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Cytotoxicity evaluation of ketoprofen found in pharmaceutical wastewater on HEK 293 cell growth and metabolism

Zulkarnain NN, Anuar N, Johari NA, Sheikh Abdullah SR, Othman AR

Environ Toxicol Pharmacol, 2020 Nov;80:103498.
PMID: 32950717 DOI: 10.1016/j.etap.2020.103498

Inefficient ketoprofen removal from pharmaceutical wastewater may negatively impact the ecosystem and cause detrimental risks to human health. This study was conducted to determine the cytotoxicity effects of ketoprofen on HEK 293 cell growth and metabolism, including cyclooxygenase-1 (COX-1) expression, at environmentally relevant concentrations. The cytotoxic effects were evaluated through the trypan blue test, DNS assay, MTT assay, and the expression ratio of the COX-1 gene. The results of this study show insignificant (p > 0.05) cytotoxic effects of ketoprofen on cell viability and cell metabolism. However, high glucose consumption rates among the treated cells cause an imitation of the Warburg effect, which is likely linked to the development of cancer cells. Apart from that, the upregulation of COX-1 expression among the treated cells indicates remote possibility of inflammation. Although no significant cytotoxic effects of ketoprofen were detected throughout this study, the effects of prolonged exposure of residual ketoprofen need to be evaluated in the future.
Fulltext Reconstruction of Secondary Type Congenital Hallux Varus with Modification of the Farmer Technique for Cover of Skin Defect: Report of Three Cases

Sulaiman AR, Munajat I, M-Yusuf H, Nik-Jaffar N, Zarullail NH, Mohd EF, et al.

Malays Orthop J, 2019 Nov;13(3):72-76.
PMID: 31890115 DOI: 10.5704/MOJ.1911.013

The reconstruction of hallux varus deformity involves the release of contracted medial structure and realignment of the phalange, leaving a significant skin defect which requires cover. Farmer described proximal based rotational skin flap from the first web space to cover the defect. This technique may compromise the circulation to the flap and risk to the lateral digital vessels. We modified his technique to address these issues. We report a successful reconstruction using the Farmer's technique on one patient and a modified technique on three patients. We used the excess skin from the extra digit to cover the medial defect. We found this modified technique of skin cover safe without risk of injuring the neurovascular bundle. There was no recurrence of deformity at last foolow-up. All patients were able to wear normal shoes.
Fulltext Optimization of nucleic acid extraction and amplification of a thiamine biosynthesis gene fragment from selected Malaysian seaweeds

Md Ahaik FA, Mohd Taufik SH, Faiqah Johari NA, Zainal Abidin AA, Balia Yusof ZN

Genes Genet Syst, 2023 Feb 22;97(5):247-256.
PMID: 36631109 DOI: 10.1266/ggs.22-00088

Obtaining high-quality nucleic acid extracted from seaweeds is notoriously difficult due to contamination with polysaccharides and polyphenolic compounds after cell disruption. Specific methods need to be employed for RNA isolation in different seaweed species, and therefore studies of the thiamine biosynthesis pathway have been limited. Two selected Malaysian species which are highly abundant and underutilized, namely Gracilaria sp. and Padina sp., representing the red and brown seaweeds, respectively, were collected to develop optimized total RNA extraction methods. Prior to that, DNA was extracted, and amplification of the 18S rRNA gene and the THIC gene (encoding the first enzyme in the pyrimidine branch of the thiamine biosynthesis pathway) from the DNA template was successful in Gracilaria sp. only. RNA was then extracted from both seaweeds using three different existing methods, with some modifications, using cetyltrimethylammonium bromide, guanidine thiocyanate and sodium dodecyl sulphate. Methods I and III proved to be efficient for Padina sp. and Gracilaria sp., respectively, for the extraction of highly purified RNA, with A260/A280 values of 2.0 and 1.8. However, amplification of the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase and the THIC gene was successful in only Gracilaria sp. cDNA derived from extracted RNA. Further modifications are required to improve the exploitation of nucleic acids from brown seaweeds, which has been proven to be difficult. This work should pave the way for molecular studies of seaweeds generally and for the elucidation, specifically, of the thiamine biosynthesis pathway.
Fulltext Sylvatic dengue virus type 4 in Aedes aegypti and Aedes albopictus mosquitoes in an urban setting in Peninsular Malaysia

Johari NA, Voon K, Toh SY, Sulaiman LH, Yap IKS, Lim PKC

PLoS Negl Trop Dis, 2019 Nov;13(11):e0007889.
PMID: 31730672 DOI: 10.1371/journal.pntd.0007889

Dengue fever is endemic in Malaysia, contributing to significant economic and health burden in the country. Aedes aegypti and Ae. albopictus are the main vectors of the dengue virus (DENV), which circulates in sylvatic and human transmission cycles and has been present in Malaysia for decades. The study investigated the presence and distribution of DENV in urban localities in the Klang Valley, Peninsular Malaysia. A total of 364 Ae. aegypti and 1,025 Ae. albopictus larvae, and 10 Ae. aegypti and 42 Ae. albopictus adult mosquitoes were screened for the presence of DENV. In total, 31 (2.2%) samples were positive, of which 2 Ae. albopictus larvae were co-infected with two serotypes, one with DENV-2 and DENV-3 and the other with DENV-3 and DENV-4. Phylogenetic analysis determined that the isolates belonged to DENV-1 genotype I (1 Ae. aegypti adult), DENV-2 (1 Ae. albopictus larva), DENV-3 genotype V (3 Ae. aegypti larvae and 10 Ae. albopictus larvae) and DENV-4 genotype IV (6 Ae. aegypti larvae and 12 Ae. albopictus larvae), a sylvatic strain of DENV-4 which was most closely related with sylvatic strains isolated from arboreal mosquitoes and sentinel monkeys in Peninsular Malaysia in the 1970s. All four DENV serotypes were co-circulating throughout the study period. The detection of a sylvatic strain of DENV-4 in Ae. aegypti and Ae. albopictus mosquitoes in urban areas in Peninsular Malaysia highlights the susceptibility of these vectors to infection with sylvatic DENV. The infectivity and vector competence of these urban mosquitoes to this strain of the virus needs further investigation, as well as the possibility of the emergence of sylvatic virus into the human transmission cycle.
Physicochemical homogeneity of stingless bee honey (Heterotrigona itama) produced in the west coast, east coast and inland area of Peninsular Malaysia

Ramlan NAFM, Mohamad Azman E, Muhammad K, Jusoh AZ, Johari NA, Yusof YA, et al.

J Sci Food Agric, 2024 Feb;104(3):1756-1767.
PMID: 37862235 DOI: 10.1002/jsfa.13067

BACKGROUND: The nutritional composition of stingless bee honey (SBH) can be affected by different climates and soil composition across different geographical areas. However, the range of attributes set for a honey quality standard should be inclusive. This study analysed the sugar profile's physiochemical properties, including quantifying the rare sugar trehalulose, organic acid and mineral composition of SBH collected from inland, and west and east coasts of Peninsular Malaysia. Forty-three SBH (Heterotrigona itama) samples were collected and labelled as <20 and <40 West Coast (<20WC, <40WC), <20 and <40 East Coast (<20EC, <40EC) and Inland, according to their distance from the coasts.
RESULTS: The moisture, pH and sugar composition of all SBH samples adhered to the Malaysian Kelulut Honey Standard (MS2683:2017) but not to the International Codex Standard (CODEX) for honey. Trehalulose presence in all samples, regardless of geographical area, was predominant alongside fructose and glucose. Only hydroxymethylfurfural (HMF) content and electrical conductivity (EC) results complied with both standards. The principal component analysis biplot showed that the discrimination of SBH according to the five different areas was not feasible, indicating sample homogeneity.
CONCLUSION: The physicochemical evaluation of SBH from Peninsular Malaysia shows mainly homogeneous attributes of samples across geographical locations. These findings demonstrated that the current MS2683:2017 is relevant and accommodates all SBH of H. itama species produced in Peninsular Malaysia. Furthermore, the trehalulose range calculated in this study can be implemented as a new benchmark for the indicator of SBH honey quality standard by national and international food standard committees. © 2023 Society of Chemical Industry.
Fulltext A novel strategy for community screening of SARS-CoV-2 (COVID-19): Sample pooling method

Lim KL, Johari NA, Wong ST, Khaw LT, Tan BK, Chan KK, et al.

PLoS One, 2020;15(8):e0238417.
PMID: 32857823 DOI: 10.1371/journal.pone.0238417

The rapid global spread of the coronavirus disease (COVID-19) has inflicted significant health and socioeconomic burden on affected countries. As positive cases continued to rise in Malaysia, public health laboratories experienced an overwhelming demand for COVID-19 screening. The confirmation of positive cases of COVID-19 has solely been based on the detection of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) using real-time reverse transcription polymerase chain reaction (qRT-PCR). In efforts to increase the cost-effectiveness and efficiency of COVID-19 screening, we evaluated the feasibility of pooling clinical Nasopharyngeal/Oropharyngeal (NP/OP) swab specimens during nucleic acid extraction without a reduction in sensitivity of qRT-PCR. Pools of 10 specimens were extracted and subsequently tested by qRT-PCR according to the WHO-Charité protocol. We demonstrated that the sample pooling method showed no loss of sensitivity. The effectiveness of the pooled testing strategy was evaluated on both retrospective and prospective samples, and the results showed a similar detection sensitivity compared to testing individual sample alone. This study demonstrates the feasibility of using a pooled testing strategy to increase testing capacity and conserve resources, especially when there is a high demand for disease testing.
Fulltext The effects of Piper sarmentosum aqueous extracts on zebrafish (Danio rerio) embryos and caudal fin tissue regeneration

Zainol Abidin IZ, Fazry S, Jamar NH, Ediwar Dyari HR, Zainal Ariffin Z, Johari AN, et al.

Sci Rep, 2020 08 25;10(1):14165.
PMID: 32843675 DOI: 10.1038/s41598-020-70962-7

In Malaysia, Piper sarmentosum or 'kaduk' is commonly used in traditional medicines. However, its biological effects including in vivo embryonic toxicity and tissue regenerative properties are relatively unknown. The purpose of this study was to determine zebrafish (Danio rerio) embryo toxicities and caudal fin tissue regeneration in the presence of P. sarmentosum aqueous extracts. The phytochemical components and antioxidant activity of the extract were studied using GC-MS analysis and DPPH assay, respectively. Embryo toxicity tests involving survival, heartbeat, and morphological analyses were conducted to determine P. sarmentosum extract toxicity (0-60 µg/mL); concentrations of 0-400 µg/mL of the extract were used to study tissue regeneration in the zebrafish caudal fin. The extract contained several phytochemicals with antioxidant activity and exhibited DPPH scavenging activity (IC50 = 50.56 mg/mL). Embryo toxicity assays showed that a concentration of 60 μg/mL showed the highest rates of lethality regardless of exposure time. Slower embryogenesis was observed at 40 µg/mL, with non-viable embryos first detected at 50 µg/mL. Extracts showed significant differences (p
Fulltext COVID-19 Sample Pooling: From RNA Extraction to Quantitative Real-time RT-PCR

Voon K, Johari NA, Lim KL, Wong ST, Khaw LT, Wong SF, et al.

Bio Protoc, 2021 May 05;11(9):e4005.
PMID: 34124305 DOI: 10.21769/BioProtoc.4005

The COVID-19 pandemic requires mass screening to identify those infected for isolation and quarantine. Individually screening large populations for the novel pathogen, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), is costly and requires a lot of resources. Sample pooling methods improve the efficiency of mass screening and consume less reagents by increasing the capacity of testing and reducing the number of experiments performed, and are therefore especially suitable for under-developed countries with limited resources. Here, we propose a simple, reliable pooling strategy for COVID-19 testing using clinical nasopharyngeal (NP) and/or oropharyngeal (OP) swabs. The strategy includes the pooling of 10 NP/OP swabs for extraction and subsequent testing via quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR), and may also be applied to the screening of other pathogens.
Fulltext SARS-CoV-2 genomic surveillance in Malaysia: displacement of B.1.617.2 with AY lineages as the dominant Delta variants and the introduction of Omicron during the fourth epidemic wave

Azami NAM, Perera D, Thayan R, AbuBakar S, Sam IC, Salleh MZ, et al.

Int J Infect Dis, 2022 Dec;125:216-226.
PMID: 36336246 DOI: 10.1016/j.ijid.2022.10.044

OBJECTIVES: This study reported SARS-CoV-2 whole genome sequencing results from June 2021 to January 2022 from seven genome sequencing centers in Malaysia as part of the national surveillance program.
METHODS: COVID-19 samples that tested positive by reverse transcription polymerase chain reaction and with cycle threshold values <30 were obtained throughout Malaysia. Sequencing of SARS-CoV-2 complete genomes was performed using Illumina, Oxford Nanopore, or Ion Torrent platforms. A total of 6163 SARS-CoV-2 complete genome sequences were generated over the surveillance period. All sequences were submitted to the Global Initiative on Sharing All Influenza Data database.
RESULTS: From June 2021 to January 2022, Malaysia experienced the fourth wave of COVID-19 dominated by the Delta variant of concern, including the original B.1.617.2 lineage and descendant AY lineages. The B.1.617.2 lineage was identified as the early dominant circulating strain throughout the country but over time, was displaced by AY.59 and AY.79 lineages in Peninsular (west) Malaysia, and the AY.23 lineage in east Malaysia. In December 2021, pilgrims returning from Saudi Arabia facilitated the introduction and spread of the BA.1 lineage (Omicron variant of concern) in the country.
CONCLUSION: The changing trends of circulating SARS-CoV-2 lineages were identified, with differences observed between west and east Malaysia. This initiative highlighted the importance of leveraging research expertise in the country to facilitate pandemic response and preparedness.