Human Leukocyte Antigen-G Gene Polymorphism in Peninsular Malaysia: A Preliminary Report.

Introduction: Expression of the nonclassical human leukocyte antigen (HLA)-G gene is upregulated in placenta during pregnancy. In other cells, HLA-G is upregulated during parasitic infections and allergic reactions. Polymorphism at the HLA-G gene locus has been reported for many populations, but so far not for any ethnic groups in Malaysia. In this survey, we screened for genetic variation in HLA-G genes from representative Malay, Chinese, and Indian individuals living in Peninsular Malaysia. Materials and Methods: Blood samples were obtained with informed consent, and ethnicity classes were assigned based on self-declared pedigree information. Exons 2, 3, and 4 of the HLA-G gene were amplified by polymerase chain reaction and subjected to Sanger sequencing. Results: The most common genotype in Malays and Indians was found to be HLA-G*01:01:01:01/01:01:01:01 with frequencies of 0.206 and 0.167, respectively, whereas the HLA-G*01:01:03:01/01:01:01:01 genotype was the one most frequently observed in Chinese (0.221). Based on this study, HLA-G*01:01:01:01 (0.427-0.448) is the most frequent HLA-G allele in the all three ethnic groups. In contrast, HLA-G*01:01:02:01 (0.186) was observed as the second most frequent HLA-G allele in Malays and HLA-G*01:04:01 in Chinese and Indians, (0.188-0.198, respectively). Several minor HLA-G alleles were detected at low frequency in Malays, Chinese, or Indians (HLA-G*01:01:05, 01:01:09, 01:04:02, and 01:04:03). These have only rarely, if ever, been reported in other population groups. Subsequent statistical analysis including using principal coordinate data mapping showed the Malays, Chinese, and Indians are distinct but quite closely related to one another as compared with other population groups from across Europe and Africa. Conclusion: The HLA-G population data collected in this study showed that the ancestrally unrelated Malays, Chinese, and Indians are genetically distinct. This new database provides a foundation for further studies to capture HLA-G allelic diversity in uncharacterized populations of Malaysia and for future attempts to identify their roles in disease resistance and susceptibility.

Comparative analysis of water quality index and river classification in Kereh River, Penang, Malaysia: Impact of untreated swine wastewater from Kampung Selamat pig farms.

The Kereh River in Penang, Malaysia, has faced severe pollution for over 40 years due to untreated wastewater from swine farms in Kampung Selamat, discharged via stormwater drains. Despite official claims that all 77 swine farms treat their wastewater to meet regulatory standards, local non-governmental organizations and villagers have challenged this, though their concerns lack scientific backing. This study evaluates the river's water quality by analyzing samples from upstream (US), midstream (MS), and downstream (DS), and from Parit Cina-Parit Besar, a conduit for untreated swine wastewater. Fourteen parameters were measured against Malaysia's National Water Quality Standards (NWQS). Significant differences were found in six parameters: ammonium nitrogen (AN), biochemical oxygen demand (BOD), chemical oxygen demand (COD), dissolved oxygen (DO), total suspended solids (TSS), and oil and grease (OG). While Dunn's post hoc pairwise comparison showed no significant differences among river segments, mean values indicated increased pollution downstream, particularly after the convergence with untreated swine wastewater. River classification worsened, with water quality index dropping from 69.88 ± 11.37 score (Class III) US to 38.49 ± 12.74 and 50.44 ± 3.14 scores (Class IV) MS and downstream, respectively. A significant positive correlation between E. coli and AN (r = 0.71, p < 0.01) suggests a common point source pollutant, particularly the untreated swine wastewater. The river exhibits low oxygen levels and high organic matter and nutrient concentrations, especially MS and downstream, highlighting substantial ecological and public health risks. Effective enforcement of waste treatment regulations and enhanced monitoring are crucial for mitigating pollution and restoring the river's ecosystem. Collaboration between authorities and pig farmers is essential to improve water quality and maintain the river's ecological balance. PRACTITIONER POINTS: Severe Kereh River pollution: Untreated swine wastewater from Kampung Selamat pig farms, primarily via Parit Cina-Parit Besar, has degraded the river for over 40 years. Regulatory non-compliance: Despite official claims, untreated swine wastewater continues to pollute the river, challenging regulatory standards. Significant pollution indicators: Elevated levels of AN, BOD, COD, DO, TSS, OG, and E. coli signal severe pollution midstream and downstream. Water quality index drop: WQI scores classify midstream and downstream sections as polluted, indicating worsening conditions downstream. Urgent need for action: Enforcing regulations, improving wastewater treatment, and relocating pig farms are crucial for restoring the Kereh River.

Heavy metal contamination in Sungai Petani, Malaysia: a wastewater-based epidemiology study.

The aim of this study was to investigate the use of wastewater-based epidemiology (WBE) to estimate heavy metal exposure in Sungai Petani, Malaysia. Atomic absorption spectroscopy was used to detect copper (Cu), nickel (Ni), zinc (Zn), iron (Fe), and cadmium (Cd) in wastewater from eight sewage treatment facilities in Sungai Petani in January 2022. The heavy metal concentrations were measured in both influent and effluent, and the mean concentrations in the wastewater were found to be in the following order: Fe > Ni > Zn > Cd > Cu, with a 100% detection frequency. The results of WBE estimation showed that Fe, Ni, and Zn had the highest estimated per population exposure levels, while Cd had the lowest. Compared to a similar study conducted in Penang, Malaysia, all metals except Cu were found to have higher concentrations in Sungai Petani, even though it is a non-industrial district. These findings highlight the importance of addressing heavy metal contamination in Sungai Petani and implementing effective risk management and prevention strategies.

Assessment of 18 endocrine disrupting chemicals in tap water samples from Klang Valley, Malaysia.

Multiclass of endocrine disrupting chemicals (EDCs) such as nine perfluoroalkyl and polyfluoroalkyl substances (PFAS), five bisphenols, and four parabens were analysed in tap water samples from Malaysia's Klang Valley region. All samples were analysed using liquid chromatography mass tandem spectrometry (LC-MS/MS) with limit of quantitation (LOQ) ranged between 0.015 and 5 ng/mL. Fifteen of the 18 EDCs were tested positive in tap water samples, with total EDC concentrations ranging from 0.28 to 5516 ng/L for all 61 sampling point locations. In a specific area of the Klang Valley, the total concentration of EDCs was found to be highest in Hulu Langat, followed by Sepang, Putrajaya, Petaling, Kuala Lumpur, Seremban, and Gombak/Klang. PFAS and paraben were the most found EDCs in all tap water samples. Meanwhile, ethyl paraben (EtP) exhibited the highest detection rate, with 90.2% of all locations showing its presence. Over 60% of the regions showed the presence of perfluoro-n-butanoic acid (PFBA), perfluoro-n-hexanoic acid (PFHXA), perfluoro-n-octanoic acid (PFOA), perfluoro-n-nonanoic acid (PFNA), and perfluoro-1-octanesulfonate (PFOS), whereas the frequency of detection for other compounds was less than 40%. The spatial distribution and mean concentrations of EDCs in the Klang Valley regions revealed that Hulu Langat, Petaling Jaya, and Putrajaya exhibited higher levels of bisphenol A (BPA). On the other hand, Kuala Lumpur and Sepang displayed the highest mean concentrations of PFBA. In the worst scenario, the estimated daily intake (EDI) and risk quotient of some EDCs in this study exceeded the acceptable daily limits recommended by international standards, particularly for BPA, PFOA, PFOS, and PFNA, where the risk quotient (RQ) was found to be greater than 1, indicating a high risk to human health. The increasing presence of EDCs in tap water is undoubtedly a cause for concern as these substances can have adverse health consequences. This highlights the necessity for a standardised approach to evaluating EDC exposure and its direct impact on human populations' health.

A genetic Study of the Ghanaian Population Using 15 Autosomal STR Loci.

Autosomal short tandem repeat (STR) population data collected from a well characterized population are needed to correctly assigning the weight of DNA profiles in the courtroom and widely used for ancestral analyses. In this study, allele frequencies for the 15 autosomal short tandem repeat (STR) loci included in the AmpFlSTR® Identifiler® plus kit (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, FGA) were obtained by genotyping 332 unrelated individuals of Ghanaian origin. Statistical tests on STR genotype data showed no significant departure from Hardy-Weinberg equilibrium (HWE). The overall match probability, combined power of exclusion and combined power of discrimination for these loci were 1 in 3.85 × 1017, 0.99999893 and 0.99999998, respectively. Polymorphic information content (PIC) greater than 0.70 was observed for all loci except TH01 and D13S317. These statistical parameters confirm that this combination of loci is valuable for forensic identification and parentage analysis. Our results were also compared with those for 20 other human populations analyzed for the same set of markers. We observed that the Ghanaian population grouped with other African populations in two-dimensional principal coordinate (PCO) and a neighbor-joining (N-J) data mapping and placed closest to Nigerians. This observation reflects cultural similarities and geographical factors, coupled with the long history of migration and trading activities between Ghana and Nigeria. Our report provides what we believe to be the first published autosomal STR data for the general Ghanaian population using 15 loci genotyped using the AmpFlSTR® Identifiler® plus kit methodology. Our data show that the loci tested have sufficient power to be used reliably for DNA profiling in forensic casework and help to elucidate the genetic history of people living in the country.

PFAS, bisphenol, and paraben in Malaysian food and estimated dietary intake.

Endocrine disrupting chemicals (EDCs) such as per- and polyfluoroalkyl substances (PFAS), bisphenols, and parabens are used in food packaging or as preservatives and their unintended consumption has been associated with cancer and other diseases. Food EDCs data are scarce in Malaysia. Thus, liquid chromatography mass tandem spectrometry (LC-MS/MS) was utilised to analyse 18 EDCs from different food categories. Bisphenol was the most abundant EDC found, followed by PFAS and paraben. Bisphenol levels in canned foods, dairy products, canned drinks, fruits, and vegetables ranged from 1.16 to 183 ng/g. PFAS was found in almost every food category, with canned foods having the highest concentrations (0.18-34.5 ng/g). Only canned foods, fruits, and vegetables contained parabens, with mean concentrations ranging from 0.27 to 26.7 ng/g. PFOS, PFBA, PFHQA and bisphenol A all had hazard quotients (HQ) above 1, indicating that they can pose a risk to human health.

Distribution of 22 Single Nucleotide Polymorphisms in 13 Cytokine Genes in Malays, Chinese, and Indians in Peninsular Malaysia.

Introduction: Cytokines are cell signaling glycoproteins that are particularly important in immunity and inflammatory responses. Therefore, variations, such as single nucleotide polymorphisms (SNPs), in genes encoding for cytokines may have important consequences for their roles in health. Materials and Methods: A total of 222 unrelated, healthy, and un-admixed Malays (n = 97), Chinese (n = 77), and Indians (n = 48) with a median age of 30 years old (range 21-50) were typed for 22 cytokine gene SNPs: IL-1α -889 T/C, IL-1ß (-511 T/C, +3962 T/C), IL-1R pst1 1970 T/C, IL-1RA mspa1 11100 T/C, IL-4Rα +1902 G/A, IL-12 - 1188 C/A, IFN-γ +874 A/T, TGF-ß (cdn 10 C/T, cdn 25 G/C), TNF-α (-308 A/G, -238 A/G) IL-2 (+166 G/T, -330 T/G), IL-4 (-1098 T/G, -590 T/C, -33 T/C), IL-6 (-174 C/G, nt565 G/A), and IL-10 (-1082 G/A, -819 C/T, -592 A/C). This involved using well-established polymerase chain reaction procedures with sequence-specific primers and restriction fragment length polymorphism methods. Results: The majority of the screened cytokine gene SNPs are polymorphic in all three ethnicities. Exceptions include TGF-ß cdn 25 (G/C), IL-1ß +3962 (T/C), and TNF-α -238 (A/G), which were all observed to be monomorphic in Malays, Chinese and Indians. Many of the analyzed cytokine gene SNP genotypes deviated from Hardy-Weinberg equilibrium and the three ethnic study groups were all well-separated from reference Asian, African and European populations in a principal component analysis plot. Conclusion: We successfully typed 22 SNPs in 13 cytokine genes from genetic material collected from unrelated and un-admixed Malay, Chinese and Indian individuals in Peninsular Malaysia. These new cytokine gene population datasets reveal interesting contrasts with other populations.

Heavy Metal Concentrations in Malaysian Adults' Hair and Associated Variables in Bukit Mertajam, Penang, Malaysia.

The presence of heavy metals in human hair is being tracked to predict health risk, forensics, and environmental monitoring. Heavy metals are typically non-biodegradable and have a lengthy half-life, allowing them to linger in humans and the environment for many years. Heavy metal exposure in hair has been attributed to multiple sources from the environment and food intake. In this study, copper (Cu), nickel (Ni), zinc (Zn), chromium (Cr), manganese (Mn), lead (Pb), and cadmium (Cd) levels were measured in the scalp hair of 50 individuals in Bukit Mertajam, Penang, Malaysia. In conjunction with sampling, subjects' age, gender, lifestyle, diet, and working environment were also obtained through the questionnaire. The Atomic Absorption Spectrometry (AAS) method was used to extract all the metals in the hair samples. The mean concentrations of heavy metals were found to be in the following order (unit of mg/kg): Cr > Zn > Pb > Ni > Cd > Cu. Manganese was detected below the limit of quantitation among the elements (< LOQ). All elements except Mn were higher and comparable to the previous studies' international limit values. Cadmium prevalence was substantially associated with age, smoking habit, dyed hair, and working environment in Pearson's correlation analysis (p ≤ 0.05). Zinc was also found to be related to the working environment. Some elements were observed to be statistically related between heavy metals, Cd/Zn, Cd/Ni, Cr/Ni, and Pb/Ni, whereas smoking habit/dyed hair and dyed hair/working environment were the associated factors for metal distribution that were statistically correlated (p ≤ 0.05). To recapitulate, this study found that the distribution of heavy metals in hair was influenced by associated factors and between heavy metals. It has been indicated that heavy metal exposure to humans is influenced by factors such as geographical location, lifestyle, and working environment.

In vitro Anticancer Effects of Vernonia amygdalina Leaf Extract and Green-Synthesised Silver Nanoparticles.

PURPOSE: Vernonia amygdalina (VA) is a traditional African herbal medicine that has been reported to possess anticancer properties. However, the anticancer properties of VA silver nanoparticles have not been studied. The aim of the study was to examine and evaluate the anticancer activities of VA leaf extracts and VA silver nanoparticles on the human breast cancer cell line, MCF-7. METHODS: VA leaves were extracted using sequential extraction assisted with ultrasound using three different solvents: ethanol, 50% ethanol, and deionized water. The silver nanoparticles were synthesised with VA aqueous extract. RESULTS: The ethanol extract and VA silver nanoparticles inhibit MCF-7 cell proliferation with an average half-maximal inhibitory concentration (IC50) value of 67µg/mL and 6.11µg/mL, respectively, after 72 hours of treatment. The ethanol extract and VA silver nanoparticles also caused G1 phase cell cycle arrest, induced apoptosis and nuclear fragmentation in MCF-7 cells. CONCLUSION: VA ethanol extracts and VA silver nanoparticles decreased the cell viability in MCF-7 cells in a time and dose-dependent manner by inducing apoptosis and causing DNA damage. Further research is needed to elucidate the mechanism of action of VA leaf extracts and VA silver nanoparticles. This study is the first to report on the anticancer activity of VA silver nanoparticles in MCF-7 cells.

Population data and genetic characteristics of 12 X-STR loci using the Investigator® Argus X-12 Quality Sensor kit for the Kedayan population of Borneo in Malaysia.

DNA profiling of X-chromosomal short tandem repeats (X-STR) has exceptional value in criminal investigations, especially for complex kinship and incest cases. In this study, Investigator® Argus X-12 Quality Sensor (QS) kits were successfully used to characterize 12 X-STR loci in 199 unrelated healthy Kedayan individuals living in Sabah and Sarawak, Malaysia. The LG1 haplogroup (DXS8378 - DXS10135 - DXS10148) has the largest HD (0.9799) as compared with all other closely linked haplotype groups examined (LG2; DXS7132-DXS10074-DXS10079, LG3; DXS10103-DXS10101-HPRTB and LG4; DXS10134-DXS7423-DXS10146). Data from statistical analysis showed that high combined of PDM, PDF, MEC_Krüger, MEC_Kishida, MEC_Desmarais, and MEC_Desmarais_duo values (0.999999994405922, 0.99999999999999, 0.999990463834938, 0.999999975914808, 0.999999975985006, and 0.999996491927194, respectively) in the Kedayan. In a two-dimensional scaling (MDS) plot and dendrogram constructed using allele frequencies at the 12 X-STR loci, Kedayan appear to be most closely related to their other Austronesian populations including the Malays and Filipinos as compared with other reference population groups. Findings from the present study thus demonstrate high genetic variability across the 12 tested X-STR loci and can be used for population studies and forensic applications.

A simple and efficient sequential electrokinetic and hydrodynamic injections in micellar electrokinetic chromatography method for quantification of anticancer drug 5-fluorouracil and its metabolite in human plasma.

A simple and sensitive preconcentration strategy using sequential electrokinetic and hydrodynamic injection modes in micellar electrokinetic chromatography with diode array detection was developed and applied for the separation and determination of anticancer agent, 5-fluorouracil and its metabolite, 5-fluoro-2'-deoxyuridine, in human plasma. Sequential injection modes with increased analyte loading capacity using the anionic pseudo-stationary phase facilitated collection of the dispersed neutral and charged analytes into narrow zones and improved sensitivity. Several important parameters affecting sample enrichment performance were evaluated and optimized in this study. Under the optimized experimental conditions, 614- and 643-fold and 782- and 803-fold sensitivity improvement were obtained for 5-fluorouracil and its metabolite when compared with normal hydrodynamic and electrokinetic injection, respectively. The method has good linearity (1-1,000 ng/ml) with acceptable coefficient of determination (r2 > 0.993), low limits of detection (0.11-0.14 ng/ml) and satisfactory analyte relative recovery (97.4-99.7%) with relative standard deviations of 4.6-9.3% (n = 6). Validation results as well as the application to analysis of human plasma samples from cancer patients demonstrate the applicability of the proposed method to clinical studies.

Pharmacokinetics and Biodistribution of Thymoquinone-loaded Nanostructured Lipid Carrier After Oral and Intravenous Administration into Rats.

BACKGROUND: Thymoquinone (TQ), an active compound isolated from Nigella sativa, has been proven to exhibit various biological properties such as antioxidant. Although oral delivery of TQ is valuable, it is limited by poor oral bioavailability and low solubility. Recently, TQ-loaded nanostructured lipid carrier (TQ-NLC) was formulated with the aim of overcoming the limitations. TQ-NLC was successfully synthesized by the high-pressure homogenization method with remarkable physiochemical properties whereby the particle size is less than 100 nm, improved encapsulation efficiency and is stable up to 24 months of storage. Nevertheless, the pharmacokinetics and biodistribution of TQ-NLC have not been studied. This study determined the bioavailability of oral and intravenous administration of thymoquinone-loaded nanostructured lipid carrier (TQ-NLC) in rats and its distribution to organs. MATERIALS AND METHODS: TQ-NLC was radiolabeled with technetium-99m before the administration to the rats. The biodistribution and pharmacokinetics parameters were then evaluated at various time points. The rats were imaged at time intervals and the percentage of the injected dose/gram (%ID/g) in blood and each organ was analyzed. RESULTS: Oral administration of TQ-NLC exhibited greater relative bioavailability compared to intravenous administration. It is postulated that the movement of TQ-NLC through the intestinal lymphatic system bypasses the first metabolism and therefore enhances the relative bioavailability. However, oral administration has a slower absorption rate compared to intravenous administration where the AUC0-∞ was 4.539 times lower than the latter. CONCLUSION: TQ-NLC had better absorption when administered intravenously compared to oral administration. However, oral administration showed greater bioavailability compared to the intravenous route. This study provides the pharmacokinetics and biodistribution profile of TQ-NLC in vivo which is useful to assist researchers in clinical use.

Population dataset for 21 simple tandem repeat loci in the Akan population of Ghana.

Short tandem repeat (STR) loci are widely used as genetic marker for ancestral and forensic analyses. The latter application includes for paternity testing and DNA profiling of samples collected from scenes of crime and suspects. This survey provides the first dataset for 21 STR loci across the Akan population in Ghana by genotyping of 109 unrelated healthy individuals using Investigator 24plex kit. None of the STR loci screened deviated from Hardy-Weinberg equilibrium after applying Bonferroni correction. Overall, 224 unique alleles were observed with allele frequencies ranging from 0.005 to 0.518. The combined match probability, combined power of exclusion and combined power discrimination were 1 in 4.07 × 10-25, 0.999999999 and 1, respectively. Principal coordinate analysis carried out using 21 STR allele frequency data mapped the Akans with Nigerian subpopulation groups (Hausa, Igbo and Yoruba), but separated from Thais of Thailand, Chechen of Jordan and Tijuana of Mexico.

Liquid Chromatographic Tandem Mass Spectrometric (LC-MS/MS) Determination of Perfluorooctane Sulfonate (PFOS) and Perfluorooctanoic Acid (PFOA) in the Yolk of Poultry Eggs in Malaysia.

Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) are widely used in products, and are known for their water and grease repellent properties. The persistence nature and potential toxicity of these substances have raised substantial concerns about health effects. Regarding humans, food consumption has reportedly been a significant source of exposure for both compounds. Hence, this study was performed to develop and validate an analytical method for PFOS and PFOA in egg yolks using liquid chromatographic tandem mass spectrometry (LC-MS/MS) followed by the determination of concentration of both compounds in the yolk of poultry eggs in Malaysia. A total of 47 poultry egg yolk samples were extracted by a simple protein precipitation technique using acetonitrile. The analytical method was developed using LC-MS/MS and validated based on the Food and Drug Administration (FDA)'s Bioanalytical Method Validation guidelines. The results revealed that PFOS was quantitatively detected in six samples, with the concentration range between 0.5 and 1.01 ng g-1. Among these, five samples were from home-produced chicken eggs, and one sample was from a quail egg. The levels of PFOA in all samples were below the quantifiable limit (<0.1 ng g-1). This indicated that the contamination of PFCs in poultry eggs were mostly attributed to the nature of free foraging animals, which had direct contact with the contaminants in soil and feed. In conclusion, a fast and robust analytical method for analyzing PFOS and PFOA in egg yolk samples using LC-MS/MS was successfully developed and validated. The presence of these emerging contaminants in this study signified widespread pollution in the environment.

Inhibition of CYP3A by Antimalarial Piperaquine and Its Metabolites in Human Liver Microsomes With IVIV Extrapolation.

The potential of the antimalarial piperaquine and its metabolites to inhibit CYP3A was investigated in pooled human liver microsomes. CYP3A activity was measured by liquid chromatography-tandem mass spectrometry as the rate of 1'-hydroxymidazolam formation. Piperaquine was found to be a reversible, potent inhibitor of CYP3A with the following parameter estimates (%CV): IC50 = 0.76 µM (29), Ki = 0.68 µM (29). In addition, piperaquine acted as a time-dependent inhibitor with IC50 declining to 0.32 µM (28) during 30-min pre-incubation. Time-dependent inhibitor estimates were kinact = 0.024 min-1 (30) and KI = 1.63 µM (17). Metabolite M2 was a highly potent reversible inhibitor with estimated IC50 and Ki values of 0.057 µM (17) and 0.043 µM (3), respectively. M1 and M5 metabolites did not show any inhibitory properties within the limits of assay used. Average (95th percentile) simulated in vivo areas under the curve of midazolam increased 2.2-fold (3.7-fold) on the third which is the last day of piperaquine dosing, whereas for its metabolite M2, areas under the curve of midazolam increased 7.7-fold (13-fold).

LC-MS/MS quantitation of antimalarial drug piperaquine and metabolites in human plasma.

PURPOSE: This study aimed to develop a sensitive, quantitative assay for the antimalarial piperaquine (PQ) and its metabolites M1 and M2 in human plasma. RESULTS: Analytes were gradiently separated on a C18 column and detected with a Sciex API 4000 MS/MS with an ESI source operated in the positive ion mode with deuterated PQ as internal standard. The response was linear in the range 3.9-2508nM with a runtime of 7.0min per sample. The method was applied to clinical samples from healthy volunteers. CONCLUSION: This LC-MS/MS method for the simultaneous quantitation of PQ and two of its metabolites in plasma may prove helpful for assessment of metabolite safety issues in vivo.