Molecular and Cellular Biomedical Sciences (E-Journal)
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Allogeneic Mesenchymal Stem Cells and Its Conditioned Medium as a Potential Adjuvant Therapy for COVID-19
No full textRecent research has demonstrated that mesenchymal stem cells (MSCs) potentially benefit and enhance coronavirus disease (COVID-19) recovery. This benefit occurs via a mechanism that promotes viral clearance by phagocytes and macrophages. This action occurs through the innate (increase in IL-10 production and decrease in TNF-α and IL-12 production) and the adaptive immune system (decrease in IL-17 production, promote regulatory T cell proliferation and inhibit effectors T cell proliferation). MSCs are expected to act as an anti-inflammatory in the hyper-inflammatory state of COVID-19. MSCs enhance immune cell replacement that have been overwhelmed or have been lost due to cytokine storm. Although vaccines are the answer to this pandemic, MSCs can improve COVID-19 patients, especially in patients with chronic illnesses. The focus on keeping death-rates low is a great opportunity for MSCs-based therapy for severe or critically ill patients. MSCs and conditioned medium have the potential to serve as adjunctive therapy in preventing the body's overactive defense response or the so-called cytokine storm caused by COVID-19.Keywords: adjuvant therapy, COVID-19, mesenchymal stem cells, secretom
The Role of Malondialdehyde (MDA) and Ferric Reducing Antioxidant Power (FRAP) in Patients with Hypertension
No full textOxidative stress contributes to the pathogenesis of hypertension and studies have shown that hypertension is associated with an increase in oxidative stress. Reactive oxygen species (ROS) lead to hypertension and antioxidants may be beneficial for its prevention. The main cause of oxidative stress in hypertension is endothelial dysfunction due to the malfunctions in the vasodilator systems, specifically the molecular mechanism of ROS and nitric oxide (NO). The level of malondialdehyde (MDA), a biomarker of lipid peroxidation and oxidative stress, is found to be higher in hypertension patients. Total antioxidant capacity (TAC), which has a strong relationship with blood pressure, is determined through the ferric reducing antioxidant power (FRAP). The aim of the review article is to elucidate the role of MDA and FRAP in hypertension.Keywords: oxidative stress, hypertension, blood pressure, oxidative damage, malondialdehyde, FRA
Association between Maternal FUT2 204A>G (rs492602) Genetic Polymorphism and Congenital Heart Disease in the Indian Population: A Study in Maternal-fetal Dyads
No full textBackground: FUT2 secretor genetic variants are strongly associated with absorption and circulatory levels of vitamin B12, thereby affecting folate metabolism pathway. The aim of this study was to evaluate the association between maternal FUT2 204A>G (rs492602) genetic polymorphism and CHD in the Indian population.Materials and method: One hundred and ten pregnant women who were vitamin B12 deficient with fetuses diagnosed with CHD were included in the case group and an equal number of healthy pregnant women with normal fetuses were selected as the control group. DNA was extracted from blood and umbilical cord tissue samples, and genotyped for FUT2 rs492602 polymorphism using allele-specific polymerase chain reaction. Hardy–Weinberg equilibrium test was used to calculate allele and genotype frequencies.Results: Significant increase in the frequency of AG (odds ratio=2.25; 95% CI: 1.25–4.05; p=0.009) and GG (odds ratio=3.51; 95% CI: 1.47-8.43; p=0.006) genotypes as well as G allele of FUT2 rs492602 were observed in the maternal case group. Furthermore, in the fetus case group, there was a significantly higher incidence of GG genotype (odds ratio=2.87; 95% CI: 1.26–6.57; p=0.018) and G allele (odds ratio=1.70; 95% CI: 1.15–2.53; p=0.009).Conclusion: FUT2 rs492602 are associated with CHD in the Indian population. Maternal genetic polymorphism that regulates vitamin B12 metabolic pathway might influence fetal cardiac development, thus serving as a predictor for CHD.Keywords: congenital heart disease, FUT2, single nucleotide polymorphism (SNP), vitamin B1
CRISPR Target-based Single-guide RNA (sgRNA) for Diagnostic Testing of Hepatitis B Virus
No full textBackground: Indonesia is the second-highest country with hepatitis B cases in the South East Asian region. Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 12 (Cas12) could be developed as a diagnostic tool to detect hepatitis B infection. This study was aimed to develop a diagnostic method for hepatitis B virus by designing CRISPR target-based single-guide RNA (sgRNA).Materials and method: The preCore/Core-gene sequences of hepatitis B virus were collected from the National Center for Biotechnology Information (NCBI) website. The selected sequence was submitted to Cas Designer and CRISPOR tools to design sgRNA. The resulting sgRNA was cloned in silico into an expression vector using Benchling software.Results: The 23-nucleotide sequence 5'- GTAGTCAGTTATGTCAATGTTAA-3’ had 30% GC content, 68.3 out-of-frame and 76 predicted efficiencies. This sequence had no mismatch based on analysis.Conclusion: This preliminary study will help design a CRISPR-based diagnostic kit for the detection of hepatitis B virus in Indonesia. However, further in vitro and in vivo studies are required to demonstrate its potential and efficiency.Keywords: CRISPR-Cas12b, diagnostic, HBV, sgRNA
Serum Copper, Iron, and Total Iron Binding Capacity in Hypothyroidism: A Case Control Study
No full textBackground: Thyroid hormone metabolism is linked to iron metabolism. Thyroperoxidase, a key enzyme in the biosynthesis of thyroid hormones, is iron-dependent. Thus, iron deficiency might be the primary cause of hypothyroidism. Copper is another trace element that has been linked to thyroid status. Copper regulates excessive thyroxine (T4) absorption and reduces cell damage during thyroid hormone synthesis. The present study clarified the possible correlations between iron and copper levels as well as total iron binding capacity (TIBC) and triiodothyronine (T3), T4, as well as thyroid-stimulating hormone (TSH) levels in healthy and hypothyroid subjects.Materials and methods: Thirty-five healthy subjects and 35 hypothyroid subjects were included in this study. Serum T3, T4, and TSH levels were measured using the enzyme linked fluorescence assay. Serum iron levels and TIBC were estimated using ferrozine/magnesium carbonate method, while serum copper levels were estimated using colorimetric method.Results: Copper levels were not significantly different between healthy and hypothyroid subjects. Iron and T4 levels were significantly lower in hypothyroid subjects compared with those in healthy subjects, while TIBC and TSH levels were significantly higher. There was no significant correlation between copper levels and T3, T4, and TSH levels.Conclusion: There were inverse correlations between TIBC and T4 as well as iron levels, and there was no significant correlation between copper levels and all thyroid function parameters. Routine examination of iron levels and thyroid function is highly recommended for early diagnosis and therapy of hypothyroidism.Keywords: total iron binding capacity, iron, copper, hypothyroidis
Genotype AA of ACE2 G8790A (rs2285666) Has Protective Potential Against COVID-19 Disease Severity
No full textBackground: SARS-CoV-2 virus uses angiotensin converting enzyme 2 (ACE2), a key enzyme of the renin angiotensin system (RAS) as the functional receptor for cell fusion and induction of infections in the respiratory system. Functional ACE2 gene polymorphisms may lead to RAS imbalance and are associated with COVID-19 susceptibility and severity. ACE2 G8790A (rs2285666), a splice region variant, is well characterized in various populations across the world. In the present study, the role of ACE2 G8790A (rs2285666) variant as risk predictor for severity of COVID-19 infection was investigated.Materials and methods: One-hundred COVID-19 subjects were included in the study and divided into: subjects with a history of severe infection and ICU-admitted (Group 1) and subjects with mild to moderate COVID-19 infection (Group 2). Genotype analysis for rs2285666 of ACE2 was performed using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method.Results: The distribution of ACE2 G8790A (rs2285666) genotypes were GG 62%, GA 18%, and AA 20% in Group 1 and GG 34%, GA 14%, and AA 52% in Group 2, respectively. The A allele of rs2285666 (p≤0.001; OR=3.4; 95% CI=1.89–6.107) were less frequent in Group 1 as compared to Group 2. Also, a statistically significant difference was found between severity of COVID-19 infection with age and comorbidities such as diabetes, hypertension, chronic kidney disease, but not gender.Conclusion: Our findings suggest the possibility of a protective mechanism of the AA genotype of ACE2 G8790A (rs2285666) variant against COVID-19 disease severity.Keywords: COVID-19, ACE2 gene, renin-angiotensin system, genetic association, rs2285666, sanger sequencin
Integrative Bioinformatics Reveals the Lactate Dehydrogenase B (LDHB) Significance in Colon Adenocarcinoma
No full textBackground: Lactate dehydrogenase B (LDHB), a typical oxidoreductase for converting lactate to pyruvate in the glycolysis process, takes a complex function in the progression of cancer cells. Even so, the profile of LDHB relevance in colon adenocarcinoma (COAD) remains ambiguous. Hence this study analyzed the expression and co-expression profile of LDHB, and its immune correlation in COAD.Materials and method: The mRNA expression and co-expression of LDHB in COAD were retrieved from UALCAN. The immune infiltration levels of LDHB from B cells, CD4+ T cells, CD8+ T cells, macrophages, neutrophils, and dendritic cells in COAD were assessed using the TIMER database. For assessing gene ontology and the KEGG pathway, DAVID v6.8 was utilized. The protein-protein interaction of LDHB-correlated genes was analyzed using STRINGDB and Cytoscape.Results: Significantly high expression of LDHB in COAD was spotted in several sample types and associated with a poor overall survival rate. Further, LDHB corresponded to the level of CD4+, macrophages, and myeloid-derived suppressor cell (MDSC) immune infiltrating cells. The co-expression of LDHB was associated with several essential genes for cell cycle progression.Conclusion: The findings of this study indicate an upcoming involvement of LDHB in COAD tumorigenesis and prognosis. Additionally, this study highlights the immune correlation of LDHB in COAD as preliminary data in developing diagnosis and treatment with a novel immune checkpoint in COAD.Keywords: lactate dehydrogenase, colon adenocarcinoma, expression, survival, immun
Neutralizing Antibody Response by Inactivated SARS-CoV-2 Vaccine on Healthcare Workers
No full textBackground: Currently, the key to combat coronavirus disease 2019 (COVID-19) as a global pandemic is relying mainly on vaccination, and several factors might affect the level of protection. This study aimed to determine the quantitative increase of neutralizing antibody titer against COVID-19 and the influence of gender, body mass index (BMI), routine consumption of vitamin C, D, and E towards the neutralizing antibodies after vaccination.Materials and methods: One hundred nine health workers from various health facilities were recruited. Sinovac inactivated severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) vaccine was used in this study. Antibody titer measurements were carried out quantitatively using electrochemiluminescence immunoassay (ECLIA) on day 14 after the first and second doses administration of the vaccine.Results: The average of antibody titers after the first and second doses were 109.1 and 191.6 U/mL, respectively. Antibody titer significantly increased (p=0.000) as much as 82.5 U/mL from the first to the second dose. There was a significant difference in the increase in antibody titer between respondents who consumed vitamin E regularly and those who did not (p=0.036). Routine consumption of vitamin C and D, gender, and BMI did not affect the increase in neutralizing antibody titer with p-values of 0.983, 0.337, 0.186, and 0.424, respectively.Conclusion: Routine consumption of vitamin E is associated with post-SARS-CoV-2 vaccination neutralizing antibody response. Gender, BMI, and the routine consumption of vitamin C and D have no association with the immune response.Keywords: COVID-19, neutralizing antibody, inactivated SARS-CoV-2 vaccin
Hypoglycaemic, Hypolipidaemic and Antioxidant Properties of Celastrus paniculatus Seed Extract in STZ-induced Diabetic Rats
No full textBackground: Celastrus paniculatus is a herb used in the Ayurvedic system of medicine that has been reported to show multiple pharmacological properties. In this study, we explored the antioxidative, hypolipidaemic and hypoglycaemic potential of C. paniculatus methanolic seed extract (CPMSE) in high-fat diet (HFD)/streptozotocin (STZ)-induced type 2 diabetes mellitus (T2DM) rats.Materials and methods: Seeds of C. paniculatus were extracted in methanol using Soxhlet extraction method. A total of 36 rats were induced with STZ and HFD and treated with glibenclamide or various concentrations of CPMSE. Upon treatment, blood samples were collected and kidney and liver samples were homogenised. Serum biochemical estimation was performed using several diagnostic kits. Protein was estimated by bicinchoninic acid (BCA) method. Oxidative stress was assessed by measuring malondialdehyde level and superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST) activity.Results: CPMSE caused improvements in glucose homeostasis, lipid profile, liver function and oxidative stress parameters in a dose-dependent manner. CPMSE significantly decreased the levels of fasting blood glucose and glycated haemoglobin as well as increased insulin level and total protein content. There was an increase in total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), triglycerides (TG) levels and reduction in high density lipoprotein-cholesterol (HDL-C) level. There was a decrease in serum levels of serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT) and alkaline phosphatase (ALP). CPMSE decreased LPO and increased CAT, SOD and GST activity.Conclusion: CPMSE has hypoglycaemic, hypolipidaemic and antioxidant properties by reducing the oxidative stress.Keywords: diabetes mellitus, Celastrus paniculatus, antioxidant, phytochemicals, phytonutrients, streptozotocin, high-fat die
The Construction of A Multi-epitope Vaccine Against Klebsiella pneumoniae Using in silico Approach
No full textBackground: Klebsiella pneumoniae is one of the bacteria that causes pneumonia infection. Even though the number of pneumonia cases is relatively high and has become a global problem, there is still no vaccine available to prevent this disease. This study was aimed to design a multi-epitope vaccine design through an in silico approach, against K. pneumoniae.Materials and method: Vaccine candidate was constructed based on proteins of K. pneumoniae. These proteins were analyzed to identify the antigens sequence for multi-epitope vaccine design. The constructed vaccine was predicted for allergenicity, toxicity, population coverage, and its physicochemical properties. The vaccine structure was then docked with the toll like receptor 2 (TLR2) molecule to show the interaction. Expression analysis and cloning of the constructed vaccine was carried out in the pET-28a vector using SnapGene.Results: The vaccine was 567 amino acids long, consisting of Cholera Toxin Subunit B as an adjuvant, 6 B-cell epitopes, 11 cytotoxic T-cell epitopes, and 10 helper T-cell epitopes connected with the appropriate linker. Epitopes analysis showed that the vaccine will be a non-toxic, has high antigenicity, but non-allergenic. The vaccine was predicted to be stable, hydrophilic, and had a low risk of triggering autoimmune response. The vaccine molecule was compatible to humans TLR2 molecule. Furthermore, visualization of the candidate vaccine protein on pET-28a showed that the vaccine protein might be expressed correctly.Conclusion: The construction of multi-epitope vaccine has been developed, which might be a good vaccine candidate, containing 6 B-cell epitopes, 11 CTL epitopes, and 10 HTL epitopes. The construct may help scientists to experimentally formulate multi-epitope vaccine against K. pneumoniae in the future.Keywords: in silico, Klebsiella pneumoniae, multi-epitope, vaccine