Metabolomic Analysis Identifies Alterations of Amino Acid Metabolome Signatures in the Postmortem Brain of Alzheimer's Disease

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.

Liquid Chromatography-Mass Spectrometry-Based In Vitro Metabolic Profiling Reveals Altered Enzyme Expressions in Eicosanoid Metabolism

BACKGROUND: Eicosanoids are metabolites of arachidonic acid that are rapidly biosynthesized and degraded during inflammation, and their metabolic changes reveal altered enzyme expression following drug treatment. We developed an eicosanoid profiling method and evaluated their changes on drug treatment. METHODS: Simultaneous quantitative profiling of 32 eicosanoids in liver S9 fractions obtained from rabbits with carrageenan-induced inflammation was performed and validated by liquid chromatography-mass spectrometry coupled to anion-exchange solid-phase purification. RESULTS: The limit of quantification for the devised method ranged from 0.5 to 20.0 ng/mg protein, and calibration linearity was achieved (R 2>0.99). The precision (% CV) and accuracy (% bias) ranged from 4.7 to 10.3% and 88.4 to 110.9%, respectively, and overall recoveries ranged from 58.0 to 105.3%. Our method was then applied and showed that epitestosterone treatment reduced the levels of all eicosanoids that were generated by cyclooxygenases and lipoxygenases. CONCLUSIONS: Quantitative eicosanoid profiling combined with in vitro metabolic assays may be useful for evaluating metabolic changes affected by drugs during eicosanoid metabolism.

Up-regulation of BLT2 is critical for the survival of bladder cancer cells

The incidence rates of urinary bladder cancer continue to rise yearly, and thus new therapeutic approaches and early diagnostic markers for bladder cancer are urgently needed. Thus, identifying the key mediators and molecular mechanisms responsible for the survival of bladder cancer has valuable implications for the development of therapy. In this study, the role of BLT2, a receptor for leukotriene B4 (LTB4) and 12(S)-hydroxyeicosatetraenoic acid (HETE), in the survival of bladder cancer 253J-BV cells was investigated. We found that the expression of BLT2 is highly elevated in bladder cancer cells. Also, we observed that blockade of BLT2 with an antagonist or BLT2 siRNA resulted in cell cycle arrest and apoptotic cell death, suggesting a role of BLT2 in the survival of human bladder cancer 253J-BV cells. Further experiments aimed at elucidating the mechanism by which BLT2 mediates survival revealed that enhanced level of reactive oxygen species (ROS) are generated via a BLT2-dependent up-regulation of NADPH oxidase members NOX1 and NOX4. Additionally, we observed that inhibition of ROS generation by either NOX1/4 siRNAs or treatment with an ROS-scavenging agent results in apoptotic cell death in 253J-BV bladder cancer cells. These results demonstrated that a 'BLT2-NOX1/4-ROS' cascade plays a role in the survival of this aggressive bladder cancer cells, thus pointing to BLT2 as a potential target for anti-bladder cancer therapy.

The Effects of Wearing Protective Devices among Residents and Volunteers Participating in the Cleanup of the Hebei Spirit Oil Spill / 예방의학회지

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<0.05). 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.

Steroid Hormone Metabolism in Patients with Pelvic Organ Prolapse / 대한산부인과학회잡지

OBJECTIVE: To identify 1) whether the endogenous steroid hormone metabolism in patients with pelvic organ prolapse was different from that of normal women, 2) the relationship between endogenous steroid hormone metabolites and the stage of the pelvic organ prolapse. METHODS: Twenty postmenopausal women who were clinically diagnosed as having pelvic organ prolapse and 20 volunteer postmenopausal women not having pelvic organ prolapse were included in the study. We compared the urinary profiles of endogenous steroids between the two groups and investigated the relationship between urinary profiles of the endogenous steroids and the degree of pelvic organ prolapse. Urinary profiles of the endogenous steroids were assayed by gas chromatography-mass spectrometry. RESULTS: The ages of the patients and control group were 64.6 +/- 6.5 and 63.5 +/- 3.9 years, and the Body Mass Index (BMI) was 23.96 +/- 3.14 and 24.11 +/- 2.73 kg/m2 in patients and in normal subjects, respectively. The number of patients in each stage were 4 in stage I, 4 in stage II, 6 in stage III and 6 in stage IV. 5-androstene-3beta, 16beta, 17beta-triol (5-AT), 11beta-hydroxy androstenedione (An) and 17beta-estradiol were significantly increased in patients with pelvic organ prolapse over that of the control group (0.76 +/- 0.67 vs 0.06 +/- 0.03 micro mole/g creatinine; p=0.002, 1.16 +/- 0.83 vs 0.65 +/- 0.23 micro mole/g creatinine; p=0.04, 15.08 +/- 9.81 vs 8.53 +/- 6.19 micro mole/g creatinine; p=0.04). However, tetrahydrocortisone (THE) was significantly increased in the control group over that in patients having pelvic organ prolapse (9.80 +/- 6.21 vs 5.22 +/- 4.89 micro mole/g creatinine; p=0.04). The androgen metabolites, 5-AT and THE significantly correlated with the POP-Q stage (R=0.418; p=0.027, R=0.46; p=0.016). Among the estrogen metabolites, 17beta-estradiol was correlated to the POP-Q stage but not mathematically significantly (R=0.38; p=0.05) and the 17beta-estradiol/estrone ratio weakly correlated to pelvic organ prolapse stage (R=0.14; p=0.49), by showing a low correlation coefficiency. CONCLUSION: The urinary concentrations of 17beta-estradiol, 5-AT and 11beta-hydroxy An increased in patients with pelvic organ prolapse over that of the control group and 5-AT, THE and 17beta-estradiol showed a relationship to the progression of pelvic organ prolapse in Korean women. The metabolites of endogenous steroid hormones could be contributing factors in the pathogenesis of pelvic organ prolapse.

Relationship between Urinary Endogenous Steroid Metabolites and Lower Urinary Tract Function in Postmenopausal Women

To investigate the relationship between the endogenous steroid hormones and the lower urinary tract function in postmenopausal women. Thirty postmeopausal volunteer women who did not have lower urinary tract symptoms or hormone replacement therapy were enrolled in this study. Urodynamic studies included uroflowmetry, multi-channel cystometry, and urethral pressure profilometry were conducted. Gas Chromatography- Mass Spectroscopy (GC-MS) was used to measure the urinary endogenous steroid hormone metabolites. The relationship between the urinary profile of the endogenous steroids and the urodynamic parameters of these patients were investigated. The mean ages of the patients were 60.6 +/- 5.5 years, and the Body Mass Index (BMI) averaged 24.56 +/- 2.23 (kg/m2). Of the progesterone metabolites, pregnandiol was significantly related to the residual volume in the uroflowmetry and the functional urethral length parameters (R=0.98, p=0.000; R= -0.65, p=0.04). Pregnantriol was significantly related to the maximum flow rate, the residual volume in uroflowmetry, the maximum urethral closure pressure and the functional urethral length (R=-0.64, p=0.04; R=0.82, p=0.01; R=0.04, p=0.04; R=- 0.79, p=0.01). In the androgen metabolites, androstenedione, 5-AT, 11- keto Et, 11-betahydroxy Et, THS, and THE were significantly related to the residual volume in uroflowmetry (R=0.92, p=0.001; R=0.84, p=0.008; R=0.99, p=0.000; R=0.72, p=0.03; R=0.97, p=0.000; R=0.85, p=0.00). beta-THF/alpha-THF was significantly related to the maximum flow rate, the residual volume in uroflowmetry, the maximum urethral closure pressure and the functional urethral length (R=-0.76, p=0.02; R=0.67, p=0.04; R=0.74, p=0.02; R=-0.92, p=0.000). alpha-cortol was significantly related to the residual volume in uroflowmetry, the maximum urethral closure pressure and the functional urethral length (R=0.81, p=0.01; R=0.71, p=0.03; R=-0.87, p=0.000). Of the estrogen metabolites, estrone (E1) was significantly related to the normal desire to void (R=0.68, p=0.04) and 17 beta-estradiol/estrone was also significantly related to the normal and strong desire to void (R=-0.70, p=0.03 and R=-0.74, p=0.02, respectively). The urinary progesterone and androgen metabolite concentrations were positively related to the residual volume in uroflowmetry and positively or negatively related to MUCP and FUL. However, the urinary estrone concentration was positively related to the normal desire to void and 17 beta-estradiol/estrone was significantly related to the normal and strong desire to void.

Expression of Neuroendocrine Cells in Benign Prostatic Hyperplasia and the Effect of Dihydrotestosterone / 대한비뇨기과학회지

PURPOSE: Neuroendocrine (NE) cells of the prostate are considered to be involved in the pathogenesis of benign prostate hyperplasia (BPH). By a comparative analysis of NE cell density in BPH tissue of men who were either exposed to or not exposed to 5alpha-reductase inhibitor, we investigated the relationship between NE cells and BPH, and the effect of androgen deprivation on NE cells. MATERIALS AND METHODS: Prostate tissue specimens, obtained from 30 men by transurethral resection of the prostate or radical cystoprostatectomy, were used. Of the 30 patients, 10 had a prostate smaller than 25 ml (normal control), the other 20 had a prostate larger than 40ml, 10 of who had taken 5alpha-reductase inhibitor (finasteride) for 3 months before surgery (androgen blockade group), and 10 who had not (BPH group). The distribution of NE cells in the prostate was examined using the anti-chromogranin A (CgA) antibody, and the density of the CgA-positive cells was compared by an optical dissector method. Immunoblotting was performed using the neuron specific enolase (NSE) antibody. A Mann-Whitney U test was used in a statistical analysis. RESULTS: Most of the CgA-positive NE cells were localized between the acinar epithelial cells. The mean numbers of CgA-positive NE cells per acinus in the normal controls and the BPH groups were 1.67+/-0.78 and 4.45+/-2.54, respectively, and the difference was statistically significant (p<0.05). However, the mean number of CgA-positive NE cells in the androgen blockade group, was 4.93+/-2.17, which was similar to the BPH group. In a NSE immunoblotting study, a distinct band was observed in the BPH and androgen blockade groups, but the density of the band was higher in the androgen blockade group. CONCLUSIONS: Our results suggest that NE cells may be involved in the hyperplastic process of BPH. Inhibition of dihydrotestosterone, caused by the oral administration of the 5alpha-reductase inhibitor, failed to induce any significant change in the NE cells, probably due to the incomplete androgen blockade.

Urinary Profiles of the Endogenous Steroids in Pre-Menopausal Women with Uterine Leiomyoma / 대한불임학회지

Uterine leiomyoma is the most common solid pelvic tumor, occurring in 20~30% of women who are over 30 years of age1 and it accompanies with the symptoms such as uterine bleeding, dysmenorrhea, pain and the pressure on the urinary tract lead to blockage of the urinary tract. it some-times becomes a factor in sterility.2 Leiomyoma is a benign neoplasm that arises from uterine smooth muscle. it is hypothesized that leiomyoma originates from the somatic mutations in myometrial cells, resulting in progressive loss of growthregulation.3,4 Ovarian hormones are believed to stimulate the growth of leiomyoma because there is an increased incidence of leiomyoma after menarche and these tumors enlarge during pregnancy and regress after menopause. The growth of leiomyoma is variable among women with regular menstruation cycles and even among myoma nodules in the same uterus. One possible reason for this variation is thaovarian hormones, especially estrogen, stimulate individual myoma nodules by varying degree. 5 Therefore, therapeutic attempt based on over-come the state of hyperestrogenism have been tried. Treatment with competitive inhibitors of estrogen receptors (ER)6,7 or gonadotropin-releasing hor-mone agonist2,8,9 has been studied for those reasons. it was found that mean ER content was significantly greater in leiomyoma than in myometrium. 10~14 And ER content of the fibroid was reported to signifIcantly correlate wIth the myoma-shrinkage.15 But there were no consistent results as the concentration of estrogen in uterine leiomyoma.16,17 in this study, we determined the concentrations of urinary steroids, including estrogens as well as androgens, which are closely related to the estrogen biosynthesis, in premenopausal women with leiomyoma using Gas Chromatography-Mass Spectrometry (GC-MS). The urinary levels of the same endogenous steroids in age-matched healthy premenopausal women were also estimated by comparing urinary steroid levels between the two groups. From these results, we studied the effect of endogenous steroids and the metabolic changes in the leiomyoma, and we especially observed the difference in the estrogen level between the two groups to predict the role of estrogens in the prevention of and the therapy for leiomyoma

Comparative Study of Concentration of Isoflavones and Lignans in Plasma and Prostatic Tissues of Normal Control and Benign Prostatic Hyperplasia

Objective: Isoflavones and lignans are phytoestrogens that have recently gained interest as dietary factors related to prostatic diseases. However, no data on the concentrations in prostate tissue in humans is available. Therefore, the concentrations of isoflavones and lignans in plasma and prostatic tissues according to the prostate volume were compared to determine their possible effect on the benign prostatic growth. Methods: Fasting plasma and prostatic tissue specimens were acquired from 25 men over 50 years of age with similar normal dietary habits and no previous history of drug intake that could affect the isoflavones and lignans levels. The tissue was acquired either during a transurethral resection of the prostate in 15 patients with benign prostatic hyperplasia (BPH) with prostate volume over 40 ml or during a radical cystoprostatectomy in 10 patients with bladder cancer with a prostate volume < 25 ml, who were used as the controls. Quantitative analysis of the isoflavones, specifically equol, daidzein and genistein and lignans, particularly enterodiol and enterolactone, was performed by gas chromatography-mass spectrometry. Results: The mean prostatic concentrations of enterodiol, enterolactone, equol and daidzein in the BPH and the control groups were similar. However, the mean prostatic concentration of genistein was significantly lower in the BPH group than in the control group (65.43 +/- 17.05 vs 86.96 +/- 37.75 ng/ ml, respectively, p=0.032). The plasma concentration of isoflavones and lignans in the two groups were comparable. Conclusion: Isoflavones, but not lignans, have some influence the benign prostatic growth, and the prostatic concentration of genistein possibly has the closest association among them. More studies to further clarify the roles and mechanisms of isoflavone action on BPH including pharmacokinetic studies are recommended.

Development of Rat Prostatitis Model by Oral Administration of Isoflavone and Its Characteristics

Inflammation of the prostate can be induced experimentally in rats by the subcutaneous administration of estrogen. However, it is usually achieved at the price of some alteration in the sex steroid hormone balance and morphological changes in the prostate. In this study, a soy-extracted isoflavone mixture with weak estrogenic activity was administered orally in an attempt to induce prostatitis in a more physiologic way and to characterize the inflammation induced. A total of 36 male Sprague-Dawley rats, 8 weeks old, were divided into 2 groups. The control group was fed with only an AIN-76A diet containing no detectable phytoestrogen and the experimental group was fed with AIN-76A and a soy- extracted isoflavone mixture (genistein 60.0% and daidzein 19.6%), 300mg/kg body weight for 9 weeks. The sequential body weight and prostate weight at necropsy were measured. A histologic examination and histomorphometry assessed the changes in the prostate. The serum concentrations of testosterone and dihydrotestosterone were measured to estimate the effects on the androgen level. Intraprostatic concentrations of genistein and daidzein were measured by gas chromatography/ mass spectroscopy (GC/MS). While no sign of prostate inflammation was apparent in the control group, severe inflammatory changes in the stroma, increased epithelial detachment and inflammatory exudates within the glandular lumen of the dorsolateral prostate were observed in more than 80%(15/18) of the experimental group. However, there was no significant difference in the ventral prostate between the two groups. The daidzein and genistein concentrations in both the lateral and ventral prostates were significantly higher in the experimental group than in the control group where no isoflavone was detectable. In addition, the concentrations were much higher in the dorsolateral than in the ventral prostate. Although the body weight gain was not consistent in the experimental group, there were no significant differences in the prostate weight and serum androgen level between groups. In summary, when a soy-extracted genistein and daidzein-rich isoflavone mixture was administered orally into rats, prostatic inflammation with characteristic lobe specificity developed. The present method of inducing prostatitis seems to be a more physiologic than an estrogen-induced experimental model, and sequential pharmacokinetic studies might help in establishing this model as a more valuable tool in assisting future research in this field.

Effect of Endogenous Steroids on Human Papillary Thyroid Cancer: Alteration of Urinary Profile in Pre- and Post-operative Cases / 대한내분비외과학회지

No abstract available in English.

Correlation between Steroid Hormone Metabolites and Leiomyomas of Uterus / 대한불임학회지

OBJETIVE: To elucidate 1) whether there are any differences in the urine concentrations of steroid hormone metabolites between patients with leiomyoma and normal controls 2) the correlation between urinary profiles of steroid hormones and leiomyomas of the uterus according to their type, location, volume, and weight. MATERIALS OF METHODS: The study population consisted of 37 premenopausal patients with uterine leiomyoma and the control group consisted of 25 premenopausal normal volunteer women without uterine leiomyoma. Confirmation of the existence of uterine leiomyoma was done by ultrasonography and histopathological examination after surgery. The volume of the leiomyoma was estimated by trans-abdominal and/or trans-vaginal ultrasonography. The Leiomyomas were divided into 3 types (subserosal, intramural and submucosal). Seventeen patients had subserosal type of leiomyoma, 10 with the intramural type and 10 with the submucosal type. The locations of the leiomyoma were also divided into 3 groups (fundus, body and isthmus). Seventeen patients showed a fundus location, 10 in body, and 10 in isthmus. We compared urinary profiles of the endogenous steroids between patients with leiomyomas and normal controls, and also investigated the relationship between urinary profiles of the endogenous steroids and leiomyomas according to their type, location, volume and weight by using highly sensitive Gas Chromatography-Mass Spectrometry (GC-MS) system. RESULTS: The mean ages of the patients with leiomyomas and the control group were 43.1+/-5.6 and 40.6+/-7.2 years, the weights were 63.4+/-7.3 and 59.4+/-8.1 kg, and their heights were 155.4+/-4.8 and 159.3+/-4.8 cm respectively. Seventeen patients had subserosal, 10 had intramural, and 10 had submucosal leiomyomas. There were 17 patients with leiomyoma located in fundus, 10 in body and 10 in isthmus. 17beta-estradiol, 5-AT, 11-keto ET, 11beta-hydroxy An, 11beta-hydroxy Et, THS, THA, THE, alpha-cortolone, alpha-cortol, beta-cortol, 11beta-OH Et/11beta-OH An and E2/E1 were significantly increased in patients with leiomyoma than in the control group. 17beta-estradiol was significantly increased in the intramural and the submucosal types than in the subserosal type. There was no significant difference in the concentrations of urinary steroids according to the locations of leiomyomas. There was no significant relationship between the concentration of urinary steroids and the volume of the leiomyomas. 17beta-estradiol significantly decreased as the weight of uterus increased (r=-0.322, p=0.04). CONCLUSION: The concentrations of steroid hormone metabolites were generally increased in patients with leiomyoma but were not significantly related to the volume and weight of the leiomyomas. Our study suggests that steroid hormones may be involved in the initiation of leiomyomas but may not be involved in their progression. In addition, the concentrations of steroid hormone metabolites are not related to the leiomyoma type and location.

Reduction of Central Dopamine Release in Hyperprolactinemia / 대한내분비학회지

Background: Prolactin(PRL) secretion is tonically inhibited by doparnine that originates from the hypothalamic tuberoinfundibular tract and reaches the lactotroph via the hypophyseal portal vessel. Hyperprolactinemia associated with oligomenorrhea-amenorrhea, galactorrhea and/or infertility is mainly due to PRL-secreting pituitary adenoma(PA). The diagnosis of idiopathic hyperprolac- tinemia(IHP) is made, when hyperprolactinemia is sustained and all causes of hyperprolactinemia are excluded without radiological abnormality. It is not known, whether IHP and PA are two distinct entities or two subsequent phases of the same disease. The etiology of both disorders remains unresolved. We investigated that PRL hypersecretion in patients with IHP and PA may be the result of a defect in the central nervous system(CNS)-dopamine release, and that there may be some differences in pathogenesis of both diseases. Methods: We measured 24 hour-urinary dopamine, norepinephrine, epinephrine, and serum and 24 hour-urinary VMA(vanillyl rnandelic acid), HVA(homovanilic acid), DOPAC(3,4-dihydroxy phenylaceticacid), MHPG(3-methoxy 4-hydroxy phenylglycol) in 10 normal controls, 9 patients with IHP, and 17 patients with PA in the early follicular phase. Results: Urinary HVA and DOPAC concentrations, the major metabolites of CNS dopaminergic activity, were signficantly lower in both patients with IHP and PA compared with those in normal controls(p 0.05), whereas they were not different in both disease groups. Dopamine, norepine-phrine, epinephrine, MHPG concentrations were similar to those of the normal controls. Although VMA concentrations of both disease groups were significantly higher than those of normal controls, all of them were within normal range. Conelusion: Although our data are unable to establish the precise biochemical defect responsible for central dopamine deficiency in pathogensis of IHP and PA, we can support the presence of a pathological reduction of brain dopamine activity in IHP and PA.

Urinary Polyamine Profiles and Benign Prostatic Hyperplasia / 대한비뇨기과학회지

Polyamines are non-specific marker of cellular proliferation in many malignant tumors, and it is also increase in certain benign conditions. We measured the urinary polyamines to investigate the possibility as a marker of abnormal prostate growth and the correlation with various clinical parameters. Urinary polyamine concentrations in 27 cases of symptomatic benign prostatic hyperplasia (BPH) were compared with those in 32 cases of age matched normal controls. Urinary concentration of polyamine profiles were quantitatively determined by Gas Chromatography/Nitrogen Phosphorus Detector and they were calculated by the correction of gram creatinine. The concentrations of N-acetyl putrescine, N-acetyl cadaverine, spermidine(spd), N1-acetyl spermidine, N8-acetyl spermidine, and spermine(spm) showed significant increase in BPH compared with normal control(all p<0.05). Level of serum prostate specific antigen(PSA) in BPH patients was negatively correlated with the concentration of urinary spermidine(p=0.049). And the ratio of spm/spd correlated with the level of prostate volume(p=0.046). No significant correlations was found between other clinical parameters such as age, level of hemoglobin or erythrocyte count with polyamine profiles concentration. These data suggested that urinary concentration of polyamines in BPH are elevated compared with those in normal control. Altered regulation of the biosynthesis and metabolism of spermidine and spermine may be involved in BPH.

Quantitative analysis of endogenous steroids in human urine by using gas chromatography-mass spectrometry / 대한내분비학회지

No abstract available.