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Integration of MALDI glycotyping and NMR analysis to uncover an O-antigen substructure from pathogenic Escherichia coli O111

Lee JC, Urakami S, Hinou H

Int J Biol Macromol, 2024 Nov 04.
PMID: 39505162 DOI: 10.1016/j.ijbiomac.2024.137178

Escherichia coli O111 serotype is a critical pathogenic E. coli strain that causes severe, potentially fatal complications. Despite its reported variation, only one structure of the O-antigen polysaccharide from E. coli O111 has been reported. Here, a substructure of the O-antigen from E. coli O111 was characterized using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry and NMR analysis. MALDI glycotyping revealed differing O-antigen repeating unit masses of Δm/z 787 and 828 in the E. coli strains and lipopolysaccharides from the O111 serogroup. This variation was caused by the replacement of the hexose residue with hexosamine in the repeating units, which was further confirmed by LIFT-TOF/TOF analysis. Structural elucidation of the O111 substructure by NMR analysis further demonstrated replacement of the hydroxyl group with an N-acetyl group on the terminal glucose residue of the O-antigen pentasaccharide repeating unit. To our knowledge, this study is the first to provide a detailed structural analysis of a new O-antigen substructure from the E. coli O111 serogroup.
Fulltext Isolation of Industrial Important Bioactive Compounds from Microalgae

Balasubramaniam V, Gunasegavan RD, Mustar S, Lee JC, Mohd Noh MF

Molecules, 2021 Feb 10;26(4).
PMID: 33579001 DOI: 10.3390/molecules26040943

Microalgae are known as a rich source of bioactive compounds which exhibit different biological activities. Increased demand for sustainable biomass for production of important bioactive components with various potential especially therapeutic applications has resulted in noticeable interest in algae. Utilisation of microalgae in multiple scopes has been growing in various industries ranging from harnessing renewable energy to exploitation of high-value products. The focuses of this review are on production and the use of value-added components obtained from microalgae with current and potential application in the pharmaceutical, nutraceutical, cosmeceutical, energy and agri-food industries, as well as for bioremediation. Moreover, this work discusses the advantage, potential new beneficial strains, applications, limitations, research gaps and future prospect of microalgae in industry.
Fulltext Implementation of Food Safety Management Systems along with Other Management Tools (HAZOP, FMEA, Ishikawa, Pareto). The Case Study of Listeria monocytogenes and Correlation with Microbiological Criteria

Lee JC, Daraba A, Voidarou C, Rozos G, Enshasy HAE, Varzakas T

Foods, 2021 Sep 13;10(9).
PMID: 34574279 DOI: 10.3390/foods10092169

The food industry's failure in planning and designing of and in implementing a Food Safety Management System and its foundation elements leads, in most instances, to compromised food safety and subsequent foodborne illness outbreaks. This phenomenon was noticed, worldwide, for all food processors, but with a much higher incidence in the medium- and small-sized food processing plants. Our study focuses on the importance of Food Safety Management System (FSMS), Critical Control Points Hazard Analysis (HACCP) and the Prerequisite Programs (PRPs) as the foundation of HACCP, in preventing foodborne outbreaks. For emphasis, we make use of the example of organizational food safety culture failures and the lack of managerial engagement which resulted in a multi-state listeriosis outbreak in USA. Moreover, we correlate this with microbiological criteria. Implementation of food safety management systems (ISO 22000:2018) along with incorporation of management tools such as HAZOP, FMEA, Ishikawa and Pareto have proved to be proactive in the maintenance of a positive food safety culture and prevention of cross-contamination and fraud.
Fulltext Carica papaya L. Leaf: A Systematic Scoping Review on Biological Safety and Herb-Drug Interactions

Lim XY, Chan JSW, Japri N, Lee JC, Tan TYC

Evid Based Complement Alternat Med, 2021;2021:5511221.
PMID: 34040647 DOI: 10.1155/2021/5511221

INTRODUCTION: The Carica papaya L. leaf is gaining interest as a potential therapeutic agent for alleviating dengue- and non-dengue-associated thrombocytopaenia. In that regard, safety considerations are as important as efficacy potential. The safety evaluation of botanical products for human use is complicated by variable formulations, complex phytochemical composition, and extrinsic toxicants. This review aimed to systematically collate related safety clinical and preclinical data, as well as reports on herb-drug interactions of C. papaya leaf consumption.
METHODS: A systematic search using predetermined keywords on electronic databases (MEDLINE, Cochrane Library Central, LILACS, and Web of Science) and grey literature was conducted. Relevant clinical and preclinical studies were identified, screened, and analysed to present an overall safety profile of C. papaya leaf consumption.
RESULTS: A total of 41 articles were included (23 clinical, 5 ongoing trials, and 13 preclinical) for descriptive analysis on study characteristics, adverse reactions, toxicity findings, and herb-drug interactions, from which 13 randomised controlled and quasiexperimental trials were further assessed for risk of bias and reporting quality. Overall, C. papaya leaf consumption (in the form of juice and standardised aqueous extract) was well tolerated by adult humans for short durations (
Mechanical properties, permeability and microstructural characterisation of rice husk ash sustainable concrete with the addition of carbon nanotubes

Jing Y, Lee JC, Moon WC, Ng JL, Yew MK, Chu MY

Heliyon, 2024 Jun 30;10(12):e32780.
PMID: 39022029 DOI: 10.1016/j.heliyon.2024.e32780

This study investigated the effects of incorporating carbon nanotubes (CNTs) into rice husk ash (RHA) sustainable concrete on its mechanical properties, permeability and microstructure characterisation. Mechanical test results suggested that the addition of 0.10 % multiwalled CNTs (MWCNTs) yielded optimal results, with increases in the compressive strength, splitting tensile strength, flexural strength, and elastic modulus of the RHA concrete at 28 days of 7 %, 23.81 %, 17.5 %, and 1.0 %, respectively. However, with further addition of MWCNTs, the mechanical properties ultimately deteriorated. Further, the incorporation of CNTs enhanced the long-term performance of RHA sustainable concrete. The addition of 0.1 % MWCNTs and 15 % RHA yielded a 20 %, 14 %, and 66 % decrease in water absorption, porosity, and chloride diffusion coefficient compared to the mixture solely containing 15 % RHA. Scanning electron microscopy of this mixture revealed the filling and bridging effects of MWCNTs between the hydration products have enhanced the performance of RHA sustainable concrete.
Fulltext Experimental study on the shear behaviour of high-strength lightweight concrete beams incorporating graphene oxide

Hong X, Lee JC, Qian B, Yu M, Li Q, You P

Sci Rep, 2024 Nov 15;14(1):28199.
PMID: 39548244 DOI: 10.1038/s41598-024-79819-9

Graphene oxide (GO) has achieved significant progress in the material behaviour of cement-based materials. However, research on its structural behaviour in high-strength lightweight concrete (HSLWC) structures is limited, which restricts its engineering applications. This study focused on investigating the effects of low contents of GO on the shear behaviour of HSLWC beams. A total of four HSLWC beams with GO contents of 0, 0.01, 0.03 and 0.05% (by weight of cement) were designed to observe failure modes, load‒deformation curves, shear capacities, crack behaviour and load‒strain curves under four-point loading by a 300 kN servo loading device. The results revealed that all the beams exhibited shear compression failure. GO improved the shear capacity of the HSLWC beams, and this strengthening effect increased with increase in GO content. When the content of GO was 0.05%, the ultimate load of the beam reached a maximum, which was 39.2% greater than that of the control beam. GO can endow the HSLWC beams with a certain degree of ductility. In addition, a modified JGJ 12-2006 model was proposed to predict the shear capacity of HSLWC beams containing different GO contents on the basis of a comparison of typical models. This study can provide exploratory engineering practice for evaluating and designing GO-reinforced HSLWC structures.
Trigonelline attenuates the adipocyte differentiation and lipid accumulation in 3T3-L1 cells

Ilavenil S, Arasu MV, Lee JC, Kim DH, Roh SG, Park HS, et al.

Phytomedicine, 2014 Apr 15;21(5):758-65.
PMID: 24369814 DOI: 10.1016/j.phymed.2013.11.007

Trigonelline is a natural alkaloid mainly found in Trigonella Foenum Graecum (fenugreek) Fabaceae and other edible plants with a variety of medicinal applications. Therefore, we investigated the molecular mechanism of trigonelline (TG) on the inhibition of adipocyte differentiation and lipid accumulation in 3T3-L1 cells. Trigonelline suppressed lipid droplet accumulation in a concentration (75 and 100 μM) dependent manner. Treatment of adipocyte with of TG down regulates the peroxisome proliferator-activated receptor (PPARγ) and CCAAT element binding protein (C/EBP-α) mRNA expression, which leads to further down regulation of other gene such as adiponectin, adipogenin, leptin, resistin and adipocyte fatty acid binding protein (aP2) as compared with respective control cells on 5th and 10th day of differentiation. Further, addition of triognelline along with troglitazone to the adipocyte attenuated the troglitazone effects on PPARγ mediated differentiation and lipid accumulation in 3T3-L1 cells. Trigonelline might compete against troglitazone for its binding to the PPARγ. In addition, adipocyte treated with trigonelline and isoproterenol separately. Isoproterenol, a lipolytic agent which inhibits the fatty acid synthase and GLUT-4 transporter expression via cAMP mediated pathway, we found that similar magnitude response of fatty acid synthase and GLUT-4 transporter expression in trigonelline treated adipocyte. These results suggest that the trigonelline inhibits the adipogenesis by its influences on the expression PPARγ, which leads to subsequent down regulation of PPAR-γ mediated pathway during adipogenesis. Our findings provide key approach to the mechanism underlying the anti-adipogenic activity of trigonelline.
Fulltext Mirror, mirror on the wall, which phytochemicals in Clinacanthus nutans inhibits advanced glycation end products of them all?

Afzan A, Lee JC, Adam Z, Mustafa Khalid N, Gunasegavan RDN, Md Noh MF, et al.

Fitoterapia, 2024 Apr 09;175:105958.
PMID: 38604262 DOI: 10.1016/j.fitote.2024.105958

In our quest to discover advanced glycation end products (AGEs) inhibitors from Clinacanthus nutans (Burm.f.) Lindau leaves, we conducted a bioactivity-based molecular networking. This approach integrates LC-MS2 profiling and in vitro antiglycation data to predict bioactive compounds. We began by screening three extracts: 100% ethanol, 70% ethanol and 100% water alongside the in vitro antioxidant activity, total phenolics content (TPC) and schaftoside content. Among these extracts, 100% ethanol extract exhibited the highest total AGEs inhibition effects (IC50 = 80.18 ± 11.6 μg/mL), DPPH scavenging activity (IC50 = 747.40 ± 10.30 μg/mL) and TPC (26.54 ± 2.09 μg GAE /mg extract). Intriguingly, 100% ethanol extract contained the lowest amount of schaftoside, suggesting the involvement of other phytochemicals in the antiglycation effects. The molecular networking and in silico structural annotations of 401 LC-MS features detected in the fractions from 100% ethanol extract predicted 21 bioactive compounds (p 0.90), including several C40 carotenoids, alkaloids containing tetrapyrrole structures and fatty acids. On the contrary, all phenolics showed weak correlations with antiglycation effects. These predictions were further validated in vitro, where carotenoid lutein showed half maximal inhibitory concentration, IC50 = 96 ± 8 μM and selected flavonoid-C-glycosides exhibited weaker inhibitions (IC50 between 568 and 1922 μM). Notably, lutein content was higher in freeze-dried leaves (12.42 ± 0.82 mg/100 g) than oven-dried, although the former was associated with elevated mercury levels. In summary, C. nutans exhibited potential antiglycation and antioxidant activity, and lutein was identified as the main bioactive principle.
Developments in understanding and applying prebiotics in research and practice-an ISAPP conference paper

Scott KP, Grimaldi R, Cunningham M, Sarbini SR, Wijeyesekera A, Tang MLK, et al.

J Appl Microbiol, 2020 Apr;128(4):934-949.
PMID: 31446668 DOI: 10.1111/jam.14424

AIMS: The concept of using specific dietary components to selectively modulate the gut microbiota to confer a health benefit, defined as prebiotics, originated in 1995. In 2018, a group of scientists met at the International Scientific Association for Probiotics and Prebiotics annual meeting in Singapore to discuss advances in the prebiotic field, focussing on issues affecting functionality, research methodology and geographical differences.
METHODS AND RESULTS: The discussion ranged from examining scientific literature supporting the efficacy of established prebiotics, to the prospects for establishing health benefits associated with novel compounds, isolated from different sources.
CONCLUSIONS: While many promising candidate prebiotics from across the globe have been highlighted in preliminary research, there are a limited number with both demonstrated mechanism of action and defined health benefits as required to meet the prebiotic definition. Prebiotics are part of a food industry with increasing market sales, yet there are great disparities in regulations in different countries. Identification and commercialization of new prebiotics with unique health benefits means that regulation must improve and remain up-to-date so as not to risk stifling research with potential health benefits for humans and other animals.
SIGNIFICANCE AND IMPACT OF STUDY: This summary of the workshop discussions indicates potential avenues for expanding the range of prebiotic substrates, delivery methods to enhance health benefits for the end consumer and guidance to better elucidate their activities in human studies.
Fulltext Herbal-Based Formulation Containing Eurycoma longifolia and Labisia pumila Aqueous Extracts: Safe for Consumption?

Teh BP, Ahmad N, Ibnu Rasid EN, Zolkifli NA, Sastu Zakaria UR, Mohamed Yusoff N, et al.

Pharmaceuticals (Basel), 2021 Feb 10;14(2).
PMID: 33579048 DOI: 10.3390/ph14020142

A combined polyherbal formulation containing tongkat ali (Eurycoma longifolia) and kacip fatimah (Labisia pumila) aqueous extracts was evaluated for its safety aspect. A repeated dose 28-day toxicity study using Wistar rats was conducted where the polyherbal formulation was administered at doses 125, 500 and 2000 mg/kg body weight to male and female treatment groups daily via oral gavage, with rats receiving only water as the control group. In-life parameters measured include monitoring of food and water consumption and clinical and functional observations. On day 29, blood was collected for haematological and biochemical analysis. The rats were necropsied and the organs were collected for histopathological examination. This study showed that the combined formulation did not induce any significant toxicity effect at any dose level in terms of morbidity, mortality, behaviour, functional observation, body weight, food and water consumption, whole blood haematology and serum biochemistry. However, there were some microscopic changes in the histopathological examinations of some organs given 2000 mg/kg body weight, which may suggest an early response to the polyherbal formulation. From this study, the no observed adverse effect level is estimated to be more than 500 mg/kg body weight but not exceeding 2000 mg/kg body weight. The observed effects at the highest dose indicate the need for further study of longer dosing duration.
Efficacy of Fe-Mg-bimetallic biochar in stabilization of multiple heavy metals-contaminated soil and attenuation of toxicity in spinach (Spinacia oleracea L.)

Irshad MK, Lee JC, Aqeel M, Javed W, Noman A, Lam SS, et al.

Chemosphere, 2024 Aug 26;364:143184.
PMID: 39197684 DOI: 10.1016/j.chemosphere.2024.143184

Globally, soil contamination with heavy metals (HMs) pose serious threats to soil health, crop productivity, and human health. The present investigation involved synthesis and analysis of biochar with bimetallic combination of iron and magnesium (Fe-Mg-BC). Our study evaluated how Fe-Mg-BC affects the absorption of cadmium (Cd), lead (Pb), and copper (Cu) in spinach (Spinacia oleracea L.) and remediation of soil contaminated with multiple HMs. Results demonstrated the successful loading of iron (Fe) and magnesium (Mg) onto pristine biochar (BC) derived from peanut shells. The addition of Fe-Mg-BC (3%) notably increased spinach biomass, enhancing photosynthesis, transpiration, stomatal conductance, and intercellular CO2 levels by 22%, 21%, 103%, and 15.3%, respectively. Compared to control, Fe-Mg-BC (3%) suppressed metal-induced oxidative stress by boosting levels of superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) in roots by 40.9%, 57%, 54.8 %, and in shoots by 55.5%, 65.5%, and 37.4% in shoots, respectively. The Fe-Mg-BC effectively reduced the uptake of Cd, Pb, and Cu in spinach tissues by transforming their bioavailable fractions to non-bioavailable forms. The Fe-Mg-BC (3%) significantly reduced the mobility of Cd, Pb and Cu in soil and limited the concentration of Cd, Pb, and Cu in plant roots by 34.1%, 79.2%, 47%, and shoots by 56.3%, 43.3%, and 54.1%, respectively, compared to control. These findings underscore the potential of Fe-Mg-BC as a promising amendment for reclaiming soils contaminated with variety of HMs, thereby making a significant contribution to the promotion of safer food production.

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