The Indonesian Biomedical Journal (Prodia Education and Research Institute)
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Lower GCS is Related to Poor Outcome among Acute Stroke Patients with COVID-19 in A Tertiary Referral Hospital in Indonesia
Get PDFBACKGROUND: The Coronavirus disease 2019 (COVID-19) pandemic has forced the health workforce to take mitigative measures such as physical distancing, screening, personal protective equipment donning, and confinement on patient care. We aimed to study the outcome of acute stroke patients with suspected, probable, or confirmed COVID-19 in a tertiary referral hospital in Indonesia during the first year of COVID-19 pandemic.METHODS: This was a retrospective study examining all medical records of adult patients suffering from acute stroke with suspected or confirmed COVID-19 who were admitted to R.D. Kandou Hospital, Manado, Indonesia, between March 2020 to March 2021. Clinical and laboratory parameters were compared between subjects with poor and good outcomes based on Glasgow Outcome Scale (GOS), divided into poor outcome (GOS 1-3) and good outcome (GOS 4-5).RESULTS: Fourty-six eligible subjects were enrolled in the study. Based on the GOS, 36 subjects (78.3%) were admitted to the hospital with poor prognosis. On admission, the median Glasgow Coma Scale (GCS) was 11, breathlessness was found in 54.3% of subjects, fever was found in only 15 subjects (32.6%), and the lowest oxygen saturation on admission 95%. We found that GCS significantly related to outcome after controlled for other factors using the logistic regression method (p=0.03; 95% CI=1.08-4.78).CONCLUSION: Lower GCS can be used to predict poor outcome in acute stroke patients with COVID-19.KEYWORDS: COVID-19, acute stroke, Glasgow Coma Scale, outcome, Indonesi
Soluble Transferrin Receptor and Soluble Transferrin Receptor/Log Ferritin Ratio are Correlated with Iron Status in Regular Hemodialysis Patients
Get PDFBACKGROUND: Monitoring of iron status in chronic kidney disease patients is important, however inflammation may hinder its monitoring. Soluble transferrin receptor (sTfR) is an alternative parameter to overcome this issue, whereas ferritin play a part in the inflammation process. Hence, the correlation between the sTfR ratio and the sTfR/log ferritin ratio with conventional iron status parameters in regular hemodialysis patients is necessary to be evaluated.METHODS: A cross-sectional was conducted in the current study. As many as 5 mL of blood (2 mL for sTfR and 3 mL for serum iron and ferritin levels) was collected. sTfR level was the blood-soluble transferrin receptor level measured by the enzyme-linked immunosorbent assay (ELISA). The amount of ferritin and serum iron was determined using the immunochemiluminescent process. To evaluate the correlation, the Pearson correlation test was used.RESULTS: A total of 80 subjects was included in this study. The mean of hemoglobin was 10.25±1.66 g/dL, serum iron was 58.19±26.56 g/dL, and the median ferritin was 520.4 (49.9-3606) ng/mL. The sTfR was significantly associated only with serum iron levels with a correlation coefficient of r=-0.242; p=0.031. The sTfR/log ferritin was significantly associated with serum iron l evels (InSI)(r=-0.255, p=0.022); and transferrin saturation (r=-0.295; p=0.008).CONCLUSION: sTfR/log ferritin has a negative and significant correlation with serum iron levels and transferrin saturation, while sTfR negatively correlated with serum iron levels. sTfR and sTfR/log ferritin may be considered as an alternative iron marker in inflammation setting such as CKD.KEYWORDS: sTfR/log ferritin, iron status, serum iron, ferritin, chronic kidney disease, hemodialysi
Obtaining Specific Hybridomas for Ki-67 Protein Immunodetection
Get PDFBACKGROUND: 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 cell
Advance-Platelet Rich Fibrin and Hyaluronic Acid Combination Improves Interleukin-6 and Granulation Index in Diabetic Foot Ulcer Patients
Get PDFBACKGROUND: 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
Comparison of Line Probe Assay (LPA) and Mycobacterium Growth Indicator Tubes (MGIT) Assay for Second-line TB Drug Susceptibility Testing
Get PDFBACKGROUND: Tuberculosis (TB) infection is one of the most prominent health issues in the world, including in Indonesia. TB is evolving into multidrug-resistant tuberculosis (MDR-TB) and requiring second-line TB drugs. Mycobacterium growth indicator tube (MGIT) is the gold standard for susceptibility testing of second-line TB drugs. Alternatively, line probe assay (LPA), which detects genes resistant to second-line TB drugs, takes a shorter time to run. This study aims to compare MGIT and LPA's ability to detect TB resistance to second-line TB drugs and observe mutation patterns of genes encoding second-line TB drugs.METHODS: This was an observational analytic study, using cross-sectional method. The data were acquired from the MDR-TB clinic’s medical records at the Dr. Hasan Sadikin Hospital from September to December 2019. LPA and MGIT test were conducted at the Health Laboratory Hall of West Java Province, then tested using Kolmogorov-Smirnov and chi-square statistic.RESULTS: From 121 subjects, 113 people were not resistant to the second-line TB drugs, which was examined using both LPA and MGIT (93.4%), p=0.991. Mutations were found in gyrA and rrs gene. There was no significant difference between the proportion of subjects resistant to the second-line of TB drugs tested using LPA and MGIT.CONCLUSION: LPA is an alternative method to MGIT because it requires a shorter time and reduces the risk of exposure that will improve MDR-TB patients management.KEYWORDS: line probe assay (LPA), multidrug-resistant TB, mycobacterium growth indicator tube (MGIT), second-line TB drugs
Simvastatin Improves Renal Function and Glomerulosclerosis in Ischemic-reperfusion Injury
Get PDFBACKGROUND: Statin is an anti-cholesterol drug that is widely prescribed throughout the world. Statins are mainly used to treat and prevent cardiovascular disease. Several studies have found the pleiotropic effect of statin. However, related effect of statin in kidney failure is still unclear. Ischemic-reperfusion (I/R) injury is a major cause of acute kidney failure. This study aims to determine the effect of simvastatin on kidney function and glomerular conditions by periodic acid-schiff staining in I/R injury.METHODS: Eighteen male Swiss mice were grouped into sham operation group (GSO), I/R injury group (GIRI), and simvastatin group (GSIM). The GSO group was performed by sham operation and pretreatment of 1% carboxymethylcellulose (CMC) for 3 days. The GIRI group was performed by I/R procedure and pretreatment of 1% CMC for 3 days and the GSIM group was performed by I/R procedure and pretreatment of 10 mg/kg BW simvastatin for 3 days. Blood urea nitrogen (BUN) and creatinine serum were assessed to determine kidney function. Histopathological analysis of glomerulosclerosis was assessed by the extent of glomerular damage (sclerosis), capillary loops, and synechia. The data were analyzed by one-way ANOVA followed by post hoc Tukey’s test (p<0.05).RESULTS: The creatinine and BUN levels in the GIRI group were the highest (0.97±0.48) compared with the other groups. The glomerulosclerosis index in the GSO group was 0.75±0.56, the GIRI group was 3.55±0.61, and the GSIM group was 2.08±1.37. There was a significant difference in the glomerulosclerosis index between the GSO and GIRI groups, but there was no significant difference between the GIRI and GSIM groups. These differences include the formation of sclerosis in the glomerulus, capillary loop, and synechiae.CONCLUSION: Simvastatin improves kidney function and glomerulosclerosis in I/R injury.KEYWORDS: ischemic-reperfusion injury, simvastatin, glomerulosclerosi
Administration of Vitamin D3 Improves Hemoglobin Level by Regulating TNF-a and IL-6 in DSS-induced Colitis Mice
Get PDFBACKGROUND: Anemia is frequently found in ulcerative colitis (UC) patients and assumed to be related to inflammatory process. Vitamin D3 (VD) is known to have anti-inflammatory and immunomodulatory effects. It also has the potential to be an alternative treatment of the inflammatory process that occurs at UC, however its mechanism has not been clearly established. This study aimed to assess the effect of VD on histopathology and hemoglobin levels in UC through its regulation in tumor necrosis factor (TNF)-α and interleukin (IL)-6.METHODS: Total samples of 24 mice were divided equally into Sham group, UC group, UC+VD group (given 3% dextran sodium sulfate (DSS) followed by VD), and VD+UC group (given VD followed by 3% DSS). Mouse Colitis Histology Index (MCHI) was used to measure histopathological changes. Immunohistochemical staining was used to observe expression of TNF-α and IL-6 in colon. Evaluation of anemia was determined by hemoglobin levels.RESULTS: Based on MCHI scores, significant epithelial damage was found in colon sample of UC group (8.25±3.05) compared to Sham (0.33±0.26), UC+VD (2.33±1.07), and VD+UC group (2.83±0.75) (p<0.05). Significant lower numbers of TNF-α were found in Sham (27.33±3.42), UC+VD (36.33±1.86), and VD+UC group (36.68±1.86) compared with UC group (44.66±4.87) (p<0.05). Significant less IL-6 expression was found in Sham (18.05±2.96), UC+VD (24.78±0.79), and VD+UC group (25.09±2.79) compared to UC group (38.85±3.51) (p<0.05). Differences in hemoglobin levels were significantly lower in UC group (11.85±0.97) compared to Sham (14.25±0.47), UC+VD (13.68±0.68), VD+UC group (13.52±1.07) (p<0.05).CONCLUSION: VD significantly reduced proinflammatory cytokines, increased mucosal repair, and improved hemoglobin levels.KEYWORDS: colitis, ulcerative, interleukin-6, tumor necrosis factor-alph
Effect of Croton caudatus Geiseler Aqueous Root Extract on Reproductive and Biochemical Parameters in Male Wistar Rats
Get PDFBACKGROUND: Croton caudatus Geiseler (CCG), a local herb that empirically has been used as traditional medicine for malaria, fever, convulsions, and rheumatic arthritis. This study aimed to investigate the effect of CCG on reproductive and biochemical parameters in male Wistar rats.METHODS: Twenty-four Wistar male rats were divided randomly into four groups, which were treated with CCG aqueous root extract at the concentration of 0-16 mg/kg, via oral gavage. After 21 days of treatment, blood, sperm, and testes were harvested for analyses. Sperm parameters were evaluated using computer-assisted sperm analyser (Hamilton Thorne Sperm Analyzer), and morphometric analyses of histological changes were performed.RESULTS: CCG extract at 8 and 16 mg/kg significantly (p<0.05) increased the total sperm count, concentration, and motility; whereby, only 16 mg/kg significantly decreased the percentage of abnormal sperm morphology. Testosterone hormones levels were the highest in 4 mg/kg CCG but were slightly decreased in 8 and 16 mg/kg CCG. The histological results showed a significant increase in the germinal epithelial height, decrease of lumen size at 16 mg/kg of CCG extract. No significant changes of alanine aminotransferase (ALT) and creatinine levels in serum from all CCG extract groups. However, the aspartate aminotransferase (AST) level in 8 mg/kg CCG was elevated, and the blood urea nitrogen (BUN) level was decreased in all CCG extract groups.CONCLUSION: In conclusion, the CCG extract at 4 mg/kg has the potential to enhance spermatogenesis activity within the seminiferous tubules in rats without any hepatotoxicity and nephrotoxicity effect on the liver and kidney.KEYWORDS: Croton caudatus Geiseler, rats, spermatogenesis, seminiferous tubules, testis, testosteron
Hypothalamic Microinflammation: New Paradigm In Obesity And Metabolic Disease
Get PDFBACKGROUND: Hypothalamus is the master regulator of body’s systemic homeostasis including energy balance, body temperature, sleep, blood pressure, and circadian rhythms. This review article will highlight the shifting of the old paradigm of obesityinflammation-metabolic syndrome, which was focused on visceral organs and systemic inflammation, into a new model involving microinflammation in the master regulator of endocrine system, i.e., hypothalamus.CONTENT: Since the early stage of over-nutritional conditions and aging process, microinflammation in hypothalamus has started to emerge, due to the activation of several proinflammatory signaling pathways, especially the nuclear factor kappa B (NF-kB) and c-Jun N-terminal kinase (JNK)-mediated nuclear transcriptional programs. Together with intracellular organelle stress signals, these pathways develop a chronic microinflammatory environment in the hypothalamus leading to obesity and metabolic disorders.SUMMARY: Hypothalamic inflammation has been noted not only as an important driver of impaired energy balance, but also contribute in altered neurocircuit functions and promote obesity-associated metabolic impairment.KEYWORDS: hypothalamus, inflammation, metabolism, obesity, metabolic syndrom
Immunohistochemical Expression of EGFR, NF-kB and Cyclin D1 in Sinonasal Inverted Papilloma and Squamous Cell Carcinoma
Get PDFBACKGROUND: Sinonasal inverted papilloma (SIP), a benign epithelial growth in the sinonasal region with epidermoid epithelial transformation, has been known for its invasiveness, recurrency, and its link with malignancy. Meanwhile sinonasal squamous cell carcinoma (SSCC) is an epithelial malignancy on squamous cells from the sinonasal region. Epidermal growth factor receptor (EGFR), Nuclear Factor kB (NF-kB), and Cyclin D1 are factors those might play important role in proliferation of SIP and SSCC. This research was conducted to investigate the expressions of EGFR, NF-kB and Cyclin D1 in SIP and SSCC.METHODS: A cross-sectional study by examining the EGFR, NF-kB, and Cyclin D1 immunohistochemical expressions of SIP and SSCC was conducted. Subjects whose blocks were used in this research, were diagnosed as SIP and SSCC at the Otorhinolaryngology-Head and Neck Surgery Clinic, Dr. Saiful Anwar General Hospital. Samples were selected, processed for inmmunohistochemistry, evaluated and statistical analyzed.RESULTS: Twenty-four SIP and 9 SSCC subjects with their paraffin blocks were selected. Clear immunohistochemical expressions of EGFR, NF-kB, and Cyclin D1 were observed for both SIP and SSCC. Significantly higher immunostaining levels of EGFR (45.6%, p=0.001) and NF-kB (42.2%, p=0.013) were observed in SSCC. Immunostaining levels of EGFR vs. NF-kB were moderately correlated (p=0.03, r=0.437), while the immunostaining levels of NF-kB vs. Cyclin D1 were strongly correlated (p=0.002, r=0.602).CONCLUSION: Expression of EGFR and NF-kB in SSCC were higher than the EGFR and NF-kB expression in SIP, suggesting that EGFR and NF-kB play important role in sinonasal malignancy.KEYWORDS: sinonasal, inverted papilloma, SCC, EGFR, NF-kB, Cyclin D