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Fulltext Thymoquinone-loaded nanostructured lipid carrier reduces proliferation of human liver cancer cells, HepG2

Nurfahima Mustafa Azmy, Aminah Suhaila Haron, Sharifah Sakinah Syed Alwi

Malaysian Journal of Medicine and Health Sciences, 2019;15(202):38-43.
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Introduction: Hepatocellular carcinoma is one of the most common cancers that affected human in more than half of the world population. Although there is yet any alternatives treatment found for this disease, the antitumor property of thymoquinone has been well studied in most of cancer cell lines. Nonetheless, poor bioavailability of TQ limits its efficiency. The encapsulation form of TQ, TQ-NLC is suggested to enhance its bioavailability as well as cytotoxicity towards cancer cells via increasing resistance time and targeting drug to specified location in the body. Therefore, it is a great advantage to look at the effects of TQ-NLC towards HepG2. This study is design to look at the anti-pro- liferative effect of TQ-NLC on HepG2 and the changes in the cells morphology. Methods: Both cells were bought from ATCC and cultured in supplemented DMEM. Cell viability was determined via MTT assay. Pro-apoptotic effect of TQ-NLC was further confirmed with Annexin V staining. Morphology hallmarks of apoptosis of treated cells were also analysed using inverted microscope. Images were captured at 24, 48 and 72 hours. Results: TQ-NLC was very potent towards HepG2 compared to 3T3 with the relative IC50 of 25 µM. TQ-NLC was also more potent compared to the non-encapsulated form, TQ. Further analysis confirmed that TQ-NLC capable to increase the percentage of apoptotic cells in time-dependent manner. Qualitatively, all treated cells displayed the apoptosis morphology with increasing concentration and longer time-point. Conclusion: TQ-NLC showed greater cytotoxic effects towards HepG2 which was further confirmed with the morphological analysis.
Fulltext Analysis of air humidity and temperature for post lightning circumstances

Sharifah Sakinah, Tuan Othman, Jasronita, Jasni, Mohd Nazim, Mohtar

Pertanika Journal of Science & Technology, 2017;25(102):79-86.
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Lightning is a natural phenomenon that generates a high electric field during thunderstorm. It has been
reported that lightning strikes amid storms can occur around 100 times per second. The atmospheric
electric field is an imperative parameter during a thunderstorm. Therefore, monitoring the electric field
and its parameters is the best way for local lightning forecast. The electric field monitoring data can
validate the accuracy of weather prediction in a local area from meteorology department or by using
equipment specially designed to measure this electric field that exists when the phenomenon of lightning occurs. In this paper, the relationship between lightning, air humidity and temperature is discussed to understand the post lightning effect on these electric parameters. Additionally, the characteristics of the parameters are observed and analysed.
Fulltext Cytotoxic effect of 2,6-bis(4-Hydroxy-3-Methoxybenzylidene) cyclohexanone (BHMC) and curcumin on human liver cancer cells, HepG2

Sharifah Sakinah Syed Alwi, Syazwan Zahari, Aminah Suhaila Haron, Henna Roshini Alexander

Malaysian Journal of Medicine and Health Sciences, 2019;15(202):44-50.
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Introduction: Curcumin is an active constituent derived from turmeric with a variety of pharmacological activities. It suppressed cell proliferation and induced apoptosis in several cancer cell lines. However, due to its poor bio- availability, derivative analogue of curcumin has been synthesized to enhance the drug-like effects. BHMC was synthesized by removing β-diketone moiety from curcumin structure and modify it into conjugated double bonds. It has been proved to exhibit stronger anticancer effects with improved bioavailability compared to curcumin. Ob- jective: This study aims to investigate the toxicity effect of BHMC and curcumin on human liver cancer, HepG2 and non-cancer mouse fibroblast, 3T3. Methods: Both cell lines were purchased from ATCC and cultured in sup- plemented DMEM. Cell viability was determined via MTT assay and confirmed with trypan blue assay. Morphology hallmarks of apoptosis of both treated cells were analyzed using inverted microscope at 40X magnifications. Results: BHMC and curcumin were very potent towards HepG2 and normal 3T3. These data were further confirmed with trypan blue assay which showed that both compounds significantly reduced the percentage of HepG2 and 3T3 cells viability. Both treated cells also displayed all the morphology hallmarks of apoptosis upon treatment. Conclusion: BHMC has a greater cytotoxicity effect on HepG2 compared to curcumin despite its non-selective cytotoxicity effect on non-cancer 3T3.
Fulltext Dynamic similarity distance with mean average precision tool

Nur Atikah Arbain, Mohd Sanusi Azmi, Sharifah Sakinah Syed Ahmad, Azah Kamilah Muda, Intan Ermahami A. Jalil, King Ming Tiang

Pertanika Journal of Science & Technology, 2017;25(106):11-18.
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In recent years, many classification models have been developed and applied to increase their accuracy. The concept of distance between two samples or two variables is a fundamental concept in multivariate analysis. This paper proposed a tool that used different similarity distance approaches with ranking method based on Mean Average Precision (MAP). In this study, several similarity distance methods were used, such as Euclidean, Manhattan, Chebyshev, Sorenson and Cosine. The most suitable distance measure was based on the smallest value of distance between the samples. However, the real solution showed that the results were not accurate as and thus, MAP was considered the best approach to overcome current limitations.
Fulltext LEP G2548A polymorphism is associated with increased serum leptin and insulin resistance among T2DM Malaysian patients

Ali LA, Jemon K, Latif NA, Bakar SA, Alwi SSS

Biomedicine (Taipei), 2022;12(3):1-11.
PMID: 36381191 DOI: 10.37796/2211-8039.1326

Background: Type 2 diabetes mellitus (T2DM) is a chronic metabolic syndrome that is rapidly increasing across the world, especially in Malaysia. Leptin plays a vital role in the regulation of metabolism through its effect on peripheral tissues. G2548A polymorphism in the LEP gene promoter has been associated with insulin resistance, leptin, and type 2 diabetes mellitus across different population, but has not been inclusively reported within the Malaysian population.
Objective: Thus, our study aimed to investigate the impact of G2548A polymorphism on serum leptin levels and insulin resistance among Malaysian T2DM patients.
Methods: This case-control study involved 150 T2DM patients and 150 non-diabetic volunteers from ethnic Malays, Chinese and Indians. Genotyping of G2548A polymorphism was carried out using PCR-RFLP. Serum leptin and insulin levels were determined via ELISA. ANOVA and Chi-square tests were used to determine the distribution of genotypes and allelic frequencies based on serum leptin and insulin levels.
Results: Frequency of AA genotype and A allele of G2548A variant were significantly (P < 0.05) higher in T2DM patients of Malay and Indian ethnicities (4%, 35%, and 36%, 57%, respectively) as compared to the control groups (0%, 22%, and 18%, 35%, respectively). Fasting serum leptin levels were significantly (P < 0.001) higher in T2DM patients compared to non-diabetic subjects (166.78 pg/ml, 101.94 pg/ml, respectively). Additionally, elevated serum leptin, insulin levels, and BMI in diabetic patients were found to be associated with the AA genotype of this variant, compared to GG, and GA genotypes (P < 0.05).
Conclusion: Our findings suggest a significant association between G2548A polymorphism among Malaysian T2DM subjects, particularly among Malay and Indian ethnic groups. Moreover, the A allele frequency of the G2548A variant significantly increased the risk of T2DM and is significantly associated with increased serum leptin, insulin levels, and elevated BMI.
Fulltext Insomnia and depression levels among Malaysian undergraduate students in the Faculty of Medicine and Health Sciences (FMHS), Universiti Putra Malaysia (UPM) during Movement Control Order (MCO)

Iqbal RM, Binti Riza Effendi NI, Syed Alwi SS, Saidi HI, Sarchio SNE

PLoS One, 2023;18(10):e0283098.
PMID: 37816038 DOI: 10.1371/journal.pone.0283098

Rapid outbreak of coronavirus disease 2019 has caused the implementation of the movement control order (MCO) which aimed to reduce the spread in Covid-19 infections. While some may find it easy to adjust to the new norm, others found it difficult to switch from their normal routines and habits as according to the MCO SOP. This resulted in a more frequent insomnia and depression that subsequently impacted their mental health. Insomnia and depression levels are examined in this study as they relate to the Covid-19 Pandemic and the MCO among Malaysian undergraduate health sciences students at the Faculty of Medicine and Health Sciences, UPM. Random sampling methods were utilised with consideration of inclusion and exclusion criteria. The Patient Health Questionnaire-9 (PHQ-9) and the Insomnia Severity Index (ISI) were the instrument packages used in this investigation. An internet platform was used to distribute the questionnaire. Based on the results, it is concluded that depression and insomnia are significantly correlated, with a p-value of 0.05. This study also revealed the link between the severity of insomnia and the severity of depression among UPM students studying health sciences. The percentage of students with depression and insomnia was rather high (54.9% and 33.9%, respectively), and this occurred during the second wave of Covid-19 cases in Malaysia.
Fulltext Migration and Proliferation Effects of Thymoquinone-Loaded Nanostructured Lipid Carrier (TQ-NLC) and Thymoquinone (TQ) on In Vitro Wound Healing Models

Alexander HR, Syed Alwi SS, Yazan LS, Zakarial Ansar FH, Ong YS

Evid Based Complement Alternat Med, 2019;2019:9725738.
PMID: 31915456 DOI: 10.1155/2019/9725738

Wound healing is a regulated biological event that involves several processes including infiltrating leukocyte subtypes and resident cells. Impaired wound healing is one of the major problems in diabetic patients due to the abnormal physiological changes of tissues and cells in major processes. Thymoquinone, a bioactive compound found in Nigella sativa has been demonstrated to possess antidiabetic, anti-inflammatory, and antioxidant effects. Today, the rapidly progressing nanotechnology sets a new alternative carrier to enhance and favour the speed of healing process. In order to overcome its low bioavailability, TQ is loaded into a colloidal drug carrier known as a nanostructured lipid carrier (NLC). This study aimed to determine the effect of TQ-NLC and TQ on cell proliferation and migration, mode of cell death, and the antioxidant levels in normal and diabetic cell models, 3T3 and 3T3-L1. Cytotoxicity of TQ-NLC and TQ was determined by MTT assay. The IC10 values obtained for 3T3-L1 treated with TQ-NLC and TQ for 24 hours were 4.7 ± 3.3 and 5.3 ± 0.6 μM, respectively. As for 3T3, the IC10 values obtained for TQ-NLC and TQ at 24 hours were 4.3 ± 0.17 and 3.9 ± 2.05 μM, respectively. TQ-NLC was observed to increase the number of 3T3 and 3T3-L1 healthy cells (87-95%) and gradually decrease early apoptotic cells in time- and dose-dependant manner compared with TQ. In the proliferation and migration assay, 3T3-L1 treated with TQ-NLC showed higher proliferation and migration rate (p < 0.05) compared with TQ. TQ-NLC also acted as an antioxidant by reducing the ROS levels in both cells after injury at concentration as low as 3 μM. Thus, this study demonstrated that TQ-NLC has better proliferation and migration as well as antioxidant effect compared with TQ especially on 3T3-L1 which confirms its ability as a good antidiabetic and antioxidant agent.
Fulltext Anti-breast cancer synthetic peptides derived from the Anabas testudineus skin mucus fractions

Najm AAK, Azfaralariff A, Dyari HRE, Othman BA, Shahid M, Khalili N, et al.

Sci Rep, 2021 11 30;11(1):23182.
PMID: 34848729 DOI: 10.1038/s41598-021-02007-6

Previous study has shown the antimicrobial activities of mucus protein extracted from Anabas testudineus. In this study, we are interested in characterizing the anticancer activity of the A. testudineus antimicrobial peptides (AMPs). The mucus was extracted, fractioned, and subjected to antibacterial activity testing to confirm the fish's AMPs production. The cytotoxic activity of each fraction was also identified. Fraction 2 (F2), which shows toxicity against MCF7 and MDA-MB-231 were sent for peptide sequencing to identify the bioactive peptide. The two peptides were then synthetically produced and subjected to cytotoxic assay to prove their efficacy against cancer cell lines. The IC50 for AtMP1 against MCF7 and MDA-MB-231 were 8.25 ± 0.14 μg/ml and 9.35 ± 0.25 μg/ml respectively, while for AtMP2 it is 5.89 ± 0.14 μg/ml and 6.97 ± 0.24 μg/ml respectively. AtMP1 and AtMP2 treatment for 48 h induced breast cancer cell cycle arrest and apoptosis by upregulating the p53, which lead to upregulate pro-apoptotic BAX gene and downregulate the anti-apoptotic BCL-2 gene, consequently, trigger the activation of the caspase-3. This interaction was supported by docking analysis (QuickDBD, HPEPDOCK, and ZDOCK) and immunoprecipitation. This study provided new prospects in the development of highly effective and selective cancer therapeutics based on antimicrobial peptides.
Fulltext Cytotoxic Effect of Thymoquinone-Loaded Nanostructured Lipid Carrier (TQ-NLC) on Liver Cancer Cell Integrated with Hepatitis B Genome, Hep3B

Haron AS, Syed Alwi SS, Saiful Yazan L, Abd Razak R, Ong YS, Zakarial Ansar FH, et al.

Evid Based Complement Alternat Med, 2018;2018:1549805.
PMID: 30186351 DOI: 10.1155/2018/1549805

Thymoquinone (TQ), a bioactive compound found in Nigella sativa, cannot be orally consumed due to its lipophilicity. In order to overcome this low bioavailability, TQ is loaded into a colloidal drug carrier known as a nanostructured lipid carrier (NLC). This study aims to determine the antiproliferative effects of TQ and TQ-NLC on liver cancer cells integrated with the hepatitis B genome, Hep3B. The Hep3B was treated with TQ or TQ-NLC for 24, 48, and 72 hours via MTT assay. The results confirm that TQ or TQ-NLC inhibited the growth of Hep3B at IC50 <16.7 μM for 72 hours. TQ was also found to induce cell cycle arrest at the G1 checkpoint while TQ-NLC induced non-phase-specific cell cycle arrest. Further analysis using Annexin V staining confirmed the apoptotic induction of TQ or TQ-NLC via activation of caspases-3/7. In ROS management, TQ acted as a prooxidant (increased the level of ROS), while TQ-NLC acted as an antioxidant (reduced the level of ROS). Molecular analysis demonstrated that the GSH system and the Nrf2/Keap1 signaling pathway in Hep3B influenced the differential responses of the cells towards TQ or TQ-NLC. Hence, this study demonstrated that TQ and TQ-NLC have in vitro anticancer effects on the Hep3B.
Fulltext Chemopreventive Properties and Toxicity of Kelulut Honey in Sprague Dawley Rats Induced with Azoxymethane

Saiful Yazan L, Muhamad Zali MF, Mohd Ali R, Zainal NA, Esa N, Sapuan S, et al.

Biomed Res Int, 2016;2016:4036926.
PMID: 27525267 DOI: 10.1155/2016/4036926

Ethnopharmacological Relevance. Colon cancer has been a major problem worldwide. Kelulut honey (KH) is produced by the stingless bees from Trigona species and has strong antioxidant activities that could be one of the potential chemopreventive agents from natural resources. Aim of This Study. This study investigated the chemopreventive properties and toxicity of KH in Sprague Dawley rats induced with azoxymethane (AOM). Material and Method. Twenty-four male Sprague Dawley rats aged 5 weeks were divided into 4 groups: (G1) untreated group not induced with AOM, (G2) untreated group induced with AOM, (G3) treated group induced with AOM, and (G4) treated group not induced with AOM. Injection of AOM (15 mg/kg) was via intraperitoneal route once a week for two subsequent weeks. The treatment groups were given oral administration of KH (1183 mg/kg body weight) twice daily for 8 weeks. Results. Treatment with KH significantly reduced the total number of aberrant crypt foci (ACF) and aberrant crypts (AC) and crypt multiplicity. KH was not toxic to the animals since the level of blood profile parameters, liver enzymes, and kidney functions was in normal range. Conclusions. The current finding shows that KH has chemopreventive properties in rats induced with colorectal cancer and also was found not toxic towards the animals.
Fulltext Vitamins D and E Stimulate the PI3K-AKT Signalling Pathway in Insulin-Resistant SK-N-SH Neuronal Cells

Zaulkffali AS, Md Razip NN, Syed Alwi SS, Abd Jalil A, Abd Mutalib MS, Gopalsamy B, et al.

Nutrients, 2019 Oct 19;11(10).
PMID: 31635074 DOI: 10.3390/nu11102525

This study investigated the effects of vitamins D and E on an insulin-resistant model and hypothesized that this treatment would reverse the effects of Alzheimer's disease (AD) and improves insulin signalling. An insulin-resistant model was induced in SK-N-SH neuronal cells with a treatment of 250 nM insulin and re-challenged with 100 nM at two different incubation time (16 h and 24 h). The effects of vitamin D (10 and 20 ng/mL), vitamin E in the form of tocotrienol-rich fraction (TRF) (200 ng/mL) and the combination of vitamins D and E on insulin signalling markers (IR, PI3K, GLUT3, GLUT4, and p-AKT), glucose uptake and AD markers (GSK3β and TAU) were determined using quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). The results demonstrated an improvement of the insulin signalling pathway upon treatment with vitamin D alone, with significant increases in IR, PI3K, GLUT3, GLUT4 expression levels, as well as AKT phosphorylation and glucose uptake, while GSK3β and TAU expression levels was decreased significantly. On the contrary, vitamin E alone, increased p-AKT, reduced the ROS as well as GSK3β and TAU but had no effect on the insulin signalling expression levels. The combination of vitamins D and E only showed significant increase in GLUT4, p-AKT, reduced ROS as well as GSK3β and TAU. Thus, the universal role of vitamin D, E alone and in combinations could be the potential nutritional agents in restoring the sensitivity of neuronal cells towards insulin and delaying the pathophysiological progression of AD.
Fulltext Antiulcer properties of Kelulut honey against Ethanol-induced gastric ulcer

Latifah Saiful Yazan, Nurul Amira Zainal, Muhamad Firdaus Shyfiq Muhamad Zali, Gopalsamy, Banulata, Ling, Voon Fui, Aminah Suhaila Haron, et al.

Pertanika Journal of Science & Technology, 2018;26(1):121-132.
MyJurnal

Ulcers in the gastrointestinal tract refer to any appreciable depth of break in the mucosa lining that may involve submucosa. Common types of ulcer include peptic, gastric and duodenal ulcer, which may lead to chronic inflammation. Ulcers may be caused by excessive alcohol intake or prolonged use of non-steroidal anti-inflammatory drugs (NSAID), in addition to several other factors. Conventional medication such as Omeprazole (proton pump inhibitor) and Ranitidine (H2 blockers) for management of ulcers may cause severe side effects such as myelosupression and abnormal heart rhythm. This has driven researchers to explore the potential of natural products for management of ulcers with reduced side effects. Kelulut honey (KH) is a type of honey that is produced by stingless bees from the Trigona species. It is believed to have a lot of medicinal properties such as being antimicrobial, antioxidant and antidiabetic. Yet, no scientific study has been carried out on its antiulcer properties. This study was carried out to determine the antiulcer properties of KH. Eighteen male Sprague dawley rats (5 to 6 weeks old, weighing between 200 and 300 g) were divided into three groups (n=6). The groups were 1) normal control group (without ulcer, without KH), 2) positive control group (with ulcer, without KH) and 3) treatment group (with ulcer, treated with KH). The treatment, KH (1183 mg/kg), was given twice daily for 30 consecutive days by oral administration. On Day 31, the rats were induced with absolute ethanol (5 mL/kg) via oral administration after being fasted for 24 h and were sacrificed 15 min after the induction. The stomach was collected for macroscopic and histopathological evaluation. Pretreatment with KH significantly reduced (p

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