Occurrence and health risk assessment of tropane alkaloids in cereal foods consumed in Korea.

Tropane alkaloids (TA) are natural toxins found in certain plants, including cereals, of which atropine and scopolamine are the main species of concern due to their acute toxicity. This study aimed to determine the occurrence of TA in cereal foods and assess the potential health risks associated with their consumption in Korea. TA levels were analyzed in 80 raw and 71 processed cereal samples, which were distributed throughout Korea in 2021, using ultra-performance liquid chromatography-tandem mass spectrometry. At least one of the six TA species, namely atropine, scopolamine, pseudotropine, tropinone, scopine, and 6-hydroxytropinone, was detected in 10 out of the 151 samples at levels ranging from 0.12 to 88.10 µg kg-1. Dietary exposure (mean, 0.23 ng kg-1 bw day-1) to atropine and scopolamine in the Korean population was estimated to be low across all age groups. This is despite considering worst-case scenarios using the total concentrations of atropine and scopolamine in a millet sample, both of which were detected, and 95th percentile consumption for consumers of millet only. Both the hazard index and margin of exposure methods indicated that the current levels of TA exposure from millet consumption were unlikely to pose significant health risks to the Korean population.

Detection method for genetically modified potato using an ultra-fast PCR system.

Genetically modified (GM) potatoes having resistance to insects and viral diseases, low reducing sugar contents, and black spots for high quality continue to be developed. However, no GM potato has been approved as food or feed in the Republic of Korea as the country adheres to a zero-tolerance policy to unauthorized genetically modified organisms (GMOs). When the self-sufficiency rate is low, a detection method to assess GMOs in crops or other products is necessary. Therefore, a rapid method for two GM potato events (SPS-Y9 and EH92-527-1) using an ultra-fast PCR (UF-PCR) system has been developed, and its specificity, sensitivity, and applicability were demonstrated. UF-PCR can decrease the runtime of PCR by more than half of that needed in conventional methods. However, UF-PCR is not a common method for GMO analysis. This rapid detection method may be useful for GMO analyses in field conditions.

Risk assessment of hexachlorobutadiene (HCBD) in the Korean diet.

Hexachlorobutadiene (HCBD) is a persistent organic pollutant (POP), and a toxin whose primary target organ is the kidney. Consequently, quantifying the amount of HCBD in food is essential for determining whether it poses a health risk. The current study established and validated an analytical method for assessing HCBD in food using gas chromatography/mass spectrometry (GC/MS). Subsequently, we conducted for the first time a risk assessment of HCBD through Korean food consumption. The ranges of HCBD concentration in 595 food samples were as follows: not detectable (nd)-0.947 ng/g for agricultural products, nd-0.920 ng/g for animal products, nd-1.323 ng/g for fishery products and nd-1.081 ng/g for processed food products. The daily intakes of HCBD for the general population were 0.22 ng/kg body weight (b.w.)/day for agricultural products, 0.30 ng/kg b.w./day for animal products, 0.07 ng/kg b.w./day for fishery products and 0.33 ng/kg b.w./day for processed food products. These exposure levels are below the tolerable daily intake (TDI, 2 µg/kg body weight/day) established by the National Institute of Food and Drug Safety Evaluation (NiFDS) in the Republic of Korea. Taking into account the risk index of 0.011%, we can, therefore, conclude that there are no health concerns for the Korean population.

Development of a rapid detection method for genetically modified rice using the ultra-fast PCR system.

Genetically modified (GM) rice varieties containing traits such as tolerance to abiotic stress and resistance against pests and diseases continue to be developed. However, contamination incidents from unauthorized GM rice varieties have been encountered. To date, no GM rice crop has been authorized for consumption and/or commercialization in Korea. Therefore, to enhance safety management of unauthorized genetically modified organisms (GMOs), accurate and reliable detection methods are needed to identify GMOs in crops or products. In this study, we developed rapid detection methods for GM rice events (Bt63, KMD1, Kefeng6, Kefeng8, and LLRice62) using ultra-fast PCR system. Ultra-fast PCR is a state-of-the-art technology and decreases PCR run-times dramatically. However, the ultra-fast PCR is not widely used in GMO analysis. Thus, we designed a detection method for five events of GM rice and confirmed them by performing specificity, sensitivity, and applicability assays. All results demonstrate that the ultra-fast PCR system is a specific, sensitive, and reliable method to identify and monitor GM rice events. Additionally, it can be utilized as a rapid and simple method for GMO analysis in crops or processed products. This study can be used as a reference for future research on new analysis methods of unauthorized GMOs.

Occurrence and risk assessment of sterigmatocystin in agricultural products and processed foods in Korea.

Sterigmatocystin (STC), a carcinogenic mycotoxin, is known to be produced during the biosynthetic pathway of aflatoxin B1. STC in various foods was determined by LC-MS/MS and its risks were assessed. The analytical method was validated in different food categories, and the performance was acceptable based on the criteria of AOAC. A total 1,135 samples (613 agricultural products and 522 processed foods) were analysed, and STC was detected in 46 samples, indicating a detection rate of 4.1%. STC was found in the range of 0.08-10.07 ng/g, and the detection rates of STC were 3.9% in agricultural products and 4.2% in processed foods. The exposure to STC by average consumption of foods was estimated to 0.09 ng/kg b.w./day. The margin of exposure (MOE) approach was applied to assess the risk of STC, and MOE for the whole population was over 1 × 106. Exposure to STC from the consumption of foods distributed in Korea is unlikely to cause human health problems.

Microwave-assisted digestion method using diluted nitric acid and hydrogen peroxide for the determination of major and minor elements in milk samples by ICP-OES and ICP-MS.

A novel method for microwave-assisted digestion of milk samples using diluted HNO3and H2O2 with a single reaction chamber was developed for elemental analysis by ICP-based techniques. The optimal conditions for digestion were 0.25 g of sample mass, 6 mL of 0.1 molL-1HNO3and 2 mL of 30% H2O2 at 250 ℃ and 160 bar. The optimized procedure resulted in low residual carbon content and residual acidity of 260 mgL-1 and 0.06 mol L-1, respectively. The limits of detection ranged from 0.286Õ¸g g-1(Ca) to 82.990Õ¸g g-1(Fe). In addition, the proposed method was considered an excellent green analysis method with a final score of 87 based on the analytical Eco-Scale. Finally, the method was validated and applied to the determination of Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, Se, and Zn in milk samples from South Korea.

Assessment of human estrogen receptor agonistic/antagonistic effects of veterinary drugs used for livestock and farmed fish by OECD in vitro stably transfected transcriptional activation assays.

The presence of veterinary drug residues in foods and the environment could potentially cause adverse effects on humans and wildlife. Several veterinary drugs were reported to exhibit endocrine disrupting effects via binding affinities to sexual hormone receptors such as estrogen and androgen receptors. Therefore, we confirmed the human estrogen receptor (ER) agonistic/antagonistic effects of 135 chemicals that were used as veterinary drugs in Korea by the official Organization for Economic Cooperation and Development (OECD) in vitro ER transcriptional activation (TA) assay using the VM7Luc4E2 cell line. In the case of ER agonist screening, 7 veterinary drugs (cefuroxime, cymiazole, trenbolone, zeranol, phoxim, altrenogest and nandrolone) were determined to be ER agonists. In addition, only zeranol was found to exhibit weak ER antagonistic activity. These 7 veterinary drugs, which were determined as ER agonists and/or antagonists by an OECD in vitro assay, were also found to have binding affinity to ERs. These results indicate that various veterinary drugs possess potential (anti-)estrogenic effects. However, further study is needed to determine the precise endocrine-disrupting effects of these compounds.

Absorption kinetics of vitamin E nanoemulsion and green tea microstructures by intestinal in situ single perfusion technique in rats.

The absorption kinetics of food ingredients such as nanoemulsified vitamin E and green tea microstructures were evaluated by the intestinal in situ single perfusion technique. Absorption rate, sub-acute oral toxicity and organ morphology in a rat model were examined. The intestinal in situ single perfusion technique and HPLC analysis were applied to investigate the absorption rate of selected materials by examining time-dependent changes in the serum levels of catechin and dl-α-tocopherol. The acute toxicity test and histopathological evaluation were applied to analyze the safety of microsized green tea and nanosized vitamin E in a rat model. Total serum dl-α-tocopherol levels significantly increased with nanosized vitamin E administration (P<0.05). Rats treated to nanosized vitamin E until 90min after administration showed significantly increased absorption rate of serum dl-α-tocopherol levels at each time point (10min interval) (P<0.001). Rats administered 2000mg/kg of nanosized vitamin E and microsized green tea did not show signs of acute toxicity or death after 14days of observation. In addition, macroscopic analysis showed that there were no changes in representative organ sections of rats following the oral administration of food-related nanoscale materials. We successfully demonstrated that using nanosized vitamin E increased absorption rate to a greater extent than normal food-related material, and these results occurs via safety analyses on food-related nanoscale materials for human consumption. These results could be useful for the design and development of novel nanoemulsified vitamin E and microsized green tea formulations that can overcome the problem of their bioavailability and improve their efficacy while still maintaining their essential therapeutic efficacies.

Veterinary drug, 17ß-trenbolone promotes the proliferation of human prostate cancer cell line through the Akt/AR signaling pathway.

Trenbolone acetate (TBA) is a synthetic anabolic steroidal growth factor that is used for rapid muscle development in cattle. The absorbed TBA is hydrolyzed to the active form, 17ß-trenbolone (17 TB; 17ß-hydroxy-estra-4,9,11-trien-3-one) in meat and milk products, which can cause adverse health effects in humans. Similar to 5α-dihydrotestosterone (DHT), 17 TB was reported to exhibit endocrine disrupting effects on animals and humans due to its androgenic effect via binding to the androgen receptor. The purpose of this study is to investigate the molecular mechanism of cell proliferation in prostate cancer (PCa) cells treated with 17 TB. We found that 17 TB induces AR-dependent cell proliferation in the human prostate cancer cell line, 22Rv1 in a concentration dependent manner. Treatment with 17 TB increased the expression of cell cycle regulatory proteins, cyclin D2/CDK-4 and cyclin E/CDK-2, whereas the expression of p27 was down-regulated. Furthermore, phosphorylation of Rb and activation of E2F were also induced, which suggests the activation of cyclin D2/CDK-4 and cyclin E/CDK-2 in the cells. When 22Rv1 cells were exposed to 30 pM of 17 TB, which is the effective concentration (EC50) value required to observe proliferative effects on 22Rv1 cells, the expression levels of the phosphorylated forms of Akt and GSK3ß were increased. This study demonstrates that 17 TB induces AR-dependent proliferation through the modulation of cell cycle-related proteins in the Akt signaling pathway. The present study provides an effective methodology for identifying cell proliferation signaling of veterinary drugs that exert AR agonistic effects.

High resolution metabolomics to determines the risk associated with bisphenol A exposure in humans.

Although high BPA exposure has been correlated with several metabolic diseases, the underlying mechanisms are unclear. In the present study, a metabolomics approach was used to explore the metabolic variations caused by low or high BPA exposure in female (n = 96) and male (n = 98) urine. Fatty acid elongation and sphingolipid metabolism were affected by high BPA exposure in males and females. Fatty acid elongation and sphingolipid metabolism were further investigated among age groups consisted of 30-39 yrs old, 40-49 yrs old, and 50-59 yrs old males and females with high or low urinary BPA. High BPA-exposed males in 30 s and females in 40 s were found with significant disturbance in fatty acid elongation and sphingolipid metabolism, respectively. Additionally, females in 40 s showed elevated inflammatory metabolites: 6-ketoprostaglandin E1 and thromboxane. In the present study, we have demonstrated that environmental metabolomics is useful to elucidate the health effects of BPA exposure.

Assessment of androgen receptor agonistic/antagonistic effects on 25 chemicals in household applicants by OECD in vitro stably transfected transcriptional activation assays.

The aim of this study is to assess the androgen receptor (AR) agonistic/antagonistic effects on various chemicals, which are used in household products including cleaning agents and wetted tissues by in vitro OECD test guideline No. 458 (using AR-EcoScreen™ cell line) and the me-too test method (using 22Rv1cell line), which was adopted as OECD project No. 4.99. All chemicals were not determined as AR agonists. However α-dodecyl-ω-hydroxypoly (oxyethylene) and 3-iodo-2-propynyl butylcarbamate have shown a weak AR antagonistic effects with IC50 values of 2.18 ±â€¯0.12 and 4.26 ±â€¯0.17 µg/ml via binding affinity to AR in only 22Rv1/mouse mammary tumor virus using AR transcriptional activation assay, because of their different cytotoxicity on each applied cell line. This report firstly provides information about agonistic/antagonistic effects against human AR of various chemicals including surfactants and biocides by OECD in vitro stably transfected transcriptional activation assays. However, further in vivo and human model studies are needed to confirm their adverse effects.

A DNA replication-related element downstream from the initiation site of Drosophila selenophosphate synthetase 2 gene is essential for its transcription.

Selenophosphate synthetase catalyzes the synthesis of selenophosphate which is a selenium donor for Sec biosynthesis. In Drosophila melanogaster, there are two types of selenophosphate synthetases designated dSPS1 and dSPS2, where dSPS2 is a selenoprotein. The mechanism of gene expression of dSPS2 as well as other selenoproteins in Drosophila has not been elucidated. Herein, we report an essential regulator system that regulates the transcription of the dSPS2 gene (dsps2). Through deletion/substitution mutagenesis, the downstream DNA replication-related element (DRE) located at +71 has been identified as an essential element for dsps2 promoter activity. Furthermore, double-stranded RNA interference (dsRNAi) experiments were performed to ablate transcription factors such as TBP, TRF1, TRF2 and DREF in Schneider cells. The dsRNAi experiments showed that dsps2 promoter activities in DREF- and TRF2-depleted cells were significantly decreased by 90% and 50%, respectively. However, the depletion of TBP or TRF1 did not affect the expression level of dsps2 even though there is a putative TATA box at -20. These results strongly suggest that the DRE/DREF system controls the basal level of transcription of dsps2 by interacting with TRF2.