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Microencapsulated Lactobacillus plantarum LAB12 Showed No Sign of Acute or Sub-chronic Toxicity In Vivo

Fareez IM, Lim SM, Ramasamy K

Probiotics Antimicrob Proteins, 2019 06;11(2):447-459.
PMID: 30003409 DOI: 10.1007/s12602-018-9442-7

Lactic acid bacteria (LAB) with probiotic properties are useful options for prophylactic and therapeutic applications against gastrointestinal diseases. The safety of probiotics should, however, be verified before incorporation into food or drinks. The present study had encapsulated Lactobacillus plantarum LAB12 within microcapsules that could withstand extremely high temperature (up to 100 °C) during pelletisation. The microencapsulated LAB12 were then tested for their acute (single dosing) and sub-chronic (a 90-day feeding) toxicity. For acute toxicity study, six male Sprague-Dawley rats were being administered with a single dose of freeze-dried microencapsulated LAB12 at 11 log CFU/kg BW through oral gavage. No clear treatment-related effects were observed after 14 days. For sub-chronic toxicity study, rodents were randomly divided into four groups (6 rats/sex/group) and treated with 0, 8, 9 and 10 log CFU/kg BW of microencapsulated LAB12 in pellet form. No mortality or treatment-related findings were observed in terms of clinical body weight, water intake, or food consumption. No treatment-related adverse effects were observed in blood and tissue samples. The no-observed-adverse-effect-level (NOAEL) for microencapsulated LAB12 was 2.5 × 1010 CFU/kg BW for both genders. These results imply that LAB12 are likely non-pathogenic and non-toxic.
Chemoprevention by Microencapsulated Lactiplantibacillus Plantarum LAB12 Against Orthotopic Colorectal Cancer Mice is Associated with Apoptosis and Anti-angiogenesis

Fareez IM, Lim SM, Ramasamy K

Probiotics Antimicrob Proteins, 2024 Feb;16(1):99-112.
PMID: 36508139 DOI: 10.1007/s12602-022-10020-y

The pathogenesis of colorectal cancer (CRC) is associated with gut dysbiosis that is attributed to unhealthy lifestyles and dietary habits. Consumption of microencapsulated probiotics may potentially restore the gut microbiota in favour of prevention against CRC. This study determined the fate of microencapsulated Lactiplantibacillus plantarum (formerly known as Lactobacillus plantarum) LAB12 in the gastrointestinal tract (GIT) and assessed the chemopreventive effect of microencapsulated L. plantarum LAB12 in vivo. The targeted release of L. plantarum LAB12 from Alg-based microcapsules at the stomach, ileum, caecum and colon of Sprague-Dawley rats was examined by confocal microscopy and qPCR. Microcapsules loaded with L. plantarum LAB12 remained intact in the stomach. Free L. plantarum LAB12 were present in abundance (> 7 log CFU) only in the intestines. Subsequently, the chemopreventive properties of microencapsulated L. plantarum LAB12 were validated against NU/NU nude mice bearing orthotopic transplanted CT-26 CRC (12 female mice; 4-6 weeks old; 20-22 g; n = 6/group). Orthotopic mice pre-supplemented with microencapsulated L. plantarum LAB12 (10 log CFU kg-1 BW for 11 weeks) were presented with significantly (p
Chitosan coated alginate-xanthan gum bead enhanced pH and thermotolerance of Lactobacillus plantarum LAB12

Fareez IM, Lim SM, Mishra RK, Ramasamy K

Int J Biol Macromol, 2015 Jan;72:1419-28.
PMID: 25450046 DOI: 10.1016/j.ijbiomac.2014.10.054

The vulnerability of probiotics at low pH and high temperature has limited their optimal use as nutraceuticals. This study addressed these issues by adopting a physicochemical driven approach of incorporating Lactobacillus plantarum LAB12 into chitosan (Ch) coated alginate-xanthan gum (Alg-XG) beads. Characterisation of Alg-XG-Ch, which elicited little effect on bead size and polydispersity, demonstrated good miscibility with improved bead surface smoothness and L. plantarum LAB12 entrapment when compared to Alg, Alg-Ch and Alg-XG. Sequential incubation of Alg-XG-Ch in simulated gastric juice and intestinal fluid yielded high survival rate of L. plantarum LAB12 (95%) at pH 1.8 which in turn facilitated sufficient release of probiotics (>7 log CFU/g) at pH 6.8 in both time- and pH-dependent manner. Whilst minimising viability loss at 75 and 90 °C, Alg-XG-Ch improved storage durability of L. plantarum LAB12 at 4 °C. The present results implied the possible use of L. plantarum LAB12 incorporated in Alg-XG-Ch as new functional food ingredient with health claims.
Role of the CXCR4-SDF1-HMGB1 pathway in the directional migration of cells and regeneration of affected organs

Haque N, Fareez IM, Fong LF, Mandal C, Abu Kasim NH, Kacharaju KR, et al.

World J Stem Cells, 2020 Sep 26;12(9):938-951.
PMID: 33033556 DOI: 10.4252/wjsc.v12.i9.938

In recent years, several studies have reported positive outcomes of cell-based therapies despite insufficient engraftment of transplanted cells. These findings have created a huge interest in the regenerative potential of paracrine factors released from transplanted stem or progenitor cells. Interestingly, this notion has also led scientists to question the role of proteins in the secretome produced by cells, tissues or organisms under certain conditions or at a particular time of regenerative therapy. Further studies have revealed that the secretomes derived from different cell types contain paracrine factors that could help to prevent apoptosis and induce proliferation of cells residing within the tissues of affected organs. This could also facilitate the migration of immune, progenitor and stem cells within the body to the site of inflammation. Of these different paracrine factors present within the secretome, researchers have given proper consideration to stromal cell-derived factor-1 (SDF1) that plays a vital role in tissue-specific migration of the cells needed for regeneration. Recently researchers recognized that SDF1 could facilitate site-specific migration of cells by regulating SDF1-CXCR4 and/or HMGB1-SDF1-CXCR4 pathways which is vital for tissue regeneration. Hence in this study, we have attempted to describe the role of different types of cells within the body in facilitating regeneration while emphasizing the HMGB1-SDF1-CXCR4 pathway that orchestrates the migration of cells to the site where regeneration is needed.
Fulltext Color Stability of a New Rice Husk Composite in Comparison with Conventional Composites after Exposure to Commonly Consumed Beverages in Malaysia

Raja KK, Hari P, Chin MQK, Singbal K, Fareez IM

Int J Dent, 2019;2019:9753431.
PMID: 31191655 DOI: 10.1155/2019/9753431

Objective: To evaluate the color stability of a new organic rice husk nanocomposite as compared to four conventional composites after exposure to commonly consumed beverages in Malaysia.
Methods: One hundred and twenty-five disk samples were prepared from a new rice husk-based composite and four other conventional methacrylate-based light-cured composites of shade A2. The samples were immersed in four commonly consumed beverages: coco-based drink, kopi, Chinese tea, and teh tarik for four weeks. The color measurements were carried out every week using the reflectance spectrophotometer according to the CIE L
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color system. Color changes of samples (ΔE) in each week were calculated. Statistical analysis was carried out by performing a mixed ANOVA and Tukey's post hoc test in order to analyse the differences in ΔE.
Results: The findings revealed a statistically significant difference of ΔE reading (p < 0.05) among all composites immersed in all four beverages after four weeks. Rice husk composites exhibited lesser color stability as compared to Ceram.X One Universal (p < 0.001) and G-aenial Universal Flo (p < 0.001) but showed higher color stability compared to Solare-X (p < 0.001) and Neofil (p < 0.001). Coffee and Chinese tea had the most significant impact on color changes (p < 0.05) observed in all composites over four weeks of study.
Conclusion: Rice husk composite showed acceptable color stability. It can be considered as an alternative to conventional composites due to its eco-friendly properties.
Cellulose Derivatives Enhanced Stability of Alginate-Based Beads Loaded with Lactobacillus plantarum LAB12 against Low pH, High Temperature and Prolonged Storage

Fareez IM, Lim SM, Zulkefli NAA, Mishra RK, Ramasamy K

Probiotics Antimicrob Proteins, 2018 09;10(3):543-557.
PMID: 28493103 DOI: 10.1007/s12602-017-9284-8

The susceptibility of probiotics to low pH and high temperature has limited their use as nutraceuticals. In this study, enhanced protection of probiotics via microencapsulation was achieved. Lactobacillus plantarum LAB12 were immobilised within polymeric matrix comprised of alginate (Alg) with supplementation of cellulose derivatives (methylcellulose (MC), sodium carboxymethyl cellulose (NaCMC) or hydroxypropyl methylcellulose (HPMC)). L. plantarum LAB12 encapsulated in Alg-HPMC(1.0) and Alg-MC(1.0) elicited improved survivability (91%) in simulated gastric conditions and facilitated maximal release (∼100%) in simulated intestinal condition. Alg-HPMC(1.0) and Alg-MC(1.0) significantly reduced (P 7 log CFU g-1. Alg-MC and Alg-HPMC improved the survival of LAB12 against simulated gastric condition (9.24 and 9.55 log CFU g-1, respectively), temperature up to 90 °C (9.54 and 9.86 log CFU g-1, respectively) and 4-week of storage at 4 °C (8.61 and 9.23 log CFU g-1, respectively) with sustained release of probiotic in intestinal condition (>9 log CFU g-1). These findings strongly suggest the potential of cellulose derivatives supplemented Alg bead as protective micro-transport for probiotic strains. They can be safely incorporated into new functional food or nutraceutical products.
Molecular and Epigenetic Basis of Extracellular Vesicles Cell Repair Phenotypes in Targeted Organ-specific Regeneration

Fareez IM, Seng WY, Zaki RM, Shafiq A, Izwan IM

Curr Mol Med, 2022;22(2):132-150.
PMID: 33568034 DOI: 10.2174/1566524021666210210121905

Extracellular vesicles (EVs), which are released by most of the cells, constitute a new system of cell-cell communication by transporting DNA, RNA, and proteins in various vesicles namely exosomes, apoptotic bodies, protein complexes, high-density lipid (HDL) microvesicles, among others. To ensure accurate regulation of somatic stem cell activity, EVs function as an independent metabolic unit mediating the metabolic homeostasis and pathophysiological of several diseases such as cardiovascular diseases, metabolic diseases, neurodegenerative diseases, immune diseases, and cancer. Whist examining the EV biomolecules cargos and their microenvironments that lead to epigenetic alteration of the cell in tissue regeneration, studies have gained further insights into the biogenesis of EVs and their potential roles in cell biology and pathogenicity. Due to their small size, non-virulence, flexibility, and ability to cross biological barriers, EVs have promising therapeutic potentials in various diseases. In this review, we describe EV's mechanism of action in intercellular communication and transfer of biological information as well as some details about EVinduced epigenetic changes in recipient cells that cause phenotypic alteration during tissue regeneration. We also highlight some of the therapeutic potentials of EVs in organ-specific regeneration.
Application of Platelet-Rich Plasma as a Stem Cell Treatment - an Attempt to Clarify a Common Public Misconception

Fareez IM, Liew FF, Widera D, Mayeen NF, Mawya J, Abu Kasim NH, et al.

Curr Mol Med, 2024;24(6):689-701.
PMID: 37171013 DOI: 10.2174/1566524023666230511152646

In recent years, there has been a significant increase in the practice of regenerative medicine by health practitioners and direct-to-consumer businesses globally. Among different tools of regenerative medicine, platelet-rich plasma (PRP) and stem cell-based therapies have received considerable attention. The use of PRP, in particular, has gained popularity due to its easy access, simple processing techniques, and regenerative potential. However, it is important to address a common misconception amongst the general public equating to PRP and stem cells due to the demonstrated efficacy of PRP in treating musculoskeletal and dermatological disorders. Notably, PRP promotes regeneration by providing growth factors or other paracrine factors only. Therefore, it cannot replenish or replace the lost cells in conditions where a large number of cells are required to regenerate tissues and/or organs. In such cases, cellbased therapies are the preferred option. Additionally, other tools of regenerative medicine, such as bioprinting, organoids, and mechanobiology also rely on stem cells for their success. Hence, healthcare and commercial entities offering direct-to-customer regenerative therapies should not mislead the public by claiming that the application of PRP is a stem cell-based therapy. Furthermore, it is important for regulatory bodies to strictly monitor these profit-driven entities to prevent them from providing unregulated regenerative treatments and services that claim a broad variety of benefits with little proof of efficacy, safety concerns, and obscure scientific justification.
Fulltext Selective phytochemicals targeting pancreatic stellate cells as new anti-fibrotic agents for chronic pancreatitis and pancreatic cancer

Ramakrishnan P, Loh WM, Gopinath SCB, Bonam SR, Fareez IM, Mac Guad R, et al.

Acta Pharm Sin B, 2020 Mar;10(3):399-413.
PMID: 32140388 DOI: 10.1016/j.apsb.2019.11.008

Activated pancreatic stellate cells (PSCs) have been widely accepted as a key precursor of excessive pancreatic fibrosis, which is a crucial hallmark of chronic pancreatitis (CP) and its formidable associated disease, pancreatic cancer (PC). Hence, anti-fibrotic therapy has been identified as a novel therapeutic strategy for treating CP and PC by targeting PSCs. Most of the anti-fibrotic agents have been limited to phase I/II clinical trials involving vitamin analogs, which are abundant in medicinal plants and have proved to be promising for clinical application. The use of phytomedicines, as new anti-fibrotic agents, has been applied to a variety of complementary and alternative approaches. The aim of this review was to present a focused update on the selective new potential anti-fibrotic agents, including curcumin, resveratrol, rhein, emodin, green tea catechin derivatives, metformin, eruberin A, and ellagic acid, in combating PSC in CP and PC models. It aimed to describe the mechanism(s) of the phytochemicals used, either alone or in combination, and the associated molecular targets. Most of them were tested in PC models with similar mechanism of actions, and curcumin was tested intensively. Future research may explore the issues of bioavailability, drug design, and nano-formulation, in order to achieve successful clinical outcomes with promising activity and tolerability.
Molecular Regulatory Roles of Long Non-coding RNA HOTTIP: An Overview in Gastrointestinal Cancer

Hamid UZ, Sim MS, Guad RM, Subramaniyan V, Sekar M, Fuloria NK, et al.

Curr Mol Med, 2021 Aug 06.
PMID: 34365949 DOI: 10.2174/1566524021666210806162848

Gastrointestinal (GI) cancers presented an alarmingly high number of new cancer cases worldwide and highly characterised with poor prognosis. The poor overall survival is mainly due to late detection and emerging challenges in treatment, particularly chemoresistance. Thus, the identification of novel molecular targets in GI cancer is highly regarded as the main focus. Recently, long non-coding RNAs (lncRNAs) have been discovered as a potential novel molecular target for combating cancer, as it is highly associated with carcinogenesis and has a great impact on cancer progression. Amongst lncRNAs, HOTIIP has demonstrated a prominent oncogenic regulation in cancer progression, particularly in GI cancers, including oesophageal cancer, gastric cancer, hepatocellular carcinoma, pancreatic cancer and colorectal cancer. This review aimed to present a focused update on the regulatory roles of HOTTIP in GI cancer progression and chemoresistance, as well as deciphering the associated molecular mechanisms underlying their impact on cancer phenotypes and chemoresistance and the key molecules involved. It has been reported that it regulates the expression of various genes and proteins in GI cancers that impacts on the cellular functions, including proliferation, adhesion, migration and invasion, apoptosis, chemosensitivity and tumour differentiation. Furthermore, HOTTIP was also discovered to have a higher diagnostic value as compared to existing diagnostic biomarkers. In overall, HOTTIP has presented itself as a novel therapeutic target and potential diagnostic biomarker in the development of GI cancer treatment.
Human Exfoliated Deciduous Teeth Stem Cells: Features and Therapeutic Effects on Neurogenerative and Hepatobiliary-pancreatic Diseases

Wahab NWA, Guad RM, Subramaniyan V, Fareez IM, Choy KW, Bonam SR, et al.

Curr Stem Cell Res Ther, 2021;16(5):563-576.
PMID: 32957893 DOI: 10.2174/1574888X15999200918105623

Stem cells can multiply into more cells with similar types in an undifferentiated form and differentiate into other types of cells. The great success and key essence of stem cell technology is the isolation of high-quality Mesenchymal Stem Cells (MSCs) with high potency, either with multipotent or pluripotent property. In this line, Stem cells from Human Exfoliated Deciduous teeth (SHEDs) are highly proliferative stem cells from dental pulp and have multipoint differentiation capacity. These cells play a pivotal role in regenerative medicine, such as cell repair associated with neurodegenerative, hepatobiliary, and pancreatic diseases. In addition, stem cell therapy has been widely used to regulate immune response and repair of tissue lesions. This overview captured the differential biological characteristics, and the potential role of stem cell technology and paid special attention to human welfare SHEDs in eliminating the above-mentioned diseases. This review provides further insights into stem cell technology by expanding the therapeutic potential of SHEDs in tissue engineering and cell organ repairs.

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