Molecular and Cellular Biomedical Sciences (E-Journal)
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Correlation between Genetic Polymorphism of CYP2A13 Genotype and Lung Cancer in Female Passive Smokers
No full textBackground: Nicotine is metabolized to cotinine by cytochrome P450 enzyme, and this enzyme is involved in the activation of toxic and carcinogenic substances. The aim of this research was to assess the relationship between genetic polymorphism of CYP2A13 and lung cancer incidence in female passive smokers.Materials and methods: This research was a case-control study that involved 104 research subjects. Subjects were recruited through purposive sampling technique from 2 hospitals in Medan, North Sumatra, Indonesia. The case population consisted of female passive smokers with lung cancer and the control population consisted of female passive smokers without lung cancer. All research subjects underwent blood sampling for genomics DNA extraction and CYP2A13 genotyping by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Data was analyzed by conditional logistic regression by Epi Info 7.0 software.Results: Among 104 subjects, 26 (25%) individuals were heterozygous, 76 (73%) individuals were wild type, and 2 (2%) were mutant for the 257Cys allele. There was a significant correlation between CYP2A13 genotype and lung cancer incidence (p-value<0.05). Female passive smokers with CT genotype had 2.7 greater risk of developing lung cancer than those with CC genotype (wild type). The C allele had more frequency and 1.6 times higher risk of lung cancer compared to T allele with a wide confidence range (0.73–3.52).Conclusion: There was a significant correlation between CYP2A13 polymorphism and lung cancer incidence in female passive smokers.Keywords: polymorphism, CYP2A13, PCR-RFLP, female passive smoker, lung cance
ZEB1 is Negatively Correlated with E-Cadherin in Prostatic Anomaly Tissue
No full textBackground: Prostatic anomalies are common in tumor or infection condition. The enlargement of prostate gland affects the epithelial cell polarity that involves epithelial-mesenchymal transition (EMT). Transition into mesenchymal is mediated by transcription factor ZEB1 and E-cadherin protein. Upregulation of ZEB1 and loss of E-Cadherin expression were associated to proliferation and metastasis of malignancy cells. This study aims to describe the correlation of ZEB1 and E-cadherin expression in prostatic anomaly.Materials and method: Samples were Formalin Fixed Paraffin Embedded (FFPE) block consist of 8 block Benign Prostatic Hyperplasia (BPH), 6 blocks High Grade Prostatic Intraepithelial Neoplasia (HGPIN) and 6 blocks Prostate Carcinoma (PCA). The blocks then sliced into 5 sections to be prepared for RNA extraction procedures. ZEB1 and E-Cadherin expression was analyzed by semi-quantitative procedures using PCR and electrophoresis. Correlation between ZEB1 and E-Cadherin espression was analyzed using Spearman’s rank correlation.Results: Relative expression of ZEB1 and E-cadherin mRNA in each group of prostatic anomaly were not significantly different (p>0.05). ZEB1 and E-Cadherin mRNA expression showed a significant and moderate level of negative correlation (p<0.05; 0.40 < r < 0.59). Increasing of ZEB1 mRNA expression will be followed by decreasing of E-Cadherin mRNA expression.Conclusion: ZEB1 negatively correlates with E-cadherin due to EMT process in prostatic anomaly. High expression of ZEB1 induced down-regulation of E-cadherin and vise versa. Various studies can be developed, especially the development of targeted therapy against ZEB1 to suppress the EMT process by increasing the expression of E-cadherin.Keywords: epithelial-mesenchymal transition (EMT), ZEB1, E-Cadherin, BPH, HGPIN, PC
D-dimer as a Potential Biomarker of Severity in Children Confirmed with COVID-19
No full textBackground: Coronavirus disease 2019 (COVID-19) in children spreads easily and has a relatively high incidence. Severe complications in children confirmed with COVID-19 are thought to be related to the multisystem inflammatory syndrome, which is associated with coagulation disorders. D-dimer is a fibrin degradation end product which is easy to examine, affordable, fast and reliable. This study investigated the potency of D-dimer levels as a biomarker and assessed optimal cut-off value of D-dimer on severity of COVID-19 in children. Materials and methods: An analytical observational study with a cross-sectional design was conducted in children aged 1-18 years confirmed to have mild, moderate or severe COVID-19 who were treated in the isolation room of Dr. Moewardi Hospital, Surakarta, Indonesia from September 2021 to February 2022. Statistical analysis was conducted using Mann- Whitney test and p<0.05 was considered as statistically significant. The cut-off value of D-dimer was determined with the receiver operating characteristic (ROC) curve. Results: There were 39 children with COVID-19. They were in mild (n=14; 35.9%), moderate (n=19; 48.7%) and severe (n= 6; 15.4%) stages. There were significant differences in D-dimer levels between mild and moderate stages (p=0.001), and mild and severe stages (p=0.001). No significant difference in D-dimer levels between moderate and severe stages (p=0.162). The cut-off value of D-dimer was 485 μg/mL with 92% sensitivity and 71.4% specificity. Conclusion: D-dimer can be used as a potential biomarker of severity in children with COVID-19.Keywords: D-dimer, COVID-19, severity, childre
Association of CYP2A6 Genetic Polymorphism and Lung Cancer in Female Never Smokers
No full textBackground: The major significant factor that affected lung cancer development among female passive smokers is environmental tobacco smoke. Nicotine can be found in a never smoker population, such as a child whose father is a smoker. Lung carcinogenesis in never smoker populations is affected by nicotine metabolism by CYP2A6 gene, which encodes the main nicotine metabolizing-enzyme. The aim of this study was to assess the genetic polymorphism of CYP2A6 and its association with secondhand smokers among females who have suffered from lung cancer in North Sumatra population.Materials and methods: This study was a case-control study, composed of 53 case subjects and 46 control subjects that were involved through a purposive sampling technique from two hospitals in Medan. PCR-RFLP was used for the examination of CYP2A6 gene to determine the genotype. The data were analyzed with conditional logistic regression test using Epi Info 7.0 software.Results: The most common genotype of CYP2A6 detected in this study was *1B/*1B (40.4%), while *1B allele had the highest prevalence (55.5%). There was no significant association between CYP2A6 genotype (p-value=0.61) or alleles (p-value=0.25) and the incidence of lung cancer.Conclusion: There was no association between CYP2A6 polymorphism and the incidence of lung cancer in secondhand smoker females.Keywords: CYP2A6, PCR-RFLP, female secondhand smokers, lung cance
Genetic Variant of Vascular Endothelial Growth Factor (VEGF)-A rs699947 is Associated with Preeclampsia
No full textBackground: Preeclampsia remains as the leading cause of maternal-neonatal mortality and morbidity worldwide. Vascular endothelial growth factor A (VEGF-A) is a proangiogenic factor related to endothelial dysfunction and plays an important role in the preeclampsia pathophysiology. Genetic variants of VEGF-A are associated with VEGF-A expression and preeclampsia risk, however there are still inconsistent results between different populations. The aim of this study was to determine the association of this genetic variant as preeclampsia risk factor.Materials and methods: A cross-sectional study was performed with 76 pregnant women (29 preeclampsia and 47 normotensive) Jambi-Malay ethnic subjects. Sample DNA was extracted from subject’s blood. To determine the genotype, one-step tetra amplification refractory mutation system (ARMS) polymerase chain reaction (PCR) method for VEGF-A rs699947 C/A was used.Results: We found that pregnant woman with AC genotype (p-value=0.045; OR=2.76 ; 95% CI=1.01-7.58) and AA genotype (p-value=0.026; OR=12.44; 95% CI=1.23-126.18) had higher risk of preeclampsia than the CC genotype.Conclusion: Genetic variant VEGF-A rs699947 C/A is associated with preeclampsia. The AC and AA genotype is the risk genotype for preeclampsia in Jambi-Malay ethnics.Keywords: preeclampsia, VEGF-A, genetic variant, Jambi-Malay, Indonesi
Utilization of Expired Platelet Concentrate for Production of Human Platelet Lysate as a Medium for T47D Cell Propagation
No full textBackground: Platelet concentrate (PC) has a short shelf life (5 days). Expired PC cannot be used for clinical purposes. PC is used for human platelet lysate (HPL) production, which was found to be more effective than FBS at increasing T47D cell proliferation. HPL production using expired PC has not been reported. This study aimed to investigate whether the use of HPL produced from expired PC (storage duration >5 days) can increase the proliferation of T47D cells in vitro.Materials and methods: Expired PC samples with a shelf life of 7 and 11 days were used to produce HPL via freeze/thaw method. pH, total protein content, glucose and albumin levels were measured. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure proliferation rate and doubling time of HPL-treated T47D cells.Results: After HPL production, the glucose level was influenced by the pH (p=0.003), and albumin level was influenced by total protein content (p=0.030). HPL stored for 7 and 11 days increased cell proliferation rate by 1.41 and 1.80 times higher than 10% FBS, respectively. HPL produced from expired PC did not cause morphological abnormality of the cells. In this study, the glucose levels affected cell proliferation (p=0.030). High glucose levels inhibited T47D cell proliferation.Conclusion: Expired PC can be used as a potential material for HPL production, since HPL produced from expired PC increases cell proliferation rate and shortens cell doubling time.Keywords: cell proliferation, human platelet lysate, platelet concentrate, thrombocyte, T47
Safety and Efficacy of Mesenchymal Stem Cells in Burn Therapy: Systematic Review
No full textThe experimental research on the use of mesenchymal stem cells (MSCs) for burn therapy has been published several times. However, current clinical procedure remains a challenging discussion. This systematic review assesses the safety and efficacy of administering mesenchymal stem cells (MSCs) to burns and determines the most effective source of MSCs for burn therapy. We reviewed several studies through PubMed, Google Scholar, Science Direct, and DOAJ online databases. PRISMA-P 2020 method was used based on inclusion and exclusion criteria that were re-selected through Joanna Briggs Institute (JBI) Critical Appraisal Tools. Results from 13 articles showed that MSCs are safe for burn therapy with minimal side effects/complications and have the potential to repair tissue and accelerate burn healing through several mechanisms. The treatment of MSCs in burns is influenced by donor characteristics and related to the severity and area of the burn. It can be concluded that the administration of MSCs is safe and effective in burn therapy. Keywords: burns, mesenchymal stem cells, therapeutic safety, therapeutic efficacy, wound healin
Non-Synonymous Mutation Analysis of SARS-CoV-2 ORF3a in Indonesia
No full textBackground: The report of mutation sites ORF3a SARS CoV-2 in Indonesia is still limited. Some research showed that mutations in ORF3a protein might alter SARS-CoV-2 pathogenesis. Observation of new variants should be conducted as a risk monitoring framework.Materials and method: We assessed the impact of mutations in ORF3a protein by analyzing 3,751 SARS-CoV-2 DNA sequences from the GISAID database from March 2020 until July 2021. The whole-genome sequences were aligned using Clustal Omega Multiple Sequence Alignment from EMBL-EBI and analyzed using BioEdit version 7.2.5 software. The reference whole genome sequence was taken from the Genbank database with accession number NC045512. We excluded the samples containing N letters due to inaccurate reading. Effect of point mutations on protein structure was analyzed using PredictProtein (https://predictprotein.org) and Protein Variation Effect Analyzer (PROVEAN) v1.1.3. online software.Results: We identified five most frequent non-synonymous mutations in ORF3a protein of SARS-CoV-2 which were Q57H (58.04%), S26L (27.25%), S220I (10.37%), D155H (8.98%), and P104S (5.47%).Conclusion: These mutation data showed the phenomenon of amino acid changes in ORF3a SARS-CoV-2 in Indonesia until July 2021. The implication of this mutation needs to be determined in further studies.Keywords: Indonesia, mutations, non-synonymous, SARS-CoV-2, whole genome
The Effects of Moringa oleifera Leaves on Complete Blood Count, Renal and Liver Functions as Potential Therapy for Malnutrition
No full textBackground: Moringa oleifera which is available in many areas all over the world including Sudan is low-cost and traditionally used in the treatment of many disorders, including malnutrition. This study aimed to determine the effect of aqueous extract of M. oleifera leaves in renal, liver functions and complete blood count (CBC) parameters, and its potential as therapy for malnutrition.Materials and methods: This was an experimental case control study using twenty-five Wistar albino rats. Rats were divided into three groups: normal protein diet group, low protein diet with or without M. oleifera extract groups. We determined rats' weight, CBC parameters, blood mineral concentrations, as well as liver and renal functions at day 0, 7, and 14.Results: Our findings showed that rats' weight were significantly different between the three groups at day 0, 7, and 14. Rats' weight, blood sodium, potassium, calcium, and urea concentration, as well as Hb concentration, TWBCs count, total platelets count, and %lymphocyte showed significant differences between three groups at day 0, 7, and 14.Conclusion: M. oleifera leaves can be used as potential therapy for malnutrition because they have some effects on weight, blood mineral concentrations, renal and liver function, as well as CBC parameters.Keywords: ALP, AST, ALT, creatinine, Moringa oleifer
Methanol Extract of Katuk (Sauropus androgynus) Leaves as an Anti-inflammatory Agent: Animal Study in Carrageenan-induced Rat Models of Inflammation
No full textBackground: Inflammation is a response in the human body to survive during infection, injury and tissue damage. Acute inflammation causes edema and polymorphonuclear neutrophils (PMNs) release. Sauropus androgynus leaves which contain flavonoids, tannins, saponins, steroids and triterpenoids may have anti-inflammatory properties. These compounds can be extracted with methanol. This research aimed to determine the anti-inflammatory effect of S. androgynus leaves methanol extract (SALME) on carrageenan induced-rats. Materials and methods: True experimental study was conducted using 12 Wistar rats. Rats were induced with carrageenan subcutaneously on the plantar pedis. Carrageenan-induced rats were treated with/without various doses of SALME. Edema volume was measured with a plethysmometer. The plantar pedis tissues were collected and stained with haematoxylin- eosin (HE) staining, then PMNs were observed and counted under a light microscope. All data were analyzed by one-way ANOVA, Bonferroni post hoc test, Pearson correlation and linear regression. Results: SALME had significant effects on the volume of edema (p=0.000) and the number of PMNs (p=0.000). The most effective dose to reduce the edema volume and decrease the PMNs cell number was 37.80 mg/200 g body weight (BW). SALME doses may affect 91.0% of edema volume and 89.2% of PMNs cell number. Edema volume had a significant, robust correlation (92.3%) with PMNs cell number. Conclusion: SALME is confirmed to have an anti-inflammatory activity by reducing the edema volume and decreasing the PMNs cell number. Keywords: Sauropus androgynus, methanol extract, anti-inflamamatory, carrageenan, plantar pedis, rat