The Indonesian Biomedical Journal (Prodia Education and Research Institute)
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Optimization and Validation of a Real Time Reverse Transcriptase Polymerase Chain Reaction with RNA Internal Control to Detect Rubella RNA
Get PDFBACKGROUND: According to a report from WHO, cases of rubella infection in Indonesia has increased up to 10-fold from 2007 to 2011. Despite no data of congenital rubella syndrome in the report, there are approximately 45,000 cases of babies born with heart failure and 0.1-0.3% live births with congenital deafness in Indonesia. Allegedly, rubella infection during pregnancy may play a role in this condition. This study aimed to optimize and validate a real-time reverse transcriptase polymerase chain reaction (RT-qPCR) method to detect rubella virus RNA as an aid for the diagnosis of congenital rubella infection.METHODS: Method optimization was conducted using nucleic acids extracted from Trimovax Merieux vaccine with the High Pure Viral Nucleic Acid Kit. One step RT-qPCR was performed with Quantifast Multiplex RTPCR+R Kit. Target synthetic DNA was designed and used to determine the sensitivity of the method. RNA internal control was synthesized to control the process of extraction and amplification.RESULTS: The analytical sensitivity of this method was as low as 5 copies target synthetic DNA/μl. The mean Coefficient of Variation (CV) % of the critical threshold (Ct) obtained were 2.71%, 1.20%, 1.62%, and 1.59% for within run, between run, between kit lots, and between operators, respectively. Recovery of the target synthetic DNA from amniotic fluid was 100.51% (by the log copies/μl) at the concentration of 1,000,000 copies/μl.CONCLUSION: RT-qPCR is successfully used for the detection of rubella virus RNA in vaccine and synthetic nucleic acid. With its high sensitivity, good precision and recovery, this method offers a means to improve the diagnosis of congenital rubella infection in developing countries like Indonesia.KEYWORDS: congenital rubella, RT-qPCR, prenatal diagnosis, amniotic flui
Caffeic Acid Inhibited Receptor Activator of Nuclear Factor kappaB Ligand (RANKL)-Tumor Necrosis Factor (TNF) alpha-TNF Receptor Associated Factor (TRAF) 6 induced Osteoclastogenesis Pathway
No full textBACKGROUND: Caffeic acid was reported in our previous study to have potential in inhibiting osteoclastogenesis through inhibition of nuclear factor κB (NFκB). Here in our current study, we would like to investigate further the caffeic acid-affected signaling pathway leading to NFκB inhibition. Since tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) plays important role in osteoclastogenesis, we applied TRAF6- transfected RAW264 cells D-Clone (RAW-D) cells as model in this study.METHODS: Caffeic acid in various concentrations was added to in vitro osteoclastogenesis of receptor activator nuclear factor κB ligand (RANKL)-TNFα-induced TRAF6-transfected RAW-D cells. Cells were collected, lysed and immunoblotted to detect TRAF6 expression. To detect tartrate resistant acid phosphatase (TRAP)+ polynucleated cells (PNCs), TRAP staining was performed. Meanwhile, to measure NFκB Activity, cells were transfected with pNFκB-TA-Luc and subjected to Dual Luciferase Reporter Assay System.RESULTS: Caffeic acid did not influence TRAF6 expression of RANKL-TNFα-induced TRAF6-transfected RAW-D cells. Caffeic acid diminished NFκB activity of RANKL-TNFα-induced TRAF6-transfected RAW-D cells in a concentration dependent manner. Significant NFκB activity inhibitions were seen under treatment of 1 and 10 μg/ml caffeic acid. By adding 10 μg/ml caffeic acid in RANKL-TNFα-induced TRAF6-transfected RAW-D cells, TRAP+ PNCs number was significantly suppressed.CONCLUSION: Caffeic acid inhibited RANKL-TNFα-TRAF6-induced osteoclastogenesis pathway. Since caffeic acid did not influence TRAF6 expression, TRAF6-RANK interactions and/or TRAF6 downstream signaling pathway should be further pursued to disclose inhibition mechanism of caffeic acid.KEYWORDS: caffeic acid, osteoclastogenesis, TRAF6, RANKL, TNFα, NFκB, RAW-
Concentration of Endogenous Secretory Receptor for Advanced Glycation End Products and Matrix Gla Protein in Controlled and Uncontrolled Type 2 Diabetes Mellitus Patients
Get PDFBACKGROUND: Advanced glycation end products (AGE) and their receptor (RAGE) system play an important role in the development of diabetic vascular complications. Recently, an endogenous secretory RAGE (esRAGE) has been identified as a novel splice variant, which lacks the transmembrane domain and is secreted in human sera. Interestingly, it was reported that esRAGE binds AGE ligands and neutralizes AGE actions. Many studies have reported that diabetes mellitus correlates with vascular calcification event and increases progressively in uncontrolled diabetes. Matrix Gla Protein (MGP) is known to act as an inhibitor in vascular calcification. The aim of this study was to observe progress of vascular calcification in uncontrolled diabetes patient by biochemical markers MGP as inhibitor in vascular calcification, via mechanism of AGEs.METHODS: This study was an observational study with cross sectional design on adult type 2 diabetic male patients who were defined by the 2011 Indonesian diabetes mellitus consensus criteria.RESULTS: The results of this study showed that there was a positive significant correlation between esRAGE and HbA1C (r=0.651, p=0.009), and negative correlation between MGP and HbA1C (r=-0.465, p=0.081) in controlled diabetes group. In uncontrolled diabetes group there was a positive significant correlation between MGP and HbA1C (r=0.350, p=0.023), despite the fact esRAGE showed no significant correlation with HbA1C. There was no significant difference in level of esRAGE and MGP in controlled and uncontrolled diabetes group, but MGP showed lower level in uncontrolled diabetes group, contrary to esRAGE that had higher concentration.CONCLUSION: In diabetes condition, complications of vascular calcification are caused by the mechanism of increased AGE formation represented by esRAGE. In diabetes control it is very important to keep the blood vessels from complications caused by vascular calcification.KEYWORDS: type 2 diabetes mellitus, vascular calcification, esRAGE, MGP, HbA1
Adiposopathy and Obesity Paradox
Get PDFBACKGROUND: Obesity has reached global epidemic proportions in both adults and children and is associated with numerous comorbidities, including hypertension, type 2 diabetes mellitus (T2DM), dyslipidemia and major cardiovascular diseases (CVD).CONTENT: Adiposity may cause adipocyte and adipose tissue anatomic and functional abnormalities, termed adiposopathy (adipose-opathy) or "sick fat," that result in endocrine and immune derangements. Adiposopathy may directly contribute to CVD through pericardiac and perivascular effects on the myocardium and blood vessels. Adiposopathy may also indirectly contribute to CVD through promoting or worsening major CVD risk factors such as T2DM, high blood pressure, and dyslipidemia. Despite this adverse association, numerous studies have documented an obesity paradox in which overweight and obese people with established CVD, including hypertension, heart failure, coronary heart disease, and peripheral arterial disease, have a better prognosis compared with nonoverweight/nonobese patients. These paradoxical findings are made less paradoxical when the pathogenic potential of excessive body fat is assessed based on adipose tissue dysfunction rather than simply on increased fat mass alone.SUMMARY: Adiposopathy is defined as pathological adipose tissue function that may be promoted and exacerbated by fat accumulation (adiposity) and sedentary lifestyle in genetically susceptible patients. Adiposopathy is a root cause of some of the most common metabolic diseases observed in clinical practice, including T2DM, hypertension and dyslipidemia.KEYWORDS: adiposopathy, adiposity, obesity paradox, adipocyte dysfunction, adipose hypertrophy, adipose hyperplasi
Habitual Coffee Consumption Does Not Correlate with Blood Pressure, Inflammation and Endothelial Dysfunction but Partially Correlates with Oxidative Stress
Get PDFBACKGROUND: Coffee is the most widely consumed beverage in the world and has been known to have effects on cardiovascular system. Many researchers have examined the effects of coffee consumption on blood pressure (BP) and risk of cardiovascular disease (CVD), but their results were inconsistent and still remain a subject of controversy. Oxidative stress, inflammation, and endothelial dysfunction have been known as risk factors of hypertension and CVD. Those factors are also known to be affected by coffee consumption. The aim of this study was to examine the relationship between the effects of habitual coffee consumption on BP and to examine the role of oxidative stress (F2 isoprostane), inflammation (high sensitive C-reactive protein (hsCRP)) and endothelial dysfunction (asymmetric dimethylarginine (ADMA)).METHODS: This was a cross-sectional study in which 47 healthy, non-smoking men aged 30-60 years with varying coffee-drinking habits were enrolled. BP and blood/urine analysis of biomarkers were measured in the morning before activity. Coffee consumption was assessed using a questionnaire. The differences among variables were analyzed using ANOVA and the correlations between variables were analyzed using Kendall’s Tau correlation analysis.RESULTS: Habitual coffee consumption did not correlate with systolic/diastolic BP (r=-0.02; p=0.856 and r=0.15; p=0.230, respectively). Concentrations of ADMA and hsCRP were also not correlated with coffee consumption (r=0.03; p=0.764 and r=0.04; p=0.701, respectively). Coffee consumption only showed significant correlation with F2 isoprostane (r=0.34; p=0.004).CONCLUSION: BP was not affected by coffee consumption although coffee consumption has a significant correlation with F2 isoprostane. These findings suggest that correlation between coffee consumption and BP might be explained by other factors that were not included in this study.KEYWORDS: coffee, caffeine, cardiovascular disease, blood pressure, oxidative stress, inflammation, endothelial dysfunctio
Association of Cross Linked C-Telopeptide II Collagen and Hyaluronic Acid with Knee Osteoarthritis Severity
Get PDFBACKGROUND: This study was carried out to investigate the association of Cross Linked C-Telopeptide Type I & II Collagen (CTX-I and II) and hyaluronic acid (HA) with knee osteoarthritis (OA) severity.METHODS: Sixty menopause women with primary knee OA were enrolled in this study during their visits to the Outpatient Department. Patients with knee pain during weight bearing, active or passive range of motion, or tenderness with Kellgren-Lawrence (KL) grade of more than I were included. Patients with injury, inflammatory and metabolic diseases were excluded. Patients were put in a 10-hour fasting prior to withdrawal of morning blood samples for examinations of HA, CTX-I, interleukin 1 beta (IL-1β), and high sensitivity C reactive protein (hs-CRP) level. Second void morning urine specimens were taken for CTXII assessment. HA, CTX-I and II levels were measured by enzyme-linked immunosorbent assay.RESULTS: Sixty menopausal female patients were included in this study, 35 with KL grade II, 17 grade III, and 8 grade IV. Means of CTX-II were significantly different between subjects KL grade IV and III (p=0.021). Correlation of KL grade was significant with CTX-II (p=0.001, r=0.412) and HA (p=0.0411, r=0.269). KL grades were not significantly associated with CTX-I (p=0.8364, r=-0.0272); IL-1β (p=0.5773, r=0.0853) and hs-CRP (p=0.2625, r=0.1470).CONCLUSION: CTX-II and HA were associated with severity of knee OA, suggesting that CTX-II and HA can be used as marker for knee OA severity.KEYWORDS: CTX-II, hyaluronic acid, otestoarthritis, kne
MicroRNAs in Cardiometabolic Diseases
Get PDFBACKGROUND: MicroRNAs (miRNAs) are ~22-nucleotide noncoding RNAs with critical functions in multiple physiological and pathological processes. An explosion of reports on the discovery and characterization of different miRNA species and their involvement in almost every aspect of cardiac biology and diseases has established an exciting new dimension in gene regulation networks for cardiac development and pathogenesis.CONTENT: Alterations in the metabolic control of lipid and glucose homeostasis predispose an individual to develop cardiometabolic diseases, such as type 2 diabetes mellitus and atherosclerosis. Work over the last years has suggested that miRNAs play an important role in regulating these physiological processes. Besides a cell-specific transcription factor profile, cell-specific miRNA-regulated gene expression is integral to cell fate and activation decisions. Thus, the cell types involved in atherosclerosis, vascular disease, and its myocardial sequelae may be differentially regulated by distinct miRNAs, thereby controlling highly complex processes, for example, smooth muscle cell phenotype and inflammatory responses of endothelial cells or macrophages. The recent advancements in using miRNAs as circulating biomarkers or therapeutic modalities, will hopefully be able to provide a strong basis for future research to further expand our insights into miRNA function in cardiovascular biology.SUMMARY: MiRNAs are small, noncoding RNAs that function as post-transcriptional regulators of gene expression. They are potent modulators of diverse biological processes and pathologies. Recent findings demonstrated the importance of miRNAs in the vasculature and the orchestration of lipid metabolism and glucose homeostasis. MiRNA networks represent an additional layer of regulation for gene expression that absorbs perturbations and ensures the robustness of biological systems. A detailed understanding of the molecular and cellular mechanisms of miRNA-mediated effects on metabolism and vascular pathophysiology could pave the way for the development of novel diagnostic markers and therapeutic approaches.KEYWORDS: microRNA, lipid metabolism, glucose homeostasis, vascular endothelium, vascular smooth muscle, atherosclerosi
Molecular Mechanisms of Cardiovascular Aging
Get PDFBACKGROUND: The average lifespan of humans is increasing, and with it the percentage of people entering the 65 and older age group is growing rapidly and will continue to do so in the next 20 years. Within this age group, cardiovascular disease will remain the leading cause of death, and the cost associated with treatment will continue to increase. Aging is an inevitable part of life and unfortunately poses the largest risk factor for cardiovascular disease.CONTENT: We provide an overview of some of the molecular mechanisms involved in regulating lifespan and health, including mitochondria, telomeres, stem cells, sirtuins, Adenosine Monophosphate-activated Protein Kinase, Mammalian Target of Rapamycin and Insulin-like Growth Factor 1. We also provide future perspectives of lifespan and health, which are intimately linked fields.SUMMARY: Aging remains the biggest non-modifiable risk factor for cardiovascular disease. The biological, structural and mechanical changes in senescent cardiovascular system are thought to contribute in increasing incidence of cardiovascular disease in aging. Understanding the mechanisms contributing to such changes is therefore crucial for both prevention and development of treatment for cardiovascular diseases.KEYWORDS: cardiovascular aging, mitochondria, telomeres, sirtuin, stem cell
Cost-Effectiveness Analysis of Pharmacotherapy for Hematemesis-Melena Treatment in Hospitalized Patients with Hepatic Cirrhosis
Get PDFBACKGROUND: Acute variceal haemorrhage is a complication of cirrhosis that can be life threatening. It is a pharmacist’s duty to ensure therapeutic and pharmaceutical care which is not only safe and effective for the patient but also is cost-effective in order to attain improvement of the patient’s quality of life. Therefore, pharmacoeconomic evaluation especially cost-effectiveness analysis (CEA), which compares costs and consequences of drug therapy, is needed. This study was aimed to evaluate the therapeutic cost-effectiveness of hematemesis-melena treatment in hepatic cirrhotic patients.METHODS: A total of 42 patients receiving vitamin K and vitamin K-transamin were studied retrospectively from patients’ medical records in 2 years and analyzed with cost-effectiveness grid and average cost-effectiveness ratio (ACER) based on Child-Turcotte-Pugh (CTP) Score.RESULTS: Cost-effectiveness grid was dominant for vitamin K in patients with CTP Score A. ACER analysis showed a lower score for vitamin K in all patients included CTP Score classification. There was no significant difference in duration of cessation of bleeding treatment in patients with vitamin K compared with vitamin K-transamin in patients with CTP Score A and B, while significant difference was found in patients with CTP Score C.CONCLUSION: Vitamin K appeared to be more cost effective as compared with vitamin K-transamin in all patients. The use of vitamin K had greater benefit than the combination with transamin in all patients and CTP Score classification, and thus should be considered as a primary therapy. Therefore, transamin addition as an alternative therapy for hepatic cirrhosis patients with hematemesis-melena should be considered.KEYWORDS: CEA, cost-effectiveness analysis, child-turcotte-pugh score, hepatic cirrhosis, hematemesismelena, vitamin K, transami
Current Issues and Challenges in the Use of Aerosolized Surfactant for Respiratory Distress Syndrome in the Newborns
Get PDFBACKGROUND: Surfactant replacement therapy is a recognized treatment for respiratory distress syndrome (RDS) in the newborns. Over the past 30 years, human and animal trials have been performed regarding administration of aerosolized surfactant to the injured lung, however the result has been unsatisfactory when compared with instilled surfactant delivery via endotracheal tube (ETT). This review aims to investigate the current issues, challenges and future recommendation of aerosolized surfactant therapy.CONTENT: Five randomized clinical trials in humans and 13 animal trials met the inclusion criteria and were reviewed. Most animal trials agree that this method of treatment is feasible. However, human trials presented conflicting results, and generally showed it to be ineffective. When compared with surfactant delivery via ETT, aerosolized surfactant is less effective in improving respiratory function.SUMMARY: The current data from human trials does not support the implementation of aerosolized surfactant therapy to treat newborns with RDS. Further research is necessary to improve nebulization, delivery, distribution and deposition in the lung, to investigate aerosolized surfactant delivery via ETT and to determine the appropriate dose.KEYWORDS: surfactant, aerosol, prematurity, respiratory distress syndrom