Urine Of Non-pregnant Women Research Articles

16S rRNA gene-based metagenomic analysis reveals differences in bacteria-derived extracellular vesicles in the urine of pregnant and non-pregnant women.

Recent evidence has indicated that bacteria-derived extracellular vesicles (EVs) are important for host–microbe communication. The aims of the present study were to evaluate whether bacteria-derived EVs are excreted via the urinary tract and to compare the composition of bacteria-derived EVs in the urine of pregnant and non-pregnant women. Seventy-three non-pregnant and seventy-four pregnant women were enrolled from Dankook University and Ewha Womans University hospitals. DNA was extracted from urine EVs after EV isolation using the differential centrifugation method. 16S ribosomal RNA (16S rRNA) gene sequencing was performed using high-throughput 454 pyrosequencing after amplification of the V1–V3 region of the 16S rDNA. The composition of 13 taxa differed significantly between the pregnant and non-pregnant women. At the genus level, Bacillus spp. EVs were more significantly enriched in the urine of the pregnant women than in that of the non-pregnant women (45.61% vs 0.12%, respectively). However, Pseudomonas spp. EVs were more dominant in non-pregnant women than in pregnant women (13.2% vs 4.09%, respectively). Regarding the compositional difference between pregnant women with normal and preterm delivery, EVs derived from Ureaplasma spp. and the family Veillonellaceae (including Megasphaera spp.) were more abundant in the urine of preterm-delivered women than in that of women with normal deliveries. Taken together, these data showed that Bacillus spp. EVs predominate in the urine of pregnant women, whereas Pseudomonas spp. EVs predominate in the urine of non-pregnant women; this suggests that Bacillus spp. EVs might have an important role in the maintenance of pregnancy.

Structural and immunosuppression study for bioactivities of Tamm-Horsfall glycoprotein and uromodulin

IntroductionTamm-Horsfall glycoprotein (THP) and uromodulin are the most abundant glycoproteins in the urine of non-pregnant women and pregnant women, respectively. Today, the structural profile and biological activities of these glycoproteins are controversial. ObjectivesTo investigate the glycoforms and immunosuppressive activities of THP and Uromodulin MethodsUromodulin were isolated from diatomaceous earth filtration, N- and O-glycans were then analysed with mass spectrometer MALDI-TOF/TOF. MTS viability, cytokine and immunophenotyping assays were carried out to investigate the immunomodulatory activities of both THP and uromodulin Results & DiscussionMALDI-TOF/TOF analysis showed that both THP and uromodulin expressed high mannose and complex-type N-glycan carrying sialic acid residues, Sda, LacNAc and LacdiNAc sequence as the capping antennae. Uromodulin tends to express multiple degree of sialylation, in which up to four sialic acid residues were expressed on its tetra-antennary Nglycans. Core 1 and core 2 type O-glycans were observed in THP and uromodulin, with THP expressing more O-glycans. MTS viability assay showed that 125μg/ml of uromodulin and 250μg/ml of THP significantly reduced the PHA-activated PBMC cell viability, to 85.64 ± 5.9 > % and 84.59 ±2.94%, respectively. Uromodulin was 2-fold more active in suppressing PBMC cell viability. Immunophenotyping analysis showed that CD4+ T helper cells, CD8+ cytotoxic T cells, CD56+ NK cells and CD14+ monocytes were suppressed by THP. It induced pro-inflammatory cytokines such as IL-1β, TNF and Th1 cytokine IFN-γ. On the other hand, uromodulin had slightly greater effect in suppressing CD4+ T helper cells and CD8+ cytotoxic T cells. It only induced IL-1β and suppressed both Th1 cytokine IFN-γ and Th2 cytokine IL-10. Hence, uromodulin is more immunosuppressive than THP because it suppressed CD4+ and CD8+ T cells, stimulated IL-1β only and withheld the TNF secretion. ConclusionsGlycosylation changes as observed in THP are suggested to contribute to their differential immunosuppressive property. Developing

Application of β-cyclodextrin for the analysis of estrogenic steroids in human urine by high-performance liquid chromatography

A procedure is described for simultaneous determination of estriol, estrone and 17β-estradiol in human urine. After acid hydrolysis of the sulphate conjugates, the estrogens were extracted into diethyl ether. The ether extracts were concentrated and applied directly to an HPLC column. Using a 25 cm C-18 column and acetonitrile-water modified by the addition of β-cyclodextrin as mobile phase, the separation of estriol, estradiol and estrone was achieved within 20 minutes. The extraction of estrogens from the biological matrix is excellent. Estrogens were detected using a UV-detector (280 nm) or a spectrofluorimetric detector (λexc=280nm, λem=312 nm). The latter detection system has been used for the determination of estrogens in the urine of non-pregnant women. The procedure is simple and can be used in clinical practice.

Measurement of urinary estriol glucuronides during the menstrual cycle by high-performance liquid chromatography

A simple high-performance liquid chromatographic (HPLC) method for measuring estriol-3-glucuronide (E 3-3-G) and estril-16-glucuronide (E 3-16-G) in the urine of non-pregnant women is described. Estriol conjugates were extracted from 4 ml of urine with a small cartridge of graphitized carbon black (Carbopack B). After washing, E 3-3-G and E 3-16-G were desorbed separately by a two-step elution system. After solvent removal, the two glucuronides were quantified by isocratic ion-suppression HPLC with fluorimetric detection. The analytical recovery of the two estriol metabolites was about 95%. The detection limit of the method was 0.6 ng/ml for both analytes in urine, which is well below the concentrations of clinical interest, and the method is not susceptible to substantial interferences. Data relative to urinary levels of E 3-3-G and E 3-16-G measured by this method on a daily basis in early morning samples from the nine women during their menstrual cycles were compared with those reported in the literature and obtained radioimmunoassay techniques. Moreover, the potential use of defined changes in the concentrations of the two conjugates for predicting the fertile period of women was assessed.

Identification of a New Steroid: 3β,21-Dihydroxy-5α-pregnan-20-one in the Urine of Human Pregnancy

A new corticosteroid was isolated from the urine of women in the third trimester of pregnancy and identified as 3β,2l-dihydroxy-5α-pregnan-20-one. This steroid was obtained after hydrolysis and was isolated in different chromatographic systems. The identification was based on its chromatographic migration in different systems before and after acetylation, by the identification of the reduced product and by gas chromatography-mass spectrometry. The urinary excretion values per 24 hr are 650–1080 μg at 7–8½ months of gestation. Only traces of this steroid were found in the urine of nonpregnant women and it was not detected in the urine of men.

Isolation and identification of progesterone from urine of nonpregnant women.

Journal Article Letters to the Editor: Isolation and Identification of Progesterone from Urine of Nonpregnant Women Get access H. J. VAN DER MOLEN, H. J. VAN DER MOLEN 1Department of Obstetrics and Gynecology, University Hospital Utrecht; Department of Biochemistry, School of Medicine Rotterdam, The Netherlands Search for other works by this author on: Oxford Academic Google Scholar C. CORPECHOT C. CORPECHOT 1Department of Obstetrics and Gynecology, University Hospital Utrecht; Department of Biochemistry, School of Medicine Rotterdam, The Netherlands Search for other works by this author on: Oxford Academic Google Scholar The Journal of Clinical Endocrinology & Metabolism, Volume 28, Issue 9, 1 September 1968, Pages 1361–1366, https://doi.org/10.1210/jcem-28-9-1361 Published: 01 September 1968 Article history Received: 27 December 1967 Accepted: 17 May 1968 Published: 01 September 1968

EINE CHEMISCHE METHODE ZUR BESTIMMUNG VON 16-EPI-ÖSTRIOL IM URIN DES MENSCHEN

ABSTRACT A method for the chemical determination of 16-epi-oestriol in the urine of nonpregnant women with a qualitative sensitivity of less than 0.5 μg/24 h is described. The separation of 16-epi-oestriol and oestriol is accomplished by converting 16-epi-oestriol into its acetonide, a reaction which is stereoselective for cis-glycols and therefore not undergone by oestriol as a trans-glycol. Following partition between chloroform and aqueous alkali, the acetonide of 16-epi-oestriol is completely separated with the organic layer whereas oestriol as a strong phenol remains in the alkaline phase. 16-epi-oestriol is chromatographed on alumina as the acetonide and determined as a Kober chromogen. This procedure can easily be incorporated into the method of Brown et al. (1957 b) thus making possible the simultaneous routine assay of oestradiol-17β, oestrone, oestriol and 16-epi-oestriol from one sample of urine. The specificity of the method was established by separation of 16-epi-oestriol from nonpregnancy urine as the acetonide, hydrolysis of the acetonide by phosphoric acid, isolation of the free compound by microsublimation and identification by micro melting point, colour reactions and chromatography. The accuracy of the method is given by a mean recovery of 64% for pure crystalline 16-epi-oestriol when added to hydrolysed urine in 5–10 μg amounts. The precision is given by s = 0.24 μg/24 h. For the duplicate determination of 16-epi-oestriol the qualitative sensitivity is 0.44 μg/24 h, the maximum percentage error being ± 100% The quantitative sensitivity (±25% error) is 1.7 μg/24 h.

AN EVALUATION OF THE FRANK METHOD FOR THE DETERMINATION OF PROLAN (GONADOTROPIC PRINCIPLE) IN THE URINE OF NONPREGNANT WOMEN1

For some time we have been seeking a quantitative method for determining prolan in the urine of nonpregnant women when it is present in amounts of less than 66 r.u. per liter. The Aschheim-Zondek alcohol precipitation method (1), which employs 60 cc. of morning urine, does very well when amounts of prolan in the urine are in excess of 66 r.u. We found that many women suffering from vasomotor disturbances of the menopause did not have sufficient prolan in 60 cc. of urine to give a positive test; yet we felt that their urinary prolan must be in excess of normal. It is not so important to have a method for the detection of excess prolan in the urine of women who are able to describe typical vasomotor waves, but in cases of those suffering from some bizarre syndrome or psychosis, or those whose

A Source of Error in Gonadotropic Hormone Determihations.

This report is presented in an effort to show that determinations for gonadotropic hormone content of the urine are frequently erroneous due to the precipitation of contaminating amounts of fat-insoluble or combined estrogen.Concerning urinary gonadotropins there is no doubt that pregnant women excrete a substance capable of producing corpora lutea in the ovaries of normal infantile animals. Some women in the menopause excrete a gonadotropic hormone capable of provoking follicle growth in the ovaries of hypophysectomized rats or mice. A gonadotropic hormone prepared from the pituitary and capable of interstitial cell stimulation has been described, but its detection if present in the urine, depends also upon the use of hypophysectomized animals. For studying the urine of non-pregnant women most workers have resorted to the use of intact animals and depend upon more cursory and unreliable criteria for the estimation of gonadotropic hormone. Speaking collectively, their criteria have been the appearance, in...

Method of Preparing Mice for Quantitative Determination of Urinary Estrogen.

The methods of biological assay for urinary estrogen in use at the present time are dependent upon the production of estrus in a castrated mouse or rat. Various modifications of the original Allen-Doisy test have been used, such variations being single or multiple injections of either aqueous, glycerol, or oily extracts into castrated or immature normal mice or rats. Methods utilizing male animals and fish have not as yet proved of any great value for quantitative purposes. Chemical methods for the determination of urinary estrogen have been proposed by Kober, Cohen and Marrian, David, Cartland et al., Schmulovitz and Wylie,, and Pincus et al. Chemical tests based on color changes in the test tube and comparison with standards are preferable to biological tests. The disadvantage of chemical tests is chiefly their inapplicability to the estimation of estrogens in the urine of non-pregnant women. For this purpose we must, for the present, still resort to biological means. A biological test for urinary estrogen must be for the biological activity of all the estrogenic substances present in the urinary extract. The test cannot be specific for, or differentiate between, different forms of the hormone. The advantage of the biological method is its ability to demonstrate very small amounts of estrogenic hormone in the presence of inactive impurities. The disadvantages are the troublesome time consuming procedures and the necessity of maintaining a supply of test animals. The greatest disadvantage is the source of error and variation in methods. More than a 4000% variation can he demonstrated in the procedures now used. This paper is presented in an ctiort to decrease the large percent of error and variation. Our test animal I)!- choicc has ken the castrated adult female mouse. just as chemically piire hornic me ior assay is tested against a knon-n ctantlartl hormone.

Colorimetric Estimation of strin in the Urine of Non-Pregnant Women

LAST year we reported the details of a method by which it is possible to determine the “strone and “striol content of human pregnancy urine colorimetrically with a reasonable degree of accuracy1. Since the publication of our paper we have received numerous inquiries as to whether the method can be used for the determination of the “strin content of urine of non-pregnant women. As this question seems to be one of some general interest, and since in our paper we made no mention of the possibility of using our method for this purpose, we wish to take this opportunity of mentioning briefly our views on the matter.