The Indonesian Biomedical Journal (Prodia Education and Research Institute)
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High VEGF Level is Produced by Human Umbilical Cord- Mesenchymal Stem Cells (hUC-MSCs) in Amino Acid-Rich Medium and under Hypoxia Condition
Get PDFBACKGROUND: Secretome production by stem cells depends on their culture conditions such as oxygen concentration and the composition of the culture media. In this study, we investigated the secretion of neurotrophic growth factors of human umbilical cord mesenchymal stem cells (hUC-MSCs) in amino acid-rich culture medium and under hypoxic condition.METHODS: hUC-MSCs were cultured in normoxic and various hypoxic (1%, 5%, 10%) conditions in an amino acid-rich culture medium. The end-point parameters (cell proliferation and survival, cell morphology and growth factor secretion) were measured at 3 time-points (48 hours, 72 hours and 96 hours). ELISA-based methods were used for neurotrophic factors detection, including neurotrophic growth factor (NGF), vascular endothelial factor (VEGF), and brain-derived neurotrophic factor (BDNF).RESULTS: NGF secretion was not detectable at any time points both in normoxia and hypoxia. BDNF secretion under normoxia was induced at 48 h time point and reached the highest level at an average of 181.9±13.01 pg/mL at 96 hours, whereas hypoxia exposure to hUC-MSCs only induced the BDNF secretion at low level. VEGF secretion was barely detectable in normoxic condition. However, VEGF secretion reached the highest level at an average of 7707.55±2110.85 pg/mL in 5% hypoxia at 96 hours.CONCLUSION: Combination of amino acid-rich culture medium and hypoxia condition dramatically induced high VEGF secretion by hUC-MSCs, especially at 5% hypoxia, induced mild BDNF secretion and had no effect toward NGF secretion.KEYWORDS: human umbilical cord mesenchymal stem cells, neurotrophic growth factor, amino acid-rich, hypoxi
High Expression of FcγII (CD32) Receptor on Monocytes in Dengue Infected Patients
Get PDFBACKGROUND: 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.KEYWORDS: dengue infection, FcγII (CD32) receptor monocyte, flow cytometr
Advanced in Molecular Mechanisms of Atherosclerosis: From Lipids to Inflammation
Get PDFBACKGROUND: Atherosclerosis is a leading cause of vascular disease worldwide. During the past several decades, landmark discoveries in the field of vascular biology have evolved our understanding of the biology of blood vessels and the pathobiology of local and systemic vascular disease states and have led to novel disease-modifying therapies for patients. This review is made to understand the molecular mechanism of atherosclerosis for these future therapies.CONTENT: Advances in molecular biology and -omics technologies have facilitated in vitro and in vivo studies which revealed that blood vessels regulate their own redox milieu, metabolism, mechanical environment, and phenotype, in part, through complex interactions between cellular components of the blood vessel wall and circulating factors. Dysregulation of these carefully orchestrated homeostatic interactions has also been implicated as the mechanism by which risk factors for cardiopulmonary vascular disease lead to vascular dysfunction, structural remodeling and, ultimately, adverse clinical events.SUMMARY: Atherosclerosis is a heterogeneous disease, despite a common initiating event of apoB-lipoproteins. Despite of acute thrombotic complications, an adequate resolution response is mounted, where efferocytosis prevents plaque necrosis and a reparative scarring response (the fibrous cap) prevents plaque disruption. However, a small percentage of developing atherosclerotic lesions cannot maintain an adequate resolution response, which leading to the formation of clinically dangerous plaques that can trigger acute lumenal thrombosis and tissue ischemia and infarction.KEYWORDS: atherosclerosis, oxidative stress, inflammation, efferocytosis, foam cells, thrombosi
Effect of Zinc and Iron Supplementation on Appetite, Nutritional Status and Intelligence Quotient in Young Children
Get PDFBACKGROUND: Lack of appetite in young children leads to growing incidences of physical and mental growth disorders. Supplementation of certain micronutrients can increase appetite and improve nutritional status. This study aims to analyze the effects of zinc and iron supplementation on appetite, nutritional status and intelligence quotient (IQ) in young children.METHODS: An experimental study withrandomized control group pre/post-test design was conducted in Semarang, Indonesia. A total of 68 children were divided into four groups. The first group was the control group, which was given a placebo; the second group was given a zinc supplement at 10 mg/day; the third group was given an iron supplement at 7.5 mg/day; andthe fourth group was given zinc and iron for three months. Appetite was assessed based on eating frequency and energy intake. Nutritional status was assessed by weight per age (W/A) and height per age (H/A) z score. IQ score was assessed based on Wechsler Preschool and Primary Scale of Intelligence (WPPSI).RESULTS: Before intervention, low zinc intake was observed in 27.7% of the subjects and low iron intake was observed in 58.5% of them. After intervention, appetite in the second and fourth groups increased. W/A z score increased in the second and third groups. IQ score increased in the third group. No significant effect on H/A z score was observed in all groups.CONCLUSION: Supplementation of zinc and iron for three months had a positive effect on appetite, body weight and IQ score but no significant effect on body height.KEYWORDS: appetite, zinc, iron, growt
Caffeic Acid Inhibits RANKL and TNF-α-induced Phosphorylation of p38 Mitogen-activated Protein Kinase in RAW-D Cells
Get PDFBACKGROUND: Caffeic acid inhibits osteoclastogenesis by downregulating expression of Cathepsin K and Nuclear Factor of Activated T cells (NFATc)1, as well as inhibiting activity of Nuclear Factor kB (NFkB). Meanwhile TNF Receptor-associated Factor (TRAF)6 was not influenced by caffeic acid. In order to investigate further caffeic acid's mechanism in inhibiting osteoclastogenesis, regulation of caffeic acid on p38 Mitogen-activated Protein Kinase (MAPK) was investigated.METHODS: RAW-D cells were pretreated with/without caffeic acid and treated with/without 20 ng/mL RANKL and 1 ng/mL TNFα for 0.2, 1, 6, and 12 hour. Tartrate Resistant Acid Phosphatase (TRAP) staining was performed. Then, western blot analysis was performed to detect p38 MAPK and phosphorylated-p38 MAPK. Resulted protein bands were quantified and statistically analyzed.RESULTS: Under induction of 20 ng/mL RANKL and 1 ng/mL TNF-α, RAW-D cells were successfully differentiated into TRAP+ osteoclast-like polynuclear cells. Under treatment of 20 ng/mL of RANKL and 1 ng/mL of TNF-a for 0.2 or 1 hour, significant (p=0,000, T test) increment of phosphorylated p38 MAPK was observed as compared with control. Pretreatment of 10 μg/mL caffeic acid significantly (p=0.000, T test) suppressed the 20 ng/mL of RANKL and 1 ng/mL of TNF-a-induced phosphorylation of p38 MAPK.CONCLUSION: RANKL and TNF-a are potent osteoclastogenesis inductors in RAW-D cells, meanwhile caffeic acid could inhibit the RANKL and TNFa-induced osteoclastogenesis through p38 MAPK.KEYWORDS: caffeic acid, osteoclastogenesis, RANKL, TNF-a, p38, MAPK, RAW-D cell
The Effect of Collagen Activation on Platelet Rich Plasma for Proliferation of Periodontal Ligament Fibroblasts
Get PDFBACKGROUND: Regenerative procedure in periodontal surgery aims to improve the structure and function of the periodontium to be strong enough to support the teeth. Growth factor is essential in the process of tissue regeneration; it can be generated from the activation of Platelet-Rich Plasma (PRP). In this study, collagen was used as PRP activator. PRP release growth factor from the granules when activated. The aim of this study was to evaluate the effect of collagen on PRP activation and the collagen-activated PRP storage to fibroblast proliferation of periodontal ligament (PDL).METHODS: Fibroblasts of PDL were obtained from extracted premolar. PRP were obtained from 100 mL of blood donors using double centrifugation methods. PRP were activated by collagen and subsequently incubated for 24, 48, 72 and 168 hours; after which the lysate was taken. Fibroblasts were divided into 7 groups, which consisted of one unstimulated group as negative control, one group stimulated by PRP lysate, and five groups stimulated with PRP-collagen lysates that had been incubated for 24, 48, 72 and 168 hours. MTT assay was then performed after 1 and 3 days.RESULTS: The proliferation rate of fibroblasts group stimulated by PRP-collagen lysate was higher than the group stimulated by PRP. The storage of collagen-activated PRP for 7 days at 4°C could increase the proliferation rate.CONCLUSION: The activation of collagen on PRP and the stored collaged-activated PRP could increase the fibroblasts proliferation rate of PDL.KEYWORDS: collagen, platelet-rich plasma, fibroblasts proliferatio
Effect of Puguntano Extract (Curanga Fel-Terrae Merr.) on hs-CRP Level in Newly Diagnosed Type 2 Diabetes Mellitus Patient
Get PDFBACKGROUND: Inflammation is one of the factors that associated with insulin resistance which causes type 2 diabetes mellitus (T2DM). Inflammation can be measured by using high sensitivity C-reactive protein (hs-CRP), a quantitative measurement of CRP concentration. Puguntano is a plant that traditionally used for antidiabetic treatment found in North Sumatera. The objective of this study is to know the effect of Puguntano extract on hs-CRP level in newly diagnosed T2DM patient.METHODS: The study was a randomized controlled clinical trial on 24 newly diagnosed T2DM patients. Subjects were divided into 2 groups. The first group was given Puguntano extract with starting doses at 100 mg daily, then the dose was increased in titration up to a maximum dose of 2x100 mg daily depending on blood sugar level monitoring. Meanwhile, the other group (which serves as a control group) was given metformin with starting doses at 500 mg daily, then the dose was increased in titration up to a maximum dose of 3x500 mg per day depending on blood sugar level monitoring. The hs-CRP and HbA1c are assessed at the baseline and after 12 weeks of treatment.RESULTS: The hs-CRP level was decreased in Puguntano group by 1.41 (-7.54-0.52) (p=0.06) and decrease by 0.58 (-7.86-33.41) (p=0.695) in metformin group. There was no difference in hs-CRP level between the two groups (p=0.630).CONCLUSION: Puguntano extract showed the effect of lowering HbA1C and hs-CRP level in T2DM patients, although it was not statistically significant.KEYWORDS: type 2 diabetes mellitus, hs-CRP, puguntan
The dtxR Gene: A New Alternative Marker to Identify Corynebacterium ulcerans and Corynebacterium pseudotuberculosis by PCR Assay
Get PDFBACKGROUND: There are found some studies which reported the successfull of polymerase chain reaction (PCR) assay to identify Corynebacterium ulcerans and Corynebacterium pseudotuberculosis. This study aimed to describe the dtxR gene profile as a new marker for C. ulceran and C. pseudotuberculosis for the PCR assay.METHODS: Ten C. ulcerans and 35 C. pseudotuberculosis DNA sequences data registered in GeneBank was analyzed by bioinformatic tools. PCR primer was designed based on the concerved region and the gene similarity data. On the other hands, reference strains (C. ulcerans NCTC 12077 and Corynebacterium diphtheriae NCTC 3984) and dtxR gene of C. pseudotuberculosis (synthetic gene) were used in the PCR assay optimization for C. ulcerans and C. pseudotuberculosis identification.RESULTS: The study showed that dtxR genes of both C. ulcerans and C. pseudotuberculosis were more conserve than pld gene, moreover dtxR gene was more specific compared to 16S rRNA gene. PCR assay with dtxR gene as a target could identify C. ulcerans and C. pseudotuberculosis accurately without mispriming, misamplification and misidentification.CONCLUSION: dtxR gene could be used as marker to identify C. ulcerans and C. pseudotuberculosis by PCR assay.KEYWORDS: C. pseudotuberculosis, C. ulcerans, dtxR gene, PC
Immunomodulatory Effect of Momordica charantia L. Fruit Ethanol Extract on Phagocytic Activity and Capacity of Mice Peritoneal Macrophages
Get PDFBACKGROUND: The purpose of this research is to understand the secondary metabolites of Momordica charantia L. extract, as well as to disclose the potential of M. charantia extract in the phagocytic activity and capacity of peritoneummacrophages.METHODS: Examination of immunomodulatory effect was done by giving M. charantia ethanol extract on 5 treatment groups, given intra-peritoneally to mice daily. Echinacea extract as positive control and double distilled water as negative control were also given. On the 8th day, mice were infected with Staphylococcus epidermidis. After 30 minutes, peritoneum fluid was obtained to observe the activity and capacity of macrophage cells.RESULTS: The results showed significant phagocytic activity (p0.05). The highest phagocytic activity was the group treated with 1,200 ppm (62%), significantly higher than other groups.CONCLUSION: The secondary metabolite content of M. charantia is alkaloids, flavonoids, tannins, saponins, and triterpenoids. The 1,200 ppm M. charantia ethanol extract is potential in inducing phagocytic activity and capacity. These results indicate that the M. charantia can be suggested as a natural immunomodulator.KEYWORDS: pare fruit, Momordica charantia L., phagocytosis, macrophage, immunomodulato
New Insight in The Molecular Mechanisms of Neurodegenerative Disease
Get PDFBACKGROUND: Redox and proteotoxic stress contributes to age-dependent accumulation of dysfunctional mitochondria and protein aggregates, and is associated with neurodegeneration. The free radical theory of aging inspired many studies using reactive species scavengers such as alpha-tocopherol, ascorbate and coenzyme-Q to suppress the initiation of oxidative stress. However, clinical trials have had limited success in the treatment of neurodegenerative diseases (NDDs).CONTENT: The misfolding and aggregation of specific proteins is a seminal occurrence in a remarkable variety of NDDs. In Alzheimer’s disease, the two principal aggregating proteins are β-amyloid (Aβ) and tau. The abnormal assemblies formed by conformational variants of these proteins range in size from small oligomers to the characteristic lesions that are visible by optica lmicroscopy, such as senile plaques and neurofibrillary tangles. Pathologic similarities with prion disease suggest that the formation and spread of these proteinaceous lesions might involve a common molecular mechanism, corruptive protein templating. The accumulation of redox modified proteins or organelles cannot be reversed by oxidant intercepting antioxidants and must then be removed by alternative mechanisms. Autophagy serves this essential function in removing damaged or dysfunctional proteins and organelles thus preserving neuronal function and survival.SUMMARY: Senescent cells and their senescence-associated secretory phenotypes (SASPs) may constitute a novel, understudied, and potentially important contributor to neuro-inflammation and subsequent neurodegeneration. Characterization of cellular senescence in the brain could uncover novel therapeutic targets for the prevention and treatment of chronic age-related NDDs.KEYWORDS: brain, aging, neurodegeneration, DNA damage, senescence, neuro-inflammation, mitochondria, lysosome, proteostasis, prion, amyloidosi