Knowledge and perception regarding prenatal genetic screening of thalassemia, down syndrome and neural tube defects: a study among IIUM Kuantan undergraduates

Prenatal genetic screening is offered during pregnancy to detect foetuses that have certain diseases. It is widely used in the detection of congenital malformation which results in foetal birth defects. Unawareness of the society on the importance of prenatal genetic testing contributes to the increase in the birth defect rate.Future parents should be exposed with the importance in performing prenatal genetic screening.The purpose of this study was to examine the knowledge and perception level of International Islamic University Malaysia (IIUM) Kuantan students regarding prenatal genetic screening thalassemia, Down syndrome and neural tube defects.This is a cross-sectional study whereby192 respondents were selected using convenience sampling method. A set of close-ended questionnaire was distributed among students in IIUM Kuantan. Independent t-test, parametric test (One- Way ANOVA test), non-parametric test (Mann-Whitney test) and correlation coefficient (Pearson) were used to find all related factors influencing knowledge and perception and to find association between knowledge and perception of IIUM Kuantan students.From this study, it was found that the level of knowledge and perception of IIUM Kuantan students regarding prenatal genetic screening of thalassemia, Down syndrome and neural tube defects was relatively high. Married students hada betterknowledge compared to unmarriedstudents (p=0.008). Moreover, students from Kulliyyah (Faculty) of Medicine had adequate level of knowledge (p<0.001) and good perception (p=0.003) compared to students from other kulliyyahs. Age and year of study weresignificantly associated with students’ knowledge (p<0.001). In addition, ageand year of study were significantly associated with students’ perception(p=0.03 and p=0.007, respectively). The findings of this study depicted that adequate level of knowledge has ledto a good perception towards prenatal genetic screening among IIUM undergraduate students (p<0.001). Keywords: Prenatal genetic screening, undergraduate students, knowledge, perception, Malaysia

Non-invasive prenatal testing using cell-free fetal DNA from maternal plasma: a review

Nowadays, the use of cell-free fetal DNA (cfDNA) is a promising tool in clinical practice as a potential non-invasive prenatal testing (NIPT) method since it was discovered in early 1970s. The fetal DNA is approximately ~10% in a mixture of maternal DNA from maternal plasma and this amount will increase as gestation period increases. In recent years, the development of robust molecular analysis of fetal DNA namely via RT-PCR, next generation sequencing (NGS), digital PCR and massively parallel sequencing (MPS) helps the implementation of NIPT especially in fetal chromosomal aneuploidy detection. Thus, these analyses provide the alternative to the conventional invasive prenatal testing such as amniocentesis and chorionic villus sampling (CVS). The common fetal aneuploidy is trisomy 21 (T21) which is caused by an extra copy of all or part of chromosome 21 and known to be validated by amniocentesis approach as the gold standard method. Currently, the epigenetic detection of trisomy 21 had been introduced as a new non-invasive method that investigates the association between DNA methylation and gene expression in T21 fetal DNA. This review briefly summarizes the NIPT and invasive prenatal testing of fetal aneuploidy and recent molecular analysis study by using cfDNA from maternal plasma

Genetic and maternal factors in Hyperemesis Gravidarum: a systematic review

Hyperemesis gravidarum (HG) is a severe form of nausea and vomiting during pregnancy (NVP), which can lead to extreme dehydration, significant weight loss, and electrolyte and metabolic imbalances. Importantly, early identification of HG symptoms can help to reduce its severity and prevent complications. Although HG is associated with many adverse maternal and fetal outcomes, there is limited understanding of the risk factors. This review provides current data on genetic and maternal factors that are linked to HG. All observational studies published in English that investigated the genetic or maternal factors associated with HG from 2011 until 2021 were systematically searched using the PubMed, Scopus, and ProQuest electronic databases. A total of 1462 citation titles was identified, of which 47 potentially relevant abstracts were screened. Of those, 15 studies met the inclusion and exclusion criteria. The genetic variants in the ryanodine receptor 2 gene (RYR2), growth differentiation factor-15 (GDF15), and protein-coding insulin-like growth factor-binding protein 7 (IGFBP7) were found to be associated with HG. On the other hand, several potential maternal factors contributing to the onset of HG were age, Helicobacter pylori infection, body mass index (BMI) status, a history of HG in a previous pregnancy, carrying a female fetus, high serotonin levels, and reproductive factors. In view of the lack of strength of the overall evidence for risk factors related to HG, it is first imperative to establish a precise definition for HG in a diverse study population. Nevertheless, to conclude, this review was able to provide current data on genetic and maternal factors that are associated with HG

MGMT and SPOCK2 promoter methylation in diffuse large B-Cell lymphoma: a study in two tertiary health centres in the East Coast of Malaysia

MGMT (O6-Methylguanine-DNA Methyltransferase) suppresses tumor development by removing alkyl adduct, while SPOCK2 (SPARC/Osteonectin CWCV and Kazal-like domains proteoglycan) abolishes the inhibition of membrane-type matrix metalloproteinases (MT-MMP) which leads to angiogenesis. Hence, MGMT methylation may initiate malignant cells transformation. In contrast, SPOCK2 methylation is hypothesized not to be a common event in diffuse large B-cell lymphoma (DLBCL). In this study, we examined the methylation status of MGMT and SPOCK2 in DLBCL as in Malaysia the information is extremely lacking. A total of 88 formalin-fixed paraffin-embedded tissue of patients diagnosed with DLBCL from the year 2006 to 2013 were retrieved from Hospital Universiti Sains Malaysia, Kelantan and Hospital Tengku Ampuan Afzan, Pahang. Methylation-specific polymerase chain reaction (MSP) was used to examine the methylation status of both genes. Interestingly, methylation of MGMT was detected in all the 88 DLBCL samples, whereas SPOCK2 was found to be methylated in 83 of 88 (94.3%) DLBCL cases. Our study showed a remarkably high percentage of promoter methylation of both MGMT and SPOCK2 genes. Our finding also negates initial expectation that SPOCK2 methylation would be an uncommon event in the majority of DLBCL cases. This study has shown a very high percentage of promoter methylation of MGMT and SPOCK2 in the DLBCL cases studied by MSP, using archival lymphoma tissues. Nonetheless, additional research is needed to quantitatively evaluate MGMT and SPOCK2 methylation, and to analyse gene expression and/or protein expression in order to further understand the role of MGMT and SPOCK2 methylation in the pathogenesis of DLBCL

Methylation-specific PCR Revealed Aberrant Promoter Gene Methylation of p16, MGMT and SPOCK2 in Diffuse Large B-cell Lymphoma

DNA methylation silences the gene through addition of methyl group. p16, a tumor suppressor gene that inhibits cyclin-dependent kinase, inactivates the Rb protein and blocks G1 phase in a normal cell cycle. A DNA repair gene, MGMT removes alkyl adduct to a cysteine residue within the protein, thus preventing lethal cross-links. p16 and MGMT methylation has been reported to associate with DLBCL. A member of the extracellular chondroitin and heparin sulfate proteoglycans, SPOCK2 functions mainly in extracellular matrix for cell adhesion. Uniquely, SPOCK2 (testican 2) abolishes the inhibition of membrane-type 1-matrix metalloproteinase by other testican family which might enhance the angiogenesis. In this study, we aimed to screen for aberrantly methylated genes which might contribute to the pathogenesis of DLBCL using methylation specific PCR (MSP). p16 methylation was identified in 64 (73%) of 88 samples. On the other hand, SPOCK2 was found to be unmethylated in 30 (34%) samples. Interestingly, MGMT methylation was detected in all cases. We also found an association between p16 methylation status with patients aged >50 years old (p= 0.023). This finding is parallel with an animal study showing that aging increases p16 methylation. No association was found between the methylation of other genes with age. Unmethylation of SPOCK2 might cause testican 2 expressions, which has been suggested to contribute toward malignant behaviour. MGMT was reported to be methylated among cancer patients who smoke, drink and are non-vegetarian. Thus, it is hypothesized that lifestyle might affect MGMT methylation in this study population

Pyrosequencing-based quantitative identification of p16 methylation in diffuse large b-cell lymphoma at two centres in the east coast of malaysia

Introduction: Methylation of promoter region of p16 leading to gene silencing has been implicated in a wide range of malignancies including lymphomas. In diffuse large B cell lymphoma (DLBCL) particularly, a varying percentage of epigenetic inactivation of p16 promoter region was observed ranging from 16 - 54%. However, quantitative analysis of p16 promoter methylation in DLBCL has not been extensively studied in Malaysia. Objective: This study aims to quantitatively analyse p16 methylation in DLBCL samples using pyrosequencing technique. Methods: Genomic DNA was extracted from 16 formalin-fixed paraffin-embedded lymphoma tissue blocks from patients diagnosed with DLBCL. Samples were retrieved from Hospital Tengku Ampuan Afzan, Pahang and Hospital Universiti Sains Malaysia, Kelantan. Primers were designed to amplify bisulfite-treated DNA targeting p16 promoter region. Methylation status of 7 CpG sites was determined by pyrosequencing. Results: All the 16 samples studied showed promoter methylation of p16. The range of mean methylation percentage was between 18 to 81%. Conclusion: The present study has successfully measured the level of methylation of p16 in all 7 CpG sites despite the limitation in sample size. Since p16 methylation is a common event in our series of DLBCL cases, it is worth including a larger sample size in future studies to increase the chance of finding a significant correlation with clinical parameters

Association of Loss of Heterozygosity and PTEN Gene Abnormalities with Paraclinical, Clinical Modalities and Survival Time of Glioma Patients in Malaysia

BackgroundThe pattern of allelic loss of heterozygosity (LOH) and PTEN mutations appear to be associated with the progression of gliomas leading to a decrement in the survival rate of patients. This present study was carried out to determine the LOH and PTEN mutational status in glioma patients and its association with patients' survival.MethodsThirty-seven Malaysian glioma patients of the Malay race were subject to PTEN mutational analysis and the presence of LOH using the cold single-strand conformation polymorphism method, and their clinical and paraclinical response were correlated.ResultsAmong analysed glioma patients, seven (21.6%) cases with PTEN mutations were detected and 12 (32.4%) of 37 patients showed presence of LOH. Univariate analysis showed that tumour grade, vascularization, PTEN mutation, LOH and combination of both PTEN mutation and LOH were significantly associated with glioma patients' survival. Multivariate analysis revealed that no factors contributed to survival time.ConclusionThe results show that PTEN mutation and LOH are quite frequent in Malaysian glioma patients. However, they have no impact on the survival outcome of patients

Antibody response against Severe Acute Respiratory Syndrom Coronavirus 2 messenger Ribonucleic Acid vaccines in infected individuals: a systematic review

Individuals with a history of coronavirus disease 2019 (COVID-19) exhibit memory immunity acquired during natural infection. However, a decline in immunity after infection renders these individuals vulnerable to re-infection, in addition to a higher risk of infection with new variants. This systematic review examined related studies to elucidate the antibody response in these infected individuals after messenger ribonucleic acid (mRNA) vaccination. Hence, the focus of this review was to ascertain differences in the concentration of binding and neutralising antibodies of previously infected individuals in comparison to those of infection-naïve individuals after administration of two doses of mRNA vaccination through available case-control and cohort studies. Positive reverse transcriptase-polymerase chain reaction test or detectable anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies at the baseline in included studies showed categorisation of infected and uninfected individuals. This review utilised three online databases: PubMed, Scopus and Cochrane with the following keywords: (COVID-19 OR ‘Coronavirus Disease 2019’ OR SARS-CoV-2) AND Immun* AND (Pfizer OR BioNTech OR BNT162b2 OR Comirnaty OR Moderna OR mRNA-1273) from January 2019 to July 2021. Following the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocol (PRISMA-P) 2020 guidelines and assessment based on the Crowe Critical Appraisal Tool (CCAT), we included 13 related qualified papers of observational studies discerning the binding and neutralising antibody concentrations of infected and uninfected individuals after administration of mRNA vaccines, such as the BNT162b2 and mRNA-1273 vaccine. The mRNA vaccines induced robust binding and neutralising antibody responses in both groups. However, infected individuals showed induction of higher antibody responses in a shorter time compared to uninfected individuals. Hence, a single dose of mRNA vaccination for infected individuals may be sufficient to reach the same level of antibody concentration as that observed in uninfected individuals after receiving two doses of vaccination

p16 tumor suppressor gene methylation in diffuse large B cell lymphoma: a study of 88 cases at two hospitals in the East Coast of Malaysia

Introduction: p16 gene plays an important role in the normal cell cycle regulation. Methylation of p16 has been reported to be one of the epigenetic events contributing to the pathogenesis of diffuse large B-cell lymphoma (DLBCL) which occurring at varying frequency. DLBCL is an aggressive and high-grade malignancy which accounts for approximately 30% of all non-Hodgkin lymphoma cases. However, little is known regarding the epigenetic alterations of p16 gene in DLBCL cases in Malaysia. Therefore, the objective of this study was to examine the status of p16 methylation in DLBCL. Methods: A total of 88 formalin-fixed paraffin-embedded DLBCL tissues retrieved from two hospitals located in the east coast of Malaysia, namely Hospital Tengku Ampuan Afzan (HTAA) Pahang and Hospital Universiti Sains Malaysia (HUSM) Kelantan, were chosen for this study. DNA specimens were isolated and subsequently subjected to bisulfite treatment prior to methylation specific-PCR. Two pairs of primers were used to amplify methylated and unmethylated regions of p16 gene. The PCR products were then separated using agarose gel electrophoresis and visualised under UV illumination. SPSS version 12.0 was utilised to perform all statistical analysis. Result: p16 methylation was detected in 65 of 88 (74%) samples. There was a significant association between p16 methylation status and patients aged >50 years old (p=0.04). Conclusion: Our study demonstrated that methylation of p16 tumor suppressor gene in our DLBCL cases is common and significantly increased among patients aged 50 years and above. Aging is known to be an important risk factor in the development of cancers and we speculate that this might be due to the increased transformation of malignant cells in aging cell population. However, this has yet to be confirmed with further research and correlate the findings with clinicopathological parameters