The Indonesian Biomedical Journal

114-39

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: A lot of contemporary cancer research has concentrated on genetic influence. However, cancer also involves biochemical changes, such as metabolic adaptation to support the aberrant cell proliferation. CONTENT: The fast cell proliferation in cancer cells enforce a metabolic re-arrangement to promote their long-term survival. The increased glucose uptake and fermentation of glucose to lactate are common features of this altered metabolism known as “the Warburg effect”. These metabolic pathways regulation enable cancer cells to produce adenosine triphosphate (ATP) in an efficient way. Epigenetic and metabolic changes also both affect molecular rewiring in cancer cells and promote cancer development and progression. SUMMARY: Metabolic rewiring and epigenetic remodeling establishing a direct link between metabolism and nuclear transcription to promote the survival of tumor cells. A further understanding of how metabolic remodeling can result in epigenetic changes in tumors, affecting cancer cell differentiation, proliferation, and/or apoptosis, will lead to a new strategy for cancer therapy.

147-54

Authors: Erna Susanti, Endang Susilowati

BACKGROUND: Signaling pathways contributing to cholesterol efflux and inhibitory inflammation in atherosclerosis that has not been explored is the liver X receptor (LXR). Catechin as LXR agonist influences the expression of Niemann-pick C1 like 1 (NPC1L1) protein transporter that triggers the inhibition of cholesterol absorption. This study aimed to examine the effect of Catechins on the expression of intestinal transporters: NPC1L1, ATP-binding cassete-proteins G5 (ABCG5) and G8 (ABCG8). METHODS: Twenty-five experimental animals were divided into five treatment groups, with 5 rats in each group. The groups were normal diet rats (group 1), high fat diets-induced rats (group 2), high fat diets-induced rats treated with 30 mg/kg body weight (BW) of Catechins (group 3), high fat diets-induced rats treated with 60 mg/kg BW of Catechins (group 4), and high fat diets-induced rats treated with 120 mg/kg BW of Catechins (group 5). After one-month, all rats were sacrificed, blood and intestine were collected. Lipid profile were determined enzymatically, mRNA levels were determined by reversetranscriptase polymerase chain reaction (RT-PCR), while the expression of protein transporter were determined by immunohistochemistry. RESULTS: Catechins treatment decreased the expression of NPC1L1, but increased the expression of ABCG5 and ABG8. CONCLUSION: Catechins can be developed as a candidate for NPC1L1 inhibitor to mediate the inhibiting absorption of intestinal cholesterol, therefore increasing the inhibitory effect of atherogenesis.

163-9

Authors: Astuti Giantini, Ina Susianti Timan, Erlin Listiyaningsih, Rahajuningsih Dharma, Rianto Setiabudy, Idrus Alwi, Lia Gardenia Partakusuma, Nuri Dyah Indrasari, Alida Roswita Harahap, Siskawati Suparmin, Windy Sahar, Renan Sukmawan

BACKGROUND: Light transmission aggregometry (LTA) and VerifyNow is commonly used to measure platelet responsiveness to clopidogrel. This study aimed to compare the results of LTA and VerifyNow P2Y12 assay for assessing the clopidogrel resistance in patients undergoing percutaneous coronary intervention and determine factors affecting clopidogrel resistance. METHODS: The subjects were 119 patients who underwent percutaneous coronary intervention (PCI) and had given loading dose of 600 mg clopidogrel. Blood samples were taken at 6 hour after clopidogrel loading dose. Platelet aggregation was measured by LTA and VerifyNow. RESULTS: LTA and VerifyNow assay showed fair agreement with Kappa=0.270, p=0.001. The proportion of resistance to clopidogrel using VerifyNow was 21.8% and LTA was 47.1%. Patients with diabetes melitus were more likely to develop clopidogrel resistance than patients without diabetes (OR of 7.67; 95% CI: 1.87-31.50; p=0.005). CONCLUSION: The ability of LTA and VerifyNow in detecting clopidogrel resistance were not comparable. Multivariate analysis results for VerifyNow shows diabetes mellitus as the greatest predictors of clopidogrel resistance.

178-85

Authors: Mahmood Thamer Altemimi, Alaa Khattar Musa, Abbas Ali Mansour

BACKGROUND: Obese women with polycystic ovary syndrome (PCOS) are at high risk for developing type 2 diabetes mellitus (T2DM). A baseline oral glucose tolerance test (2-h OGTT) is important to screen for dysglycemia in PCOS particularly those high risk women. Due to its advantages by fasting is not required and less day-to-day variability, glycated hemoglobin (HbA1c) might be a convenient screening tool. This study aimed to evaluate the performance of HbA1c vs. 2-h OGTT in the diagnosis of glycemic disorders in PCOS and to evaluate the correlation between glycemic disorders, insulin resistance (IR), and anthropometric measures. METHODS: One hundred and twenty nine women diagnosed with PCOS according to the Rotterdam 2003 criteria in Basrah were included in the study. All subjects were examined for weight, body mass index (BMI) and waist circumference (WC). Then they were tested for fasting glucose, 2-h OGTT, HbA1c, and fasting insulin to assess IR. RESULTS: The result of 2-h OGTT test showed that there were 21 subjects (16.1%) showed to have IGT. The result of HbA1c test showed that 25 subjects (19.4%) were diagonised with prediabetes. Meanwhile FPG test result showed that 34 subjects (26.4%) were having IFG. The HbA1c examination showed an underdetected the diagnosis of T2DM (0.8%) and an overdeteced diagnosis of prediabetes (19.4%) (p=0.021) and at HbA1c 5.55% (37.2 mmol/mol), the specificity was (74.3%) and sensitivity (56.5%) to discriminate normal from abnormal glucose status. Ninety nine women (76.7%) were either overweight or obese and most of them had IR (76.8%). CONCLUSION: Screening of glycemic disorders is crucial for PCOS by using 2-h OGTT regardless of risk factor and HbA1c seems to be an unsatisfactory screening tool to predict glycemic disorders in women with PCOS.

192-200

Authors: Anggi Laksmita Dewi, Dewi Kartikawati Paramita, Jajah Fachiroh

BACKGROUND: Single nucleotide variations (SNV) have been mapped to be associated with several human conditions and diseases. To validate the association between SNV to certain human traits or diseases, a large number of subjects must be included. Thus, in need of a fast, relatively economic, and reliable genotyping method. This can be achieved through the use of tetra-primer amplification refractory mutation system polymerase chain reaction (Tetra-primer ARMS PCR). This study reports strategy to develop Tetra-primer ARMS PCR-based genotyping of CHRNA3 rs8040868. METHODS: The optimization of Tetra-primer ARMS PCR was done through these steps: identification of gene sequence and position of single mutation; designing outer and inner PCR primers; amplification of target gene fragments through PCR by using outer primer; confirming genotype of the PCR product by using sequencing; determining an optimum ratio of outer and inner primer; and determining optimum annealing temperature and cycles for the PCR program. The PCR products were run in 2% gel agarose electrophoresis and visualized under UV illumination. RESULTS: Outer and inner primer ratio of 1:3 with annealing temperature of 64.4°C and 40x cycles was found to be the most optimum condition. Tetra-primer ARMS PCR was able to confirm the results of the DNA sequence of 2 samples, confirming wild-type variants (TT allele) and the heterozygous mutant (CT allele). CONCLUSION: Tetra-primer ARMS PCR was able to genotype rs8040868 of the CHRNA3 gene.

208-15

Authors: Dwi Wahyuni, Joko Waluyo, Jekti Prihatin, Fersty Isna Kusumawardani, Aditya Kurniawan

BACKGROUND: Traditional medicine that might be useful to treat typhoid fever is earthworms (Pheretima javanica K.). It is important to know whether any substances contained in Pheretima javanica K. extract are toxic and have pharmacological effects, which can be observed from the eosinophil and immunoglobulin E (IgE) levels in the blood. METHODS: Thirty Rattus norvegicus B. were included in this study and were separated into 5 groups, with 6 rats in each group. T1, T2, T3, and T4 were groups of rats treated with 50, 300, 2000, and 5000 mg/kg BW of Pheretima javanica K. ethanol extract granules (PJEEG), respectively, while the control group was a group of rats treated with 0.36 mg Ciprofloxacin. The eosinophil number and the IgE concentration were detected. Physical observations were also conducted to detect any allergic symptoms, toxicity symptoms, or histopathological changes. RESULTS: Administration of PJEEG significantly increased the number of eosinophil in every group, from 49.82 cells/μL to 259.26 cells/μL on average (p<0.001). PJEEG did not significantly affect IgE levels (p=0.624). The results showed that the administration did not cause any allergic and toxicity symptoms. Histopathological analysis of rats’ kidneys and liver also showed that the administration had no significant effect on histopathological changes. CONCLUSION: The administration of PJEEG does not cause allergic symptoms, toxicity symptoms, and histopathological changes in rats. The administrations also significantly increase eosinophil number in rats’ blood. However, the concentration of IgE in rats’ blood was not significantly changed after administration of PJEEG.

19-26

Authors: Miftakh Nur Rahman, Ajeng Diantini, Miswar Fattah, Melisa Intan Barliana

BACKGROUND: There is a continuous rise in the prevalence of central obesity and become a pressing health problem in the world. Central obesity followed by many metabolic disorders especially Type 2 Diabetes Mellitus (T2DM). The pathogenesis started from overnutrition signal that force pancreatic beta cells to produce a large number of insulin. Low-grade chronic inflammation that occurred also affects the organs sensitivity against insulin and caused beta cells to compensated this situation and at the end become exhausted and loss its function. CONTENT: Along compensation mechanism, certain nutrients were support the beta cells to maintain their mass and function to produce insulin. Short chain fatty acids (SCFAs) are gut microbiota fermentation product that act as nutrient and give an advantage to the proliferation and survivability of the beta cells. Zinc (Zn) also plays an important role in every step of insulin production. Moreover, these nutrients protecting pancreas against inflammation and oxidative stress through certain mechanism. Most of patients with central obesity are unaware of the presence of this disturbance at early stage. Whereas, at molecular level there is a magnitude of SCFAs and Zn level in the blood that would become an early signal and predict the damage of beta cells. SUMMARY: Quantification of these two nutrients in the blood expected to provide an early warning tool to maintain insulin adequacy and predict the possibility of beta cell failure in central obesity with promising performance.

34-9

Authors: Aileen Clarissa Dauhan, Aridamuriany Dwiputri Lubis, Munar Lubis

BACKGROUND: Early detection and treatment of sepsis can prevent septic shock and reduce mortality rate. Troponin can become a prognostic factor in sepsis. However, not all health facilities are equipped to assess troponin levels. Vasoactive-inotropic score (VIS) is a simpler and more accessible method to describe hemodynamic status. The aim of this study was to assess the suitability of VIS score as early prognosis and mortality predictor of sepsis METHODS: A retrospective study was conducted to determine the correlation between VIS and troponin levels for sepsis cases in Pediatric Intensive Care Unity (PICU) Haji Adam Malik Hospital, Medan from January 2018 to December 2019. VIS score at 48 hours, maximum VIS score, pediatric logistic organ dysfunction-2 (PELOD-2) score, cardiac troponin levels at 48 hours were taken from medical records. RESULTS: There were 54 samples analyzed. VIS scores were positively correlated (p<0.001) to troponin T and troponin I levels at 48 hours (r=0.670 and r=0.606, respectively). VIS at 48 hours and maximum VIS were related to mortality (p=0.001 and p<0.001, respectively). VIS score at 48 hours could be used as a predictive factor for mortality (area under the curve (AUC): 79.7%, p<0.001) with a cut-off point at 11 (74.4% sensitivity and 80% specificity). High VIS at 48 hours indicated poor outcomes of sepsis in children with odd ratio (OR) value: 1.99 (95% confidence interval (CI): 1.25-3.19). CONCLUSION: Vasoactive-inotropic score was suitable as an alternative to cardiac troponin T and troponin I levels at 48 hours to early detect myocardial dysfunctions and mortality in children.

48-54

Authors: Mardiah Suci Hardianti, Syahru Agung Setiawan, Nungki Anggorowati, Wiwiek Probowati

BACKGROUND: Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease with dysregulation of somatic hypermutation (SHM) and class switch recombination (CSR) have been known to contribute for its lymphomagenesis. Activation-induced cytidine deaminase (AID) enzyme plays a vital role for both processes. Multiple myeloma oncogene 1 (MUM1) is known to upregulate the AID expression in normal and pathological conditions. However, both AID and MUM1 expression association in DLBCL is still unexplored using immunohistochemistry method. We examined DLBCL samples and then retrospectively tested its correlation with clinical findings. METHODS: A retrospective cohort study with 20 cases of DLBCL biopsy tissue with AID and MUM1 antibody was conducted. The samples were then classified into concordant (AID+/MUM1+ or AID-/MUM1-) and discordant group (AID-/MUM1+). The clinicopathological comparison was performed to observe any association between immunohistochemistry expression and clinical findings. RESULTS: Among 20 samples of DLBCL, concordant expression rate of AID and MUM1 was 80% with kappa Cohen’s of 0.578 (p=0.004). A significant association was observed between AID and MUM1 expression with a prevalence ratio of 2.25 (95% CI: 1.08-4.67; p=0.008). Clinical characteristics were not significantly different between each group. Restricted mean survival time was shorter in the concordant group compared with the discordant group but statistically insignificant (21.16 vs. 22.5 months; p=0.531). CONCLUSION: The result of this study showed the association between AID and MUM1 expression in DLBCL. However, whether the association may add further molecular heterogeneity of DLBCL is still to be confirmed by expanding the study.

61-7

Authors: Novalia Chumaladewi Guntarno, Anny Setijo Rahaju, Nila Kurniasari

BACKGROUND: Bladder cancer is the tenth-most common cancer in worldwide and the majority are urothelial carcinomas. The depth of invasion plays important role in the prognostic and therapeutic factor in urothelial carcinomas. The expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) may associated with the progression of bladder carcinoma, such as depth of invasion. In this study, the correlation between these two markers in urothelial bladder carcinoma invasion will be analyzed. METHODS: An analytical observational research with cross-sectional were conducted on 54 formalin fixed paraffin-embedded tissue from radical cystectomy (RC) which were diagnosed as bladder urothelial carcinoma at the Anatomical Pathology Laboratory of Dr. Soetomo Academic Hospital, Surabaya and divided based on the T stage were immunostained using VEGF and MMP-9 monoclonal antibodies. The difference of VEGF and MMP-9 expression in T stage of bladder urothelial carcinoma were analyzed using Kruskal-Wallis and Anova test, the correlation between VEGF and MMP-9 expression in various T stage of bladder urothelial carcinoma were analyzed using Spearman test. RESULTS: This study showed no significant difference of VEGF expression among T stage of bladder urothelial carcinoma (p>0.05) but there was significant difference of MMP-9 expression in T stage of bladder urothelial carcinoma (p=0.043). There was a correlation between VEGF and MMP-9 in various T stage of bladder urothelial carcinoma (rs= 0.50, p=0.001). CONCLUSION: The significant correlation of VEGF and MMP-9 expression in bladder urothelial carcinoma may prove the synergistically role of both proteins in tumor invasion by MMP-9 degradation extracellular matrix.

75-83

Authors: Erwin Astha Triyono, Sarah Firdausa, Heru Prasetyo, Joni Susanto, James Hutagalung, Lilis Masyfufah, Budiono Budiono, Ivan Hoesada

BACKGROUND: Human Immunodeficiency Virus (HIV) is an infectious disease that targets the human immune system by attacking cluster of differentiation (CD)4 cells. The use of propolis in HIV patients is expected to be safe and beneficial in terms of increasing endurance and immunity by its role in increasing CD4 level. This study aimed to analyze the influence of propolis supplementation in increasing the CD4 level in anti-retroviral (ARV)-treated HIV patients. METHODS: Double-blind randomized controlled clinical trial was conducted in 50 HIV patients who took regular ARV therapy. The subjects were divided into two groups, one group was treated with ARV and propolis, while another one was given ARV and placebo. The CD4 cell count was measured during pre-treatment, in the 3rd month, in the 6th month after treatment. The level of hemoglobin, leukocyte, and platelets were also measured. The SF-12 questionnaire was used to evaluate quality of life of the subject. RESULTS: Out of 50 subjects, 43 subjects completed the study, which were 19 subjects from the propolis group and 24 subjects from the placebo group. After 3-month of treatment, there was a statistically significant difference in the increase of CD 44 level in propolis group, while the increment was not significant in the placebo group. After 6-month treatment, the increase of CD4 level was occurred in both groups, propolis and placebo, however the increment was not statistically significant. The levels of hemoglobin, leukocyte, and platelets were not altered by the treatment and remained normal throughout the study. The quality of life was improved during the study; however, it was also not statistically significant. Mild adverse events occurred in 3 subjects which were relieved after the treatment stopped. CONCLUSION: Based on the result of this study, the administration of propolis on HIV patients receiving ARV bring significant difference in the increase of CD4 in propolis group from baseline to 3 month after the treatment. While in placebo group, this increment was not significant. At the end of study, CD4 count continued to rise up, however the increase was not statistically significant. There are no hemoglobin, leukocyte, platelets, and quality of life abnormalities. Therefore, it is necesary to do further research with a spesific CD4 count. However, it may be beneficial in relieving the clinical symptoms and quality of life of patient living with HIV.

91-6

Authors: Diana Nur Afifah, Fauzia Purnamasari, Luthfiatul Khusna, Noviasti Rahma Utami, Aida Fitri Nazillah, Syafira Noor Pratiwi, Fillah Fithra Dieny, Aryu Candra, Ayu Rahadiyanti, Rachma Purwanti, Enny Probosari, Martha Ardiaria, Nyoman Suci Widiastiti, Ferry Sandra

BACKGROUND: Administration of resistant starch (RS) influences the diversity and the composition of microbiota as well as inhibits the growth of cancer cell. Banana as a potential source of RS has been reported. Although Musa paradisiaca has been reported to induce apoptosis in colon cancer cells, Musa balbisiana, which has low glycemic index and suitable for particular patients, has not been investigated yet. METHODS: Starches of M. balbisiana and M. paradisiaca were prepared and mixed with other components to make 3 types of mouse pellets. Mouse model for colon cancer was prepared and fed with different types of mouse pellets. Blood was collected and processed for measuring β-glucuronidase and malondialdehyde (MDA) with Enzyme-linked Immunosorbent Assay (ELISA) method. Resected ceca were incised to collect the inner part for short-chain fatty acid (SCFA) measurement with gas chromatography analysis. Resected colas were fixed and processed for immunohistochemistry to detect Caspase-3. RESULTS: Colon-cancer-mice fed with the M. balbisiana and M. paradisiaca starches-contained pellets had significant higher concentrations of total SCFA (p=0.003), acetic acid (p=0.000), propionic acid (p=0.000) and butyric acid (p=0.000); lower concentration of β-glucuronidase (p<0.001); higher Caspase-3 score (p=0.040); and lower MDA concentration (p<0.001) than colon-cancer-mice fed with standard pellet (control). CONCLUSION: M. balbisiana and M. paradisiaca starches could be suggested as potential anti-colon cancer RS. Further research should be carried out to disclose the starches mechanisms in colon cancer cell.

304-12

Authors: Marwa Salah Mahmoud

BACKGROUND: Valproate sodium is an anticonvulsant drug. Saussurea lappa (costus) is a medicinal plant rich with antioxidants. This research aimed to assess the protective effect of costus root extract against valproate sodium-induced thyrotoxicity. METHODS: Eighty adult male albino rats were equally divided into four groups; group I: untreated control, group II: rats were given 200 mg/kg BW valproate sodium orally and daily for 8 weeks, group III: rats were given 300 mg/kg BW costus root extract orally and daily for 8 weeks, and group IV: rats were given combination of valproate sodium and costus root extract. After 8 weeks, blood samples were collected to evaluate T3, T4 and thyroid-stimulating hormone (TSH) levels. Thyroid gland samples were handled for light and electron microscopic investigation. The heights of follicular cells, area % of collagen fibers and color intensity of thyroglobulin immunoreaction were statistically analyzed. RESULTS: After being given valproate sodium as an induction, hormonal assay showed significant decrease in serum T3 and T4 and significant increase of TSH. Follicular and cellular alterations were shown by light and electron microscopes. Morphometric study revealed increased follicular cell height and area % of collagen fibers and decreased color intensity of thyroglobulin. In contrast, costus root extract appeared to have a protective role against valproate sodium-induced thyroid injury. Most of the changes induced by valproate sodium were not observed after supplementation with the plant root extract. CONCLUSION: Valproate sodium has serious effects on the function and structure of thyroid gland, and this study shows that costus root extract could have a protective effect against these effects.

320-4

Authors: Muhammad Hanif Wibowo, Riri Andri Muzasti, Syafrizal Nasution

BACKGROUND: Vascular calcification contributes greatly to the incidence of cardiovascular disease. Previously, vascular calcification was considered as a passive process caused by the mineral deposition from the circulation. Nowadays, researchers have found inhibitors and promoter factors from vascular calcification, one of which is the matrix gla protein (MGP). MGP levels depend on the gene that encodes them. The MGP T-138C polymorphism is one of the most common causes of vascular calcification. METHODS: This was a case-control study involving 86 chronic kidney disease (CKD) patients who underwent regular hemodialysis in Rasyida Kidney Hospital, Medan, Indonesia. Vascular calcification was determined from lateral and posteroanterior abdominal X-ray. The MGP T-138 C polymorphism was analyzed using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). RESULTS: The TT/TC genotype associates with the incidence of vascular calcification with OR of 3.52, 95% CI: 1.23-10.106 (p-value<0.001). CONCLUSION: There is an association between MGP T-138C polymorphism, particularly on T Allele with the incidence of vascular calcification in CKD patients undergoing regular hemodialysis.

333-9

Authors: Septelia Inawati Wanandi, Dwi Retna Lestari, Noza Hilbertina, Nurjati Chairani Siregar, Sri Widia Jusman, Murdani Abdullah

BACKGROUND: Cancer-associated fibroblast (CAF) is the most abundant tumor stroma. Our previous study has demonstrated that the secretomes of CAF isolated from colorectal carcinoma (CRC) patients could induce epithelial-mesenchymal transition in the HT-29 CRC cell line. However, the role of CAF secretomes in CRC stemness is needed to be further investigated. Therefore, the present study aimed to investigate the effect of CAF secretomes from CRC patients on the expression of stemness markers in HT-29 CRC cells in comparison with the secretomes from normal fibroblasts. METHODS: Fibroblasts were isolated from tumor (CAF) and their counterpart non-tumor (NF) areas of three CRC patients undergone surgical resection. Normal preputium fibroblasts (PF) were isolated during circumcision of three healthy boys aged 8 years. All fibroblasts were grown in free-serum culture medium for 24 hours to collect 50% (v/v) conditioned medium (CM). Then, CM was supplemented to HT-29 CRC cells for 72 hours. The effects of CAF- and NF-CM on the mRNA expression of CD44, CD133, OCT4, and ALDH1A1 were analysed using qRT-PCR. Cells proliferation was measured using the trypan blue exclusion assay. RESULTS: Supplementation of CAF-CM (50% v/v) significantly increased CD44, CD133, OCT4, and ALDH1A1 mRNA expressions compared to that of NF-CM and control without supplementation but had no effect on the proliferation of HT-29 cells. CONCLUSION: CAF secretomes from CRC patients upregulate the expression of CRC stemness.

349-54

Authors: Ibnu Purwanto, Didik Setyo Heiyanto, Ahmad Ghozali, Irianiwati Widodo, Iwan Dwiprahasto, Teguh Aryandono, Sofia Mubarika Haryana

BACKGROUND: Triple-negative breast cancer (TNBC) represents a heterogenous disease which differ in characteristic, treatment response and prognosis. We aim to perform in-depth analysis on the clinicopathologic feature and the prognostic value of basal-like and non-basal-like TNBC patients in an Indonesian tertiary hospital. METHODS: We retrospectively included patients diagnosed with TNBC between 2014-2017. Clinical variables were collected from medical record. Expression of epidermal growth factor receptor (EGFR), cytokeratin 5/6 (CK5/6), p53 mutant and androgen receptor (AR) were examined by using immunohistochemistry (IHC). RESULTS: We included 67 subjects, 67.1% were basal-like and the remaining 32.9% were non-basal-like, with mean age of 51 years old, 59.7% subjects had BMI <25 and 40.3% subjects had BMI ≥25; 16.4%, 65.7%, and 17.9% subjects presented with early stage, locally advanced stage, and distant metastasis respectively; T<5 cm was found in 29.9% subjects, while 70.1% subjects had T≥5; 67.2% subjects presented with N-, while 32.8% subjects were N+. The most common histological type was infiltrating ductal (82% of subjects). P53 mutant and AR expressions were positive in 44.8% and 15% subjects, respectively. Basal-like subtype presented with younger age at and had higher expression of AR, while non-basal-like subtype is associated with BMI ≥25 (p<0.05). Basal-like subjects had shorter overall survival (23.9 months (95% CI: 21.9-25.9) vs. 26.1 months (95% CI: 23-29.2). CONCLUSION: Basal-like subtype is associated with worse prognosis, younger age at diagnosis and increased expression of AR, while non-basal-like subtype is associated with higher BMI in Indonesian TNBC.

361-7

Authors: Raden Tina Dewi Judistiani, Sefty Mariany Samosir, Setyorini Irianti, Benny Hasan Purwara, Budi Setiabudiawan, Johannes Cornelius Mose, Budi Handono

BACKGROUND: Cholecalciferol, hepcidin, and soluble transferrin receptor (sTfR) interaction play an essential role in iron hemostasis. Anemia in pregnancy contributes to morbidity and mortality both for the mother and baby. In this study, we assessed the correlation between hepcidin, sTfR and cholecalciferol in third trimester maternal anemia. We aimed to find the cut-off for hepcidin and sTfR. METHODS: A case-control study involving 56 pregnant women in each anemia and healthy group was nested on a previous larger cohort study in Indonesia. Serum hepcidin, sTfR and cholecalciferol level were measured by enzyme-linked immunosorbent assay (ELISA) method. RESULTS: Serum hepcidin and sTfR level were significantly higher in case group, while serum cholecalciferol level has no difference between the two groups. New cut-off points were found for hepcidin (<15.93 ng/mL) and sTfR level (>2234.45 ng/mL). Low level of hepcidin (OR=5.32) and high level of sTfR (OR=8.28) increase the risk of anemia. High level of sTfR (adjusted OR=4.725; CI 95%=1.730-12.904; p=0.02) was the most important factor contributes to anemia, followed by the low level of hepcidin (adjusted OR=3.677; CI 95%=1.363-9917; p=0.01). CONCLUSION: The high level of sTfR is the most important factor related to anemia in the third trimester, followed by the low level of hepcidin. Low cholecalciferol level tends to favor the incident of anemia. The new cut-off point of third trimester sTfR and third trimester hepcidin were established in this study and may be useful for risk assessment and treatment monitoring for anemia in pregnancy.

376-83

Authors: Diah Tri Utami, Nadzifa Nugraheni, Riris Istighfari Jenie, Edy Meiyanto

BACKGROUND: The presence of adverse side effects limits the use of doxorubicin (Dox) despite its cost-effectiveness compared to other chemotherapeutic agents. Brazilein (Be), the major compound of Caesalpinia sappan, performs co-chemotherapeutic potency in several cancer cell lines. This study evaluates the chemosensitizing effects of Be to Dox on colon cancer cell line, WiDr. METHODS: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was conducted to evaluate the cytotoxic effect of Be and its combination with Dox. The synergistic effect of Be and Dox was examined by using the Combination index (CI) parameter. Cell cycle and apoptosis profiles were done using flow cytometry with propidium iodide (PI)/RNase and Annexin V staining, respectively. RESULTS: The combination of Dox and Be at half of IC50 on WiDr cells showed a synergistic effect with a combination index of 0.4. Analysis of the cell cycle revealed that the combination caused cell cycle termination at the S and G2/M phase. This finding corresponded with the data that single treatment of Dox and Be induced cell cycle arrest at the different phases, namely S and G2/M phase, respectively. However, the combination treatment for 24 hours did not induce apoptosis. This combination should be further clarified as there was a possibility that many cells may underwent permanently arrest that halts to proceed apoptosis. CONCLUSION: Our findings suggested that Be synergizes with Dox to suppress the growth of WiDr cells via cell cycle arrest, hence, Be is potential to be developed as a co-chemotherapeutic agent. Our findings suggested that Be synergizes with Dox to suppress the growth of WiDr cells via cell cycle arrest, hence, Be is potential to be developed as a co-chemotherapeutic agent.

201-13

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Hypothalamus is the master regulator of body’s systemic homeostasis including energy balance, body temperature, sleep, blood pressure, and circadian rhythms. This review article will highlight the shifting of the old paradigm of obesityinflammation-metabolic syndrome, which was focused on visceral organs and systemic inflammation, into a new model involving microinflammation in the master regulator of endocrine system, i.e., hypothalamus. CONTENT: Since the early stage of over-nutritional conditions and aging process, microinflammation in hypothalamus has started to emerge, due to the activation of several proinflammatory signaling pathways, especially the nuclear factor kappa B (NF-kB) and c-Jun N-terminal kinase (JNK)-mediated nuclear transcriptional programs. Together with intracellular organelle stress signals, these pathways develop a chronic microinflammatory environment in the hypothalamus leading to obesity and metabolic disorders. SUMMARY: Hypothalamic inflammation has been noted not only as an important driver of impaired energy balance, but also contribute in altered neurocircuit functions and promote obesity-associated metabolic impairment.

220-6

Authors: Ronny Lesmana, Setiawan Setiawan, Ferdyan Efza, Yuni Susanti Pratiwi, Gilang Muhamad Nur Iqbal, Hanna Goenawan, Nova Sylviana, Unang Supratman

BACKGROUND: Ethionamide usage as one of the drug regimens still becomes a challenge due to high numbers of patients developing hypothyroid. Ethionamide had been associated with the inhibition of thyroid hormone (TH) synthesis and interestingly, ethionamide (C8H10N2S)-induced hypothyroidism is supported by its similar structure with thioamides, propythiouracil (C7H8N2S). However, hypothyroidism is not solely caused by its production, it could be caused by signaling alteration. Therefore, knowing that important TH action is determined via genomic pathway, alteration of this receptor could bring serious clinical problem. Unfortunately, there is limited study about the regulation of ethionamide and its connection on TH genomic signaling especially thyroid hormone receptor (TR) gene expression in soleus, gastrocnemius and cardiac muscle. METHODS: Thirty-eight rats were divided into control, ethionamide and propylthiouracyl groups. After 12-week treatment, rat were sacrificed, then gastrocnemius, soleus and cardiac muscles were dissected out, snap freezed using liquid nitrogen, and stored in -80oC until use. RNA was extracted and run for reverse transcription polymerase chain reaction (RT-PCR). RESULTS: In soleus muscle, ethionamide stimulated TR mRNA expressions and deiodinase compared to control group. In contrast, TRα1 gene expression was not affected by ethionamide administration. In gastrocnemius muscle, only TRβ1 gene and Dio2 gene expressions that were significantly increased compared to control group. In cardiac muscle, ethionamide significantly stimulated all the thyroid hormone receptor isoform and iodothyronine deiodinase gene expression compared to the control group. CONCLUSION: Long ethionamide treatment upregulates TR gene expressions and deiodinase in soleus and cardiac muscle, there is different expression pattern of soleus, gastrocnemius and cardiac muscle after ethionamide stimulation.

233-8

Authors: Johan Gunadi, Starry Homenta Rampengan, Janry Antonius Pangemanan, Agnes Lucia Panda, Nancy Lampus, Hasjim Hasbullah

BACKGROUND: Heart failure (HF) is a clinical syndrome caused by structural or functional cardiac disorders and is the final stage of every heart disease, marked by decreased functional capacity and patients’ quality of life (QoL). Suppression of tumorigenicity-2 (ST2) is a biomarker depicting heart fibrosis and remodeling that altered left ventricular geometry, which in turn decreases left ventricular contractility, decreases functional capacity, and ultimately affects the QoL of the HF patient. METHODS: An observational study was conducted with a cross-sectional approach involving 60 patients with systolic heart failure. Left ventricular geometry, left ventricular ejection fraction (LVEF), ST2 level, and other biomarkers were examined, continued by QoL assessment. RESULTS: The ST2 level (33.25±23.55 ng/mL) was negatively correlated with LVEF (r=-0.257; p=0.024) and was positively correlated with QoL (r=0.255; p=0.05). The LVEF was negatively correlated with QoL (r=-0.224; p=0.031). However, no significant correlation was found between left ventricular geometry with ST2 level or patients’ QoL. CONCLUSION: Elevated ST2 levels are correlated with decreased LVEF and worse QoL in systolic heart failure subjects. Therefore, ST2 together with LVEF can be used as prognostic tools for patients with HF.

245-50

Authors: Reza Gunadi Ranuh, Andy Darma, Alpha Fardah Athiyyah, Ergia Latifolia, Retno Asih Setyoningrum, Boerhan Hidajat, Siti Nurul Hidayati, Anang Endaryanto, Ferry Sandra, Subijanto Marto Sudarmo

BACKGROUND: Risk of acute respiratory infections in children less than 5 years of age is up to 95%. Zinc deficiency is one of the main risk factors. This study aimed to explore the effect of zinc on the bronchial mucosae inflammatory status expressed by nuclear factor (NF)-κB p105/p50, NF-κB p65, interleukin (IL)-8, and IL-1β. METHODS: Twenty-four Wistar rats were divided into 4 groups: normal zinc diet group without zinc supplementation (Z1), normal zinc diet group with zinc supplementation (Z2), zinc deficient diet group without zinc supplementation (Z3), and zinc deficient diet group with zinc supplementation (Z4). NF-κB p105/p50, p-NF-κB p65, IL-8, and IL-1β were measured by immunohistochemical staining. RESULTS: The inflammatory status of bronchial mucosae between Z1 and Z2 groups showed no difference (p=0.055). However, the inflammatory status of bronchial mucosae between Z3 and Z4 groups showed significant difference (p<0.01). Multivariate factorial design showed that zinc supplementation was beneficial when given to zinc deficient diet group with regard to decrease p-NF-κB p65, IL-8 and IL-1β levels (p<0.001) and increase dendritic cell (p=0.022). CONCLUSION: Zinc administration under conditions of zinc deficiency affects the inflammatory status, as shown by the decrease of p-NF-κB p65, IL-8 and IL-1β and the increase of NF-κB p105/p50.

261-6

Authors: Mas Rizky Anggun Adipurna Syamsunarno, Alif Bagus Rakhimullah, Uni Gamayani, Masahiko Kurabayasi, Tatsuya Iso, Ratu Safitri, Ramdan Panigoro

BACKGROUND: Iron-overload cardiomyopathy (IOC) is a major comorbidity in patients with chronic repetitive blood transfusion due to myocardial iron uptake that facilitated by calcium channels. As cardiac compensatory mechanism to IOC, we hypothesized the cardiac calcium channels expression would be increased and involved in cardiomyopathy progressivity. This study was aimed to investigate the gene expression of calcium channels in the heart of the iron overload mice model. METHODS: Mice were divided into three groups according to iron administration doses 0, 0.1, and 0.3 mg/day. Systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) were measured for the representation of cardiovascular outcomes. The heart tissues were harvested. Further mRNA levels of L-type calcium channels (LTCCs) and T-type calcium channels (TTCCs) were examined using semi-quantitative PCR. The expressions of cardiac calcium channels and blood pressure among the three groups were compared. RESULTS: The expressions of TTCCs in the two iron-injected groups were higher than the control group (p=0.018). The expressions of LTCCs were not different (p=0.413) among groups. SBP, DBP, and MAP of the iron-injected group were lower than the control group (p=0.025, p=0.011, and p=0.008, respectively). CONCLUSION: Iron administration affects the expression of TTCCs but not the LTCCs, accompanied by decreasing of systolic and diastolic blood pressure.

275-82

Authors: Deddy Satriya Putra, Fadil Oenzil, Eryati Darwin, Hafni Bachtiar, Tofrizal Tofrizal

BACKGROUND: Glutamine, a non-essential amino acid, is the main fuel in the gastrointestinal mucosa. It is thought to protect the intestinal mucosa against local or systemic injury from diarrhea. This study aimed to determine the relationship between glutamine supplementation and ileum histopathology in acute and chronic diarrhea rats induced by enteropathogenic Escherichia coli (EPEC). METHODS: A randomized post-test only control group design was conducted. Thirty Rattus norvegicus Wistar strain were divided into 5 groups: one negative control group, two acute, and two chronic diarrhea groups. All four diarrhea groups were induced by EPEC at a dose of 108 CFU/mL. One acute and one chronic groups were supplemented with glutamine at a dose of 810 mg/200 g body weight for 14 days. While the other two diarrhea groups were not treated. The intestinal histopathology of each group was assessed and the level of inflammation was classified. RESULTS: Significant differences in inflammation levels were found among the groups (p<0.05). The highest inflammation level was observed in the acute diarrhea group without glutamine supplementation. Inflammation levels of both acute and chronic diarrhea with glutamine supplementation groups were significantly lower than the inflammation levels of acute and chronic diarrhea without glutamine supplementation groups. CONCLUSION: Supplementation of glutamine reduces the level of inflammation and leads to the histopathological improvement of the rat’s ileum.

102-8

Authors: Munar Lubis, Aridamuriany Dwiputri Lubis, Badai Buana Nasution

BACKGROUND: Sepsis in children is a significant cause of morbidity and mortality in the pediatric intensive care unit (PICU). Assessment of pediatric sepsis using serial Pediatric Logistic Organ Dysfunction (PELOD)-2 score can be used as a prognostic factor. The use of biomarkers of sepsis is also used for diagnosis and predicting outcomes. Many studies have suggested that C-reactive protein (CRP) and procalcitonin (PCT) can be used to predict mortality. METHODS: A prospective cohort study was conducted to evaluate CRP, PCT, PELOD-2 score and its combination as a predictive factor of mortality in sepsis. All patients admitted to PICU Haji Adam Malik General Hospital, Medan, from April to November 2019 with suspected sepsis were included in this study. Blood examination and PELOD-2 scores were examined in the first 24 hours. RESULTS: A total of 79 children were included with a mortality rate 55.7%. The CRP, PCT, and PELOD-2 score were higher in nonsurvivor (2.8 (0.5-22.4) mg/dL; 9.36 (0.13-79.8) ng/mL; 9 (3-21), respectively). In multivariate logistic regression analysis, neither CRP nor PCT values could be independent predictors of mortality. The PELOD-2 score can be an independent predictor for mortality at a cut-off score of 7 (OR: 3.47 (95% CI: 1.68-7.19)). The combination of PELOD-2 and CRP scores as predictors of mortality showed lower values than PELOD-2 and PCT scores (0.80 vs. 0.95). The combination of all parameters only adds 1% of the predicted mortality value. CONCLUSION: PELOD-2 score with PCT value are recommended to predict mortality children with sepsis.

115-23

Authors: Ami Ashariati Prayogo, Andi Yasmin Wijaya, Winona May Hendrata, Steven Sheng Looi, Reny I’tishom, Lukman Hakim, Fedik Abdul Rantam, I Ketut Sudiana, Abdurachman Abdurachman

BACKGROUND: Cancer stem cells (CSCs) eradication might serve as a robust approach for cancer eradication. MCF-7 as breast cancer continuous cell line is known to contain breast CSCs (BCSCs) for its capability to maintain its original tumor population. CSCs enriched culture is a fundamental tool for CSCs targeted therapy development. Effective and unsophisticated CSCs dedifferentiation protocol for producing CSCs enriched culture is needed. METHODS: MCF-7 cells were cultured initially in Dulbecco's Modified Eagle Medium (DMEM) low glucose medium then changed to DMEM:F12. Serum starvation was performed during each medium refreshment gradually with fetal bovine serum (FBS) concentration of 10%, 5%, 2.5% until reaching 1% FBS concentration. Stable MCF-7 culture was then adapted to serum free culture system, containing DMEM:F12, epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and B27 supplement as dedifferentiation protocol for 18 days. Cluster of differentiation (CD)44 and CD24 double staining immunocytochemistry was performed to evaluate cell stemness. RESULTS: The population of cells expressing BCSCs markers (CD44+/CD24low) in non-adherent single cells subpopulation was significantly increased after the dedifferentiation procedure (70.39%) compared to control groups (0.71%) (p<0.05). In contrast, the expression of BCSCs marker in adherent single cells subpopulation and for both adherent and non-adherent mammosphere the BCSCs markers showed a stable expression. CONCLUSION: BCSCs enrichment of breast cancer cell cultures from MCF-7 breast cancer cell line can be performed. Breast cancer cell plasticity is observed during the dedifferentiation protocol. Development of dedifferentiation inducing protocols can serve as an important foundation for breast cancer therapy development through BCSCs elimination.

130-5

Authors: Ervin Monica, Primayuni Dhia Hasanah, Arief Fadillah, Rara Aulia, Eko Sulistijono, Satrio Wibowo

BACKGROUND: Anemia is frequently found in ulcerative colitis (UC) patients and assumed to be related to inflammatory process. Vitamin D3 (VD) is known to have anti-inflammatory and immunomodulatory effects. It also has the potential to be an alternative treatment of the inflammatory process that occurs at UC, however its mechanism has not been clearly established. This study aimed to assess the effect of VD on histopathology and hemoglobin levels in UC through its regulation in tumor necrosis factor (TNF)-α and interleukin (IL)-6. METHODS: Total samples of 24 mice were divided equally into Sham group, UC group, UC+VD group (given 3% dextran sodium sulfate (DSS) followed by VD), and VD+UC group (given VD followed by 3% DSS). Mouse Colitis Histology Index (MCHI) was used to measure histopathological changes. Immunohistochemical staining was used to observe expression of TNF-α and IL-6 in colon. Evaluation of anemia was determined by hemoglobin levels. RESULTS: Based on MCHI scores, significant epithelial damage was found in colon sample of UC group (8.25±3.05) compared to Sham (0.33±0.26), UC+VD (2.33±1.07), and VD+UC group (2.83±0.75) (p<0.05). Significant lower numbers of TNF-α were found in Sham (27.33±3.42), UC+VD (36.33±1.86), and VD+UC group (36.68±1.86) compared with UC group (44.66±4.87) (p<0.05). Significant less IL-6 expression was found in Sham (18.05±2.96), UC+VD (24.78±0.79), and VD+UC group (25.09±2.79) compared to UC group (38.85±3.51) (p<0.05). Differences in hemoglobin levels were significantly lower in UC group (11.85±0.97) compared to Sham (14.25±0.47), UC+VD (13.68±0.68), VD+UC group (13.52±1.07) (p<0.05). CONCLUSION: VD significantly reduced proinflammatory cytokines, increased mucosal repair, and improved hemoglobin levels.

143-8

Authors: Putu Nita Cahyawati, Desak Putu Oki Lestari, Ayu Savitri Siskayani, I Made Toya Ariawan

BACKGROUND: Statin is an anti-cholesterol drug that is widely prescribed throughout the world. Statins are mainly used to treat and prevent cardiovascular disease. Several studies have found the pleiotropic effect of statin. However, related effect of statin in kidney failure is still unclear. Ischemic-reperfusion (I/R) injury is a major cause of acute kidney failure. This study aims to determine the effect of simvastatin on kidney function and glomerular conditions by periodic acid-schiff staining in I/R injury. METHODS: Eighteen male Swiss mice were grouped into sham operation group (GSO), I/R injury group (GIRI), and simvastatin group (GSIM). The GSO group was performed by sham operation and pretreatment of 1% carboxymethylcellulose (CMC) for 3 days. The GIRI group was performed by I/R procedure and pretreatment of 1% CMC for 3 days and the GSIM group was performed by I/R procedure and pretreatment of 10 mg/kg BW simvastatin for 3 days. Blood urea nitrogen (BUN) and creatinine serum were assessed to determine kidney function. Histopathological analysis of glomerulosclerosis was assessed by the extent of glomerular damage (sclerosis), capillary loops, and synechia. The data were analyzed by one-way ANOVA followed by post hoc Tukey’s test (p<0.05). RESULTS: The creatinine and BUN levels in the GIRI group were the highest (0.97±0.48) compared with the other groups. The glomerulosclerosis index in the GSO group was 0.75±0.56, the GIRI group was 3.55±0.61, and the GSIM group was 2.08±1.37. There was a significant difference in the glomerulosclerosis index between the GSO and GIRI groups, but there was no significant difference between the GIRI and GSIM groups. These differences include the formation of sclerosis in the glomerulus, capillary loop, and synechiae. CONCLUSION: Simvastatin improves kidney function and glomerulosclerosis in I/R injury.

157-64

Authors: Merlyna Savitri, Ugroseno Yudho Bintoro, Made Putra Sedana, Muhammad Noor Muhammad, Pradana Zaky Romadhon, Putu Niken Ayu Amrita, Andi Yasmin Wijaya, Winona May Hendrata, Ami Ashariati Prayogo

BACKGROUND: Carbohydrate antigens 15-3 (CA 15-3) is a conventional tumor marker in breast cancer, with low sensitivity and specificity. MicroRNA (miRNA)-21 showed its stability in circulation and could serve as powerful biomarker. The aim of this study was to evaluate miRNA-21 as breast cancer biomarker compared to CA 15-3 in Indonesian population. METHODS: Circulating plasma miRNA-21 expression was measured using qRT-PCR in 49 patients at various stages of breast cancer and 16 healthy controls. The relative expression value of miRNA-21 was calculated using 2-ΔΔCt. Meanwhile, CA 15-3 was quantified using electrochemiluminescence immunoassay (ECLIA) methods. The results of miRNA-21 and CA 15-3 plasma circulating expression were compared with controls at each stage and between stages of breast cancer. RESULTS: CA 15-3 median level in breast cancer group was 1.60 times higher compared to control group (p=0.019), 21.00 m/mL and 13.05 m/mL, respectively. Median miRNA-21 expression in breast cancer group was elevated 4.92 folds compared to control group (p=0.001), 4.43 and 0.90, respectively. There was no significant difference of CA 15-3 level between controls and all stages of breast cancer group. CA 15-3 cut-off value was 15.05 m/mL (p=0.016) with 59.2% sensitivity and 62.5% specificity. Meanwhile, there was a significant difference of miRNA-21 expression between controls and most stages of breast cancer group. Circulating miRNA-21 expression cut-off value was 2.07 (p=0.000) with 91.8% sensitivity and 87.5% specificity. CONCLUSION: Circulating miRNA-21 expression and CA 15-3 levels were significantly increased in breast cancer group compared to control group. The miRNA-21 expression increased consistently with breast cancer stage progression. miRNA-21 could serve as superior biomarker compared to CA 15-3.

177-82

Authors: I Dewa Gede Ugrasena, Harianto Notopuro, Subijanto Marto Sudarmo, Ketut Sudiana, Djajadiman Gatot, Ponpon Idjradinata

BACKGROUND: Childhood acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy in Indonesia and often treated by methotrexate (MTX). Though it can be cured in 30-60% of patients, MTX resistance remains the major cause of treatment failure in childhood ALL. Previous sudies showed that its anti-leukemic property was moderated by MTX ability to inhibitmethylene tetra hydrofolate reductase (MTHFR) and thymidylate synthase (TS) in folate metabolism. This study investigates the correlation between MTHFR and TS polymorphism and MTX resistance in ALL children. METHODS: A total of 155 subjects obtained from all subjects prior to chemotherapy. DNA from blood samples were extracted and underwent polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) to evaluate MTHFR C677T and TS 5’-UTR 3R/3R polymorphism. RESULTS: There was significant correlation between MTHFR C677T and TS 5’-UTR 3R/3R gene polymorphism with MTX resistance. Subjectswith MTHFR C677T and TS 5’-UTR 3R/3R gene polymorphism were 4 times (p=0.007) and 6.4 times (p=0.001) more likely to be MTX resistant than those without gene polymorphisms, respectively. CONCLUSION: MTHFR C677T andTS 5’-UTR 3R/3R represent dominant gene polymorphism related to MTX resistance in childhood ALL.