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Accidental chemical leaks and illegal chemical discharges are a global environmental issue. In 2012, a hydrogen fluoride leak in Gumi, South Korea, killed several people and contaminated the environment. This leak also led to a significant decline in crop yield, even after the soil concentration of hydrogen fluoride decreased to below the standard level following natural attenuation. To determine the cause of this decreased plant productivity, we designed direct and indirect exposure tests by evaluating the metabolome, transcriptome, and phenome of the plants. In an indirect exposure test, soil metabolomics revealed downregulation of metabolites in vitamin B6, lipopolysaccharide, osmolyte, and exopolysaccharide metabolism. Next-generation sequencing of the plants showed that ABR1 and DREB1A were overexpressed in response to stress. Plant metabolomics demonstrated upregulation of folate biosynthesis and nicotinate and nicotinamide metabolism associated with detoxification of reactive oxygen species. These results demonstrate impaired metabolism of soil microbes and plants even after natural attenuation of hydrogen fluoride in soil. The novel chemical exposure testing used in this study can be applied to identify hidden damage to organisms after natural attenuation of chemicals in soil, as well as biomarkers for explaining the decline in yield of plants grown in soil near pollutant-emitting industrial facilities.

Metabolites reflect the biochemical dynamics for the maintenance of pregnancy and parturition. UPLC-Q/TOF-MS and LC-MS/MS metabolomics were performed to identify and validate the plasma metabolomic signatures of preterm birth (PTB). We recruited pregnant women between 16 and 40 weeks 5 days gestational age at Ewha Womans Mokdong Hospital for a nested case-control study. In untargeted UPLC-Q/TOF-MS, score plots of partial least-squares discriminant analysis clearly separated the PTB group from the term birth (TB, n = 10, PTB, n = 11). Fifteen metabolites were significantly different between the two groups, as indicated by a variable importance in projection &gt, 1 and p &lt, 0.05. Metabolic pathways involving retinol, linoleic acid, D-arginine, and D-ornithine were associated with PTB. Verification by LC-MS/MS focused on retinol metabolism (TB, n = 39, PTB, n = 20). Retinol levels were significantly reduced in PTB compared to TB, while retinal palmitate, all-trans-retinal, and 13-cis-retinoic acid (13cis-RA) significantly increased (p &lt, 0.05). Retinol-binding protein levels were also elevated in PTB. Additionally, all-trans-retinal (AUC 0.808, 95% CI: 0.683–0.933) and 13cis-RA (AUC 0.826, 95% CI: 0.723–0.930) showed improved predictions for PTB-related retinol metabolites. This study suggests that retinoid metabolism improves the accuracy of PTB predictions and plays an important role in maintaining pregnancy and inducing early parturition.

Hair follicles are among the most highly proliferative tissues. Polyamines are associated with proliferation, and several polyamines including spermidine and spermine play anti-inflammatory roles. Androgenic alopecia results from increased dihydrotestosterone metabolism, and alopecia areata is an autoimmune disease. This study aimed to investigate differences in polyamine profiles in hair samples between patients with androgenic alopecia and alopecia areata. Polyamine concentrations were determined through high-performance liquid chromatography-mass spectrometry. Hair samples were derivatized with isobutyl chloroformate. Differences in polyamine levels were observed between androgenic alopecia and alopecia areata compared with normal controls. In particular, polyamine levels were higher in alopecia areata patients than in normal controls. Certain polyamines displayed different concentrations between the androgenic alopecia and alopecia areata groups, suggesting that some polyamines, particularly N-acetyl putrescine (P = 0.007) and N-acetyl cadaverine (P = 0.0021), are significantly different in androgenic alopecia. Furthermore, spermidine (P = 0.021) was significantly different in alopecia areata. Our findings suggest that non-invasive quantification of hair polyamines may help distinguish between androgenic alopecia and alopecia areata. Our study provides novel insights into physiological alterations in patients with androgenic alopecia and those with alopecia areata and reveals some differences in polyamine levels in hair loss diseases with two different modes of action.

Hair loss, from the vertex or front of the head, generally occurs due to increased androgenic steroid levels. Androgenic steroids, particularly testosterone and dihydrotestosterone, are distributed differently across the vertex and occipital regions and are involved in inducing ornithine decarboxylase expression. Therefore, we hypothesized that the distribution of polyamines may be altered in different scalp regions. For the overall metabolic profiling of polyamines in patients with hair loss, a liquid chromatography-mass spectrometry was used. We investigated the differential polyamine levels in different regions of the hair of patients with male pattern baldness and those with female pattern hair loss. The levels of most polyamines were higher in the vertex region than in the occipital region, and N-acetyl polyamine levels differed significantly. We proposed to test our hypothesis by profiling polyamines in human hair fibre to evaluate the distribution of metabolites in various regions of the scalp.

Despite significant advances in neuroscience research over the past several decades, the exact cause of AD has not yet fully understood. The metabolic hypothesis as well as the amyloid and tau hypotheses have been proposed to be associated with AD pathogenesis. In order to identify metabolome signatures from the postmortem brains of sporadic AD patients and control subjects, we performed ultra performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometer (UPLC-LTQ–Orbitrap-MS). Not only our study identified new metabolome signatures but also verified previously known metabolome profiles in the brain. Statistical modeling of the analytical data and validation of the structural assignments discovered metabolic biomarkers associated with the AD pathogenesis. Interestingly, hypotaurin, myo-inositol and oxo-proline levels were markedly elevated in AD while lutamate and N-acetyl-aspartate were decreased in the postmortem brain tissue of AD patients. In addition, neurosteroid level such as cortisol was significantly increased in AD. Together, our data indicate that impaired amino acid metabolism is associated with AD pathogenesis and the altered amino acid signatures can be useful diagnostic biomarkers of AD. Thus, modulation of amino acid metabolism may be a possible therapeutic approach to treat AD., Graphical Abstract

A simultaneous quantitative profiling method for androgens and prostaglandins using ultra-high-performance liquid chromatography–tandem mass spectrometry was developed and validated to evaluate urinary androgen and prostaglandin levels. Solid-phase extraction and liquid–liquid extraction steps were combined during the sample preparation. β-Glucuronidase/arylsulfatase was also used in the enzyme hydrolysis step. Chemical derivatization was performed using 2-hydrazinopyridine for simultaneous determination of androgen and prostaglandin in the same ionization mode. The analytes were all separated and measured using multiple reaction monitoring in the positive ion mode within a run time of 22 min. The method was validated, achieving overall recoveries ranging from 81.0 to 102.9% with limits of quantification ranging from 0.01 to 2 ng/mL. The intra-day accuracy and precision ranged from 6.5 to 14.3% and from 77.1 to 106.8%, respectively. The inter-day accuracy and precision ranged from 8.9 to 18.2% and 89.9 to 101.4%, respectively. The linearity was expressed using the correlation coefficient, which was >0.99. The method developed herein was used to investigate the effects of a one-year finasteride treatment through differences in urinary androgen and prostaglandin levels between treated male pattern baldness patients and normal controls. The urinary androgen and prostaglandin levels were not significantly different between the two groups because of the administration of finasteride. The results confirmed that finasteride affects androgens and PGs related to hair regrowth and growth length, and a one-year finasteride treatment is effective for MPB. The mass spectrometry-based quantitative profiling method used herein for the investigation of male pattern baldness also holds great potential for the evaluation of androgens and prostaglandins associated with the metabolism of various inflammatory diseases.

Chemical accidents have frequently occurred in South Korea as a result of the huge amount of chemicals being used in various industries. Even though fire accidents accounted for 71.9% of chemical accidents during 2008–2018 in South Korea, most ecological research and investigation has focused on leakage accidents since most fire or explosion gases are diffused out and disappear into the atmosphere. In this study, the possibility of soil contamination by toluene combustion is proposed. A fire simulation batch test was performed and identified the combustion by-products such as methylbenzene, ethylbenzene, ethynylbenzene, benzaldehyde, 1-phenyl-1-propyne, naphthalene, 2-methylindene using gas chromatography coupled with mass spectrometry (GC–MS). Naphthyl-2-methyl-succinic acid, a metabolic intermediate of naphthalene metabolism derived from the combustion product of toluene, was also discovered in field soil and the secondary metabolites such as streptomycin 6-phosphate, 3-Nitroacrylate, oxaloacetate using LC–MS. Moreover, Streptomyces scabiei, participating in naphthalene metabolism, was also discovered in filed soil (contaminated soil) using 16s rRNA sequencing. As a result, bacterial stress responses in field soil (contaminated soil) affected by gas cloud were identified by discovering metabolites relating to bacterial self-defense action such as fatty biosynthesis. This study draws a conclusion that soil can be polluted enough to affect bacteria by gas cloud and soil bacteria and can encounter stress for a long term even though toluene and its combustion products had already decomposed in soil.

This study investigated the effects of accidental contamination of soils with phenol, toluene, nitric acid, and hydrogen fluoride (HF) by simulating chemical leakage in the soil with/without rain and characterizing the resulting metabolites and microbial. In the case of acid leakage, pH and cation exchange capacity were decreased, and the content of fluoride ion was increased in case of HF leakage. Using mass spectrometry-based metabolomics analysis, phytosphingosine was detected as a distinguishing metabolite in soils contaminated with phenol and HF in rain conditions. Microbial communities were identified by 16s rRNA metagenome sequencing. Sphingomonas was one of the dominant species in soils contaminated with phenol and HF. These results suggest that phytosphingosine and Sphingomonas might be used as biomarkers to evaluate the status of soils contaminated with phenol or HF. Under simulated rain conditions, the species alpha-diversity index of soil microbes and the physicochemical properties of the soil indicated values close to those of the uncontaminated soil. Rain played an important role in the recovery of microbial and metabolic profiles after chemical accidents. Metabolic profiling and microbial community analysis can serve as a diagnostic tool for ecotoxicological research at chemical accident sites.

Phthalate exposure has been reported to be more associated with obesity in children than in adults. The concentration of di(2-ethylhexyl)phthalate (DEHP) was high temporal variability in spot urine, so additional tools of assessing DEHP exposure were required. Therefore, we used relative metabolite ratios (RMRs) as well as concentrations, and RMRs did not need to be corrected to the creatinine concentration. We aimed to evaluate the levels of urinary DEHP metabolites and their RMRs in obese children in South Korea, and to investigate the potential of RMRs for assessing the risks for childhood obesity. We analyzed the four urinary DEHP metabolites (mono-2-ethylhexyl phthalate (MEHP), mono-(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono-(2-ethyl-5-oxohexyl) phthalate (MEOHP), and mono-(2-ethyl-5-carboxypentyl) phthalate (MECPP)) in 240 children aged 5–16 years, using isotope dilution GC-MS/MS. The children were placed into three groups (“normal weight,” “overweight,” and “obese”) according to body mass index (BMI) percentiles. We statistically compared the concentrations and RMRs of DEHP metabolites among these groups. The obese group had lower MEHP levels, and higher secondary metabolite (MEHHP, MEOHP, and MECPP) levels, than the normal weight group. DEHP metabolite levels did not differ significantly between the normal weight and obese groups, whereas RMRA2 (as the ratio of the molar concentrations of MEOHP to MEHHP) was found to be negatively associated with BMI percentile (β= −0.236, p

Male pattern baldness (MPB) has been associated with dihydrotestosterone (DHT) expression. Finasteride treats MPB by inhibiting 5-alpha reductase and blocking DHT production. In this study, we aimed to identify metabolic differences in urinary metabolomics profiles between MPB patients after a one-year treatment with finasteride and healthy controls. Untargeted and targeted metabolomics profiling was performed using liquid chromatography-mass spectrometry (LC-MS). We hypothesized that there would be changes in overall metabolite concentrations, especially steroids, in the urine of hair loss patients treated with finasteride and normal subjects. Untargeted analysis indicated differences in steroid hormone biosynthesis. Therefore, we conducted targeted profiling for steroid hormone biosynthesis to identify potential biomarkers, especially androgens and estrogens. Our study confirmed the differences in the concentration of urinary androgens and estrogens between healthy controls and MPB patients. Moreover, the effect of finasteride was confirmed by the DHT/T ratio in urine samples of MPB patients. Our metabolomics approach provided insight into the physiological alterations in MPB patients who have been treated with finasteride for a year and provided evidence for the association of finasteride and estrogen levels. Through a targeted approach, our results suggest that urinary estrogens must be studied in relation to MPB and post-finasteride syndrome.

We developed a colorimetric method for measuring the amount of oil in mouse stool after co-administering an oil-soluble dye. When the amount of oil in the feces calculated from the amounts of Sudan III and Oil Red O was plotted against the amount of oil detected by liquid chromatography-mass spectrometry, the graph was linear, showing a one-to-one correlation between two analyses. This method may be utilized to determine the efficacy of lipase inhibitors, or to assess fat malabsorption in vivo.

This study aimed to measure phytoestrogen contents in health functional foods (HFFs) to alleviate menopausal symptoms that are commercially available in South Korea. A sensitive and selective method using gas chromatography–mass spectrometry (GC-MS) for the monitoring of 21 phytoestrogens was developed and fully validated. Phytoestrogens were extracted by liquid–liquid extraction using methyl tert-butylether as the solvent. Food samples were enzymatically hydrolyzed with β-glucuronidase/arylsulfatase. Before GC-MS, the analytes were derivatized to their trimethylsilyl derivatives by incubation with N,O-bis(trimethylsilyl)trifluoroacetamide with 1% trimethylchlorosilane for 30 min at 80°C. The calibration curves showed good linearity (r 2 > 0.99) in the range of interest for each analyte. The limits of detection and quantification of the 21 phytoestrogens were in the ranges of 0.5–20 and 1–50 ng/mL, respectively. The intra- and inter-day precisions were all below 15.7%. These multivariate procedures were efficient in determining the optimal conditions using peak heights as responses.

Alopecia areata is a well-known autoimmune disease affecting humans. Polyamines are closely associated with proliferation and inflammation, and steroid hormones are involved in immune responses. Additionally, bile acids play roles in immune homeostasis by activating various signaling pathways; however, the roles of these substances and their metabolites in alopecia areata remain unclear. In this study, we aimed to identify differences in metabolite levels in urine samples from patients with alopecia areata and healthy controls. To assess polyamine, androgen, and bile acid concentrations, we performed high-performance liquid chromatography–tandem mass spectrometry. Our results showed that spermine and dehydroepiandrosterone levels differed significantly between male patients and controls, whereas ursodeoxycholic acid levels were significantly higher in female patients with alopecia areata than in controls. Our findings suggested different urinary polyamine, androgen, and bile acid concentrations between alopecia areata patients and normal controls. Additionally, levels of endogenous substances varied according to sex, and this should be considered when developing appropriate treatments and diagnostic techniques. Our findings improve our understanding of polyamine, androgen, and bile acid profiles in patients with alopecia areata and highlight the need to consider sex-related differences.

Fetal programming implies that the maternal diet during pregnancy affects the long-term health of offspring. Although maternal diet influences metabolic disorders and non-alcoholic fatty liver disease in offspring, the hepatic mechanisms related to metabolites are still unknown. Here, we investigated the maternal diet-related alterations in metabolites and the biological pathway in male offspring at three months of age. Pregnant rats were exposed to 50% food restriction during the prenatal period or a 45% high-fat diet during the prenatal and postnatal periods. The male offspring exposed to food restriction and high-fat diets had lower birth weights than controls, but had a catch-up growth spurt at three months of age. Hepatic taurine levels decreased in both groups compared to controls. The decreased hepatic taurine levels in offspring affected excessive lipid accumulation through changes in hepatocyte nuclear factor 4 A methylation. Moreover, the alteration of gluconeogenesis in offspring exposed to food restriction was observed to a similar extent as that of offspring exposed to a high fat diet. These results indicate that maternal diet affects the dysregulation in hepatic metabolism through changes in taurine levels and HNF4A methylation, and predisposes the offspring to Type 2 diabetes and non-alcoholic fatty liver disease in later life.

Trihalomethanes (THMs) are chemicals regulated by Environmental Protection Agency's first drinking water regulation issued after the passage of the Safe Drinking Water Act. Among THMs, iodotrihalomethanes (I-THMs) are produced by treating water containing iodides ion with chlorine or ozone. I-THMs are more carcinogenic and biotoxic than chlorinated or brominated THMs. The purpose of this study was to analyze of I-THMs in drinking water using the liquid-liquid extraction (LLE) method with various extraction solvents. The calibration curves ranged from 0.01 to 20 ng/mL and the correlation coefficient showed a good linearity of 0.99 or more. The method detection limit ranged from 0.01 to 0.10 ng/mL. The accuracy of the LLE method ranged from 99.43 to 112.40%, and its precision ranged from 1.10 to 10.36%. Good recoveries (71.35-118.60%) were obtained for spiked drinking water samples, demonstrating that the LLE method is suitable for the analysis of drinking water samples. Dichloroiodomethane, bromochloroiodomethane, and dibromoiodomethane were identified in drinking water collected from 70 places of water purification plants in Korea. The samples were classified by disinfection systems, regions, seasons, and water sources. The concentration of I-THMs in pre-/postchlorination facilities owing to excess chlorine usage was higher than in ozonization/postchlorination facilities. Moreover, the concentrations of I-THMs were high in the coastal region, because of the large amount of halide ions from the sea. There was no seasonal difference; however, the concentration of I-THMs in pre-/postchlorination facilities increased in spring and summer. The concentration of I-THMs in water sources was high in samples from the Geum River and the Yeongsan and Sumjin River. The concentration and detection frequency of I-THMs in Han River and Nakdong River were high in the coastal region, because of numerous pre-/postchlorination facilities and the abundance of halide ions from the ocean.

Purpose Beyond the limited scope of non-specific polyclonal regulatory T cell (Treg)-based immunotherapy, which depends largely on serendipity, the present study explored a target Treg subset appropriate for the delivery of a novel epitope spreader Pep19 antigen as part of a sophisticated form of immunotherapy with defined antigen specificity that induces immune tolerance. Methods Human polyclonal CD4+CD25+CD127lo− Tregs (127-Tregs) and naïve CD4+CD25+CD45RA+ Tregs (45RA-Tregs) were isolated and were stimulated with target peptide 19 (Pep19)-pulsed dendritic cells in a tolerogenic milieu followed by ex vivo expansion. Low-dose interleukin-2 (IL-2) and rapamycin were added to selectively exclude the outgrowth of contaminating effector T cells (Teffs). The following parameters were investigated in the expanded antigen-specific Tregs: the distinct expression of the immunosuppressive Treg marker Foxp3, epigenetic stability (demethylation in the Treg-specific demethylated region), the suppression of Teffs, expression of the homing receptors CD62L/CCR7, and CD95L-mediated apoptosis. The expanded Tregs were adoptively transferred into an NOD/scid/IL-2Rγ−/− mouse model of collagen-induced arthritis. Results Epitope-spreader Pep19 targeting by 45RA-Tregs led to an outstanding in vitro suppressive T cell fate characterized by robust ex vivo expansion, the salient expression of Foxp3, high epigenetic stability, enhanced T cell suppression, modest expression of CD62L/CCR7, and higher resistance to CD95L-mediated apoptosis. After adoptive transfer, the distinct fate of these T cells demonstrated a potent in vivo immunotherapeutic capability, as indicated by the complete elimination of footpad swelling, prolonged survival, minimal histopathological changes, and preferential localization of CD4+CD25+ Tregs at the articular joints in a mechanistic and orchestrated way. Conclusions We propose human naïve CD4+CD25+CD45RA+ Tregs and the epitope spreader Pep19 as cellular and molecular targets for a novel antigen-specific Treg-based vaccination against collagen-induced arthritis., Graphical Abstract

Tryptophan (Trp) is an essential amino acid that plays an important role in protein synthesis and is a precursor of various substances related to diverse biological functions. An imbalance in Trp metabolites is associated with inflammatory diseases. The accurate and precise measurement of these compounds in biological specimens would provide meaningful information for understanding the biochemical states of various metabolic syndrome-related diseases, such as hyperlipidemia, hypertension, diabetes, and obesity. In this study, we developed a rapid, accurate, and sensitive liquid chromatography-tandem mass spectrometry-based method for the simultaneous targeted analysis of Trp and its related metabolites of the kynurenine (Kyn), serotonin, and tryptamine pathways in urine. The application of the developed method was tested using urine samples after protein precipitation. The detection limits of Trp and its metabolites were in the range of 0.01 to 0.1 μg/mL. The method was successfully validated and applied to urine samples from controls and patients with metabolic syndrome. Our results revealed high concentrations of Kyn, kynurenic acid, xanthurenic acid, and quinolinic acid as well as a high Kyn-to-Trp ratio (KTR) in patients with metabolic syndromes. The levels of urine Kyn and KTR were significantly increased in patients under 60 years old. The profiling of urinary Trp metabolites could be a useful indicator for age-related diseases including metabolic syndrome. ᅟ.

Alkoxyalcohols are used as solvents or preservatives in various consumer products such as wet wipes. The metabolites of alkoxyalcohols are known to be chronically toxic and carcinogenic to animals. Thus, an analytical method is needed to monitor alkoxyalcohols in wet wipes. The aim of this study was to develop a simultaneous analytical method for 14 alkoxyalcohols using headspace gas chromatography coupled with mass spectrometry to analyze the wet wipes. This method was developed by comparing with various headspace extraction parameters. The linear calibration curves were obtained for the method (r 2 > 0.995). The limit of detection of alkoxyalcohols ranged from 2 to 200 ng mL �1 . The precision of the determinative method was less than 18.20% coefficient of variation both intra and inter days. The accuracy of the method ranged from 82.86% to 119.83%. (2-Methoxymethylethoxy)propanol, 2-phenoxyethanol, and 1-phenoxy-2-propanol were mainly detected in wet wipes.

Phthalates are used in industry products, household items, and medical tools as plasticizers. Human exposure to phthalates has raised concern about its toxicity. In the present study, optimization was conducted for the simultaneous analysis of eight kinds of phthalate metabolites using gas chromatography-mass spectrometry (GC-MS): MEP, MiBP, MnBP, MBzP, MiNP, MEHP, MEOHP, and MEHHP. In order to minimize the matrix effect and to do quantitative analysis, isotope dilution and LLE-GC-MS methods were performed. Urine samples were enzymatically hydrolyzed, extracted with a mixture of n-hexane and ethyl ether (8:2; v:v), and subsequently derivatized with trimethylsilylation. All eight kinds of analytes showed clear resolution and high reproducibility in GC-MS results. The method detection limit ranged from 0.05 ng/mL to 0.2 ng/mL. Calibration curves were found to be linear from 0.2 to 100 ng/mL with -(2)>0.992. The relative standard deviation of the intraday precision using water and urine ranged from 2.1% to 16.3%. The analysis was performed with urine samples that were collected from adults residing in the Republic of Korea. The analyzed concentration results were compared according to gender and region. As a result, DEHP metabolites showed the highest detected concentration (75.92 μg/g creatinine, 100%), and MiNP, a metabolite of DiNP, showed the lowest detected concentration (0.42 μg/g creatinine, 22.5%). On average, female urine (200.76 μg/g creatinine) had a higher detected concentration of ∑8 phthalate metabolites than male urine. Samples from rural regions (211.96 μg/g creatinine) had higher levels than samples from urban regions.

The authors found a language error in the published article. The authors replace the Figure 1.

PURPOSE: The purpose of this study was to investigate the effects of thermotherapy on gas pain, post-operative resilience, and body temperature discomfort among patients who received laparoscopic myomectomies. METHODS: The experimental group consisted of 62 patients with thermotherapy and the control group consisted of 60 patients. Thermotherapy was applied individually to the experimental group four hours after surgery. The collected data was analyzed using descriptive statistics, t-tests, χ²-tests, and repeated measures of analysis of variance, using IBM SPSS Statistics version 18. RESULTS: The results showed no significant interaction effect between the group and time of measurement in gas-related pain in the experimental group. For gas-related pain, there was significant difference in right shoulder pain at 24 hours (t=-4.222, p=.000), 48 hours (t=-3.688, p=.000), 72 hours (t=-2.250, p=.028), and left at 24 hours (t=-3.727, p=.000), 48 hours (t=-4.150, p=.000), and 72 hours (t=-2.482, p=.016) and both shoulders at 24 hours (t=-2.722, p=.009) and 48 hours (t=-2.525, p=.014). There was no significant difference in epigastric pain, excluding both epigastric pain at 48 hours (t=2.908, p=.005), 72 hours (t=3.010, p=.004), but there was a significant difference in objective body temperature discomfort (t=2.895, p=.008). CONCLUSION: Thermotherapy relieved shoulder gas-related pain and objective body temperature discomfort. It needs to be developed and applied to improve post-operative discomfort in patients with laparoscopic hysterectomies.

A metabolomics study was conducted to identify urinary biomarkers for breast cancer, using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS), analyzed by principal components analy-sis (PCA) as well as a partial least squares-discriminant analysis (PLS-DA) for a metabolic pattern analysis. To find potentialbiomarkers, urine samples were collected from before- and after-mastectomy of breast cancer patients and healthy controls.Androgens, corticoids, estrogens, nucleosides, and polyols were quantitatively measured and urinary metabolic profiles wereconstructed through PCA and PLS-DA. The possible biomarkers were discriminated from quantified targeted metabolites with ametabolic pattern analysis and subsequent screening. We identified two biomarkers for breast cancer in urine, β-cortol and 5-methyl-2-deoxycytidine, which were categorized at significant levels in a student t-test (p-value < 0.05). The concentrations ofthese metabolites in breast cancer patients significantly increased relative to those of controls and patients after mastectomy.Biomarkers identified in this study were highly related to metabolites causing oxidative DNA damage in the endogenous metab-olism. These biomarkers are not only useful for diagnostics and patient stratification but can be mapped on a biochemical chartto identify the corresponding enzyme for target identification via metabolomics.Key words: Metabolomics, Breast Cancer, Biomarker, PLS-DA, Urine

In this study, total mercury and methyl mercury in whole blood of Korean was analyzed so as to investigate the correlation between total mercury (T-Hg) and methyl mercury (Me-Hg). 4000 whole blood samples were divided in four groups, according to T-Hg concentration in percentile: group I (p25-p50), group II (p50-p75), group III (p75-p95) and group IV (p95-p100). 100 samples were randomly selected from the each group, and Me-Hg concentration was measured. T-Hg concentration in whole blood was analyzed using a Direct Mercury Analyzer-80 and obtained limit of detection (LOD) was 0.2 µgL �1 . Me-Hg concentration was analyzed with ethylate derivatization using headspace-gas chromatography-mass spectrometry, and obtained LOD of methyl mercury was 0.5 µgL �1 . The geometric means of T-Hg and Me-Hg were 6.35 µgL �1 and 4.44 µgL �1 , respectively, and 71.91% of T-Hg was presented as Me-Hg.

Di(2-ethylhexyl) phthalate (DEHP) is one of the common phthalate plasticizers used primarily in soft polyvinyl chloride, which is a plastic polymer that makes up the total weight of goods from 1% up to 40% in many consumer products. The aims of this study were to examine the urinary DEHP metabolites in South Korean children and to investigate the correlation between mother and child DEHP urine excretion. Three kinds of urinary DEHP metabolites: mono(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (5-OH-MEHP) and mono(2-ethyl-5-oxohexyl) phthalate (5-oxo-MEHP), were analyzed. The total of 954 samples (nChildren=392, nMothers=265, nAadults=297), including 258 mother and child pairs, were analyzed using isotope dilution gas chromatography–mass spectrometry. Many studies present higher concentration of DEHP metabolites detected from adults in reproductive age than adults in other ages. Therefore, adults who are age-matched to mothers were evaluated to serve as a standard of comparison against mothers. All statistical analysis was made by adjusting detected volume concentrations (μg/L) with respect to creatinine concentrations (mg/dL) since urinary DEHP metabolites were studied using human reference. The difference in median levels of sum of urinary DEHP metabolites was only significant when children were analyzed in relation to region (p-value≤0.005). Among the three DEHP metabolites, only MEHP of children was significantly correlated to that of paired mothers (p-value≤0.01). The present paper defines the relative metabolic rate (RMR) of DEHP metabolism for the first time in study on phthalates. Children had faster RMR than mothers and adults, specifically in the first step of DEHP metabolism (RMR1: MEHP hydroxylation to 5-OH-MEHP), and RMR1 of children between 1 and 24 months was the fastest. The above results may be used to study and assess human health risk from DEHP exposures, especially among mothers and children in Korea. Keywords: DEHP metabolites, Human exposure, Children, Biomonitoring

The study aim was to evaluate the correlation of sexual dimorphism and pubertal stage with steroid metabolism in adolescents.The serum levels of cholesterol, pregnenolone, and dehydroepiandrosterone (DHEA) and their sulfate conjugates were quantitatively profiled from serum samples of 199 adolescents (87 boys and 112 girls), aged 6 to 14years.In the prepubertal stage, DHEA levels in girls were higher than those in boys; however, significantly increased DHEA levels during pubertal development in boys. Pregnenolone levels were significantly higher in girls; however, the levels of its sulfate were higher in boys. The serum levels of both cholesterol and its sulfate were higher in boys, especially in the early to mid-pubertal stages. DHEA and DHEA sulfate levels in both sexes significantly increased with pubertal development (P for trend0.05), while pregnenolone, cholesterol, and cholesterol sulfate in both sexes were stable. The metabolic ratios, indicating sulfotransferase activity, were significantly higher in boys, and increased with pubertal development in boys, but not in girls, while CYP11A1 activity levels increased significantly in both sexes with pubertal development.Sexual dimorphism in key enzymes of androgen biosynthesis during pubertal changes may help elucidate the normal physiology of steroidogenesis.

Department of Environmental Health, Korea University, Seoul 136-701, KoreaReceived October 15, 2011, Accepted September 4, 2012Headspace (HS) and headspace solid-phase microextraction (HS-SPME) were studied for extracting volatileorganic compounds (VOCs) from whole blood, with chemical and instrumental variables being optimized formaximum sensitivity: incubation at 60 °C, equilibration for 30 min, pH 11, and 2 mL injection volume. Bothtechniques provided accurate analyses, with detection limits of 0.05-0.1 ng mL

Fat malabsorption is an important cause of poor growth in infancy and childhood. Steatorrhea tests have been devel- oped using various methods. Traditional measurements of stool fat, however, require large samples and it often takes as a week to complete the analysis. In this paper, a liquid chromatography-electrospray ionization/mass spectrometry (LC-ESI/MS) method was developed for simultaneous quantitative analysis of triacylglycerols, triolein, diolein and monoolein, in mouse feces. More- over, the procedure was rapid, simple as well as compatible with LC-ESI/MS. Chloroform-isopropyl alcohol solution was used for fat-soluble sample extraction. After centrifugation and filtration, an analytical solution was prepared. Triolein, diolein and monoolein were separated using non-aqueous reversed-phase column with the mobile phase consisting of A (methanol) and B (acetone-isopropyl alcohol). The precision (% CV) and accuracy (% bias) of the assay were 3.8-14.7% and 85.2-114.9%, respec- tively. This method has been successfully applied to simultaneous determination of triolein, diolein and monoolein in feces from 30 mice. This method can therefore be applied to measure triacylglycerols in mouse feces accurately and precisely by LC-ESI/ MS, thereby helping to predictive biomarker in fat malabsorption and diagnostic research.

Future Convergence Research Division, Korea Institute of Science and Technology, Seoul 136-791, KoreabDepartment of Biomolecular Science, University of Science and Technology, Daejeon 305-333, KoreaReceived March 3, 2012; Revised March 7, 2012; Accepted March 7, 2012First published on the web March 15, 2012; DOI: 10.5478/MSL.2012.3.1.004

Many clinical events result in specific and characteristic alterations that originate in biochemical imbalances and are measurable in biological fluids and tissues. Metabolomics consists of the qualitative and quantitative analysis of intraand intercellular metabolites using 2 different analytical methods: (1) non-targeted metabolite profiling and (2)targeted metabolite profiling. Non-targeted metabolite profiling involves the identification and characterization of a large number of metabolites and their precursors. In contrast,targeted metabolite profiling focuses on quantitative changes in metabolites of interest (e.g., amino acids, carbohydrates, steroids, and fatty acids) based on a priori knowledge of the biological function or metabolic pathway. Steroid hormones regulate human body functions and are components of the endocrine system, which coordinates physiological and behavioral responses for specific biological purposes, such as reproduction. Steroids are active at very low concentrations and have a range of biological effects (e.g., homeostasis). Steroid analysis has gained importance in biomedical, pharmaceutical, engineering, and agricultural sciences. Clinical assays for steroid analysis must be robust, highly sensitive, automated, and simple. Steroids analysis is used in the clinical events for the diagnosis of stress,1 Cushing’s syndrome,2 hirsutism,3congenital steroid enzyme deficiency,4 hypertension,5 and as a doping test.6 Biological specimens used in clinical applications include urine, serum, and saliva, and they exhibit some behaviors of steroids (e.g., free forms, conjugated forms, protein-binding forms). Steroids in biological specimens behave differently and different considerations are required when developing methods for their analysis.7 Due to its high sensitivity and selectivity, gas chromatographymass spectrometry (GC-MS) is used to analyze metabolites and precursors of steroid hormones. In recent, GC-MSbased steroid phenol-typing has demonstrated diagnostic power and the potential to discover metabolic diseases.8 This review focuses on the methods involved in a metabolomic study based on GC-MS analysis of steroid hormones in biological specimens. Sample preparation procedures and their major steps are discussed in detail.The various techniques and materials that have been used to achieve efficient analytical results during the past decade are described. Comparative studies of crucial aspects are included to facilitate the understanding of the developed methods.

5α-Dihydrotestosterone (DHT) is the primary active metabolite of testosterone, catalyzed by 5α-reductase (5αR) in the skin, prostate, and liver. In this study, the 5αR activity in rat liver S9 fraction in the presence of a NADPH-generating system was evaluated and compared by gas chromatography-mass spectrometry (GC-MS)-based in vitro assays. Testosterone and a 5αR inhibitor, finasteride, were added to the S9 fractions and incubated at 37oC for 1 h. Both testosterone and DHT were quantitatively measured and compared with two different GC-MS-based steroid profiling techniques. DHT was not detected by conventional GC-MS analysis in the absence of finasteride when the concentration of testosterone in the S9 fraction was less than 0.2 μM, whereas the isotope-dilution GC-MS (GC-IDMS) system was able to evaluate the 5αR activity. Because the S9 fraction contains more reactive enzymes and is easier to collect from tissues compared with a microsomal solution, the combination of the S9 fraction and GC-IDMS technique may be a promising assay for evaluating the 5αR activity in large-scale clinical studies.

Pyrophosphates are the key intermediates in the biosynthesis of isoprenoids, and their concentrations could reveal the benefits of statins in cardiovascular diseases. Quantitative analysis of five pyrophosphates, including isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate (DMAPP), geranyl pyrophosphate (GPP), farnesyl pyrophosphate (FPP), and geranylgeranyl pyrophosphate (GGPP), was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in negative ion- ization mode. After dilution with methanol, samples were separated on a 3 µm particle multi-modal C18 column (50 ×2m m) and quantified within 10 min. The gradient elution consists of 10 mM ammonium bicarbonate and 0.5% triethylamine (TEA) in water and 0.1% TEA in 80% acetonitrile was used at the flow rate of 0.4 mL/min. Overall recoveries were 51.4-106.6%, while the limit of quantification was 0.05 µg/mL for GPP and FPP and 0.1 µg/mL for IPP, DMAPP, and GGPP. The precision (% CV) and accuracy (% bias) of the assay were 1.9-12.3% and 89.6-111.8%, respectively, in 0.05-10 µg/mL calibration ranges (R 2 > 0.993). The devised LC-MS/MS technique with the multi-modal C18 column can be used to estimate the biological activity of pyrophosphates in plasma and may be applicable to cardiovascular events with cholesterol metabolism as well as the drug efficacy of statins.

Pyrophosphates are the key intermediates in the biosynthesis of isoprenoids, and their concentrations could revealthe benefits of statins in cardiovascular diseases. Quantitative analysis of five pyrophosphates, including isopentenyl pyrophosphate(IPP), dimethylallyl pyrophosphate (DMAPP), geranyl pyrophosphate (GPP), farnesyl pyrophosphate (FPP), and geranylgeranylpyrophosphate (GGPP), was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in negative ionizationmode. After dilution with methanol, samples were separated on a 3 μm particle multi-modal C18 column (50 × 2 mm)and quantified within 10 min. The gradient elution consists of 10 mM ammonium bicarbonate and 0.5% triethylamine (TEA) inwater and 0.1% TEA in 80% acetonitrile was used at the flow rate of 0.4 mL/min. Overall recoveries were 51.4-106.6%, whilethe limit of quantification was 0.05 μg/mL for GPP and FPP and 0.1 μg/mL for IPP, DMAPP, and GGPP. The precision (% CV)and accuracy (% bias) of the assay were 1.9-12.3% and 89.6-111.8%, respectively, in 0.05-10 μg/mL calibration ranges (R2 > 0.993).The devised LC-MS/MS technique with the multi-modal C18 column can be used to estimate the biological activity of pyrophosphatesin plasma and may be applicable to cardiovascular events with cholesterol metabolism as well as the drug efficacy of statins.

A distributed Bragg reflector (DBR) utilizing the digital alloy Al0.9Ga0.1As was grown by using the molecular beam epitaxy (MBE) method for application in 1.3 μm optical communications and compared to the analog-alloy AlGaAs/GaAs DBR. The transmission electron microscopic (TEM) image showed a highly abrupt boundary of AlAs and GaAs, which supports the formation of a digital-alloy Al0.9Ga0.1As layer. The measurement showed that the digital-alloy AlGaAs/GaAs DBR had similar reflection spectra with enhanced uniform distribution over the whole substrate surface compared to the analog-alloy one. In the digital-alloy Al0.9Ga0.1As/GaAs DBR cavity, the reflection dip position was measured at around 1273 nm and the standard deviation of the distribution of the reflection dip was 1.31 nm in wavelength over 1/4 of a three-inch wafer.

Metabolomics has focused on toxicological applications to (1) understand the mechanisms of toxicity, (2) identify novel biomarkers of toxicity, and (3) provide in vivo assessment in animal models through simple and fast methods to date. The toxicological effects of nonylphenol (NP) were evaluated after intraperitoneal injection of rats with 0, 50, and 250 mg kg(-1) day(-1) NP for four consecutive days. In the nontargeted approach, different extraction conditions were introduced to investigate the effects of NP on rats through gas chromatography/mass spectrometry (GC/MS). The GC/MS data obtained were further analyzed with partial least-squares discriminant analysis to compare toxicological effects between control and treated groups. The targeted approach was also used in combination with GC/MS to quantify endocrine hormones and to identify possible biomarkers in rat urine under optimal extraction conditions. In addition, we considered the metabolic trajectory to examine the metabolite profiles and patterns related to steroid metabolism in rats that were treated with NP, considering both treatment amount and time. The data suggest that tetrahydrocorticosterone and 5alpha-tetrahydrocorticosterone are possible urinary biomarkers of NP-induced toxicity. This metabolomic approach is a promising tool to assist with screening in toxicological studies.

We report transport measurements through an in-plane-gate quantum dot transistor (IPGQDT). Our IPGQDT has etched trench isolation between the two-dimensional electron gas QD and IPGs, and it operates in enhancement mode. At relatively small IPG biases, we observe negative differential resistance (NDR) in the current–voltage characteristics. The position of the NDR peak is controlled systematically by the change of the IPG bias. At large biases, the IPGQDT exhibits single-electron tunneling. All of these transport data are not sample specific and consistent with the size of the QD.

We investigated a photovoltaic three-stacked In"0"."5Ga"0"."5As/GaAs quantum dot infrared detector (QDIP) with an Al"0"."3Ga"0"."7As single-sided blocking layer. We observed broad photocurrent spectra in the photon energy range of 120-400 meV (@l~3-10@mm) at zero-bias voltage, due to the photovoltaic effect at low temperatures. The peak responsivity was about 10.5mA/W at a photon energy of 200 meV (@l~6.2@mm) at T=40K. The large photovoltaic effect in our detector was a result of the enhanced asymmetric band structure, caused not only by the segregation of highly doped Si atoms, but also by the single-sided Al"0"."3Ga"0"."7As layer beneath the top contact layer.

Optical and electrical characteristics of n–i–n InGaAs/GaAs quantum-dot (QD) infrared photodetectors are reported. In particular, the low-frequency excess electrical noise is measured at room temperature and analyzed in conjunction with the optical properties of the structure. The three stackings of QD were formed by atomic layer molecular-beam epitaxy and highly Si-doped, and AlGaAs current-blocking layer was also included to reduce the dark current. The power-dependent photoluminescence (PL) spectra at 300 K indicates that there are at least three confined states in the QD. The photo-current was observed only at low temperatures (10 K) at wavelengths between 3 and 9 μm with three peaks. The dark current was relatively large and asymmetric at low temperatures. At room temperature the dark current was symmetric and ohmic. The 1/ f -like low-frequency noise spectral density exhibited an almost quadratic current dependence giving a large value of the Hooge parameter of the order of unity. The relatively low-growth temperature for the AlGaAs current blocking layer and the high doping at the quantum dots seem to generate a considerable amount of defects and result in low-temperature photodetection and a large low-frequency noise density.

Interest in the analysis of low abundance neuropeptides particularly using matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI–TOF–MS) is increasing because these neuropeptides are essential to the mechanism of transportation and the metabolism. This article describes an immunoprecipitation procedure that is suitable for MALDI–MS analysis of substance P (SP), a neuropeptide, in rat brain tissues. Substance P was precipitated from brain tissue extracts by immunoprecipitation with antibodies directed against SP, and are analyzed by MALDI–TOF–MS. Mass spectrometric analysis showed a singly charged [M+H]+ ion peak that corresponded to the SP molecular mass and was observed with a detection error of 1.6%. The average mass errors between the observed and theoretical molecular mass were within the 0.11 Da range. Capillary zone electrophoresis analysis was subsequently performed, and the effects of the different separation parameters were examined. Beginning with milligram quantities of brain tissue, picomole quantities of SP could be detected using this method.

We recently reported that treatment with a pharmacologic dose of recombinant human growth hormone (GH) resulted in a significant loss of body fat and gain in lean tissue in HIV-infected patients with syndromes of fat accumulation. However, insulin-mediated glucose disposal decreased transiently after one month of GH therapy. The present paper focuses on the changes of hepatic carbohydrate and fat metabolism associated with GH treatment in the same subjects. We assessed hepatic insulin sensitivity under both fasting and hyperinsulinemic-euglycemic clamp conditions prior to and after one and six months of GH treatment (3 mg/day) in five patients using stable isotope tracer techniques. Indirect calorimetry, and measurements of lipid concentrations. Fasting endogenous glucose production (EGP) increased significantly at one month (12.0 ± 0.7 to 14.9 ± 0.9 μmol/kg/min, P < 0.03), and the increase was sustained at six months of GH treatment (14.0 ± 1.1μ mol/kg/min, NS). This increase in EGP was driven in part by increased glucogenesis (GNG) (3.5 ± 0.9 to 5.2 ± 0.9 and 5.8 ±1.2 μmol/kg/min, n = 4, P < 0.01 and P < 0.01 at one and six months, respectively); small changes in hepatic glycogenolysis also contributed. Sustained increases in lipolysis and progressive decreases in hepatic fractional de novo lipogenesis (DNL) and triglyceride concentrations occurred with GH treatment. These changes were accompanied by an improved lipid profile with a significant increase in HDL cholesterol and significant decreases in total and LDL cholesterol and triglyceride levels, the latter consistent with the decrease in hepatic DNL. During a hyperinsulinemic-euglycemic glucose clamp, EGP and GNG were markedly suppressed compared to the corresponding time points under fasting conditions, albeit less so when measured after one month of GH treatment. Thus, in HIV-infected patients with abnormal fat distribution, pharmacologic doses of GH improved the overall lipid profile, but worsened glucose homeostasis under both fasting and hyperinsulinemic conditions. The combined implications of these positive and negative metabolic effects for cardiovascular disease risk remain unknown.