The Indonesian Biomedical Journal (Prodia Education and Research Institute)
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Screening of Extended-Spectrum β-Lactamases (ESBL)-producing Klebsiella pneumoniae with ChromID ESBL Media
Get PDFBACKGROUND: 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.KEYWORDS: K. pneumoniae, ChromID ESBL, DDST, ESBL, sensitivit
Analysis of ALDH1A1 and ALDH1A3 Gene mRNA Expressions in Adipose-Derived Stem Cells (ASCs) and Umbilical Cord Stem Cells (UCSCs)
Get PDFBACKGROUND: ALDH1A1 and ALDH1A3, the most renowned isozymes of aldehyde dehydrogenase (ALDH)1, are important in regulating the pluripotency of human mesenchymal stem cells (MSCs) and cancer stem cells (CSCs). The study aimed to analyze the mRNA expression of ALDH1A1 and ALDH1A3 genes in adipose stem cells (ASCs) and umbilical cord stem cells (UCSCs) along with their correlations to Oct-4 mRNA expression. Additionally, the interaction between these proteins was also investigated using in silico study to confirm the pluripotency of both MSCs compared to human breast ALDH+ CSCs.METHODS: This research focused on determining mRNA levels of ALDH1A1, ALDH1A3 and Oct-4 in ASCs and UCSCs using one-step qRT-PCR. The data were then normalized to those in human breast CSCs and 18S rRNA. Oct-4 gene expression was also analyzed to determine the pluripotency of ASCs and UCSCs. The protein-protein interactions were in silico analyzed using String 9.1 software.RESULTS: Relatively, ALDH1A3 was expressed at similar level in ASCs and UCSCs, while ALDH1A1 expression level was significantly higher in ASCs compared to UCSCs. In contrast to ALDH1A3, the expressions of ALDH1A1 in both MSCs were significantly lower than breast CSCs similar to Oct-4 expressions, as also revealed by the in silico data showing the interaction between these proteins. This suggests the role of ALDH1A1 on pluripotency.CONCLUSION: ALDH1A1 and ALDH1A3 were distinctly expressed in UCSCs and ASCs, which might be associated with unique properties of ASCs and UCSCs. This study may contribute to further research in terms of implication of ALDH1A1 and ALDH1A3 expressions towards the properties of MSCs and its application in stem cell therapy.KEYWORDS: ALDH1A1, ALDH1A3, UCSC, ASC, breast ALDH+ CSC, Oct-
Positive Correlation between Very Small Embryonic Stem Cell, Hematopoietic Stem Cell, and Endothelial Progenitor Cell in Umbilical Cord Blood Unit
Get PDFBACKGROUND: 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.KEYWORDS: stem cell, umbilical cord blood, endothelial progenitor, flowcytometr
The Effect of Estradiol on Renal Function in Reversible and Irreversible Unilateral Ureteral Obstruction in Rats
Get PDFBACKGROUND: The ureteral obstruction (UO) is the most common clinical disorders. This study was designed to investigate the renoprotective effect of estradiol in unilateral UO (UUO) and reversible UUO (RUUO).METHODS: Eighty-four ovareictomized Wistar rats were assigned into 14 groups. Group 1 was sham operated group. All the groups 2-14 were subject to UUO, however they received additional treatments. Group 2-4 received test substances (vehicle, 0.1 or 0.05 mg/kg of estradiol) and sacrificed after 3 days. Group 5-7 received test substances and after 3 days followed by RUUO (obstruction removal) and after one day post RUUO were sacrificed. Group 8-9 followed by RUUO after 3 days and simultaneously received estradiol and after one day post RUUO were sacrificed. Group 10-12 followed by RUUO after 3 days and simultaneously received test substances and after 3 days post RUUO were sacrificed. Group 13-14 received test substances followed by RUUO and received test substances again and after 3 days post RUUO were sacrificed.RESULTS: The estradiol reduced body weight significantly (p<0.05) in UUO, and the uterus weight (UW) increased significantly by estradiol (p<0.05). The increased serum levels of blood urea nitrogen (BUN) and creatinine (Cr), and kidney weight (KW) induced by UUO (p<0.05) were not attenuated by estradiol. Estradiol treatment did not alter the levels of BUN, Cr, KW, Cr clearance (CrCl), urine sodium excretion and urine flow when compared with control.CONCLUSION: Although the protective role of estradiol in cardiovascular system is well accepted, however our findings suggest that its protection during UUO or after RUUO in renal system cannot be certain.KEYWORDS: estradiol, ureteral obstruction, renal function, ra
UV-Visible Spectrophotometric as a Prospective Tool in Neonatal Sepsis
Get PDFBACKGROUND: This study was aimed to employ the UV-Visible (UV-Vis) spectrophotometry techniques to detect the changes in the blood of neonatal sepsis (NS) subject for a deeper understanding in the pathomechanism of NS.METHODS: The cross-sectional study was conducted from February to May 2017 in the Neonatology Division, Department of Pediatric, Ulin General Hospital/Faculty of Medicine, Lambung Mangkurat University, Banjarmasin, South Kalimantan, Indonesia. Blood specimens were taken from newborns, of which 15 each of newborns at risk of sepsis and without risk of sepsis. Data were analyzed by using Mann-Whitney U test.RESULTS: The result of this study suggested that there is a significant difference in the average of absorbance ratio parameter using UV-Vis spectrophotometric methods on the case group compared to the control group. Also, there is a significant difference between advanced oxidation protein products (AOPPs) and thiocyanate (SCN) level in newborn at risk of sepsis.CONCLUSION: The present study demonstrated there were significant differences between the average of absorbance ratio parameter for protein and oxy-hemoglobin region using UV-Vis spectrophotometric methods in healthy subjects and newborn at risk of sepsis.KEYWORDS: spectrophotometric, neonatal sepsis, oxidative stres
Anthropometry-based Body Fat Percentage Predicts High hs-CRP in Chronic Kidney Disease Patients
Get PDFBACKGROUND: Obesity is an important cardiovascular risk factor and associated with low grade inflammation in chronic kidney disease (CKD) patients. This study aims to assess the association between body fat with serum high sensitivity C-reactive protein (hs-CRP) level in CKD patients.METHODS: A cross-sectional study was performed in 71 CKD patients. Anthropometric measurements included body weight, height, body mass index (BMI), body fat percentage (BFP), skinfold thickness (SKF) of triceps and biceps were performed by trained physician. BFP was calculated using Kwok’s Formula and hs-CRP was measured by Particle enhanced Turbidimetry.RESULTS: The averaged BMI of our subjects was 25.8±4.4. There was no significant difference in BMI between pre-dialysis and hemodialysis CKD patients. Positive correlation was found between BFP and hs-CRP (r=0.266; p<0.05), while there was no significant correlation between BMI and hs-CRP.CONCLUSION: Body fat percentage was associated with hs-CRP. Hence, it will be more beneficial to assess nutritional status in CKD using BFP rather than BMI alone since it was demonstrated to correlate with hs-CRP in our studyKEYWORDS: CKD, obesity, inflammation, body fat, hs-CR
Effect of Electrical Stimulation in Lower Extremity as Physical Exercise in Type 2 Diabetes Mellitus Patients
Get PDFBACKGROUND: Type 2 diabetes mellitus (T2DM) is a long-term illness that affects the patient's quality of life and requires substantial health care costs. Lifestyle relatedto physical exercise is one of the causes of the increasing prevalence of this disease. However, not all T2DM patients can perform physical exercise because very weak physical conditions such as elderly, spinal cord injury or diabetic secondary complications can be contraindicated when physical exercise performed. The aim of this study is to observe the used of electrical stimulation in T2DM patients.METHODS: The provision of ES to lower extremity muscles was given for 30 minutes in the intervention group. Blood glucose samples were taken before and after the ES intervention. The data collected was statistically analyzed by using paired T-test and expressed in mean±standard error.RESULTS: The ES intervention decrease blood glucose level significantly from 193.03±5.740 mg/dL to 170.66±5.200 mg/dL (p≤0.001) in the last session.CONCLUSION: The effect of ES in lower extremity is shown to be significantly lowers the blood glucose level in T2DM patients.KEYWORDS: blood glucose, electrical stimulation, physical exercise, type 2 diabetes mellitu
Intervertebral Disc Degeneration and Low Back Pain: Molecular Mechanisms and Stem Cell Therapy
Get PDFBACKGROUND: Low back pain (LBP) mostly caused by disc degeneration, reflects to a tremendous of health care system and economy. More knowledge about these underlying pathologies will improve the opportunities that may represent critical therapeutic targets.CONTENT: Basic research is advancing the understanding of the pathogenesis and management of LBP at the molecular and genetic levels. Cytokines such as matrix metalloproteinases, phospholipase A2, nitric oxide, and tumor necrosis factor-α are thought to contribute to the development of LBP. Mesenchymal stem cells (MSCs) transplant to cartilage-like cells and secrete extracellular matrix and encourage nucleus pulposus (NP) cell activity inhibiting NP cell apoptosis, together with some chemical mediators such as cytokines and growth factors become a safe and effective new strategy for intervertebral disc degeneration (IDD) treatment and regeneration.SUMMARY: IDD occurs where there is a loss of homeostatic balance with a predominantly catabolic metabolic profile. A basic understanding of the molecular changes occurring in the degenerating disc is important for practicing clinicians to help them to inform patients to alter lifestyle choices, identify beneficial or harmful supplements, or offer new biologic, genetic, or stem cell therapies.KEYWORDS: low back pain, intervertebral disc, degeneration, nucleus pulposus, annulus fibrosus, extracellular matrix, genetic, stem cell
Nuclear Factor Erythroid 2-related Factor 2 (Nrf2) as A Therapeutical Target in Type-2 Diabetes Mellitus: A Review
Get PDFBACKGROUND: Increasing free radicals and oxidative stress due to chronic hyperglycemia in type-2 diabetes mellitus (DM) promotes the activity of endogenic antioxidative genes. Nuclear factor erythroid 2-related factor 2 (Nrf2) expression and activity are important to regulate the production of endogenic antioxidative enzymes.CONTENT: Normally, Nrf2 is bound by protein Kelchlike ECH-associated protein-1 (Keap1) in the cytosol. Stimulation from oxidative stress causes the release of Nrf2 from Keap1. When activated, Nrf2 enters the nucleus and activates the antioxidant response element (ARE). This will further increase the production of antioxidative enzymes, such as catalase, nitrite oxydase and heme oxygenase-1. The discovery of oxidative stress, as the cause of complications in DM, gives rise to the idea of developing a treatment which can increase the expression and activity of Nrf2, one of which is a flavonoid antioxidant.SUMMARY: Currently, nuclear factor erythroid 2-related factor 2 is an important target in the therapy of DM.KEYWORDS: Nrf2, type-2 diabetes mellitus, endogenic antioxidative enzymes, oxidative stress, antioxidant
Genome Editing with Crispr-Cas9 Systems: Basic Research and Clinical Applications
Get PDFBACKGROUND: Recently established genome editing technologies will open new avenues for biological research and development. Human genome editing is a powerful tool which offers great scientific and therapeutic potential.CONTENT: Genome editing using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPRassociated protein 9 (Cas9) technology is revolutionizing the gene function studies and possibly will give rise to an entirely new degree of therapeutics for a large range of diseases. Prompt advances in the CRISPR/Cas9 technology, as well as delivery modalities for gene therapy applications, are dismissing the barriers to the clinical translation of this technology. Many studies conducted showed promising results, but as current available technologies for evaluating off-target gene modification, several elements must be addressed to validate the safety of the CRISPR/Cas9 platform for clinical application, as the ethical implication as well.SUMMARY: The CRISPR/Cas9 system is a powerful genome editing technology with the potential to create a variety of novel therapeutics for a range of diseases, many of which are currently untreatable.KEYWORDS: genome editing, CRISPR-Cas, guideRNA, DSB, ZFNs, TALE