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Discrimination of Malaysian stingless bee honey from different entomological origins based on physicochemical properties and volatile compound profiles using chemometrics and machine learning

Sharin SN, Sani MSA, Jaafar MA, Yuswan MH, Kassim NK, Manaf YN, et al.

Food Chem, 2021 Jun 01;346:128654.
PMID: 33461823 DOI: 10.1016/j.foodchem.2020.128654

Identification of honey origin based on specific chemical markers is important for honey authentication. This study is aimed to differentiate Malaysian stingless bee honey from different entomological origins (Heterotrigona bakeri, Geniotrigona thoracica and Tetrigona binghami) based on physicochemical properties (pH, moisture content, ash, total soluble solid and electrical conductivity) and volatile compound profiles. The discrimination pattern of 75 honey samples was observed using Principal Component Analysis (PCA), Hierarchical Clustering Analysis (HCA), Partial Least Square-Discriminant Analysis (PLS-DA), and Support Vector Machine (SVM). The profiles of H. bakeri and G. thoracica honey were close to each other, but clearly separated from T. binghami honey, consistent with their phylogenetic relationship. T. binghami honey is marked by significantly higher electrical conductivity, moisture and ash content, and high abundance of 2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde, 2,6,6-trimethyl-1-cyclohexene-1-acetaldehyde and ethyl 2-(5-methyl-5-vinyltetrahydrofuran-2-yl)propan-2-yl carbonate. Copaene was proposed as chemical marker for G. thoracica honey. The potential of different parameters that aid in honey authentication was highlighted.
Comparative database search engine analysis on massive tandem mass spectra of pork-based food products for halal proteomics

Amir SH, Yuswan MH, Aizat WM, Mansor MK, Desa MNM, Yusof YA, et al.

J Proteomics, 2021 06 15;241:104240.
PMID: 33894373 DOI: 10.1016/j.jprot.2021.104240

Mass spectrometry-based proteomics relies on dedicated software for peptide and protein identification. These software include open-source or commercial-based search engines; wherein, they employ different algorithms to establish their scoring and identified proteins. Although previous comparative studies have differentiated the proteomics results from different software, there are still yet studies specifically been conducted to compare and evaluate the search engine in the field of halal analysis. This is important because the halal analysis is often using commercial meat samples that have been subjected to various processing, further complicating its analysis. Thus, this study aimed to assess three open-source search engines (Comet, X! Tandem, and ProteinProspector) and a commercial-based search engine (ProteinPilot™) against 135 raw tandem mass spectrometry data files from 15 types of pork-based food products for halal analysis. Each database search engine contained high false-discovery rate (FDR); however, a post-searching algorithm called PeptideProphet managed to reduce the FDR, except for ProteinProspector and ProteinPilot™. From this study, the combined database search engine (executed by iProphet) reveals a thorough protein list for pork-based food products; wherein the most abundant proteins are myofibrillar proteins. Thus, this proteomics study will aid the identification of potential peptide and protein biomarkers for future precision halal analysis. SIGNIFICANCE: A critical challenge of halal proteomics is the availability of a database to confirm the inferential peptides as well as proteins. Currently, the established database such as UniProtKB is related to animal proteome; however, the halal proteomics is related to the highly processed meat-based food products. This study highlights the use of different database search engines (Comet, X! Tandem, ProteinProspector, and ProteinPilot™) and their respective algorithms to analyse 135 raw tandem mass spectrometry data files from 15 types of pork-based food products. This is the first attempt that has compared different database search engines in the context of halal proteomics to ensure the effectiveness of controlling the FDR. Previous studies were just focused on the advantages of a certain algorithm over another. Moreover, other previous studies also have mainly reported the use of mass spectrometry-based shotgun proteomics for meat authentication (the most similar field to halal analysis), but none of the studies were reported on halal aspects that used samples originated from highly processed food products. Hence, a systematic comparative study is duly needed for a more comprehensive and thorough proteomics analysis for such samples. In this study, our combinatorial approach for halal proteomics results from the different search engines used (Comet, X! Tandem, and ProteinProspector) has successfully generated a comprehensive spectral library for the pork-based meat products. This combined spectral library is freely available at https://data.mendeley.com/datasets/6dmm8659rm/3. Thus far, this is the first and new attempt at establishing a spectral library for halal proteomics. We also believe this study is a pioneer for halal proteomics that aimed at non-conventional and non-model organism proteomics, protein analytics, protein bioinformatics, and potential biomarker discovery.
Fulltext Antibacterial Activity and Mode of Action of β-caryophyllene on Bacillus cereus

Moo CL, Yang SK, Osman MA, Yuswan MH, Loh JY, Lim WM, et al.

Pol J Microbiol, 2020;69:1-6.
PMID: 32162852 DOI: 10.33073/pjm-2020-007

Natural products such as essential oils (EOs) are secondary metabolites that can be obtained from either plant or animal sources or produced by microorganisms. Much attention has been given to exploring the use of secondary metabolites as natural antibacterial agents. This study investigates the antibacterial activity and mechanism of β-caryophyllene, a compound that can be found in various EOs, against Bacillus cereus. The minimum inhibitory concentration of β-caryophyllene against B. cereus was 2.5% (v/v), whereas killing kinetics of β-caryophyllene at minimum inhibitory concentration recorded complete bactericidal activity within 2 hours. Zeta-potential measurement in the cells treated with half the minimum inhibitory concentration of β-caryophyllene at 1.25% (v/v) showed an increase in the membrane permeability surface charge to -3.98 mV, compared to untreated cells (-5.46 mV). Intracellular contents leakage of UV-absorbing materials was detected in the cells treated with β-caryophyllene. Additionally, β-caryophyllene does not interfere with the efflux activity of B. cereus via the ethidium bromide influx/efflux activity. The results revealed that β-caryophyllene was able to alter membrane permeability and integrity of B. cereus, leading to membrane damage and intracellular content leakage, which eventually caused cell death.
Natural products such as essential oils (EOs) are secondary metabolites that can be obtained from either plant or animal sources or produced by microorganisms. Much attention has been given to exploring the use of secondary metabolites as natural antibacterial agents. This study investigates the antibacterial activity and mechanism of β-caryophyllene, a compound that can be found in various EOs, against Bacillus cereus. The minimum inhibitory concentration of β-caryophyllene against B. cereus was 2.5% (v/v), whereas killing kinetics of β-caryophyllene at minimum inhibitory concentration recorded complete bactericidal activity within 2 hours. Zeta-potential measurement in the cells treated with half the minimum inhibitory concentration of β-caryophyllene at 1.25% (v/v) showed an increase in the membrane permeability surface charge to –3.98 mV, compared to untreated cells (–5.46 mV). Intracellular contents leakage of UV-absorbing materials was detected in the cells treated with β-caryophyllene. Additionally, β-caryophyllene does not interfere with the efflux activity of B. cereus via the ethidium bromide influx/efflux activity. The results revealed that β-caryophyllene was able to alter membrane permeability and integrity of B. cereus, leading to membrane damage and intracellular content leakage, which eventually caused cell death.
Fulltext Sodium lignosulfonate improves shoot growth of Oryza sativa via enhancement of photosynthetic activity and reduced accumulation of reactive oxygen species

Kok AD, Wan Abdullah WMAN, Tang CN, Low LY, Yuswan MH, Ong-Abdullah J, et al.

Sci Rep, 2021 06 24;11(1):13226.
PMID: 34168171 DOI: 10.1038/s41598-021-92401-x

Lignosulfonate (LS) is a by-product obtained during sulfite pulping process and is commonly used as a growth enhancer in plant growth. However, the underlying growth promoting mechanism of LS on shoot growth remains largely unknown. Hence, this study was undertaken to determine the potential application of eco-friendly ion-chelated LS complex [sodium LS (NaLS) and calcium LS (CaLS)] to enhance recalcitrant indica rice MR 219 shoot growth and to elucidate its underlying growth promoting mechanisms. In this study, the shoot apex of MR 219 rice was grown on Murashige and Skoog medium supplemented with different ion chelated LS complex (NaLS and CaLS) at 100, 200, 300 and 400 mg/L The NaLS was shown to be a better shoot growth enhancer as compared to CaLS, with optimum concentration of 300 mg/L. Subsequent comparative proteomic analysis revealed an increase of photosynthesis-related proteins [photosystem II (PSII) CP43 reaction center protein, photosystem I (PSI) iron-sulfur center, PSII CP47 reaction center protein, PSII protein D1], ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), carbohydrate metabolism-related proteins (glyceraldehyde-3-phosphate dehydrogenase 3, fructose-bisphosphate aldolase) and stress regulator proteins (peptide methionine sulfoxide reductase A4, delta-1-pyrroline-5-carboxylate synthase 1) abundance in NaLS-treated rice as compared to the control (MSO). Consistent with proteins detected, a significant increase in biochemical analyses involved in photosynthetic activities, carbohydrate metabolism and protein biosynthesis such as total chlorophyll, rubisco activity, total sugar and total protein contents were observed in NaLS-treated rice. This implies that NaLS plays a role in empowering photosynthesis activities that led to plant growth enhancement. In addition, the increased in abundance of stress regulator proteins were consistent with low levels of peroxidase activity, malondialdehyde content and phenylalanine ammonia lyase activity observed in NaLS-treated rice. These results suggest that NaLS plays a role in modulating cellular homeostasis to provide a conducive cellular environment for plant growth. Taken together, NaLS improved shoot growth of recalcitrant MR 219 rice by upregulation of photosynthetic activities and reduction of ROS accumulation leading to better plant growth.
Impact of harvesting seasons on physicochemical properties and volatile compound profiles of Malaysian stingless bee honey analysed using chemometrics and support vector machine

Sharin SN, Abdullah Sani MS, Kassim NK, Yuswan MH, Abd Aziz A, Jaafar MA, et al.

Food Chem, 2024 Jan 19;444:138429.
PMID: 38330597 DOI: 10.1016/j.foodchem.2024.138429

Stingless bee honey's nutritional value is gaining attention, but the impact of harvesting seasons, specifically the rainy (September 2018) and dry (February 2019) seasons in Malaysia on the honey's physicochemical properties and volatile compounds remains insufficiently explored. This research revealed marginal differences in the physicochemical properties between seasons. However, through individual bee species and cumulative data analysis, honey samples were effectively differentiated based on harvesting seasons. A set of seventeen volatile compounds were identified as potential chemical markers for distinguishing H. bakeri, G. thoracica, and T. binghami honey between rainy and dry seasons. For cumulative data, four significant markers were proposed. These discrimination methods and chemical markers can serve as valuable references in distinguishing stingless bee honey, whether its entomological origin is specified or not between rainy and dry seasons.
Hydroxyproline determination for initial detection of halal-critical food ingredients (gelatin and collagen)

Yuswan MH, A Jalil NH, Mohamad H, Keso S, Mohamad NA, Tengku Md Yusoff TS, et al.

Food Chem, 2021 Feb 01;337:127762.
PMID: 32777563 DOI: 10.1016/j.foodchem.2020.127762

Gelatin and collagen are considered halal-critical ingredients as they are typically derived from either bovine or porcine animals. Current analytical methods for determining the sources of gelatin and collagen suffer from limitations in terms of robustness and false positives in peptide matching. Thus, the aim of this study was to investigate the utility of monitoring hydroxyproline, a signature amino acid for gelatin and collagen, for identifying potentially haram foodstuffs. To determine the hydroxyproline profiles among animal- and plant-based samples, one-way univariate analysis of variance followed by pair-wise comparison was used to establish statistical significance. Multivariate chemometric analysis through principal component analysis revealed a discrete distribution pattern among 59 samples due to hydroxyproline variability. Finally, inter- and intra-laboratory comparisons demonstrated the validity and robustness of hydroxyproline determination according to ISO 17025. Thus, this preliminary identification technique will aid the identification of potentially haram foodstuffs.
Fulltext The characteristics and roles of antimicrobial peptides as potential treatment for antibiotic-resistant pathogens: a review

Zainal Baharin NH, Khairil Mokhtar NF, Mohd Desa MN, Gopalsamy B, Mohd Zaki NN, Yuswan MH, et al.

PeerJ, 2021;9:e12193.
PMID: 35003909 DOI: 10.7717/peerj.12193

The emergence of antibiotic-resistant bacteria has become a significant and ever-increasing threat to global public health, increasing both morbidity and mortality rates, and the financial burden on health services. Infection by drug-resistant bacteria is anticipated to contribute to the demise of almost 10 million people by the year 2050 unless a competent and effective response is devised to engage with this issue. The emergence and spread of resistance are commonly caused by the excessive or inappropriate use of antibiotics and substandard pharmaceuticals. It arises when pathogens adapt to different conditions and develop self-defence mechanisms. Currently, novel antimicrobial peptides (AMPs) have been reported to be the sole cure for some clinical cases of infectious diseases such as sepsis and skin infections, although these agents may, on occasion, require administration together with an adjunctive low-dose antibiotic. Although AMPs are a promising alternative form of anti-microbial therapy and easily applied in the medical sector, they still have limitations that should not be taken lightly. Hence, this review explores the characteristics, advantages and disadvantages of AMPs for their potential in treating antibiotic-resistant pathogens.