A Conceptual Model Relationship between Industry 4.0-Food-Agriculture Nexus and Agroecosystem: A Literature Review and Knowledge Gaps.

With the expected colonization of human daily life by artificial intelligence, including in industry productivity, the deployment of Industry 4.0 (I4) in the food agriculture industry (FAI) is expected to revolutionize and galvanize food production to increase the efficiency of the industry's production and to match, in tandem, a country's gross domestic productivity. Based on a literature review, there have been almost no direct relationships between the I4-Food-Agriculture (I4FA) Nexus and the agroecosystem. This study aimed to evaluate the state-of-the-art relationships between the I4FA Nexus and the agroecosystem and to discuss the challenges in the sustainable FAI that can be assisted by the I4 technologies. This objective was fulfilled by (a) reviewing all the relevant publications and (b) drawing a conceptual relationship between the I4FA Nexus and the agroecosystem, in which the I4FA Nexus is categorized into socio-economic and environmental (SEE) perspectives. Four points are highlighted in the present review. First, I4 technology is projected to grow in the agricultural and food sectors today and in the future. Second, food agriculture output may benefit from I4 by considering the SEE benefits. Third, implementing I4 is a challenging journey for the sustainable FAI, especially for the small to medium enterprises (SMEs). Fourth, environmental, social, and governance (ESG) principles can help to manage I4's implementation in agriculture and food. The advantages of I4 deployment include (a) social benefits like increased occupational safety, workers' health, and food quality, security, and safety; (b) economic benefits, like using sensors to reduce agricultural food production costs, and the food supply chain; and (c) environmental benefits like reducing chemical leaching and fertilizer use. However, more studies are needed to address social adaptability, trust, privacy, and economic income uncertainty, especially in SMEs or in businesses or nations with lower resources; this will require time for adaptation to make the transition away from human ecology. For agriculture to be ESG-sustainable, the deployment of I4FA could be an answer with the support of an open-minded dialogue platform with ESG-minded leaders to complement sustainable agroecosystems on a global scale.

Effect of diisobutyl adipate on the expression of biomarker genes that respond to endocrine disruption and on gonadal sexual differentiation in Japanese medaka (Oryzias latipes).

Phthalate and non-phthalate plasticizers are used in polymer materials, such as plastic and rubber. It has recently been found that diisobutyl adipate (DIBA), which is considered an environmentally safe non-phthalate plasticizer, potentially acts as a thyroid disruptor in fish. Here, we investigated the sexual hormone effects of DIBA based on the expression levels of genes that respond to endocrine disruption and sexual hormone activity in the livers and gonads, and on gonadal sexual differentiation in Japanese medaka. Compared with the control group, the mRNA expression of chgH, vtg1, vtg2, and esr1 was significantly suppressed in the livers of DIBA exposed XX individuals. Furthermore, the mRNA expression of gsdf was significantly upregulated and downregulated in the gonads of XX and XY individuals, respectively. The mRNA expressions of esr1 and esr2b were significantly suppressed by DIBA exposure in the gonads of both XX and XY individuals. These observations suggest that DIBA has potential androgenic activity in Japanese medaka. However, normal testes and ovaries were observed in respective XY and XX medaka after DIBA exposure; therefore, these results suggest that DIBA may have weak androgenic activity.

Anthropogenic Microparticles in Sea-Surface Microlayer in Osaka Bay, Japan.

The abundance, distribution, and composition of microparticles (MPs) in the sea-surface microlayer (S-SML, less than 100 µm of sea surface in this experiment) and in bulk water (1 m under the sea surface) were investigated to evaluate the pollution level of MPs in Osaka Bay in Japan. Both seawater fractions were collected at eight sites including ship navigation routes, the coastal area, and the center of Osaka Bay for 2021-2023. MPs were filtered for four size ranges (10-53, 53-125, 125-500, and >500 µm) and then digested with H2O2. MPs' abundance was microscopically assessed; and polymer types of MPs were identified by a Fourier transform infrared spectrometer (FTIR). For the 22 collections performed along eight sites, the average MPs' abundance was 903 ± 921 items/kg for S-SML, while for the 25 collections performed along the same sites, the average MPs' abundance was 55.9 ± 40.4 items/kg for bulk water, respectively. MPs in both S-SML and bulk water exhibited their highest abundance along the navigation routes. The smallest MPs (10-53 µm) accounted for 81.2% and for 62.2% of all MPs in S-SML and in bulk water among all sites, respectively. Polymethyl methacrylate (PMMA) was the major type of MPs identified while minor ones were polyethylene, polyesters, polystyrene, polypropylene, polyvinyl chloride, polyamide, etc. PMMA comprised 95.1% of total MPs in S-SML and 45.6% of total MPs in bulk water. In addition, PMMA accounted for 96.6% in S-SML and 49.5% in bulk water for the smallest MP category (10-53 µm). It can be assumed that the MP sources were marine paints-primarily APPs (antifouling paint particles)-as well as land coatings. Sea pollution due to microparticles from ship vessels should be given proper attention.

Comment on Peycheva et al. Trace Elements and Omega-3 Fatty Acids of Wild and Farmed Mussels (Mytilus galloprovincialis) Consumed in Bulgaria: Human Health Risks. Int. J. Environ. Res. Public Health 2021, 18, 10023.

First of all, the interesting paper by Peycheva et al. [...].

Utilizing mollusk soft tissue and shells as biomarkers for monitoring heavy metal pollution in mangrove forests.

The primary objective of the study was to examine the distribution of various elements, namely Cadmium (Cd), Copper (Cu), Iron (Fe), Nickel (Ni), and Lead (Pb), in the soft tissues, shells, and associated surface sediments of Cerithidea obtusa (C. obtusa) mangrove snails collected from Sungai Besar Sepang. To conduct the analysis, the preferred and most convenient methods employed were Instrumental Neutron Activation Analysis (INAA) and Atomic absorption spectrometry (AAS). The results showed that the mean concentration of elements in the sediments and soft tissues followed the order Fe > Cu > Ni > Pb > Cd, while for the shell of C. obtusa, it was Fe > Ni > Cu > Pb > Cd.•Iron (Fe) showed the highest concentration among all elements monitored in sediments, soft tissues, and shells of C. obtusa.•The PF results indicated higher incorporation of Pb and Ni into shells.•BSAF results showed that C. obtusa shells accumulated more Cu and Cd from sediments, making them effective biomonitors.

Lower Health Risks of Potentially Toxic Metals after Transplantation of Aquacultural Farmed Mussels from a Polluted Site to Unpolluted Sites: A Biomonitoring Study in the Straits of Johore.

The present field-based study aimed to determine the levels of six potentially toxic metals (PTM)s (Cd, Cu, Fe, Ni, Pb, and Zn determined using a flame atomic-absorption spectrophotometer) using transplanted green-lipped mussel Perna viridis from a polluted site at Kampung Pasir Puteh (KPP) to unpolluted sites at Kampung Sungai Melayu (KSM) and Sungai Belungkor (SB) in the Johore Straits (SOJ), and to estimate the human health risks of the PTMs after the depuration periods. Interestingly, after 10 weeks of depuration in the two unpolluted sites, there were 55.6-88.4% and 51.3-91.7% reductions of the six PTMs after transplantation from KPP to SB and KSM, respectively. Lower risks of health assessments were recorded and judged on the present findings of significantly (p < 0.05) lower levels of safety guidelines, significantly (p < 0.05) lower values of target hazard quotient, and significantly (p < 0.05) lower values of estimated weekly intake, of all the six PTMs after 10 weeks of depuration of the transplanted polluted mussels to the two unpolluted sites in the SOJ. Thus, further reducing the noncarcinogenic risks of the PTMs to the consumers. From an aquacultural point of view, this depuration technique can be recommended to reduce the health risks of PTMs to mussel consumers.

What you need to know about: imaging in patients with renal failure.

Contrast-enhanced medical imaging is commonly requested in clinical practice. Contrast media provide better differentiation of tissue enhancement, improves the soft tissue contrast resolution, and enhances the ability to study the physiology and function of the organs and/or systems. However, contrast media may cause complications, especially in patients with renal failure. This article discusses the use of contrast media in common imaging modalities and the relationship between contrast media and renal function. Administration of iodinated contrast media in computed tomography may cause contrast-associated acute kidney injury; the risk factors and preventive strategies for this are elaborated in this article. Administration of gadolinium-based contrast media in magnetic resonance imaging may lead to nephrogenic systemic fibrosis. Therefore, precautions should be taken when planning for medical imaging for patients with pre-existing acute kidney injury or end-stage chronic kidney disease, for whom contrast media administration in computed tomography or magnetic resonance imaging may be relatively contraindicated. Alternatively, ultrasound contrast agents can be safely used in patients with acute kidney injury or chronic kidney disease. Clinical teams should discuss these patients with radiologists, taking into account the risk-benefits of contrast media, to determine the optimal imaging protocol or modality to answer the clinical query.

Pyriproxyfen influences growth as well as thyroid hormone-related and gh/igf-1 gene expression during the early life stage of zebrafish (Danio rerio).

Pyriproxyfen is an agricultural chemical pesticide that has been detected in the aquatic environment. This study aimed to clarify the effects of pyriproxyfen on the growth as well as thyroid hormone- and growth-related gene expression of zebrafish (Danio rerio) during its early life stage. Pyriproxyfen exhibited a lethal effect in a concentration-dependent manner: the lowest and no effect concentrations were 250.7 and 111.7 µg/L, respectively. These concentrations were considerably higher than the residual environmental concentrations, indicating the low risk of this pesticide when present at such concentrations. In the zebrafish group treated with 56.6 µg/L pyriproxyfen, thyroid hormone receptor ß gene expression levels remained unchanged, whereas thyroid-stimulating hormone ß subunit, iodtyronin deiodinase 2, and thyroid hormone receptor α gene expression levels significantly decreased compared with the control group expression levels. In zebrafish treated with 111.7 or 250.7 µg/L pyriproxyfen, iodtyronin deiodinase 1 gene expression levels significantly increased. These results indicate that pyriproxyfen disrupts thyroid hormone activity in zebrafish. Furthermore, pyriproxyfen exposure inhibited zebrafish growth; consequently, we examined the expression of growth hormone (gh) and insulin-like growth factor-I (igf-1), which are important for growth. Pyriproxyfen exposure suppressed gh expression; however, the igf-1 expression levels remained unchanged. Therefore, growth inhibition due to pyriproxyfen exposure was attributed to the suppression of gh expression.

Byssus of Green-Lipped Mussel Perna viridis as a Biomonitoring Biopolymer for Zinc Pollution in Coastal Waters.

The present study aimed to confirm the use of the byssus (BYS) of the green-lipped mussel Perna viridis as a biomonitoring biopolymer for zinc (Zn) by comparing it to copper (Cu) and cadmium (Cd) pollution in coastal waters under experimental field conditions, based on the transplantation of caged mussels between polluted and unpolluted sites in the Straits of Johore (SOJ). Four important evidential points were found in the present study. First, the 34 field-collected populations with BYS/total soft tissue (TST) ratios > 1 indicated that the BYS was a more sensitive, concentrative, and accumulative biopolymer for the three metals than TST. Significant (p < 0.05) and positive correlations between BYS and TST in terms of the levels of the three metals were observed. Second, the data obtained in the present study were well-supported by the interspecific comparison, which indicated that the BYS of P. viridis was a significantly better biomonitoring biopolymer for the identification of coastal areas exposed to Zn, Cd, and Cu pollution and played the role of an excretion route of metal wastes. Third, the higher positive correlation coefficients for the metals between the BYS sedimentary geochemical fractions than the TST sedimentary geochemical fractions indicated that the BYS was more reflective of metal bioavailability and contamination in coastal waters. Fourth, and most importantly, the field-based cage transplantation study clearly indicated the accumulation and elimination of the three metals by the BYS in both polluted and unpolluted sites in the Straits of Johore. In sum, the BYS of P. viridis was confirmed as a better biopolymer than TST for Zn, as well as Cd and Cu, bioavailability and contamination in tropical coastal waters.

Biomonitoring-Health Risk Nexus of Potentially Toxic Metals on Cerithidea obtusa: A Biomonitoring Study from Peninsular Malaysia.

The present study aimed to assess the human health risks of six potentially toxic metals (PTMs) (Cd, Cu, Fe, Ni, Pb and Zn) in 21 populations of popular mangrove snails, Cerithidea obtusa, collected from Malaysia. In general, the concentrations (mg/kg wet weight) of Cd (0.03-2.32), Cu (11.4-35.2), Fe (40.9-759), Ni (0.40-6.14), Pb (0.90-13.4) and Zn (3.11-129) found in the snails in all populations were lower than the prescribed maximum permissible limits (MPL)s for Cd, Cu, Ni, Pb and Zn. However, in the investigated snail populations, Cd (14%), Pb (62%), Cu (19%), and Zn (10%) were found in exceedance of the MPL of the respective metal. The target hazard quotient (THQ) values in all populations for Cu, Ni, Fe and Zn were all found to be below 1.00. However, for the THQ values of Cd and Pb, two populations exceeded 1.00, while others were below the threshold level. The estimated weekly intake (EWI) of all six metals for all populations was only 0.03-4.65% of the provisional tolerable weekly intake. This conclusively indicates that, based on the EWI, there are no health risks of the six PTMs in the consumption of snails from Malaysia since the assessments are dependent on the consumers' body weight and consumption rate. Nonetheless, the present results indicate that the amounts of snails consumed should be limited to minimize the potential health risks of PTMs to consumers. The relatively low and weak but positive correlations of Cu, Ni, Pb and Zn between C. obtusa and their habitat sediments indicate that C. obtusa can be a potential biomonitor for Cu, Ni, Pb and Zn. This is important for effective mangrove management from the perspective of the sustainable resources from the intertidal mangrove environment. Hence, the biomonitoring-health risk nexus of PTMs in mangrove snails is proposed in the present study.

Effective Microorganisms as Halal-Based Sources for Biofertilizer Production and Some Socio-Economic Insights: A Review.

This paper aims to review the literature on 'Effective Microorganism (EM)' and 'Fertilizer' from the Scopus database and to discuss EMs using Halal-based sources for biofertilizer production from socio economic insights. Based on EM and fertilizer publications on the Scopus database, all the 17 papers reviewed provided no detailed information on the Halal-status of the biofertilizers inoculated with EM. The impacts of Halal-certified biofertilizers will trigger the Halal certification in food products by (a) catering for the increasing Halal food demand due to expectedly Muslim population expansion, (b) contributing to the sustainable buying behaviour of Halal products' consumers in the future, (c) catering for the increasing number of Muslim travellers around the world, (d) becoming a positive driver for higher production of more Halal foods that can enhance food safety, human health and well-being, and (e) creating a cost-effective and increasing food marketability. The later three points (c, d and e) play a very important role in a country's societal well-being and economic growth and development. Although Halal-status is not a must for the world's food marketing, Halal-certified biofertilizer for the Halal-status of food carries the greatest potential to enter the ever-expanding Muslim markets. Finally, it is postulated that the successful usage of EM using Halal-based sources for biofertilizer production will result in two major outcomes from the points of United Nations' Sustainable Development Goals # 9 (Industry, Innovation and Infrastructure) and # 12 (Responsible Consumption and Production). Hence, the presented review provides a starting point for future research considering sustainability and innovation as priorities.

Effects of plasticizer diisobutyl adipate on the Japanese medaka (Oryzias latipes) endocrine system.

Plasticizer pollution of the water environment is one of the world's most serious environmental issues. Phthalate plasticizers can disrupt endocrine function in vertebrates. Therefore, this study analyzed thyroid-related, reproduction-related, and estrogen-responsive genes in Japanese medaka (Oryzias latipes) to determine whether non-phthalate diisobutyl adipate (DIBA) plasticizer could affect endocrine hormone activity or not. Developmental toxicity during fish embryogenesis was also evaluated. At a concentration of 11.57 mg/l, embryonic exposure to DIBA increased the mortality rate. Although abnormal development, including body curvature, edema, and lack of swim bladder inflation, was observed at 3.54 and 11.57 mg/l DIBA, growth inhibition and reduced swimming performance were also observed. In addition, DIBA exposure increased the levels of thyroid-stimulating hormone beta-subunit (tshß) and deiodinase 1 (dio1) but decreased the levels of thyroid hormone receptor alpha (trα) and beta (trß). These results suggest that DIBA has thyroid hormone-disrupting activities in fish. However, kisspeptin (kiss1 and kiss2), gonadotropin-releasing hormone (gnrh1), follicle-stimulating hormone beta (fshß), luteinizing hormone beta (lhß), choriogenin H (chgH), and vitellogenin (vtg1) expression did not change dose-dependently in response to DIBA exposure, whereas gnrh2 and vtg2 expression was elevated. These results indicate that DIBA has low estrogenic activity and does not disrupt the endocrine reproduction system in fish. Overall, this is the first report indicating that non-phthalate DIBA plasticizer is embryotoxic and disrupt thyroid hormone activity in fish.

Effects of non-phthalate plasticizer bis(2-ethylhexyl) sebacate (DEHS) on the endocrine system in Japanese medaka (Oryzias latipes).

Water pollution due to plasticizers is one of the most severe environmental problems worldwide. Phthalate plasticizers can act as endocrine disruptors in vertebrates. In this study, we investigated whether the non-phthalate bis(2-ethylhexyl) sebacate (DEHS) plasticizer can act as an endocrine disruptor by evaluating changes in the expression levels of thyroid hormone-related, reproduction-related, and estrogen-responsive genes of Japanese medaka (Oryzias latipes) exposed to the plasticizer. Following the exposure, the gene expression levels of thyroid-stimulating hormone subunit beta (tshß), deiodinase 1 (dio1), and thyroid hormone receptor alpha (trα) did not change. Meanwhile, DEHS suppressed dio2 expression, did not induce swim bladder inflation, and eventually reduced the swimming performance of Japanese medaka. These findings indicate that DEHS can potentially disrupt the thyroid hormone-related gene expression and metabolism of these fish. However, exposure to DEHS did not induce changes in the gene expression levels of kisspeptin 1 (kiss1), gonadotropin-releasing hormone (gnrh), follicle-stimulating hormone beta (fshß), luteinizing hormone beta (lhß), choriogenin H (chgH), and vitellogenin (vtg) in a dose-dependent manner. This is the first report providing evidence that DEHS can disrupt thyroid hormone-related metabolism in fish.

The Role of Interventional Radiology in the Diagnosis and Treatment of Pulmonary Arteriovenous Malformations.

Pulmonary arteriovenous malformations (PAVMs) are uncommon, predominantly congenital direct fistulous connections between the pulmonary arteries and pulmonary veins, resulting in a right to left shunt. Patients with PAVMs are usually asymptomatic with lesions detected incidentally when radiological imaging is performed for other indications. In this review, we discuss the classification and radiological features of PAVMs as well as their treatment and follow-up options, with a particular focus on percutaneous endovascular techniques and the evolution of the available equipment for treatment.

Correction: Yap, C.K.; Al-Mutairi, K.A. Copper and Zinc Levels in Commercial Marine Fish from Setiu, East Coast of Peninsular Malaysia. Toxics 2022, 10, 52.

The published article [...].

Thyroid hormone disruption by bis-(2-ethylhexyl) phthalate (DEHP) and bis-(2-ethylhexyl) adipate (DEHA) in Japanese medaka Oryzias latipes.

Pollution of water bodies with plasticizers is a serious environmental problem worldwide. In this study, we investigated the effects of plasticizers bis-(2-ethylhexyl) phthalate (DEHP) and bis-(2-ethylhexyl) adipate (DEHA) in Japanese medaka (Oryzias latipes). DEHP significantly increased the expression of all the genes tested: thyroid stimulating hormone beta subunit (tshß-like), tshß, deiodinase 1 (dio1), deiodinase 2 (dio2), and thyroid hormone receptor alpha (trα) and beta (trß). However, DEHA only significantly increased tshß at 7.4 µg/L but significantly decreased dio2 expression at 25.8, 111.1, and 412.6 4 µg/L, while other genes were not significantly affected. Both chemicals reduced eye size and total body length, but did not affect embryo development, hatching time and rate, and swimming performance. DEHA alone affected swim bladder inflation and not DEHP. This is the first report that not only DEHP but also DEHA disrupt thyroid hormone activity in fish. DEHP contamination (13.2 µg/L) was detected in tap water from Kobe, Japan; thus, tap water itself may disrupt thyroid hormone activity in Japanese medaka. Importantly, the effective concentration of DEHP for thyroid hormone-related gene expression and growth was close to or lower than DEHP concentrations reported in surface water elsewhere, indicating that DEHP contamination is a serious aquatic pollution.

Source apportionment and health risk assessment of polycyclic aromatic hydrocarbons (PAHs) in the coastal ecosystem of the Brunei Bay, Brunei.

Pollution characteristics and associated ecological risks of PAHs in sediments from Brunei Bay, Brunei were investigated. The concentrations of ∑16 PAHs ranged from 826.7 to 2955.3 µg kg-1, indicating moderate to high level of pollution. Source apportionment of PAHs by molecular isomeric ratios and positive matrix factorization model indicated impact of potential anthropogenic PAH sources including combustion of biomass and fossil fuels. The data indicated relatively no significant ecotoxicological risk for most of PAH compounds. To estimate the individual c-PAH toxicity, the toxic equivalent quantity (c-TEQ) was calculated. Results of the TEQ analysis showed that BaP followed by DBA and BaA are the most carcinogenic of PAHs examined in the study area. The evaluation of human health risk of PAHs revealed that the cancer risk of PAHs for adults and children was higher than the USEPA threshold (<1E-06) and lower than 1E-3, implying low to moderate risk.

Galectin-1 orchestrates an inflammatory tumor-stroma crosstalk in hepatoma by enhancing TNFR1 protein stability and signaling in carcinoma-associated fibroblasts.

Most cases of hepatocellular carcinoma (HCC) arise with the fibrotic microenvironment where hepatic stellate cells (HSCs) and carcinoma-associated fibroblasts (CAFs) are critical components in HCC progression. Therefore, CAF normalization could be a feasible therapy for HCC. Galectin-1 (Gal-1), a ß-galactoside-binding lectin, is critical for HSC activation and liver fibrosis. However, few studies has evaluated the pathological role of Gal-1 in HCC stroma and its role in hepatic CAF is unclear. Here we showed that Gal-1 mainly expressed in HCC stroma, but not cancer cells. High expression of Gal-1 is correlated with CAF markers and poor prognoses of HCC patients. In co-culture systems, targeting Gal-1 in CAFs or HSCs, using small hairpin (sh)RNAs or an therapeutic inhibitor (LLS30), downregulated plasminogen activator inhibitor-2 (PAI-2) production which suppressed cancer stem-like cell properties and invasion ability of HCC in a paracrine manner. The Gal-1-targeting effect was mediated by increased a disintegrin and metalloprotease 17 (ADAM17)-dependent TNF-receptor 1 (TNFR1) shedding/cleavage which inhibited the TNF-α → JNK → c-Jun/ATF2 signaling axis of pro-inflammatory gene transcription. Silencing Gal-1 in CAFs inhibited CAF-augmented HCC progression and reprogrammed the CAF-mediated inflammatory responses in a co-injection xenograft model. Taken together, the findings uncover a crucial role of Gal-1 in CAFs that orchestrates an inflammatory CSC niche supporting HCC progression and demonstrate that targeting Gal-1 could be a potential therapy for fibrosis-related HCC.

The impact assessment of desalination plant discharges on heavy metal pollution in the coastal sediments of the Persian Gulf.

In recent decades the development of desalination plants (DPs) for desalination of seawater has increased dramatically, while little attention has been paid to the effects of this activity on the accumulation of heavy metals (HMs) in the sediments of affected ecosystems. The present study was implemented to evaluate (1) heavy metal accumulation in sediments impacted by DPs discharges, (2) spatial and temporal changes of HMs and the contamination degree by different types of pollution indexes (single and integrated indices), and (3) ecological risk assessment of cadmium (Cd), lead (Pb), zinc (Zn) and copper (Cu) in sediments affected by DPs discharges. A total of 288 sediment samples were collected seasonally at 24 stations from November 2019 to October 2020. Analysis of HMs concentrations in sediments near the desalination plant discharge provided evidence of local contamination. Maximum concentration of Cu and Pb elements were found in sediments near the desalination plant discharge point. Hierarchical cluster analysis revealed clear segregation of stations impacted by desalination plant discharges and away from discharges. The values of PLI index in sediments of all sampling stations were < 1, indicating that there was no metal pollution by this index. The potential ecological risk index (PERI) ranged from 5.33 ± 0.51 to 11.81 ± 4.98 in sampling sediments and were classified as "low potential ecological risk". These results demonstrate that the DPs discharge increased HMs concentrations in the sediments in close proximity to outlets. The necessary and practical regulations and policies regarding the rejection of the DPs discharge and disposal of chemical compounds must be implemented and enforced.

Immunomodulatory Effect and an Intervention of TNF Signalling Leading to Apoptotic and Cell Cycle Arrest on ORL-204 Oral Cancer Cells by Tiger Milk Mushroom, Lignosus rhinocerus.

Research background: Tiger milk mushroom (Lignosus rhinocerus) is a medicinal mushroom that is geographically distributed in the region of South China, Thailand, Malaysia, Indonesia, Philippines and Papua New Guinea. Consumption of its sclerotium has been reported to treat various ailments. However, its anticancer potential towards oral cancer cell lines is yet to be determined considering the traditional method of its consumption by biting/chewing of the sclerotium. Experimental approach: Mushroom sclerotial powder of cultivar TM02® was extracted and fractionated in a chromatographic column prior to cytotoxicity testing against a panel of human oral cancer cell lines. The capability of the identified bioactive fraction in regulating several molecules associated with its tumour necrosis factor (TNF) pathway was investigated. Results and conclusions: 2,5-Diphenyl-2H-tetrazolium bromide (MTT) proliferation assay indicated that cell lines ORL-48 (derived from gingiva), ORL-188 (derived from the tongue) and ORL-204 (derived from buccal mucosa) were inhibited by cold water extract of L. rhinocerus sclerotia and its high-molecular-mass fraction (HMM) in varying degrees with ORL-204 being most affected. Hence, the treatment of ORL-204 with HMM mushroom extract was further investigated. HMM mushroom extract induced apoptosis and G0/G1 phase cell cycle arrest through caspase-3/7 cleavage. Activities of MIP2 and COX-2 were downregulated by 0.2- and 4.6-fold respectively in the HMM mushroom extract-treated ORL-204 cells. Novelty and scientific contribution: Using ORL-204, we showed that HMM mushroom extract may act via the TNF pathway at various network sites as a potential dietary compound for cancer prevention and natural adjunct therapeutic to conventional cancer treatment.