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INDONESIA
The Indonesian Biomedical Journal
ISSN : -     EISSN : -     DOI : -
Core Subject : Health, Science,
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Articles 621 Documents
Obtaining Specific Hybridomas for Ki-67 Protein Immunodetection Aigerim Turgimbayeva; Sailau Abeldenov; Nurgul Sarina; Bekbolat Khassenov; Saule Eskendirova
The Indonesian Biomedical Journal Vol 13, No 3 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i3.1531

Abstract

BACKGROUND: Active proliferation is specific property of a tumor cells. However, the cost of the analysis is high due to commercial anti-Ki-67 mAbs used as the main immunoreagent for reliable identification of proliferating cells. In this study, recombinant protein was used to obtain specific mAbs for Ki-67 biomarker immunodetection.METHODS: Codon optimized fragment of ki-67 gene was cloned into the pET28c(+)vector. The recombinant protein was purified by immobilized metal affinity chromatography (IMAC) and confirmed by liquid chromatography–mass spectrometry (LC-MS)/MS. Hybridoma cells were obtained by fusing myeloma cells with mouse spleen cells immunized with recombinant antigen. The specificity and activity of mAbs was determined by enzyme-linked immunosorbent assay (ELISA), Western blot and immunocytochemistry.RESULTS: The pET-28c(+)/ki-67 plasmid, which encodes 355 amino acid protein, was obtained. Analysis by LC-MS/MS of the recombinant antigen showed that 77.5% of the amino-acid sequence belonged to Ki-67 protein. Recombinant fragment of Ki-67 protein was used to obtain specific hybridoma strains. ELISA and Western blot demonstrated high affinity and the specificity of obtained mAbs against Ki-67 protein. Newly generated anti-Ki67 mAbs detected target protein in proliferating cells of MCF-7 cell line by immunocytochemistry.CONCLUSION: Newly developed mAbs are potentially useful as an immunodiagnostic tool for assessing the proliferative activity of breast tumor cells using immunocytochemistry.KEYWORDS: breast cancer, Ki-67, monoclonal antibodies, nuclear antigen, recombinant antigen, tumor cells
Advance-Platelet Rich Fibrin and Hyaluronic Acid Combination Improves Interleukin-6 and Granulation Index in Diabetic Foot Ulcer Patients Ronald Winardi Kartika; Idrus Alwi; Franciscus Dhyanagiri Suyatna; Em Yunir; Sarwono Waspadji; Suzanna Immanuel; Todung Silalahi; Saleha Sungkar; Jusuf Rachmat; Saptawati Bardosono; Mirta Hediyati Reksodiputro
The Indonesian Biomedical Journal Vol 13, No 2 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i2.1501

Abstract

BACKGROUND: Diabetic foot ulcer (DFU) is the most common and threatening complication of Diabetes Mellitus (DM). Ideal wound dressing for DFU management should relieve symptoms, provide wound protection, and encourage healing. Advanced-Platelet Rich Fibrin (A-PRF) and Hyaluronic Acid (HA) have been proven to improve wound healing process. This study was aimed to demonstrate the ability of combination of A-PRF and HA in reducing inflammation and improving DFU tissue healing.METHODS: Twenty DFU subjects were involved in this study, and divided into two groups based on the topical fibrin gel treatment: A-PRF + HA group and A-PRF only group. A-PRF was obtained by peripheral blood centrifugation. A-PRF + HA was prepared by homogening A-PRF and AH with a ratio of 1:0.6. Interleukin-6 (IL-6) level, granulation index (GI), numeric pain score (NPS), and inflammation clinical symptoms (ICS) were assessed on day-0, 3, 7 and 14.RESULTS: Wound swabs’ IL-6 level on day-7 was found to be significantly lower in A-PRF + HA compared to A-PRF alone (p=0.041). The IL-6 level reduction also found to be significant higher either in wound swabs (day 0-7, p=0.015) or fibrin gel (day 0-3, p=0.049; day 0-7, p=0.034). A-PRF + HA treatment significantly increased the GI even since day-3 (p=0.043), with lower NPS (p<0.001), and ICS score.CONCLUSION: The combination of A-PRF and HA increases the GI in DFU healing by reducing the inflammation state which will induce the angiogenesis process, as well as reducing pain in DFU subjects better than A-PRF alone.KEYWORDS: inflammation, interleukin-6, wound healing, angiogenesis, proliferation 
The Potency of NTHi lic1A Gene as a Biomarker in Determining The Severity of Post-Viral Acute Rhinosinusitis Imam Megantara; Ronny Lesmana; Nova Sylviana; Sunarjati Soedigdoadi; Teti Madiadipoera
The Indonesian Biomedical Journal Vol 13, No 3 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i3.1614

Abstract

BACKGROUND: The lic1A gene is an important virulence factor for non-typeable H. influenzae (NTHi), which allows its translocation from the nasopharynx into the sinonasal cavity and modulates more severe inflammatory processes. This study is aimed for identifying the potential correlation between the NTHi lic1A gene expressions and the severity of post-viral acute rhinosinusitis.METHODS: Sixty patients who were diagnosed with post-viral acute rhinosinusitis, were recruited from an ENT clinic in a referral hospital, in Bandung, West Java, Indonesia. All patients underwent a historical assessment and ENT examination. The nasal specimen was taken from the patient’s middle meatal. The NTHi lic1A gene expression was detected using Polymerase Chain Reaction (PCR).RESULTS: We observed that eight patients had the NTHi lic1A (+), with a strong correlation toward the dominant symptoms (nasal obstruction and discharge). In addition, the symptom’s duration of the NTHi lic1A (+) was twice longer than patients with the NTHi lic1A (-). Its severity was significantly more different between the two groups (p=0.034).CONCLUSION: Taken together, the presence of the NTHi lic1A gene is significantly associated with the severity of the disease and the symptom’s duration. Thus, the NTHi lic1A gene could potentially be a good marker for assessing the severity of post-viral acute rhinosinusitis cases in the future.KEYWORDS: H. influenzae, rhinosinusitis, nasal obstruction, virulence factors
Pheretima javanica K. Ethanol Extract Granules’ Effects on Eosinophil Level, Immunoglobulin E Level, and Organs Histopathology in Rattus norvegicus B. Dwi Wahyuni; Joko Waluyo; Jekti Prihatin; Fersty Isna Kusumawardani; Aditya Kurniawan
The Indonesian Biomedical Journal Vol 13, No 2 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i2.1388

Abstract

BACKGROUND: Traditional medicine that might be useful to treat typhoid fever is earthworms (Pheretima javanica K.). It is important to know whether any substances contained in Pheretima javanica K. extract are toxic and have pharmacological effects, which can be observed from the eosinophil and immunoglobulin E (IgE) levels in the blood.METHODS: Thirty Rattus norvegicus B. were included in this study and were separated into 5 groups, with 6 rats in each group. T1, T2, T3, and T4 were groups of rats treated with 50, 300, 2000, and 5000 mg/kg BW of Pheretima javanica K. ethanol extract granules (PJEEG), respectively, while the control group was a group of rats treated with 0.36 mg Ciprofloxacin. The eosinophil number and the IgE concentration were detected. Physical observations were also conducted to detect any allergic symptoms, toxicity symptoms, or histopathological changes.RESULTS: Administration of PJEEG significantly increased the number of eosinophil in every group, from 49.82 cells/μL to 259.26 cells/μL on average (p<0.001). PJEEG did not significantly affect IgE levels (p=0.624). The results showed that the administration did not cause any allergic and toxicity symptoms. Histopathological analysis of rats’ kidneys and liver also showed that the administration had no significant effect on histopathological changes.CONCLUSION: The administration of PJEEG does not cause allergic symptoms, toxicity symptoms, and histopathological changes in rats. The administrations also significantly increase eosinophil number in rats’ blood. However, the concentration of IgE in rats’ blood was not significantly changed after administration of PJEEG.KEYWORDS: allergies, typhoid fever, earthworms, Pheretima javanica K., eosinophil, immunoglobulin E
Assessment of sdLDL-C by Three Different Formula and Its Correlation with Clinical Variables Among Diabetes Individuals with and without Nephropathy Desy Thayyil Menambath; Durga Rao Yella; Ashok Prabhu Khandige; Sudha Kuthethur; Nandini Mangalore
The Indonesian Biomedical Journal Vol 13, No 2 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i2.1312

Abstract

BACKGROUND: Diabetes mellitus is a common disease worldwide which affects renal function. Cardiovascular morbidity and mortality in diabetes patients can be accelerated by dyslipidemia. Small dense lowdensity lipoprotein-cholesterol (sdLDL-C) is atherogenic and its predominance has been known as a cardiovascular risk factor. The study aimed to assess the validity of calculated sdLDL-C using three different formulae and its association with other clinical variables in diabetic patients with and without nephropathy, and also to determine the best suited formula to measure sdLDL-C.METHODS: The study subjects were divided into two groups based on the amount of albumin excreted in the urine. Group I or the control group consisted of diabetic subjects without microalbuminuria, while group II consisted of diabetic subjects with microalbuminuria. Blood glucose, glycated haemoglobin (HbA1c), creatinine, total cholesterol (TC), triglyceride (TG), high-density lipoprotein-cholesterol (HDL-C), LDL-C and apoB were estimated. Three formulae used for the validation of calculated sdLDL-C were TG/HDL, sdLDL (mg/dL) = 0.580 (nonHDL) + 0.407 (direct-LDL-C) – 0.719 (calculated-LDL-C) – 12.05, and LDL-C/LDL apoB.RESULTS: There was no significant difference in sdLDL-C levels of diabetic subjects with and without nephropathy. The sdLDL-C had strong correlation with TC, TG, LDL-C, very-low-density lipoprotein (VLDL), non-HDL and apoB in both study groups. ROC curve showed that LDL-C/LDL apoB derived sdLDL-C had better sensitivity (85%) and specificity (69%) compared to other two measures.CONCLUSION: Though the calculated sdLDL-C do not predict the occurrence of nephropathy in diabetes subjects, it may still be used in conjunction with the traditional markers since it is cost effective. The LDL-C/LDL apoB formula is the best predictor of sdLDL-C among the three equations.KEYWORDS: HDL-C, LDL-C, Microalbuminuria, sdLDL-C, T2DM
Apoptosis and Efferocytosis in Inflammatory Diseases Chandra Agung Purnama; Anna Meiliana; Melisa Intan Barliana; Keri Lestari Dandan; Andi Wijaya
The Indonesian Biomedical Journal Vol 13, No 3 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i3.1608

Abstract

BACKGROUND: Millions of cells in multicellular organisms regenerate every day to replace aged and died cells. Effective cell clearance (efferocytosis) is critical for tissue homeostasis, as the human body recycles its cellular components. We summarize what is known about the mechanisms of efferocytosis and how it impacts the physiology of the organism, effects on inflammation and the adaptive immune response, as well as the consequences of defects in this critical homeostatic mechanism in this review.CONTENT: Cell death is the process by which the human body replaces aged or damaged cells with new ones. It can be triggered by genetically encoded machinery or regulated cell death, or by specific pharmacologic or genetic interventions, resulting in accidental cell death. Dying cells release signals that entice phagocytes to engulf them in a process known as efferocytosis. Efferocytosis is a multistep process involving the release of “find me” and “eat me” signals and destruction of death cells by phagocytes. Different types of cell death including apoptosis and necroptosis can express pro- or anti-inflammatory signals via macrophage activity modulation.SUMMARY: Failed or ineffective efferocytosis can result in disruption of tissue homeostasis, which can contribute to the development of chronic inflammatory diseases such as atherosclerosis, obesity, diabetes, and heart failure. Therefore, any therapeutic strategy that enhances efferocytosis will have a beneficial effect on the treatment of these metabolic disorders.KEYWORDS: apoptosis, necroptosis, phagocytosis, efferocytosis, macrophage.
Curcumin Analogs, PGV-1 and CCA-1.1 Exhibit Anti-migratory Effects and Suppress MMP9 Expression on WiDr Cells Febri Wulandari; Muthi&#039; Ikawati; Mitsunori Kirihata; Jun-Ya Kato; Edy Meiyanto
The Indonesian Biomedical Journal Vol 13, No 3 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i3.1583

Abstract

BACKGROUND: Colon cancer is still a crucial concern in the development of chemotherapeutic drugs due to the drug resistance phenomenon and various side effects to patients. One of the newest compound that show anticancer activities against several cancer cells, Chemoprevention Curcumin Analog 1 (CCA-1.1), has increasingly been explored to overcome the limitation of conventional drugs.METHODS: We evaluated the anti-migratory effect of CCA-1.1 and Pentagamavunone-1 (PGV-1) by using WiDr colon cancer cells. The expression profiles of Tumor Protein 53 (TP53) and Matrix Metalloproteinase-9 (MMP9) in colon cancer were obtained from the UALCAN database. Survival outcomes of TP53 and MMP9 in colon cancer patients were analyzed using the Kaplan-Meier method. We used 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT), scratch wound healing, and gelatin zymography assays to observe the cytotoxic effect, anti-migratory activity, and MMP9 expression, respectively, in CCA-1.1 or PGV-1-treated cells.RESULTS: Level of MMP9 was found significantly overexpressed in the primary tumor and metastasis nodal, while TP53 mutation sample types were observed and influenced the survival outcome in colon cancer patients. CCA-1.1 and PGV-1 exhibited strong cytotoxic activity after 24 and 48 h treatment against WiDr cells. The migration assay demonstrated that PGV-1 and CCA-1.1 at 1 mM inhibited cell migration up to 40% after 48 h in single and combination with doxorubicin. The MMP9 expression was significantly inhibited by 0.5 mM CCA-1.1.CONCLUSION: This study emphasizes that the anti-migratory effect of CCA-1.1 is better than PGV-1 via MMP9 suppression on WiDr. Thus, CCA-1.1 is prominent to be developed as an anti-metastatic agent.KEYWORDS: chemopreventive curcumin analog 1.1 (CCA-1.1), PGV-1, WiDr cells, anti-migration, MMP9
The Performance of Glycated Hemoglobin vs. Oral Glucose Tolerance Test in the Diagnosis of Glycemic Disorders among Women with Polycystic Ovary Syndrome in Southern Iraq Mahmood Thamer Altemimi; Alaa Khattar Musa; Abbas Ali Mansour
The Indonesian Biomedical Journal Vol 13, No 2 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i2.1431

Abstract

BACKGROUND: Obese women with polycystic ovary syndrome (PCOS) are at high risk for developing type 2 diabetes mellitus (T2DM). A baseline oral glucose tolerance test (2-h OGTT) is important to screen for dysglycemia in PCOS particularly those high risk women. Due to its advantages by fasting is not required and less day-to-day variability, glycated hemoglobin (HbA1c) might be a convenient screening tool. This study aimed to evaluate the performance of HbA1c vs. 2-h OGTT in the diagnosis of glycemic disorders in PCOS and to evaluate the correlation between glycemic disorders, insulin resistance (IR), and anthropometric measures.METHODS: One hundred and twenty nine women diagnosed with PCOS according to the Rotterdam 2003 criteria in Basrah were included in the study. All subjects were examined for weight, body mass index (BMI) and waist circumference (WC). Then they were tested for fasting glucose, 2-h OGTT, HbA1c, and fasting insulin to assess IR.RESULTS: The result of 2-h OGTT test showed that there were 21 subjects (16.1%) showed to have IGT. The result of HbA1c test showed that 25 subjects (19.4%) were diagonised with prediabetes. Meanwhile FPG test result showed that 34 subjects (26.4%) were having IFG. The HbA1c examination showed an underdetected the diagnosis of T2DM (0.8%) and an overdeteced diagnosis of prediabetes (19.4%) (p=0.021) and at HbA1c 5.55% (37.2 mmol/mol), the specificity was (74.3%) and sensitivity (56.5%) to discriminate normal from abnormal glucose status. Ninety nine women (76.7%) were either overweight or obese and most of them had IR (76.8%).CONCLUSION: Screening of glycemic disorders is crucial for PCOS by using 2-h OGTT regardless of risk factor and HbA1c seems to be an unsatisfactory screening tool to predict glycemic disorders in women with PCOS.KEYWORDS: PCOS, glycemic disorders, OGTT, HbA1c, insulin resistance, and prediabetes
Programmed Cell Death Protein 1-overexpressed CD8+ T Lymphocytes Play a Role in Increasing Chronic Hepatitis B Disease Progression Fatmawati Fatmawati; Ellyza Nasrul; Nasrul Zubir; Ferry Sandra
The Indonesian Biomedical Journal Vol 13, No 3 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i3.1601

Abstract

BACKGROUND: T lymphocyte activation depends on the balance of co-stimulatory and co-inhibitory signals determined by Cluster of Diffrentiation (CD)28 and Programmed Cell Death Protein 1 (PD-1) expression. Alteration in CD28 and PD-1 expression might affect the progression of chronic hepatitis B (CHB). Current study was conducted to evaluate the correlations of the CD28 and PD-1 expressions of T lymphocytes and CHB progression.METHODS: Subjects were recruited, selected and divided into 3 groups, inactive CHB, active CHB and CHB with End-Stage Liver Disease (ESLD). HBeAg was determined by using Enzyme-Linked Fluorescence Assay while HBV-DNA was carried out by the RT-PCR method. Numbers of T lymphocytes expressing CD3, CD4, CD8, CD45, CD28 and PD-1 molecules were determined by flowcytometry. RESULTS: There was no significant difference in the expression of CD28 by CD4+ and CD8+ T lymphocytes of inactive CHB, active CHB and CHB with ESLD subjects. There was also no significant difference in the expression of PD-1 in CD4+ lymphocytes of inactive CHB, active CHB and ESLD subjects. In contrast there was a significant increase in the expression of PD-1 in CD8+ T lymphocytes of ESLD subjects.CONCLUSION: CD28 expression among CHB subjects was within normal range and not related to disease progression, but PD-1 expression of CD8+ T lymphocyte was increased along with disease progression, especially in CHB subjects with ESLD. This suggests that PD-1-overexpressed CD8+ T lymphocyte play a role in increasing CHB disease progression.KEYWORDS: chronic hepatitis B, CD28, PD-1, T lymphocyte, disease progression
Coffea canephora Bean Extract Induces NIH3T3 Cell Migration Yessy Ariesanti; Ferry Sandra; Bianda Claresta; Livia Alvita
The Indonesian Biomedical Journal Vol 13, No 2 (2021)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v13i2.1522

Abstract

BACKGROUND: Wound healing is an essential biological process that consists of sequential steps aimed at restoring the architecture and function of damaged cells and tissues. There are empirical evidences of using pure coffee bean powder as an alternative medicine in treating various types of wounds. However, there is limited data on coffee-induced wound healing, especially migration of cells. Therefore, current study was conducted to investigate the role of coffee extract in cell migration, especially fibroblast which is important for wound healing.METHODS: Coffea canephora beans were prepared, extracted and added in the NIH3T3 cell culture in final concentration of 2.5% and 5%. Then cytotoxicity test was performed using Na,30-[(phenylamino)-carbonyl]-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzenesulfonic acid hydrate (XTT) assay. Meanwhile, cell migration test was performed with scratch assay. All results were statistically analyzed.RESULTS: The 2.5% or 5% Coffea canephora beans extract (CCBE)-treated NIH3T3 cell numbers were almost similar with the numbers of NIH3T3 cells in starvation medium merely. Meanwhile, 2.5% and 5% CCBE showed significant decrease of the widths of scratched areas compared to starvation medium merely (ANOVA with LSD Post-hoc, p=0.000). After 24 h and 48 h, the average widths of 2.5% and 5% CCBE-treated scratched areas were 235.68±22.79, 50.36±5.29, 229.95±23.01, 27.68±2.83, respectively.CONCLUSION: Since both 2.5% and 5% CCBE are potential in inducing migration of fibroblast (NIH3T3 cell) and do not induce cytotoxicity, the CCBE could be potential as an agent for wound healing.KEYWORDS: coffee, Coffea canephora, NIH3T3, migration, cytotoxicity