Red meat consumption is associated with an increased overall cancer risk: a prospective cohort study in Korea.

Cancer is a leading cause of death, and the dietary pattern in Korea is changing rapidly from a traditional Korean diet to a Westernised diet. In the present study, we investigated the effects of dietary factors on cancer risk with a prospective cohort study. Among 26,815 individuals who participated in cancer screening examinations from September 2004 to December 2008, 8024 subjects who completed a self-administered questionnaire concerning demographic and lifestyle factors, and a 3 d food record were selected. As of September 2013, 387 cancer cases were identified from the National Cancer Registry System, and the remaining individuals were included in the control group. The hazard ratio (HR) of cancer for the subjects older than or equal to 50 years of age was higher (HR 1.80, 95% CI 1.41, 2.31; P< 0.0001) than that for the other subjects. Red meat consumption, Na intake and obesity (BMI ≥ 25 kg/m²) were positively associated with overall cancer incidence in men (HR 1.41, 95% CI 1.02, 1.94; P= 0.0382), gastric cancer (HR 2.34, 95% CI 1.06, 5.19; P= 0.0365) and thyroid cancer (HR 1.56, 95% CI 1.05, 2.31; P= 0.0270), respectively. Participants who had at least three dietary risk factors among the high intakes of red meat and Na, low intakes of vegetables and fruits, and obesity suggested by the World Cancer Research Fund/American Institute for Cancer Research at baseline tended to have a higher risk of cancer than the others (HR 1.26, 95% CI 0.99, 1.60; P= 0.0653). In summary, high intakes of red meat and Na were significant risk factors of cancer among Koreans.

Editing of the ethylene biosynthesis gene in carnation using CRISPR-Cas9 ribonucleoprotein complex.

The study aimed to edit ethylene (ET) biosynthesis genes [1-aminocyclopropane-1-carboxylic acid (ACC) synthetase 1 (ACS1) and ACC oxidase 1 (ACO1)] in carnation using the CRISPR/Cas9 ribonucleoprotein (RNP) complex system. Initially, the conserved regions of the target genes (ACS1 and ACO1) were validated for the generation of different single guide RNAs (sgRNAs), followed by the use of an in vitro cleavage assay to confirm the ability of the sgRNAs to cleave the target genes specifically. The in vitro cleavage assay revealed that the sgRNAs were highly effective in cleaving their respective target regions. The complex of sgRNA: Cas9 was directly delivered into the carnation protoplast, and the target genes in the protoplast were deep-sequenced. The results revealed that the sgRNAs were applicable for editing the ET biosynthesis genes, as the mutation frequency ranged from 8.8 to 10.8% for ACO1 and 0.2-58.5% for ACS1. When sequencing the target genes in the callus derived from the protoplasts transformed with sgRNA: Cas9, different indel patterns (+ 1, - 1, and - 8 bp) in ACO1 and (- 1, + 1, and + 11) in ACS1 were identified. This study highlighted the potential application of CRISPR/Cas9 RNP complex system in facilitating precise gene editing for ET biosynthesis in carnation.

Protoplast isolation and transient gene expression in different petunia cultivars.

The protocol optimized for Petunia hybrida cv. Mirage Rose produced high protoplast yields in 3 out of other 11 cultivars (Damask White, Dreams White, and Opera Supreme White). Factors optimized in the protoplast transfection process showed that the best transfection efficiency (80%) was obtained using 2.5 × 105 protoplast density, 40% polyethylene glycol (PEG) concentration, 10 µg plasmid DNA, and 15 min of transfection time. Assessing the usability of the protocol for other cultivars (Damask White, Dreams White, and Opera Supreme White), a reasonable protoplast transfection efficiency (⁓50%) was observed in the cultivars Dreams White and Opera Supreme White, with lower efficiency (⁓50%) observed in the cv. Damask White. The transient expression of enhanced green fluorescent protein (eGFP) in the nucleus of the transfected protoplasts of all cultivars was confirmed using PCR. This system could be valuable for genome editing of unwanted genes in petunias using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) technology. Furthermore, it could contribute to other studies on protein subcellular localization, protein-protein interactions, and functional gene expression in the petunias.

Loss of ACO4 in petunia improves abiotic stress tolerance by reducing the deleterious effects of stress-induced ethylene.

To investigate the role of ethylene (ET) in abiotic stress tolerance in petunia cv. 'Mirage Rose', petunia plants in which the ET biosynthesis gene 1-aminocyclopropane-1-carboxylic acid oxidase 4 (ACO4) was knocked out (phaco4 mutants) and wild-type (WT) plants were exposed to heat and drought conditions. Loss of function of ACO4 significantly delayed leaf senescence and chlorosis under heat and drought stress by maintaining the SPAD values and the relative water content, indicating a greater stress tolerance of phaco4 mutants than that of WT plants. This tolerance was related to the lower ET and reactive oxygen species levels in the mutants than in WT plants. Furthermore, the stress-induced expression of genes related to ET signal transduction, antioxidant and proline activities, heat response, and biosynthesis of abscisic acid was higher in the mutants than in WT plants, indicating a greater stress tolerance in the former than in the latter. These results demonstrate the deleterious effects of stress-induced ET on plant growth and provide a better physiological and molecular understanding of the role of stress ET in the abiotic stress response of petunia. Because the loss of function of ACO4 in petunia improved stress tolerance, we suggest that ACO4 plays a vital role in stress-induced leaf senescence and acts as a negative regulator of abiotic stress tolerance.

Role of Ethylene Biosynthesis Genes in the Regulation of Salt Stress and Drought Stress Tolerance in Petunia.

Ethylene plays a critical signaling role in the abiotic stress tolerance mechanism. However, the role of ethylene in regulating abiotic stress tolerance in petunia has not been well-investigated, and the underlying molecular mechanism by which ethylene regulates abiotic stress tolerance is still unknown. Therefore, we examined the involvement of ethylene in salt and drought stress tolerance of petunia using the petunia wild type cv. "Merage Rose" and the ethylene biosynthesis genes (PhACO1 and PhACO3)-edited mutants (phaco1 and phaco3). Here, we discovered that editing PhACO1 and PhACO3 reduced ethylene production in the mutants, and mutants were more sensitive to salt and drought stress than the wild type (WT). This was proven by the better outcomes of plant growth and physiological parameters and ion homeostasis in WT over the mutants. Molecular analysis revealed that the expression levels of the genes associated with antioxidant, proline synthesis, ABA synthesis and signaling, and ethylene signaling differed significantly between the WT and mutants, indicating the role of ethylene in the transcriptional regulation of the genes associated with abiotic stress tolerance. This study highlights the involvement of ethylene in abiotic stress adaptation and provides a physiological and molecular understanding of the role of ethylene in abiotic stress response in petunia. Furthermore, the finding alerts researchers to consider the negative effects of ethylene reduction on abiotic stress tolerance when editing the ethylene biosynthesis genes to improve the postharvest quality of horticultural crops.

Occurrence and risk characterization of non-nutritive sweeteners in selected food products from Korea.

Non-nutritive sweeteners (NNS) are food additives that impart a sweet taste to food product with lower calories. Acesulfame potassium, aspartame, sodium saccharin, sucralose, steviol glycosides and enzymatically modified stevia are permitted in Korea. The study established the method of each NNS and applied it to each food items consumed in Korea. For risk assessments, the estimated daily intake (EDI) value for each NNS was calculated. EDI values of NNS were compared directly with each ADI (acceptable daily intake). The total estimated daily intake ranges by age compared with the % ADI were 0.12-0.53, 0.93-1.68, 0.05-0.20, 0.06-0.42 and 0.17-0.98% for acesulfame potassium, sodium saccharin, aspartame, sucralose and sum of stevioside and rebaudioside A, which were based on the overall averages. It can be concluded that the daily dietary intake of each of the five NNS is at a safe level when considered as a proportion of the ADI. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-01012-9.

Paeonia lactiflora Root Extract and Its Components Reduce Biomarkers of Early Atherosclerosis via Anti-Inflammatory and Antioxidant Effects In Vitro and In Vivo.

Although various physiological activities of compounds obtained from Paeonia lactiflora have been reported, the effects of P. lactiflora extract (PLE) on early atherosclerosis remain unclear. Therefore, in this study, we investigated the in vitro and in vivo antiatherosclerosis and in vitro antioxidant effects of PLE and its compounds. PLE suppresses the tumor necrosis factor (TNF)-α-induced capacity of THP-1 cells to adhere to human umbilical vein endothelial cells (HUVECs), vascular cell adhesion molecule (VCAM)-1 expression, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling in HUVECs. PLE also suppresses TNF-α-induced nuclear translocation of NF-κB p65 from cytosol as well as the enhanced TNFA and C-C motif chemokine ligand 2 (CCL2) mRNA expression in HUVECs. We identified and quantified the following PLE compounds using high-performance liquid chromatography with diode array detection: methyl gallate, oxypaeoniflorin, catechin, albiflorin, paeoniflorin, benzoic acid, benzoylpaeoniflorin, and paeonol. Among these, methyl gallate had the strongest inhibitory effect on monocyte adherence to TNF-α-induced HUVECs and the VCAM-1 expression. Reverse transcriptase real-time quantitative polymerase chain reaction showed that PLE compounds had a dissimilar inhibition effect on TNF-α-induced mRNA expression levels of CCL2, TNFA, and IL6 in HUVECs. Except for paeonol, the compounds inhibited lipopolysaccharide (LPS)-induced reactive oxygen species production in RAW264.7 cells. In vivo, oral administration of PLE improved TNF-α-induced macrophage infiltration to the vascular endothelium and expression of VCAM-1, as well as IL6 and TNFA gene expression in the main artery of mice. PLE could be useful as a nutraceutical material against early atherosclerosis via the combined effects of its components.

Validation and measurement uncertainty of GC-MS quantification method for nicotine in dried mushrooms using QuEChERS extraction.

Nicotine is an alkaloid and a secondary plant metabolite that has been used as an insecticide. Despite their widespread application, the EU banned the use of nicotine-containing pesticides in December 2008. However, studies in Europe have found nicotine in mushrooms. Nicotine has also been detected in wild mushrooms, so there are other causes of contamination as well as pesticide. This study reports the development of GC-MS method for quantitatively analysing nicotine in mushrooms. This method provides recoveries of 89.5-92.5%, intra-day precisions of 0.32-0.85%, and inter-day precisions of 0.73-2.36%, with limits of detection and quantification of 0.38 and 1.15 µg kg-1, respectively. The relative expanded uncertainty result of 2.8-4.0% complies with CODEX requirements. The method was successfully applied to eleven mushroom samples in which nicotine was detected at levels of 0.033-1.713 mg kg-1. Therefore, this method is suitable for the quantification of nicotine in dried mushrooms to ensure pre-emptive food safety.

Application of Non-Destructive Rapid Determination of Piperine in Piper nigrum L. (Black Pepper) Using NIR and Multivariate Statistical Analysis: A Promising Quality Control Tool.

Piperine is a bioactive alkaloid compound which provides a unique spicy flavor derived from plants of the Piper nigrum L. Black pepper (n = 160) collected from Vietnam was studied using non-destructive near infrared spectroscopy (NIRS). The spectral acquisition ranged from 1100 to 2500 nm, and a chemometrics analysis program was performed to quantify the piperine contents. High performance liquid chromatography (HPLC) analysis was carried out to develop a chemometric model based on reference values. The black pepper samples were divided into two groups used for calibration (n = 120) and prediction (n = 40) sets. The optimum calibration model was developed by pretreatment of the spectra. The analyses results based on the prediction samples included a coefficient of determination (R2) of 0.914, a root mean square error of prediction (RMSEP) and a standard error of prediction (SEP) of about 0.220 g/100 g, and a ratio performance to deviation (RPD) value of 3.378 regarding the partial least square (PLS) regression model, and an R2 of 0.921, an RMSEP and SEP of 0.210 g/100 g, and an RPD of 3.571, with respect to the principal components (PC) regression model. These results indicate that NIRS can be applicable as a control, or as an alternative rapid and effective method to quantify piperine in P. nigrum L.

Feasibility of rapid piperine quantification in whole and black pepper using near infrared spectroscopy and chemometrics.

Piperine is a bioactive alkaloid that possesses various health benefits and is responsible for the pungent aroma of pepper. Piperine content in whole and ground black pepper (n = 132) was analyzed by near-infrared spectroscopy (NIRS) in the 950 to 1650 nm wavelength window. Chemometric modeling using partial least square regression was performed, and outliers were checked and removed during the preparation of the calibration curve by considering sample residual variance and sample leverage. Model accuracy was evaluated with a low root-mean-square error of cross-validation (RMSECV) and a high ratio performance to deviation (RPD). The optimal model had a coefficient of determination (R2 ) of 0.726, RMSECV of 0.289 g/100 g, and RPD of 1.744 for the whole black pepper. The results of R2 , RMSECV, and RPD for the ground black pepper were 0.850, 0.231 g/100 g, and 2.424, respectively. Therefore, based on the perspective of onsite process, the proposed NIRS method can be employed for selecting abnormal samples during the inspection of black pepper raw material and for quantifying piperine contents of final black pepper product. PRACTICAL APPLICATION: Generally, the quality indicators of black pepper are graded solely based on their external appearance, quality, and size. This study discloses the development of a near-infrared spectroscopy-based fast and accurate nondestructive analytical method for the detection of piperine, a bioactive constituent of pepper, as a tool for the quality control of whole and ground black pepper.

The Role of Red Meat and Flavonoid Consumption on Cancer Prevention: The Korean Cancer Screening Examination Cohort.

Markedly increased red meat consumption is a cancer risk factor, while dietary flavonoids may help prevent the disease. The purpose of this study was to investigate the associations of red meat and flavonoid consumption with cancer risk, based on data from 8024 subjects, drawn from the 2004-2008 Cancer Screening Examination Cohort of the Korean National Cancer Center. Hazard ratios (HRs) were obtained by using a Cox proportional hazard model. During the mean follow-up period of 10.1 years, 443 cases were newly diagnosed with cancer. After adjusting for age, there was a significant correlation between cancer risk and the daily intake of ≥43 g of red meat per day (HR 1.31; 95% CI 1.01, 1.71; p = 0.045), and total flavonoid intake tended to decrease cancer risk (HR 0.70; 95% CI 0.49, 0.99; highest vs. lowest quartile; p-trend = 0.073) in men. Following multivariable adjustment, there were no statistically significant associations between flavonoid intake and overall cancer risk in individuals with high levels of red meat intake. Men with low daily red meat intake exhibited an inverse association between flavonoid consumption and cancer incidence (HR 0.41; 95% CI 0.21, 0.80; highest vs. lowest; p-trend = 0.017). Additional research is necessary to clarify the effects of flavonoid consumption on specific cancer incidence, relative to daily red meat intake.

Erratum: The Role of Red Meat and Flavonoid Consumption on Cancer Prevention: The Korean Cancer Screening Examination Cohort; Nutrients 2017, 9, 938.

The authors have requested the following corrections to their paper [...].