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.

Adaptive natural killer cell expression in response to cytomegalovirus infection in blood and solid cancer.

Natural Killer (NK) cells are conventionally thought to be an indefinite part of innate immunity. However, in a specific subset of NK cells, recent data signify an extension of their "duties" in immune surveillance and response, having characteristics of adaptive immunity, in terms of persistence and cytotoxicity. These cells are known as the adaptive or memory-like NK cells, where human cytomegalovirus (HCMV) infection has been shown to drive the expansion of adaptive NKG2C+ NK cells. HCMV is a ubiquitous pathogen whose prevalence differs worldwide with respect to the socioeconomic status of countries. The adaptive NK cell subpopulation is often characterized by the upregulated expression of NKG2C, CD16, and CD2, and restricted expression of NKG2A, FCεRγ and killer immunoglobulin-like receptors (KIR), although these phenotypes may differ in different disease groups. The reconfiguration of these receptor distributions has been linked to epigenetic factors. Hence, this review attempts to appraise literature reporting markers associated with adaptive or memory-like NK cells post-HCMV infection, in relation to solid cancers and hematological malignancies. Adaptive NK cells, isolated and subjected to ex vivo modifications, have the potential to enhance anti-tumor response which can be a promising strategy for adoptive immunotherapy.

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.

The emerging roles of interstitial macrophages in pulmonary fibrosis: A perspective from scRNA-seq analyses.

Pulmonary fibrosis is an irreversible and progressive disease affecting the lungs, and the etiology remains poorly understood. This disease can be lethal and currently has no specific clinical therapeutic regimen. Macrophages, the most common type of immune cell in the lungs, have been reported to play a key role in the pathogenesis of fibrotic disease. The lung macrophage population is mostly composed of alveolar macrophages and interstitial macrophages, both of which have not been thoroughly studied in the pathogenesis of lung fibrosis. Interstitial macrophages have recently been recognised for their participation in lung fibrosis due to new technology arising from a combination of bioinformatics and single-cell RNA sequencing analysis. This paper reviews recent developments regarding lung macrophage classification and summarizes the origin and replenishment of interstitial macrophages and their function in pulmonary fibrosis.

Vibrational Spectroscopy-Based Chemometrics Analysis of Clinacanthus nutans Extracts after Postharvest Processing and Extract Effects on Cardiac C-Kit Cells.

Chemical constituents in plants can be greatly affected by postharvest processing, and it is important to identify the factors that lead to significant changes in chemistry and bioactivity. In this study, attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy was used to analyze extracts of Clinacanthus nutan (C. nutans) leaves generated using different parameters (solvent polarities, solid-liquid ratios, ultrasonic durations, and cycles of extraction). In addition, the effects of these extracts on the viability of cardiac c-kit cells (CCs) were tested. The IR spectra were processed using SIMCA-P software. PCA results of all tested parameter sets were within acceptable values. Solvent polarity was identified as the most influential factor to observe the differences in chemical profile and activities of C. nutans extracts. Ideal extraction conditions were identified, for two sample groups (G1 and G2), as they showed optimal total phenolic content (TPC) yield of 44.66 ± 0.83 mg GAE/g dw and 45.99 ± 0.29 mg GAE/g dw and CC viability of 171.81 ± 4.06% and 147.53 ± 6.80%, respectively. Validation tools such as CV-ANOVA (p < 0.05) and permutation (R 2 and Q 2 plots were well intercepted to each other) have further affirmed the significance and reliability of the partial least square (PLS) model of solvent polarity employed in extraction. Hence, these approaches help optimize postharvest processes that encourage positive TPC and CCs results in C. nutans extracts.

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.

Stimulation of Innate and Adaptive Immune Cells with Graphene Oxide and Reduced Graphene Oxide Affect Cancer Progression.

Sole nanomaterials or nanomaterials bound to specific biomolecules have been proposed to regulate the immune system. These materials have now emerged as new tools for eliciting immune-based therapies to treat various cancers. Graphene, graphene oxide (GO) and reduced GO (rGO) are the latest nanomaterials among other carbon nanotubes that have attracted wide interest among medical industry players due to their extraordinary properties, inert-state, non-toxic and stable dispersion in a various solvent. Currently, GO and rGO are utilized in various biomedical application including cancer immunotherapy. This review will highlight studies that have been carried out in elucidating the stimulation of GO and rGO on selected innate and adaptive immune cells and their effect on cancer progression to shed some insights for researchers in the development of various GO- and rGO-based immune therapies against various cancers.

Human umbilical cord-mesenchymal stem cells: a promising strategy for corneal epithelial regeneration.

Aim: As a strategy to improve the outcome of ex vivo cultivated corneal epithelial transplantation, the role of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) is investigated in promoting corneal epithelial growth and functions. Materials & methods: Human telomerase-immortalized corneal epithelial cells were characterized and its functions evaluated by scratch migration assay, cellular senescence, HLA expression and spheres formation with hUC-MSC. Results: Expression of corneal epithelial markers was influenced by the duration and method of co-culture. Indirect co-culture improved cellular migration and delayed senescence when treated after 3 and 5 days. hUC-MSC downregulated expression of HLA Class I and II in IFN-γ-stimulated human telomerase-immortalized corneal epithelial cells. Conclusion: hUC-MSC promote corneal epithelial growth and functions after treatment with hUC-MSC.

Gynura procumbens ethanol extract and its fractions inhibit macrophage derived foam cell formation.

ETHNOPHARMACOLOGY RELEVANCE: Gynura procumbens (Lour.) Merr. displayed cardio-protective effect that may prevent atherogenesis. The primary underlying pathological process of cardiovascular disease is atherosclerosis. Atherosclerotic lesion composed of macrophages, T cells and other immune cells which incorporated with cholesterol that infiltrates from the blood. AIM OF THE STUDY: The present study was performed to determine underlying mechanism of G. procumbens ethanol extract and its fractions such as aqueous, chloroform, ethyl acetate and hexane affect macrophage derived foam cell formation. MATERIALS AND METHODS: Lipid droplets accumulation in treated macrophages were visualized by Oil Red O staining while the total cholesterol present in the treated macrophages were measured using Cholestryl Ester quantification assay kit. Enzyme-Linked Immunosorbent Assay (ELISA) were used to detect TNF-α and IL-1ß secretion in the supernatant of treated macrophages. Gene expression of Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) and ATP-binding cassette transporter A-1 (ABCA-1) in treated macrophages were analyzed using Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR). RESULTS: G. procumbens ethanol extract and its fractions reduced lipid droplet accumulation and total cholesterol in oxLDL-treated macrophages together with significantly reduction of TNF-α and IL-1ß secretions in supernatant oxLDL-treated macrophages. LOX-1 gene expression was significantly reduced when G. procumbens ethanol extract and its fractions were added in oxDL-treated macrophages. In contrast, G. procumbens ethanol extract and its fractions significantly increased the expression of ABCA-1 gene in oxLDL-treated macrophages. CONCLUSION: In conclusion, G. procumbens ethanol extract and its fractions inhibit the formation of macrophage derived foam cell by reducing TNF-α and IL-1ß expression, which usually highly expressed in atherosclerotic plaques, suppressing scavenger receptor LOX-1 gene that binds oxLDL but induced ABCA-1 gene that mediate lipid efflux from macrophages.

Stimulatory Secretions of Airway Epithelial Cells Accelerate Early Repair of Tracheal Epithelium.

Airway stem/progenitor epithelial cells (AECs) are notable for their differentiation capacities in response to lung injury. Our previous finding highlighted the regenerative capacity of AECs following transplantation in repairing tracheal injury and reducing the severity of alveolar damage associated acute lung injury in a rabbit model. The goal of this study is to further investigate the potential of AECs to re-populate the tracheal epithelium and to study their stimulatory effect on inhibiting pro-inflammatory cytokines, epithelial cell migration and proliferation, and epithelial-to-mesenchymal transition (EMT) process following tracheal injury. Two in vitro culture assays were applied in this study; the direct co-culture assay that involved a culture of decellularised tracheal epithelium explants and AECs in a rotating tube, and indirect co-culture assay that utilized microporous membrane-well chamber system to separate the partially decellularised tracheal epithelium explants and AEC culture. The co-culture assays provided evidence of the stimulatory behaviour of AECs to enhance tracheal epithelial cell proliferation and migration during early wound repair. Factors that were secreted by AECs also markedly suppressed the production of IL-1ß and IL-6 and initiated the EMT process during tracheal remodelling.

Construction and Cloning of Reporter-Tagged Replicon cDNA for an In Vitro Replication Study of Murine Norovirus-1 (MNV-1).

BACKGROUND: A norovirus maintains its viability, infectivity and virulence by its ability to replicate. However, the biological mechanisms of the process remain to be explored. In this work, the NanoLuc™ Luciferase gene was used to develop a reporter-tagged replicon system to study norovirus replication. METHODS: The NanoLuc™ Luciferase reporter protein was engineered to be expressed as a fusion protein for MNV-1 minor capsid protein, VP2. The foot-and-mouth disease virus 2A (FMDV2A) sequence was inserted between the 3'end of the reporter gene and the VP2 start sequence to allow co-translational 'cleavage' of fusion proteins during intracellular transcript expression. Amplification of the fusion gene was performed using a series of standard and overlapping polymerase chain reactions. The resulting amplicon was then cloned into three readily available backbones of MNV-1 cDNA clones. RESULTS: Restriction enzyme analysis indicated that the NanoLucTM Luciferase gene was successfully inserted into the parental MNV-1 cDNA clone. The insertion was further confirmed by using DNA sequencing. CONCLUSION: NanoLuc™ Luciferase-tagged MNV-1 cDNA clones were successfully engineered. Such clones can be exploited to develop robust experimental assays for in vitro assessments of viral RNA replication.

The Role of Isocitrate Lyase (ICL1) in the Metabolic Adaptation of Candida albicans Biofilms.

BACKGROUND: A major characteristic of Candida biofilm cells that differentiates them from free-floating cells is their high tolerance to antifungal drugs. This high resistance is attributed to particular biofilm properties, including the accumulation of extrapolymeric substances, morphogenetic switching, and metabolic flexibility. OBJECTIVES: This study evaluated the roles of metabolic processes (in particular the glyoxylate cycle) on biofilm formation, antifungal drug resistance, morphology, and cell wall components. METHODS: Growth, adhesion, biofilm formation, and cell wall carbohydrate composition were quantified for isogenic Candida albicans ICL1/ICL1, ICL1/icl1, and icl1/icl1 strains. The morphology and topography of these strains were compared by light microscopy and scanning electron microscopy. FKS1 (glucan synthase), ERG11 (14-α-demethylase), and CDR2 (efflux pump) mRNA levels were quantified using qRT-PCR. RESULTS: The ICL1/icl1 and icl1/icl1 strains formed similar biofilms and exhibited analogous drug-tolerance levels to the control ICL1/ICL1 strains. Furthermore, the drug sequestration ability of ß-1, 3-glucan, a major carbohydrate component of the extracellular matrix, was not impaired. However, the inactivation of ICL1 did impair morphogenesis. ICL1 deletion also had a considerable effect on the expression of the FKS1, ERG11, and CDR2 genes. FKS1 and ERG11 were upregulated in ICL1/icl1 and icl1/icl1 cells throughout the biofilm developmental stages, and CDR2 was upregulated at the early phase. However, their expression was downregulated compared to the control ICL1/ICL1 strain. CONCLUSIONS: We conclude that the glyoxylate cycle is not a specific determinant of biofilm drug resistance.

The modulation of PPARγ1 and PPARγ2 mRNA expression by ciglitazone in CD3/CD28-activated naïve and memory CD4+ T cells.

Given their roles in immune regulation, the expression of the nuclear receptor peroxisome proliferator-activated receptor γ (PPARγ) 1 and 2 isoforms was investigated in human naïve (CD45RA+) and memory (CD45RO+) CD4+ T cells. Stimulation of both types of cells via the CD3/CD28 pathway resulted in high expression of both PPARγ receptors as measured by real-time PCR. Treatment with the PPARγ agonist, ciglitazone, increased PPARγ1 expression but decreased PPARγ2 expression in stimulated naïve and memory cells. Furthermore, when present, the magnitude of both PPARγ receptors expression was lower in naïve cells, perhaps suggesting a lower regulatory control of these cells. Similar profiles of selected proinflammatory cytokines were expressed by the two cell types following stimulation. The induction of PPARγ1 and suppression of PPARγ2 expressions in naïve and memory CD4+ T cells in the presence of ciglitazone suggest that the PPARγ subtypes may have different roles in the regulation of T-cell function.

Usage of traditional medicines among elderly and the prevalence of prednisolone contamination.

The elderly consume many medications including traditional medicines. In 1986, it was found that 29% of elderly took traditional medicines although in 1996, the National Health Morbidity survey reported a 2.3% prevalence. However, studies from other countries showed much higher percentages. The Ministry of Health in Malaysia is concerned that some of these preparations maybe contaminated with steroids, antihistamines, hormones and other poisons. The aims of the study were to determine a). the health seeking behaviour of elderly Malays living in rural areas, b). the utilization of both modern and traditional medicines and c). the steroid content of the traditional medicines used. Methodology included interviews using structured questionnaires of elderly Malays living in rural areas of Kelantan, aged above 60 years. Samples of traditional medications collected were sent to the Pharmacology Department, School of Medical Sciences, Universiti Sains Malaysia, for steroid content analysis using Thin Layer Chromatography. A total of 599 elderly respondents were interviewed comprising 62.4% females and 37.6% males. The 60-69 years cohort group made up 48.7%, followed by 70-79 years at 36.1% and the remainder 15.2% were more than 80 years. There were 82% of elderly taking medicines. The trends of utilization of modern and traditional medicine in the last two weeks among elderly were 59.3% and 40.9% respectively. The utilization of traditional medicine by rural elderly Malays was therefore much higher than that reported in the previous study and nearly similar to that of France and Australian studies. There were 102 samples of traditional medications collected and analysed for steroid content. Results showed that 27.5% were positive for prednisolone, 34.3% positive for unknown steroids (a total of 61.8%) and 38.2% were negative for both steroids. The present study therefore once again confirmed the high usage of traditional medicines where some of which are contaminated with steroids.