Browse publications by year: 2023

  1. Ch'ng ACW, Konthur Z, Lim TS
    Methods Mol Biol, 2023;2702:291-313.
    PMID: 37679626 DOI: 10.1007/978-1-0716-3381-6_15
    Bio-panning is a common process involved in recombinant antibody selection against defined targets. The biopanning process aims to isolate specific antibodies against an antigen via affinity selection from a phage display library. In general, antigens are immobilized on solid surfaces such as polystyrene plastic, magnetic beads, and nitrocellulose. For high-throughput selection, semi-automated panning selection allows simultaneous panning against multiple target antigens adapting automated particle processing systems such as the KingFisher Flex. The system setup allows for minimal human intervention for pre- and post-panning steps such as antigen immobilization, phage rescue, and amplification. In addition, the platform is also adaptable to perform polyclonal and monoclonal ELISA for the evaluation process. This chapter will detail the protocols involved from the selection stage until the monoclonal ELISA evaluation with important notes attached at the end of this chapter for optimization and troubleshooting purposes.
    MeSH terms: Antibodies; Bacteriophages*; Humans; Magnetite Nanoparticles*; Cell Surface Display Techniques; Bioprospecting
  2. Yu H, Zahidi I, Chow MF
    iScience, 2023 Sep 15;26(9):107667.
    PMID: 37680487 DOI: 10.1016/j.isci.2023.107667
    As global demand for natural resources escalates, the environmental impact stemming from resource extraction has risen to the forefront of contemporary discussions. This paper probed the potential of using vegetation cover as an ecological barometer to gauge the level of environmental damage and restoration in mining areas: a decline in vegetation cover may signify detrimental impacts from intense mining activities, while an increase may indicate effective local environmental stewardship. Therefore, this paper undertook an assessment and discussion of mining damage and environmental management at China's Ta'ershan Mining Area since 2007, calculating and visualizing FVC (Fractional Vegetation Cover) of the Ta'ershan Mining Area to track changes in vegetation cover between 2007 and 2021. Changes in vegetation cover in the Ta'ershan Mining Area could act as a reflection of both mining-induced damage and subsequent successful environmental management by local authorities, providing a practical way to evaluate ecological effects in resource development.
  3. Attiq A, Afzal S
    Front Pharmacol, 2023;14:1255727.
    PMID: 37680708 DOI: 10.3389/fphar.2023.1255727
    Unresolved inflammation is a pathological consequence of persistent inflammatory stimulus and perturbation in regulatory mechanisms. It increases the risk of tumour development and orchestrates all stages of tumorigenesis in selected organs. In certain cancers, inflammatory processes create the appropriate conditions for neoplastic transformation. While in other types, oncogenic changes pave the way for an inflammatory microenvironment that leads to tumour development. Of interest, hallmarks of tumour-promoting and cancer-associated inflammation are striking similar, sharing a complex network of stromal (fibroblasts and vascular cells) and inflammatory immune cells that collectively form the tumour microenvironment (TME). The cross-talks of signalling pathways initially developed to support homeostasis, change their role, and promote atypical proliferation, survival, angiogenesis, and subversion of adaptive immunity in TME. These transcriptional and regulatory pathways invariably contribute to cancer-promoting inflammation in chronic inflammatory disorders and foster "smouldering" inflammation in the microenvironment of various tumour types. Besides identifying common target sites of numerous cancer types, signalling programs and their cross-talks governing immune cells' plasticity and functional diversity can be used to develop new fate-mapping and lineage-tracing mechanisms. Here, we review the vital molecular mechanisms and pathways that establish the connection between inflammation and tumour development, progression, and metastasis. We also discussed the cross-talks between signalling pathways and devised strategies focusing on these interaction mechanisms to harness synthetic lethal drug combinations for targeted cancer therapy.
  4. Patwardhan B, Chaturvedi S, Chuthaputti A, Soon GC, Abeygunawardana D, Chol KS
    PMID: 37680752 DOI: 10.1016/j.lansea.2023.100270
  5. Almayahi BA, Ali AH
    Heliyon, 2023 Sep;9(9):e19591.
    PMID: 37681124 DOI: 10.1016/j.heliyon.2023.e19591
    This study aimed to develop and test a biosensor for detecting radioactive radon gas and lead ions in blood samples collected from donors in Iraq. The biosensor was made up of aptamer, acetic acid, malachite green, and TRIS-HAC, and results were measured using a fluorescence spectrophotometer. This study found that 222Rn in the blood varied between individuals, with higher levels in males and smokers, and the highest concentration found in a male patient with cancer. The biosensor used to detect 222Rn in the blood was effective, sensitive, and low-cost, and the levels detected were within the limits set by the WHO. The study also looked at pb+2, a toxic metal, and found that levels were within permissible limits. The biosensor was also effective in detecting pb+2. The correlations between the variables are generally weak to moderate, and there are some negative relationships between humidity and other variables. There are also some strong positive relationships between temperature (Tin) and temperature (Tout). The results suggest that these variables are not strongly correlated with each other, which is an important finding for understanding their potential effects on health outcomes. However, further validation and testing may be necessary before its widespread use in clinical settings. This study highlights the importance of monitoring these substances in the blood, especially for individuals with occupational exposure to radiation. The biosensor was found to be sensitive, cost-effective, fast to manufacture, and efficient compared to other detection devices. Therefore, the study recommends the use of this biosensor for measuring radon and lead ions in blood samples. The biosensor used in this study could be a useful tool for such monitoring.
  6. Al Bargi WA, Daniel BD, Khalifa NA, Rohani MM, Hussain Q, Hamdan RB
    Heliyon, 2023 Sep;9(9):e19310.
    PMID: 37681154 DOI: 10.1016/j.heliyon.2023.e19310
    A zebra crossing is a form of pedestrian crossing provision point on roads that have significant volumes of traffic. It is one of the safety measures employed to reduce avoidable pedestrian-motorist conflicts and accidents on such roads. In the past several studies have been conducted on the behaviours of road users (pedestrians and motorists) especially on non-signal intersections. Although, various recommendations and solutions have been proffered to the many road-crossing challenges. But there has been little to no change in pedestrians' perceptions and preferences of zebra crossing. Contemporary researches have tried to rectify this by applying several models to rate the use of pedestrian zebra crossing. This study uses a Regression Model Techniques to analyse factors influencing utilization rate of pedestrian zebra crossing. In this study, 450 samples were collected from 12 locations, covering school, public building, residential and business areas to examine the utilization rate of the zebra crossing by pedestrians. To examine the significance level on the crossing utilization rates by pedestrian at 95% confidence interval, a pedestrian utilization rate (PUR) was acting as the dependent variable and the remaining variables served as the independent variables. The Multiple Linear Regression (MLR) model was also used to determine the utilization rate needed to develop the zebra crossing utilization model. From the findings, the calibrated R2 value was discovered to be at 0.937 and the descriptive statistics of MLR test, t and p-values, were also found within an acceptable range. The result also showed that, out of all the variables which were used, only three have a significant effect on the utilization rates of pedestrian zebra crossing while the remaining variables have an insignificant effect. The study concluded that among the different variables, Guardrail, number of lanes and Width of zebra crossing were the most influential variables. It is believed that the conclusions drawn from this research are expected to be useful to improve the state of pedestrian facilities in Malaysia.
  7. Ahmad J, Al Mamun A, Masukujjaman M, Mohamed Makhbul ZK, Mohd Ali KA
    Heliyon, 2023 Sep;9(9):e19134.
    PMID: 37681178 DOI: 10.1016/j.heliyon.2023.e19134
    This quantitative study investigates the relationships of green human resource management (GHRM) practices (e.g., green training and involvement, green recruitment, green performance management and compensation, and green transformational leadership) on green organizational culture and pro-environmental behavior, and the moderating role of green social capital and green values. This study adopts a cross-sectional design and collects quantitative data from 232 respondents working in top-to middle-level managerial positions in medium and large enterprises using a questionnaire survey after obtaining a list of companies from the Securities and Exchange Commission of Lahore, Rawalpindi, and Multan in Pakistan, applying the snowball sampling technique. A combined approach including partial least squares structural equation modeling and necessary condition analysis is employed to unravel the underlying mechanism between GHRM practices, green organizational culture, and pro-environmental behavior using Smart PLS version 4. The findings reveal that green transformational leadership (β = 0.267, p 
  8. Chan J, Craddock N, Swami V
    Body Image, 2023 Sep;46:1-8.
    PMID: 37149923 DOI: 10.1016/j.bodyim.2023.04.010
    Research on body image and racism has largely focused on how individual experiences of racism are associated with detrimental body image outcomes. However, research has not yet examined how resistance and empowerment against racism (REAR) - a repertoire of proactive strategies aimed at preventing or challenging racism both at the individual and collective levels - shape positive body image outcomes. Here, 236 women and 233 men who identified as belonging to racialised minority groups in the United Kingdom completed the REAR Scale - which measures REAR along four dimensions - as well as measures of body appreciation and body acceptance by others. Correlational analyses indicated significant inter-correlations between nearly all REAR domains and body image-related variables in men, whereas relationships in women were largely non-significant. Linear model analyses indicated that greater leadership for resistance against racism was significantly associated with higher body appreciation in women and men. Greater interpersonal confrontation of racism was significantly associated with both body appreciation and body acceptance by others in men, but not in women. These results suggest that REAR may play a role in shaping body image-related outcomes in people of colour, but that these effects are also be shaped by the intersection of gender and race.
    MeSH terms: Adult; Female; Gender Identity; Great Britain; Humans; Male; Minority Groups; Racism*
  9. Tai L, Saffery NS, Chin SP, Cheong SK
    Regen Med, 2023 Nov;18(11):839-856.
    PMID: 37671699 DOI: 10.2217/rme-2023-0085
    Aim: To profile and study the proteins responsible for the beneficial effect of the TNF-α-induced human umbilical cord mesenchymal stem cells (hUCMSCs) secretome in wound healing. Methods: The hUCMSCs secretome was generated with (induced) or without (uninduced) TNF-α and was subsequently analyzed by liquid chromatography-mass spectrometry, immunoassay and in vitro scratch assay. Results: Proteomic analysis revealed approximately 260 proteins, including 51 and 55 unique proteins in the induced and uninduced secretomes, respectively. Gene ontology analysis disclosed that differential proteins in the induced secretome mainly involved inflammation-related terms. The induced secretome, consisting of higher levels of FGFb, VEGF, PDGF and IL-6, significantly accelerated wound closure and enhanced MMP-13 secretion in HaCaT keratinocytes. Conclusion: The secretome from induced hUCMSCs includes factors that promote wound closure.
    MeSH terms: Humans; Umbilical Cord/metabolism; Wound Healing; Proteomics
  10. Zhao W, Irfan M
    Environ Sci Pollut Res Int, 2023 Oct;30(46):102772-102789.
    PMID: 37672158 DOI: 10.1007/s11356-023-29554-x
    In the face of the challenge of balancing urban economic development and environmental protection, the concept of a healthy city has emerged as a promising model for sustainable urban development. This study empirically investigates the impact of healthy city construction on green growth by utilizing a difference-in-difference model estimation on a panel dataset of 279 Chinese prefecture-level cities from 2007 to 2019. The findings reveal that healthy city construction significantly contributes to green growth, particularly in pilot cities, and this effect is observed across cities of different sizes and economic bases. Additionally, we identify two channels through which healthy city construction promotes green growth: enhancing innovation capacity and enriching human resources. These findings have implications not only for Chinese cities navigating the path towards green growth but also for other developing nations striving for economic transformation and environmentally sustainable development.
  11. El-Nouby MAM, Lim LW
    Anal Sci, 2023 Dec;39(12):2019-2029.
    PMID: 37672170 DOI: 10.1007/s44211-023-00416-3
    This article describes the fabrication of porous nicotinic acid-functionalized chito-oligosaccharide-bonded titania/silica hybrid monoliths (TiO2/SiO2@ChO-N) through a co-gelation sol-gel process. A capillary monolith with a well-defined and homogeneous structure was obtained by controlling the hydrolysis speed of titanium alkoxides in a sol mixture by using glycerol and acetylacetone. As a result of the functionalization with chito-oligosaccharides (ChO)-modified nicotinic acid, the obtained stationary phase provides superior physiochemical properties, such as a cationic hydrophilic surface, porosity, and mechanical strength. Scanning electron microscope and attenuated total reflectance-infrared spectroscopy were used to characterize the functionalized monolithic columns. The produced capillary columns showed high chromatographic performance with acceptable selectivity for charged analytes as well as organic polar compounds such as nucleic bases, nucleosides, carbamate pesticides, and strobilurin fungicides. The obtained results also indicated that the functionalized ChO's amino, amide, hydroxyl, and pyridinium ring moieties served as hydrophilic electrostatic traps for charged substances, in addition to stroing π-π interaction with the carbamate pesticides and strobilurin fungicides analytes via hydrogen bonding.
  12. Au SX, Mohd Padzil A, Muhd Noor ND, Matsumura H, Raja Abdul Rahman RNZ, Normi YM
    PLoS One, 2023;18(9):e0291012.
    PMID: 37672512 DOI: 10.1371/journal.pone.0291012
    BLEG-1 from Bacillus lehensis G1 is an evolutionary divergent B3 metallo-β-lactamase (MBL) that exhibited both β-lactamase and glyoxalase II (GLXII) activities. Sequence, phylogeny, biochemical and structural relatedness of BLEG-1 to B3 MBL and GLXII suggested BLEG-1 might be an intermediate in the evolutionary path of B3 MBL from GLXII. The unique active site cavity of BLEG-1 that recognizes both β-lactam antibiotics and S-D-lactoylglutathione (SLG) had been postulated as the key factor for its dual activity. In this study, dynamic ensembles of BLEG-1 and its substrate complexes divulged conformational plasticity and binding modes of structurally distinct substrates to the enzyme, providing better insights into its structure-to-function relationship and enzymatic promiscuity. Our results highlight the flexible nature of the active site pocket of BLEG-1, which is governed by concerted loop motions involving loop7+α3+loop8 and loop12 around the catalytic core, thereby moulding the binding pocket and facilitate interactions of BLEG-1 with both ampicillin and SLG. The distribution of (i) predominantly hydrophobic amino acids in the N-terminal domain, and (ii) flexible amino acids with polar and/or charged side chains in both N- and C-termini provide additional advantages to BLEG-1 in confining the aromatic group of ampicillin, and polar groups of SLG, respectively. The importance of these residues for substrates binding was further confirmed by the reduction in MBL and GLXII activities upon alanine substitutions of Ile-10, Phe-57, Arg-94, Leu-95, and Arg-159. Based on molecular dynamics simulation, mutational, and biochemical data presented herein, the catalytic mechanisms of BLEG-1 toward the hydrolysis of β-lactams and SLG were proposed.
    MeSH terms: Alanine*; Amino Acids; Ampicillin; Antifibrinolytic Agents*
  13. Yap FC, Wong WL, Chong VC, Bong CW, Lim LS
    Arthropod Struct Dev, 2023 Sep;76:101298.
    PMID: 37672818 DOI: 10.1016/j.asd.2023.101298
    The advancements in microscopic techniques have stimulated great interest in the muscular and neural architectures of invertebrates, specifically using muscle and neural structures to infer phylogenetic relationships. Here, we provide the data on the development of the muscular and nervous systems during the larval development of stalked barnacle, Octolasmis angulata using the phalloidin F-actin and immunohistochemical labelling (e.g. acetylated α-tubulin and serotonin) and confocal laser scanning microscopy analysis. All naupliar stages shared the same muscle and neural architectures with only the discrepancy in size. The nauplii have a complex muscle arrangement in their feeding apparatus and naupliar appendages. Most naupliar muscles undergo histolyse during the cyprid metamorphosis. The cyprid muscles form beneath the head shield at the end of nauplius VI. The naupliar and cyprid central nervous systems exhibit the typical tripartite brain comprising the protocerebrum, deutocerebrum and tritocerebrum. The serotonin-like immunoreactivity is mainly found in the naupliar brain, mandibular ganglia, cyprid brain and posterior ganglia. Our study revealed that numerous muscle and neural architectures in the naupliar and cyprids have phylogenetic significance, but future studies on the myoanatomy and neuroanatomy of other barnacle species are necessary to determine the homology of these structures.
    MeSH terms: Animals; Thoracica*; Central Nervous System; Muscles; Phylogeny; Serotonin
  14. Tan K, Waiho K, Tan K, Qiao Y, Lim LS, Yang X, et al.
    Biochem Biophys Res Commun, 2023 Oct 30;679:66-74.
    PMID: 37673004 DOI: 10.1016/j.bbrc.2023.08.066
    Vitellogenin (Vtg) serves as the precursor of yolk protein and exhibits widespread distribution in tissues, including in the ovary of both vertebrates and invertebrates. Vtg plays a critical role in facilitating oocyte maturation and embryonic development following oviposition. In this study, we have successfully elucidated the complete transcript sequence of TtVtg6-like from an ancient chelicerate Tachypleus tridentatus. The TtVtg6-like transcript encompassed a length of 4887 bp and encoded 1629 amino acids residues. Notably, TtVtg6-like was found to contain 25 exons. Furthermore, the molecular weight and isoelectric point of TtVtg6-like were determined to be 191.6 KDa and 6.73, respectively. Subsequent mRNA expression analysis demonstrated the specific expression of TtVtg6-like in ovary and yellow connective tissue. In addition, TtVtg6-like was located and distributed in both ovary and yellow connective tissue. Intriguingly, employing an siRNA approach to silence TtVtg6-like resulted in a decrease in TtVtg6-like transcription levels. Concomitantly, TtVtg6-like silencing led to increase production of ROS, ultimately resulting in DNA damage and cell apoptosis within the ovarian primary cell. The induction of apoptosis ovarian primary cells due to TtVtg6-like silencing was further corroborated through TUNEL assay and flow cytometry analysis. Overall, our findings underscore the significance of TtVtg6-like in ovarian cell development, revealing its potential association with ovarian cell apoptosis. Consequently, the insights gained from this study contribute to the future exploration of vitellogenesis and ovarian development in T. tridentatus.
  15. Kang J, Peng R, Feng J, Wei J, Li Z, Huang F, et al.
    BMJ Open, 2023 Sep 06;13(9):e075030.
    PMID: 37673450 DOI: 10.1136/bmjopen-2023-075030
    OBJECTIVE: To evaluate the health systems efficiency in China and Association of Southeast Asian Nations (ASEAN) countries from 2015 to 2020.

    DESIGN: Health efficiency analysis using data envelopment analysis (DEA) and stochastic frontier approach analysis.

    SETTING: Health systems in China and ASEAN countries.

    METHODS: DEA-Malmquist model and SFA model were used to analyse the health system efficiency among China and ASEAN countries, and the Tobit regression model was employed to analyse the factors affecting the efficiency of health system among these countries.

    RESULTS: In 2020, the average technical efficiency, pure technical efficiency and scale efficiency of China and 10 ASEAN countries' health systems were 0.700, 1 and 0.701, respectively. The average total factor productivity (TFP) index of the health systems in 11 countries from 2015 to 2020 was 0.962, with a decrease of 1.4%, among which the average technical efficiency index was 1.016, and the average technical progress efficiency index was 0.947. In the past 6 years, the TFP index of the health system in Malaysia was higher than 1, while the TFP index of other countries was lower than 1. The cost efficiency among China and ASEAN countries was relatively high and stable. The per capita gross domestic product (current US$) and the urban population have significant effects on the efficiency of health systems.

    CONCLUSIONS: Health systems inefficiency is existing in China and the majority ASEAN countries. However, the lower/middle-income countries outperformed high-income countries. Technical efficiency is the key to improve the TFP of health systems. It is suggested that China and ASEAN countries should enhance scale efficiency, accelerate technological progress and strengthen regional health cooperation according to their respective situations.

    MeSH terms: Asia; China; Government Programs*; Health Systems Agencies*; Gross Domestic Product
  16. Han HL, Nurcahyanto DA, Muhammad N, Lee YJ, Nguyen TTH, Kim SG, et al.
    Sci Rep, 2023 Sep 06;13(1):14684.
    PMID: 37673882 DOI: 10.1038/s41598-023-35108-5
    In the effort of isolating novel microbial species, the strain PL0132T was isolated from a fallen leaf under fresh water at a stream, which glided when grown on a tap water medium (without nutrients). The strain was determined to be Gram-negative, strictly aerobic, and rod-shaped, which grew optimally at 25 °C, pH 6-7, and the strain tolerates 1% (w/v) NaCl concentration. The complete genome of strain PL0132T comprises one contig with a sequencing depth of 76×, consisting of 8,853,064 base pairs and the genomic DNA G + C content was 46.7% (genome). 16S rRNA gene sequence analysis revealed that strain PL0132T represents a member of the phylum Bacteroidetes and is affiliated with the genus Spirosoma. Based on genomic, phenotypic, and chemotaxonomic characteristics, the strain PL0132T represents a novel species of the genus Spirosoma, for which the name Spirosoma foliorum sp. nov. is proposed (= KCTC 72228 T = InaCC B1447T).
    MeSH terms: RNA, Ribosomal, 16S/genetics; Plant Leaves; Acer*; Bacteroidetes; Cytophagaceae*
  17. Abd El-Aal AAA, Jayakumar FA, Lahiri C, Tan KO, Reginald K
    Sci Rep, 2023 Sep 06;13(1):14673.
    PMID: 37673929 DOI: 10.1038/s41598-023-41581-9
    Cryptides are a subfamily of bioactive peptides that exist in all living organisms. They are latently encrypted in their parent sequences and exhibit a wide range of biological activities when decrypted via in vivo or in vitro proteases. Cationic cryptides tend to be drawn to the negatively charged membranes of microbial and cancer cells, causing cell death through various mechanisms. This makes them promising candidates for alternative antimicrobial and anti-cancer therapies, as their mechanism of action is independent of gene mutations. In the current study, we employed an in silico approach to identify novel cationic cryptides with potential antimicrobial and anti-cancer activities in atypical and systematic strategy by reanalysis of a publicly available RNA-seq dataset of Pacific white shrimp (Penaus vannamei) in response to bacterial infection. Out of 12 cryptides identified, five were selected based on their net charges and potential for cell penetration. Following chemical synthesis, the cryptides were assayed in vitro to test for their biological activities. All five cryptides demonstrated a wide range of selective activity against the tested microbial and cancer cells, their anti-biofilm activities against mature biofilms, and their ability to interact with Gram-positive and negative bacterial membranes. Our research provides a framework for a comprehensive analysis of transcriptomes in various organisms to uncover novel bioactive cationic cryptides. This represents a significant step forward in combating the crisis of multi-drug-resistant microbial and cancer cells, as these cryptides neither induce mutations nor are influenced by mutations in the cells they target.
    MeSH terms: Animals; Biological Assay; Cations; Biofilms; Penaeidae*
  18. Rajan R, Al Jarallah M, Al-Zakwani I, Dashti R, Sulaiman K, Panduranga P, et al.
    Oman Med J, 2023 Jul;38(4):e529.
    PMID: 37674520 DOI: 10.5001/omj.2023.89
    OBJECTIVES: The Rajan's heart failure (R-hf) score was proposed to aid risk stratification in heart failure patients. The aim of this study was to validate R-hf risk score in patients with acute decompensated heart failure.

    METHODS: R-hf risk score is derived from the product estimated glomerular filtration rate (mL/min), left ventricular ejection fraction (%), and hemoglobin levels (g/dL) divided by N-terminal pro-brain natriuretic peptide (pg/mL). This was a multinational, multicenter, prospective registry of heart failure from seven countries in the Middle East. Univariable and multivariable logistic regression was applied.

    RESULTS: A total of 776 patients (mean age = 62.0±14.0 years, 62.4% males; mean left ventricular ejection fraction = 33.0±14.0%) were included. Of these, 459 (59.1%) presented with acute decompensated chronic heart failure. The R-hf risk score group (≤ 5) was marginally associated with a higher risk of all-cause cumulative mortality at three months (adjusted odds ratio (aOR) = 4.28; 95% CI: 0.90-20.30; p =0.067) and significantly at 12 months (aOR = 3.84; 95% CI: 1.23-12.00; p =0.021) when compared to those with the highest R score group (≥ 50).

    CONCLUSIONS: Lower R-hf risk scores are associated with increased risk of all-cause cumulative mortality at three and 12 months.

  19. Deng H, Zhang R, Wang C, Zhang B, Wang J, Wang S, et al.
    Front Psychiatry, 2023;14:1215963.
    PMID: 37674551 DOI: 10.3389/fpsyt.2023.1215963
    BACKGROUND: This multicenter, three-armed, parallel, single-blind randomized controlled trial (RCT) primarily aims to compare the efficacy of virtual reality exposure therapy (VRET) with that of acceptance and commitment therapy (ACT) and treatment as usual (TAU) to depreciate the degree of alcohol craving among alcohol use disorder patients who have undergone in-patient detoxification across four timelines (t0 = baseline prior to intervention, t1 = 4 weeks after baseline, t2 = 12 weeks after baseline, and t3 = 24 weeks after baseline). The secondary aims of this RCT are to compare the efficacy of VRET with that of ACT and TAU to alleviate the severity of alcohol use disorder, dissipate comorbid depressive and anxiety symptoms, and normalize event-related potential (ERP) in electroencephalogram (EEG) monitoring across the four timelines.

    METHODS: Initially, after 2 weeks of in-patient detoxification, 120 patients with alcohol use disorder will be randomized into three groups (VRET, ACT, and TAU control groups) via stratified permuted block randomization in a 1:1:1 ratio. Baseline assessment (t0) commences, whereby all the participants will be administered with sociodemographic, clinical, and alcohol use characteristics questionnaire, such as Alcohol Use Disorder Identification Test (AUDIT), Penn Alcohol Craving Scale (PACS), Hamilton Anxiety Rating Scale (HAM-A), and Hamilton Depression Rating Scale (HAM-D), while event-related potential (ERP) detection in electroencephalogram (EEG) will also be carried out. Then, 4 weeks of VRET, ACT, and non-therapeutic supportive activities will be conducted in the three respective groups. For the subsequent three assessment timelines (t1, t2, and t3), the alcohol use characteristic questionnaire, such as AUDIT, PACS, HAM-D, HAM-A, and ERP monitoring, will be re-administered to all participants.

    DISCUSSION: As data on the effects of non-pharmacological interventions, such as VRET and ACT, on the treatment of alcohol craving and preventing relapse in alcohol use disorder are lacking, this RCT fills the research gap by providing these important data to treating clinicians. If proven efficacious, the efficacy of VRET and ACT for the treatment of other substance use disorders should also be investigated in future.

    CLINICAL TRIAL REGISTRATION: NCT05841823 (ClinicalTrials.gov).

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