The Indonesian Biomedical Journal (Prodia Education and Research Institute)
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Comparison of Tumor Necrosis Factor-α Level in Coronary Artery Disease and Coronary Slow Flow of Thrombolysis in Myocardial Infarction
Get PDFBACKGROUND: 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 (r0.05).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.KEYWORDS: TNF-alpha, coronary artery disease, coronary slow flow
Conditioned Media of Human Umbilical Cord Blood Mesenchymal Stem Cell Inhibits Ultraviolet B-induced Apoptosis in Fibroblasts
Get PDFBACKGROUND: Ultraviolet (UV)B irradiated-skin cells may respond either by activating protective or apoptotic mechanisms. Several treatments has been reported to prevent apoptosis of the skin cell. To date, despite of the cell, secretome of mesenchymal stem cell (MSC) has been progressively explored for various treatment. Current research was conducted to find out the possible effect of MSC's secretome to protect normal cells from the UVB irradiation.METHODS: For preparation of the conditioned media human umbilical cord blood mesenchymal stem cell (CM-hUCB-MSC), hUCB was collected and separated to collect mononuclear cell (MNC). MNC was cultured in MSC medium until passage 4, then checked for MSC biomarkers. Conditioned media was then produced from the cultured MSC. For induction of apoptosis, NIH3T3 cells were pretreated with/without CM-hUCB-MSC, prior to the UVB irradiation for 5-60 minutes. After 24 hours, apoptosis evaluation was conducted with Sub-G1 assay with hypotonic fluorochrome solution using flow cytometer.RESULTS: The average of apoptotic cells irradiated with UVB for 30 minutes was 94.2%. The 30-minutes-UVB irradiation significantly induced apoptosis in NIH3T3 cells (p=0.000). The averages of apoptotic cells by pretreatment of 0, 5, 10 and 20% CM-hUCB-MSC prior to UVB irradiation were 94.2, 46.8, 31.8 and 31.5 %, respectively. The pretreatment of 5, 10 and 20% CM-hUCB-MSC prior to UVB irradiation could significantly decrease the percentage of apoptosis caused by UVB (p=0.001).CONCLUSION: Taken together, CM-hUCB-MSC inhibited UVB-induced apoptosis in NIH3T3 cells significantly, suggesting that CM-hUCB-MSC might be a potential anti-UVB-induced apoptosis factor. Hence, further research should be explored to disclose its specific intracellular mechanism.KEYWORDS: UVB, stem cell, conditioned media, fibroblast, apoptosis, aging, secretome, NIH3T3 cel
sodA and gap Genes as Markers for The Identification of Staphylococcus capitis
Get PDFBACKGROUND: 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.KEYWORDS: Staphylococcus, S. capitis, sodA, gap, PC
Clinicopathological Characteristic and Prognostic Significance of Indonesian Triple Negative Breast Cancer
Get PDFBACKGROUND: 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.KEYWORDS: NBC, clinicopathological characteristics, prognostic significance
Possible Renal Repairing Mechanisms of Mesenchymal Stem Cells in Cyclosporine-Mediated Nephrotoxicity: Endothelial Viability and Hemodynamics
Get PDFBACKGROUND: Stem cell-based therapy may represent the first realistic option for tissue repairing and regeneration. Mesenchymal stem cells (MSCs) are proved to be involved in the regeneration of many tissues which are subjected to different types of injury. Cyclosporine (CsA) in spite of its use as immune suppressive drug during kidney transplantation, it was considered as important model of nephropathy specially, during long-term administration.METHODS: Isolation and preparation of MSCs using Dulbecco's modified Eagle's medium (DMEM), in vitro differentiation through adipogenesis chondrogenesis and osteogenesis was confirmed by using Alizarin Red S stain, Oil Red O stain and Alcian Blue stain, respectively, characterization using flow cytometry technique to detect cluster of differentiation (CD)34, CD44 and CD105 surface markers and homing of MSCs using polymerase chain reaction (PCR) Sry gene assay, were executed. Serum levels of vascular endothelial growth factor (VEGF), endothelin-1 (ET-1) and endothelial nitric oxide synthase (eNOS) were quantified using enzyme-linked immunosorbent assay (ELISA) kits based on the principle of double-antibody sandwich technique. The structural organization of kidney tissue was examined using histological procedures.RESULTS: Single intravenous dose of MSCs is capable to boost kidney repairment process as indicated by the significant decrease in serum ET-1 level paralleled by significant increase in VEGF and eNOS. Moreover, histological findings revealed the improvement of focal hemorrhage in between the degenerated tubules, congestion in the cortical blood vessels, vacuolization in the endothelial cells lining the glomerular tufts and focal perivascular inflammatory cells aggregation.CONCLUSION: This study demonstrated the favorable influence of MSCs in repressing cyclosporine-induced nephropathy in rats. This could be achieved through angiogenic and anti-arteriolopathic mechanisms.KEYWORDS: angiogenesis, cyclosporine, endothelin-1, MSCs, nephropath
Combination of Aerobic Exercise and Continuous Environmental Enrichment Improves Adult Male Rats’ Spatial Memory: Study on Hippocampal Insulin Like Growth Factor 1 (IGF-1) and Fibroblast Growth Factor 2 (FGF-2) Expression
Get PDFBACKGROUND: Memory declines with the progression of age through the neurodegeneration process. Aerobic exercise and environmental enrichment can delay neurodegeneration by improving neuroplasticity via expression of insulin like growth factor 1 (IGF-1), fibroblast growth factor 2 (FGF-2) and other proteins. Combination treatment of aerobic exercise and continuous environmental enrichment and their effect on the expression of IGF-1 and FGF-2 which were expected to improve memory function has not been studied previously. Thus, this study aimed to observe it.METHODS: This is an experimental research using 24 male Wistar rats (Rattus norvegicus, 300-400 g, age 7-8 months) divided randomly into 4 groups: control (C), aerobic exercise (A), continuous (EE), and combination of aerobic exercise and continuous environmental enrichment (A-EE). At the end of an 8-week treatment, rats were sacrificed, and an enzyme-linked immunosorbent assay (ELISA) examination was performed to assess hippocampal IGF-1 and FGF-2 levels.RESULTS: In the 8th week, A-EE group showed the best improvement in rats’ spatial memory (47.84±10.6 %) followed by EE group (45.03±4.1 %), A group (38.61±3.8 %), and C group (22.76±7.12 %). However, A-EE group’s hippocampal IGF-1 (16.21±7.56 ng/mg protein) and FGF-2 (1.29±0.57 ng/mg protein) expression were not higher than other groups.CONCLUSION: Improvement in memory function in the combination group is a result of induction of various growth factors’ expression in the hippocampus, including IGF-1 and FGF-2, but the primary pathway of memory function improvement may be from other growth factors.KEYWORDS: spatial memory, aerobic exercise, environmental enrichment, hippocampus, IGF-1, FGF-
High Myostatin Serum Related with High Prevalence of Sarcopenia Among Elderly Population in Pedawa Village, Bali, Indonesia
Get PDFBACKGROUND: Sarcopenia is defined as a decrease in muscle mass accompanied by a decrease in muscle strength and performance. Sarcopenia arises from the disruption of the complex balance between anabolic and catabolic factors. Myostatin strongly influences muscle growth inhibition. Deletion and function loss of myostatin causes hyperplasia and skeletal muscle hypertrophy.METHODS: This study was an analytical cross-sectional study. Seventy respondents aged ≥60 years in Pedawa Village, Bali, Indonesia were selected by using the stratified random sampling technique. Sarcopenia status was assessed according to Asian Working Group for Sarcopenia (AWGS) criteria, including muscle mass, grip strength, and walking speed. While the myostatin serum levels was measured by enzyme-linked immunosorbent assay (ELISA).RESULTS: The incidence of sarcopenia in the elderly was 45 people (64.3%). Based on the analysis, there was a significant difference between myostatin levels in sarcopenia subjects (47.59 ng/mL) and non-sarcopenia subjects (39.7 ng/mL). Based on the statistical calculations, it was determined that the cut-off range of myostatin levels was 48.91 ng/mL. The prevalence ratio of sarcopenia incidence based on the myostatin levels in the elderly was 3.84, while based on the combination of age risk and myostatin levels was 9.75.CONCLUSION: Based of the data, there are significant differences of myostatin level between elderly people with and without sarcopenia. The prevalence of high myostatin levels in elderly is almost 4 times higher than low myostatin levels in the elderly.KEYWORDS: myostatin, sarcopenia, elderl
The Effect of Tomato Juice in Increasing Ki-67 Expression and Epithelial Thickness on The Vaginal Wall of Menopausal Rats
Get PDFBACKGROUND: 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.KEYWORDS: epithelial thickness, Ki-67, menopause, tomato juic
Mesenchymal Stem Cell Secretome: Cell-free Therapeutic Strategy in Regenerative Medicine
Get PDFBACKGROUND: 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.KEYWORDS: MSC, secretome, conditioned media, extracellular vesicle, exosom
Artificial Intelligent in Healthcare
Get PDFBACKGROUND: Giant transformations are going on currently in health care, and the greatest force behind this phenomenon is data.CONTENT: Big data has arrived into medicine field, lead to potential enhancement in accountability, quality, efficiency, and innovation. Most updated, artificial intelligence (AI) and machine-learning (ML) techniques rapidly developed, bring forth the big data analysis into more useful applications, from resource allocation to complex disease diagnosis. To realize this, a very large set of health-care data is needed for algorithms training and evaluation, including patients’ treatment data, patients respond to treatment, and personal patient information, such as genetic data, family history, health behavior, and vital signs.SUMMARY: Precision Health involving preventive, predictive, personalized and precise. The arrival of AI and ML will enhance and facilitates the improvement of this relationship through better accuracy, productivity, and workflow, thus develop a health system that will go beyond just curing disease, but further into wellness that preventing disease before it strikes, thus the patient–doctor bond is expected to be reformed and not be eroded.KEYWORDS: artificial intelligence, machine learning, deep learning, electronic health records, big dat