULTS: Anti-GAD65 antibody was detected in 78.7% T1D group, while only 2.43% were detected in control group subject (p=0.0000). The c-peptide level of T1D group was 0.07±0.19 nmol/L and control group was 1.5±0.77 nmol/L (p=0.0000). The total daily dose of insulin in subjects with positive anti-GAD65 was greater than in the negative anti-GAD65 (p=0.012). The sensitivity and specificity of the anti-GAD65 were 85.4% was 66.7%, respectively. CONCLUSION: The results of this study show that anti-GAD65 was associated with total daily dose of insulin requirement in children with T1D.

322-32

Authors: Dinda Aliffia, Dinda Ayu Ramadhani, Widya Wasityastuti, Dewi Ratih Tirto Sari, Ulayatul Kustiati, Hevi Wihadmadyatami, Dwi Liliek Kusindarta

BACKGROUND: Neurodegenerative diseases are characterized by the loss of neuronal function in the nervous system. In recent years, more than 45 million people worldwide have suffered from progressive loss of memory and cognitive functions caused by Alzheimer’s disease. Ocimum sanctum is one of the medicinal plants known to have neuroprotective abilities. This study was conducted to elucidate the anti-apoptotic effects of ethanolic extract of O. sanctum (EEOS) on PC12 and SH-SY5Y cells as well as interaction between main compounds of EEOS and β-amyloid (Aβ) peptide through in silico molecular docking. METHODS: The viability of TMT-induced PC12 and SH-SY5Y cells was assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and acridine orange/propidium iodide staining. Cell proliferation rate was measured with cell counting kit-8 (CCK-8) assay. Nuclear fragmentation was observed with Hoechst 33342 staining. Caspase -3 and -9 expressions were measured using enzyme-linked immunosorbent assay. Interactions between main compounds of EEOS and Aβ were visualized with in silico molecular docking. RESULTS: EEOS had the potential effect of maintaining cell viability, preventing the cell’s morphological changes, and inhibiting apoptosis via the caspase pathway in PC12 and SH-SY5Y cells. Meanwhile, flavonoid K, phenol, eugenol could interact with the active site of Aβ through hydrogen-bonding and hydrophobic interactions. CONCLUSION: EEOS could prevent neuronal cell apoptosis via downregulation of caspase-3 and -9. Main compounds of EEOS could interact with the active site of Aβ, and thereby might inhibit Aβ oligomerization. Thus, EEOS and its main compounds could be potential as neuroprotective agents for preventing neurodegenerative diseases.

339-46

Authors: Yudy Tjahjono, Caroline Caroline, Jusak Nugraha, Kuncoro Foe, Srikanth Karnati, Süleyman Ergün, Nico Jafet, Oryza Chrisantia, I Made Andika Bara Kusuma, Hendy Wijaya, Wuryanto Hadinugroho, Dwi Aris Agung Nugrahaningsih, Dwi Liliek Kusindarta, Yufita Ratnasari Wilianto, Senny Yesery Esar, Hevi Wihadmadyatami

BACKGROUND: Lipopolysaccharide (LPS) has been reported to increase CD4+ regulatory T-cell (CD4+ Treg) populations. Acetylsalicylic acid (ASA) has been reported to have immunomodulatory activity, but it may induce chronic gastric ulceration. Another salicylic acid-bearing compound, 2-(3-(chloromethyl)benzoyloxy)benzoic acid (3-CH2Cl), has been reported to have less gastric mucosal damage. However, the effect of 3-CH2Cl on CD4+ Tregs in LPS-induced mice is still unknown. Therefore, the present study was conducted to investigate the immunomodulatory effect of 3-CH2Cl on CD4+ T-cell and CD4+ Treg populations as well as FoxP3 expression in LPS-induced mice. METHODS: Synthesis of 3-CH2Cl was performed by mixing salicylic acid and chloromethylbenzoylchloride with the catalyzation of pyridine, acetone and heat. The 3-CH2Cl tablets were prepared using direct compression method. After intraperitoneal injection of 1 mg/kg BW LPS to mice, 60 mg/kg BW ASA or 60 mg/kg BW 3-CH2Cl was given orally for 3 days. The splenocyte was obtained through splenectomy and collagenase digestion. The population of CD4+ T-cells and CD4+ Tregs, as well as the splenic FoxP3 expression were determined using flow cytometry technique. RESULTS: CD4+ T-cell populations in mice treated with LPS and 3-CH2Cl or ASA were lower than those treated with LPS merely. Meanwhile, CD4+ Treg populations and FoxP3 expression levels in mice treated with LPS and 3-CH2Cl or ASA were higher than those treated with LPS merely. CONCLUSION: Since 3-CH2Cl could decrease CD4+ T-cell population and increase CD4+ Treg population mediated by the increase of FoxP3 expression in LPS-induced inflammation, it may act as a potential therapeutic drug to reduce inflammatory conditions.

354-61

Authors: Bantar Suntoko, Suharyo Hadisaputro, Handono Kalim, Suyanto Hadi, Ika Vemilia Warlisti

BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune condition characterized by persistent, chronic inflammation that damages organ tissue. One of the symptoms that is often found in SLE is cognitive dysfunction. Hydroxychloroquine is recommended for the treatment of all levels of SLE. This study was conducted to prove the influence of hydroxychloroquine on improving cognitive function and inflammatory biomarkers compared to standard therapy. METHODS: The study adopted randomized controlled trial (RCT) in SLE patients with cognitive dysfunction who met the inclusion criteria. The treatment group consisted of 26 subjects who received hydroxychloroquine 200 mg/day for 8 weeks and standard therapy, while the control group consisted of 29 subjects who were given standard therapy only. Examination of Montreal Cognitive Assessment (MoCA)-INA, interleukin (IL)-6, IL-4, interferon (IFN)-α, and C-reactive protein (CRP) scores was carried out at the beginning and the end of the study. The unpaired variables were examined with independent T-test or the Mann-Whitney test, while the paired variables were examined with paired T-test or Wilcoxon signed rank test. The Spearman correlation test was used to measure correlation between variables. RESULTS: A total of 55 subjects participated and completed the study. The result showed a significant relationship between hydroxychloroquine and decreasing levels of IL-6 and IL-4 (p<0.05). Meanwhile, there was no significant effect on the increase in cognitive function and decrease in IFN-α and CRP (p>0.05) in both groups. CONCLUSION: Hydroxychloroquine decreases the levels of IL-6 and IL-4, but has no effect on cognitive function, levels of IFN-α and CRP.