Association of Physical Activity With Risk of Liver Fibrosis, Sarcopenia, and Cardiovascular Disease in Nonalcoholic Fatty Liver Disease.

BACKGROUND & AIMS: International guidelines recommend physical activity for subjects with nonalcoholic fatty liver disease (NAFLD). This study investigated the association of physical activity with risk of liver fibrosis, sarcopenia, and cardiovascular disease (CVD) in NAFLD. METHODS: In this multicenter, retrospective study, 11,690 NAFLD subjects who underwent a health screening program and were assessed for physical activity (metabolic equivalent task [MET]-min/week) between 2014 and 2020 were recruited. Liver fibrosis was assessed by using the fibrosis-4 index, NAFLD fibrosis score, and FibroScan-AST score, sarcopenia by using multi-frequency bioelectric impedance analysis, and CVD risk by using atherosclerotic CVD (ASCVD) risk score, and coronary artery calcium (CAC) score were calculated. RESULTS: The prevalence of fibrosis, sarcopenia, high probability of ASCVD, and high CAC score significantly decreased with increasing quartiles of physical activity (all P for trend <.001). In a fully adjusted model, physical activity above 600 MET-min/week (≥third quartile) was independently associated with a reduced risk of fibrosis (adjusted odds ratio [aOR] = 0.59; 95% confidence interval [CI], 0.40-0.86), sarcopenia (aOR = 0.72; 95% CI, 0.58-0.88), high probability of ASCVD (aOR = 0.58; 95% CI, 0.46-0.73), and high CAC score (aOR = 0.32; 95% CI, 0.13-0.83; all P <.05). In addition, increasing amounts of physical activity were significantly associated with risk reduction between fibrosis, sarcopenia, and high probability of ASCVD (all P for trend <.001). In subjects with sarcopenic obesity or lean NAFLD, physical activity was also independently associated with reduced risk of fibrosis and high probability of ASCVD (all P <.05). CONCLUSIONS: Physical activity showed a protective effect against fibrosis, sarcopenia, and CVD in NAFLD.

Spatial clustering of heroin-related overdose incidents: a case study in Cincinnati, Ohio.

BACKGROUND: Drug overdose is one of the top leading causes of accidental death in the U.S., largely due to the opioid epidemic. Although the opioid epidemic is a nationwide issue, it has not affected the nation uniformly. METHODS: We combined multiple data sources, including emergency medical service response, American Community Survey data, and health facilities datasets to analyze distributions of heroin-related overdose incidents in Cincinnati, Ohio at the census block group level. The Ripley's K function and the local Moran's I statistics were performed to examine geographic variation patterns in heroin-related overdose incidents within the study area. Then, conditional cluster maps were plotted to examine a relationship between heroin-related incident rates and sociodemographic characteristics of areas as well as the resources for opioid use disorder treatment. RESULTS: The global spatial analysis indicated that there was a clustered pattern of heroin-related overdose incident rates at every distance across the study area. The univariate local spatial analysis identified 7 hot spot clusters, 27 cold spot clusters, and 1 outlier cluster. Conditional cluster maps showed characteristics of neighborhoods with high heroin overdose rates, such as a higher crime rate, a high percentage of the male, a high poverty level, a lower education level, and a lower income level. The hot spots in the Southwest areas of Cincinnati had longer distances to opioid treatment programs and buprenorphine prescribing physicians than the median, while the hot spots in the South-Central areas of the city had shorter distances to those health resources. CONCLUSIONS: Our study showed that the opioid epidemic disproportionately affected Cincinnati. Multi-phased spatial clustering models based on various data sources can be useful to identify areas that require more policy attention and targeted interventions to alleviate high heroin-related overdose rates.

Anti-Photoaging Effect of Jeju Putgyul (Unripe Citrus) Extracts on Human Dermal Fibroblasts and Ultraviolet B-induced Hairless Mouse Skin.

Ultraviolet (UV) radiation stimulates the expression of matrix metalloproteinases (MMPs) and inflammatory cytokines. These signaling pathways participate in the degradation of the extracellular matrix and induce inflammatory responses that lead to photoaging. This study evaluated the antioxidant activity and the effect on MMPs and procollagen of putgyul extract in vitro. The anti-photoaging activity of putgyul extracts was estimated in vivo using hairless mice (HR-1). The putgyul extracts reduced MMP-1 production and increased the content of procollagen type I carboxy-terminal peptide in human dermal fibroblasts. Ultravilot-B (UVB)-induced expression of inflammatory cytokines and MMPs was detected in mice, and putgyul extracts suppressed the expression. These results suggest that putgyul extract inhibits photoaging by inhibiting the expression of MMPs that degrade collagen and inhibiting cytokines that induce inflammatory responses. The mouse model also demonstrated that oral administration of putgyul extracts decreased wrinkle depth, epidermal thickness, collagen degradation, and trans-epidermal water loss, and increased ß-glucosidase activity on UVB exposed skin. Putgyul extract protects against UVB-induced damage of skin and could be valuable in the prevention of photoaging.

Ulmus macrocarpa Hance Extracts Attenuated H2O2 and UVB-Induced Skin Photo-Aging by Activating Antioxidant Enzymes and Inhibiting MAPK Pathways.

To protect from reactive oxygen species (ROS) damages, skin cells have evolved to have antioxidant enzymes, such as copper and zinc-dependent superoxide dismutase (SOD1), mitochondrial manganese-dependent superoxide dismutase (SOD2), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GR), and suppressed the expression of matrix metalloproteinases (MMPs) through the mitogen-activated protein kinase (MAPK) signaling pathways, such as c-Jun N-terminal kinase (JNK) and p38. Bioactive compounds analyses were performed using a high-performance liquid chromatography-photodiode array detector (HPLC-PDA) system. The antioxidant activity of Ulmus macrocarpa Hance (UMH) extracts was estimated in vitro. The anti-aging activity of UMH extracts was estimated in vivo using the SKH-1 hairless mice. The UMH extracts reduced the H2O2-induced intracellular ROS production and the cell damages in human dermal fibroblasts (HDFs). Moreover, the H2O2-induced phosphorylation of JNK and p38 was detected in HDF and UMH extracts blocked the phosphorylation. These results suggest that UMH extracts can reduce the expression of MMPs and the reduced MMPs lead to the inhibition of collagen degradation. In addition, oral administration of the UMH extracts decreased the depth, thickness, and length of wrinkles on UVB exposed hairless mice. Therefore, UMH extracts play an advantage of the functional materials in antioxidant and anti-aging of skin.

Acute and subchronic (13-week) toxicity of fermented Acanthopanax koreanum extracts in Sprague Dawley rats.

The biological fermentation of plants is usually used to improve their product properties, including their biological activity. Acanthopanax koreanum is a plant indigenous to Jeju, Korea; however, fermented A. koreanum (FAK) has not been guaranteed to be safe. Therefore, in this study, a safety evaluation of aqueous extracts of FAK was performed using Sprague Dawley rats. The acute toxicity of FAK did not influence animal mortality, body weight changes or the animals' clinical appearance at a concentration of 5000 mg/kg body weight. Using doses of 500, 1000 and 2000 mg/kg/day in a subchronic (13-week) toxicity study, the administration of FAK in male rats increased their body weight, food consumption, absolute liver weight, liver-associated enzymes and total cholesterol content. However, these effects of FAK were not considered toxic because the changes were not accompanied by any evidence of clinical signs or any change in the histopathological examination. On the other hand, the FAK-treated female rats did not exhibit significant changes in their body weight, food consumption, absolute and relative organ weights or liver enzymes. These results suggest that the acute oral administration of FAK is non-toxic to rats, and 13 weeks of repeated dosing demonstrated no FAK-related toxicity at a concentration of 2000 mg/kg. Therefore, the no-observed-adverse-effect level (NOAEL) of FAK was determined to be 2000 mg/kg/day for both male and female rats.

Acute and subchronic (28 days) oral toxicity studies of Codonopsis lanceolata extract in Sprague-Dawley rats.

Codonopsis lanceolata is a perennial plant that has been used as a food and in traditional medicine for the treatment of cough, bronchitis, and inflammation in East Asia including Korea, Japan, and China. However, information regarding its toxicity is limited. Therefore, we performed a safety evaluation of aqueous C. lanceolata root extract (CLE) in Sprague-Dawley rats. Assessment of acute toxicity revealed that CLE did not influence mortality, clinical appearance, body weight gain, or necropsy findings at a dose of 5000 mg/kg body weight. In the subchronic oral toxicity, data revealed that several significant alteration in food consumption, water consumption, protein excretion, WBCs levels, TGs, BUN levels, and the absolute and relative weights in the liver, spleen and lungs. However, these changes were transient and were not considered treatment related because they showed no apparent dose dependent. These results suggest that CLE (1250, 2500, and 5000 mg/kg body weight/day) administered orally does not cause acute or subchronic toxicity to male or female rats. The 50% lethal dose (LD50) of CLE was determined to be greater than 5000 mg/kg.

The antioxidant activity and their major antioxidant compounds from Acanthopanax senticosus and A. koreanum.

The antioxidant activity and chlorogenic acid and caffeic acid contents were investigated from different parts of Acanthopanax senticosus and A. koreanum. Antioxidant activity was assessed by various in vitro assays such as DPPH, ABTS, FRAP, reducing power assays and ORAC, and the chlorogenic acid and caffeic acid were validated by HPLC chromatography. Among the various extracts, the fruit extracts of A. senticosus and A. koreanum exhibited strongest antioxidant activities including ABTS, FRAP, reducing power and ORAC, however, strongest DPPH radical scavenging activity was observed from the leaf extract of A. senticosus. In addition, the antioxidant activities of various extracts were correlated with total phenolic and proanthocyanidin contents. The major phenolic contents from various parts of these plants observed that leaf extract of A. senticosus expressed higher levels of chlorogenic acid (14.86 mg/dry weigh g) and caffeic acid (3.09 mg/dry weigh g) than other parts. Therefore, these results suggest that the leaf of A. senticosus may be an excellent natural source for functional foods and pharmaceutical agents, and the validated method was useful for the quality control of A. senticosus.

CoCl2 induces apoptosis through the mitochondria- and death receptor-mediated pathway in the mouse embryonic stem cells.

Embryonic hypoxia/ischemia is a major cause of a poor fetal outcome and future neonatal and adult handicaps. However, biochemical cellular events in mouse embryonic stem (mES) cells during hypoxia remains unclear. This study investigated the underlying mechanism of apoptosis in mES cells under CoCl2-induced hypoxic/ischemic conditions. CoCl2 enhanced the expression of hypoxia-inducible factor-1α (HIF-1α) and the accumulation of reactive oxygen species in mES cells. The CoCl2-treated mES cells showed a decrease in cell viability as well as typical apoptotic changes, cell shrinkage, chromatin condensation, and nuclear fragmentation and an extended G2/M phase of the cell cycle. CoCl2 augmented the release of cytochrome c into the cytosol from the mitochondria with a concomitant loss of the mitochondrial transmembrane potential (ΔΨm) and upregulated the voltage-dependent anion channel. In addition, CoCl2-induced caspase-3, -8, and -9 activation and upregulation of p53 level, whereas downregulated Bcl-2 and Bcl-xL, a member of the anti-apoptotic Bcl-2 family in mES cells. Furthermore, CoCl2 led to the upregulation of Fas and Fas-ligand, which are the death receptor assemblies, as well as the cleavage of Bid in mES cells. These results suggest that CoCl2 induces apoptosis through both mitochondria- and death receptor-mediated pathways that are regulated by the Bcl-2 family in mES cells.

Leucobacter margaritiformis sp. nov., isolated from bamboo extract.

A Gram-positive aerobic rod-shaped non-motile bacterium designated A23(T) was isolated from bamboo extract that had been used to remove odor and was characterized to determine its taxonomic position. 16S rRNA gene sequence analysis revealed that strain A23(T) belongs to the phylum Actinobacteria. The highest degree of sequence similarities was determined to be with Leucobacter salsicius M1-8(T) (96.7%), Leucobacter exalbidus K-540B(T) (96.4%), Leucobacter chromiireducens subsp. chromiireducens L-1(T) (96.4%), Leucobacter komagatae IFO 15245(T) (96.4%) and Leucobacter aerolatus Sj10(T) (96.4%). Chemotaxonomic data revealed that strain A23(T) possesses menaquinone MK11, and its cell wall peptidoglycan contained 2,4-diaminobutyric acid, alanine, glycine, glutamic acid and γ-aminobutyric acid. The polar lipid profile of strain A23(T) contained diphosphatidylglycerol, phosphatidylglycerol and an unknown glycolipid. The predominant fatty acids were iso-C(16:0) (31.5%), anteiso-C(15:0) (43.2%) and anteiso-C(17:0) (13.9%), all of which corroborated the assignment of the strain to the genus Leucobacter. Based on these data, A23(T) (=KEMC 551-022(T) = JCM 17538(T)) should be classified as the type strain for a novel Leucobacter species, for which the name Leucobacter margaritiformis sp. nov. is proposed.

Stabilizing Adversarially Learned One-Class Novelty Detection Using Pseudo Anomalies.

Recently, anomaly scores have been formulated using reconstruction loss of the adversarially learned generators and/or classification loss of discriminators. Unavailability of anomaly examples in the training data makes optimization of such networks challenging. Attributed to the adversarial training, performance of such models fluctuates drastically with each training step, making it difficult to halt the training at an optimal point. In the current study, we propose a robust anomaly detection framework that overcomes such instability by transforming the fundamental role of the discriminator from identifying real vs. fake data to distinguishing good vs. bad quality reconstructions. For this purpose, we propose a method that utilizes the current state as well as an old state of the same generator to create good and bad quality reconstruction examples. The discriminator is trained on these examples to detect the subtle distortions that are often present in the reconstructions of anomalous data. In addition, we propose an efficient generic criterion to stop the training of our model, ensuring elevated performance. Extensive experiments performed on six datasets across multiple domains including image and video based anomaly detection, medical diagnosis, and network security, have demonstrated excellent performance of our approach.

Catalytic reaction mechanism based on alpha-secondary deuterium isotope effects in hydrolysis of trehalose by European honeybee trehalase.

Trehalase, an anomer-inverting glycosidase, hydrolyzes only alpha,alpha-trehalose in natural substrates to release equimolecular beta-glucose and alpha-glucose. Since the hydrolytic reaction is reversible, alpha,alpha-[1,1'-(2)H]trehalose is capable of synthesis from [1-(2)H]glucose through the reverse reaction of trehalase. alpha-Secondary deuterium kinetic isotope effects (alpha-SDKIEs) for the hydrolysis of synthesized alpha,alpha-[1,1'-(2)H]trehalose by honeybee trehalase were measured to examine the catalytic reaction mechanism. Relatively high k(H)/k(D) value of 1.53 for alpha-SDKIEs was observed. The data imply that the catalytic reaction of the trehalase occurs by the oxocarbenium ion intermediate mechanism. In addition, the hydrolytic reaction of glycosidase is discussed from the viewpoint of chemical reactivity for the hydrolysis of acetal in organic chemistry. As to the hydrolytic reaction mechanism of glycosidases, oxocarbenium ion intermediate and nucleophilic displacement mechanisms have been widely recognized, but it is pointed out for the first time that the former mechanism is rational and valid and generally the latter mechanism is unlikely to occur in the hydrolytic reaction of glycosidases.

Synthesis of Polytypic Gallium Phosphide and Gallium Arsenide Nanowires and Their Application as Photodetectors.

One-dimensional semiconductor nanowires often contain polytypic structures, owing to the co-existence of different crystal phases. Therefore, understanding the properties of polytypic structures is of paramount importance for many promising applications in high-performance nanodevices. Herein, we synthesized nanowires of typical III-V semiconductors, namely, gallium phosphide and gallium arsenide by using the chemical vapor transport method. The growth directions ([111] and [211]) could be switched by changing the experimental conditions, such as H2 gas flow; thus, various polytypic structures were produced simultaneously in a controlled manner. The nanobeam electron diffraction technique was employed to obtain strain mapping of the nanowires by visualizing the polytypic structures along the [111] direction. Micro-Raman spectra for individual nanowires were collected, confirming the presence of wurtzite phase in the polytypic nanowires. Further, we fabricated the photodetectors using the single nanowires, and the polytypic structures are shown to decrease the photosensitivity. Our systematic analysis provides important insight into the polytypic structures of nanowires.

Antidiabetic effect of an extract of nutricultured Brassica napus containing vanadium from a Jeju water concentrate.

The purpose of this study was to determine the antidiabetic effect of an extract of nutricultured Brassica napus containing vanadium (BECV). The BECV was prepared following nutriculture of B. napus with a Jeju water vanadium concentrate for 7 day. The BECV was administered to db/db mice for 8 weeks at different dosages (0.028, 0.14, and 0.7 µg/kg; as vanadium concentration in BECV). After 8 weeks, the BECV results showed mouse blood glucose concentrations to significantly decrease, in a dose-dependent manner, compared with the results for control mice. In addition, the concentrations of triglyceride, total cholesterol, and glycated hemoglobin were significantly lower after 8 weeks of administration of 0.7 µg/kg BECV. Therefore, the BECV may have protective effects against type 2 diabetes.

Involvement of both mitochondrial- and death receptor-dependent apoptotic pathways regulated by Bcl-2 family in sodium fluoride-induced apoptosis of the human gingival fibroblasts.

Sodium fluoride (NaF) has been shown to be cytotoxic and produces inflammatory responses in humans. However, the cellular mechanisms underlying the NaF-induced cytotoxicity in periodontal tissues are unclear. This study examined whether or not NaF induces apoptosis in human gingival fibroblasts (HGF), and its underlying mechanisms by monitoring various apoptosis-associated processes. NaF reduced the cell viability of HGF in a dose- and time-dependent manner. NaF increased TUNEL-positive cell and induced apoptosis with concomitant chromatin condensation and DNA fragmentation in HGF. In addition, NaF increased the level of cytochrome c released from the mitochondria into the cytosol, enhanced the caspase-9, -8 and -3 activities, the cleavage of poly (ADP-ribose) polymerase (PARP), and up-regulated the voltage-dependent anion channel (VDAC) 1. However, NaF did not affect the production of reactive oxygen species (ROS) which is a strong apoptotic inducer. Furthermore, NaF up-regulated the Fas-ligand (Fas-L), a ligand of death receptor. Bcl-2, a member of the anti-apoptotic Bcl-2 family, was down-regulated, whereas the expression of Bax, a member of the pro-apoptotic Bcl-2 family, was unaffected in the NaF-treated HGF. These results suggest that NaF induces apoptosis in HGF through both the mitochondria-mediated pathways regulated by the Bcl-2 family and death receptor-mediated pathway.

Strain Mapping and Raman Spectroscopy of Bent GaP and GaAs Nanowires.

Strain engineering of nanowires (NWs) has been recognized as a powerful strategy for tuning the optical and electronic properties of nanoscale semiconductors. Therefore, the characterization of the strains with nanometer-scale spatial resolution is of great importance for various promising applications. In the present work, we synthesized single-crystalline zinc blende phase GaP and GaAs NWs using the chemical vapor transport method and visualized their bending strains (up to 3%) with high precision using the nanobeam electron diffraction technique. The strain mapping at all crystallographic axes revealed that (i) maximum strain exists along the growth direction ([111]) with the tensile and compressive strains at the outer and inner parts, respectively; (ii) the opposite strains appeared along the perpendicular direction ([2̅11]); and (iii) the tensile strain was larger than the coexisting compressive strain at all axes. The Raman spectrum collected for individual bent NWs showed the peak broadening and red shift of the transverse optical modes that were well-correlated with the strain maps. These results are consistent with the larger mechanical modulus of GaP than that of GaAs. Our work provides new insight into the bending strain of III-V semiconductors, which is of paramount importance in the performance of flexible or bendable electronics.

Investigating a Needle-Based Epidural Procedure in Obstetric Anesthesia.

This study investigates the safety and efficacy of a large-dose, needle-based epidural technique in obstetric anesthesia. The technique differs from a standard, catheter-based approach in that the anesthetic dose is administered through an epidural needle prior to insertion of the epidural catheter. Using a data-driven informatics and machine learning approach, our findings show that the needle-based technique is faster and more dose-effective in achieving sensory level. We also find that injecting large doses in the epidural space through the epidural needle is safe, with complication rates similar to those reported in published literature for catheter-based technique. Further, machine learning reveals that if the needle dose is kept under 18 ml, the resulting hypotension rate will be significantly lower than published results. The machine learning framework can predict the incidence of hypotension with 85% accuracy. The findings from this investigation facilitate delivery improvement and establish an improved clinical practice guideline for training and for dissemination of safe practice.

Protective effects of citrus based mixture drinks (CBMDs) on oxidative stress and restraint stress.

In the current study investigated the protective effects of citrus based mixture drinks (CBMDs) using oxidative stress in human dermal fibroblast (HDF) cells and restraint-stressed rats. The CBMDs contained citrus bioflavonoids including narirutin and hesperidin. The cell viability of HDF cells treated with H2O2 was observed at 53.9% but treated with CBMD-1 and CBMD-2 (500 µg/mL) on H2O2 exposed HDF cells significantly increased the relative cell viability at 65.0 and 72.2%, respectively. In the treadmill test, the time spent on the electrode plate in the restraint-stressed group was analyzed 24.1 s, but restraint-stressed rats with administered CBMDs (300 mg/kg) had significantly decreased the time at 2.4 (CBMD-1) and 4.7 (CBMD-2) s, respectively. In addition, number of touches the electrode plate in restraint-stressed group was observed at 42.4 ea, but, restraint-stressed rats with administered CBMD-1 and CBMD-2 (300 mg/kg) were significantly decreased at 7.0 and 10.2 ea, respectively.

Toxicological Evaluation of Brassica napus Extract Containing Vanadium, Nutricultured in Jeju Water.

This study evaluated the mutagenicity and acute toxicity of the juice extract of nutricultured Brassica napus containing vanadium (BECV). The BECV was prepared by nutriculture for 7 days in Jeju water containing vanadium. The mutagenic effects of BECV were investigated using the bacterial reverse mutation test, chromosome aberration test, and micronucleus test. Based on the results of the mutagenicity test, we propose that BECV is not a mutagenicity-inducing agent. In the acute oral toxicity study, male and female Sprague-Dawley rats were administered a single limiting dose of 0.014, 0.14, or 1.4 µg BECV/kg body weight; the rats were then observed for 7 days. No acute lethal effect was observed at the maximal dose of 1.4 µg BECV/kg body weight. In the subacute study, male and female rats were administered once daily, by oral gavage, a dose of 0.028, 0.14, and 0.7 µg/kg body weight of BECV for 28 days. No significant toxicity was observed not only hematological, biochemical, and pathological parameters but also the body and organ weights when compared to controls. The level of BECV with no observed adverse effects in male and female rats was 0.7 µg/kg body weight (concentration of vanadium in BECV) in the subacute toxicity study.

Radical scavenging-linked anti-adipogenic activity of Alnus firma extracts.

The purpose of the present study was to investigate the antioxidant activity and anti-adipogenic effect of extracts from Alnus firma (A. firma), which is an edible plant that grows in mountainous areas. The total phenolic, flavonoid and anthocyanin content as well as the antioxidant activity of a 70% ethanolic extract of A. firma (AFE) was assessed. Furthermore, the effects of AFE on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis of 3T3-L1 cells were investigated. The results revealed that the total phenolic, flavonoid and pro-anthocyanidin content of AFE as 436.26±3.30 mg gallic acid equivalents/g, 73.82±0.54 mg quercetin equivalents/g and 149.25±6.06 mg catechin equivalents/g, respectively. In addition, AFE exerted significant antioxidant effects in terms of 1,1-diphenyl-2-picryl hydrazyl radical scavenging activity, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity, reducing power, oxygen radical absorbance capacity and nitric oxide radical scavenging activity. As for its anti-adipogenic activity, AFE significantly inhibited ROS production and lipid accumulation during adipogenesis of 3T3-L1 cells compared with those in control cells. In addition, AFE regulated adipogenic transcription factors including peroxisome proliferator­activated receptor-γ, CCAAT/enhance-binding protein α and adipocyte protein 2. These results indicated that A. firma is a potential candidate for a functional food supplement.

Identifying method validation and measurement uncertainty of brominated vegetable oil in soft drinks and carbonated waters commonly consumed in South Korea.

This study investigated a method for validating and determining the measurement uncertainty for the composition of brominated vegetable oil (BVO) in soft drinks and carbonated waters commonly consumed in South Korea. First, we studied a simple and precise qualitative colorimetric method at the maximum residues level 15 ppm. And an analytical method using ion chromatography (IC) was validated and identified with brominated fatty acids by gas chromatography electron ionization mass spectrometry (GC/EI-MS). The measurement uncertainty was evaluated based on the precisional study and confirmed by the preliminary inter-laboratory study. For IC analysis, the recovery range of BVO was from 97.8% to 107.2% with relative standard deviations between 0.18% and 0.69%. In addition, the expanded uncertainty of the BVO was 1.59. These results indicate that the validated method is appropriate for identifying of BVO and can be used to verify the safety of soft drinks or carbonated waters containing BVO residues.