The Indonesian Biomedical Journal

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.