Downregulation of WNT8A, LRP5, LRP6 and FGF8 in Malay non-syndromic cleft lip and/or palate patients

Non-syndromic cleft lip and/or palate (NSCL/P) is a one of the most common birth defects occurs as a result of multi-factorial determinants such as genetic and environmental factors. Genetic factor has been studying widely across different population in identifying genes causing cleft defects. This study aims to validate the role of fibroblast growth factors (FGFs) and signalling molecules Wingless-type (Wnt) to the occurrence of cleft lip and/or palate among Malay population. Tissue skin was obtained from consented NSCL/P patients who underwent the cleft lip repair operation at the upper lip skin area and non NSCL/P individual was obtained from patient having traumatic injury at the lip area as a control. Expression pattern of FGF8, FGF10, Wnt8a, and LRP5/6 were tested and validated using Western Blot (WB) and quantitative Reverse Transcriptase-PCR (qRT-PCR). The fold change difference of FGF8 (0.614 ± 0.1012-fold), FGF10 (0.7188 ± 0.1017-fold) and Wnt8a protein (0.9051 ± 0.0142-fold) was downregulated by 1-fold in cleft lip (CL) tissues compared to the normal meanwhile LRP5/6 protein (1.2201 ± 0.1404-fold) was found upregulated by 1-fold. Validation testing confirmed that expression of FGF8 (p=0.014), Wnt8a (0.0762 ± 0.0227), LRP5 (0.3577 ± 0.1362) and LRP6 (0.3093 ± 0.2541) were significantly reduced in CL tissues compared to normal. This is the first study identified defective regulation of Wnt8a, LRP5, LRP6 and FGF8 in NSCL/P among Malays. These novel findings clearly explained the important role of growth factors FGF and Wnt signalling pathway in lip and palate formation during craniofacial development

Skin Tissue Surface Morphology and Quality of RNA and Protein Extracted from Fresh and Stabilized Human Cleft Lip and Palate Tissue

Cleft lip palate is a human congenital disorder worldwide and the study of this genetic disease requires molecular genetic analysis. This analysis required the use of nucleic acid and protein, thus good quality and quantity of its extraction is important. We are comparing RNA and protein extractions from tissue biopsy of cleft lip palate in both fresh and stabilized condition. Tissue morphology was also captured using Scanning Electron Microscopy (SEM) for any morphology differences. Tissue homogenization may destroy nucleic acid stability but not its morphology. Low RNA concentration from stabilized tissue was found. However, there was no crucial issue of protein extraction, degradation or concentration. Tissue morphology was slight different between normal and CL/P tissue

Genomic Alterations, Gene Expression Profiles and Functional Enrichment of Normal-Karyotype Acute Myeloid Leukaemia Based on Targeted Next-Generation Sequencing

Characterising genomic variants is paramount in understanding the pathogenesis and heterogeneity of normal-karyotype acute myeloid leukaemia (AML-NK). In this study, clinically significant genomic biomarkers were ascertained using targeted DNA sequencing and RNA sequencing on eight AML-NK patients’ samples collected at disease presentation and after complete remission. In silico and Sanger sequencing validations were performed to validate variants of interest, and they were followed by the performance of functional and pathway enrichment analyses for overrepresentation analysis of genes with somatic variants. Somatic variants involving 26 genes were identified and classified as follows: 18/42 (42.9%) as pathogenic, 4/42 (9.5%) as likely pathogenic, 4/42 (9.5%) as variants of unknown significance, 7/42 (16.7%) as likely benign and 9/42 (21.4%) as benign. Nine novel somatic variants were discovered, of which three were likely pathogenic, in the CEBPA gene with significant association with its upregulation. Transcription misregulation in cancer tops the affected pathways involving upstream genes (CEBPA and RUNX1) that were deregulated in most patients during disease presentation and were closely related to the most enriched molecular function gene ontology category, DNA-binding transcription activator activity RNA polymerase II-specific (GO:0001228). In summary, this study elucidated putative variants and their gene expression profiles along with functional and pathway enrichment in AML-NK patients.This research was funded by Universiti Sains Malaysia, grant number GIPS-PhD 311/PPSP/4404814 (23 April 2020). Universiti Sains Malaysia funded the APC

Potential association of nicotinamide on the telomerase activity and telomere length mediated by PARP-1 mechanism in myeloid cancer

Administration of nicotinamide is affecting various types of cells through its survival, maturation, and differentiation. Nicotinamide as part of vitamin B3, plays an important role in DNA repair and maintenance of the genomic stability which related to its function as a NAD+ precursor that involve in many biological processes. During DNA breaks, PARP-1 mechanism will be activated and use NAD+ as a substrate in process of DNA damage and repair that will result to either cell repair and cell death. In the meantime, in presence of nicotinamide that is also acting as a PARP-1 inhibitor, causing inability of the repair mechanism to fix the entire DNA damage which also lead to the cell death. Therefore, loss of PARP-1 enzyme will cause disturbance in the DNA repair process. Telomere shortening rate was reduced in the presence of nicotinamide that might related with telomerase enzyme which able to maintain the telomere length of the cell. Other than that, telomere also can be influenced by PARP-1 activity where it might show some correlation between nicotinamide, telomere and telomerase that could related with PARP-1 mechanism. Currently, there is no treatment options that respond effectively in chronic myeloid leukemia (CML) in blast crisis (BC) phase without any side effect and it is require an identification of new drug therapies to treat the CML patients. By understanding the role and potential of nicotinamide relation with PARP-1 mechanism in telomere and telomerase status may improve the therapeutic strategy for chronic myeloid leukemia

AST, ALT, Bilirubin and AST/ALT Ratio role; Covid- 19 Patients

Background Impaired liver function upon admission has been linked to the severity of COVID-19 infection, yet the data is debated [1]. Therefore, this retrospective study aimed to evaluate the liver function among COVID-19 patients during hospitalization and its association with the disease severity. Methodology The patient aged 18 to 80 with positive COVID-19 at Hospital Raja Perempuan Zainab II (HRPZ II), Kota Bharu, Kelantan, with available AST, ALT, Bilirubin, and AST/ALT ratio data on admission, were retrospectively evaluated from March 2021 to March 2022. Disease severity was categorized based on Annex 2e guidelines by Malaysia's Ministry of Health, which further classified them into mild to moderate disease (Stage 1-3) and severe to critical illness (Stage 4-5). The AST, ALT, Bilirubin, and AST/ALT ratio levels on Day 1 admission were archived from the electronic medical record system and compared between the two groups. The statistical analysis was using SPSS version 27. This study was approved by (JEPeM-USM) protocol code USM/JEPeM/21100691 and Ministry of Health Malaysia NMRR-21-762-58458 (IIR). Results and Discussion The study included a total of 168 COVID-19 patients with a mean (SD) age of 46.67(16.10) for mild to moderate and 56.66(12.41) for severe to critical. There is a significant age group for both groups (p-value=0.002). During hospitalization, 16(14.41%) patients progressed to death from severe to critically ill patients. Upon admission, the median (IQR) of AST and ALT were significantly higher in the severe to critical group compared to in the mild to moderate group, [AST; 39.0(49.0) and 24.0(14.0), ALT 38.0(43.0) and 21.0(18.0)], p<0.05. However, no significant difference between both groups for bilirubin level and AST/ALT ratio. Non-survivors had a higher AST and ALT level compared to survivors, with a median (IQR) of [AST 98.0(88.0) and 32.0 (26.0), ALT of 67.5(90.0) and 28.0(31.0), (p<0.05). Similarly, no significant difference between non-survivors and survivors for bilirubin and AST/ALT ratio. Our study support that, abnormal liver function at admission has been shown to be associated with the disease severity and mortality of COVID-19 infection. However, there is also a need to observe the COVID-19 survivors' hepatobiliary sequelae and dynamic liver function changes following hospital discharge. Conclusion Abnormal AST and ALT level at admission has been shown to be associated with the disease severity and mortality of COVID-19 infection. Further study needed to evaluate liver damage in COVID-19 post-discharge

Determination of allelic imbalance in childhood acute lymphoblastic leukaemia

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Genetic Aberrations in Childhood Acute Lymphoblastic Leukaemia: Application of High-Density Single Nucleotide Polymorphism Array

Screening of the entire human genome using high-density single nucleotide polymorphism array (SNPA) has become a powerful technique used in cancer genetics and population genetics studies. The GeneChip® Mapping Array, introduced by Affymetrix, is one SNPA platform utilised for genotyping studies. This GeneChip system allows researchers to gain a comprehensive view of cancer biology on a single platform for the quantification of chromosomal amplifications, deletions, and loss of heterozygosity or for allelic imbalance studies. Importantly, this array analysis has the potential to reveal novel genetic findings involved in the multistep development of cancer. Given the importance of genetic factors in leukaemogenesis and the usefulness of screening the whole genome, SNPA analysis has been utilised in many studies to characterise genetic aberrations in childhood acute lymphoblastic leukaemia

Molecular genetic analysis of phosphatase and tensin homolog and p16 tumor suppressor genes in patients with malignant glioma

Object. Several genes have been shown to carry mutations in human malignant gliomas, including the phosphatase and tensin homolog (PTEN) deleted on chromosome 10 and p16 tumor suppressor genes. Alterations of this gene located on chromosome 10 q23 and 9p21, respectively, may contribute to gliomagenesis. In this study, the authors analyzed 20 cases of malignant gliomas obtained in patients living on the east coast of Malaysia to investigate the possibilities of involvement of the PTEN and p16 genes. Methods. Samples of DNA were amplified by polymerase chain reaction (PCR), analyzed by single-stranded conformation polymorphism (SSCP), and subsequently by sequencing. Two cases of glioblastoma multiforme, three cases of anaplastic astrocytoma, one case of anaplastic pleomorphic xanthoastrocytoma, and one case of anaplastic ependymoma showed SSCP band shifts in PTEN mutational analyses. The DNA sequencing analyses of these samples revealed missense and nonsense mutations, with cluster of mutations in the region 5� to the core phosphatase motif of exon 5 and the 5�-end of exon 6. No abnormal migration shifts were detected in the glioma samples analyzed for point mutations of the p16 gene. Homozygous deletions of p16 were also not detected in all samples. Conclusions. These findings indicate that mutations of the PTEN genes were likely to contribute to the tumorigenesis and morphological transformations of gliomas. In addition, the alterations of the p16 gene might not play a major role in tumorigenesis of malignant gliomas in Malaysian patients

Genetic Profiles and Risk Stratification in Adult De Novo Acute Myeloid Leukaemia in Relation to Age, Gender, and Ethnicity: A Study from Malaysia

Hitherto, no data describing the heterogeneity of genetic profiles and risk stratifications of adult acute myeloid leukaemia (AML) in Southeast Asia are reported. This study assessed genetic profiles, Moorman’s hierarchical classification, and ELN 2017-based risk stratifications in relation to age, gender, and ethnicity in Malaysian adult AML patients. A total of 854 AML patients: male (52%), female (48%) were recruited comprising three main ethnic groups: Malays (59%), Chinese (32%) and Indians (8%). Of 307 patients with abnormal karyotypes: 36% exhibited translocations; 10% deletions and 5% trisomies. The commonest genotype was FLT3-ITD-NPM1wt (276/414; 66.7%). ELN 2017 risk stratification was performed on 494 patients, and 41% were classified as favourable, 39% as intermediate and 20% as adverse groups. More females (47%) were in the favourable risk group compared to males (37%), whereas adverse risk was higher in patients above 60 (24%) of age compared to below 60 (18%) patients. We observed heterogeneity in the distribution of genetic profiles and risk stratifications between the age groups and gender, but not among the ethnic groups. Our study elucidated the diversity of adult AML genetic profiles between Southeast Asians and other regions worldwide