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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.

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.

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.

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.

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

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.

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.

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 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.

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 naive 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 naive 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.

Drug-induced cytochrome P450 (CYP) activity affects endocrine function and drug clearance rates, leading to the development of unpredictable pathologic and toxicologic risks.Urinary steroid profiling based on gas chromatography-mass spectrometry (GC-MS) was used for simultaneous quantification of CYP-mediated regioselective hydroxysteroids and their substrates, including 26 androgens, 9 estrogens, 5 progestins, and 7 corticoids. The quantitative data were visualized using a hierarchically clustered heat map to allow identification of CYP-mediated steroid signatures. Twelve healthy subjects were orally administered 600 mg of rifampicin a day for 7 days, and their CYP enzyme activity was evaluated.Using GC-MS, all 47 steroids were well separated with good peak shapes. This assay had good linearity (r0.994) in a dynamic range, and the interassay imprecision (% CV) and inaccuracy (% bias) were 3.0%-15.6% and 98.0%-109.2%, respectively. Administration of the CYP3A4 inducer rifampicin produced distinct differences in CYP3A4 and CYP11B1, CYP19A1, HSD11B, and HSD17B, which were indicated by their heat map-visualized steroid signatures.This CYP-mediated steroid signature profile allows simultaneous assessment of CYP1A, CYP1B, CYP2C, CYP3A, CYP11B, CYP17A, CYP19A, and CYP21A in urine samples. This method could therefore be a useful tool for assessing drug efficacy.

The short- and long-term effects of a single exposure to gamma radiation on steroid metabolism were investigated in mice. Gas chromatography–mass spectrometry was used to generate quantitative profiles of serum steroid levels in mice that had undergone total-body irradiation (TBI) at doses of 0 Gy, 1 Gy, and 4 Gy. Following TBI, serum samples were collected at the pre-dose time point and 1, 3, 6, and 9 months after TBI. Serum levels of progestins, progesterone, 5β-DHP, 5α-DHP, and 20α-DHP showed a significant down-regulation following short-term exposure to 4 Gy, with the exception of 20α-DHP, which was significantly decreased at each of the time points measured. The corticosteroids 5α-THDOC and 5α-DHB were significantly elevated at each of the time points measured after exposure to either 1 or 4 Gy. Among the sterols, 24S-OH-cholestoerol showed a dose-related elevation after irradiation that reached significance in the high dose group at the 6- and 9-month time points.

Bisphenol-A (BPA) possesses estrogenic properties both in vitro and in vivo as an endocrine disrupting chemical. Humans experience a long-term and cumulative exposure to BPA. BPA was detectable in 97.3% of 1904 urine specimens from Korean adults. We investigated urinary estrogen concentrations in subjects with low and high BPA concentrations and its possible association with estrogen metabolism. Urine samples were collected from a high BPA concentration group (BPA-H; n=100, 11.05 ± 20.47 μg/g creatinine) and a low BPA concentration group (BPA-L; n=100, 0.70 ± 0.22 μg/g creatinine) from Korea Biomonitoring Program of Hazardous Materials Survey 2009-2010. Urinary estrogens were enzymatically hydrolyzed, extracted, and then derivatized for quantitative analysis by gas chromatography-mass spectrometry. Estrogen levels were higher in the BPA-H group than in the BPA-L group. Concentrations of estrone, 17β-estradiol, and their hydroxylated metabolites in both men and women were significantly higher in the BPA-H group than in the BPA-L group (p0.04). Furthermore, in the BPA-H group, estrogen metabolism to 4-hydroxy-estrone and 4-hydroxy-17β-estradiol was more active than that to 2-hydroxy-estrone and 2-hydroxy-17β-estradiol. Although single measurement and/or single spot urine samples limit the measurement of long-term exposure to BPA, we found significant differences of estrogen metabolism in the BPA-H and the BPA-L groups. The increase of hydroxyestrogens, especially 4-hydroxyestrogens, can be an important factor resulting negative effects of prolonged exposure to BPA.

Precocious puberty (PP) refers to the appearance of physical and hormonal signs of pubertal development at an abnormally early age. Urinary steroid signatures obtained from 42 patients with central PP and 40 patients with peripheral PP were assessed to compare metabolic changes. Levels of androgens such as testosterone, androstenedione, androstenediol, 16α-hydroxy-dehydroepiandrosterone, and 5α-androstenedione tended to be high in both PP groups, and the level of 17β-estradiol was higher in the central-PP group (P

On December 7th, 2007, the Hong Kong tanker Hebei Spirit (HS) (146,848 tons) was crushed by a crane ship near the shore of Taean, Korea. More than 12,547 kl of crude oil spilled into the sea and contaminated the western coastline of the Korean peninsula. For a period of six months after the accident, approximately 1,000,000 volunteers participated in the cleanup. Our goal in this study was to examine the exposure status and acute health effects on volunteers that participated in the oil spill cleanup.A survey questionnaire was filled out by 565 volunteers, requesting information regarding physical symptoms. Out of the total number of participants, urine samples from 105 university student volunteers were collected before and after the cleanup work, and metabolite levels of volatile organic compounds and polycyclic aromatic hydrocarbons were analyzed.Volunteers that participated for longer cleanup work reported an increase in physical symptoms including visual disturbance, nasal and bronchus irritation, headaches, heart palpitations, fatigue and fever, memory and cognitive disturbance, and abdominal pain. The levels of t,t-muconic acid, mandelic acid, and 1-hydroxypyrene were significantly higher in samples after cleanup than those measured before participation (p0.05). Other than the associated risk of dermal irritation with the difference in the t,t-muconic acid level between the post- to pre-cleanup levels, no other physical symptoms demonstrated a significant association with changes observed in the levels of urinary metabolites.Based on the significant increase of subjective symptoms in volunteers participating in the study, monitoring of the long term health effects, focusing on those with longer exposure, is warranted.

Capillary electrophoresis–electrospray tandem mass spectrometry (CE-ESI/MS/MS) is a simple and highly sensitive method for quantifying seven urinary androgen glucuronides. The urine samples were diluted and filtered through a membrane filter, and the filtrate was injected into a CE-MS/MS system without further sample preparation steps such as extraction and derivatization. The calibration ranges were 0.01–5 µg/mL for glucuronides of androsterone and 11β-OHAn-3G, and 5–500 ng/mL for glucuronides of 11-ketoAn, DHEA, testosterone, epitestosterone and DHT. The linearity of the method was 0.992–0.998, and the limits-of-detection at a signal-to-noise ratio of 3 were 5–10 ng/mL. The coefficients of variation were in the range of 4.0–9.0% for intra-day assay and 4.1–9.8% for inter-day assay. The proposed method may be applicable to metabolic profiling in both quantitative and qualitative analysis. Copyright © 2008 John Wiley & Sons, Ltd.

The metabolomic approaches for mining biomarkers of women's cancers based on gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry combined with partial least squares-discriminant analysis are described.To identify urinary potential biomarkers, the qualitative and quantitative analyses were introduced with 10 breast, 9 ovarian and 12 cervical cancer patients as well as 22 normal controls, which were considered with their ages and menopausal state.For comprehensive metabolomic approaches, the non-targeted qualitative profiling was first achieved to get metabolic patterns of collected samples and the targeted quantitative analysis focused on hormonal metabolism was also conducted. Two known biomarkers, i.e., 5-hydroxymethyl-2-deoxyuridine and 8-hydroxy-2-deoxyguanosine, in breast cancer were also confirmed using the present methods. In addition, 3 potential biomarkers for ovarian cancer i.e. 1-methyladenosine, 3-methyluridine, and 4-androstene-3,17-dione, which were categorized in significantly increased level using one way of variance analysis (p0.05), were identified as quantitatively targeted metabolites with pattern analysis. The cancer markers identified in this study are highly related to metabolites which are responsible for oxidative DNA damage and DNA methylation process.The present metabolomic approaches are not only useful for diagnostic tools and patient stratification, but may be mapped on metabolic network to reflect disease states.

In the present study, we simultaneously measured several polyols, such as adonitol, arabitol, dulcitol, glucose, myo-inositol, mannitol, sorbitol, and xylitol, in urine by gas chromatography/mass spectrometry–positive chemical ionization. We also examined possible relationship between the levels of these metabolites and age in normal individuals. In order to proceed to its quantification by GC/MS, 200 μL of a urine sample were diluted with 3 mL of distilled water, lyophilized, acetylated, and then analyzed them. Using this method, we were able to quantify as little as 0.5–1.0 ng/μL, and we made the calibration curves to be linear from 0.25 to 250 ng/μL (r2 > 0.991). Analytical recoveries were over 89.4%, and the inter-day and intra-day variability for accuracy and reproducibility was less than 20%. In the normal urine sample, the levels of polyols were gender-differentiated and age-related. This simple GC/MS method is sensitive and allows the measurement of wide ranges of polyols using small amounts of urine. We conclude that the quantitation of urinary polyols using GC/MS appears to be a clinically useful method for assessing polyol-pathway activity.

In the absence of HIV infection, changes in adipose tissue and lipid levels, HIV protease inhibitor therapy increases fasting glucose levels, suggestive of hepatic insulin resistance. After 4 weeks of indinavir treatment in nine HIV-negative healthy men, fasting glucose production and glycogenolysis were significantly increased. During the euglycemic hyperinsulinemic clamp, indinavir blunted the ability of insulin to suppress glucose production. Therefore, indinavir worsens hepatic insulin sensitivity, increasing endogenous glucose production.

Objectives To assess the protective effects of wearing protective devices among the residents and volunteers who participated in the cleanup of the Hebei Spirit oil spill. Methods A total of 288 residents and 724 volunteers were surveyed about symptoms, whether they were wearing protective devices and potential confounding variables. The questionnaires were administered from the second to the sixth week following the accident. Spot urine samples were collected and analyzed for metabolites of 4 volatile organic compounds (VOCs), 2 polycyclic aromatic hydrocarbons (PAHs), and 6 heavy metals. The association between the wearing of protective devices and various symptoms was assessed using a multiple logistic regression adjusted for confounding variables. A multiple generalized linear regression model adjusted for the covariates was used to test for a difference in least-square mean concentration of urinary biomarkers between residents who wore protective devices and those who did not. Results Thirty nine to 98% of the residents and 62-98% of volunteers wore protective devices. Levels of fatigue and fever were higher among residents not wearing masks than among those who did wear masks (odds ratio 4.5; 95% confidence interval 1.23-19.86). Urinary mercury levels were found to be significantly higher among residents not wearing work clothes or boots (p Conclusions Because the survey was not performed during the initial high-exposure period, no significant difference was found in metabolite levels between people who wore protective devices and those who did not, except for mercury, whose biological half-life is more than 6 weeks.