This study investigates the antifungal activity of crude ethanolic extracts derived from roselle decoction residues, generated during calyx cordial and juice processing, against Pestalotiopsis sp., a postharvest fungal pathogen associated with fruit rot in wax apples. Valorizing this agro-industrial by-product presents an opportunity to develop sustainable antifungal agents. The experiment evaluated the efficacy of extracts prepared from both fresh and freeze-dried residues, namely roselle calyx (RC), decocted cordial residue (DCR), and decocted juice residue (DJR), at eight different concentrations (10 to 100,000 mg/L) using in vitro and in vivo assays. In vitro results revealed that freeze-dried samples exhibited significantly higher antifungal activity than fresh samples, with complete inhibition observed at lower concentrations (30,000 mg/L) for freeze-dried RC and DCR. DJR showed the lowest efficacy among all calyx sources. In vivo evaluation confirmed that coating wax apples with the most effective extracts reduced disease severity index (DSI) compared to both untreated and chitosan-treated controls. Notably, freeze-dried RC resulted in the lowest DSI, indicating strong protection against fungal infection. The antifungal effects were found to be dose-dependent and involved both fungistatic and fungicidal mechanisms. The superior efficacy of freeze-dried samples is attributed to the better retention of bioactive compounds such as anthocyanins and phenolics, which may disrupt fungal cell integrity and activate plant defense responses. This study supports the use of roselle decoction residues as a low-cost, eco-friendly alternative to synthetic fungicides, aligning with sustainable postharvest disease management strategies.
The advertisement calls of frogs serve as vital communication signals, playing a significant role in species recognition and mate attraction, representing a critical aspect of their behavioural ecology. This study aims to document and analyse the advertisement calls of eight Microhylidae species in various habitats across Sarawak. Using field recordings and acoustic analysis, distinct call characteristics were identified, including frequency, note repetition rate, and pulse structure. Advertisement calls were recorded and analysed using SoundRulerAcousticAnalysis and PraatAcoustic Software. Call characteristics were subsequently compared for species differentiation. The results showed that each species exhibits distinct call characteristics, likely reflecting adaptations to habitat acoustics and conspecific interactions. Among them, Microhyla nepenthicola Das & Haas, 2010 demonstrated the highest rate of note repetition, the greatest number of pulsed notes per call, and the highest dominant frequency, suggesting unique acoustic adaptations within its ecological niche. These findings can contribute to the understanding of species-specific vocalisations and their potential role in mate selection, territorial defence, and environmental adaptation.
Osteosarcoma is a prevalent and aggressive bone malignancy, primarily affecting children and adolescents. While tamoxifen (TAM) is a commonly used chemotherapeutic agent, its application is often limited by adverse side effects. Gallic acid (GA), a naturally occurring polyphenol, has demonstrated promising anticancer properties and may offer a safer alternative or adjunct in cancer therapy. This study investigates the effects of GA alone and in combination with TAM on the viability and morphology of human osteosarcoma (U2OS) cells. An in vitro model was employed using the MTT assay to assess cell viability after 72 hr of treatment. The half maximal inhibitory concentration (IC50) and combination index (CI) were calculated to evaluate cytotoxicity and drug interaction. Three combination ratios of GA:TAM (75:25, 50:50 & 25:75) were tested. GA and TAM alone exhibited IC50 values of 8.258 +/- 1.047 & micro;g/mL and 1.814 +/- 1.205 & micro;g/mL, respectively. The 25:75 combination yielded the lowest IC50 (1.475 +/- 1.189 & micro;g/mL) and a CI of 0.637, indicating a synergistic effect. Other combinations showed higher IC50 and CI values, suggesting reduced efficacy. Morphological analysis using phase contrast microscopy revealed apoptotic features such as cell shrinkage and rounding in treated cells, particularly in the 25:75 combination group. In conclusion, GA, especially when combined with TAM at a 25:75 ratio, enhances therapeutic efficacy while potentially reducing toxicity. This combination may represent a promising strategy for improving osteosarcoma treatment outcomes.
Face masks are a critical component of Personal Protective Equipment (PPE) for occupational safety and health management. Nevertheless, prolonged usage of face masks may expose employees to biological contaminants, such as bacteria, which accumulate on the inner surface of the mask and pose potential health risks. This study involved 216 participants employed by a statutory institution in Malaysia. The participants were assigned to wear three types of face masks: cotton, surgical, and Korean filters for durations of four and eight hr during their respective work shifts, which varied across roles such as office workers, trainers, and cleaners. Bacterial isolates from the masks were analyzed using the MALDI-TOF method for species identification. Twelve bacterial species were successfully identified, including Staphylococcus hominis, Staphylococcus epidermidis, Micrococcus luteus, Bacillus pumilus, Bacillus cereus, Sporosarcina luteola, and Staphylococcus aureus. Most of these species are part of the normal human microbiota but have the potential to act as opportunistic pathogens. Additionally, other bacterial species, particularly spore-forming ones, may have originated from the work environment. To mitigate microbial risks, individuals performing strenuous physical tasks or those with compromised immunity are advised to avoid reusing face masks and to replace them every four hr.
Brachiaria decumbens, commonly known as signal grass, is widely used as livestock feed in tropical regions due to its high nutritional value and availability. However, its high concentration of steroidal saponins poses toxicity risks, raising concerns for small ruminant farmers. Therefore, this study aimed to evaluate the effect of feeding ensiled B. decumbens on the health and performance of sheep under tropical conditions. A total of eighteen six-month-old male Dorper cross sheep were used in a feeding trial and assigned to three treatment groups: the control group (T1), fed Pennisetum purpureum; Treatment 2 (T2), fed fresh B. decumbens; and Treatment 3 (T3), fed ensiled B. decumbens. Over the 98-day experimental period, clinical data were collected weekly. At the end of the study, all 18 sheep were slaughtered for morphometric, gross morphology, and histopathological analyses of their organs. The results revealed significant differences (p<0.05) in the health performance of sheep across feeding groups. Sheep in the T3 group, fed ensiled B. decumbens, exhibited slightly higher body weight gain compared to both the T2 group fed fresh B. decumbens and the control group (T1). Additionally, only T2 sheep exhibited pale ocular mucous membranes during the final 21 days of the study. In terms of organ morphometrics, T1 and T3 sheep had lower organ width, length, and weight compared to T2 sheep. Most vital organs appeared grossly normal across all treatments; however, post-mortem examination of T2 sheep revealed lung lesions, including congestion and pus accumulation in the caudal lobes. Histopathological analysis indicated mild to moderate lesions in various organs of T2 sheep fed fresh B. decumbens, whereas no lesions were observed in the T1 and T3 groups. This study concludes that incorporating ensiled B. decumbens into the diet of small ruminants yields more favorable health and performance outcomes than feeding fresh B. decumbens.
Turmeric (Curcuma longa) is an important herbal plant used in the cosmetic and medical industries. It contains two important products: curcuminoids and volatile oils. Three types of curcuminoids exist: curcumin (the major compound), monodesmethoxycurcumin, and bisdes-methoxycurcumin. Curcuminoids can remedy various diseases such as peptic ulcers, gastric ulcers, dyspepsia, and metabolic and inflammatory conditions. However, monoculture cropping and the practice of rhizome propagation can lead to pathogen contamination, reducing the quality and quantity of curcuminoids. This study aimed to propagate in vitro C. longa plants and improve curcuminoid yield. Rhizomes were propagated using a micropropagation technique, and multiple shoot growth was induced on Murashige and Skoog media supplemented with 5 mgL-1 kinetin and 1 mgL-1 BA (8.00 +/- 0.58 shoots). Shoots grown on MS media with the addition of 5 & micro;M phenylalanine, 5 & micro;M malonic acid, and a growth retardant (ancymidol) exhibited the highest curcuminoid accumulation (2.31 +/- 0.061 & micro;gg-1 dry weight). It can be concluded that the addition of phenylalanine and malonic acid can stimulate curcuminoid biosynthesis. Therefore, supplementing certain precursors, such as leguminous plants and green grasses, as planting material before cultivation can help increase curcuminoid levels.
The utilisation of indigenous ureolytic bacteria for the rehabilitation of soil slope cracks through microbial-induced calcite precipitation (MICP) is cost-effective and eco-friendly. This study was designed to assess the soil's crack-healing potential of the indigenous bacteria, Bacillus cereus. A solution containing the urease-producing B. cereus and B. megaterium (control treatment) and a cementation solution was sprayed onto the cracks; this process was repeated five times. B. cereus treatment induced soil crack repair, with an average reduction in width of 0.20 +/- 0.113 mm from the initial width of 0.45 +/- 0.187 mm, but the decrease in cracks was not significantly different compared to that observed for B. megaterium. Soil treated with B. cereus contained significantly more CaCO3 per 20 g of soil (0.78 +/- 0.102 g vs. 0.59 +/- 0.220 g) compared to B. megaterium and provided optimal growth conditions for the native B. cereus. However, X-ray diffraction analysis revealed that only 1.5% of the precipitated CaCO3 was in the form of calcite, while aragonite constituted 28.5% and vaterite accounted for 70% of the different crystal structures. B. megaterium was composed of 46% calcite, 24.5% aragonite, and 29.5% vaterite crystals. These findings indicate a positive outlook for the use of indigenous microbial-induced calcite precipitation, especially for fracture repair. Further research is required to determine the mechanism underlying the calcification process of native bacteria and the specific conditions necessary to initiate the formation of calcium carbonate biominerals.
Trichomes, which are specialised structures found in the epidermis, are essential for the protection of plants and the secretion of secondary metabolites. This study examines the structures and density of trichomes on the flowers of five selected Hoya Wight. Through the application of light microscopy (LM) and scanning electron microscopy (SEM), we identified the types, morphology and localisation of trichomes in relation to floral glands. The findings indicated the existence of glandular trichomes, simple trichomes and papillae exhibiting various shapes, including conical, filamentous, finger-like, falcate and stalked capitate apical prostrusion located on the adaxial epidermis of corolla petals and pedicels. This investigation deepens the comprehension of Hoya's floral structure and adds to the taxonomic and ecological insights regarding the genus in Peninsular Malaysia.
Lake Chini's fish are important as a pool of ichthyodiversity of the Pahang River drainage and a natural food resource for the native community nearby. However, the fish population was shrinking after a disastrous flood in 1995, and it is still a puzzle as to the cause of the decline of the fish population. This study aims to estimate the fish growth pattern using the length-weight relationship (LWR) analysis and condition factor. The study evaluated b and K values from the LWR analysis equation. B value is the slope of the LWR, which indicates the growth pattern (e.g., isometric or allometric), while K value refers to the condition factor, which shows the overall health of fish. Fishes were sampled using a gill net and identified to species level, and water quality was analysed using standard methods. Lake Chini was found to have a good to moderate water quality status, ranging from category Ato category C according to the Malaysian Lake Water Quality Standard (NLWQS). The Carlson trophic index has classified the lake water bodies into an early eutrophic status. The LWR analysis on four selected fish species showed the mean b value ranged between 2.07 and 4.02, implying negative isometric growth for most of the species and isometric and positive growth for one species, which is K. cryptopterus and C.rapasson respectively. Fish length significantly influenced weight, with coefficients of determination ranging from 69% to 98%. The K value ranged from 1.00 to 1.08, indicating that the fish is in a moderate condition of well-being. This study indicates that water quality, food sources, and habitat are not the significant factors that cause fish depletion in Lake Chini.
Keropok lekor is a popular Malaysian snack made primarily from fish meat and contains very low fibre. Okara is a soy by-product from the production of soy milk and tofu, and is often discarded as food waste. However, okara is rich in nutrients, containing 55% fibre, 30% protein, 10% fat, and other bioactive components. The incorporation of okara into the production of keropok lekor can help to increase the nutritional content and reduce the environmental impact. This study aimed to evaluate the sensory and nutritional properties of okara-containing keropok lekor. Out of 16 formulations, 5 formulations, including the control, were selected for sensory evaluation: control, formulation 1 (2.7% okara), formulation 2 (5.4% okara), formulation 3 (6.7% okara), and formulation 4 (8.1% okara). The sensory evaluation revealed that formulation 1 received the highest scores for aroma, colour, and appearance and was within the acceptable range for all of these attributes, including overall acceptance. Formulation 1 showed no significant differences in colour, appearance, taste, texture, and overall acceptability compared to the control. Nutrient content of control and formulation 1 showed that fibre (3.25 +/- 0.07 g/100 g vs 5.10 +/- 0.14 g/100 g), fat (11.05 +/- 0.21 g/100 g vs 14.10 +/- 0.57 g/100 g) and energy content (289.00 +/- 0.01 g/100 g vs 325.00 +/- 2.83 g/100 g) increased after incorporation of okara, while moisture (37.20 +/- 0.14 g/100 g vs 33.55 +/- 0.07 g/100 g) and ash (4.30 +/- 0.14 g/100 g vs 2.85 +/- 0.21 g/100 g) decreased. In conclusion, keropok lekor, which contains 2.7% okara, exhibits acceptable sensory properties, suggesting that okara can be used in food development to increase dietary fibre content.
Long-tailed macaques (Macaca fascicularis) are an endangered species that are facing increasing threats due to habitat fragmentation and limited foraging grounds. Thus, their extensive range of behaviours requires attention from research and management perspectives for their conservation. Studying the behaviour of the M. fascicularis population in Tanjung Datu National Park (TDNP), Sarawak, one of the smallest protected areas that is rich in biodiversity, is crucial for ensuring their conservation success. The purpose of this study was to examine the demographic and behavioural ecology of the long-tailed macaques in TDNP, as there is currently insufficient information available on the M. fascicularis population in the protected area. A population estimation study was conducted using a census walk from February 23rd to March 1st, 2024. The behaviour of M. fascicularis was observed using the scan sampling method during this period. The study was conducted within an area of around 11,222 m2, with a total of 181 individuals recorded from two groups. Group A (175 individuals) was recorded near the HQ area, while Group B (6 individuals) was recorded in Telok Upas. No solitary individuals were observed during the study, and due to the direct count method, no variance estimate was calculated. Present results indicated that long-tailed macaques spent most oftheirtime moving (53.79%), followed by resting (14.48%), feeding (13.44%), other activity category (7.93%), vocalisation (5.52%) and grooming (4.83%). Data from the population census and behaviour of the long-tailed macaques can help the park's management in prioritising habitat protection areas and monitoring the population trends for effective long-term conservation of the species within TDNP.
Fresh pineapple juice is widely appreciated for its numerous health benefits derived from the nutrients in the fruit. Current busy lifestyles have caused many people to prefer ready-to-drink commercial functional juices, such as probiotic-added juice. The probiotics are encapsulated to preserve their bioavailability and survivability in the acidic juice. However, consumer acceptance of this enhanced juice remains uncertain, as the heating process and the addition of encapsulated probiotics may change the juice's sensory parameters (taste, texture, and colour). Therefore, focus group discussion (FGD) sessions (n=18) were conducted to assess consumer perception and acceptance of pasteurised pineapple juice with encapsulated probiotics. The FGD sessions assessed general fruit consumption, safety concerns, and health benefits of taking probiotics. The findings revealed that 100% consumers preferred the safety provided by pasteurisation and the additional health benefits associated with the addition of encapsulated probiotics to the juice. The acceptance of this experimental juice formulation indicates its potential for commercial applications.
Aromatic rice is a premium rice variety due to multiple distinctive essential qualities viz., strong aroma, long grain, intermediate amylose content, and intermediate gelatinization temperature. The strong aroma is majorly contributed by 2-acetyl-1-pyrroline (2AP), a volatile organic compound that is 10-100% higher in aromatic rice. The long grain represents a large endosperm that contains more starch and nutrients. Intermediate amylose content in aromatic rice is preferred as the rice becomes moist and tender upon cooking. As for the gelatinization temperature (GT), aromatic rice has an intermediate GT which defines a shorter time in cooking. In the world, major aromatic rice cultivars include Basmati (India & Pakistan) and Khao Dawk Mali 105 (Thailand). The production of the aforementioned varieties as well as other minorities is declining due to multiple constraints. The demand-export clash, climate change, disease outbreaks, and other abiotic factors have proven challenging for all rice industrial players. Several breeding techniques are practiced in combatting those problems. In the past, conventional pure line breeding had proven to initiate the cultivation of Basmati-370 (India) and Khao Dawk Mali 105 (Thailand). However, hybrid breeding (combination of desirable traits) is a much more effective breeding method through the release of Pusa Basmati-1 (India), Lateefy (Pakistan), and MRQ104 (Malaysia). From the genetic perspective, quantitative traits locus (QTL) mapping is used to assist in the breeding of aromatic rice cultivars. QTL mapping successfully improved yield, agronomic traits, and stress resistance by providing a strong foundation for advanced in-depth breeding technologies including marker-assisted selection (MAS), mutagenesis, and CRISPR/Cas9 mediated genome editing.
Acute myeloid leukaemia (AML) is a severe bone marrow malignancy with a high mortality rate. Recent advancements in highthroughput technology and bioinformatics have facilitated the identification of microRNAs (miRNAs) involved in the pathogenesis of AML. This study aimed to investigate the potential role of MicroRNA-let-7i-5p (miR-let-7i-5p) in the progression of AML. MiRlet-7 family has been widely utilised as a tumour inhibitor, suppressing its expression in several types of human cancer. Hence, dysregulation of miR-let-7i-5p is critical to cancer progression. However, the role of miR-let-7i-5p in AML remains unclear. Using the Gene Expression Omnibus (GEO) database, the expression of miR-let-7i-5p in AML patients was investigated. The role of miR-let-7i-5p and its potential target genes in AML was examined using bioinformatic tools. Subsequently, the effect of miR-let-7i5p was assessed through transfection, followed by the analysis of target gene expression using RT-qPCR. Further investigations employed the MTT and cell cycle assays on the effects of miR-let-7i-5p on AML cell proliferation and cell cycle. According to the GEO database, miR-let-7i-5p was significantly downregulated in AML patients compared to controls. AML development has been associated with miR-let-7i-5p by KEGG pathway analysis. Bioinformatics suggested that miR-let-7i-5p targets MYC in AML cells. Transfection of miR-let-7i-5p into AML cell lines decreased MYC expression, suggesting MYC regulates AML progression. However, MTT and cell cycle assays showed no significant effect of miR-let-7i-5p on AML cell proliferation and cell cycle, indicating the potential involvement of miR-let-7i-5p in alternative pathways in AML pathogenesis. The study highlights the potential significance of miR-let-7i-5p in regulating AML progression.
Oxidative stress occurs when there is an imbalance between the production of reactive oxygen species (ROS) and an organism's ability to detoxify these harmful intermediates, posing a significant challenge for aquatic organisms, including microalgae. Tetraselmis chui has developed defence mechanisms to manage oxidative stress, ensuring its survival and functionality in various ecosystems. This study explores the interactions between the growth, oxidative stress markers and the activation of enzymatic antioxidative defence systems in T. chui after 8 days of incubation. T. chui was cultured in F2 media and subjected to variations in media strength (F/2, F, F/4 and F/8), sucrose concentrations (0 to 50 g/L), pH (5 to 9), and light intensities (0 to 600 lux). Growth was highest in full-strength media and improved with 40 g/L sucrose addition. Growth was largely unaffected by pH, except at pH 5, which reduced it, while higher light intensities boosted fresh and dry weights. Full-strength media led to significant MDA and H2O2 accumulation, while F/2 media exhibited higher ion leakage. The absence of sucrose increased H2O2 and ion leakage but lowered MDA, whereas higher sucrose concentrations raised MDA while reducing ion leakage. pH 7 stimulated MDA production, with H2O2 concentrations largely unchanged except in controls. Cells grown in darkness showed elevated MDA and ion leakage but reduced H2O2. CAT and APX activities were significantly enhanced in F/8 media, while gPOD was highest in full-strength media. Sucrose-free media induced CAT and APX but reduced gPOD activity. All enzyme activities were most strongly induced at pH 9. Light intensity at 150 lux favoured CAT and gPOD, whereas APX was more active in dark conditions. These findings unveil the strategies employed by T. chui in response to oxidative stress, highlighting its ecological adaptability and contributing to its potential to produce antioxidants for various industrial applications.
Food adulteration remains a critical issue, particularly in highly processed foods where DNA degradation reduces the effectiveness of species identification. This study evaluated the performance of three different lengths of species-specific primers (398, 288 & 149 bp) targeting the mitochondrial cytochrome B gene in pork and aimed to determine the most effective primer for detecting pork DNA in food seasoning products using species-specific Polymerase Chain Reaction (PCR). Furthermore, species-specific primers for bovine and chicken were applied as controls to confirm species identification. The DNA was extracted from raw meat, binary mixtures, and seasonings. Sensitivity was tested with low pork concentrations, and the applicability of pork-specific primers was further evaluated in seasoning products without halal certification to assess potential pork adulteration. The results show that the DNA extracted from raw meat samples exhibited high purity (OD260/280:1.82 -2.00) and concentration (114.83 to 257.16 ng/mu L), whereas food seasoning products yielded extremely lower values of purity (OD260/280:0.97 - 1.81) and concentration (2.75 to 66.18 ng/mu L). Despite DNA degradation in processed foods, PCR products remain detectable. The shortest 149 bp primer demonstrated the highest sensitivity among the primers, successfully detecting pork DNA at a minimum concentration of 1% (w/w) in a binary meat mixture. The results demonstrated the absence of pork DNA in all non-pork labelled samples. However, the detection of chicken DNA in fermented pork cube samples indicated potential mislabelling or crosscontamination during processing. These findings emphasise the importance of primer selection for detecting highly degraded DNA. This also underscores the applicability of species-specific PCR as a practical and robust approach for routine food authentication, providing a valuable tool to ensure halal compliance within the food industry.
The fall armyworm (Spodoptera frugiperda) is a notorious invasive pest of global economic concern, impacting a diverse array of crops, with a significant preference for maize. Its swift dispersal and capacity to adapt to various environments present substantial obstacles to consistent crop yields and global food supplies. Rearing S. frugiperda on natural maize-based diets in laboratory studies is costly, space-dependent, labour-intensive, and prone to microbial contamination, underscoring the need for efficient, consistent, and less resource-demanding alternatives such as artificial diets. This study aimed to compare the developmental performance of S. frugiperda larvae reared on a natural maize stem diet versus a formulated artificial diet under controlled laboratory conditions (25 +/- 1 degrees C; 65 +/- 5% relative humidity). Larvae were subjected to two dietary treatments with three replicates each. Results showed that larvae fed the artificial diet consumed significantly more food (Mean +/- SD: 587.6 +/- 140.2 mg per larva) than those on the natural diet (432.7 +/- 179.3 mg; p<0.05). This enhanced nutritional intake reduced the life cycle to 32 days, compared to 41 days on the natural diet. Although both groups exhibited 100% larval survival, pupal weights were significantly higher in the artificial diet group (190.7 +/- 29.8 mg) than in the natural diet group (138.3 +/- 31.9 mg). The artificial diet also led to markedly lower microbial contamination (5-10%) relative to the natural diet (20-30%). Furthermore, the artificial diet group achieved 100% adult emergence. The resulting adults successfully mated and produced viable eggs, leading to successful larval development. These findings highlight the advantages of artificial diets in ensuring consistent, efficient, and contamination-reduced rearing of S. frugiperda, supporting their application in laboratory-based insect toxicology, biological control studies, and mass-rearing initiatives.
Bird's nest drinks are widely recognised as a healthy beverage option. Introducing a bird's nest orange drink represents a promising advancement in food technology, anticipated to offer enhanced health benefits. This study focuses on retort sterilisation, a crucial method for ensuring food safety and quality. However, prolonged retort processing can negatively impact beverage characteristics. This study investigated the effects of varying retort times (3, 5, and 7 minutes at 121 degrees C) and cooling methods (room temperature and ice bath) on bird's nest orange drink properties. The main goal was to find the best processing conditions to maintain product quality. Results showed that retort time significantly (p < 0.05) impacted most physical and chemical traits, including moisture, protein, fat, pH, total soluble solids, viscosity, and colour (L* values). Longer retort times decreased (p < 0.05) these properties, while ash and carbohydrate content, and redness/yellowness (a* and b* values), increased (p < 0.05). Cooling methods had less impact, mainly affecting (p < 0.05) viscosity and lightness, with ice water cooling performing slightly better. Crucially, the drink's ability to scavenge free radicals notably (p < 0.05) declined with longer retort times, regardless of the cooling method. This shows that heat treatment degrades the antioxidants. The findings highlight a trade-off between ensuring microbial safety and preserving the drink's nutritional and sensory quality, emphasising the importance of optimising thermal processing parameters for bird's nest orange drinks.
Proprotein convertase subtilisin/kexin type 9 (PCSK9) modulates cholesterol homeostasis by targeting low-density lipoprotein receptor (LDLR) for lysosomal degradation. Genetic polymorphisms in PCSK9 can alter its autocatalytic processing, secretion, or binding affinity to LDLR. Reduce binding efficiency between PCSK9 and LDLR leads to elevated low-density lipoprotein cholesterol (LDL-C) level, thereby promoting atherosclerotic plaque formation and increasing the risk of ischemic stroke. The objective of this study was to identify the most functionally significant non-synonymous single-nucleotide polymorphisms (nsSNPs) in PCSK9 via an integrated in silico analysis combining functional prediction tools (PROVEAN, SIFT, PolyPhen-2, SNAP2), protein stability and disease-association predictors, ligand-binding assessment, and post-translational modification analysis. A total of 4,979 PCSK9 variants were retrieved from Ensembl GRCh37/hg19, and HGMD. Functional annotation using PROVEAN, SIFT, PolyPhen-2, and SNAP2 identified 253 nsSNPs, with PolyPhen-2 predicting the largest subset. Upon filtering through the protein stability, disease association, ligand binding, and post-translational modification, five nsSNPs (W156R, H226L, H229R, G337R, and G394V) emerged as the most deleterious, with potential to disrupt secondary autocatalytic processing and significantly impair LDLR-PCSK9 interactions. These findings highlight novel candidate variants that may serve as diagnostic biomarkers and therapeutic targets in dyslipidemia and cardiovascular disease.
Advanced glycation end-products (AGEs) are formed through non-enzymatic reactions between reducing sugars and proteins, accelerated by high-temperature food processing. Excessive dietary intake of AGEs has been linked to chronic diseases such as diabetes, cardiovascular disorders, and neurodegeneration. An AGE content database is lacking for Malaysian foods, unlike database developments in the United States and China. This pilot study aimed to establish a protocol for AGE quantification using liquid chromatography-mass spectrometry (LC-MS) in selected Malaysian foods. Eleven food samples, categorised by processing level according to the Nova classification, were analysed for two major AGE markers - N epsilon-carboxymethyllysine (CML) and N epsilon-carboxyethyllysine (CEL). Results showed significant variation in AGE content between samples, with the highest CEL levels found in deep-fried fish balls and steamed grouper fish, while pan-fried chicken sausages had the highest CML concentration; whereas levels in fresh fruits and minimally processed items like rolled oats were below detection. These findings suggest that food processing methods, particularly dry-heat techniques, significantly influence AGE formation. This developed protocol will be applied to a larger number of Malaysian foods to facilitate the development of an AGE food composition reference database for supporting dietary guidance in chronic disease prevention strategies.
