The Indonesian Biomedical Journal

1-7

Authors: Inggita Kusumastuty, Frinny Sembiring, Sri Andarini, Dian Handayani

BACKGROUND: Consumption of foods and drinks high in energy, fat, and/or sugar beyond the recommended quantities can cause obesity, which triggers the incidence of brain nerve cell death related to oxidative stress, high levels of tumor necrosis factor (TNF)-α and triglycerides, and low high-density lipoprotein (HDL) levels. Progressive nerve cell death causes decreasing cognitive performance. This study aims to prove that an American Institute of Nutrition committee in 1993 (AIN-93M) diet modified with high-fat-high-fructose (HFHF) can decrease the number of hippocampal neurons. A decrease in the number of hippocampal neurons indicates progressive nerve cell death. METHODS: An experimental study using a post-test control group design was carried out using male Sprague Dawley rats. Samples were selected using simple random sampling to divide them into two groups, Group I was AIN-93M-modified HFHF diet (n=14) and Group II was AIN-93M standard (n=16). The number of visible neurons was measured in the hippocampus area of Sprague Dawley rats’ brains, stained with haemotoxylin and eosin (H&E) and scanned under 400x magnification. Neurons were counted in 10 visual fields using the "Cell_Count" application. RESULTS: The data were analysed by Pearson’s correlation test using SPSS. The results show that rats in Group I had a greater weight gain and fewer neurons than those in the Group II (p=0.023, r=-0.413). CONCLUSION: The consumption of foods high in fat and fructose can cause an increase in nerve cell death, as shown by the decrease in the number of hippocampal neurons.

15-8

Authors: Hudila Rifa Karmia, Afriwardi Afriwardi, Hirowati Ali, Johanes Cornelius Mose, Yusrawati Yusrawati

BACKGROUND: Based on 'vascular disorder of pregnancy' terminology, preeclampsia primarily was not a hypertensive disorder, but a vascular disorder (general vasospasm) in pregnancy due to idiopathic etiology. The overall incidence of preeclampsia was 5-14% of all pregnancies. One of the substances responsible for regulating vascular tone is nitric oxide (NO), which produced in endothelial blood vessels. NO and L-citrulline are produced altogether by the reaction between L-arginine and oxygen. L-citrulline levels reflected NO production. This study was aimed to assess the correlation between the L-citrulline level and blood pressure in severe preeclampsia. METHODS: This cross-sectional study was done in Dr. M. Djamil Central General Hospital, Padang, Indonesia The sample size was 36 samples of pregnant women with severe preeclampsia and had yet been given antihypertensive therapy. Sampling was done by consecutive sampling from Obstetrics and Gynecology Division. After maternal examination and measurements L-citrulline levels of cubital venous blood by using enzyme-linked immunosorbent assay (ELISA) method, Pearson correlation was performed to assess the relationship between variables for normally distributed data and Spearman's correlation for abnormal distribution data with significance level p<0.05. RESULTS: Means of L-citrulline levels, systolic blood pressure, diastolic blood pressure, and arterial pressure (MAP) were 87.21 nmol/mL, 179.4 mmHg, 108.3 mmHg, and 132.1 mmHg, respectively. Correlation of the L-citrulline level with systolic blood pressure, diastolic blood pressure, and MAP were -0.08, -0.175, and -0.136 (p>0.05), respectively. CONCLUSION: L-citrulline levels had no correlation with blood pressure in severe preeclampsia.

27-33

Authors: Fitriyadi Kusuma, Andrijono Andrijono, Ani Retno Prijanti, Laila Nuranna, Sri Mutya Sekarutami, Bambang Sutrisna, Ferry Sandra

BACKGROUND: Cervical cancer is one of the most prevalent cancers in women. Even with similar clinicopathologic features, radiotherapy outcomes are still vary among patients. This research was conducted to measure radiotherapy responses on cervical cancer patients by using Survivin, Telomerase and Cytochrome C. METHODS: Subjects who matched the criteria were selected and requested to fill questionnaires. Subjects were then evaluated with magnetic resonance imaging (MRI) pre- and post-3D conformal radiotherapy. Histopathological study was conducted using resected tumors to determine the differentiation type. Enzyme-linked Immunosorbent Assays for detection of Survivin, Telomerase and Cytochrome C was performed using the resected tumors as well. RESULTS: There were 76 patients in this study. Mean ofage was 50 and diameter of tumor size was 5.35 cm. Mean levels of Survivin, Telomerase and Cytochrome C were 632.82 pg/mL, 5.59 pg/mL and 204.75 ng/mL, respectively. There were significant correlations between radiotherapy response and Survivin (p=0.041) or Telomerase (p=0.022). Subjects with lower Survivin level (<932 pg/mL) had higher 1-year survival rate (63%) than subjects with higher Survivin level (50%). Similar results were obtained for subjects with lower Telomerase level (<5.75 pg/mL), who had higher 1-year survival rate (60%) than subjects with higher Telomerase level (43%). CONCLUSION: Since radiotherapy response is significantly correlated with Survivin and Telomerase levels and subjects with lower Survivin or Telomerase level have higher 1-year survival rate, it can be suggested that Survivin and Telomerase could be potential predictors of radiotherapeutic response for subjects with stage IIIB cervical squamous cell carcinoma.

40-4

Authors: Ismawati Ismawati, Enikarmila Asni, Mukhyarjon Mukhyarjon, Ilhami Romus

BACKGROUND: Diabetes mellitus (DM) is associated with an accelerated atherosclerotic macrovascular disease affecting medium-sized arteries. Several evidences support the role of oxidative stress in atherogenesis. However, the role of alpha lipoic acid (ALA) to prevent atherosclerosis is still debatable. This study was conducted to determine the effect of 60 mg/kg/day ALA for 21 days toward the expression of intercellular adhesion molecule-1 (ICAM-1) in rat model. METHODS: Eighteen male rats were divided into three groups labelled as control group, type 2 DM (T2DM) group, and T2DM+ALA group. The T2DM rat models were created by intraperitoneally injecting 50 mg/kg streptozotocin, followed by 110 mg/kg nicotinamide. Immunohistochemistry was used to evaluate the ICAM- 1 expression in rats. Quantitative image analysis of immunohistochemical stains was done on the abdominal aorta using Adobe Photoshop CS3 to find the area percentage and intensity. Kruskal-Wallis and Mann-Whitney tests were used to compare the mean value of area percentage and intensity. RESULTS: There was an increase in area percentage and intensity of ICAM-1 expression. The highest area percentage of ICAM-1 expression was found in the DM group, while the lowest was found in the control group. There were significant differences in the area percentage and intensity between DM+ALA group and DM group, where the area percentage and intensity of ICAM-I in DM group was higher than the DM+ALA group. CONCLUSION: In conclusion, our results demonstrate that ALA inhibits the expression of ICAM-1 in T2DM rat models.

51-6

Authors: I Gede Widhiantara, Anak Agung Ayu Putri Permatasari, I Wayan Rosiana, I Wayan Putu Sutirtayasa, Ferbian Milas Siswanto

BACKGROUND: Hypoxia has been shown to be able to induce adiposity. However, the mechanism and factors involved in this effect still remains unclear. Hence, we sought to investigate the role of oxygensensitive factors regarding hypoxia-induced adiposity in nematode Caenorhabditis elegans. METHODS: The C. elegans were grown on nematode growth medium (NGM) agar plates seeded with Escherichia coli OP50 at 20°C. The ratio of width/body length was measured using the morphometry analysis. Fat accumulation was examined using Sudan Black methods. Protein levels of sterol binding protein (SBP)-1 were assessed by immunoblotting. Lifespan assay was performed at 20°C and was monitored every two days. RESULTS: The results showed that of all mutant used, only hif-1 mutant which did not experience an increase in the ratio of width/body length (p>0.05) and fat accumulation (p>0.05), indicating that hypoxia-inducible factors (HIF)-1 plays an important role in the pathogenesis of hypoxia-induced adiposity. Both siah-1 and skn-1 mutants experienced SBP-1 protein elevation (p<0.05), and increased fat-6 mRNA expression (p<0.05) which were not experienced by a hif-1 mutant (p>0.05) further supporting that HIF-1 acts as an upstream regulator from SBP-1. CONCLUSION: In general, the results of this study provide evidences of the involvement of the transcription factor HIF-1 in inducing adiposity under the hypoxic conditions. However, we did not find the involvement of seven in absentia homolog-1 (Siah-1) and skinhead-1 (SKN-1).

62-8

Authors: Alit Utamayasa, Mahrus Ahmad Rahman, Teddy Ontoseno, Budiono budiono

BACKGROUND: The angiotensin-converting enzyme inhibitors (ACEIs) have become the forefront of heart failure treatment for more than a decade. Currently, angiotensin receptor blockers (ARBs) are thought to have similar effectiveness. This study aimed to compare the impact of captopril, one of ACEI, and valsartan, one of ARB, on clinical presentation and echocardiographic, electrocardiographic, and chest x-ray improvement in patients with left-to-right shunt congenital heart diseases. METHODS: This study used a double-blind randomized controlled trial of captopril and valsatran to children with left-to-right shunt congenital heart diseases who suffer from heart failure in the Dr. Soetomo General Hospital, Surabaya, Indonesia. Pediatric heart failure scores, echocardiography, electrocardiography (ECG), and chest photographs were examined at the beginning of the study and after 30 days of treatment. RESULTS: A decrease in pediatric heart failure scores were showed after the administration of ACEI (7.06±2.04 vs. 4.75±2.43; p<0.0001; 95% CI: −2.98 - 1.65); ARB (6.81±2.25 vs. 3.94±1.98; p<0.0001; 95% CI: −3.76 to 1.98). The echocardiography examination, an increase in left ventricular end-diastolic volume (LVEDV), left ventricular ejection fraction (LVEF), % fractional shortening (FS), and left ventricular (LV) dimension occurred after the administration of ACEI and ARB. The values also didn’t significantly differrent between the two groups. The ECG evaluation showed a decrease in heart rate frequency after the administration of ACEI (117.75±14.67 vs. 109.63±17.59; p=0.039; 95% CI: −15.78 to −0.46) and ARB (117.10±21.86 vs.108.6±20.66; p=0.006; 95% CI: −14.17 to −2.83). CONCLUSION: ARB showed better outcome in clinical condition, echocardiography, ECG, and chest radiographs.

78-84

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

BACKGROUND: Airway epithelium is the first line of defense against a variety of exposures. Inflammatory processes, hyperresponsiveness and zinc deficiency cause epithelial damage. Zinc is involved in apoptosis and microtubule formation. However, its role in the integrity of bronchial mucosa and cilia is unclear. METHODS: To assess the effect of zinc on the integrity of the bronchial epithelium, 24 male Rattus norvegicus strain Wistar rats were randomized into four experimental groups: normal zinc diet group without zinc supplementation, normal zinc diet group with 60 ppm zinc supplementation, zinc deficient diet group without zinc supplementation, and zinc deficient diet group with 120 ppm zinc supplementation. Bronchial mucosal integrity was measured with the number of epithelial cells, and the number and length of cilia. RESULTS: Number of cell in normal zinc diet group was 8.8±1.82, while it was only 8.1±1.08 in zinc deficient diet group (p<0.001). Number of cilia per cell was 4.6±1.08 in normal zinc diet group, compared to 4.0±0.79 in zinc deficient diet group (p<0.001). Ciliary length also differ by 7.68±0.66 μm in normal zinc diet group and only 5.16±0.91 μm in zinc deficient diet group (p<0.001). CONCLUSION: Zinc supplementation of the normal zinc diet group affected the length of bronchial cilia. Zinc supplementation of the zinc deficient diet group affected the integrity of the bronchial epithelium, which was shown by the number and length of cilia, and the number of epithelial cells.

225-39

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Along with its increased prevalence, in the past decade, cancer had joined the list of chronic debilitating diseases. Nutrition become substantial aspects, due to its time-dependent effect to modulate inflammation thus trigger carcinogenic effects by altering the immune check point. Thus, nutrition contributes to the progression and therapeutic response of cancer, both in human or animal models. CONTENT: Meat is well favored food with appreciable appealing. Due to its high nutritional values it plays a central role in human development. Meat or meat derivate are important sources of proteins, minerals and vitamins. Their nutritional importance is worth compare to their economic impact but recent publication of WHO has set the social alarm about the relationship between red and/or processed meat consumption and cancer. On the other side, some natural or biologic agents may inhibit or reverse tumor growth. Some phytochemical agents including curcumin, resveratrol, lycopene, folates and tea polyphenols clinically proved to tune the signaling pathways regulating cell proliferation and apoptosis in transformed cells, enhance the host immune system and sensitize malignant cells to cytotoxic agents. SUMMARY: Recent studies on chemopreventive agents involves a wide range of molecules, natural (plants, fruits and vegetables) or synthetic will provide better insights for cancer early pathogenesis, important end-point biomarker, and finally potential for reducing the burden of cancer.

247-56

Authors: Nasar Alwahaibi, Siti Balkis Budin, Zariyantey Abdul Hamid, Jamaludin Mohamed, Jalifah Latip, Norzamzurina Binti Ismail, Alefee Bin Ayatillah, Fatmah Ali Matough

BACKGROUND: Tocotrienol-rich fraction (TRF) from palm oil has been shown to exhibit potent antioxidative properties in various pathological conditions. This study aimed to determine the antioxidative properties of TRF on the plasma and erythrocyte membrane lipids composition in streptozotocin-induced diabetic rats. METHODS: Male Sprague-Dawley rats (n=40) were divided randomly into four groups of 10, which were a normal group, a normal group with TRF, a diabetic group, and a diabetic group with TRF. Following four weeks of treatment, lipids compositions in the plasma and erythrocyte membrane were investigated. RESULTS: TRF significantly reduced the plasma total cholesterol, to that in the diabetic group. In addition, supplementation of TRF was able to reduce the very low density lipoprotein cholesterol (VLDL-C), triglycerides levels in treated diabetic rat although no significant different was observed when compared with untreated diabetic rats alone. Furthermore, TRF supplementation showed increased the levels of phosphatidylinositol, phosphatidylserine, and total phospholipid levels in diabetic rats. However, the linoleic acid (18:2) levels were reduced in diabetic rats compared to the control group, and the eicosadienoic acid (20:2) levels were significantly increased by TRF supplementation in diabetic rats. CONCLUSION: Supplementation with 200 mg/kg TRF improves dyslipidemia and that TRF, therefore, might have protective effects against oxidative damage of phospholipids. This study also showed the antioxidative properties of TRF on the erythrocyte membrane lipids composition, in particular, its protective effects against peroxidation of unsaturated fatty acids.

262-6

Authors: Miftakh Nur Rahman, Indriyanti Rafi Sukmawati, Irma Melyani Puspitasari, Miswar Fattah

BACKGROUND: Central obesity is known as a risk factor for type 2 diabetes mellitus (T2DM). Its development is influenced by many factors such as a progressive failure of pancreatic beta cell function. The beta cells increase their function to secret insulin along T2DM development to compensate before it becomes exhausted. Zinc (Zn) plays a crucial role in beta cell function and insulin secretion. The majority of Zn in serum are bound to protein which is not readily available interact with cells. The free Zn in serum has been suggested as being more representative than total Zn in beta cell function. This research aimed to investigate the correlation between serum free Zn and homeostasis model assessment for beta cell function (HOMA-B) and to predict the pancreatic beta cell function in the development of T2DM. METHODS: This study was designed as an observational with a cross-sectional approach. The subjects were centrally obese men aged 30-50 years and who had met the inclusion and exclusion criteria from the screening tests. Control subjects were lean men without T2DM. Serum free Zn and serum total Zn were measured by using inductively coupled plasma-mass spectrometry (ICP-MS). RESULTS: There was positive correlation between serum free Zn and HOMA-B (R=0.361, p-value<0.001) but there was no correlation between serum total Zn and HOMA-B (R=-0.062, p-value=0.563). This study found that if the concentration of serum free Zn >1.7 ug/dL in central obese men was suggested as an excessive function of pancreatic beta cell and as an early warning before its exhausted. CONCLUSION: This study suggested that serum free Zn had a correlation with beta cell function and had a predictive ability for beta cell excessive function before its exhausted.

273-8

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

BACKGROUND: Tobacco smoking plays an important role in nasopharyngeal carcinoma (NPC) risk. The Wingless-related integration site (WNT) Inhibitory Factor 1 (WIF1) as one of the tumor suppressor gene (TSG), functions to control cell proliferation through suppression of WNT signaling pathway. WIF1 methylation is one of epigenetic mechanisms which causes overactivated of WNT pathway in NPC development. WIF1 methylation from buffy coat related to smoking status and NPC risk is not known yet. METHODS: This was a nested case-control study involving 39 newly diagnosed NPC patients and 40 healthy controls. All subjects were enrolled at Dr. Sardjito Hospital, Yogyakarta. Subjects were male, with known smoking status, duration, and quantity of cigarette consumed. Parallel DNA isolated from buffy coat was used for WIF1 DNA methylation analysis, by using methylation-specific PCR (MSP) method. Chi-square analysis was used to determine distribution of differences among groups. Two tailed p-value≤0.05 was considered as statistically significant. RESULTS: No significant difference between WIF1 methylation of cases and controls (p=0.30), nor smoking habit among smokers (p=0.51) and non-smokers (p=0.43). CONCLUSION: WIF1 methylation from buffy coat could not be used as an NPC marker nor as a smoking behavior marker.

286-92

Authors: Irianiwati Widodo, Ery Kus Dwianingsih, Teguh Aryandono, Soeripto Soeripto

BACKGROUND: Triple negative breast cancer (TNBC) is a breast cancer subtype with negative expressions of estrogen receptors (ER), progesterone receptors (PR) and Her-2 protein receptors. This subtype exhibits aggressive behavior and poor prognosis since it is unresponsive to hormonal and Her-2 targeted therapy. The identification of clinicopathological characteristics and their prognostic values will provide guidance in developing effective treatment. This study was performed to analyze the clinicopathological characteristics and prognostic significance in Indonesian TNBC. METHODS: Forty paraffin-embedded tissues of TNBC, stage I to IIIA, dating from 2008 to 2010 in Dr. Sardjito Hospital, were enrolled. Survival follow-up was done from January 2008 to June 2013. The samples were immunostained with ER, PR and Her-2 monoclonal antibodies to determine the TNBC subtypes. The clinicopathological characteristics were statistically analyzed. The survival was analyzed using the Kaplan Meier method. The Cox proportional hazards model was used for multivariate analysis. RESULTS: Mean age of TNBC patients was 51.42±11.72 years old, whereas the mean of tumor size was 5.4±2.92 cm. Lymph node metastasis was found in 70% of cases. Number of locally advanced samples (IIIA) was 37.5%, while moderate to high grade samples were 95%. Number of still alive patients at the end of the study was 55%. Number of patients that still alive up to the end of the study was 45.50%. Lymph node was an independent prognostic factor for survival of TNBC patients as positive status of lymph node increases the death risk to 6 times higher (p=0.02, RR=6.6). However, after being adjusted with age and stage, lymph node status was suggested as a modifier effect for the survival of TNBCs patients (RR=0.19). CONCLUSION: Indonesian TNBC patients were mostly found already with large tumor size, lymph node metastasis, high pathological grade and relatively young age. Lymph node status serves as a modifier effect for the survival of Indonesian TNBC.

299-303

Authors: Muhammad Diah, Rahmawati Rahmawati, Aznan Lelo, Zulfikri Muhktar, Dharma Lindarto, Ferry Sandra

BACKGROUND: Tumor necrosis factor (TNF)-α, an important primary pro-inflammatory cytokine, has a crucial role in the pathogenesis of atherosclerosis. Since the pathophysiological mechanism of coronary slow flow (CSF) is not fully understood, we investigated the level of TNF-α in coronary artery disease (CAD), CSF and healthy subjects. METHODS: This study was conducted in cross-sectional design involving 16 CAD, 18 CSF and 18 healthy subjects. Coronary angiography was recorded at the left anterior oblique, cranial, right anterior oblique, caudal, and horizontal positions. The flow in coronary arteries of the subjects were assessed using Thrombolysis in the Myocardial Infarction (TIMI) frame count method. Peripheral blood-derived serum was collected and level of TNF-α was determined by using highly sensitive enzymelinked immunosorbent assay (ELISA). RESULTS: No significant difference in level of TNF-α in CAD, CSF and healthy subjects (2.72±2.64 pg/mL, 1.88±0.8 pg/mL, 1.64±0.35 pg/mL, respectively) (p=0.087). In addition, there was no correlation between the concentration of TNF-α and TIMI frame count (r<0.2, p>0.05). CONCLUSION: There was no significant difference of TNF-α level in CAD, CSF and healthy subjects. In addition, there was no correlation between the TNF-α level with TIMI frame count as well. Nevertheless, further clinical studies with more subjects are needed.

314-9

Authors: Rininta Firdaus, Hege Hartz Jartun, Miriam Khider

BACKGROUND: Rapid and accurate identification of Staphylococcus capitis is required to provide a better prognosis for endocarditis patients and tackle the emergence of multidrug resistant strains of the bacteria in hospitals. The current study was aimed to develop polymerase chain reaction (PCR) assay for specific identification of S. capitis using sodA and gap genes as markers. METHODS: Five sequences of sodA and sixteen sequences of gap registered in GeneBank were analysed using bioinformatic tools. PCR primers were designed based on the conserved and specific regions of sodA and gap. Four clinical isolates of S. capitis (named no. 56-59) and six reference strains of coagulase-negative staphylococci (CoNS) species including S. epidermidis ATCC 35984, S. epidermidis 48951I/09, S. lugdunensis 44987/09, S. sciuri 109645I/08, S. warneri 135612/09, S. hominis 114202/08 were used to validate the conventional PCR system. RESULTS: The current PCR system only amplified the DNA template of S. capitis. Current primers specifically targeted S. capitis as the agarose images only showed bands from S. capitis samples. CONCLUSION: The sodA and gap genes might serve as effective markers for identification of S. capitis using conventional PCR. The PCR assay in the current study was able to identify five clinical isolates of S. capitis accurately without mispriming, misamplification and misidentification. The PCR system was also able to discriminate other CoNS including S. epidermidis, S. lugdunensis, S. sciuri, S. warneri and S. hominis.

327-37

Authors: Adnan Jassim Mohammed Al-Fartosy, Nadhum Abdulnabi Awad, Rsha Abdulradha Mahmood

BACKGROUND: To date, there is no study regarding women with unexplained infertility in the province of Basrah, Iraq. This study was conducted to compare the level of leptin, oxidant/antioxidant status, and some trace element levels in women of healthy control and unexplained infertility. METHODS: From total 96 women subjects, 32 subjects had primary unexplained infertility, 31 had secondary unexplained infertility, while 33 fertile women were taken as controls. The body mass index (BMI) of subjects was calculated with standard procedures. The level of fasting serum leptin, luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (PRL), thyroid-stimulating hormone (TSH), 4-hydroxynonenal (4-HNE), lipid hydroperoxide (LOOH), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and the total antioxidant capacity (TAC) were determined by enzyme-linked immunosorbent assay (ELISA) methods. Magnesium (Mg) and zinc (Zn) were measured by flame, copper (Cu) by flameless atomic absorption spectrometry, while Selenium (Se) in whole blood was determined using hydride generation method. RESULTS: Non-significant changes (p>0.05) were seen in the level of BMI, leptin, TSH, FSH, LH and LH/FSH ratio. Level of 4-HNE, LOOH and Cu were significantly increased (p<0.01), while level of GSH, SOD, CAT, TAC, Se, Zn and Mg were significantly decreased (p<0.01) in primary and secondary type of unexplained infertility, compared to control. CONCLUSION: Serum leptin hormone is significantly associated with BMI and physical activity in both groups. The significant difference in oxidant/antioxidants status between unexplained infertile and fertile women may suggests a possible additional role of oxidative stress mechanisms in pathophysiology of unexplained infertility.

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.