Contamination of ultraviolet absorbers in food: toxicity, analytical methods, occurrence and risk assessments.

Ultraviolet (UV) absorbers are chemical substances that are widely used as defenses against the damaging effects of solar radiations. UV absorbers, despite their benefits, are categorized as emerging pollutants because they have been demonstrated to be mutagenic, toxic, pseudo-persistent, bio-accumulative, and to have strong estrogenic effects. Because of their common use in personal care products, they continue to enter the environment. Several food samples, particularly those derived from aquatic sources, have been found to be contaminated with these compounds. Toxic effects on aquatic life, such as metabolic imbalance and developmental toxicity, result from the continued presence of UV absorbers in aquatic bodies. In addition, the degree of exposure to these pollutants in foods should be examined because there are certain risks associated with their consumption by humans. Therefore, this review focuses on the toxicity, analytical techniques, occurrence, and risk assessments of UV absorbers found in food.

Isoeugenol Inhibits Adipogenesis in 3T3-L1 Preadipocytes with Impaired Mitotic Clonal Expansion.

Isoeugenol (IEG), a natural component of clove oil, possesses antioxidant, anti-inflammatory, and antibacterial properties. However, the effects of IEG on adipogenesis have not yet been elucidated. Here, we showed that IEG blocks adipogenesis in 3T3-L1 cells at an early stage. IEG inhibits lipid accumulation in adipocytes in a concentration-dependent manner and reduces the expression of mature adipocyte-related factors including PPARγ, C/EBPα, and FABP4. IEG treatment at different stages of adipogenesis showed that IEG inhibited adipocyte differentiation by suppressing the early stage, as confirmed by lipid accumulation and adipocyte-related biomarkers. The early stage stimulates growth-arrested preadipocytes to enter mitotic clonal expansion (MCE) and initiates their differentiation into adipocytes by regulating cell cycle-related factors. IEG arrested 3T3-L1 preadipocytes in the G0/G1 phase of the cell cycle and attenuated cell cycle-related factors including cyclinD1, CDK6, CDK2, and cyclinB1 during the MCE stage. Furthermore, IEG suppresses reactive oxygen species (ROS) production during MCE and inhibits ROS-related antioxidant enzymes, including superoxide dismutase1 (SOD1) and catalase. The expression of cell proliferation-related biomarkers, including pAKT and pERK1/2, was attenuated by the IEG treatment of 3T3-L1 preadipocytes. These findings suggest that it is a potential therapeutic agent for the treatment of obesity.

Discovery of (-)-epigallocatechin gallate, a novel olfactory receptor 2AT4 agonist that regulates proliferation and apoptosis in leukemia cells.

Olfactory receptors (ORs), the largest family of G protein-coupled receptors (GPCRs), are ectopically expressed in cancer cells and are involved in cellular physiological processes, but their function as anticancer targets is still potential. OR2AT4 is expressed in leukemia cells, influencing the proliferation and apoptosis, yet the limited number of known OR2AT4 agonists makes it challenging to fully generalize the receptor's function. In this study, we aimed to identify new ligands for OR2AT4 and to investigate their functions and mechanisms in K562 leukemia cells. After producing the recombinant OR2AT4 protein, immobilizing it on a surface plasmon resonance chip, and conducting screening to confirm binding activity using 258 chemicals, five novel OR2AT4 ligands were discovered. As a result of examining changes in intracellular calcium by five ligands in OR2AT4-expressing cells and K562 cells, (-)-epigallocatechin gallate (EGCG) was identified as an OR2AT4 agonist in both cells. EGCG reduced the viability of K562 cells and induced apoptosis in K562 cells. EGCG increased the expression of cleaved caspase 3/8 and had no effect on the expression of Bax and Bcl-2, indicating that it induced apoptosis through the extrinsic pathway. Additionally, the initiation of the extrinsic apoptosis pathway in EGCG-induced K562 cells was due to the activation of OR2AT4, using an OR2AT4 antagonist. This study highlights the potential of EGCG as an anti-cancer agent against leukemia and OR2AT4 as a target, making it a new anti-cancer drug.

Obesity exacerbates ischemia-reperfusion injury and senescence in murine kidneys and perirenal adipose tissues.

Background: Obesity is a major worldwide health problem and can be related to cellular senescence. Along with the rise in obesity, the comorbidity of renal ischemia-reperfusion (IR) injury is increasing. Whether obesity accelerates the severity of IR injury and whether senescence contributes to these conditions remain unclear. We studied the degree of injury and cellular senescence in the IR kidneys and perirenal adipose tissues of high-fat-diet-induced obese mice. Methods: C57BL/6 mice fed standard chow or a high-fat diet for 16 weeks were randomized to renal IR or sham group (n = 6-10 each). Renal IR was performed by unilateral clamping of the right renal pedicle for 30 minutes. Six weeks after surgery, renal function, perirenal fat/renal senescence, and histology were evaluated ex vivo. Results: Obese mice showed more renal tubular damage and fibrosis in IR injury than control mice, even though the degree of ischemic insult was comparable. Renal expression of senescence and its secretory phenotype was upregulated in either IR injury or with a high-fat diet and was further increased in the IR kidneys of obese mice. Fat senescence and the expression of tumor necrosis factor alpha were also increased, especially in the perirenal depot of the IR kidneys, with a high-fat diet. Conclusion: A high-fat diet aggravates IR injury in murine kidneys, which is associated, at least in part, with perirenal fat senescence and inflammation. These observations support the exploration of therapeutic targets of the adipo-renal axis in injured obese kidneys.

Monitoring, exposure and risk assessment of formaldehyde in alcoholic beverages in Korea.

Formaldehyde occurs naturally in food and alcoholic beverages. Formaldehyde and alcoholic beverages can cause various health problems, including irritation of the eyes, nose, and throat, respiratory problems, and skin rashes. Alcoholic beverage samples (N = 236) were collected and analyzed for formaldehyde by liquid chromatography-tandem mass spectrometry. The highest average concentrations were detected in fruit wines (1.71 µg/g), followed by wines (1.15 µg/g), cheongju (0.95 µg/g), soju (0.85 µg/g), takju (0.64 µg/g) and beers (0.61 µg/g). We assessed the exposure and risk assessment to formaldehyde from alcoholic beverages based on the monitoring data for the general population and consumers in Korea using various schemes for point estimation. The daily intakes of formaldehyde for the general population and consumers were estimated to be 83 µg and 1202 µg, respectively. The mean hazard indexes (HI) for the general population and consumers in Korea were 0.009 and 0.132, respectively. On the other hand, the mean hazard indexes (HI) for the general population and consumers in Korea were 0.009 and 0.132, respectively. The exposure to formaldehyde in these alcoholic beverages for the Korean population was shown to be of low concern, but it is necessary to monitor the level of formaldehyde in alcoholic beverages and continuously conduct exposure assessment for consumers.

Cinnamyl Alcohol Attenuates Adipogenesis in 3T3-L1 Cells by Arresting the Cell Cycle.

Cinnamyl alcohol (CA) is an aromatic compound found in several plant-based resources and has been shown to exert anti-inflammatory and anti-microbial activities. However, the anti-adipogenic mechanism of CA has not been sufficiently studied. The present study aimed to investigate the effect and mechanism of CA on the regulation of adipogenesis. As evidenced by Oil Red O staining, Western blotting, and real-time PCR (RT-PCR) analyses, CA treatment (6.25-25 µM) for 8 d significantly inhibited lipid accumulation in a concentration-dependent manner and downregulated adipogenesis-related markers (peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein α (C/EBPα), fatty acid binding protein 4 (FABP4), adiponectin, fatty acid synthase (FAS)) in 3-isobutyl-1-methylxanthine, dexamethasone, and insulin(MDI)-treated 3T3-L1 adipocytes. In particular, among the various differentiation stages, the early stage of adipogenesis was critical for the inhibitory effect of CA. Cell cycle analysis using flow cytometry and Western blotting showed that CA effectively inhibited MDI-induced initiation of mitotic clonal expansion (MCE) by arresting the cell cycle in the G0/G1 phase and downregulating the expression of C/EBPß, C/EBPδ, and cell cycle markers (cyclin D1, cyclin-dependent kinase 6 (CDK6), cyclin E1, CDK2, and cyclin B1). Moreover, AMP-activated protein kinase α (AMPKα), acetyl-CoA carboxylase (ACC), and extracellular signal-regulated kinase 1/2 (ERK1/2), markers of upstream signaling pathways, were phosphorylated during MCE by CA. In conclusion, CA can act as an anti-adipogenic agent by inhibiting the AMPKα and ERK1/2 signaling pathways and the cell cycle and may also act as a potential therapeutic agent for obesity.

Pre-treatment with ß-hydroxybutyrate mitigates cisplatin-induced acute kidney injury.

ß-Hydroxybutyrate (ß-HB), the primary circulating ketone body, plays a dual role as both a metabolic fuel and an endogenous signaling molecule, offering diverse systemic benefits. Recent studies have highlighted the renoprotective effects of exogenous ß-HB therapy in various animal models of kidney disease. In this investigation, our goal was to assess whether pre-treatment with exogenous ß-HB could alleviate kidney damage in a mouse model of cisplatin-induced acute kidney injury (AKI). Prior to cisplatin administration, intraperitoneal administration of ß-HB was carried out, and the groups were classified into four: Sham, ß-HB, cisplatin, and ß-HB + cisplatin. The tubular damage score and serum creatinine levels were significantly lower in the ß-HB + cisplatin group compared to the cisplatin group. Furthermore, the expression of phosphorylated NF-κB, inflammatory cytokines, and the quantity of F4/80-positive macrophages in the ß-HB + cisplatin group were reduced compared to those in the cisplatin group. Additionally, oxidative stress markers for DNA, protein, and lipid in the ß-HB + cisplatin group were markedly diminished compared to those in the cisplatin group. The number of TUNEL-positive and cleaved caspase 3-positive tubular cells in the ß-HB + cisplatin group was lower than in the cisplatin group. Pre-treating with exogenous ß-HB effectively mitigated kidney damage by suppressing inflammation, oxidative stress, and tubular apoptosis in cisplatin-induced AKI. Therefore, exogenous ß-HB as a pre-treatment emerges as a promising and novel strategy for preventing cisplatin-induced AKI.

Definitive radiotherapy in patients with clinical T1N0M0 esophageal squamous cell carcinoma: A multicenter retrospective study (KROG 21-10).

PURPOSE: To assess the failure pattern and analyze the treatment scheme of definitive radiation therapy (RT) for T1N0M0 esophageal squamous cell carcinoma (ESCC). METHODS: We performed a multi-institutional retrospective analysis in T1N0M0 ESCC patients who underwent definitive RT from 2010 to 2019. Patterns of failure were demonstrated as in-, and out-field locoregional, and distant metastasis. In the analysis, freedom-from locoregional recurrence (FFLRR) and their association with clinicopathologic factors were evaluated. Propensity score matching in cT1b patients was done. RESULTS: 168 patients were included with a median follow-up of 34.0 months, and 26 cT1a, 116 cT1b disease. The rates of 3-year all and locoregional recurrence for cT1a were 30.5% and 24.1% and those for cT1b were 27.1% and 25.9%, respectively. Among 116 cT1b patients, 69 patients received elective nodal irradiation (ENI) and 47 received involved field irradiation (IFI). After propensity score matching, the 3-year FFLRR rate was 84.5%. There was no difference between ENI and IFI in FFLRR (P = 0.831) and OS (P = 0.525). The 3-year FFLRR was 83.8% (95% Confidence interval (CI), 61.8-93.8%) in IFI group and 85.3% (95% CI, 65.1-94.3%) in ENI group. In multivariate analysis, concurrent chemotherapy use was marginally associated with FFLRR (Hazard ratio, 0.16; P = 0.064). CONCLUSION: cT1a patients who cannot receive endoscopic resection showed similar failure rates as cT1b patients, questioning the staging accuracy and raised the need for thorough treatment like chemoradiotherapy. In cT1b patients, IFI with 50 to 60 Gy and concurrent chemotherapy could be reasonable.

Effect of Polycan, a ß-Glucan from Aureobasidium pullulans SM-2001, on Inflammatory Response and Intestinal Barrier Function in DSS-Induced Ulcerative Colitis.

Ulcerative colitis (UC), a subtype of inflammatory bowel disease, is a chronic gastrointestinal inflammatory disease with unclear etiology and pathophysiology. Herein, we determined the effects of extracellular polysaccharides purified from Aureobasidium pullulans SM-2001 (Polycan) on tight junction protein expression, inflammation, and apoptosis in a dextran sodium sulfate (DSS)-induced acute colitis model. Fifty mice were divided into normal, DSS, DSS + Polycan 250 mg/kg (Polycan 250), DSS + Polycan 500 mg/kg (Polycan 500), and DSS + 5-aminosalicylic acid 100 mg/kg (5-ASA) groups. Their body weights, colon lengths, histological changes in colon tissue, and tight junction function were observed. Results showed that Polycan 250, Polycan 500, and 5-ASA significantly inhibited body weight loss compared with DSS. Similar to 5-ASA, Polycan 500 exhibited preventive effects on colon length shortening and histological changes in colon tissues. Polycan inhibited the DSS-induced decrease in fluorescein isothiocyanate-dextran permeability and myeloperoxidase activity. Moreover, Polycan significantly recovered serum cytokine (e.g., tumor necrosis factor-α, interleukin (IL)-6, and IL-1ß) or mRNA expression in colon tissue compared with DSS. Polycan also inhibited apoptosis by reducing caspase-3 activity and the Bcl-2 associated X/B-cell lymphoma 2 (Bcl-2) ratio. Additionally, DSS treatment significantly reduced microbial abundance and diversity, but the administration of Polycan reversed this effect. Collectively, Polycan protected intestinal barrier function and inhibited inflammation and apoptosis in DSS-induced colitis.

Integrated-Omics Study on the Transcriptomic and Metabolic Changes of Bacillus licheniformis, a Main Microorganism of Fermented Soybeans, According to Alkaline pH and Osmotic Stress.

Bacillus licheniformis has been widely utilized in the food industry as well as various agricultural industries. In particular, it is a main microorganism of fermented soybeans. In this study, the changes of the metabolome and transcriptome of B. licheniformis KACC15844, which had been isolated from fermented soybeans, were investigated depending on alkaline pH (BP) and a high salt concentration (BS) using an integrated-omics technology, focusing on leucine metabolism. Overall, carbohydrate (glycolysis, sugar transport, and overflow) and amino acid (proline, glycine betaine, and serine) metabolisms were strongly associated with BS, while fatty acid metabolism, malate utilization, and branched-chain amino acid-derived volatiles were closely related to BP, in both gene and metabolic expressions. In particular, in leucine metabolism, the formation of 3-methylbutanoic acid, which has strong cheesy odor notes, was markedly increased in BP compared to the other samples. This study provided information on how specific culture conditions can affect gene expressions and metabolite formations in B. licheniformis using an integrated-omics approach.

Simultaneous determination of the free and total forms of nonylphenol, nonylphenol monoethoxylate, and nonylphenol diethoxylate in human urine by gas chromatography-mass spectrometry.

Nonylphenol (NP), nonylphenol monoethoxylate (NP1EO), and nonylphenol diethoxylate (NP2EO) are widely used in various daily products and have been cataloged as endocrine-disrupting chemicals. Due to their high lipophilicity and low biodegradability, these compounds remain in the environment and enter the human body through the food chain. Growing concerns regarding the potential negative effects of NP, NP1EO, and NP2EO on human health have raised the need for biomonitoring to investigate human exposure to these compounds. In this study, a simultaneous analysis method using solid-phase extraction (SPE) combined with gas chromatography-mass spectrometry (GC-MS) was established by controlling the background contamination of NP, NP1EO, and NP2EO, which are ubiquitous in laboratory environments. The proposed method showed proper linearity of over 0.999 and a recovery greater than 85.8% for all analytes. Accuracy and precision were verified in ranges of 92.97-116.30% and 0.65-9.29%, respectively. The detection limits for NP, NP1EO, and NP2EO were 0.0363 µg L-1, 0.0401 µg L-1, and 0.0364 µg L-1, respectively, which were suitable for determining the trace analytes in human urine. Therefore, this simple and integrated analytical method was applied to measure the free and total forms of the target analytes in 25 human urine samples collected in Korea. Overall, free NP, NP1EO, and NP2EO were detected with average contents of 3.94 ± 4.14 µg L-1, 4.63 ± 2.62 µg L-1, and 0.293 ± 0.638 µg L-1, respectively, and with total NP, NP1EO, and NP2EO contents of 6.14 ± 8.24 µg L-1, 5.99 ± 2.91 µg L-1, and 0.806 ± 1.10 µg L-1, respectively. These data showed that these compounds are prevalent in human urine, and indicate the need for further studies.

ß-hydroxybutyrate ameliorates sepsis-induced acute kidney injury.

BACKGROUND: Sepsis is a major cause of acute kidney injury (AKI). Recent studies have demonstrated that ß-hydroxybutyrate (ß-HB) alleviates renal ischemia-reperfusion injury and cisplatin-induced renal injury in murine models. This study aimed to investigate whether ß-HB ameliorates sepsis-induced AKI (SIAKI) in a lipopolysaccharide (LPS)-induced mouse sepsis model. METHODS AND RESULTS: SIAKI was induced by intraperitoneally injecting LPS to C57BL/6 male mice. ß-HB was administrated intraperitoneally before LPS injection. The mice were divided into sham, ß-HB, LPS, and ß-HB + LPS groups. The histological damage score and serum creatinine level were significantly increased in the LPS group mice, but attenuated in the ß-HB + LPS group mice. The expression of phosphorylated nuclear factor-κB tumor necrosis factor-α/interleukin-6 and the number of F4/80-positive macrophages in the ß-HB + LPS group mice were lower than those in the LPS group mice. The number of TdT-mediated dUTP nick-end labeling (TUNEL)-positive tubular cells, cleaved caspase-3 expression, and Bax/Bcl-2 ratio in the ß-HB + LPS group mice were lower than those in the LPS group mice. CONCLUSION: ß-HB pre-treatment ameliorates SIAKI by reducing tubular apoptosis and inflammatory responses. Thus, ß-HB pre-treatment could be a potential prophylactic strategy against SIAKI.

Executive Functions and Morphological Awareness Explain the Shared Variance Between Word Reading and Listening Comprehension.

Purpose: A large body of literature showed that word reading and listening comprehension-two proximal predictors of reading comprehension according to the simple view of reading-are related. Grounded on the direct and indirect effects model of reading (Kim, 2020a, 2020b, 2023), we examined the extent to which the relation is explained by domain-general cognitions or executive functions (working memory and attentional control) and emergent literacy skills (language and code-related skills including morphological awareness, phonological awareness, orthographic pattern recognition, letter naming fluency, and rapid automatized naming). Method: Data were from English-speaking children in Grade 1 (N = 372; 52% boys; 60% White children, 26% African American children, 6% multiracial children, 6% Hispanic children, and 2% Asian American children). Results: Results from structural equation models showed that word reading and listening comprehension were moderately related (.54). When working memory and attentional control were included as predictors, the relation became weaker (.39). When morphological awareness was additionally included, they were no longer related (.05). The other emergent literacy skills did not add explanatory power beyond executive functions and morphological awareness. Conclusion: These results indicate that executive functions and morphological awareness largely explain the shared variance between word reading and listening comprehension for English-speaking beginning readers.

Preparation of Hypoallergenic Whey Protein Hydrolysate by a Mixture of Alcalase and Prozyme and Evaluation of Its Digestibility and Immunoregulatory Properties.

Whey protein (WP) has nutritional value, but the presence of ß-lactoglobulin (ß-LG) and α-lactalbumin (α-LA) cause allergic reactions. In this study, hypoallergenic whey protein hydrolyate (HWPH) was prepared by decomposing ß-LG and α-LA of WP using exo- and endo-type proteases. The enzyme mixing ratio and reaction conditions were optimized using response surface methodology (RSM). Degradation of α-LA and ß-LG was confirmed through gel electrophoresis, and digestion, and absorption rate, and immunostimulatory response were measured using in vitro and in vivo systems. Through RSM analysis, the optimal hydrolysis conditions for degradation of α-LA and ß-LG included a 1:1 mixture of Alcalase and Prozyme reacted for 10 h at a 1.0% enzyme concentration relative to substrate. The molecular weight of HWPH was <5 kDa, and leucine was the prominent free amino acid. Both in vitro and in vivo tests showed that digestibility and intestinal permeability were higher in HWPH than in WP. In BALB/c mice, as compared to WP, HWPH reduced allergic reactions by inducing elevated Type 1/Type 2 helper T cell ratio in the blood, splenocytes, and small intestine. Thus, HWPH may be utilized in a variety of low allergenicity products intended for infants, adults, and the elderly.

Application of Schizosaccharomyces japonicus in makgeolli fermentation and its brewing characteristics.

Recently, unconventional yeasts have become popular as fermentation starters in the brewing industry due to the growing consumer demand for aromatic diversity. Specifically, Schizosaccharomyces japonicus has been explored as a potential starter culture for beer and wine production because of its distinct brewing characteristics; however, its application in makgeolli fermentation has not been tested. Therefore, in the present study, two Sz. japonicus strains (SZJ-1 and SZJ-2) were isolated from natural sources, and their brewing characteristics for makgeolli fermentation were compared with those of commercial S. cerevisiae strain. Although the tested isolates showed a lower fermentation and carbon source consumption rate than control-, their overall alcohol fermentation characteristics were suitable for makgeolli production. Regarding flavor composition, Sz. japonicus-fermented makgeolli possessed more ester compounds (e.g., 2-phenylethyl acetate, ethyl acetate, and ethyl decanoate) than S. cerevisiae-fermented makgeolli. Therefore, Sz. japonicus can be used as an alternative culture starter in makgeolli fermentation. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01265-6.

Disfigured Faces Depicted in Modern Visual Media.

According to the German sociologist Georg Simmel, of all the parts of the body, the face has the highest degree of inner unity. This study aimed to analyze the depictions of disfigured faces in modern visual media (movie films). The keywords "disfigured-face" and "disfigured-face character" were searched on Google. Based on that information, 57 films were searched on IMDB and TMDB, as movie databases. Six films where the IMDB and TMDB overlapped were omitted, and 6 films not directly related to the concept of "disfigured face" were excluded. All animated movies were replaced with the original movies. Then, 45 valid films were finally selected, and 48 characters with disfigured faces were analyzed. Among 48 disfigured faces, 26 were in male characters and 22 were in female characters. Among the 48 disfigured faces, 30 (62.5%) did not improve and 18 (37.5%) improved. Characters who underwent facial deformity correction surgery were 56 times more likely to improve than those who did not. Characters whose facial disfiguration improved were 7.85 times more likely to have a happy ending than those who did not. Among the methods that the characters with disfigured faces used in attempts to overcome their deformity, surgery had the highest success rate. Although surgery is not perfect, the reality is that there are no other alternatives. The results of our film analysis study, which showed plastic surgery can improve the face and improve quality of life, can be seen as realistic and statistically significant.

Oral discourse skills: Dimensionality of comprehension and retell of narrative and expository texts, and the relations of language and cognitive skills to identified dimensions.

We examined the dimensionality of oral discourse skills (comprehension and retell of texts) and the relations of language and cognitive skills to the identified dimensions. Data were from 529 English-speaking second graders (Mage = 7.42; 46% female; 52.6% Whites, 33.8% African Americans, 4.9% Hispanics, 4.7% two or more races, .8% Asian Americans, .6% American Indians, .2% Native Hawaiians, 2.5% unknown; data from 2014-2015 to 2016-2017). Confirmatory factor analysis revealed that oral discourse skills are best described as four related but dissociable dimensions of narrative comprehension, narrative retell, expository comprehension, and expository retell (rs = .59-.84). Language and cognitive skills had different patterns of relations to the identified dimensions and explained larger amounts of variance in comprehension than in retell.

Heukharang lettuce (Lactuca sativa L.) leaf extract displays sleep-promoting effects through GABAA receptor.

ETHNOPHARMACOLOGICAL RELEVANCE: Although lettuce is traditionally known to have hypnotic and sedative effects, to date, only a few studies have documented its sleep-promoting effects and elucidated the related mechanisms. AIM OF THE STUDY: We aimed to investigate the sleep-promoting activity of Heukharang lettuce leaf extract (HLE) with increased lactucin content, known as a sleep-promoting substance in lettuce, in animal models. MATERIALS AND METHODS: To evaluate the effect of HLE on sleep behavior, analysis of electroencephalogram (EEG), gene expression of brain receptors, and activation mechanisms using antagonists were investigated in rodent models. RESULTS: High-performance liquid chromatography analysis showed that HLE contained lactucin (0.78 mg/g of extract) and quercetin-3-glucuronide (1.3 mg/g of extract). In the pentobarbital-induced sleep model, the group administered 150 mg/kg of HLE showed a 47.3% increase in sleep duration time as compared to the normal group (NOR). The EEG analysis showed that the HLE significantly increased non-rapid eye movement (NREM), where delta waves were improved by 59.5% when compared to the NOR, resulting in increased sleep time. In the caffeine-induced arousal model, HLE significantly decreased the awake time increased by caffeine administration (35.5%) and showed a similar level to NOR. In addition, HLE increased the gene and protein expression of gamma-aminobutyric acid receptor type A (GABAA), GABA type B, and 5-hydroxytryptamine (serotonin) receptor 1A. In particular, in comparison to the NOR, the group administered 150 mg/kg HLE showed an increase in expression levels of GABAA and protein by 2.3 and 2.5 times, respectively. When the expression levels were checked using GABAA receptor antagonists, HLE showed similar levels to NOR, as the sleep duration was reduced by flumazenil (45.1%), a benzodiazepine antagonist. CONCLUSIONS: HLE increased NREM sleep and significantly improved sleep behavior due to its action on the GABAA receptors. The collective findings suggest that HLE can be used as a novel sleep-enhancing agent in the pharmaceutical and food industries.

Next-generation sequencing analysis of hepatitis C virus resistance-associated substitutions in direct-acting antiviral failure in South Korea.

BACKGROUND/AIMS: We used next-generation sequencing (NGS) to analyze resistance-associated substitutions (RASs) and retreatment outcomes in patients with chronic hepatitis C virus (HCV) infection who failed direct-acting antiviral agent (DAA) treatment in South Korea. METHODS: Using prospectively collected data from the Korean HCV cohort study, we recruited 36 patients who failed DAA treatment in 10 centers between 2007 and 2020; 29 blood samples were available from 24 patients. RASs were analyzed using NGS. RESULTS: RASs were analyzed for 13 patients with genotype 1b, 10 with genotype 2, and one with genotype 3a. The unsuccessful DAA regimens were daclatasvir+asunaprevir (n=11), sofosbuvir+ribavirin (n=9), ledipasvir/sofosbuvir (n=3), and glecaprevir/pibrentasvir (n=1). In the patients with genotype 1b, NS3, NS5A, and NS5B RASs were detected in eight, seven, and seven of 10 patients at baseline and in four, six, and two of six patients after DAA failure, respectively. Among the 10 patients with genotype 2, the only baseline RAS was NS3 Y56F, which was detected in one patient. NS5A F28C was detected after DAA failure in a patient with genotype 2 infection who was erroneously treated with daclatasvir+asunaprevir. After retreatment, 16 patients had a 100% sustained virological response rate. CONCLUSION: NS3 and NS5A RASs were commonly present at baseline, and there was an increasing trend of NS5A RASs after failed DAA treatment in genotype 1b. However, RASs were rarely present in patients with genotype 2 who were treated with sofosbuvir+ribavirin. Despite baseline or treatment-emergent RASs, retreatment with pan-genotypic DAA was highly successful in Korea, so we encourage active retreatment after unsuccessful DAA treatment.

Comparative Analysis of Volatile and Non-Volatile Metabolites Derived from Bacillus subtilis Strains Producing Different Levels of Biogenic Amines.

Biogenic amines (BAs), which are mainly generated by the microbial decarboxylation of amino acids, are important nitrogen compounds in fermented foods because of their toxicology. However, amino acids, the precursors of BAs, also play an important role in generating volatile and non-volatile metabolites, which are strongly associated with quality indicators for foods. Bacillus subtilis is one of dominant fermentative microorganism in various fermented foods and is well known as a BA-producing bacterium. In this study, B. subtilis strains which have different BAs-producing capacities, higher level of BAs production strain (BH) and lower level of BAs production strain (BL), were applied to compare the formations of volatile and non-volatile metabolite profiles according to cultivation times. In this study, histamine, putrescine, and spermidine were detected in all strains, however, 2-phenylethylamine was detected only in BH. Partial least squares discriminant analysis (PLS-DA) was applied to investigate the difference of metabolic profiles according to strains. In BH, some amino acids (phenylalanine, leucine, and threonine) and related volatile metabolites (3-methylbutanoic acid, pyrazines, styrene, and 1H-indole) were produced higher levels. On the other hand, BL produced significantly higher contents of metabolites associated with metabolism of fatty acids and nucleotides. It is necessary to consider the formation of metabolites in terms of quality as well as that of BAs during fermentation.