The Indonesian Biomedical Journal

113-24

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Mesenchymal stem (stromal) cells (MSCs) have a multipotent character, able to differentiate into several cell types, thus MSC serve as a cell reservoir for regenerative medicine. MSC therapeutic potency more associated to their immunosuppressive and anti-inflammatory properties rather than the multipotency, by its mechanism to secrete soluble factors with paracrine actions. CONTENT: MSC paracrine function was known to mediated partly by extracellular vesicles (EVs), which were released predominantly from the endosomal compartment contained in MSC secretome. EV contain a cargo bring micro RNA (miRNA), messenger RNA (mRNA), and proteins from their cells of origin, propose EV as a novel alternative to whole cell therapies, regarding the benefit of EV in safety and easier storage compared to the parent cells. SUMMARY: The discovery of EVs including exosomes in MSC secretome as key of stem cells beneficial function lead to the future hope of using cell-free regenerative therapies.

136-44

Authors: Farzaneh Karimi, Sayyedehnikta Kasaei, Azar Baradaran, Farzaneh Ashrafi, Ardeshir Talebi, Zahra Lak, Mehdi Nematbakhsh

BACKGROUNDS: Cisplatin (CP) as an anticancer drug may affect the plasma glucose level while diabetic subjects are protected against CP-induced nephrotoxicity. In the current study, the role of dextrose hydration during CP therapy on CP-induced nephrotoxicity was evaluated. METHODS: Sixty-nine male and female rats were divided into 12 groups. The rats were hydrated with 15 mL/kg vehicle or different doses of 2%, 10% and 20% dextrose before and after 7.5 mg/kg CP administration. One week later, the biochemical and kidney function markers, and histology finding were determined. RESULTS: All the animals co-treated with CP and 20% dextrose, were dead during one week of the experiment. Administration of CP alone increased kidney tissue damage score (KTDS) and kidney weight (KW). It also elevated the blood urea nitrogen (BUN) and BUN-creatineine ratio (BUN/Cr) levels in the serum. In addition, CP decreased body weight and creatinine (Cr) clearance (ClCr) significantly in both male and female rats (p<0.05). However, 2% and 10% dextrose did not alter the mentioned parameters in male, but 10% dextrose supplement increased the serum levels of BUN, Cr and BUN/Cr ratio, KW and KTDS significantly in female rats (p<0.05). CONCLUSION: Our data suggest that not only do not support the nephro-protective role of dextrose hydration during CP therapy, the dextrose hydration can act as risk factor to promote CP-induced nephrotoxicity in female rats. Prohibition of high carbohydrate (glucose) diet during CP therapy is recommended.

152-8

Authors: Juminten Saimin, Hendy Hendarto, Soetjipto Soetjipto

BACKGROUND: Dyspareunia and pain due to the decrease of vaginal wall thickness usually happen in menopausal women. The reduction of estrogen levels cause the decreasing of cell proliferation and the thinning of vaginal wall epithelium. Tomato (Solanum lycopersicum) is a source of phytoestrogens, which produce estrogenic effects. This study aims to assess the effect of tomato juice on Ki-67 expression and epithelial thickness of the vaginal wall in menopausal rats. METHODS: This was an experimental study using Sprague-Dawley rats. Twenty-four female rats, aged 4 months and weighing 150-200 grams, were divided into 4 groups. Each group consisted of 6 rats. Negative control (NC) group was group of rats with sham procedure and performed by distilled water for 28 days. Positive control (PC) group was group of bilateral ovariectomized rats and performed by distilled water for 28 days. The first treatment (T1) group was group of bilateral ovariectomized rats, given tomato juice at dose of 11g/200g body weight (BW)/day. The second treatment (T2) group was group of bilateral ovariectomized rats, given tomato juice at dose of 15g/200g BW/day. Data analysis was done with Anova, multiple comparisons and regression test. RESULTS: The group with lowest Ki-67 expression was PC group (2.52±0.60). The expression of Ki-67 in treatment groups (T1 and T2) was higher than PC group, but lower than NC group. There was no significant difference between groups (p=0.771). The lowest epithelial thickness was found in PC group (21.19±3.96) and the highest was found in the treatment groups (38.73±12.43). There was positive correlation between tomato juice and epithelial thickness (p=0.647). CONCLUSION: Tomato juice increases Ki-67 expression and epithelial thickness on the vaginal wall of menopausal rats. The increase of epithelial thickness follows the administration dose, but Ki-67 expression does not exceed in the control rats. Tomato juice increases the cell proliferation of vaginal wall on menopausal rats, however the increase is still within normal limits.

167-74

Authors: Mohammed Ali Nashiry, Gabriele Ruth Anisah Froemming, Yeap Swee Keong, Aletza Binti Mohd Ismail, Aisha Mohd Din, Alyaa Mahmood Al-Khateeb

BACKGROUND: Hyperthermia is used as an adjuvant treatment to sensitize cancer cells to subsequent radiotheraphy or chemotherapy. The aim of this study was to study the effect of severe hyperthermia on osteosarcoma cells and its underlying causes. METHODS: Short-term (1 h) severe hyperthermia (45°C) was applied to osteoblast-like osteosarcoma cells (MG-63) and the heat shock response and cell death mechanisms were investigated after recovery at 37°C for 72 h. RESULTS: Cell viability was significantly reduced (p<0.05) and apoptosis was significantly induced by severe hyperthermia in MG-63 cells (p<0.001). Caspase 3/7, 4 and 12 activities were significantly increased after 72 h of recovery at 37°C, indicating that severe hyperthermia induced endoplasmic reticulum (ER) stress and apoptosis (p<0.05). Heat shock protein 90 alpha (Hsp90α) was significantly down regulated at the protein level after recovery, in association with apoptosis induction (p<0.01). Additionally, caspase 8 and 9 were activated, possibly as a result of ER stress or other stimuli while, B-cell leukemia 2 family protein (BCL-2) mRNA was down regulated (p<0.01). CONCLUSION: Severe hyperthermia could cause prolonged cell cytotoxicity by inducing apoptosis in association with inhibition of Hsp90α. This indicates the therapeutic potential of severe hyperthermia for the treatment of osteosarcoma.

182-7

Authors: Keri Lestari, Abdurahman Ridho, Nuning Nurcayani, Zelika Mega Ramadhania, Melisa Intan Barliana

BACKGROUND: The prevalence of diabetes mellitus is growing every year, including in Indonesia. Medicinal herbs were used empirically for lowering blood glucose. One of potential herb to have hypoglycemic activity is Stevia. Stevia rebaudiana Bertoni leaves contain stevoside, a natural, low-calorie sweetener that is 300 times sweeter than saccharose. In this study, we aimed to explore the hypoglycemic activity of S. rebaudiana Bertoni leaves extract in a rat model of type 2 diabetes mellitus (T2DM). METHODS: Male Wistar rats were feed high-fat, high-carbohydrate feed and sugar solution for 74 days to induce a diabetic rat model. The animals were then divided into five groups consisting of a negative control group treated with 2% Pulvis Gom Arabicum; a positive control group treated with Metformin 45 mg/kg body weight (BW); and three test groups treated with aqueous extract of S. rebaudiana Bertoni leaves at doses of 3.125, 6.25 and 12.5 mg/kg BW for 36 days. Blood glucose was measured on days 14, 28 and 36. RESULTS: The results showed that blood glucose levels over 36 days were significantly (p=0.043) lower in the group treated with S. rebaudiana Bertoni leaves extract. Further Newman-Keuls analysis suggested that the hypoglycemic activity of S. rebaudiana Bertoni leaves extract was dose-dependent. CONCLUSION: Our results indicate that S. rebaudiana Bertoni leaves extract has a potential role as a hypoglycemic agent in the treatment of T2DM.

194-9

Authors: Cityta Putri Kwarta, Heri Wibowo, Yordan Khaedir, Iris Rengganis, Hanny Siti Nuraeni

BACKGROUND: Allergic asthma is a degenerative atopic disease caused by allergic or hypersensitivity type-1. More than 50% of people with allergic asthma are caused by the presence of house dust mites (HDMs) allergens. METHODS: The cellular immunity response was evaluated through a peripheral blood mononuclear cell (PBMC) culture isolated from blood, using the ficoll gradient technique. Subjects were atopic asthma groups and non-atopic asthma groups. PBMC from each subject cultured was stimulated with HDMs allergen, then incubated in a CO2 5% incubator, 37o C for 72 hours. With the multiplex assay method, interferon (IFN)-γ, interleukin (IL)-13 and IL-10 were measured, meanwhile indoleamine 2,3-dioxygenase level (IDO) was measured by the enzyme-linked immunosorbent assay (ELISA) sandwich methods. RESULTS: The IFN-γ production in the supernatant of PBMC cultures was stimulated by phytohemagglutinin (PHA), Roswell Park Memorial Institute (RPMI) medium and allergens. The IFN-γ production in allergen-stimulated supernatants showed higher level of IFN-γ in the nonatopic group (4,681,455±3,434,851) than atopic group (4,363,300±2,067,941) even though it was not statistically significant (p=0.078). There were no differences between the mean of IL-13 production in atopic asthma group and non-atopic group. The IL-10 production in allergenstimulated supernatants was shown to be higher in nonatopic group and were statistically significantly different (p=0.015). The IDO production in allergen-stimulated supernatants was shown to be higher in the non-atopic group (272,231±269,564) than in the actopic group (13,273±400), and it was significantly different (p=0.007). CONCLUSION: Cellular immune profile of subjects with allergic asthma to Dermatophagoides pterronyssinus (Der p) is characterized by a type-2 inflammatory response that is dominant compared to type-1 inflammation (higher IL-13 ratio compared to IFN-γ) and to the role of anti-inflammation (higher IL-13 ratio compared to IL-10). The decline in IDO production in allergic asthma subjects to Der p is thought to be related to the low cellular immune response in expressing IFN-γ compared to IL-13.

205-9

Authors: Siska Darmayanti, Rini Hendriani, Cynthia Retna Sartika

BACKGROUND: Type 2 Diabetes Mellitus (T2DM) is a metabolic disease, due to the disorder of insulin function, insulin secretion, or both. Long-term hyperglycemia conditions promote endothelial dysfunction precedes to the development of multiple organ dysfunctions. Endothelial progenitor cells (EPCs) and hematopoietic stem cell (HSCs) are the key cellular effectors of postnatal neovascularization and play central role in endothelial dysfunction. However, in T2DM condition, the number of apoptotic HSCs increase, it may cause the reduction in potency and number of EPCs. In diabetes, the circulating EPCs number decrease and their functionality is impaired, but mechanism underlie of this impairement is unknown. The purpose of this study was to examine the relationship duration diabetes with the number and potency of EPC cells in T2DM patients controlled and poorly controlled. METHODS: Thirty-eight T2DM male patients were classified into two group based on Indonesian Society of Endocrinology/Perkumpulan Endokrinologi Indonesia (PERKENI) criteria on T2DM. The first group was a controlled glycemic condition group (hemoglobin A1c (HbA1C) <7.0%) and the second group was a poorly controlled glycemic condition group (HbA1C >7.0%). Cluster of differentiation (CD)34+ and CD133+ expressions were used as specific marker for EPC, while quantified bright aldehyde dehydrogenase (ALDHbr) assay was used to represented the potency of EPCs. RESULTS: This study showed that in poorly controlled T2DM group the number of EPCs was lower by 24.80% (p<0.05) compared to the T2DM controlled group. Similarly, the expression of ALDHbr was lower by 43.07% (p<0.05) in poorly controlled group. CONCLUSION: There was a decrease in the number and potency of EPCs in poorly controlled T2DM patients compared to the controlled T2DM patients. There was also a strong negative correlation between the duration of diabetes and number of EPCs.

217-24

Authors: Novi Silvia Hardiany, Yohana Yohana, Septelia Inawati Wanandi

BACKGROUND: Glioblastoma multiforme (GBM) is a human malignant brain tumor which is arise from glial cells. Our previous study proved that GBM cells proliferation increased after treating by conditioned medium of umbilical cord-derived mesenchymal stem cells (CM-UCSCs). Cells proliferation is probably mediated by tumor necrosis factor (TNF)-α which could bind to membrane receptor and induce signaling pathway. Therefore, this research was intended to analyze the mRNA expression of TNF-α signaling pathway molecules on CM-treated GBM cells by measuring TNF receptor 1 and 2 (TNFR1 and TNFR2), TNFR associated factor 2 (TRAF2), nuclear factor kappa B (NF-κB) mRNA level, and TNFR2 protein level. METHODS: UCSCs and human glioblastoma T98G cells were cultured and harvested after 80% confluence. CM was prepared by growing UCSCs in serum alpha Minimum Essential Media (α-MEM) for 24 hours. Fifty percent concentration of CM-UCSCs was used to treat T98G cells for 24 hours. TNF-α level in CM-UCSC was detected using enzyme linked-immunosorbent assay (ELISA), while the expression of TNFR1, TNFR2, TRAF2 and NF-κB were detected using quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR), and TNFR2 protein level was detected using sandwich ELISA. RESULTS: TNF-α level was detected in CM-UCSCs 4.4 pg/mL. Moreover, the expression of TNFR1, TNFR2, TRAF2 and NF-κB were significantly 1.4-fold, 4.9-fold, 5.6-fold, 1.8-fold respectively higher in T98G treated cells than control. TNFR2 protein level in T98G treated cells was 11.57 pg/mg protein higher than control. CONCLUSION: The expression of molecules involved in TNF-α signaling pathway were up regulated in T98G cells treated by CM-UCSCs.

1-15

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Mitochondrial dysfunction known to be associated with most of human inherited disorders and diseases, including neurodegenerative disorders, cardiomyopathies, metabolic syndrome, muscle weakness, cancer, also obesity. CONTENT: Mitochondria charges for multiple anabolic and catabolic circuitries, as the main provider for adenosine triphosphate (ATP). Mitochondria also responsible for cellwide stress responses and control non-apoptotic cell death routines, such as autophagy and regulated necrosis. In other words, mitochondria play an extended role in regulating cellular functions, both vital and lethal, from physiological metabolism to stress responses and death to maintain adult tissue homeostasis. Furthermore, mitochondria are crucial for both embryonic and postembryonic development. Therefore, any defect or alteration in mitochondria signaling pathways will lead to a large number of diseases in human, including premature aging, neurodegenerative disorders, muscle weakness, cardiovascular disorders, and cancer. SUMMARY: Mitochondria perform a dynamic, integrated interconnected network, to maintain tissue homeostasis, beyond the cell boundaries and regulating cells and tissues communication. Certainly any mitochondrial dysfunction could direct to neurodegenerative diseases and metabolic disorders.

30-5

Authors: Sri Adi Sumiwi, Ade Zuhrotun, Rini Hendriani, Mochamad Rizal, Jutti Levita, Sandra Megantara

BACKGROUND: Previous works indicated various pharmacology activities of bay plant (Syzygium polyanthum (Wight) Walp.), however only few studies investigated its toxicity. This work was aimed to study the subchronic toxicity of ethanol extract of this plant. METHODS: White Wistar rats were divided into 4 groups and were treated with 2% of Arabic gum (PGA) suspension, 1000 mg/kg of body weight (BW), 400 mg/kg of BW and 100 mg/kg of BW, respectively. The animals were observed on their body weight, hematology, clinical biochemistry parameters, organ index and histopathology. RESULTS: Flavonoids, tannins, polyphenols, saponins, quinones, monoterpenes and sesquiterpenes were detected in dried leaves and ethanol extract of bay plant. An increase of body weight in male and female groups treated with dose 100 and 400 mg/kg BW compared to controls, was observed. Moreover, there was an increase of white blood cell (WBC) in male and female groups treated with S. polyanthum extracts compared to controls, whereas a decrease of red blood cell (RBC) was observed in male groups treated with S. polyanthum extracts in dose-dependent manner compared to control. No significant changes of RBC were seen in female groups, haemoglobin values were not altered by extract treatment. Photomicrographs of liver, kidney, lungs, heart and spleen histopathology of male and female S. polyanthum extract-treated groups showed no significant alteration compared to controls. CONCLUSION: Our study revealed that S. polyanthum extracts does not show toxicity on the body weight, hematology, creatinine and serum glutamic pyruvic transaminase (SGPT), but fatty liver and necrosis are observed in female rats. This result can be beneficial for plant-based drug discovery, particularly this study provides information about the safety of S. polyanthum to be further developed as candidate of phytopharmaceutics.

42-7

Authors: Mona Hmoud AlSheikh, Shayma Ibrahim Almubayadh

BACKGROUND: Vitamin D is an essential nutrient requirement for adults and children for normal metabolism and maintenance of healthy bones. Its deficiency may disrupt homeostasis and contribute to systemic diseases, especially in obese patients. This study aimed to examine the effect of vitamin D on insulin, fasting blood glucose (FBG) and waist-hip ratio (WHR) in young and obese Saudi females with pre-existing hypovitaminosis D. METHODS: The results of this study were based on prospective parallel randomised controlled clinical trial conducted at King Fahd University Hospital in which 30 obese Saudi young females aging 18-23 years with preexisting vitamin D deficiency contributed. Participants were randomly assigned to either vitamin D supplementation group (experimental) or placebo group (control). Subjects of vitamin D group received 50,000 IU/week of cholecalciferol drops and placebo group received 10 mL of normal saline drops for 12 weeks. RESULTS: The results of multivariate repeated measures analysis showed a change in serum 25-Hydroxycholecalciferol levels after vitamin D supplementation (F=296.132, p<0.05). There was also an indication of difference between the vitamin D group and placebo group. The only significant relationship was found between vitamin D and FBG (p<0.05). Supplementation of vitamin D improved FBG but did not contribute to improving insulin level. There was no association between vitamin D supplementation and WHR. CONCLUSION: This study concludes that supplementation of vitamin D improves FBG, but it has no contribution in improving insulin level. No association exists between vitamin D supplementation and WHR. The insignificant correlation of vitamin D intake with WHR and insulin can be attributed to the small sample size, therefore, further research is suggested with large sample size.

52-8

Authors: Supriono Supriono, Asri Nugraheni, Handono Kalim, Mudjiwijono Handaru Eko

BACKGROUND: Transforming growth factor (TGF)-β1 has a pivotal role in liver fibrogenesis. Curcumin effectively prevent the progression of liver fibrosis through inhibition of TGF-β1/Sma and drosophila MAD (Smad) signaling pathway. However, the role of curcumin in the regression of liver fibrosis is still unknown. This study investigated the role of curcumin and TGF-β1 in liver fibrosis regression. METHODS: An experimental Wistar rat model included 6 treatment groups as well as positive and negative control groups. The treatment and positive control groups were injected with carbon tetrachlorid (CCl4) for 9 weeks to induce liver fibrosis. After cessation of injection, 3 of the treatment groups were given curcumin and 3 were given carboxymethylcellulose (CMC) for 2, 5 and 9 weeks, while the positive control was untreated. The negative control was injected with normal saline. TGF-β1 liver tissue levels were analyzed by ELISA, while the TGF-β1 expression in liver cells was analyzed by immunohistochemical assay. The metavir score was used to assess the degree of liver fibrosis. Values of p<0.05 were regarded as statistically significant. RESULTS: Nine weeks of CCl4 injection induced liver fibrosis (metavir F3); and significantly increased TGF-β1 levels and expression in tissues (p=0.00, p=0.021, respectively). Curcumin administration decreased levels and expression of TGF-β1 in the liver and accelerated regression of liver fibrosis. There was a significant correlation between duration of administration of curcumin with an expression of TGF-β1 in the liver tissue (r=0.87; p<0.00). CONCLUSION: Curcumin accelerates regression of liver fibrosis, likely through decreasing of TGF-β1 expression in the liver.

63-9

Authors: Amelia Lorensia, Zullies Ikawati, Tri Murti Andayani, Daniel Maranatha, Mariana Wahyudi

BACKGROUND: Aminophylline (theophylline) is one of the most frequent asthma therapies in Indonesia, although it remains as a narrow therapy. The effects of drugs are individualized and strongly influenced by genetic, one of which is CYP1A2 gene polymorphisms. This study aimed to determine the profile of CYP1A2 polymorphism and theophylline level in asthma exacerbation patients receiving intravenous aminophylline therapy. METHODS: This cross sectional study was conducted in the emergency room (ER), to adults asthma exacerbation patients without complication (n=27), visiting the ER. The gene polymorphism data were compared with theophylline levels in the blood using chi-square test. RESULTS: In the CYP1A2 gene polymorphism profile, the most common heterozygous alleles are T/G genotype of CYP1A2*1E and C/A genotype of CYP1A2*1F. Most homozygote alleles exist in CYP1A2*1D and CYP1A2*1F. There was significant difference between CYP1A2*1D (p<0.005), CYP1A2*1E (p<0.023) and CYP1A2*1F (p<0.000) polymorphisms and theophylline level. CONCLUSION: CYP1A2*1D, CYP1A2*1E and CYP1A2*1F gene polymorphisms had an effect on theophylline levels. However, no one experienced an overdose theophylline, and no correlation between theophylline levels with CYP1A2 gene polymorphism.

78-84

Authors: Arum Kartikadewi, Awal Prasetyo, Lisyani Budipradigdo, Heri Nugroho, Kusmiyati Tjahjono, Arthur Lelono

BACKGROUND: The depletion of glucose transporter 4 (GLUT-4) affects blood glucose level in the type 2 diabetes mellitus (T2DM) patient. Herbals supplementation such as Artemisia annua is known to posses an antidiabetic potency, but its effect on long term glucose control marker, glicated hemoglobin (HbA1C) and muscle GLUT-4 expression still has not been investigated. METHODS: Twenty Wistar rats were divided into four groups (five rats per group). The C1 group was consist of healthy control rats and C2 group was consist of diabetic control rats. Diabetic condition was induced by giving high lard fat diet for 28 days, followed with 30 mg/kgBW of Streptozotocin injection at the 29th day. Meanwhile, T1 and T2 group were consist of diabetic rats, which were supplemented with 50 mg/kgBW and 100 mg/kgBW of Artemisia annua leaf extract, respectively. GLUT-4 expression and HbA1C level were measured at the 14th day post-treatment. RESULTS: The GLUT-4 was expressed 1.8 and two times higher in T1 and T2 group, respectively, compared to C2 group. Meanwhile, HbA1C level in C2 group was two times higher than C1 group (11.95±2.52 compared to 5.61±2.69, p<0.01). Further, the administration of 100 mg/ kgBW Artemisia annua extract caused a slight reduction of HbA1C (11.95±2.52 compared to 8.18±3.53, p>0.05) CONCLUSION: Muscle cell’s GLUT-4 in T1 and T2 group was expressed increasingly and significantly different compared to the C2 group. HbA1C level in T2 was slightly reduced although no significant different compared to C2 group.

91-9

Authors: Mahalul Azam, Susanti Lestari, Sri Ratna Rahayu, Arulita Ika Fibriana, Budhi Setianto, Nyoman Suci Widyastiti, Suhartono Suhartono, Hardhono Susanto, Martha Irene Kartasurya, Udin Bahrudin, Thijs Eijsvogels

BACKGROUND: Inflammation plays an important role in the atherosclerotic process. High-sensitivity C-reactive-protein (hs-CRP) is commonly used as inflammatory biomarker. It is well known that regular physical activity lowers hs-CRP levels, while prolonged exercise induces hs-CRP elevations. However, the relationship of training and exercise characteristics with hs-CRP levels remains not well elucidated. We evaluated baseline and post-exercise hs-CRP levels and its association with training and exercise characteristics. METHODS: Eighty-eight male endurance cyclists were involved. Demographic data, health condition and training characteristics were collected. Baseline and postexercise blood-samples were collected to determine hsCRP concentrations. A hs-CRP cut-off point of 3 mg/L was used. Blood-cell count and biochemical parameters were measured at baseline. Heart rate (HR) was measured during exercise. RESULTS: Cyclists performed 7.3 hours (interquartilerange (IQR) = 5.4-7.5) of endurance exercise at intensity of 81.8 % (IQR = 74.9-85.8). Cyclists with baseline hsCRP ≥ 3 mg/L reported higher body mass, body mass index (BMI), waist-circumference and total-cholesterol. An increase in hs-CRP was following endurance exercise. Cyclists with any elevation of hs-CRP reported a higher BMI, HR during exercise and exercise intensity. Binary logistic regression analysis showed BMI (OR = 1.24, 95% CI = 1.04-1.48) and cycling distance (OR = 0.22, 95% CI = 0.06-0.76) were associated with post-exercise hs-CRP elevations. CONCLUSION: Body mass, BMI, waist-circumference, total- and HDL-cholesterol are associated with baseline hsCRP, whereas BMI and cycling distance were associated with hs-CRP elevations. These findings suggest that anthropometry parameters and lipid levels attributed to baseline hs-CRP, while anthropometry parameters and cycling intensity attributed to post-exercise hs-CRP elevations.

107-12

Authors: Lucky Hartati Moehario, Thomas Robertus, Anis Karuniawati, Rudyanto Sedono, Delly Chipta Lestari, Andi Yasmon

BACKGROUND: Antibiotic resistance has become a worldwide problem. Among Asia countries, Indonesia has high prevalence of multi-drug resistant organisms mainly due to Gram-negative bacilli Enterobacteriaceae. This study aimed to find out whether gene family of AmpC and AmpC/ESBL-producing Enterobacteriaceae were present in the Intensive Care Unit (ICU) of Cipto Mangunkusumo Hospital, Jakarta, Indonesia. METHODS: Specimens were obtained from several body sites of adult patients with infection hospitalised in ICU of Cipto Mangunkusumo Hospital. VITEK®2 was used to identify the microorganisms. Antibiotic susceptibility tests were conducted using VITEK®2 and disc diffusion technique according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Double disc synergy (DDS) test method was employed to detect AmpC activity. Gene families of ampC were identified using multiplex polymerase chain reaction (PCR). RESULTS: Forty five isolates were identified as putative AmpC, extended-Spectrum β-lactamases (ESBL) and AmpC/ESBL-producing Enterobacteriaceae. Klebsiella pneumoniae (n=32) were predominant, followed by Escherichia coli (n=6), Enterobacter cloacae (n=5) and Enterobacter aerogenes (n=2). AmpC activity was detected in 9 isolates, in which 4 isolates were AmpC producing and 5 isolates were AmpC/ESBL. In vitro, AmpC-producing Enterobacteriaceae showed good susceptibility to many antibiotic tested, while those of AmpC/ESBL-producing only to Amikacin. The gene families of ampC were DHA, EBC and CIT identified from 6 isolates. CONCLUSION: DHA, EBC and CIT gene families were identified from AmpC and AmpC/ESBL-producing Enterobacteriaceae in the ICU of Cipto Mangunkusumo Hospital. While the AmpC-producing was still susceptible to almost all antibiotics tested, the AmpC/ESBL-producing showed resistant except for Amikacin.

192-202

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Current understanding in molecular character of natural killer (NK) cell, its function and mechanisms, send people the ideas to develop a NK-cell-based immunotherapeutic strategies against human cancer. CONTENT: Before being regards as a cell-based cellular therapy, NK cell have to be clinical proven. Early studies with NK cells infusions for acute myeloid leukemia and lung cancer showed a promising result. NK cells need simplified methods for enriching and expanding, in addition to its transfection with chimeric antigen receptors (CARs). NK-92 arise as an assuring effector cells to augment monoclonal and bispecific antibody therapy. Thus, NK cells show a potential opportunity for cell engineering, outstep the era of T cells. SUMMARY: It is believed that NK cells bring a bright hope for future cancer immunotherapies, either alone or in combination as a harmonious therapy.

217-21

Authors: Emy Noerwidayati, Andaru Dahesihdewi, Osman Sianipar

BACKGROUND: Klepsiella pneumoniae, one of clinical isolates, is frequently found causative agent of hospital acquired infection. Currently, K. pneumoniae is found as extended-Spectrum β-lactamases (ESBL) producer, allowing it to become multidrugresistant. A clinical laboratory with limited facility needs a valid, reliable, inexpensive and simple laboratory test to control its infection and antimicrobial-resistancy. The aim of this study is to evaluate the diagnostic performance of a ESBL media to detect ESBL-producing K. pneumoniae. METHODS: An independent and blind comparative study of ChromID ESBL media and Double Disc Synergy Test (DDST) was conducted for detecting the clinical isolate of ESBL-producing K. pneumoniae. Clinical isolates of K. pneumoniae collected from the Clinical Laboratory of Dr. Sardjito Hospital were isolated. RESULTS: There were 103 clinical isolates of K. pneumoniae, which were isolated from urine, pus, blood, stool, cerebrospinal fluid, sputum, drain liquid, nasal sinus liquid, gastric wash, bronchi liquid, injury liquid and nasal swab. The number of true positive, true negative, false positive and false negative results were 74, 18, 9 and 2, respectively. Meanwhile, the sensitivity, specificity, positive predictive value, negative predictive value, likelihood ratio for positive result and likelihood ratio for negative result of the new ESBL media were 97.4%, 66.7%, 89.2%, 90%, 2.9 and 0.03, respectively. CONCLUSION: Since the new ESBL media and DDST results were similar, so the new ESBL media could be used for screening patients with clinical presentation that indicating a high suspicious of ESBL-producing bacteria.

231-5

Authors: Angliana Chouw, Bayu Winata Putera, Cynthia Retna Sartika, Ajeng Diantini

BACKGROUND: Umbilical Cord Blood (UCB) has been widely use as regenerative medicine due to the content of undifferentiated cell which have capability to do self-renewal and differentiation into various type of cell called stem cells. Recent studies show that UCB contains not only hematopoietic stem cell (HSC) but also others stem cell and progenitor cell such as endothelial progenitor cell (EPC) and very small embryonic-like stem cell (VSEL). It is beliefs that HSC and EPC shared the same progenitor. In this study, correlation between the cell number of HSC, EPC and VSEL is analyzed in umbilical cord blood as the source of stem cell for clinical application. METHODS: The cell number of HSC, EPC and VSEL is counted from cryopreserved UCB collected from 22 women delivered via cesarean section which already stored for more than 2 years in this study. Sample were incubated with antibodies such as cluster of differentiation (CD)34-phycoerythrin (PE)/CD45-fluorescein isothiocyanate (FITC), CD133/1 (anti-CD (AC)133)-antigen-presenting cell (APC) and, CD184 (C-X-C chemokine receptor (CXCR)4)-PE.Vio770 to detect the present of HSC, EPC and VSEL in UCB. Sample were analyze using flowcytometer BD FACS Canto II. RESULTS: The cell population of HSC and late-EPC is 0.009% and 0.01% of total cell in UCB. VSEL only represented 0.001% from total cell in UCB, showing the lowest number of cell population in UCB. The correlation between the cell number of HSC and EPC is r=0,483*, p=0.023) and between HSC and VSEL is r=0.510*, p=0.015. CONCLUSION: In this study, both EPC and VSEL have a significant positive correlation with HSC.

243-9

Authors: Betty Nurhayati, Ira Gustira Rahayu, Sonny Feisal Rinaldi, Wawan Sofwan Zaini, Ervi Afifah, Seila Arumwardana, Hanna Sari Widya Kusuma, Rizal Rizal, Wahyu Widowati

BACKGROUND: Oxidative stress is closely related to all aspects of cancer. Cosmos caudatus ethanolic extract (CCEE) has been proved to have antioxidant effect that inhibited cancer cell growth due to its bioactive compounds such as catechin, quercetin and chlorogenic acid. This study aimed to evaluate antioxidant and anticancer activity of CCEE and its compounds. METHODS: Total phenol was measured according to the Folin-Ciocalteu method. Catechin, quercetin and chlorogenic acid contained in CCEE were identified by high-performance liquid chromatography (HPLC). Antioxidant activity was evaluated by 2,2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS)-reducing activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, and ferric reducing antioxidant power (FRAP) activity test. The cytotoxic activity of CCEE was determined by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay on HeLa cells. RESULTS: The result showed that total phenol of CCEE was 181.64±0.93 µg Cathecin/mg extract. ABTSreducing activity test showed that catechin had the highest activity (2.90±0.04 µg/mL), while CCEE had moderate activity compared to other compounds. FRAP activity test demonstrated that catechin had the highest activity (315.83 µM Fe(II)/µg) compared to other compounds. DPPH scavenging activity of CCEE was 22.82±0.05 µg/mL. Cytotoxicity test on HeLa cell showed that CCEE had lower activity (inhibitory concentration (IC)50= 89.90±1.30 µg/mL) compared to quercetin (IC50 = 13.30±0.64 µg/ mL). CONCLUSION: CCEE has the lowest antioxidant activity compared to quercetin, catechin, and chlorogenic acid and has the lowest anticancer activity compared to quercetin. However, CCEE and its compounds has potential as antioxidant and anticancer properties.

256-62

Authors: Umi Solekhah Intansari, Harina Salim, Usi Sukorini, Adika Zhulhi Arjana, Muhammad Juffrie

BACKGROUND: Pathogenesis of severe dengue infection has not been elucidated. Immune complex of pre-existing antibodies and heterotypic dengue virus bind to FcγII (cluster of differentiation (CD32)) receptor (FcγIIR) on monocyte facilitates entry and replication of dengue virus. Aim of this study was to evaluate the expression of FcγIIR on monocytes in patients infected with dengue and in healthy subjects. METHODS: This study used a cross-sectional design that included patients infected with dengue who were hospitalized in Dr. Sardjito General Hospital, Panembahan Senopati Hospital, and Sleman Hospital, who met the inclusion criteria and selected consecutively. Examinations were completed using a lyse, no-wash method of flow cytometry. Computerized statistical analysis was conducted and was considered to be significant if p<0.05. RESULTS: Sixty-five study subjects were divided into healthy subjects (24 subjects) and patients with dengue infection (41 subjects). There were no significant differences in hemoglobin (Hb) and hematocrit (Hct) values between the groups, but differences were found in the number of leukocytes, absolute number of monocytes and platelet count (p<0.001, 0.002 and <0.001, respectively). The mean expression of FcγIIR monocytes in patients with dengue infection (208.77±32.06 median fluorescent intensity (MFI)) and the healthy subjects (124.03±47.76 MFI) with p<0.0001. CONCLUSION: The mean expression of FcγIIR monocytes in patients with dengue infection was higher than in healthy subjects.

270-7

Authors: Ika Nindya Kadariswantiningsih, Mochammad Thaha, Cahyo Wibisono Nugroho, Berliana Hamidah, Haerani Rasyid, Zaky El Hakim, Maulana Muhtadin Suryansyah, Mohammad Yusuf Alsagaff, Djoko Santoso, Maulana Antiyan Empitu, Yusuke Suzuki

BACKGROUND: Establishment of inexpensive clinical laboratory tests to evaluate inflammation and oxidative stress is urgently needed in the limited resources settings. This study aims to investigate the potential of complete blood count (CBC) parameters and non-fasting cholesterol profile parameters to describe inflammation and oxidative stress in chronic kidney disease (CKD) patients. METHODS: Measurement of CBC, non-fasting cholesterol profile, high sensitivity C-reactive protein (hs-CRP) and malondialdehyde (MDA) were performed in 71 CKD patients grouped into hemodialysis (HD) and non-hemodialysis (non-HD). Correlation analysis were performed to assess the potential of CBC and cholesterol profile to describe the level of hs-CRP and MDA. RESULTS: In the HD group, total cholesterol was moderately associated with hs-CRP while total cholesterol/HDL-C ratio, monocyte/HDL-C ratio, monocyte/LDL-C ratio, neutrophil/HDL-C ratio, neutrophil/LDL-C ratio, platelet/HDL-C ratio and platelet/LDL-C ratio were strongly associated with hs-CRP. In the non-HD group, only neutrophil/total cholesterol ratio and platelet/total cholesterol ratio that were associated with hs-CRP. Total cholesterol, monocyte/LDL-C ratio, neutrophil/LDL-C ratio and platelet/LDL-C ratio were moderately associated with MDA while total cholesterol/HDL-C ratio, monocyte/HDL-C ratio, neutrophil/HDL-C ratio and platelet/HDL-C ratio were strongly associated respectively with MDA in HD group. In the non-HD group, total cholesterol/HDL-C ratio, neutrophil/HDL-C ratio and platelet/HDL-C ratio were moderately associated with MDA in non-HD group while monocyte/HDL-C ratio was weakly associated with MDA. CONCLUSION: Some CBC parameters and non-fasting cholesterol profile such as cholesterol/HDL-C, monocyte/HDL-C, neutrophil/HDL-C and platelet/HDL-C ratio showed a potential to describe the inflammation and stress oxidative markers, given some strong associations with the level of hs-CRP and MDA. Further study is needed to assess whether this parameter represent long-term prognostic value among CKD patients.

284-9

Authors: Vera Amalia Lestari, Ika Agus Rini, Gita Widya Pradini, Edhyana Sahiratmadja, Herman Susanto

BACKGROUND: From all of human papillomaviruses (HPV) genotypes capable of causing cervical cancer, it is estimated that 70 percent are HPV-16 and HPV-18. HPV-16 can infect the tissues in single infection or together with other high-risk types of HPV, and the most common is with HPV-18. The origin of HPV can be identified by its phylogenetic tree. The aim of this study was to determine the phylogeny of HPV-16 and HPV-18 multiple infection in cervical cancer, whether both HPVs were from the same origin. METHODS: Cervical tissue biopsies (n=33) were obtained from Hasan Sadikin Hospital in the period of September to November 2016. HPV genotyping test was performed to confirm the HPV-16 and HPV-18 multiple infection. L1 gene of both HPV-16 and HPV-18 were sequenced for phylogenetic analysis. RESULTS: Phylogenetic analysis of L1 HPV-16 and HPV-18 showed the closest relationship with sequence from China and Thailand, respectively. CONCLUSION: HPV-16 and HPV-18 multiple infection of a cervical cancer patient from Dr. Hasan Sadikin General Hospital Bandung showed a very close L1 phylogeny relationship with isolate from Asian region.

297-302

Authors: Erizal Sugiono, Andi Wijaya, Anwar Santoso, Ferry Sandra, Ilham Jaya Patellongi, Irawan Yusuf

BACKGROUND: The association between caffeine with blood pressure (BP) still remains controversial. Caffeine is mainly metabolized by cytochrome-P450 (CYP)1A2 enzyme. Polymorphism of CYP1A2 is known to cause interindividual variation on enzymatic activity, thus affects caffeine metabolism and its effect on cardiovascular (CV) system. METHODS: We conducted a cross-sectional study and recruited 121 Indonesian subjects aged 25-60 years with varying coffee-drinking habits. DNA was extracted from peripheral blood and genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) technique. Blood pressures were measured in the morning prior to the daily activity. Caffeine concentration in blood plasma was measured using high-performance liquid chromatography (HPLC). The differences between variables were analyzed using MannWhitney analysis and the correlations among variables were determined using multivariate logistic regression analysis. RESULTS: This study showed that the frequencies of single nucleotide polymorphisms (SNPs) among Indonesian were 31.8%, 18.2%, 25.2% and 24.8% respectively for CYP1A2*1A, CYP1A2*1B, CYP1A2*1C and CYP1A2*1F alleles. The genotype analysis showed that the subject number of the wild type (*1A/*1A) and the variants were 9.92% and 90.08%. There were no significant differences in term of BP levels among CYP1A2 genotypes and coffee drinking habit groups. CONCLUSIONS: The frequencies of CYP1A2 SNPs in Indonesian are different with frequencies in other populations. Since the association were not statistically significant, CYP1A2 polymorphisms as the predictor of elevated blood pressuare should be investigated further.

84-103

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: The strong inverse association of plasma levels of high-density lipoprotein cholesterol (HDL-C) with coronary heart disease (CHD) found in human epidemiological studies led to the development of the ‘HDL-C hypothesis’, which posits that intervention to raise HDL-C will result in reduced risk of CHD. However, recent evidence has raised doubts about the hypotheses that elevating HDL-C is necessarily therapeutic. Genetic variations that associate with altered HDL-C do not strongly associate with altered cardiovascular disease risk. CONTENT: HDL-mediated cholesterol efflux from macrophage foam cells or measurements of the flux of cholesterol from macrophages to the liver and feces seem to correlate better with atherosclerotic burden than with HDL-C levels. Thus, it may be time to modify the HDL-C hypothesis to the ‘HDL flux hypothesis’, where intervention to promote cholesterol efflux and reverse cholesterol transport will reduce CHD risk, regardless of whether it affects plasma HDL-C levels. A deeper understanding of the complex biology of HDL metabolism and its relationship to reverse cholesterol transport and atherothrombotic events is urgently needed. This might lead to biomarkers of HDL flux and functionality that are more informative than simple measurements of HDL-C levels. SUMMARY: It is now clear from recent clinical trial and genetic studies that some approaches to raising HDL-C levels may have no effect on CHD. This suggests the need to evaluate HDL-C-raising therapies in different clinical populations, as well as therapies targeted toward HDL flux and function rather than simply HDL-C elevation. Perhaps moving from a focus on the HDL-C hypothesis to a focus on the HDL flux hypothesis will permit a biologically based reassessment of the optimal therapeutic approach to targeting HDL for reduction in cardiovascular risk.

123-7

Authors: Erika Diana Risanti, Aditya Kurniawan, Laila Wahyuningsih, Ery Kus Dwianingsih, Hanggoro Tri Rinonce, Jajah Fachiroh

BACKGROUND: Hypermethylation of RASSF1A and CDKN2A is one of epigenetic factor underlies nasopharyngeal carcinoma (NPC) development. Smoking behavior as an NPC’s risk factor causes aberrant DNA methylation. RASSF1A and CDKN2A promoter hypermethylation from peripheral blood cells correlates with smoking behavior. The use of body fluids including peripheral blood as a specimen for DNA methylation analyzes are widely developed, as less invasive method compared to the use of tissue biopsy. This study aims to observe the association between RASSF1A and CDKN2A methylation in peripheral blood and smoking behavior among NPC patients. METHODS: Newly diagnosed NPC subjects were recruited from ear-nose-throat (ENT) outpatient clinic of Dr. Sardjito Hospital, Yogyakarta. DNA from buffycoat of 19 smokers and 20 non-smokers NPC’s patients were isolated. Bisulphite modification was applied to 500 ng of the isolated DNA. The methylation status was detected by MSP (methylation-specific polymerase chain reaction (PCR)). The association between smoking status and promoter hypermethylation was analysis using Chi-Square test. RESULTS: MSP analysis of RASSF1A showed that 68.42% smoker and 75% non-smoker NPC’s patients were methylated. MSP analysis of CDKN2A showed that 21.05% smoker and 25% non-smoker NPC’s patients were methylated. There was no association between smoking behavior with RASSF1A and CDKN2A methylation (p>0.05). CONCLUSION: Statistical analysis showed that smoking behavior is not associated with methylation of RASSF1A and CDKN2A among NPC’s patients.

133-9

Authors: Aryu Candra Kusumastuti, Martha Ardiaria, Meita Hendrianingtyas

BACKGROUND: Lack of appetite in young children leads to growing incidences of physical and mental growth disorders. Supplementation of certain micronutrients can increase appetite and improve nutritional status. This study aims to analyze the effects of zinc and iron supplementation on appetite, nutritional status and intelligence quotient (IQ) in young children. METHODS: An experimental study withrandomized control group pre/post-test design was conducted in Semarang, Indonesia. A total of 68 children were divided into four groups. The first group was the control group, which was given a placebo; the second group was given a zinc supplement at 10 mg/day; the third group was given an iron supplement at 7.5 mg/day; andthe fourth group was given zinc and iron for three months. Appetite was assessed based on eating frequency and energy intake. Nutritional status was assessed by weight per age (W/A) and height per age (H/A) z score. IQ score was assessed based on Wechsler Preschool and Primary Scale of Intelligence (WPPSI). RESULTS: Before intervention, low zinc intake was observed in 27.7% of the subjects and low iron intake was observed in 58.5% of them. After intervention, appetite in the second and fourth groups increased. W/A z score increased in the second and third groups. IQ score increased in the third group. No significant effect on H/A z score was observed in all groups. CONCLUSION: Supplementation of zinc and iron for three months had a positive effect on appetite, body weight and IQ score but no significant effect on body height.

144-7

Authors: Parawansah, Tomy Nurtamin, Sufiah Asri Mulyawati, Nuralifah Nuralifah, Wa Ode Arlina Misnaeni

BACKGROUND: The purpose of this research is to understand the secondary metabolites of Momordica charantia L. extract, as well as to disclose the potential of M. charantia extract in the phagocytic activity and capacity of peritoneum macrophages. METHODS: Examination of immunomodulatory effect was done by giving M. charantia ethanol extract on 5 treatment groups, given intra-peritoneally to mice daily. Echinacea extract as positive control and double distilled water as negative control were also given. On the 8th day, mice were infected with Staphylococcus epidermidis. After 30 minutes, peritoneum fluid was obtained to observe the activity and capacity of macrophage cells. RESULTS: The results showed significant phagocytic activity (p<0.05) at a concentration of 1,200 ppm compare to the other groups. Meanwhile the macrophage cell capacity was found statistically insignificant (p>0.05). The highest phagocytic activity was the group treated with 1,200 ppm (62%), significantly higher than other groups. CONCLUSION: The secondary metabolite content of M. charantia is alkaloids, flavonoids, tannins, saponins, and triterpenoids. The 1,200 ppm M. charantia ethanol extract is potential in inducing phagocytic activity and capacity. These results indicate that the M. charantia can be suggested as a natural immunomodulator.

156-64

Authors: Adelina Simamora, Lusia Paramita, Nur Azreen, Adit Widodo Santoso, Kris Herawan Timotius

BACKGROUND: The leaf and fruit of Psidium guajava L. are potential for neutraceutical beverage especially for antidiabetic drink. The aims of this study were to determine the antidiabetic activity of aqueous extract of leaf (LE) and fruit (FE) from P. guajava. METHODS: Both extracts were investigated for their inhibitory effect on α-glucosidase activity in vitro. Their antioxidant activities were measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, ferrous ion chelating, reducing power and phosphomolybdate methods. RESULTS: The IC50 of LE, FEandacarbose as a positive control were 5.67, 428.00 and 823.99 μg/mL, respectively. The enzyme kinetic analysis indicated that LE inhibited α-glucosidase in a competitive inhibition type, similar to that of acarbose. Both extracts showed antioxidant activities, with LE showed stronger activities than FE in all methods. In DPPH method, IC50 of LE and FE were 74.77 and 843.84 μg/mL respectively, compared to 53.24 and 21.36 μg/mL for reference antioxidants butylated hydroxytoluene (BHT) and ascorbic acid (AA), respectively. In ferrous ion chelating activity, the IC50 were 147.07 and 2105.05 μg/mL for LE and FE, whereas ethylenediaminetetraacetic acid (EDTA) as a control sample was 66.50 μg/mL. In reducing power and phosphomolybdate methods, at different concentrations, the activities of LE, FE, and standard compounds showed the following order: AA > BHT > LE > FE. CONCLUSION: LE from P. guajava exhibited excellent inhibitory activity against α-glucosidase. In addition, LE had better antioxidant acivities than FE. This study can recommend the aqueous extract from P. guajava as a promising candidate for neutraceutical drink for prediabetic and diabetic patients.

171-8

Authors: Ahmad Mozafar, Davood Mehrabani, Akbar Vahdati, Ebrahim Hosseini, Mohsen Forouzanfar

BACKGROUND: Stem cell-based therapy is one of the newest and evolving techniques in reproductive medicine. The aim of this study was to investigate the effect of allogeneic bone marrow mesenchymal stem cells (BM-MSCs) transplantation on the testis of busulfan induced azoospermia in Balb/C mice. METHODS: Eighteen adult Balb/C mice were divided into three equal groups including control, busulfan and busulfan+cell therapy (busul+CT). For induction of azoospermia, busulfan and busul+CT groups received two injections of 10 mg/Kg of busulfan intraperitoneally with 21 days interval. In the cell therapy group 35 days after the last injection of busulfan, cluster of differentiation (CD)90+/CD34-/CD45- BM-MSCs were injected into the efferent duct of testis. Eight weeks after the BM-MSCs therapy, mice were sacrificed and tissues were taken for histological and histomorphometric evaluations. RESULTS: In busul+CT group, cellular and total diameters and cellular and cross-sectional areas significantly increased in comparison to busulfan group (p˂0.001), but there were no significant differences between busul+CT and control group (p˃0.05). Numerical density and tubular count per area unit in busul+CT and control groups were significantly less than busulfan group (p˂0.001), but there were no significant difference between busul+CT and control group (p˃0.05). The luminal diameter and area showed no significant change in all groups (p˃0.05). In busul+CT group, spermatogenesis index significantly increased when compared to busulfan and control groups (p˂0.001 and p˂0.05, respectively). CONCLOSION: Histomorphometric findings showed CD90+/CD34-/CD45- BM-MSCs transplantation on the testis of busulfan-induced azoospermic in Balb/C mice recovered spermatogenesis.

184-91

Authors: Mochammad Thaha, Maulana Antiyan Empitu, Ika Nindya Kadariswantiningsih, Cahyo Wibisono Nugroho, Nurina Hasanatuludhhiyah, Haerani Rasyid, Zaky El Hakim, Maulana Muhtadin Suryansyah, Rieza Rizqi Alda, Mohammad Yusuf Alsagaff, Mochammad Amin, Djoko Santoso, Yusuke Suzuki

BACKGROUND: Obesity is an important cardiovascular risk factor and associated with low grade inflammation in chronic kidney disease (CKD) patients. This study aims to assess the association between body fat with serum high sensitivity C-reactive protein (hs-CRP) level in CKD patients. METHODS: A cross-sectional study was performed in 71 CKD patients. Anthropometric measurements included body weight, height, body mass index (BMI), body fat percentage (BFP), skinfold thickness (SKF) of triceps and biceps were performed by trained physician. BFP was calculated using Kwok’s Formula and hs-CRP was measured by Particle enhanced Turbidimetry. RESULTS: The averaged BMI of our subjects was 25.8±4.4. There was no significant difference in BMI between pre-dialysis and hemodialysis CKD patients. Positive correlation was found between BFP and hs-CRP (r=0.266; p<0.05), while there was no significant correlation between BMI and hs-CRP. CONCLUSION: Body fat percentage was associated with hs-CRP. Hence, it will be more beneficial to assess nutritional status in CKD using BFP rather than BMI alone since it was demonstrated to correlate with hs-CRP in our study.

1-15

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Low back pain (LBP) mostly caused by disc degeneration, reflects to a tremendous of health care system and economy. More knowledge about these underlying pathologies will improve the opportunities that may represent critical therapeutic targets. CONTENT: Basic research is advancing the understanding of the pathogenesis and management of LBP at the molecular and genetic levels. Cytokines such as matrix metalloproteinases, phospholipase A2, nitric oxide, and tumor necrosis factor-α are thought to contribute to the development of LBP. Mesenchymal stem cells (MSCs) transplant to cartilage-like cells and secrete extracellular matrix and encourage nucleus pulposus (NP) cell activity inhibiting NP cell apoptosis, together with some chemical mediators such as cytokines and growth factors become a safe and effective new strategy for intervertebral disc degeneration (IDD) treatment and regeneration. SUMMARY: IDD occurs where there is a loss of homeostatic balance with a predominantly catabolic metabolic profile. A basic understanding of the molecular changes occurring in the degenerating disc is important for practicing clinicians to help them to inform patients to alter lifestyle choices, identify beneficial or harmful supplements, or offer new biologic, genetic, or stem cell therapies.

35-9

Authors: Ferry Sandra

BACKGROUND: Generally ameloblastoma is a locally aggressive, slow growing, non-metastatic epithelial odontogenic benign tumor. However, rarely some ameloblastoma can metastasize in spite of a benign histologic appearance. Targeting ameloblastoma by inducing it into apoptosis could be a beneficial strategy, since many ameloblastoma cases were reported recurrent after surgical therapy. CONTENT: To investigate ameloblastoma in cellular aspect,cytological pattern of ameloblastoma was divided intoouter layer/peripheral and inner layer/central cells. Tumor necrosis factor (TNF)-α, Fas ligand (FasL), TNF receptor (TNFR)1/death receptor (DR)1, TNFR2/DR2, DR4, DR5andFas were highly expressed in central than peripheral cells. Despite inducing apoptosis, TNF-α can induce PI3K leading to Akt and p44/42 mitogen-activated protein kinases (MAPK) activation in AM-1 cells, which later induce cell survival and proliferation. Therefore apoptotic induction in ameloblastoma should be suggested in higher TNF-α concentration. Expression of FasL and Fas are closely associated with squamous metaplasia and granular transformation of the tumor cells, suggesting that apoptosis induced by FasL may play a role in the terminally differentiated or degenerative ameloblastoma cells. TNF-related apoptosis-inducing ligand (TRAIL) has emerged as an apoptotic inducing anticancer agent in tumor cells specifically. TRAIL induced activation of caspases, lowering mitochondrial membrane potential, high number of apoptotic cells in ameloblastoma cells. Therefore, TRAIL could be a potential agent for targeting ameloblastoma, although further study should be explored. SUMMARY: Targeting ameloblastoma by inducing it into apoptosis could be achieved effectively, although some criteria should be considered. Therefore understanding the underlying apoptosis signaling pathways are necessary for inducing ameloblasotma into apoptosis. Investigations on other apoptosis-related molecules, potential apoptosis-inducing natural products, and novel approach in reprogramming, are important in the future for a better anagement of ameloblastoma.

46-50

Authors: Hartanto Bayuaji, Heda Melinda Nazaruddin Nataprawira, Herri Suhari Sastramihardja

BACKGROUND: Polycystic ovary syndrome (PCOS), a common reproductive endocrinologic disorder in woman, was considered to be related to sleep disturbance. This study is aimed to analyze the correlation between excess androgen markers and Athens Insomnia Scale (AIS) in PCOS. METHODS: This observational, cross-sectional study of PCOS was conducted to 31 subjects to evaluate the correlation between serum total testosterone, sex hormone binding globulin (SHBG) and free androgen index with the incidence of sleep disturbance using AIS. Sleep disturbance was present if the score ≥6. The correlation between excess androgen markers and sleep disturbance was analyzed using Pearson’s coefficient of correlation or Spearman’s rho test. Correlation coefficient more than 0.5 with p<0.05 was considered significant. RESULTS: Out of 31 PCOS subjects aged 20-40 years, 39% subjects had AIS score ≥6. Mean serum testosterone in AIS score >6 group was higher than AIS score <6 group but not statistically significant (46.68 vs. 28.49 ng/mL, p>0.05). No significant correlation was found between serum total testosterone, SHBG and free androgen index with AIS score. After adjusting for AIS score, there was the moderate positive correlation between serum total testosterone level although not statistically significant (r=0.54, p=0.07). CONCLUSION: The serum total testosterone level might influence the occurrence of sleep disturbance in PCOS.

56-61

Authors: Edhyana Sahiratmadja, Simeon Penggoam, Ani Melani Maskoen, Alvinsyah Adhityo Pramono, Dias Aryani, Nurul Setia Rahayu, Ramdan Panigoro

BACKGROUND: N-acetyltransferase-2 (NAT2) enzyme, encoded by NAT2 gene, plays a key role in metabolism of anti-tuberculosis (TB) drug isoniazid. Polymorphisms in NAT2 gene may result in different responses to TB therapy. Since TB prevalence in the eastern part of Indonesia is high, the aim of this study is to explore the distribution of NAT2 gene polymorphisms among population from Kupang, Nusa Tenggara Timur. METHODS: A total of 234 respondents were included from Kupang in 2012. Polymorphisms of NAT2 gene were examined using mass screening platform and the genotypes distribution were presented in percentage. To confirm NAT2 gene polymorphisms, polymerase chain reaction (PCR)-sequencing was performed in a subset of population. RESULTS: The polymorphisms of NAT2 gene showed that the distribution of rs1801279 for GG genotype was 100%; whereas the genotype distribution of rs1799930 for GG, GA and AA was 57%, 35.1% and 7.9%, respectively. In a subset of individuals (n13), acetylator status was well determined by PCR-sequencing, resulting in individual with wild type fast acetylator (NAT2*4; n4), intermediate (NAT2*4/*5 or NAT2*4/*6 or NAT2*4/*7; n7) and poor acetylators (NAT2*6/*6 or NAT2*7/*7; n2). CONCLUSION: The amino acid change in rs1799930 result in intermediate and poor acetylator status in Kupang population. This may lead to suboptimal response of TB therapy. Assessing acetylator status before TB therapy is important and may serve as personalized INH therapy.

66-73

Authors: Dwi Ramadhani, Siti Nurhayati, Tur Rahardjo, Mukh Syaifudin

BACKGROUND: Mamuju area in West Sulawesi considered as the high natural background radiation area in Indonesia. Our previous study showed that the mean mitotic index (MI) and nuclear division index (NDI) in lymphocytes of Botteng Village, Mamuju inhabitants was lower compared to control samples. To validate our previous study results, here in this study the evaluation of cell proliferation markers which were MI and NDI in peripheral blood lymphocytes (PBL) of Takandeang Village inhabitants was conducted. METHODS: A total 60 people were enrolled in this study, consisted of 35 samples from Takandeang Sub-Village and 25 from normal background radiation area. MI was calculated manually and automatically using Metafer 3.11.2 imaging system. The NDI defined as proportion of mononucleated, binucleated, trinucleated and tetranucleated cells were conducted using cytokinesis block micronucleus (CBMN) assay. RESULTS: The results of this study showed that the mean manual MI in Takandeang Sub-Village inhabitants was lower compared to control group (4.96±2.25 vs. 5.93±2.14). In contrast, the mean automatic MI (20.37±10.49 vs. 18.87±7.49) and NDI (1.555±0.174 vs. 1.523±0.112) in Takandeang Sub-Village inhabitants was higher compared to the control group. Statistical analysis revealed that the difference of mean manual MI, automatic MI and NDI in Takandeang Sub-Village inhabitants was not significantly different compared to the control group (p>0.05). CONCLUSION: It can be concluded that based on this study the chronic low radiation dose exposure in Takandeang Sub-Village, Mamuju has no significant effect on the lymphocytes proliferation.

79-83

Authors: Hartono Apriliasta Purba, Santi Syafril, Dharma Lindarto

BACKGROUND: Inflammation is one of the factors that associated with insulin resistance which causes type 2 diabetes mellitus (T2DM). Inflammation can be measured by using high sensitivity C-reactive protein (hs-CRP), a quantitative measurement of CRP concentration. Puguntano is a plant that traditionally used for antidiabetic treatment found in North Sumatera. The objective of this study is to know the effect of Puguntano extract on hs-CRP level in newly diagnosed T2DM patient. METHODS: The study was a randomized controlled clinical trial on 24 newly diagnosed T2DM patients. Subjects were divided into 2 groups. The first group was given Puguntano extract with starting doses at 100 mg daily, then the dose was increased in titration up to a maximum dose of 2x100 mg daily depending on blood sugar level monitoring. Meanwhile, the other group (which serves as a control group) was given metformin with starting doses at 500 mg daily, then the dose was increased in titration up to a maximum dose of 3x500 mg per day depending on blood sugar level monitoring. The hs-CRP and HbA1c are assessed at the baseline and after 12 weeks of treatment. RESULTS: The hs-CRP level was decreased in Puguntano group by 1.41 (-7.54-0.52) (p=0.06) and decrease by 0.58 (-7.86-33.41) (p=0.695) in metformin group. There was no difference in hs-CRP level between the two groups (p=0.630). CONCLUSION: Puguntano extract showed the effect of lowering HbA1C and hs-CRP level in T2DM patients, although it was not statistically significant.