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Sample records for plasma progesterone determinations

  1. Effects of ergocryptine on plasma prolactin, luteinizing hormone, and progesterone in the periparturient sow.

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    Whitacre, M D; Threlfall, W R

    1981-09-01

    Effects of ergocryptine (ERG) on the periparturient sow were assessed by measuring changes in prolactin, luteinizing hormone, and progesterone in plasma. Hormones in plasma were determined on samples collected daily from 21 days before parturition until postpartum day (PPD) 21. Administration of ERG completely blocked the initiation of lactation and prevented the surge in prolactin that occurred just before parturition, as compared with effects in control sows. The gestation period was shorter (P less than 0.01) in ERG-treated sows compared with the gestation period in controls. Although the number of pigs farrowed was not different between treated and control sows, the ERG-treated sows did not raise any pigs to PPD 21. Luteinizing hormone concentrations were higher (P less than 0.01) from 10 days before parturition until PPD 21, but plasma progesterone concentrations were not different in treated sows when compared with concentrations in control sows.

  2. A comparative study of clomiphene and epimestrol on plasma progesterone, CBG, TBG and SHBG, and salivary progesterone levels.

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    Kicovic, P M; Luisi, M; Cortés-Prieto, J; Franchi, F

    1982-01-01

    The plasma levels of CBG, TBG, SHBG and progesterone, and salivary progesterone levels were measured in eight young ovulatory volunteers. After the control cycle four subjects received 50 mg/day of Clomiphene from days 5 to 9 of the first treatment cycle, and 10 mg/day of Epimestrol from days 5 to 15 of the second treatment cycle. The other four subjects received the treatments in reverse order. Between the two treatments there was one cycle without medication as a "wash-out" period. Plasma and saliva samples were obtained on days +6, +8 and +10 (day of LH peak was denoted by 0), always between 08.00 and 09.00 h. Statistical evaluation was done by means of an analysis of variance (ANOVA), and correlation coefficients were also calculated. Evaluation of data on effects of Clomiphene and Epimestrol on the plasma levels of CBG and salivary progesterone showed that Clomiphene induced a highly significant rise (p less than 0.001) in the CBG levels and decrease (p less than 0.001) in salivary progesterone levels, while no changes were seen following administration of Epimestrol. Both Clomiphene and Epimestrol treatments led towards higher plasma progesterone levels, those following Clomiphene administration being higher. Neither treatment induced significant changes in TBG or SHBG levels. It is concluded that Clomiphene induces significant elevations of CBG and decrease in salivary progesterone, which is thought to reflect the free progesterone fraction and may have significance in relation to a discrepancy between the ovulation and pregnancy rates following Clomiphene therapy.

  3. CORRELATION OF PROGESTERONE AND CORTISOL PLASMA LEVELS BETWEEN OVULATED AND NON-OVULATED ETTAWA CROSSBREDS DOES

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    Pudji Astuti

    2008-03-01

    Full Text Available This experiment was conducted to determine the correlation of progesterone and cortisol levels in plasma between ovulated and non-ovulated Crossbreds Ettawa does. Eight does were used in this experiment and they were divided equally into 2 groups, i.e. group I consisting of 4 normal ovulated does with the average body weight of 37.5 ± 3.109 kg and group II consisting of 4 non-ovulated does with the average body weight of 28 ± 2.160 kg as group II. The estrus cycles of all does were synchronized using intravaginal device of controlled internal drug release (CIDR accompanied by intra-muscular injection of prostaglandin F2 (PGF2 alpha. Immediately after the onset of estrus, blood samples were collected from jugular every 3 and 6 hours for 72 hours after onset of estrus. The concentration of cortisol and progesterone in plasma was assayed by enzyme linked immunosorbent assay (ELISA. The results showed that the average concentrations of cortisol in ovulated does (90.89±26.22 ng/mL was higher than in non-ovulated does (42.70±37.18 ng/mL. Similarly, the concentrations of progesterone in ovulated does (0.098±0.0423 ng/mL was higher than in non-ovulated does (0.093±0.056 ng/mL. It was evident that the change in progesterone level was closely associated with the change in cortisol level in plasma.

  4. Setting-up and validation of two radioimmunoassay methods for determination of plasma progesterone concentration in mares, cows and rats; Padronizacao e validacao de dois metodos de radioimunoensaio (RIE) para dosagem da progesterona (P{sub 4}) no plasma de equinos, bovinos e ratos

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    Rosa e Silva, A.A.M.; Caldas, M.C.S.; Campos, L.M.A. [Sao Paulo Univ., Ribeirao Preto, SP (Brazil). Fac. de Medicina. Dept. de Fisiologia; Gradela, A. [UNESP, Jaboticabal, SP (Brazil). Faculdade de Ciencias Agrarias e Veterinarias

    1993-06-01

    Two reliable radioimmunoassay (RIA) methods which permits the measurement of progesterone (P{sub 4}) in plasma of equine, bovine and rats are described. After extraction of plasma with diethylic ether the RIA methods were performed. The first one utilizes {sup 125} I progesterone (double antibody method) and the other 1,2,6,7,16, 17 {sup 3} H progesterone (adsorption in charcoal/dextran). Both two methods were suitable in the valuation of plasma P{sub 4} concentration in different physiological reproductive conditions. The method of the double antibody showed higher sensibility beyond to be less expensive than the other method. Despite it, the two RIA methods were much less expensive than available commercial Kits in the market. (author) 14 refs., 2 figs., 1 tab.

  5. Response of plasma cortisol and progesterone after ACTH challenge in ovariectomized lactating dairy cows.

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    Yoshida, Chikako; Nakao, Toshihiko

    2005-02-01

    Shortened and weakened estrous expressions could be one of the causes of poor heat detection rate. Non-specific acute stresses are assumed to depress expression of estrus by an increase of plasma progesterone which may originate from the adrenal cortex. The objective of the present study was to examine whether the adrenal cortex can secrete significant amounts of progesterone in response to exogenous adrenocorticotropic hormone (ACTH) in lactating cows. Four cows had estrus synchronized and were ovariectomized in the luteal phase. The cows were given 25 IU ACTH through an indwelling catheter 5 h after catheterization. Blood samples were collected at an interval of 30 min. In 3 of the 4 cows, plasma progesterone concentrations increased significantly 0.5-1.5 h after the first ACTH challenge with a mean peak value of 4.2 +/- 0.4 (S.D.) ng/ml. A similar response was also observed after the second ACTH challenge. Peak plasma progesterone concentrations in the 3 cows after first ACTH challenge were comparable with the progesterone values in the luteal phase of each cow. The results suggest that lactating cows have the capability to secrete a significant amount of progesterone from the adrenal cortex.

  6. Longitudinal studies of plasma aldosterone, corticosterone, deoxycorticosterone, progesterone, 17-hydroxyprogesterone, cortisol, and cortisone determined simultaneously in mother and child at birth and during the early neonatal period. I. Spontaneous delivery.

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    Sippell, W G; Becker, H; Versmold, H T; Bidlingmaier, F; Knorr, D

    1978-06-01

    In order to obtain the still lacking reference data of individual plasma steroids in the immediate postnatal period needed for the assessment of adrenocortical function in various neonatal maladaptation syndromes, aldosterone (A), corticosterone, deoxycorticosterone (DOC), progesterone (P), 17-hydroxyprogesterone (17-OHP), cortisol, and cortisone were simultaneously followed in the same human newborn in a single 250-500 microliters peripheral plasma sample obtained at constant times during the first week of life using a mechanized Sephadex LH-20 multicolumn chromatography and standardized RIAs. Mean concentrations in 12 spontaneously delivered full term newborns of either sex and in paired umbilical (UV) and peripheral maternal (MV) venous plasma are given in the table. Besides significant maternoumbilical gradients in each steroid, DOC, P, 17-OHP, and cortisone, originating predominantly from the fetoplacental unit, disappear rapidly with steadily increasing half-lives. A, corticosterone, and cortisol, however, remain elevated in comparison with later infancy, with the exception of a marked "glucocorticoid dip" in cortisol and corticosterone levels between 2 and 12 h after birth.

  7. Evaluation of extraction methods for progesterone determination in rabbit (Oryctolagus cuniculus) feces by radioimmunoassay

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    Korndoerfer, Clotilde Maria; Meirelles, Cyro Ferreira; Bueno, Ives Claudio da Silva; Abdalla, Adibe Luiz [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Secao de Ciencias Animais]. E-mail: cfmeirel@esalq.usp.br

    1998-07-01

    The purpose of this study was to find a practical procedure for the extraction of progesterone (P{sub 4}) from feces and to determine if the P4 plasma profiles during pregnancy were reflected in total fecal P4 of pregnant rabbits. The rabbit was used as model for the techniques. Plasma and feces were collected from 11 rabbits during a period of 42 days. Three different methods of P4 extraction were used. The total P4 was measured by solid-phase radioimmunoassay (RIA) with {sup 125} I-P4 as the tracer. Results suggested that it was possible to extract total P4 from rabbit feces with methanol and petroleum ether. Plasma and fecal P4 profiles were compared for both pregnant and ovariectomized rabbits. It was possible to differentiate total P4 extracted from day two through 28 after breeding (p<0.01). (author)

  8. Corticotropin-releasing hormone and progesterone plasma levels association with the onset and progression of labor.

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    Stamatelou, F; Deligeoroglou, E; Vrachnis, N; Iliodromiti, S; Iliodromiti, Z; Sifakis, S; Farmakides, G; Creatsas, G

    2013-01-01

    PURPOSE OF LNVESTIGATION: To examine the relationship between maternal plasma progesterone along with corticotropin- releasing hormone (CRH) plasma levels and the progression of labor. Maternal serum CRH and progesterone were measured during the latent phase of labor, active labor, and 24 hours postpartum in women who went into spontaneous labor and delivered vaginally at term. Progesterone (P) levels in women delivered by an elective cesarean section at term were also measured as baseline. Mean maternal plasma P was 18% higher in the active phase than in the latent phase of labor (p labor (p labor progresses, P and CRH increase and subsequently decrease precipitously in the immediate postpartal period. P levels tend to drop in women who are in early labor compared with non-laboring full-term women.

  9. Plasma prolactin, progesterone and oestradiol-17β concentrations around parturition in the pig

    NARCIS (Netherlands)

    Taverne, M.; Willemse, A.H.; Dieleman, S.J.; Bevers, M.

    1979-01-01

    Plasma concentrations of prolactin, progesterone and oestradiol-17β were measured by radioimmunoassay in samples taken from 2–15 days before until 1–4 days after spontaneous parturition in four sows and in one sow around prostaglandin F2α-induced parturition. Between Days −15 and −2 (Day 0 = partur

  10. Plasma prolactin, progesterone and oestradiol-17β concentrations around parturition in the pig

    NARCIS (Netherlands)

    Taverne, M.; Willemse, A.H.; Dieleman, S.J.; Bevers, M.

    Plasma concentrations of prolactin, progesterone and oestradiol-17β were measured by radioimmunoassay in samples taken from 2–15 days before until 1–4 days after spontaneous parturition in four sows and in one sow around prostaglandin F2α-induced parturition. Between Days −15 and −2 (Day 0 =

  11. Investigation of pig sperm plasma membrane reorganization using progesterone-albumin-fluorescein probes

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    Alfredo Medrano; Paul F Watson; William V Holt

    2012-01-01

    Objective:To relate semen susceptibility in cooling protocols to sperm plasma membrane properties.Methods:A series of experiments was performed using the fluorescent markers, progesterone-BSA-FITC andBSA-FITC.Results:These experiments indicated that both progesterone-BSA-FITC andBSA-FITC bound to specific sperm plasma membrane domains, thus producing four different binding patterns, revealing probable changes in membrane organization during capacitation and during cooling.Those patterns seem to make a sequence progressing from non-capacitated status to capacitated status.The proportion of each pattern was different during incubation than during cooling, showing the latter had a higher proportion of dead sperm than the former.Conclusions:At this stage, the association of sperm plasma membrane alterations was revealed byBSA-FITC probes and cryosensitivity remains unclear.

  12. Plasma urea nitrogen and progesterone concentrations and follicular dynamics in ewes fed proteins of different degradability

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    Gustavo Bianchi Lazarin

    2012-07-01

    Full Text Available The effects of overfeeding with protein of different degradability on body condition, plasma urea nitrogen and progesterone concentrations, ovulation number and follicular dynamics were assessed in Santa Ines ewes. Twelve ewes were assigned to a randomized block design according to body weight and received overfeeding with soybean meal or with corn gluten meal or maintenance diet for 28 days before ovulation and during the next estrous cycle. Blood samples were taken on days 7, 14, 21, and 28 after the beginning of treatments for analysis of plasma urea nitrogen and on days 3, 6, 9, 12, and 15 into the estrous cycle for analysis of plasma urea nitrogen and progesterone. Follicular dynamics was monitored daily by ultrasound during one estrous cycle. Dry matter and crude protein intake, weight gain, plasma urea nitrogen concentration before ovulation, number of ovulations, diameter of the largest follicle of the 1st and of the 2nd waves and the growth rate of the largest follicle of the 1st wave were higher in the ewes that received overfeeding. The growth rate of the largest follicle of the 3rd wave was higher in the ewes fed maintenance diet. The back fat thickness, plasma urea nitrogen before ovulation and progesterone concentrations, diameter of the largest follicle of the 2nd wave and growth rate of the largest follicle of the 3rd wave were higher in ewes that received overfeeding with soybean meal. The growth rate of the largest follicle of the 1st wave was higher in ewes that received overfeeding with corn gluten meal. Overfeeding with protein-rich feeds may increase the ovulation number and with soybean meal, it may be effective in increasing plasma progesterone concentration in ewes.

  13. Effect of progesterone on renal sodium handling in man: relation to aldosterone excretion and plasma renin activity.

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    Oparil, S; Ehrlich, E N; Lindheimer, M D

    1975-08-01

    1. The effect of progesterone on renal haemodynamics and intrarenal sodium handing was evaluated in thirteen normal men on a constant diet. Clearances were measured during maximal water diuresis and again 4-7 days later, this time 3 h after progesterone was given intramuscularly. Seven additional studies were performed 3 days after progesterone administration. Another four tests were performed on volunteers who had manifested renal 'escape' from the sodium-retaining effect of deoxycorticosterone acetate. 2. In acute progesterone studies glomerular filtration rate was unchanged, whereas effective renal plasma flow increased, so that filtration fraction decreased significantly. A similar in crease in urinary sodium occurred whether subjects received a low or high sodium diet. Indices which related to the distal delivery of filtrate (fractional urine flow and the sum of fractional free water and sodium clearances) increased significantly in both groups. The progesterone-induced increase in sodium excretion was not related to changes in plasma renin activity, renin substrate or urinary aldosterone. After 3 days of progesterone, the increase of sodium excretion was less than in the acute studies and urinary aldosterone increased tow- to four-fold. Progesterone failed to produce an acute increse in urinary sodium in subjects hyperexpanded by administration of exogenous mineralocorticoids. 3. Results suggest that the acute natriuretic action of progesterone is in part independent of aldosterone inhibition and that progesterone may inhibit sodium reabsorption at proximal as well as distal sites in the nephron.

  14. Progesterone stimulates adipocyte determination and differentiation 1/sterol regulatory element-binding protein 1c gene expression. potential mechanism for the lipogenic effect of progesterone in adipose tissue.

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    Lacasa, D; Le Liepvre, X; Ferre, P; Dugail, I

    2001-04-13

    Fatty acid synthase (FAS), a nutritionally regulated lipogenic enzyme, is transcriptionally controlled by ADD1/SREBP1c (adipocyte determination and differentiation 1/sterol regulatory element-binding protein 1c), through insulin-mediated stimulation of ADD1/SREBP1c expression. Progesterone exerts lipogenic effects on adipocytes, and FAS is highly induced in breast tumor cell lines upon progesterone treatment. We show here that progesterone up-regulates ADD1/SREBP1c expression in the MCF7 breast cancer cell line and the primary cultured preadipocyte from rat parametrial adipose tissue. In MCF7, progesterone induced ADD1/SREBP1c and Metallothionein II (a well known progesterone-regulated gene) mRNAs, with comparable potency. In preadipocytes, progesterone increased ADD1/SREBP1c mRNA dose-dependently, but not SREBP1a or SREBP2. Run-on experiments demonstrated that progesterone action on ADD1/SREBP1c was primarily at the transcriptional level. The membrane-bound and mature nuclear forms of ADD1/SREBP1 protein accumulated in preadipocytes cultured with progesterone, and FAS induction could be abolished by adenovirus-mediated overexpression of a dominant negative form of ADD1/SREBP1 in these cells. Finally, in the presence of insulin, progesterone was unable to up-regulate ADD1/SREBP1c mRNA in preadipocytes, whereas its effect was restored after 24 h of insulin deprivation. Together these results demonstrate that ADD1/SREBP1c is controlled by progesterone, which, like insulin, acts by increasing ADD1/SREBP1c gene transcription. This provides a potential mechanism for the lipogenic actions of progesterone on adipose tissue.

  15. [Simultaneous study of plasma gonadotropins, estrogens, progesterone and 17-hydroxyprogesterone during the ovulatory cycle].

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    Roger, M; Veinante, A; Soldat, M C; Tardy, J; Tribondeau, E; Scholler, R

    1975-09-20

    Plasma levels of gonadotrophins, estrone, estradiol, 17-hydroxyprogesterone and progesterone were studied throughout an ovulatory cycle in 11 normal women. For each of these hormones, the initiation of a secretory cycle occured at a different time: on the 10th day after mid-cycle surge for gonadotrophins, on the 3 rd day of menses for estradiol and somewhat later for estrone, on the 8th day of cycle for 17-hydroxyprogesterone and on the day of the LH surge for progesterone. Mean plasma levels and range were reported for every studied hormone. The mid-cycle surges of LH and FSH were found always coincident, after which their concentration decreased simultaneously; during the late follicular phase the increasing LH curve intersected the decreasing FSH curve. A peak of estradiol occured in every subject except one on the day before the mid-cycle surge of LH; a peak of plasma estrone occured either on the day of the LH peak or on the day before. For these two hormones the luteal phase range is always below the peak level. A peak of 17-hydroxyprogesterone occured in every subject on the same day than the mid-cycle LH peak, but the plasma level increased during the luteal phase up to higher levels. Progesterone concentration increased on the day of the LH surge, indicating the initiation of luteinization prior to ovulation.

  16. Plasma PGFM and progesterone concentrations, luteolysis moment and estrous cycle length in Nelore cows submitted to uterine biopsies

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    I. Martin

    2015-10-01

    Full Text Available ABSTRACTIn this study, endometrial samples were collected in 14 Nelore cows on days zero (ovulation, five, nine, thirteen and nineteen of the estrous cycle (biopsy group, and in 15 females these collections weren't performed (control group. Biopsies were done on the uterine horn endometrium contralateral to the ovary with corpus luteum. Blood samples were taken at -24, -16, -8, 0 +8, +16 and +24 hours in relation to progesterone drop (<1ng/mL, zero moment and evaluated for 13, 14-dihydro-15-keto prostaglandin F2-alpha (PGFM by radioimmunoassay (RIA. Plasma progesterone concentration was determined by RIA every 24 hours. Data were analyzed by ANOVA using the PROC GLM and MIXED of the SAS. The mean value for PGFM during the entire period evaluated was greater in the biopsy group. The mean concentration of PGFM at moment zero was not different between the groups; the mean concentration of PGFM was higher in the biopsy group before and after the drop in progesterone level. The maximum mean concentration observed was not different between the groups; however, the mean minimum concentration was different with high values in the biopsy group. Although the PGFM concentrations were higher in the biopsy group, the biopsy and control groups had similar length of estrous cycle showing that repeated endometrial biopsy on the side contralateral to the ovary with corpus luteum does not affect luteolysis and the length of the estrous cycle.

  17. Detection and identification of plasma progesterone metabolites in the female Florida manatee (Trichechus manatus latirostris) using GC/MS/MS.

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    Tripp, K M; Dubois, M; Delahaut, P; Verstegen, J P

    2009-08-01

    Florida manatees (Trichechus manatus latirostris) have relatively low peripheral concentrations of progesterone (P4). The objective of this study was to determine if these relatively low P4 concentrations are associated with a high ratio of progestin metabolites and to document metabolite concentrations from individual blood samples obtained from manatees during diestrus or pregnancy. Metabolites known to exist in elephants-terrestrial manatee relatives-were targeted. These included 5alpha-reduced progestins (5alpha-pregnane-3,20-dione [5alpha-DHP] and 3alpha-hydroxy-5alpha-pregnan-20-one [5alpha-P3-OH]) and 17alpha-hydroxyprogesterone (17alpha-OHP), which occurs in Asian elephants. An additional, inactive metabolite, 20alpha-hydroxyprogesterone (20alpha-OHP), indicative of P4 overproduction, was also targeted. Progesterone itself was the predominant progestin detected in pregnant and nonpregnant manatee plasma (n = 10) using gas chromatography-mass spectrometry with tandem quadrupole detectors (GC/MS/MS). Progesterone concentrations in pregnant females varied from early (moderate to high) through mid and late (low) pregnancy. Progesterone concentrations ranged from low to high in nonpregnant, nonlactating females. The most commonly detected metabolite was 5alpha-P3-OH (n = 7), which occurred in pregnant (lower limit of detection [LLOD] to high) and nonpregnant (trace to high) females. The 5alpha-DHP metabolite was also detected in pregnant (LLOD to moderate) and nonpregnant (low) females. The 17alpha-OHP metabolite was not detected in any tested female. The 20alpha-OHP metabolite was detected in one nonpregnant, nonlactating, captive female (LLOD). Metabolites were most prevalent during early pregnancy, concurrent with maximum P4 concentrations. Based on their concentrations in peripheral circulation, we inferred that these metabolites may have, opposite to elephants, a limited physiologic role during luteal, pregnant, and nonpregnant phases in the manatee.

  18. Effect of incremental levels of crude degummed canola oil on milk progesterone, plasma

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    John R. Otto

    2014-12-01

    Full Text Available Dietary supplementation of lactating cows with fat can alter the profiles of key reproductive hormones and boost postpartum energy balance. However, published data under Australian pasture-based dairy production conditions are scanty and inconsistent. Therefore, the objective of this study was to determine whether dietary inclusion of crude degummed canola oil (CDCO at incremental levels for eight-weeks will have significant influence on progesterone (P4, luteinizing hormone (LH and follicle stimulating hormone (FSH of primiparous Holstein–Friesian cows grazing pastures. We tested the hypothesis that postpartum supplementation of primiparous Holstein–Friesian cows with dietary CDCO in a pasture-based system will alter the concentrations of P4, LH and FSH reproductive hormones. A random allocation of twenty primiparous Holstein–Friesian cows into four treatment groups that consisted of a wheat-based pelleted basal diet with no supplemental CDCO (control, or a wheat-based pelleted basal diet with CDCO added at 25 ml/kg (low, 35 ml/kg (medium and 50 ml/kg (high was employed in an eight-week feeding trial after two weeks of adjustment. Supplementation levels of CDCO and week of data collection were significant sources of variation (P  0.05. It was apparent that cows in the high (0.459 ng/ml, medium (0.367 ng/ml and low (0.251 ng/ml levels of oil treatments had higher mean plasma FSH concentrations compared to the control (0.172 ng/ml cows. It was concluded that the current levels of CDCO can be used in pasture-based dairy systems to increase FSH, but not LH and P4.

  19. Serum progesterone

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    ... more commonly) Determine the risk of miscarriage or ectopic pregnancy early in pregnancy Normal Results Progesterone levels vary ... levels may be due to: Amenorrhea (no periods) Ectopic pregnancy Failure to ovulate Fetal death Possible miscarriage Alternative ...

  20. The association among cytochrome P450 3A, progesterone receptor polymorphisms, plasma 17-alpha hydroxyprogesterone caproate concentrations, and spontaneous preterm birth.

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    Bustos, Martha L; Caritis, Steve N; Jablonski, Kathleen A; Reddy, Uma M; Sorokin, Yoram; Manuck, Tracy; Varner, Michael W; Wapner, Ronald J; Iams, Jay D; Carpenter, Marshall W; Peaceman, Alan M; Mercer, Brian M; Sciscione, Anthony; Rouse, Dwight J; Ramin, Susan M

    2017-09-01

    Infants born preterm birth are the leading cause of mortality in children preterm birth by 33% in women with history of spontaneous preterm birth. We demonstrated previously that plasma concentrations of 17-alpha hydroxyprogesterone caproate vary widely among pregnant women and that women with 17-alpha hydroxyprogesterone caproate plasma concentrations in the lowest quartile had spontaneous preterm birth rates of 40% vs rates of 25% in those women with higher concentrations. Thus, plasma concentrations are an important factor in determining drug efficacy but the reason 17-alpha hydroxyprogesterone caproate plasma concentrations vary so much is unclear. Predominantly, 17-alpha hydroxyprogesterone caproate is metabolized by CYP3A4 and CYP3A5 enzymes. We sought to: (1) determine the relation between 17-alpha hydroxyprogesterone caproate plasma concentrations and single nucleotide polymorphisms in CYP3A4 and CYP3A5; (2) test the association between progesterone receptor single nucleotide polymorphisms and spontaneous preterm birth; and (3) test whether the association between plasma concentrations of 17-alpha hydroxyprogesterone caproate and spontaneous preterm birth varied by progesterone receptor single nucleotide polymorphisms. In this secondary analysis, we evaluated genetic polymorphism in 268 pregnant women treated with 17-alpha hydroxyprogesterone caproate, who participated in a placebo-controlled trial to evaluate the benefit of omega-3 supplementation in women with history of spontaneous preterm birth. Trough plasma concentrations of 17-alpha hydroxyprogesterone caproate were measured between 25-28 weeks of gestation after a minimum of 5 injections of 17-alpha hydroxyprogesterone caproate. We extracted DNA from maternal blood samples and genotyped the samples using TaqMan (Applied Biosystems, Foster City, CA) single nucleotide polymorphism genotyping assays for the following single nucleotide polymorphisms: CYP3A4*1B, CYP3A4*1G, CYP3A4*22, and CYP3A5*3; and rs

  1. [New method for the determination of estrogen and progesterone receptors in human breast cell lines].

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    Olea, N; Devleeschower, N; Leclercq, G

    1985-12-01

    A method for the determination of estrogen and progesterone receptor levels in human mammary cell lines (MCF-7, Cama-1, ZR-75-1, Evsa-T and HBL-100) is described. Cells cultured as monolayers were incubated with the tritiated steroids, [3H]-17 beta-Estradiol or [3H] ORG-2058. Binding of steroids to receptors was a function of cellular uptake. Incubation periods of 50 min were sufficient to attain maximum intracellular incorporation. The binding of 17 beta-E2 and ORG-2058 to MCF-7 cells, a phenomenon which is saturable at low concentrations for the radioactive ligand, is a linear function of the number of cells assayed (Interval: 2.5 X 10(4) to 1.5 X 10(6) cells per well). Binding data and their Scatchard plot allowed for the calculation of affinity and capacity values. Thus, for ER, Kd = 2.0 +/- 0.5 X 10(-10) M and n = 3.76 +/- 0.91 Fmol/microgram DNA, and for PgR Kd = 2.0 +/- 0.2 X 10(-10) M and n = 14.02 +/- 2.30 Fmol/microgram DNA (Mean +/- SD). Binding specificity of 17 beta-Estradiol and ORG-2058 to MCF-7 cells was analysed by means of study on the inhibitory effect of increasing concentrations of unlabelled competitors: 17 beta-Estradiol, ORG-2058, Estrone, DES, R-5020, Cortisol, Androsterone and Testosterone. Only pharmacological doses of some of the mentioned molecules produce displacement of the hormonereceptor binding. This phenomenon appears to be related to the affinity of these chemical compounds for the receptor macromolecules to which estrogens and progesterone bind.

  2. Microarray-based determination of estrogen receptor, progesterone receptor, and HER2 receptor status in breast cancer

    NARCIS (Netherlands)

    P. Roepman; H.M. Horlings; O. Krijgsman; M. Kok; J.M. Bueno-de-Mesquita; R. Bender; S.C. Linn; A.M. Glas; M.J. van de Vijver

    2009-01-01

    Purpose: The level of estrogen receptor (ER), progesterone receptor (PR), and HER2 aids in the determination of prognosis and treatment of breast cancer. Immunohistochemistry is currently the predominant method for assessment, but differences in methods and interpretation can substantially affect th

  3. Plasma concentration of progesterone and 17ß-estradiol of black-rumped agouti (Dasyprocta prymnolopha during the estrous cycle

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    Diva Anelie Guimarães

    2011-03-01

    Full Text Available Plasma concentration of progesterone and 17β-estradiol of black-rumped agouti (Dasyprocta prymnolopha during the estrous cycle. The agouti is a game animal that have been raised in captivity for conservation and sustainability purposes. However, the management of wild animals in an intensive breeding system requires an assertive knowledge of its reproductive parameters, one of the most important features for production improvement. Besides, little information is available regarding changes in reproductive hormone profiles in agouti. The objective of this study was to evaluate the hormonal profile of progesterone and 17β-estradiol during the estrous cycle of the agouti (Dasyprocta prymnolopha. The hormones were analyzed by radioimmunoassay. Blood samples were collected without sedation twice a week. The concentrations of progesterone were as follows: proestrus 0.78+0.39ng/ml, estrus 2.83+2.34ng/ml, metestrus 1.49+1.24ng/ml, diestrus 3.71+1.48ng/ml. In the estrous phase, an increase in the progesterone level was observed during a period of 24h. The average 17 β-estradiol levels were as follows: proestrus 2 030.98+961.00pg/ml, estrus 1 910.56+650.54pg/ml, metestrus 1 724.83+767.28pg/ml, diestrus 1 939.94+725.29pg/ml. The current results suggest that the progesterone plasma concentration during the estrous cycle in the agouti has a similar increasing, stabilizing and decreasing pattern, as in domestic mammals. Agoutis have two phases of follicular development, as two periods of 17β-estradiol peaks were observed, the first one in the metestrus and the second during the proestrus. Spontaneous ovulation seems to occur after the progesterone peak, possibly indicating that this hormone is associated with the ovulatory process. A more detailed investigation is needed for better understanding of how progesterone influences ovulation. Studies on the involvement of progesterone in follicular rupture can be carried out, using steroid biosynthesis

  4. GnRH treatment at artificial insemination in beef cattle fails to increase plasma progesterone concentrations or pregnancy rates.

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    Perry, G A; Perry, B L

    2009-03-15

    Treatment with GnRH at the onset of standing estrus increased pregnancy percentages and circulating concentrations of progesterone in repeat breeder dairy cows. The objective of this study was to determine the effect of treatment with GnRH at AI on concentrations of progesterone and conception rates in beef cattle that exhibited estrus. Two hundred ninety-three heifers at four locations were synchronized with the Select Synch plus CIDR protocol (given GnRH and a CIDR was placed into the vagina, and 7 d later, given PGF(2alpha) and CIDR removed; n=253) or the 14-19 melengestrol acetate (MGA) protocol (MGA fed at 0.5mg/head/d for 14 d, with PGF(2alpha) 19 d after MGA withdrawal n=40) and AI was done after detection of estrus. At Location 1, blood samples were collected on Day 2, 4, 6, 10, 15, and 18 after AI (Day 0=AI). Two hundred and fifty postpartum cows at two locations were synchronized with the Select Synch plus CIDR protocol, and AI was performed after detection of estrus. At AI, cattle were alternately assigned to one of two treatments: (1) treatment with GnRH (100microg) at AI (n=127 heifers and n=108 cows); or (2) non-treated control (n=120 heifers and n=119 cows). Concentrations of progesterone tended to be greater in control heifers compared to GnRH-treated heifers on Days 6 (P=0.08), 10 (P=0.07), and 15 (P=0.11). Overall conception rates were 68% and 66% for GnRH treated and control, respectively, and were not different between treatments (P=0.72). In summary, treatment with GnRH at time of AI had no influence on conception rates in cattle that had exhibited estrus.

  5. Prediction of ovulation in women using a rapid progesterone radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Fleming, R.; Coults, J.R.T. (Glasgow Univ. (UK))

    1982-02-01

    A rapid (3-h) radioimmunoassay of plasma progesterone has been developed and used successfully to predict the time of ovulation in women undergoing artificial insemination. The results obtained using progesterone levels to date the stage of the cycle were analysed retrospectively by (1) estimation of the length of the ensuing luteal phases and comparison of these with luteal phase lengths of a control group (2) comparison of the dating using progesterone levels with retrospective determination of LH values and (3) by analysis of the dating in cycles in which conception occurred.

  6. Genetic determinants of plasma triglycerides

    Science.gov (United States)

    Johansen, Christopher T.; Kathiresan, Sekar; Hegele, Robert A.

    2011-01-01

    Plasma triglyceride (TG) concentration is reemerging as an important cardiovascular disease risk factor. More complete understanding of the genes and variants that modulate plasma TG should enable development of markers for risk prediction, diagnosis, prognosis, and response to therapies and might help specify new directions for therapeutic interventions. Recent genome-wide association studies (GWAS) have identified both known and novel loci associated with plasma TG concentration. However, genetic variation at these loci explains only ∼10% of overall TG variation within the population. As the GWAS approach may be reaching its limit for discovering genetic determinants of TG, alternative genetic strategies, such as rare variant sequencing studies and evaluation of animal models, may provide complementary information to flesh out knowledge of clinically and biologically important pathways in TG metabolism. Herein, we review genes recently implicated in TG metabolism and describe how some of these genes likely modulate plasma TG concentration. We also discuss lessons regarding plasma TG metabolism learned from various genomic and genetic experimental approaches. Treatment of patients with moderate to severe hypertriglyceridemia with existing therapies is often challenging; thus, gene products and pathways found in recent genetic research studies provide hope for development of more effective clinical strategies. PMID:21041806

  7. Plasma progesterone, metabolic hormones and beta-hydroxybutyrate in Holstein-Friesian cows after superovulation.

    Science.gov (United States)

    Bényei, Balázs; Komlósi, István; Pécsi, Anna; Kulcsár, Margit; Huzsvai, László; Barros, C W C; Huszenicza, Gyula

    2011-12-01

    Metabolic hormones [insulin, leptin, insulin-like growth factor-I (IGF-I), thyroxine (T4) and triiodothyronine (T3)], progesterone (P4) and beta-hydroxybutyrate (BHB) serum concentrations were evaluated and their effect on the superovulation results of donor cows was investigated in a semi-arid environment. Body weight, body condition score (BCS) and lactation stage were also included in the analysis. Twenty-three Holstein-Friesian cows were superovulated with 600 IU FSHp following the routine procedure and flushed on day 7 in a Multiple Ovulation and Embryo Transfer Centre in the semi-arid area of Brazil. The corpora lutea (CL) were counted and blood samples were collected for assays. All of the hormones investigated and BHB serum concentrations were within the physiological ranges. There was a positive correlation between hormones, except between BHB and all the others. The leptin level was influenced by feeding status, as indicated by the BCS. Insulin, T4, T3 and BHB levels were affected by milking status. Dry cows had higher levels of all hormones except BHB. An optimum level of leptin resulted in the highest number of CL, while the linear increase of P4, T4 and IGF significantly increased the number of CL.

  8. Plasma oxidative stress biomarkers and progesterone profiles in a dairy cow diagnosed with an ovarian follicular cyst.

    Science.gov (United States)

    Talukder, S; Ingenhoff, L; Kerrisk, K L; Celi, P

    2014-01-01

    This study was conducted to examine the oxidative stress biomarkers in a cow diagnosed with a follicular cyst in her left ovary. Progesterone (P4) and plasma oxidative stress status was measured in 13 Holstein cows after synchronization of oestrus with controlled internal drug release (CIDR) and prostaglandinF2α (PGF2α) protocol. The presence and size of ovarian structures were monitored by transrectal ultrasound at 4 hourly intervals. Of the 13 cows, 12 were monitored until ovulation was detected and recorded, whereas one cow failed to ovulate and developed a follicular cyst. Oxidative stress biomarkers; reactive oxygen metabolites (ROMs), biological antioxidant potential (BAP), oxidative stress index (OSI), glutathione (GSH), ceruloplasmin and advanced oxidation protein products (AOPP) were measured in the cystic cow and compared to those of the 12 ovulated cows and are referred to as higher or lower if they are outside the mean ± standard error of mean of those of ovulated cows. The cystic cow had lower ROMs and OSI between 36 and 84 h after PGF2α injection and at 9 h, from 36 to 60 h after PGF2α injection respectively. On the other hand, antioxidant (BAP and GSH) was higher in the cystic cow compared to her ovulated herd mates. The observed imbalance between oxidant and antioxidant might have disrupted the physiological events for ovulation to occur, leading to cystic ovarian disease.

  9. Development and validation of a liquid chromatographic method for the simultaneous determination of estradiol, estriol, estrone, and progesterone in pharmaceutical preparations.

    Science.gov (United States)

    Wilson, Phyllis

    2009-01-01

    Progesterone and estrogens are hormones produced in the human body that are essential for regulating many vital functions. The three major estrogens produced by women are estriol, estradiol, and estrone. Progesterone is a naturally occurring hormone in both men and women. Pharmaceuticals containing estrogens alone or estrogens in combination with progesterone are commonly used in therapy. Patients requiring unique combinations of the drugs rely on pharmacies to compound the ingredients. In order to assess the potency of drugs containing combinations of estrogens and progesterone, a method was developed to determine all four ingredients simultaneously. The liquid chromatographic method utilized a microBondapak C18 column with an isocratic mobile phase of acetonitrile-water (50 + 50, v/v) at a flow rate of 1.0 mL/min and temperature of 30 degrees C. Under these conditions, the order of elution was estriol, estradiol, and estrone, followed by progesterone. UV detection was at 205 nm to monitor elution of the estrogens, then switched to 270 nm to monitor progesterone. The method was applied to the analysis of pharmacy-compounded drugs containing combinations of the hormones. Validation studies demonstrated that the method is accurate and precise.

  10. [Sex hormones in young pigs. I. Concentrations of plasma progesterone and estrogens of sows and urinary estrogens after synchronization of estrus and ovulation].

    Science.gov (United States)

    Schneider, F; Bergfeld, J; Brauer, M; Brüssow, K P

    1980-01-01

    Treatment for synchronised heat and ovulation, using Suisynchron-Prämix, Bernburg, pregnant-mare serum gonadotrophin (PMSG), and human chorionic gonadotrophin (HCG), was followed with twelve gilts by daily measurement over 31 days of urine oestrogens, plasma progesterone as well as of plasma oestrogens during the oestric phases. Hormonal developments recorded during the oestrus and luteum phases were in agreement with the concepts and data so far known for the subject. An intermediate peak of oestrogen concentration was clearly recordable in the di-oestrus, that is between the tenth and 15th days of cycle. Its physiological importance will have to be further investigated.

  11. Effects of luteectomy in early pregnancy on the maintenance of gestation and plasma progesterone concentrations in the viviparous temperate lizard Barisia imbricata imbricata

    Directory of Open Access Journals (Sweden)

    Moreno-Fierros Leticia

    2010-02-01

    Full Text Available Abstract Background Several studies have shown that the corpus luteum is the principal source of progesterone during the gravidity period in reptiles; however, its participation in the maintenance of gestation in the viviparous squamata is in dispute. The effects of ovariectomy or luteectomy vary according to the species and the time at which the procedure is performed. In this paper, we describe the effects of luteectomy during early pregnancy on the maintenance of gestation and progesterone concentrations in the temperate Mexican viviparous lizard Barisia imbricata imbricata. Methods Twenty-four lizards were subjected to three different treatments: luteectomy, sham luteectomy or non-surgical treatment, and blood samples were obtained before and after surgical treatment at different stages of gestation to determine the effects of luteectomy on the maintenance of gestation and progesterone concentrations. Results Spontaneous abortion was not observed in any of the females. However, luteectomy provoked abnormal parturition and a significant reduction in the number of young born alive. Parturition was normal in untreated females as well as those submitted to sham luteectomy. The surgical treatment also caused a significant reduction in progesterone concentrations in luteectomised females during early and middle gestation. However, no significant differences in hormone concentrations were observed among the three groups during late gestation or immediately post-parturition. Conclusions Our observations indicate that the presence of the corpus luteum is not necesary for the maintenance of gestation, but that it does participate in parturition control. Moreover, the corpus luteum of the viviparous lizard B. i. imbricata produces progesterone, at least during the first half of pregnancy, and that an extra-ovarian source of progesterone must maintain gestation in the absence of luteal tissue.

  12. Desarrollo de un método inmunoenzimático para determinar progesterona Development of an immunoenzimatic method to determine progesterone

    Directory of Open Access Journals (Sweden)

    Roberto M. González Suárez

    2002-08-01

    Full Text Available Se desarrolló un método inmunoenzimático, competitivo, en fase líquida, con separación por anticuerpos acoplados a partículas magnetizables, para determinar progesterona en plasma; se basa en la competencia por los sitios de unión de un anticuerpo monoclonal, entre la hormona presente en la muestra y un conjugado hormona-enzima, en presencia de agentes bloqueadores de las proteínas de transporte de esteroides del plasma. El complejo antígeno-anticuerpo es atrapado por un segundo anticuerpo antiglobulina de ratón acoplado a partículas magnetizables que permite separarlo de los componentes no reaccionantes del sistema y desarrollar en su superficie la reacción de color con la que se cuantifica el analito. Este sistema no requiere extracción de los esteroides del plasma con solventes orgánicos, ni centrifugación y el producto final de la reacción puede ser leído en un colorímetro convencional. Todo lo anterior lo hace accesible al equipamiento de un laboratorio de bioquímica clínica convencional. Se estudiaron los parámetros de calibración y de calidad de ambos métodos, se compararon con 2 sistemas analíticos comerciales similares y con los resultados de la determinación de progesterona en 200 muestras y se comprobó que no existían diferencias estadísticas entre ellos. Se concluyó que este sistema cuenta con los requisitos necesarios para ser incorporado como método de rutina en los laboratorios del Sistema Nacional de Salud.An immunoenzimatic, competitive liquid stage method , with separation by antibodies coupled to magnetizable particles was developed to determine progesterone in plasma, It is based on the competence for the binding sites of a monoclonal antibody between the hormone present in the sample and a hormone-enzime conjugate in the presence of blocking agents of the proteins transporting steroids from plasma. The antigen-antibody complex is trapped by a second mouse antiglobulin antibody coupled to

  13. Patterns of plasma corticotropin-releasing hormone, progesterone, estradiol, and estriol change and the onset of human labor.

    Science.gov (United States)

    Smith, Roger; Smith, Julia I; Shen, Xiaobin; Engel, Patricia J; Bowman, Maria E; McGrath, Shaun A; Bisits, Andrew M; McElduff, Patrick; Giles, Warwick B; Smith, David W

    2009-06-01

    Clinical prediction of preterm delivery is largely ineffective, and the mechanism mediating progesterone (P) withdrawal and estrogen activation at the onset of human labor is unclear. Our objectives were to determine associations of rates of change of circulating maternal CRH in midpregnancy with preterm delivery, CRH with estriol (E3) concentrations in late pregnancy, and predelivery changes in the ratios of E3, estradiol (E2), and P. A cohort of 500 pregnant women was followed from first antenatal visits to delivery during the period 2000-2004 at John Hunter Hospital, New South Wales, Australia, a tertiary care obstetric hospital. Unselected subjects were recruited (including women with multiple gestations) and serial blood samples obtained. CRH daily percentage change in term and preterm singletons at 26 wk, ratios E3/E2, P/E3, and P/E2 and the association between E3 and CRH concentrations in the last month of pregnancy (with spontaneous labor onset) were assessed. CRH percentage daily change was significantly higher in preterm than term singletons at 26 wk (medians 3.09 and 2.73; P = 0.003). In late pregnancy, CRH and E3 concentrations were significantly positively associated (P = 0.003). E3/E2 increased, P/E3 decreased, and P/E2 was unchanged in the month before delivery (medians: E3/E2, 7.04 and 10.59, P onset of labor. Our evidence provides a rationale for the use of CRH in predicting preterm birth and informs approaches to delaying labor using P supplementation.

  14. [Plasma concentrations of folic acid, vitamin B12 and progesterone of cyclic bitches, bitches during pregnancy and induced abortion and bitches with pyometra].

    Science.gov (United States)

    Kalender, H; Beceriklisoy, H B; Kanca, H; Findik, M; Erünal-Maral, N; Handler, J; Aslan, S

    2006-09-01

    The aim of this study was to investigate the plasma concentrations of folic acid, vitamin B12 and progesterone at different stages of the sexual cycle and pregnancy, during induced abortion and in bitches with pyometra. Bitches (n = 97) were assigned to groups as follows: a) oestrous cycle (n = 42) b) pregnancy (n = 25) c) induction of abortion (n = 10 and d) pyometra (n = 20). Oestrous cycle stages were determined by vaginal inspection and cytology. Pregnancies were estimated by ultrasound (5.0 Mhz; linear transducer; Schimadzu) at days 15-25, 35-45 and 46-63 of pregnancy. Treatments for the induction of abortion were started between days 25 and 35 after mating (5 microg/kg cabergoline daily, Galastop; 5-10 microg/kg Alfaprostol every other day, Gabbrostim). Diagnosis of pyometra was confirmed by ultrasound and vaginoscopy. Folic acid and vitamin B12 concentrations did not differ among different stages of the oestrous cycle. The mean concentration of folic acid during early pregnancy (days 15-25) exceeded levels of later stages (days 46-63): 9.4 +/- 3.7 microg/ml and 4.7 +/- 1.8 microg/ml, respectively (p dogs ( r = 0.925; p pyometra significantly differed from that of bitches at different stages of the oestrous cycle (p pyometra (p < 0.05). The decrease of serum concentrations of folic acid during pregnancy and induced abortion show that fetal growth and abortion caused higher consumption of folic acid. Concerning bitches did not show any deficiency symptoms, which is why it can be concluded that this decrease is physiological.

  15. Steroid determination in fish plasma using capillary electrophoresis

    Science.gov (United States)

    Bykova, L.; Archer-Hartmann, S. A.; Holland, L.A.; Iwanowicz, L.R.; Blazer, V.S.

    2010-01-01

    A capillary separation method that incorporates pH-mediated stacking is employed for the simultaneous determination of circulating steroid hormones in plasma from Perca flavescens (yellow perch) collected from natural aquatic environments. The method can be applied to separate eight steroid standards: progesterone, 17α,20β-dihydroxypregn-4-en-3-one, 17α-hydroxyprogesterone, testosterone, estrone, 11-ketotestosterone, ethynyl estradiol, and 17β-estradiol. Based on screening of plasma, the performance of the analytical method was determined for 17α,20β-dihydroxypregn-4-en-3-one, testosterone, 11-ketotestosterone, and 17β-estradiol. The within-day reproducibility in migration time for these four steroids in aqueous samples was ≤2%. Steroid quantification was accomplished using a calibration curve obtained with external standards. Plasma samples from fish collected from the Choptank and Severn Rivers, Maryland, USA, stored for up to one year were extracted with ethyl acetate and then further processed with anion exchange and hydrophobic solid phase extraction cartridges. The recovery of testosterone and 17β-estradiol from yellow perch plasma was 84 and 85%, respectively. Endogenous levels of testosterone ranged from 0.9 to 44 ng/ml, and when detected 17α,20β-dihydroxypregn-4-en-3-one ranged from 5 to 34 ng/ml. The reported values for testosterone correlated well with the immunoassay technique. Endogenous concentrations of 17β-estradiol were ≤1.7 ng/ml. 11-Ketotestosterone was not quantified because of a suspected interferant. Higher levels of 17α,20β-dihydroxypregn-4-en-3-one were found in male and female fish in which 17β-estradiol was not detected. Monitoring multiple steroids can provide insight into hormonal fluctuations in fish.

  16. Radiometric assay for cytochrome P-450-catalyzed progesterone 16 alpha-hydroxylation and determination of an apparent isotope effect

    Energy Technology Data Exchange (ETDEWEB)

    Osawa, Y.; Coon, M.J.

    1987-08-01

    In the course of studies on the oxygenation of steroids by purified P-450 cytochromes, particularly rabbit liver microsomal cytochrome P-450 form 3b, a rapid and reliable radiometric assay has been devised for progesterone 16 alpha-hydroxylation. In view of the lack of a commercially available, suitably tritiated substrate, (1,2,6,7,16,17-3H)progesterone was treated with alkali to remove the label from potential hydroxylation sites other than the 16 alpha position. The resulting (1,7,16-3H)progesterone was added to a reconstituted enzyme system containing cytochrome P-450 form 3b, NADPH-cytochrome P-450 reductase, and NADPH, and the rate of 16 alpha-hydroxylation was measured by the formation of /sup 3/H/sub 2/O. This reaction was shown to be linear with respect to time and to the cytochrome P-450 concentration. An apparent tritium isotope effect of 2.1 was observed by comparison of the rates of formation of tritium oxide and 16 alpha-hydroxyprogesterone, and the magnitude of the isotope effect was confirmed by an isotope competition assay in which a mixture of (1,7,16-/sup 3/H)progesterone and (4-14C)progesterone was employed.

  17. Comparative efficacy of different estrus synchronization protocols on estrus induction response, fertility and plasma progesterone and biochemical profile in crossbred anestrus cows

    Directory of Open Access Journals (Sweden)

    A. J. Dhami

    2015-11-01

    Full Text Available Aim: To evaluate estrus induction response and fertility including plasma progesterone and biochemical profile following use of three standard hormonal protocols in anestrus crossbred cows. Materials and Methods: The study was carried out on 40 true anestrus and 10 normal cyclic cows. 10 anestrus cows each were treated with standard intravaginal controlled internal drug release (CIDR device, Ovsynch (GPG protocol, and Norgestomet ear implant with fixed-time artificial insemination (FTAI. 10 anestrus cows were kept as untreated control while 10 cows exhibiting the first estrus within 90 days postpartum without any treatment served as normal cyclic control. Blood samples were obtained from treated cows on day 0, 7, 9 (AI of treatment and day 21 post-AI, and from control groups on the day of AI and day 21 post-AI for estimation of plasma progesterone, protein, cholesterol, calcium, and inorganic phosphorus profile. Results: The use of CIDR, Ovsynch, and Norgestomet ear implant protocols resulted in 100% estrus induction with conception rates at induced estrus of 60%, 50%, and 50%, and the overall of three cycles as 80%, 80%, and 70%. In untreated anestrus control (n=10, only three cows exhibited spontaneous estrus within 90 days of follow-up and conceived giving the first service and overall conception rates of 66.66% and 30.00%, respectively. In normal cyclic control (n=10, the conception rates at first and overall of three cycles were 50% and 80%. The overall mean plasma progesterone (P4 concentrations in anestrus cows studied on day 0 (initiation, 7 (prostaglandin injection and/or removal of implant, 9 (FTAI of treatment and on day 21 post-AI revealed that the values on day 7 and 21 were significantly (p<0.01 higher than other two periods in all three groups. The concentrations were significantly (p<0.05 higher in conceived than non-conceived group on day 21 post-AI in CIDR (4.36±0.12 vs. 1.65±0.82 ng/ml and Ovsynch (4.85±0.62 vs. 1.59±0

  18. Gas chromatographic determination of xanthinol in plasma

    NARCIS (Netherlands)

    Roseboom, H.; Wiese, G.

    1979-01-01

    The determination of xanthinol in plasma is described. After extraction of the drug, together with the internal standard (papaverine hydrochloride), the extract is evaporated to dryness and the drug is derivatized with acetic anhydride for chromatography. The method is linear for 2–100μg ml-1 ; the

  19. Neither plasma progesterone concentrations nor exogenous eCG affects rates of ovulation or pregnancy in fixed-time artificial insemination (FTAI) protocols for puberal Nellore heifers.

    Science.gov (United States)

    Pegorer, M Figueira; Ereno, R L; Satrapa, R A; Pinheiro, V G; Trinca, L A; Barros, C M

    2011-01-01

    The objective was to evaluate the effects of plasma progesterone (P4) concentrations and exogenous eCG on ovulation and pregnancy rates of pubertal Nellore heifers in fixed-time artificial insemination (FTAI) protocols. In Experiment 1 (Exp. 1), on Day 0 (7 d after ovulation), heifers (n = 15) were given 2 mg of estradiol benzoate (EB) im and randomly allocated to receive: an intravaginal progesterone-releasing device containing 0.558 g of P4 (group 0.5G, n = 4); an intravaginal device containing 1 g of P4 (group 1G, n = 4); 0.558 g of P4 and PGF(2α) (PGF; 150 μg d-cloprostenol, group 0.5G/PGF, n = 4); or 1 g of P4 and PGF (group 1G/PGF, n = 3). On Day 8, PGF was given to all heifers and intravaginal devices removed; 24 h later (Day 9), all heifers were given 1 mg EB im. In Exp. 2, pubertal Nellore heifers (n = 292) were treated as in Exp. 1, with FTAI on Day 10 (30 to 36 h after EB). In Exp. 3, pubertal heifers (n = 459) received the treatments described for groups 0.5G/PGF and 1G/PGF and were also given 300 IU of eCG im (groups 0.5G/PGF/eCG and 1G/PGF/eCG) at device removal (Day 8). In Exp. 1, plasma P4 concentrations were significantly higher in heifers that received 1.0 vs 0.588 g P4, and were significantly lower in heifers that received PGF on Day 0. In Exp. 2 and 3, there were no significant differences among groups in rates of ovulation (65-77%) or pregnancy (Exp. 2: 26-33%; Exp. 3: 39-43%). In Exp. 3, diameter of the dominant ovarian follicle on Day 9 was larger in heifers given 0.558 g vs 1.0 g P4 (10.3 ± 0.2 vs 9.3 ± 0.2 mm; P decreased plasma P4 from Days 1 to 8 and increased diameter of the dominant follicle on Day 9. However, neither of these nor 300 IU of eCG on Day 8 significantly increased rates of ovulation or pregnancy.

  20. The influence of nutrition on the insulin-like growth factor system and the concentrations of growth hormone, glucose, insulin, gonadotropins and progesterone in ovarian follicular fluid and plasma from adult female horses (Equus caballus).

    Science.gov (United States)

    Salazar-Ortiz, Juan; Monget, Philippe; Guillaume, Daniel

    2014-07-31

    Feed intake affects the GH-IGF system and may be a key factor in determining the ovarian follicular growth rate. In fat mares, the plasma IGF-1 concentration is high with low GH and a quick follicular growth rate, in contrast to values observed in thin mares. Nothing is known regarding the long-term effects of differential feed intake on the IGF system. The objective of this experiment was to quantify IGFs, IGFBPs, GH, glucose, insulin, gonadotropin and progesterone (P4) in blood and in preovulatory follicular fluid (FF) in relation to feeding levels in mares. Three years prior to the experiment, Welsh Pony mares were assigned to a restricted diet group (R, n = 10) or a well-fed group (WF, n = 9). All mares were in good health and exhibited differences in body weight and subcutaneous fat thickness. Follicular development was scanned daily and plasma was also collected daily. Preovulatory FF was collected by ultrasound-guided follicular aspiration. Hormone levels were assayed in FF and plasma with a validated RIA. According to scans, the total number of follicles in group R was 53% lower than group WF. Insulin and IGF-1 concentrations were higher in WF than in R mares. GH and IGF-2 concentrations were lower in plasma from WF mares than from R mares, but the difference was not significant in FF. The IGFBP-2/IGFBP-3 ratio in FF was not affected by feeding but was dramatically increased in R mare plasma. No difference in gonadotropin concentration was found with the exception of FSH, which was higher in the plasma of R mares. On the day of puncture, P4 concentrations were not affected by feeding but were higher in preovulatory FF than in plasma. The bioavailability of IGF-1 or IGF-2, represented by the IGFBP2/IGFBP3 ratio, is modified by feed intake in plasma but not in FF. These differences partially explain the variability in follicular growth observed between well-fed mares and mares on restricted diets.

  1. Determination and distribution of human plasma selenoproteins

    Energy Technology Data Exchange (ETDEWEB)

    Plecko, T.; Nordmann, S.; Ruekgauer, M.; Kruse-Jarres, J.D. [Institute for Clinical Chemistry and Laboratory Medicine, Stuttgart (Germany)

    1999-03-01

    Major portions of plasma-selenium are incorporated in the proteins glutathione peroxidase (GSH-Px), selenoprotein P (Sel P) and albumin. A chromatographic method, adapted from a procedure by Harrison et al. [6], uses heparin- and blue-sepharose to separate the three protein fractions. The determination of selenium was carried out by electrothermal atomic absorption spectroscopy (ETAAS) using the Zeeman effect. The selenium distribution of 17 healthy subjects was 68 {+-} 7% of the total plasma selenium associated to Sel P, 25 {+-} 4% associated to p-GSH-Px and 7{+-}4% associated to albumin. The recovery of selenium was 99 {+-} 4%. For precision measurements a plasma pool has been separated seven times. The selectivity of this method was monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and GSH-Px activity measurements. A fast method, adapted for clinical applications, is described which allows to determine the human plasma selenium distribution in about an hour. (orig.) With 2 tabs., 8 refs.

  2. Correlation of brain levels of progesterone and dehydroepiandrosterone with neurological recovery after traumatic brain injury in female mice.

    Science.gov (United States)

    Lopez-Rodriguez, Ana Belen; Acaz-Fonseca, Estefania; Giatti, Silvia; Caruso, Donatella; Viveros, Maria-Paz; Melcangi, Roberto C; Garcia-Segura, Luis M

    2015-06-01

    Traumatic brain injury (TBI) is an important cause of disability in humans. Neuroactive steroids, such as progesterone and dehydroepiandrosterone (DHEA), are neuroprotective in TBI models. However in order to design potential neuroprotective strategies based on neuroactive steroids it is important to determine whether its brain levels are altered by TBI. In this study we have used a weight-drop model of TBI in young adult female mice to determine the levels of neuroactive steroids in the brain and plasma at 24h, 72 h and 2 weeks after injury. We have also analyzed whether the levels of neuroactive steroids after TBI correlated with the neurological score of the animals. TBI caused neurological deficit detectable at 24 and 72 h, which recovered by 2 weeks after injury. Brain levels of progesterone, tetrahydroprogesterone (THP), isopregnanolone and 17β-estradiol were decreased 24h, 72 h and 2 weeks after TBI. DHEA and brain testosterone levels presented a transient decrease at 24h after lesion. Brain levels of progesterone and DHEA showed a positive correlation with neurological recovery. Plasma analyses showed that progesterone was decreased 72 h after lesion but, in contrast with brain progesterone, its levels did not correlate with neurological deficit. These findings indicate that TBI alters the levels of neuroactive steroids in the brain with independence of its plasma levels and suggest that the pharmacological increase in the brain of the levels of progesterone and DHEA may result in the improvement of neurological recovery after TBI.

  3. Effect of nutrition on plasma progesterone levels, metabolic parameters and small follicles development in unstimulated goats reared under constant photoperiod regimen.

    Science.gov (United States)

    Rondina, D; Freitas, V J F; Spinaci, M; Galeati, G

    2005-12-01

    Sixteen local adult goats were submitted for 9 weeks to 2.09 (high group) and 0.54 (low group) x dietary maintenance respectively. During the experimental period, goats were weighed, oestrus was detected and plasma insulin, urea, non-esterified fatty acids and progesterone concentrations were assessed. At the end of the experiment, ovarian small follicles population was studied by histological analysis. Final weight loss in low group was 18.37 +/- 2.02%, whereas weight gain of high group was 13.84 +/- 2.70%. Insulin and urea were lower in low group, while non-esterified fatty acids were significantly higher. A lower number of fasted goats was in oestrus or ovulated and an extended length of oestrus (p goats a significant qualitative alteration of follicle classes involved in the initiation process of primordial pool was found. In this phase, granulosa thickness and oocyte size were the most affected (p 0.05). Collectively, these results indicate that opposite dietary intakes for a medium period induce a composite reproductive response in goats and can regulate the early onset of follicle growth.

  4. HPLC determination of tolperisone in human plasma.

    Science.gov (United States)

    Bae, Jung-Woo; Park, Young-Seo; Sohn, Uy-Dong; Myung, Chang-Sun; Ryu, Byung-Kwon; Jang, Choon-Gon; Lee, Seok-Yong

    2006-04-01

    A simple high performance liquid chromatographic (HPLC) method was developed for the determination of tolperisone in human plasma. Tolperisone and internal standard (chlorphenesin) were isolated from 1 mL of plasma using 8 mL of dichlormethane. The organic phase was collected and evaporated under nitrogen gas. The residue was then reconstituted with 300 mL aliquot of mobile phase and a 100 mL aliquot was injected onto the C18 reverse-phased column. The mobile phase, 45% methanol containing 1% glacial acetic acid and 0.05% 1-hexanesulfonic acid was run at a flow rate of 1 mL/min. The column effluent was monitored using UV detector at 260 nm. The retention times for tolperisone and the internal standard were approximately 7.1 and 8.4 min, respectively. The standard curve was linear with minimal intra-day and inter-day variability. The quantification limit of tolperisone in human plasma was 10 ng/ mL. The proposed method has been applied to the determination of pharmacokinetic profile of tolperisone in Koreans. The Tmax of tolperisone in Koreans (0.94 +/- 0.42 h) was not significantly differ from that reported in Europeans (0.5-1 h), but the mean half-life in Koreans (1.14 +/- 0.27 h) was shorter than that in Europeans (2.56 +/- 0.2 h). The proposed HPLC method is simple, accurate, reproducible and suitable for pharmacokinetic study of tolperisone.

  5. Abnormal regulation for progesterone production in placenta with prenatal cocaine exposure in rats.

    Science.gov (United States)

    Wu, L; Yan, J; Qu, S C; Feng, Y Q; Jiang, X L

    2012-12-01

    Cocaine abuse in pregnant women is currently a significant public hygiene problem and is tightly associated with elevated risk for preterm delivery. Placental steroidogenesis especially progesterone production was essential for success and maintenance of pregnancy in humans and rodents. In the present study, we determined the impact of prenatal cocaine exposure on pathways of placental progesterone synthesis in rats. Pregnant rats were treated cocaine twice daily (15 mg/kg/day) during the third trimester, and the maternal and fetal plasma progesterone and pregnenolone concentrations were detected. We also examined both the protein and mRNA expression of some key enzymes and regulators for progesterone production in placenta. Results showed that, after maternal cocaine use during pregnancy, progesterone and pregnenolone concentrations in both maternal and fetal rats were significantly decreased. Although prenatal cocaine exposure had no effects on placental 3β-hydroxysteroid dehydrogenase type 1 (3βHSD1) expression, protein and mRNA expression of the cholesterol side-chain cleavage enzyme (P450scc/CYP11a) in placenta was significantly inhibited. Moreover, protein and mRNA expressions of MLN64 that regulating cholesterol transport and activating protein 2γ (AP2γ/Tfap2c) that controlling P450scc/CYP11a gene expression in placenta were both decreased following maternal cocaine use in pregnancy. Collectively, this study suggested that prenatal cocaine exposure could insult the placental progesterone production in rats possibly associated with the high risk for preterm delivery.

  6. Sexual Absorption of Vaginal Progesterone: A Randomized Control Trial

    Directory of Open Access Journals (Sweden)

    Kathryn S. Merriam

    2015-01-01

    Full Text Available Objective. To determine if sexual intercourse reduces absorption of vaginal progesterone gel in women and to determine if progesterone is absorbed by the male during intercourse. Study Design. Prospective, randomized, cross over, controlled study of 20 reproductive-aged women and their male sexual partners randomized to receive vaginal progesterone gel (Crinone 8% gel, Actavis Inc., USA or placebo cream. Serum progesterone for both male and female partners were measured 10 hours after intercourse. One week later, subjects were crossed over to receive the opposite formulation. In the third week, women used progesterone gel at night and abstained from intercourse. Results. Serum progesterone was significantly reduced with vaginal progesterone gel + intercourse compared with vaginal progesterone gel + abstinence (P=0.0075. Men absorbed significant progesterone during intercourse with a female partner using vaginal progesterone gel compared to placebo (P=0.0008. Conclusion(s. Vaginal progesterone gel is reduced in women after intercourse which may decrease drug efficacy during luteal phase support. Because men absorb low levels of progesterone during intercourse, exposure could cause adverse effects such as decreased libido. This study is registered under Clinical Trial number NCT01959464.

  7. Patterns of Plasma Corticotropin-Releasing Hormone, Progesterone, Estradiol, and Estriol Change and the Onset of Human Labor

    National Research Council Canada - National Science Library

    Smith, Roger; Smith, Julia I; Shen, Xiaobin; Engel, Patricia J; Bowman, Maria E; McGrath, Shaun A; Bisits, Andrew M; McElduff, Patrick; Giles, Warwick B; Smith, David W

    2009-01-01

    ...) withdrawal and estrogen activation at the onset of human labor is unclear. Objectives: Our objectives were to determine associations of rates of change of circulating maternal CRH in midpregnancy with preterm delivery, CRH with estriol (E3...

  8. Effect of progesterone concentration and duration of proestrus on fertility in beef cattle after fixed-time artificial insemination.

    Science.gov (United States)

    Dadarwal, D; Mapletoft, R J; Adams, G P; Pfeifer, L F M; Creelman, C; Singh, J

    2013-03-15

    The objective was to determine the effect of plasma progesterone concentration and the duration of proestrus during growth of the ovulatory follicle on fertility in beef cattle. Heifers (N = 61) and postpartum cows (N = 79) were assigned randomly to four groups in a two-by-two design involving luteal-phase versus subluteal-phase plasma progesterone concentrations and normal versus short proestrus. To synchronize follicular wave emergence, estradiol-17β was given im during the midluteal phase (Day 0) and concurrently, a once-used controlled intravaginal progesterone-releasing device was placed intravaginally. In the subluteal-phase progesterone groups, a luteolytic dose of PGF(2α) was given on Day 0 and again 12 hours later. In the luteal-phase progesterone groups, PGF(2α) was not given (so as to retain a functional CL). The controlled intravaginal progesterone-releasing device was removed and PGF(2α) was given on Days 7 or 8 in the normal- and short-proestrus groups, respectively. Cattle were given lutropin im 12 or 36 hours later in the short- and normal-proestrus groups, respectively, with AI at 12 hours after lutropin treatment. Transrectal ultrasonography was used to monitor ovarian response during treatments and to diagnose pregnancy 60 days after AI. Cattle (heifers and cows combined) in the subluteal-phase progesterone groups and normal proestrus groups had a larger follicle at the time of AI, and a larger CL that secreted more progesterone 9 days after AI than cattle with luteal-phase progesterone concentrations or those with short proestrus (P < 0.03). There was a higher incidence of ovulation (P < 0.01) the day after AI in heifers (55/61; 90%) than in cows (44/79; 56%). Pregnancy rates ranged from 11% to 54%, and were higher in cattle (heifers and cows combined) in the subluteal-phase progesterone groups and normal proestrus groups than in the luteal-phase progesterone or short proestrus groups, respectively, (P < 0.02). In conclusion, a short

  9. Progesterone receptor expression declines in the guinea pig uterus during functional progesterone withdrawal and in response to prostaglandins.

    Directory of Open Access Journals (Sweden)

    Toni N Welsh

    Full Text Available Progesterone withdrawal is essential for parturition, but the mechanism of this pivotal hormonal change is unclear in women and other mammals that give birth without a pre-labor drop in maternal progesterone levels. One possibility suggested by uterine tissue analyses and cell culture models is that progesterone receptor levels change at term decreasing the progesterone responsiveness of the myometrium, which causes progesterone withdrawal at the functional level and results in estrogen dominance enhancing uterine contractility. In this investigation we have explored whether receptor mediated functional progesterone withdrawal occurs during late pregnancy and labor in vivo. We have also determined whether prostaglandins that induce labor cause functional progesterone withdrawal by altering myometrial progesterone receptor expression. Pregnant guinea pigs were used, since this animal loses progesterone responsiveness at term and gives birth in the presence of high maternal progesterone level similarly to primates. We found that progesterone receptor mRNA and protein A and B expression decreased in the guinea pig uterus during the last third of gestation and in labor. Prostaglandin administration reduced while prostaglandin synthesis inhibitor treatment increased progesterone receptor A protein abundance. Estrogen receptor-1 protein levels remained unchanged during late gestation, in labor and after prostaglandin or prostaglandin synthesis inhibitor administration. Steroid receptor levels were higher in the non-pregnant than in the pregnant uterine horns. We conclude that the decreasing expression of both progesterone receptors A and B is a physiological mechanism of functional progesterone withdrawal in the guinea pig during late pregnancy and in labor. Further, prostaglandins administered exogenously or produced endogenously stimulate labor in part by suppressing uterine progesterone receptor A expression, which may cause functional progesterone

  10. Immunohistochemical determination of estrogen and progesterone receptors in breast cancer: relationship with clinicopathologic factors in 302 patients in Ivory Coast.

    Science.gov (United States)

    Effi, Ahoua Benjamin; Aman, Nguiessan Alphonse; Koui, Baumaney Sylvanus; Koffi, Kouadio Donatien; Traoré, Zie Cheick; Kouyate, Mohamed

    2017-02-07

    Breast cancer is a heterogeneous and a hormone-dependent disease. The detection of the estrogen receptor (ER) and progesterone receptor (PgR) is crucial for prognostic evaluation and treatment choice of breast cancer for clinical practice. The purpose of this study was to evaluate the expression of the hormonal receptors, their distribution, and their correlation with clinicopathologic prognostic parameters for the improvement of the patients' treatment in Ivory Coast. The 20-month prospective study included 302 patients who were diagnosed with primary invasive breast carcinomas at the Central Laboratory in Abidjan. The paraffin-embedded blocks of these patients were examined by immunohistochemistry to assess the ER and PgR status. The one-way analysis of variance and Chi-Square Test were used to analyze the data. The mean age of patients at diagnosis was 48 ± 11 years. The majority of the women were premenopausal in 180 cases (59.9%). The predominant histologic type was invasive ductal carcinoma not otherwise specified (IDC NOS) in 247 cases (82%). Tumor grade 2 was more frequent in 166 cases (55%). Among 302 patients, 169 (56%) and 154 (49%) expressed ER and PgR respectively. The ER+PgR+ group with 131 cases (43%) was predominant, followed by 116 cases (38%) of ER-PgR-. The expression of ER and PgR was correlated with the age of the patients (p = 0.026) and the tumor grade (p = 0.0004). However, there was not statistically significant correlation between ER/PgR and the menopausal status of patients (p = 0.149), nor between ER/PgR and the histologic type (p = 0.523). The ER+PgR+ and ER-PgR- are the most common subgroups in women with breast cancer in Ivory Coast. The hormonal receptor status is associated with the age and the histologic grade in breast cancer patients. The systematic use of hormonal treatment should be reevaluated. A further study should be done to investigate the reasons of high rate of ER-PgR- in breast cancer patients in

  11. Superovulation of cows with PMSG: Variation in plasma concentrations of progesterone, oestradiol, LH, cortisol, prolactin and PMSG and in number of preovulatory follicles

    NARCIS (Netherlands)

    Bevers, M.M.; Dieleman, S.J.

    1987-01-01

    Forty-two heifers, exhibiting normal oestrous cycles, were treated with 2500 I.U. PMSG and 2 ml prostaglandin (PG) at day 10 and 12 of the oestrous cycle, respectively. In ten heifers progesterone, oestradiol, LH, prolactin, cortisol and PMSG levels were estimated until 10 days after the initiation

  12. Superovulation of cows with PMSG: Variation in plasma concentrations of progesterone, oestradiol, LH, cortisol, prolactin and PMSG and in number of preovulatory follicles

    NARCIS (Netherlands)

    Bevers, M.M.; Dieleman, S.J.

    Forty-two heifers, exhibiting normal oestrous cycles, were treated with 2500 I.U. PMSG and 2 ml prostaglandin (PG) at day 10 and 12 of the oestrous cycle, respectively. In ten heifers progesterone, oestradiol, LH, prolactin, cortisol and PMSG levels were estimated until 10 days after the initiation

  13. Determination of benzocaine in rainbow trout plasma

    Science.gov (United States)

    Bernardy, Jeffery A.; Coleman, K.S.; Stehly, G.R.; Gingerich, William H.

    1996-01-01

    A liquid chromatographic method is described for analysis of benzocaine (BZ), a proposed fish anesthetic, in rainbow trout plasma, Mean recoveries of BZ from plasma samples fortified at 44-10 100 ng/mL were 96-100%. The method detection limit is 10 ng/mL, and the limit of quantitation is 37 ng/mL. Acetylation of BZ occurs in whole blood after storage at room temperature (i.e., 21 degrees C) for 10 min. However, no acetylation of BZ was detected in plasma samples held at room temperature for 4 h, Mean method precision for plasma samples with incurred BZ residue is similar to that for fortified samples in the same concentration range (relative standard deviations of 0.9 and 1.2%, respectively).

  14. Autoimmune Progesterone Anaphylaxis

    Directory of Open Access Journals (Sweden)

    Mohammad Hassan Bemanian

    2007-06-01

    Full Text Available Progesterone induced dermatitis is a rare disorder. It typically occurs in females due to anautoimmune phenomenon to endogenous progesterone production, but can also be caused byexogenous intake of a synthetic progestin. Here in, we present a case of autoimmune progesterone anaphylaxis (AIPA observed in an adolescent female.The patient is an 18-year-old Caucasian female with no significant past medical history and noprior exogenous hormone use, who presented to her primary care physician complaining of cyclic skin eruptions with dyspnea, cough and respiratory distress. She noted that her symptoms occurred monthly, just prior to her menses. An intradermal skin test using 0.1 cml of progesterone was performed. The patient developed a 15mm wheal after 15 minutes, confirming the diagnosis of AIPA.The patient was started on a continuous regimen of an oral conjugated estrogen (0.625mg. The skin eruptions and respiratory symptoms have not returned since the initiation of this therapy.Autoimmune progesterone dermatitis manifests via the occurrence of cyclic skin eruptions.Women with the disorder commonly present with dermatologic lesions in the luteal phase of themenstrual cycle, if there are any other organ involvement in addition to skin (e.g. lung, GI thereaction should be called as autoimmune progesterone anaphylaxis. Diagnosis of AIPA is confirmed by performing a skin allergen test using progesterone.

  15. Give progesterone a chance

    Institute of Scientific and Technical Information of China (English)

    Florencia Labombarda; Daniel Garcia-Ovejero

    2014-01-01

    There is currently no standard pharmacological treatment for spinal cord injury. Here, we suggest that progesterone, a steroid hormone, may be a promising therapeutical candidate as it is already for traumatic brain injury, where it has reached phase II clinical trials. We rely on previous works showing anti-inflammatory, neuroprotective and promyelinating roles for progesterone after spinal cord injury and in our recent paper, in which we demonstrate that progesterone dimin-ishes lesion, preserves white matter integrity and improves locomotor recovery in a clinically relevant model of spinal cord lesion.

  16. Perfil hormonal de Progesterona durante o ciclo Estral em novilhas Nelore confinadas com Diferentes Ondas de Crescimento Folicular Plasma Progesterone Level during the Estrous Cycle in Nelore Heifers Confined with Two, Three and Four Waves of Follicular Growth

    Directory of Open Access Journals (Sweden)

    Luciene Lomas Santiago

    2001-12-01

    Full Text Available Efetuaram-se coletas diárias de sangue, de 16 novilhas Nelore confinadas, para análise de progesterona plasmática pelo método de radioimunoensaio (RIA. Os dias analisados para progesterona foram o dia zero (estro e a cada três dias até o dia -1 e o dia zero. Os animais foram divididos em dois grupos: 1 com ciclo estral de 21 dias aproximadamente (novilhas que apresentaram duas e três ondas de crescimento folicular e 2 com ciclo estral superior a 25 dias (novilhas com quatro ondas de crescimento folicular. As concentrações médias de progesterona plasmática dos animais durante o ciclo estral diferiram entre os dois grupos, sendo superiores (4,27 ng/mL para os ciclos de maior duração. A concentração média de progesterona no ciclo de aproximadamente 21 dias foi de 2,54 ng/mL. Os resultados sugerem que as novilhas que apresentam maior duração do ciclo estral necessitam de tempo adicional para que seus folículos cheguem ao estádio pré-ovulatório, havendo, dessa maneira, prolongamento e aumento da secreção de progesterona.Blood were collected daily from 16 Nelore heifers confined, for radioimmunoassay (RIA.analyses of progesterone The plasma progesterone assay were at day zero (estrus and at each three days until the -1 and the day zero.again The animals were divided in two groups: 1 with regular estrous cycle of 21 days (heifers with two and three follicular growth waves and 2 with prolonged estrous cycle, greater than 25 days (heifers with four follicular growth waves. The mean plasma progesterone level from the animals during the estrous cycle differed between the two groups, being greater (4,27 ng/mL for the extended cycles.(above 25 days; 4,27 ng/mL than for the regular estrous cycle (21 days; 2,54 ng/mL. Results suggest that those heifers which showed an extended estrous cycles, needs an additional time for the follicles to each the pre-ovulatory stadium, resulting in prolonged and increased progesterone secretion.

  17. Progesterone, BDNF and Neuroprotection

    OpenAIRE

    Singh, Meharvan; Su,Chang

    2012-01-01

    While the effects of progesterone in the central nervous system (CNS), like those of estrogen, have generally been considered within the context of reproductive function, growing evidence supports its importance in regulating non-reproductive functions including cognition and affect. In addition, progesterone has well-described protective effects against numerous insults in a variety of cell models, animal models and in humans. While ongoing research in several laboratories continues to shed ...

  18. Long-term exposure to electromagnetic radiation from mobile phones and Wi-Fi devices decreases plasma prolactin, progesterone, and estrogen levels but increases uterine oxidative stress in pregnant rats and their offspring.

    Science.gov (United States)

    Yüksel, Murat; Nazıroğlu, Mustafa; Özkaya, Mehmet Okan

    2016-05-01

    We investigated the effects of mobile phone (900 and 1800 MHz)- and Wi-Fi (2450 MHz)-induced electromagnetic radiation (EMR) exposure on uterine oxidative stress and plasma hormone levels in pregnant rats and their offspring. Thirty-two rats and their forty newborn offspring were divided into the following four groups according to the type of EMR exposure they were subjected to: the control, 900, 1800, and 2450 MHz groups. Each experimental group was exposed to EMR for 60 min/day during the pregnancy and growth periods. The pregnant rats were allowed to stand for four generations (total 52 weeks) before, plasma and uterine samples were obtained. During the 4th, 5th, and 6th weeks of the experiment, plasma and uterine samples were also obtained from the developing rats. Although uterine lipid peroxidation increased in the EMR groups, uterine glutathione peroxidase activity (4th and 5th weeks) and plasma prolactin levels (6th week) in developing rats decreased in these groups. In the maternal rats, the plasma prolactin, estrogen, and progesterone levels decreased in the EMR groups, while the plasma total oxidant status, and body temperatures increased. There were no changes in the levels of reduced glutathione, total antioxidants, or vitamins A, C, and E in the uterine and plasma samples of maternal rats. In conclusion, although EMR exposure decreased the prolactin, estrogen, and progesterone levels in the plasma of maternal rats and their offspring, EMR-induced oxidative stress in the uteri of maternal rats increased during the development of offspring. Mobile phone- and Wi-Fi-induced EMR may be one cause of increased oxidative uterine injury in growing rats and decreased hormone levels in maternal rats. TRPV1 cation channels are the possible molecular pathways responsible for changes in the hormone, oxidative stress, and body temperature levels in the uterus of maternal rats following a year-long exposure to electromagnetic radiation exposure from mobile phones and

  19. Progesterone Receptor Subcellular Localization and Gene Expression Profile in Human Astrocytoma Cells Are Modified by Progesterone

    Directory of Open Access Journals (Sweden)

    Aliesha González-Arenas

    2014-11-01

    Full Text Available Intracellular progesterone receptor (PR has been identified in human astrocytomas, the most common and aggressive primary brain tumors in humans. It has been reported that PR cell distribution affects their transcriptional activity and turnover. In this work we studied by immunofluorescence the effects of estradiol and progesterone on the subcellular localization of PR in a grade III human astrocytoma derived cell line (U373. We observed that total PR was mainly distributed in the cytoplasm without hormonal treatment. Estradiol (10 nM increased PR presence in the cytoplasm of U373 cells, whereas progesterone (10 nM and RU486 (PR antagonist, 1 μM blocked this effect. To investigate the role of PR activity in the regulation of gene expression pattern of U373 cells, we evaluated by microarray analysis the profile of genes regulated by progesterone, RU486, or both steroids. We found different genes regulated by steroid treatments that encode for proteins involved in metabolism, transport, cell cycle, proliferation, metastasis, apoptosis, processing of nucleic acids and proteins, adhesion, pathogenesis, immune response, cytoskeleton, and membrane receptors. We determined that 30 genes were regulated by progesterone, 41 genes by RU486 alone, and 13 genes by the cotreatment of progesterone+RU486, suggesting that there are many genes regulated by intracellular PR or through other signaling pathways modulated by progesterone. All these data suggest that PR distribution and activity should modify astrocytomas growth.

  20. Dynamics of Progesterone, TNF-α, and a Metabolite of PGF2α in Blood Plasma of Beef Cows following Embryo Transfer

    Directory of Open Access Journals (Sweden)

    M. C. Mason

    2014-01-01

    Full Text Available Lactating beef cows previously synchronized for estrus (d 0 were assigned to four treatments to assess their effectiveness in increasing blood progesterone (P4 and its effects on tumor necrosis factor-α (TNF-α and prostaglandin F2α (PGF2α after the transfer of embryos. At the time of transfer (d 7, cows received no treatment (control; n=16, a controlled internal drug releasing device (CIDR; n=16, human chorionic gonadotropin (hCG; n=15, or gonadotropin releasing hormone (GnRH; n=15. Blood samples were taken on d 7, 14, and 21 for analysis of P4 and tumor necrosis factor-α (TNF-α. Blood was collected (every 15 min for 2 h in half the animals in each treatment group on d 14 and the remaining half on d 21 for analysis of prostaglandin F2α metabolite (PGFM. Retention rates were 56.2, 62.5, 46.7, and 13.3% for cows in the control, CIDR, hCG, and GnRH groups, respectively. Progesterone was greater (P≤0.05 in cows receiving hCG compared to others on d 14. Progesterone in all treatment groups increased from d 7 to d 14 and declined (P≤0.05 from d 14 to d 21. Contrary to pregnant cows, P4 and TNF-α declined from d 7 to d 21 in nonpregnant cows (P≤0.05. Although PGFM increased by d 21, there was no difference between pregnant and nonpregnant cows.

  1. Use of a five-day progesterone-based timed AI protocol to determine if flunixin meglumine improves pregnancy per timed AI in dairy heifers.

    Science.gov (United States)

    Rabaglino, M B; Risco, C A; Thatcher, M-J; Lima, F; Santos, J E P; Thatcher, W W

    2010-06-01

    Two experiments were conducted to test the hypothesis that the 5 d Co-Synch + CIDR (Controlled Internal Drug Release insert containing progesterone) protocol could be applied as an efficient timed AI (TAI) protocol in dairy heifers, and that treatment with flunixin meglumine (FM) during the period of CL maintenance would increase pregnancy per TAI (P/TAI) and late survival of embryos. Objectives were: 1) in Experiment 1, to compare P/TAI with the 5 d Co-Synch+CIDR protocol to a PGF(2alpha)/GnRH protocol; and 2) in Experiment 2, to determine if FM administered 15.5 and 16 d after first TAI would increase P/TAI, using the 5 d Co-Synch+CIDR protocol with a new or previously used (5 d) CIDR insert. In Experiment 1, 248 heifers were assigned randomly to either the PGF(2alpha)/GnRH protocol (n=120) or the 5 d Co-Synch+CIDR protocol (n=128). Pregnancy per TAI did not differ between the 5 d Co-Synch+CIDR protocol (53.1%) and the PGF(2alpha)/GnRH protocol (45.8%; P=0.22). In Experiment 2, 325 heifers synchronized with the 5 d Co-Synch+CIDR protocol were assigned randomly to receive two injections of FM (FM group; n=158) at 15.5 and 16 d after TAI, or to remain as untreated controls (n=165). Pregnancy per TAI in Experiment 2 was 59.4 and 59.5% at 45 d for control and FM groups, respectively, with no differences between groups (P=0.83). The 5 d Co-Synch+CIDR protocol resulted in an acceptable P/TAI in dairy heifers. However, FM did not improve P/TAI in dairy heifers. 2010 Elsevier Inc. All rights reserved.

  2. Does the conceptus of the viviparous lizard Barisia imbricata imbricata participates in the regulation of progesterone production and the control of luteolysis?

    Science.gov (United States)

    Martínez-Torres, Martín; Salcedo-Álvarez, Martha; Alvarez-Rodríguez, Carmen; Cárdenas-León, Mario; Luis, Juana; Moreno-Fierros, Leticia

    2014-08-01

    It is generally accepted that progesterone is necessary to maintain gestation; however, the mechanisms that control the production of this steroid remain unknown. The corpus luteum has been assigned a central role in the maintenance of gestation based on its capacity to produce progesterone. A pseudopregnancy model was performed in a viviparous lizard, Barisia imbricata imbricata, to determine whether the absence of embryos would affect the pattern of progesterone production or the corpus luteum histology. Blood samples were obtained prior to ovulation and at 8, 16, and 24 weeks after ovulation (pseudopregnant and pregnant lizards), as well as one day after parturition (pregnant lizards) or 32 weeks after ovulation (pseudopregnant lizards). The corpus luteum was surgically removed one day after blood samples were obtained. Blood aliquots from nongravid females were obtained at similar timepoints. We found a significant reduction in plasma progesterone concentrations at 24 and 32 weeks post-ovulation in pseudopregnant lizards compared with those observed at similar times in intact pregnant lizards, whereas the progesterone levels in non-gestant lizards remained significantly lower than in either pseudopregnant or pregnant lizards. Moreover, we observed that the histological appearance of the corpus luteum from pseudogestational females (obtained 24 and 32 weeks post-ovulation) differed from the corpora lutea from lizards in late gestation and intact parturient lizards. These observations suggest that the conceptus participates in the regulation of progesterone production in late gestation and also in luteolysis control.

  3. Progesterone and peripheral nerve regeneration

    Institute of Scientific and Technical Information of China (English)

    Fei Fan; Haichao Li; Yuwei Wang; Yanglin Zheng; Lianjun Jia; Zhihui Wang

    2006-01-01

    OBJECTIVE: To explore the effect of progesterone on peripheral nerve regeneration.DATA SOURCES: An online search of Medline and OVID databases was under taken to identify articles about progesterone and peripheral nerve regeneration published in English between January 1990 and June 2004 by using the keywords of "peripheral nerve, injury, progesterone, regeneration".STUDY SELECTION: The data were primarily screened, those correlated with progesterone and peripheral nerve regeneration were involved, and their original articles were further searched, the repetitive studies or reviews were excluded.DATA EXTRACTION: Totally 59 articles about progesterone and peripheral nerve regeneration were collected, and 26 of them were involved, the other 33 excluded ones were the repetitive studies or reviews.DATA SYNTHESIS: Recent researches found that certain amount of progesterone could be synthetized in peripheral nervous system, and the expression of progesterone receptor could be found in sensory neurons and Schwann cells. After combined with the receptor, endogenous and exogenous progesterone can accelerate the formation of peripheral nerve myelin sheath, also promote the axonal regeneration.CONCLUSION: Progesterone plays a role in protecting neurons, increasing the sensitivity of nerve tissue to nerve growth factor, and accelerating regeneration of nerve in peripheral nerve regeneration, which provides theoretical references for the treatment of demyelinated disease and nerve injury, as well as the prevention of neuroma, especially that the in vivo level of progesterone should be considered for the elderly people accompanied by neuropathy and patients with congenital luteal phase defect, which is of positive significance in guiding the treatment.

  4. Intrauterine devices containing progesterone.

    Science.gov (United States)

    Murad, F

    1977-05-01

    Characteristics of progesterone-releasing IUDs are reported. At present, the only progesterone-containing IUD on the market is Progestasert, a T-shaped ethylene vinyl acetate copolymer device containing 38 mg progesterone in silicone. The device releases approximately 65 mcg/day into the uterine cavity over the course of 1-year. The device does not alter pituitary function or ovulation, nor does it depend on a local mechanical effect. Rather, it may exert its effect by inhibiting sperm capacitation or survival, or it may prevent nidation by alterning the endometrium. The reported pregnancy rate for Progestasert is 1.9% in parous women and 2.5% in nulliparous women. This efficacy rate is similar to that for other IUDs and low-dose progestin-only oral contraceptives. Breakthrough bleeding is the most common side effect, and perhaps 10-15% of the acceptors will have the device removed for either bleeding, pain, or infection. The rate of spontaneous expulsion of the device is about 3-8%. It is recommended that the device be inserted during or shortly after the menstrual period.

  5. Determination of molecular species of lecithin from erythrocytes and plasma

    NARCIS (Netherlands)

    Golde, L.M.G. van; Tomasi, V.; Deenen, L.L.M. van

    The molecular species of lecithin from erythrocyte and plasma of man and rabbit were determined after conversion of the lecithins into diglycerides by means of hydrolysis with phospholipase C. The resultant diglycerides were separated by thin-layer chromatography on silica impregnated with silver

  6. Determination of molecular species of lecithin from erythrocytes and plasma

    NARCIS (Netherlands)

    Golde, L.M.G. van; Tomasi, V.; Deenen, L.L.M. van

    1967-01-01

    The molecular species of lecithin from erythrocyte and plasma of man and rabbit were determined after conversion of the lecithins into diglycerides by means of hydrolysis with phospholipase C. The resultant diglycerides were separated by thin-layer chromatography on silica impregnated with silver ni

  7. Altrenogest and progesterone therapy during pregnancy in bottlenose dolphins (Tursiops truncatus) with progesterone insufficiency.

    Science.gov (United States)

    Robeck, Todd R; Gill, Claudia; Doescher, Bethany M; Sweeney, Jay; De Laender, Piet; Van Elk, Cornelis E; O'Brien, Justine K

    2012-06-01

    Progesterone production is essential for growth and development of the conceptus during pregnancy. Abnormal development of the corpus luteum (CL) after conception can result in early embryonic loss or fetal abortion. Routine monitoring of bottlenose dolphin (Tursiops truncatus) pregnancy after artificial insemination or natural conception with ultrasonography and serum progesterone determination has allowed for the establishment of expected fetal growth rates and hormone concentrations. Using these monitoring techniques, we revealed four pregnant dolphins (12-24 yr old) with abnormally low progesterone production indicative of luteal insufficiency. Once diagnosed, animals were placed on altrenogest (0.044-0.088 mg/kg once daily) alone or with oral progesterone (50-200 mg twice daily). Doses of hormone were increased or decreased in each animal based on how fetal skull biparietal and thoracic growth rates compared with published normal values. Hormones were withdrawn starting from day 358 of gestation in animals 1 and 2, with labor occurring 6 and 7 days after withdrawal and at 376 and 373 days of gestation, respectively. Both deliveries were dystocic, with each calf requiring manual extraction and fetotomy for calf 1. The fetuses in animals 3 and 4 died at 348 and 390 days of gestation, respectively. Induction of labor was attempted in both animals, after fetal death, by using a combination of rapid progesterone withdrawal and steroid and prostaglandin F2alpha administration. The calf of animal 4 had to be removed with manual cervical dilation and fetotomy All adult females survived the procedures. These data provide the first in vivo evidence that the CL is the primary source of progesterone throughout pregnancy in the bottlenose dolphin. Until further characterization of hormones required during pregnancy and at parturition has been accomplished, the exogenous progestagen supplementation protocol described here cannot be recommended for treatment of progesterone

  8. Spectrophotometric and Refractometric Determination of Total Protein in Avian Plasma

    Directory of Open Access Journals (Sweden)

    Rodica Căpriță

    2013-10-01

    Full Text Available The aim of this study was to compare the total protein values obtained in heparin plasma of chickens by a spectrophotometric technique (biuret method, and the values obtained on the same day in the same samples by refractometry. The results obtained by refractometry (average value 2.638±0.153g% were higher than those obtained by the spectrophotometric method (average value 2.441±0.181g%. There was a low correlation (r = 0.6709 between the total protein values, determined with both methods. Protein is the major determinant of plasma refractive index, but glucose contributes too. The refractometric method is not recommended in chickens for the determination of total protein, because avian blood glucose concentration averages about twice than in mammalian blood.

  9. Determination of actarit from human plasma for bioequivalence studies.

    Science.gov (United States)

    Loya, P; Saraf, M N

    2010-11-01

    An analytical method based on high-performance liquid chromatography with ultraviolet detection (245 nm) was developed for the determination of actarit in human plasma. Coumarin was used as an internal standard. Chromatographic separation was achieved with a C8 column using a mobile phase of methanol: 1% acetic acid (50-50, v/v) with a flow rate of 1.0 ml/min. The calibration curve was linear over the range of 0.1-4.0 μg/ml (r(2) > 0.99) and the lower limit of quantification was 0.1 μg/ml. The method was validated for sensitivity, accuracy, precision, recovery and stability. The method was used to determine the concentration-time profiles of actarit in the plasma following oral administration of 100 mg actarit tablets.

  10. New validated method for piracetam HPLC determination in human plasma.

    Science.gov (United States)

    Curticapean, Augustin; Imre, Silvia

    2007-01-10

    The new method for HPLC determination of piracetam in human plasma was developed and validated by a new approach. The simple determination by UV detection was performed on supernatant, obtained from plasma, after proteins precipitation with perchloric acid. The chromatographic separation of piracetam under a gradient elution was achieved at room temperature with a RP-18 LiChroSpher 100 column and aqueous mobile phase containing acetonitrile and methanol. The quantitative determination of piracetam was performed at 200 nm with a lower limit of quantification LLQ=2 microg/ml. For this limit, the calculated values of the coefficient of variation and difference between mean and the nominal concentration are CV%=9.7 and bias%=0.9 for the intra-day assay, and CV%=19.1 and bias%=-7.45 for the between-days assay. For precision, the range was CV%=1.8/11.6 in the intra-day and between-days assay, and for accuracy, the range was bias%=2.3/14.9 in the intra-day and between-days assay. In addition, the stability of piracetam in different conditions was verified. Piracetam proved to be stable in plasma during 4 weeks at -20 degrees C and for 36 h at 20 degrees C in the supernatant after protein precipitation. The new proposed method was used for a bioequivalence study of two medicines containing 800 mg piracetam.

  11. Spectrofluorimetric determination of amlodipine in human plasma without derivatization

    Directory of Open Access Journals (Sweden)

    Yucel Kadioglu

    2012-12-01

    Full Text Available A rapid and sensitive spectrofluorimetric method was developed for the determination of amlodipine (AD, a calcium channel blocker, in the plasma. The type of solvent, the wavelength range, and the range of AD concentration were selected to optimize the experimental conditions. The calibration curves were linear (r² >0.997 in the concentration range of 0.1-12.5 ppm of AD. The limit of quantitation and limit of detection values for the method for plasma samples were 0.1 ppm and 0.07 ppm, respectively. The precision calculated as the relative standard deviation was less than 3.5%, and the accuracy (relative error was better than 5.5% (n=6. The method developed in this study can be directly and easily applied for the determination of AD in the plasma without derivatization in plasma.Método espectrofluorometrico rápido e sensível é descrito para a determinação de anlodipina (AD, um bloqueador de canais de cálcio, em amostras de plasma. O tipo de solvente, a faixa de comprimento de onda e a faixa de concentração foram escolhidas a fim de otimizar as condições experimentais. As curvas de calibração foram lineares (r > 0,997 na faixa de concentração de 0,1-12,5 ppm de AD. Os valores LoQ e LoD do método para amostras de plasma foram 0,1 ppm e 0,07 ppm, respectivamente. A precisão calculada como desvio padrão relativo (RSD foi menor do que 3,5% e a precisão (erro relativo foi melhor do que 5,5% (n=6. O método desenvolvido neste estudo pode ser fácil e diretamente aplicado para a determinação de AD sem derivatização no plasma.

  12. 国产天然孕酮阴道环的抗生育作用药效学和动物体内血药浓度变化研究%Study on Pharmacodynamics and Plasma Concentration of Progesterone Vaginal Ring

    Institute of Scientific and Technical Information of China (English)

    桂幼伦; 郭湘洁; 谢淑武; 邵海浩; 曹霖; 朱焰

    2011-01-01

    Objective: To study on long-term contraceptive effectiveness and plasma concentration and release duration of natural progesterone vaginal ring. Methods: 1) Twenty-four fertility New Zealand rabbits were randomly divided into 4 groups: high-dose progesterone vaginal ring (840 mg/ring), mid-dose progesterone vaginal ring (420 mg / ring), tings low-dose progesterone vaginal ring (210 mg/ring), blank vaginal ting. After recovering to be mating, the long-term contraceptive effects of progesterone vaginal ring on rabbit were observed. 2) Twelve castred goats were randomly divided into 2 groups and given the domestic ring and Chile ting, respectively. Blood samples were taken before and after inserting tings at different time points via jugular vein. Magnetic enzymelinked immunosorbent system was applied to assay the concentration of progesterone. Results: 1) There was no significant effect on the general condition of the animals, the contraceptive effect in high-dose group and mid-dose group was good, long performance could maintain 3 months, while the low-dose contraceptive was less effective. 2) Plasma concentration of domestic and Chilean progesterone vaginal ring except 20 d, 27 d, 90 d was not significantly different outside (P>0.05). Conclusion: Natural progesterone vaginal rings and high-dose and mid-dose groups had long-term contraceptive effect. The domestic vaginal ring of progesterone can be as a substitute of imported vaginal ring of progesterone.%目的:研究和观察天然孕酮阴道环的长效避孕效果,及其在山羊体内的血药浓度和持续释放时间.方法:①将24只具有生育能力的新西兰雌兔随机分为4组,分别阴道置入空白环、天然孕酮阴道环低剂量(210 mg/环)、中剂量(420 mg/环)和高剂量((840 mg/环).手术15 d后进行交配,观察天然孕酮阴道环对家兔的长效避孕效果.②将12只去卵巢山羊随机分为2组,分别给予国产和智利产天然孕酮阴道环,并于置入阴道环

  13. Use of refractometry for determination of psittacine plasma protein concentration.

    Science.gov (United States)

    Cray, Carolyn; Rodriguez, Marilyn; Arheart, Kristopher L

    2008-12-01

    Previous studies have demonstrated both poor and good correlation of total protein concentrations in various avian species using refractometry and biuret methodologies. The purpose of the current study was to compare these 2 techniques of total protein determination using plasma samples from several psittacine species and to determine the effect of cholesterol and other solutes on refractometry results. Total protein concentration in heparinized plasma samples without visible lipemia was analyzed by refractometry and an automated biuret method on a dry reagent analyzer (Ortho 250). Cholesterol, glucose, and uric acid concentrations were measured using the same analyzer. Results were compared using Deming regression analysis, Bland-Altman bias plots, and Spearman's rank correlation. Correlation coefficients (r) for total protein results by refractometry and biuret methods were 0.49 in African grey parrots (n=28), 0.77 in Amazon parrots (20), 0.57 in cockatiels (20), 0.73 in cockatoos (36), 0.86 in conures (20), and 0.93 in macaws (38) (Prefractometry in Amazon parrots, conures, and macaws (n=25 each, PRefractometry can be used to accurately measure total protein concentration in nonlipemic plasma samples from some psittacine species. Method and species-specific reference intervals should be used in the interpretation of total protein values.

  14. Serum/plasma methylmercury determination by isotope dilution gas chromatography-inductively coupled plasma mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Baxter, Douglas C., E-mail: douglas.baxter@alsglobal.com [ALS Scandinavia AB, Aurorum 10, 977 75 Lulea (Sweden); Faarinen, Mikko [ALS Scandinavia AB, Aurorum 10, 977 75 Lulea (Sweden); Osterlund, Helene; Rodushkin, Ilia [ALS Scandinavia AB, Aurorum 10, 977 75 Lulea (Sweden); Division of Geosciences, Lulea University of Technology, 977 87 Lulea (Sweden); Christensen, Morten [ALS Scandinavia AB, Maskinvaegen 2, 183 53 Taeby (Sweden)

    2011-09-09

    Highlights: {center_dot} We determine methylmercury in serum and plasma using isotope dilution calibration. {center_dot} Separation by gas chromatography and detection by inductively coupled plasma mass spectrometry. {center_dot} Data for 50 specimens provides first reference range for methylmercury in serum. {center_dot} Serum samples shown to be stable for 11 months in refrigerator. - Abstract: A method for the determination of methylmercury in plasma and serum samples was developed. The method uses isotope dilution with {sup 198}Hg-labeled methylmercury, extraction into dichloromethane, back-extraction into water, aqueous-phase ethylation, purge and trap collection, thermal desorption, separation by gas chromatography, and mercury isotope specific detection by inductively coupled plasma mass spectrometry. By spiking 2 mL sample with 1.2 ng tracer, measurements in a concentration interval of (0.007-2.9) {mu}g L{sup -1} could be performed with uncertainty amplification factors <2. A limit of quantification of 0.03 {mu}g L{sup -1} was estimated at 10 times the standard deviation of concentrations measured in preparation blanks. Within- and between-run relative standard deviations were <10% at added concentration levels of 0.14 {mu}g L{sup -1}, 0.35 {mu}g L{sup -1} and 2.8 {mu}g L{sup -1}, with recoveries in the range 82-110%. Application of the method to 50 plasma/serum samples yielded a median (mean; range) concentration of methylmercury of 0.081 (0.091; <0.03-0.19) {mu}g L{sup -1}. This is the first time methylmercury has been directly measured in this kind of specimen, and is therefore the first estimate of a reference range.

  15. Spectrophotometric and Refractometric Determination of Total Protein in Avian Plasma

    OpenAIRE

    2013-01-01

    The aim of this study was to compare the total protein values obtained in heparin plasma of chickens by a spectrophotometric technique (biuret method), and the values obtained on the same day in the same samples by refractometry. The results obtained by refractometry (average value 2.638±0.153g%) were higher than those obtained by the spectrophotometric method (average value 2.441±0.181g%). There was a low correlation (r = 0.6709) between the total protein values, determined with both methods...

  16. CORRELATION BETWEEN CLINICAL PATHOLOGY OF LUMINAL B BREAST CANCER AND DETERMINATION OF ESTROGEN RECEPTOR, PROGESTERONE RECEPTOR AND HER2 EXPRESSION COMBINED WITH NUCLEAR MORPHOLOGY.

    Science.gov (United States)

    Yin, D; Wang, Y L; Wang, Y F; Yang, L; Zhang, L; Tang, C; Xie, W; Ma, Y

    2015-01-01

    Breast cancer, one of the most common malignant tumors in females, draws little attention because of its untypical symptoms and signs, so the disease is usually confirmed too late, in an advanced stage. Based on the detection of nuclear morphology parameters of luminal B breast cancer, this study explored how pathological features relate to estrogen receptor (ER) and progesterone receptor (PR) expression of human epidermal growth factor receptor-2 (HER2). A quantity of 354 breast cancer specimens with follow-up records from the department of pathology in the First People’s Hospital of Nantong and the Tumor Hospital of Nantong were selected as research subjects. Nuclear parameters of specimens stained by hematoxylin and eosin were measured by imaging analysis software. It was found that breast cancer can be divided into four types, luminal B, luminal A, HER2 over-expression and basal-like type based on immunohistochemical results of three antibodies, i.e, ER, PR and HER2. A total of 113 patients (31.8%) were confirmed with luminal B breast cancer, mostly in histological stage II; the difference of nuclear morphology was of statistical significance between ER+/PR+ and ER-/PR- (Pbreast cancer, luminal B had the lowest brain metastasis rate, while HER2 over-expression subtype was found with the highest rate of lung and pleura metastasis. Besides, luminal B possessed longer disease-free survival (DFS) than basal-like (Pbreast cancer.

  17. Pulsatile gonadotropin secretion determined by frequent sampling from the intercavernous sinus of the mare: possible modulatory role of progesterone during luteolysis.

    Science.gov (United States)

    Silvia, P J; Meyer, S L; Fitzgerald, B P

    1995-08-01

    Twelve horse mares were used in a repeated-measures design consisting of 3 replicates of 4 mares each. On Day 6 following ovulation, luteolysis was initiated with an i.m. injection of prostaglandin F2 alpha (PGF2 alpha; Lutalyse, 10 mg). Either 12 (-12) or 36 (-36) h before PGF2 alpha (PRE), blood samples were collected simultaneously from the intercavernous sinus (ICS) and jugular (JUG) vein at 10-min intervals for an 8-h period. Pituitary capacity to exogenous GnRH (2 micrograms/kg BW, i.v.) was evaluated at the alternate time point within this period. Frequent sampling and GnRH challenge were repeated 36 (+36) or 60 (+60) h after PGF2 alpha (POST). Progesterone reached a nadir 48 h after PGF2 alpha, while the first significant elevation in estradiol (p < 0.05) was not detected until 156 h. PRE LH and FSH pulse frequencies in ICS samples were 0.6 +/- 0.3 and 1.8 +/- 0.8 pulses/8 h, respectively. Three high-amplitude, concurrent ICS pulses of LH and FSH were observed during this period. These were associated with elevated gonadotropin levels in the corresponding JUG samples. In contrast, low-amplitude ICS pulses at this time were predominately releases of FSH alone that were not detectable in the JUG. Following luteolysis, POST gonadotropin pulse frequencies in the ICS increased (p < 0.001) to 7.4 +/- 0.4 and 6.8 +/- 0.5 pulses/8 h for LH and FSH, respectively. Concurrent LH and FSH releases of relatively low amplitude characterized 87.7% of the POST ICS pulses, none of which were measurable in corresponding JUG samples.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. Serum/plasma methylmercury determination by isotope dilution gas chromatography-inductively coupled plasma mass spectrometry.

    Science.gov (United States)

    Baxter, Douglas C; Faarinen, Mikko; Österlund, Heléne; Rodushkin, Ilia; Christensen, Morten

    2011-09-01

    A method for the determination of methylmercury in plasma and serum samples was developed. The method uses isotope dilution with (198)Hg-labeled methylmercury, extraction into dichloromethane, back-extraction into water, aqueous-phase ethylation, purge and trap collection, thermal desorption, separation by gas chromatography, and mercury isotope specific detection by inductively coupled plasma mass spectrometry. By spiking 2 mL sample with 1.2 ng tracer, measurements in a concentration interval of (0.007-2.9) μg L(-1) could be performed with uncertainty amplification factors levels of 0.14 μg L(-1), 0.35 μg L(-1) and 2.8 μg L(-1), with recoveries in the range 82-110%. Application of the method to 50 plasma/serum samples yielded a median (mean; range) concentration of methylmercury of 0.081 (0.091; methylmercury has been directly measured in this kind of specimen, and is therefore the first estimate of a reference range.

  19. Ionosphere Plasma State Determination in Low Earth Orbit from International Space Station Plasma Monitor

    Science.gov (United States)

    Kramer, Leonard

    2014-01-01

    A plasma diagnostic package is deployed on the International Space Station (ISS). The system - a Floating Potential Measurement Unit (FPMU) - is used by NASA to monitor the electrical floating potential of the vehicle to assure astronaut safety during extravehicular activity. However, data from the unit also reflects the ionosphere state and seems to represent an unutilized scientific resource in the form of an archive of scientific plasma state data. The unit comprises a Floating Potential probe and two Langmuir probes. There is also an unused but active plasma impedance probe. The data, at one second cadence, are collected, typically for a two week period surrounding extravehicular activity events. Data is also collected any time a visiting vehicle docks with ISS and also when any large solar events occur. The telemetry system is unusual because the package is mounted on a television camera stanchion and its data is impressed on a video signal that is transmitted to the ground and streamed by internet to two off center laboratory locations. The data quality has in the past been challenged by weaknesses in the integrated ground station and distribution systems. These issues, since mid-2010, have been largely resolved and the ground stations have been upgraded. Downstream data reduction has been developed using physics based modeling of the electron and ion collecting character in the plasma. Recursive algorithms determine plasma density and temperature from the raw Langmuir probe current voltage sweeps and this is made available in real time for situational awareness. The purpose of this paper is to describe and record the algorithm for data reduction and to show that the Floating probe and Langmuir probes are capable of providing long term plasma state measurement in the ionosphere. Geophysical features such as the Appleton anomaly and high latitude modulation at the edge of the Auroral zones are regularly observed in the nearly circular, 51 deg inclined, 400 km

  20. [Determination of plasma protein binding rate of arctiin and arctigenin with ultrafiltration].

    Science.gov (United States)

    Han, Xue-Ying; Wang, Wei; Tan, Ri-Qiu; Dou, De-Qiang

    2013-02-01

    To determine the plasma protein binding rate of arctiin and arctigenin. The ultrafiltration combined with HPLC was employed to determine the plasma protein binding rate of arctiin and arctigenin as well as rat plasma and healthy human plasma proteins. The plasma protein binding rate of arctiin with rat plasma at the concentrations of 64. 29, 32.14, 16.07 mg x L(-1) were (71.2 +/- 2.0)%, (73.4 +/- 0.61)%, (78.2 +/- 1.9)%, respectively; while the plasma protein binding rate of arctiin with healthy human plasma at the above concentrations were (64.8 +/- 3.1)%, (64.5 +/- 2.5)%, (77.5 +/- 1.7)%, respectively. The plasma protein binding rate of arctigenin with rat plasma at the concentrations of 77.42, 38.71, 19.36 mg x L(-1) were (96.7 +/- 0.41)%, (96.8 +/- 1.6)%, (97.3 +/- 0.46)%, respectively; while the plasma protein binding rate of arctigenin with normal human plasma at the above concentrations were (94.7 +/- 3.1)%, (96.8 +/- 1.6)%, (97.9 +/- 1.3)%, respectively. The binding rate of arctiin with rat plasma protein was moderate, which is slightly higher than the binding rate of arctiin with healthy human plasma protein. The plasma protein binding rates of arctigenin with both rat plasma and healthy human plasma are very high.

  1. Cancer associated proteins in blood plasma: Determining normal variation.

    Science.gov (United States)

    Stenemo, Markus; Teleman, Johan; Sjöström, Martin; Grubb, Gabriel; Malmström, Erik; Malmström, Johan; Niméus, Emma

    2016-07-01

    Protein biomarkers have the potential to improve diagnosis, stratification of patients into treatment cohorts, follow disease progression and treatment response. One distinct group of potential biomarkers comprises proteins which have been linked to cancer, known as cancer associated proteins (CAPs). We determined the normal variation of 86 CAPs in 72 individual plasma samples collected from ten individuals using SRM mass spectrometry. Samples were collected weekly during 5 weeks from ten volunteers and over one day at nine fixed time points from three volunteers. We determined the degree of the normal variation depending on interpersonal variation, variation due to time of day, and variation over weeks and observed that the variation dependent on the time of day appeared to be the most important. Subdivision of the proteins resulted in two predominant protein groups containing 21 proteins with relatively high variation in all three factors (day, week and individual), and 22 proteins with relatively low variation in all factors. We present a strategy for prioritizing biomarker candidates for future studies based on stratification over their normal variation and have made all data publicly available. Our findings can be used to improve selection of biomarker candidates in future studies and to determine which proteins are most suitable depending on study design.

  2. Use of a probing pulsed magnetic field for determining plasma parameters

    Science.gov (United States)

    Rousskikh, A. G.; Oreshkin, V. I.; Zhigalin, A. S.; Yushkov, G. Yu.

    2016-11-01

    A novel, simple, and readily usable method is proposed for measuring the electrical conductivity and temperature of a plasma. The method is based on the interaction of the test plasma with a pulsed magnetic field. The electric signals induced by the magnetic field in the circuits of two probes (miniature solenoids), one immersed in the test plasma and the other placed outside the plasma, provide data for estimating the plasma parameters. The method was verified experimentally by determining the parameters of the plasma flows generated in the cathode spots high-current pulsed vacuum arcs that were used to form cylindrical shells of bismuth Z-pinch plasma.

  3. Reduced plasma total homocysteine concentrations in Type 1 diabetes mellitus is determined by increased renal clearance.

    NARCIS (Netherlands)

    Veldman, B.A.J.; Vervoort, G.M.M.; Blom, H.J.; Smits, P.

    2005-01-01

    INTRODUCTION: Elevated plasma levels of total homocysteine are related to the development of vascular complications. Patients with diabetes mellitus are particularly at risk for the development of these complications. Several factors determine plasma total homocysteine including renal function.

  4. 99mTc-albumin can replace 125I-albumin to determine plasma volume repeatedly

    DEFF Research Database (Denmark)

    Bonfils, Peter K; Damgaard, Morten; Stokholm, Knud H

    2012-01-01

    OBJECTIVE: Plasma volume assessment may be of importance in several disorders. The purpose of the present study was to compare the reliability of plasma volume measurements by technetium-labeled human serum albumin ((99m)Tc-HSA) with a simultaneously performed plasma volume determination with iod......OBJECTIVE: Plasma volume assessment may be of importance in several disorders. The purpose of the present study was to compare the reliability of plasma volume measurements by technetium-labeled human serum albumin ((99m)Tc-HSA) with a simultaneously performed plasma volume determination...

  5. Determining the resistance of X-pinch plasma

    Institute of Scientific and Technical Information of China (English)

    Zhao Shen; Xue Chuang; Zhu Xin-Lei; Zhang Ran; Luo Hai-Yun; Zou Xiao-Bing; Wang Xin-Xin

    2013-01-01

    The current and the voltage of an X-pinch were measured.The inductance of the X-pinch was assumed to be a constant and estimated by the calculation of the magnetic field based on the well-known Biot-Savart's Law.The voltage of the inductance was calculated with L.di/dt and subtracted from the measured voltage of the X-pinch.Then,the resistance of the X-pinch was determined and the following results were obtained.At the start of the current flow the resistance of the exploding wires is several tens of Ohms,one order of magnitude,higher than the metallic resistance of the wires at room temperature,and then it falls quickly to about 1 Ω,which reflects the physical processes occurring in the electrically exploding wires,i.e.,a current transition from the highly resistive wire core to the highly conductive plasma.It was shown that the inductive contribution to the voltage of the X-pinch is less than the resistive contribution.For the wires we used,the wires' material and diameter have no strong influence on the resistance of the X-pinch,which may be explained by the fact that the current flows through the plasma rather than through the metallic wire itself.As a result,the current is almost equally divided between two parallel X-pinches even though the diameter and material of the wires used for these two X-pinches are significantly different.

  6. Absence of progesterone effects on chlamydial genital infection in female guinea pigs.

    Science.gov (United States)

    Pasley, J N; Rank, R G; Hough, A J; Cohen, C; Barron, A L

    1985-01-01

    The effect of progesterone alone and in combination with estradiol was investigated in ovariectomized and gonadally intact female guinea pigs infected with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC). The course of the infection, as determined by the percentage of cells with GPIC (chlamydia) inclusions in Giemsa-stained vaginal scrapings, was not affected in animals receiving 5.0 mg of progesterone daily. Progesterone had no influence on the enhancement of infection by estradiol. In comparison with sesame oil-treated controls, infection was prolonged by four to six days (P less than .05) in animals receiving a combination of 5.0 mg of progesterone plus 1.0 microgram of estradiol or 1.0 microgram of estradiol alone each day. In ovariectomized animals, estradiol delayed the appearance of IgA antibody in genital secretions, whereas progesterone alone had no effect. Guinea pigs treated with estradiol or progesterone plus estradiol manifested an acute endometritis not observed in animals treated with progesterone alone or in controls receiving sesame oil. Although cervical ectopy, analogous to that seen in women with high levels of progesterone, was identified by histopathology in animals treated with progesterone, no enhancement of the chlamydial infection was observed.

  7. Assessment of postpartum uterine involution and progesterone profile in Nubian goats (Capra hircus

    Directory of Open Access Journals (Sweden)

    Majdi Elnaim Badawi

    2014-06-01

    Full Text Available A total number of 12 postpartum (pp Nubian goats were included in the study to measure the uterine involution by ultrasonography from day 3 to 31 pp. Coinciding with ultrasonography, blood samples were collected at every 3 days to monitor the ovarian activity by measuring plasma progesterone (P4 concentration using progesterone radio-immuno-assay (RIA. Uterine diameter (UD and uterine lumen (UL were maximum on day 3, and minimum on day 31 pp. More than 50% of uterine involution occurred between day 3 and day 14 pp. The end of uterine involution was characterized by small UD and absence of lochia in the UL. The maximum (0.87±0.4 ng/mL and minimum (0.54±0.2 ng/mL plasma P4 levels were reported on day 27 and day 7 pp, respectively. Completion of uterine involution was recorded at 22±3.3 days pp. There was a negative correlation between P4 level and uterine parameters (UD and UL. It can be concluded that ultrasonography is a reliable tool to determine uterine involution in Nubian goats.

  8. Effective noninvasive zygosity determination by maternal plasma target region sequencing.

    Directory of Open Access Journals (Sweden)

    Jing Zheng

    Full Text Available BACKGROUND: Currently very few noninvasive molecular genetic approaches are available to determine zygosity for twin pregnancies in clinical laboratories. This study aimed to develop a novel method to determine zygosity by using maternal plasma target region sequencing. METHODS: We constructed a statistic model to calculate the possibility of each zygosity type using likelihood ratios ( Li and empirical dynamic thresholds targeting at 4,524 single nucleotide polymorphisms (SNPs loci on 22 autosomes. Then two dizygotic (DZ twin pregnancies,two monozygotic (MZ twin pregnancies and two singletons were recruited to evaluate the performance of our novel method. Finally we estimated the sensitivity and specificity of the model in silico under different cell-free fetal DNA (cff-DNA concentration and sequence depth. RESULTS/CONCLUSIONS: We obtained 8.90 Gbp sequencing data on average for six clinical samples. Two samples were classified as DZ with L values of 1.891 and 1.554, higher than the dynamic DZ cut-off values of 1.162 and 1.172, respectively. Another two samples were judged as MZ with 0.763 and 0.784 of L values, lower than the MZ cut-off values of 0.903 and 0.918. And the rest two singleton samples were regarded as MZ twins, with L values of 0.639 and 0.757, lower than the MZ cut-off values of 0.921 and 0.799. In silico, the estimated sensitivity of our noninvasive zygosity determination was 99.90% under 10% total cff-DNA concentration with 2 Gbp sequence data. As the cff-DNA concentration increased to 15%, the specificity was as high as 97% with 3.50 Gbp sequence data, much higher than 80% with 10% cff-DNA concentration. SIGNIFICANCE: This study presents the feasibility to noninvasively determine zygosity of twin pregnancy using target region sequencing, and illustrates the sensitivity and specificity under various detecting condition. Our method can act as an alternative approach for zygosity determination of twin pregnancies in clinical

  9. Experimental determination of gap scaling in a plasma opening switch

    Science.gov (United States)

    Black, D. C.; Commisso, R. J.; Ottinger, P. F.; Swanekamp, S. B.; Weber, B. V.

    2000-09-01

    Experiments were performed to investigate the coupling between a ˜0.5 μs conduction-time, ˜0.5 MA conduction-current plasma opening switch (POS), and an electron-beam (e-beam) diode. Electrical diagnostics provided measurements of the voltage at the oil-vacuum insulator and at the diode as well as anode and cathode currents on the generator and load sides of the POS. These measurements were combined with a flow impedance model to determine the POS gap over a range of conduction times and e-beam diode impedances, and for two POS-to-load distances. A comparison of the inferred POS gap at peak power with the critical gap for magnetic insulation indicates that the POS gap is always saturated in both switch-limited and load-limited regimes. This POS gap-size scaling with load impedance is consistent with an opening mechanism dominated by erosion and not J×B forces.

  10. Effects of metformin treatment on luteal phase progesterone concentration in polycystic ovary syndrome

    Directory of Open Access Journals (Sweden)

    Meenakumari K.J.

    2004-01-01

    Full Text Available The causes of luteal phase progesterone deficiency in polycystic ovary syndrome (PCOS are not known. To determine the possible involvement of hyperinsulinemia in luteal phase progesterone deficiency in women with PCOS, we examined the relationship between progesterone, luteinizing hormone (LH and insulin during the luteal phase and studied the effect of metformin on luteal progesterone levels in PCOS. Patients with PCOS (19 women aged 18-35 years were treated with metformin (500 mg three times daily for 4 weeks prior to the test cycle and throughout the study period, and submitted to ovulation induction with clomiphene citrate. Blood samples were collected from control (N = 5, same age range as PCOS women and PCOS women during the late follicular (one sample and luteal (3 samples phases and LH, insulin and progesterone concentrations were determined. Results were analyzed by one-way analysis of variance (ANOVA, Duncan's test and Karl Pearson's coefficient of correlation (r. The endocrine study showed low progesterone level (4.9 ng/ml during luteal phase in the PCOS women as compared with control (21.6 ng/ml. A significant negative correlation was observed between insulin and progesterone (r = -0.60; P < 0.01 and between progesterone and LH (r = -0.56; P < 0.05 concentrations, and a positive correlation (r = 0.83; P < 0.001 was observed between LH and insulin. The study further demonstrated a significant enhancement in luteal progesterone concentration (16.97 ng/ml in PCOS women treated with metformin. The results suggest that hyperinsulinemia/insulin resistance may be responsible for low progesterone levels during the luteal phase in PCOS. The luteal progesterone level may be enhanced in PCOS by decreasing insulin secretion with metformin.

  11. Effects of circulating progesterone and insulin on early embryo development in beef heifers.

    Science.gov (United States)

    Mann, G E; Green, M P; Sinclair, K D; Demmers, K J; Fray, M D; Gutierrez, C G; Garnsworthy, P C; Webb, R

    2003-11-20

    The aims of this study were to determine the effect on early embryo development of feeding a diet formulated to enhance circulating insulin concentrations and secondly to investigate the association between early embryo development and maternal progesterone concentrations in beef heifers. The study was carried out in 32 Simmental x Holstein Friesian heifers 22-25 months of age weighing 506+/-7kg and in condition score 3.1+/-0.1. Animals were fed two diets that were isoenergetic and isonitrogenous, but that would encourage either propionate (diet A) or acetate (diet B) production in the rumen. The rationale was that propionate would induce a greater insulin release in response to feeding. Animals were fed a 50:50 mix of the two diets for 14 days at 0.8x maintenance, with straw provided ad libitum. Animals were then fed one of the experimental diets for 3 weeks prior to synchronisation of oestrus and insemination and for a further 16 days following mating. All heifers were blood sampled daily from oestrus synchronisation and eight animals on each diet underwent daily transrectal real-time ultrasonography to determine the day of ovulation. All heifers were slaughtered at Day 16 after mating. While feeding of diet A (propionic) caused a significant (Pbody condition making further increases in insulin difficult to achieve. Diet did not affect size of ovulatory follicle (DIET A: 15.1+/-0.7mm; diet B: 14.6+/-0.7mm), day of ovulation (diet A: 3.5+/-0.2 days; diet B: 3.4+/-0.2 days), mean plasma progesterone concentration (diet A: 4.7+/-0.4ng/ml; diet B: 5.2+/-0.3ng/ml), corpus luteum weight (diet A: 6.0+/-0.2g; diet B: 6.0+/-0.2g) or pregnancy rate (diet A: 81.3%; diet B: 81.3%). However, the proportion of well-elongated (>10cm) embryos on Day 16 was higher in animals fed diet A than in those fed diet B (84.6% versus 38.5%; P10cm) embryos with levels in these animals significantly higher on Days 4 and 5 than in heifers with small (embryos at slaughter. This study

  12. TReP-132 Is a Novel Progesterone Receptor Coactivator Required for the Inhibition of Breast Cancer Cell Growth and Enhancement of Differentiation by Progesterone

    Science.gov (United States)

    Gizard, Florence; Robillard, Romain; Gross, Barbara; Barbier, Olivier; Révillion, Françoise; Peyrat, Jean-Philippe; Torpier, Gérard; Hum, Dean W.; Staels, Bart

    2006-01-01

    The sex steroid progesterone is essential for the proliferation and differentiation of the mammary gland epithelium during pregnancy. In relation to this, in vitro studies using breast carcinoma T47D cells have demonstrated a biphasic progesterone response, consisting of an initial proliferative burst followed by a sustained growth arrest. However, the transcriptional factors acting with the progesterone receptor (PR) to mediate the progesterone effects on mammary cell growth and differentiation remain to be determined. Recently, it has been demonstrated that the transcriptional regulating protein of 132 kDa (TReP-132), initially identified as a regulator of steroidogenesis, is also a cell growth suppressor. Similar to progesterone-bound PR, TReP-132 acts by inducing the gene expression of the G1 cyclin-dependent kinase inhibitors p21WAF1/Cip1 (p21) and p27Kip1 (p27). The putative interaction between TReP-132 and progesterone pathways in mammary cells was therefore analyzed in the present study. Our results show that TReP-132 interacts in vitro and in T47D cells with progesterone-activated PR. TReP-132 synergizes with progesterone-bound PR to trans activate the p21 and p27 gene promoters at proximal Sp1-binding sites. Moreover, TReP-132 overexpression and knockdown, respectively, increased or prevented the induction of p21 and p27 gene expression by progesterone. As a consequence, TReP-132 knockdown also resulted in the loss of the inhibitory effects of progesterone on pRB phosphorylation, G1/S cell cycle progression, and cell proliferation. Furthermore, the knockdown of TReP-132 expression also prevented the induction of both early and terminal markers of breast cell differentiation which had been previously identified as progesterone target genes. As well, the progesterone-induced accumulation of lipid vacuoles was inhibited in the TReP-132-depleted cells. Finally, TReP-132 gene expression levels increased following progesterone treatment, indicating the

  13. Effect of GnRH and hCG on progesterone concentration and ovarian and luteal blood flow in diestrous mares.

    Science.gov (United States)

    Brito, L F C; Baldrighi, J M; Wolf, C A; Ginther, O J

    2017-01-01

    The objective of the present study was to investigate the effect of reproductive hormones (GnRH, hCG, LH and progesterone) on the regulation of corpus luteum (CL) and ovarian blood flow. Diestrous mares received a single treatment of saline, 100μg gonadorelin (GnRH), or 1500IU hCG 10days after ovulation. Plasma LH and progesterone concentrations, resistance index (RI) for ovarian artery blood-flow, and percentage of corpus luteum (CL) with color-Doppler signals of blood flow were determined immediately before treatment (hour 0) and at hours 0.25, 0.5, 1, 1.5, 2, 3, 4, 5, and 6. In the GnRH group, LH increased (Pblood-flow signals was greater at hour 0.5 in the GnRH group than in the saline group and was intermediate in the hCG group. The similarity among groups in progesterone concentration indicated that changes in progesterone were not involved in the GnRH and hCG stimulation of ovarian vascular perfusion. Effects of treatment might have been mediated through LH; however, since hCG biological activity is primarily LH-like, the differences in timing and degree of ovarian and luteal blood flow changes after GnRH or hCG administration in the present study suggest that GnRH might have a direct effect on ovarian blood vessels and vascular control.

  14. The LC resonance probe for determining local plasma density

    Energy Technology Data Exchange (ETDEWEB)

    Boris, D R; Fernsler, R F; Walton, S G, E-mail: david.boris.ctr@nrl.navy.mi [Naval Research Laboratory, Charge Particle Physics Branch-Code 6752, Plasma Physics Division, 4555 Overlook Ave. SW, Washington, DC 20375 (United States)

    2011-04-15

    We present a novel plasma diagnostic for measuring local plasma density in reactive-gas plasmas, and depositing plasmas. The diagnostic uses a network analyzer to measure the LC resonance (LCR) frequency of a parallel plate capacitor with inductive leads. The location of the LCR ({omega}{sub R}) in frequency space is then used as a measure of the plasma dielectric constant bold varepsilon{sub p} between the plates. By properly constructing the LCR probe, {omega}{sub R} can be tuned such that {omega}{sub R} >> {omega}{sub ce}, where {omega}{sub ce} is the electron-cyclotron frequency. Thus, the probe can be used in plasmas with varying degrees of magnetization while avoiding complications introduced to bold varepsilon{sub p} when {omega} is comparable to {omega}{sub ce}. Density measurements from the LCR probe are compared with Langmuir probe measurements in an electron-beam generated plasma in which density varied from 10{sup 9} to 10{sup 11} cm{sup -3}. An axial magnetic field, typically used to confine the electron beam, was varied between 0 to 300 G. The LCR probe showed good agreement with a Langmuir probe across the entire range of magnetic fields.

  15. Estrogen and progesterone receptor status determined by the Ventana ES 320 automated immunohistochemical stainer and the CAS 200 image analyzer in 236 early-stage breast carcinomas: prognostic significance.

    Science.gov (United States)

    Layfield, L J; Saria, E A; Conlon, D H; Kerns, B J

    1996-03-01

    The quantitation of estrogen and progesterone receptors (ER and PgR) has become the standard of care in the evaluation of patients with primary breast carcinoma. It has been demonstrated that ER and PgR detected by immunohistochemical methods in formalin-fixed paraffin-embedded tissue can be quantified by computerized image analysis. In this study, ER and PgR levels were determined by using an automated immunochemistry stainer (Ventana ES 320) and an image analyzer (CAS 200) in a series of 236 patients with stage I/II carcinoma of the breast. The degree of correlation of the ER and PgR levels determined by the dextran-coated charcoal method (DCC) with image analysis quantitation was high (r=0.75). The agreement between both methods was 77% for ER and 73% for PgR. Hormone receptor levels were correlated with prognosis as determined by overall survival. An ER level of 30 fmol/mg as determined by image analysis was established to stratify the patient population most effectively into favorable and unfavorable prognostic groups (P=0.003). An ER level of 20 fmol/mg for prognostic stratification reached statistical significance (P=0.03). The DCC method was not able to stratify the patients into prognostic groups at the traditionally accepted cutpoint of 10 fmol/mg (P=0.52). We conclude that when used in combination, automated immunohistochemistry and quantitative image analysis offer a favorable alternative to the DCC method in assessment of ER and PgR status in human mammary carcinoma. In addition, quantitative immunocytochemistry techniques may prove superior to the DCC method in specimens in which there is limited tumor volume (including fine-needle aspirates), stroma-rich tumors, and early-stage lesions including intraductal carcinoma.

  16. Concentrações plasmáticas de progesterona em borregas lanadas e deslanadas no período de abril a novembro, no Estado de São Paulo Plasma concentrations of progesterone in hair and wool ewe lambs from April to November, in São Paulo State

    Directory of Open Access Journals (Sweden)

    Aya Sasa

    2002-06-01

    Full Text Available Objetivou-se com este trabalho o fornecimento de informações sobre o padrão de secreção da progesterona (P4 e as características de manifestação do ciclo estral em borregas lanadas e deslanadas, durante o período de abril a novembro, no Estado de São Paulo. Amostras de sangue foram coletadas da veia jugular de 12 borregas das raças Santa Inês (SI, Romney Marsh (RM e Suffolk (SU, e as concentrações foram determinadas pelo método de radioimunoensaio utilizando-se kits comerciais. O estro foi detectado por machos vasectomizados (impregnados com tinta pó xadrez + óleo comestível na região peitoral em 31 borregas (10 SI, 11 RM e 10 SU. O estro foi determinado pelo confronto da observação de fêmeas marcadas pelos machos vasectomizados e dos valores de concentrações plasmáticas de progesterona inferiores a 1 ng/mL. Os resultados mostraram que a maioria dos ciclos estrais foi de duração normal (14 - 19 dias para todas as raças. A duração dos ciclos de duração normal das borregas SU foi inferior aos das borregas SI e RM, as quais não diferiram entre si. Não houve diferença entre as raças na duração dos ciclos curtos (The goal of this experiment was to provide information about the pattern of progesterone secretion (P4 and on the oestrus cycle characteristics in hair and wool ewe lambs, from April to November, in São Paulo State. Blood samples from jugular vein were collected from 12 ewe lambs Santa Inês (SI, Romney Marsh (RM and Suffolk (SU breeds, and the concentrations were determined by radioimmunoassay method, using commercial kits. Oestrus was detected by using vasectomized rams (fitted with raddle crayons + comestible oil on pectoral area in 31 ewe lambs (10 SI, 11 RM and 10 SU. The oestrus was determined by confronting the observation of marked females by vasectomized rams and values of plasma progesterone concentrations lower than 1 ng/mL. The results showed that the majority of oestrus cycles were of

  17. The determination of phenazone in blood plasma for obtained sistem suitable test of monitoring drug level

    OpenAIRE

    Mochamad Lazuardi

    2007-01-01

    The determining of Phenazone to human blood plasma from healthy man after separated by solid phase extraction (SPE) and spectroscopic measurements has been investigated. The objective of that research was to obtain system suitable test for determine the Phenazone level in biological fluids (human blood plasma), for new performed dosage regimented in clinical dentistry. The method can be divided into the following four steps. 1. Centrifugation the blood sample, 2. Extraction from blood plasma ...

  18. Determination of Glimepiride in Human Plasma by Liquid Chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Ho Hyun; Lee, Hee Joo [Seoul Clinical Laboratories, Seoul (Korea, Republic of); Chang, Kyu Young [Korean Biochip Society, Seoul (Korea, Republic of); Han, Sang Beom [ChungAng University, Seoul (Korea, Republic of)

    2004-01-15

    A sensitive method for quantitation of glimepiride in human plasma has been established using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Glipizide was used as an internal standard. Glimepiride and internal standard in plasma sample was extracted using diethyl etherethyl acetate (1 : 1). A centrifuged upper layer was then evaporated and reconstituted with the mobile phase of acetonitrile-5 mM ammonium acetate (60:40, pH 3.0). The reconstituted samples were injected into a C{sub 18} reversed-phase column. Using MS/MS in the multiple reaction monitoring (MRM) mode, glimepiride and glipizide were detected without severe interference from human plasma matrix. Glimepiride produced a protonated precursor ion ([M+H]{sup +}) at m/z 491 and a corresponding product ion at m/z 352. And the internal standard produced a protonated precursor ion ([M+H]{sup +}) at m/z 446 and a corresponding product ion at m/z 321. Detection of glimepiride in human plasma by the LC-ESI/MS/MS method was accurate and precise with a quantitation limit of 0.1 ng/mL. The validation, reproducibility, stability, and recovery of the method were evaluated. The method has been successfully applied to pharmacokinetic studies of glimepiride in human plasma

  19. 51Cr-EDTA plasma clearance in severe renal failure determined by one plasma sample

    DEFF Research Database (Denmark)

    Kamper, A L; Nielsen, S L

    1989-01-01

    at zero-time was derived from injected dose and body surface area. This method might provide values 1.5 ml/min below or 0.8 ml/min above the established method of 51Cr-EDTA plasma clearance, which would be acceptable for clinical purposes. It is concluded that exact plasma clearance of 51Cr-EDTA in severe......Two hundred and thirty-four measurements of standard 51Cr-EDTA plasma clearance were made in 50 patients with severe chronic renal failure. Based on these data two calculation methods were attempted using one plasma sample drawn 24 h after injection of 51Cr-EDTA. One of the methods used the 'one...... sample clearance' formulas disregarding exact time of plasma sampling. This method might provide values 3.1 ml/min below or 2.9 ml/min above the established method of total 51Cr-EDTA plasma clearance, and would thus provide insufficient agreement. In the other method an estimate of plasma activity...

  20. Enhanced food intake by progesterone-treated female rats is related to changes in neuropeptide genes expression in hypothalamus.

    Science.gov (United States)

    Stelmańska, Ewa; Sucajtys-Szulc, Elżbieta

    2014-01-01

    Progesterone-treated females eat more food, but the mechanism underlying this effect is not well understood. The aim of the study was to analyse the effect of progesterone on neuropeptide genes expression in rat hypothalamus. Experiments were carried out on female and male Wistar rats. Animals were treated with progesterone (100 mg per rat) for 28 days. NPY and CART mRNA levels in hypothalamus were quantified by real-time PCR. The serum progesterone concentration was determined by radioimmunoassay. Progesterone administration to females caused an increase in food intake, body mass, and white adipose tissue mass. Elevated circulating progesterone concentration up-regulated NPY and down-regulated CART genes expression in hypothalamus of females. In males, elevated blood progesterone concentration had no effect on food intake, body and fat mass and on the neuropeptide genes expression in hypothalamus. Moreover, administration of progesterone in females resulted in decrease of PR mRNA level in hypothalamus. No effect of progesterone administration on PR mRNA level in hypothalamus of males was found. The changes in neuropeptide genes expression in hypothalamus may lead to stimulation of appetite and might explain the observed increase in food intake, body and adipose tissue mass in progesterone-treated females.

  1. Determination of activated plasma fibronectin using radioactive labelled collagen I

    DEFF Research Database (Denmark)

    Fenger, M

    1984-01-01

    The plasma concentration of biological active fibronectin was assayed by a protein binding assay using 125I-collagen I as ligand and heparin as activator. The standard curve is linear for a fibronectin range of 1.1-11 pmol (0.5-5.0 micrograms) and the coefficient of variation was less than 10%. T...

  2. Determination of activated plasma fibronectin using radioactive labelled collagen I

    DEFF Research Database (Denmark)

    Fenger, M

    1984-01-01

    The plasma concentration of biological active fibronectin was assayed by a protein binding assay using 125I-collagen I as ligand and heparin as activator. The standard curve is linear for a fibronectin range of 1.1-11 pmol (0.5-5.0 micrograms) and the coefficient of variation was less than 10...

  3. A State Space Model Exhibiting a Cyclic Structure with an Application to Progesterone Concentration in Cow Milk

    DEFF Research Database (Denmark)

    Hansen, Jørgen Vinsløv; Jensen, Jens Ledet; Højsgaard, Søren

    Progesterone is a hormone linked to the reproductive status of dairy cows. Hence, with the increasing availability of on-line records of the concentration of progesterone in cow milk, there is a need for new tools to analyse such data. The aim is to find techniques for better determination......-lactations with the purpose of evaluating the use of progesterone for detection of oestrus....

  4. Elevated serum progesterone/ MII oocyte ratio on the day of human chorionic gonadotropin administration can predict impaired endometrial receptivity

    OpenAIRE

    2015-01-01

    Background: Increased serum progesterone on the day of human chorionic gonadotropin administration may affect in vitro fertilization (IVF) outcome. Objective: The aim of this study was to evaluate whether progesterone elevation on the day of human chorionic gonadotropin administration is associated with poor IVF outcome. Materials and Methods: To determine the relationship between serum progesterone on the day of HCG and the outcome of IVF-embryo transfer treatment, 378 infertile patients und...

  5. Membrane progesterone receptors: evidence for neuroprotective, neurosteroid signaling and neuroendocrine functions in neuronal cells.

    Science.gov (United States)

    Thomas, Peter; Pang, Yefei

    2012-01-01

    Membrane progesterone receptors (mPRs) are novel G protein-coupled receptors belonging to the progestin and adipoQ receptor family (PAQR) that mediate a variety of rapid cell surface-initiated progesterone actions in the reproductive system involving activation of intracellular signaling pathways (i.e. nonclassical actions). The mPRs are highly expressed in the brain, but research on their neural functions has only been conducted in a single neuronal cell line, GT1-7 cells, which have negligible nuclear progesterone receptor (PR) expression. GT1-7 cells express mPRα and mPRβ on their plasma membranes which is associated with the presence of high-affinity, specific [(3)H]-progesterone receptor binding. The neurosteroid, allopregnanolone, is an effective ligand for recombinant mPRα with a relative binding affinity of 7.6% that of progesterone. Allopregnanolone acts as a potent mPR agonist on GT1-7 cells, mimicking the progesterone-induced decrease in cAMP accumulation and its antiapoptotic actions at low nanomolar concentrations. The decrease in cAMP levels is associated with rapid progesterone-induced downregulation of GnRH pulsatile secretion from perifused GT1-7 cells. The recent suggestion that mPRs are alkaline ceramidases and mediate sphingolipid signaling is not supported by empirical evidence that TNFα does not bind to mPRs overexpressed in human cells and that exogenous sphingomyelinase is ineffective in mimicking progestin actions through mPRs to induce meiotic maturation of fish oocytes. Taken together, these recent studies indicate that mPRs mediate neuroprotective effects of progesterone and allopregnanolone and are also the likely intermediaries in progesterone-induced inhibition of pulsatile GnRH secretion in GT1-7 cells. Copyright © 2012 S. Karger AG, Basel.

  6. Micro determination of plasma and erythrocyte copper by atomic absorption spectrophotometry

    Science.gov (United States)

    Blomfield, Jeanette; Macmahon, R. A.

    1969-01-01

    The free and total plasma copper and total erythrocyte copper levels have been determined by simple, yet sensitive and highly specific methods, using atomic absorption spectrophotometry. For total copper determination, the copper was split from its protein combination in plasma or red cells by the action of hydrochloric acid at room temperature. The liberated copper was chelated by ammonium pyrrolidine dithiocarbamate and extracted into n-butyl acetate by shaking and the organic extract was aspirated into the atomic absorption spectrophotometer flame. The entire procedure was carried out in polypropylene centrifuge tubes, capped during shaking. For the free plasma copper measurement the hydrochloric acid step was omitted. Removal of the plasma or erythrocyte proteins was found to be unnecessary, and, in addition, the presence of trichloracetic acid caused an appreciable lowering of absorption. Using a double-beam atomic absorption spectrophotometer and scale expansion × 10, micro methods have been derived for determining the total copper of plasma or erythrocytes with 0·1 ml of sample, and the free copper of plasma with 0·5 ml. The macro plasma copper method requires 2 ml of plasma and is suitable for use with single-beam atomic absorption spectrophotometers. With blood from 50 blood donors, normal ranges of plasma and erythrocyte copper have been determined. PMID:5776543

  7. Progesterone inhibition of oxytocin signaling in endometrium

    Directory of Open Access Journals (Sweden)

    Cecily Vauna Bishop

    2013-08-01

    Full Text Available Expression of the oxytocin receptor (OXTR in the endometrium of ruminant species is regulated by the ovarian steroids progesterone (P and estradiol (E. Near the end of the estrous cycle, long-term exposure of endometrial epithelial cells to P results in loss of genomic P receptors (PGRs, leading to an increase in E receptors (ERs. Genomic regulation of the OXTR is mediated via suppression of ER signaling by P. Upon OT binding at the plasma membrane of endometrial cells, a signaling cascade is generated stimulating release of prostaglandin F2alpha (PGF2alpha. Transport of PGF2alpha to the ovary results in release of OT by luteal cells in a positive feedback loop leading to luteal regression. This signaling cascade can be rapidly blocked by exposing endometrial cells to physiologic levels of P. This mini review will focus on the mechanisms by which P may act to block OXTR signaling and the luteolytic cascade in the ruminant endometrium, with special focus on both non-genomic signaling pathways and non-receptor actions of P at the level of the plasma membrane. While this review focuses on ruminant species, non-classical blockage of OXTR signaling may be important for fertility in women.

  8. Electrochemical determination of methylglyoxal as a biomarker in human plasma.

    Science.gov (United States)

    Chatterjee, Sanghamitra; Wen, Jiali; Chen, Aicheng

    2013-04-15

    A novel electrochemical approach for the quantitative analysis of methylglyoxal as a biomarker in human plasma has been developed. An electrochemical sensor employing a single walled carbon nanotube modified glassy carbon electrode for the sensitive detection of methylglyoxal is delineated for the first time using square wave voltammetry. This modified electrode exhibits potent and sustained electron-mediating behavior and a well-defined reduction peak in response to methylglyoxal was observed. Under optimized experimental conditions, a wide linear dynamic range, from 0.1 to 100 μM, and high sensitivity of 76.3 nA μM⁻¹ were achieved for the detection of methylglyoxal. The interfering effect of common coexisting metabolites in human whole blood has also been investigated. The developed assay was shown to be specific and sensitive for the analysis of plasma levels of methylglyoxal in healthy volunteer and diabetic patients.

  9. Determination of Some Biochemical Parameters in the Seminal Plasma of German Shepherd Dogs

    OpenAIRE

    GÜNAY, Ülgen

    2003-01-01

    In this study, we aimed to determine the levels of some biochemical parameters in the seminal plasma of German Shepherd dogs. Seven German Shepherd dogs were used as materials. A total of 35 ejaculates, five from each dog, were collected. The seminal plasma of each ejaculate was separated by centrifugation at 5000 rpm for 10 min. Biochemical analyses of the seminal plasma were performed. In conclusion, the average concentrations of total protein, calcium, phosphorus, magnesium, sodium and pot...

  10. Progesterone receptor content in endometrial carcinoma correlates with serum levels of free estradiol

    DEFF Research Database (Denmark)

    Nyholm, H C; Nielsen, Anette Lynge; Lyndrup, J;

    1993-01-01

    OBJECTIVE. To study a possible relationship between serum levels of estrogens and androgens and the tumor content of estrogen receptors and progesterone receptors in endometrial cancer. STUDY DESIGN. Fifty postmenopausal patients were included. Receptors were determined biochemically in tissue cy...

  11. Estrogen and progesterone receptors in endometrial carcinoma: comparison of immunohistochemical and biochemical analysis

    DEFF Research Database (Denmark)

    Nyholm, H C; Nielsen, Anette Lynge; Lyndrup, J;

    1993-01-01

    In 159 endometrial carcinomas, estrogen (ER) and progesterone receptors (PR) were determined biochemically by dextran-coated charcoal (DCC) assay and immunohistochemically (ICA) on frozen sections. ICA receptor content was estimated by a total histologic score (HSCORE), including all tissue...

  12. Progesterone-induced stimulation of mammary tumorigenesis is due to the progesterone metabolite, 5α-dihydroprogesterone (5αP) and can be suppressed by the 5α-reductase inhibitor, finasteride.

    Science.gov (United States)

    Wiebe, John P; Rivas, Martin A; Mercogliano, Maria F; Elizalde, Patricia V; Schillaci, Roxana

    2015-05-01

    Progesterone has long been linked to breast cancer but its actual role as a cancer promoter has remained in dispute. Previous in vitro studies have shown that progesterone is converted to 5α-dihydroprogesterone (5αP) in breast tissue and human breast cell lines by the action of 5α-reductase, and that 5αP acts as a cancer-promoter hormone. Also studies with human breast cell lines in which the conversion of progesterone to 5αP is blocked by a 5α-reductase inhibitor, have shown that the in vitro stimulation in cell proliferation with progesterone treatments are not due to progesterone itself but to the metabolite 5αP. No similar in vivo study has been previously reported. The objective of the current studies was to determine in an in vivo mouse model if the presumptive progesterone-induced mammary tumorigenesis is due to the progesterone metabolite, 5αP. BALB/c mice were challenged with C4HD murine mammary cells, which have been shown to form tumors when treated with progesterone or the progestin, medroxyprogesterone acetate. Cells and mice were treated with various doses and combinations of progesterone, 5αP and/or the 5α-reductase inhibitor, finasteride, and the effects on cell proliferation and induction and growth of tumors were monitored. Hormone levels in serum and tumors were measured by specific RIA and ELISA tests. Proliferation of C4HD cells and induction and growth of tumors was stimulated by treatment with either progesterone or 5αP. The progesterone-induced stimulation was blocked by finasteride and reinstated by concomitant treatment with 5αP. The 5αP-induced tumors expressed high levels of ER, PR and ErbB-2. Hormone measurements showed significantly higher levels of 5αP in serum from mice with tumors than from mice without tumors, regardless of treatments, and 5αP levels were significantly higher (about 4-fold) in tumors than in respective sera, while progesterone levels did not differ between the compartments. The results indicate that

  13. Determination of rafoxanide and closantel in ovine plasma by high performance liquid chromatography.

    Science.gov (United States)

    Benchaoui, H A; McKellar, Q A

    1993-01-01

    A high performance liquid chromatographic method has been developed for the determination of rafoxanide and closantel in ovine plasma. Acetonitrile and chloroform were used for the extraction. The mean recoveries were 78.69% and 80.59% for rafoxanide and closantel, respectively. This method was applied to the characterization of rafoxanide plasma kinetics following oral administration of therapeutic doses to sheep.

  14. Thermographic determination of the sheath heat transmission coefficient in a high density plasma

    NARCIS (Netherlands)

    van den Berg, M. A.; Bystrov, K.; Pasquet, R.; Zielinski, J. J.; De Temmerman, G.

    2013-01-01

    Experiments were performed in the Pilot-PSI linear plasma device, to determine the sheath heat transmission coefficients in a high recycling regime under various conditions of density (1–20 × 1020 m−3) and plasma composition (H2, Ar, N2) relevant for the

  15. Thermographic determination of the sheath heat transmission coefficient in a high density plasma

    NARCIS (Netherlands)

    van den Berg, M. A.; Bystrov, K.; Pasquet, R.; Zielinski, J. J.; De Temmerman, G.

    2013-01-01

    Experiments were performed in the Pilot-PSI linear plasma device, to determine the sheath heat transmission coefficients in a high recycling regime under various conditions of density (1–20 × 1020 m−3) and plasma composition (H2, Ar, N2) relevant for the

  16. Role of nuclear progesterone receptor isoforms in uterine pathophysiology.

    Science.gov (United States)

    Patel, Bansari; Elguero, Sonia; Thakore, Suruchi; Dahoud, Wissam; Bedaiwy, Mohamed; Mesiano, Sam

    2015-01-01

    Progesterone is a key hormonal regulator of the female reproductive system. It plays a major role to prepare the uterus for implantation and in the establishment and maintenance of pregnancy. Actions of progesterone on the uterine tissues (endometrium, myometrium and cervix) are mediated by the combined effects of two progesterone receptor (PR) isoforms, designated PR-A and PR-B. Both receptors function primarily as ligand-activated transcription factors. Progesterone action on the uterine tissues is qualitatively and quantitatively determined by the relative levels and transcriptional activities of PR-A and PR-B. The transcriptional activity of the PR isoforms is affected by specific transcriptional coregulators and by PR post-translational modifications that affect gene promoter targeting. In this context, appropriate temporal and cell-specific expression and function of PR-A and PR-B are critical for normal uterine function. Relevant studies describing the role of PRs in uterine physiology and pathology (endometriosis, uterine leiomyoma, endometrial cancer, cervical cancer and recurrent pregnancy loss) were comprehensively searched using PubMed, Cochrane Library, Web of Science, and Google Scholar and critically reviewed. Progesterone, acting through PR-A and PR-B, regulates the development and function of the endometrium and induces changes in cells essential for implantation and the establishment and maintenance of pregnancy. During pregnancy, progesterone via the PRs promotes myometrial relaxation and cervical closure. Withdrawal of PR-mediated progesterone signaling triggers menstruation and parturition. PR-mediated progesterone signaling is anti-mitogenic in endometrial epithelial cells, and as such, mitigates the tropic effects of estrogen on eutopic normal endometrium, and on ectopic implants in endometriosis. Similarly, ligand-activated PRs function as tumor suppressors in endometrial cancer cells through inhibition of key cellular signaling pathways

  17. Concentração plasmática de progesterona em novilhas receptoras submetidas à administração de rbST, GnRH ou hCG no quinto dia do ciclo estral Plasma progesterone concentration in recipient heifers submitted to administration of rbST, GnRH or hCG on day five of the estrous cycle

    Directory of Open Access Journals (Sweden)

    J.F. Fonseca

    2001-08-01

    : T1-0/12 (0.0%, T2-0/13 (0.0%, T3-5/12 (41.7% and T4-10/15 (66.7%, being T1 and T2 different from T3 and T4 (P<0.05. Plasma progesterone concentration (ng/ml was determined by radioimmunoassay from blood samples collected from jugular vein on days 5, 13, 17 and 21. Thirty-two embryos, T1=8, T2=8, T3=7 and T4=9, were transferred to recipients at day 7. Only heifers not receiving embryos were collected on days 17 and 21. Differences on plasma progesterone concentration were observed only on day 13. The values were T1=5.01± 1.04, T2=4.80± 1.26, T3=6.42± 1.47and T4=11.16± 2.79, being T3 different from T1, T2 and T4, and T4 different from T1, T2 and T3 (P<0.05. No differences between pregnancy rates, T1=37.5% (3/8, T2=62.5% (5/8, T3=28.6% (2/7 e T4=33.3% (3/9, were found. These results showed that the FWDF on day 5 is capable to respond to GnRH-analog and hCG, to ovulate and forme an accessory corpus luteum, which allows for an endogenous increase in plasma progesterone concentration in heifers on day 13 of the estrous cycle, the critical period for establishment of pregnancy in bovine.

  18. Determination of the electron energy distribution function of a low temperature plasma from optical emission spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Dodt, Dirk Hilar

    2009-01-05

    The experimental determination of the electron energy distribution of a low pressure glow discharge in neon from emission spectroscopic data has been demonstrated. The spectral data were obtained with a simple overview spectrometer and analyzed using a strict probabilistic, Bayesian data analysis. It is this Integrated Data Analysis (IDA) approach, which allows the significant extraction of non-thermal properties of the electron energy distribution function (EEDF). The results bear potential as a non-invasive alternative to probe measurements. This allows the investigation of spatially inhomogeneous plasmas (gradient length smaller than typical probe sheath dimensions) and plasmas with reactive constituents. The diagnostic of reactive plasmas is an important practical application, needed e.g. for the monitoring and control of process plasmas. Moreover, the experimental validation of probe theories for magnetized plasmas as a long-standing topic in plasma diagnostics could be addressed by the spectroscopic method. (orig.)

  19. [Plasma cholesterol determination in birds--a diagnostic tool for detection of organophosphate and carbamate intoxication].

    Science.gov (United States)

    Kiesau, B; Kummerfeld, N

    1998-07-01

    An investigation was done on the clinical usefulness of the dry chemistry analyzer Vitros DT 60 II for determination of avian plasma cholinesterase. The analytical reliability of the method, evaluated by precision and accuracy, proved to be high for plasma of numerous pet and wild birds. Values of normal plasma-cholinesterase activity were established for different psittacine and European wild birds. Significant differences in physiologic plasma-cholinesterase activity were noted between closely related species as well as between juvenile and adult birds. These findings emphasize the necessity to use control values of the same species and age group for comparison. Dry chemistry plasma-cholinesterase determination can be used as a diagnostic tool for detection of organophosphate and carbamate poisonings in the majority of investigated birds.

  20. Improved LC method to determine ivermectin in plasma.

    Science.gov (United States)

    Prieto, J G; Merino, G; Pulido, M M; Estevez, E; Molina, A J; Vila, L; Alvarez, A I

    2003-03-26

    A simple, rapid and sensitive high-performance liquid chromatographic (HPLC) method has been developed to quantify Ivermectin (IVM) in plasma using an isocratic system with fluorescence detection. The method included a fast liquid phase extraction using cold methanol. HPLC separation was carried out by reversed phase chromatography with a mobile phase composed of methanol:acetonitrile:water with 0.2% acetic acid (45:50:5 v/v/v), pumped at flow rate of 2 ml min(-1). Fluorescence detection was performed at 365 nm (excitation) and 475 nm (emission). The calibration curve for IVM was linear from 0.25 to 100 ng ml(-1). The validation method yielded good results regarding linearity, precision, accuracy, specificity and recoveries. The values of the limit of detection (LOD) and limit of quantification (LOQ) were 0.032 and 0.167 ng ml(-1), respectively.

  1. Overview of progesterone profiles in dairy cows

    DEFF Research Database (Denmark)

    Blavy, P.; Derks, M.; Martin, O.

    2016-01-01

    The aim of this study was to gain a better understanding of the variability in shape and features of all progesterone profiles during oestrus cycles in cows, and to create templates for cycle shapes and features as a base for further research. Milk progesterone data from 1418 oestrus cycles, comi...

  2. Progesterone selectively increases amygdala reactivity in women.

    NARCIS (Netherlands)

    Wingen, G.A. van; Broekhoven, F. van; Verkes, R.J.; Petersson, K.M.; Backstrom, T.; Buitelaar, J.K.; Fernandez, G.S.E.

    2008-01-01

    The acute neural effects of progesterone are mediated by its neuroactive metabolites allopregnanolone and pregnanolone. These neurosteroids potentiate the inhibitory actions of gamma-aminobutyric acid (GABA). Progesterone is known to produce anxiolytic effects in animals, but recent animal studies s

  3. A new method for high-performance liquid chromatographic determination of drotaverine in plasma.

    Science.gov (United States)

    Mezei, J; Küttel, S; Szentmiklósi, P; Marton, S; Rácz, I

    1984-10-01

    A sensitive, specific high-performance liquid chromatographic procedure was developed for the determination of plasma drotaverine levels. Basic plasma samples were adjusted to pH 1.5 and extracted with chloroform. HPLC [n-heptane-dichloromethane-diethylamine (50:25:2)] on a microporous silica column, with a variable-wavelength UV detector set at 302 nm allowed the measurement of drotaverine at the 50-ng/mL level. The utility of this method for determination of drotaverine in dog and rat plasma was demonstrated.

  4. Autoimmune Progesterone Dermatitis: A Case Report

    Directory of Open Access Journals (Sweden)

    Rachana George

    2012-01-01

    Full Text Available Background. Autoimmune progesterone dermatitis is a rare cyclic premenstrual allergic reaction to progesterone produced during the luteal phase of a woman's menstrual cycle. Patients present with a variety of conditions including erythema multiforme, eczema, urticaria, angioedema, and progesterone-induced anaphylaxis. Case. Thirty-eight-year-old woman G2P2002 presents with erythema multiforme and urticarial rash one week prior to her menses starting one year after menarche. She was treated with oral contraceptive pills and the symptoms resolved. Conclusion. This is a typical case of progesterone autoimmunity. The diagnosis is based on cyclic nature of the dermatitis. This differentiates the condition from other allergies or systemic diseases with skin manifestations. Inhibition of ovulation in such cases results in decrease in progesterone secretion and prevention of symptoms.

  5. Determination of self generated magnetic field and the plasma density using Cotton Mouton polarimetry with two color probes

    Directory of Open Access Journals (Sweden)

    Joshi A.S.

    2013-11-01

    Full Text Available Self generated magnetic fields (SGMF in laser produced plasmas are conventionally determined by measuring the Faraday rotation angle of a linearly polarized laser probe beam passing through the plasma along with the interferogram for obtaining plasma density. In this paper, we propose a new method to obtain the plasma density and the SGMF distribution from two simultaneous measurements of Cotton Mouton polarimetry of two linearly polarized probe beams of different colors that pass through plasma in a direction normal to the planar target. It is shown that this technique allows us to determine the distribution of SGMF and the plasma density without doing interferometry of laser produced plasmas.

  6. Small satellite attitude determination during plasma brake deorbiting experiment

    Science.gov (United States)

    Khurshid, Osama; Selkäinaho, Jorma; Soken, Halil Ersin; Kallio, Esa; Visala, Arto

    2016-12-01

    This paper presents a study on attitude estimation during the Plasma Brake Experiment (PBE) onboard a small satellite. The PBE demands that the satellite be spun at a very high angular velocity, up to 200 deg/s, to deploy the tether using centrifugal force. The spin controller, based on purely magnetic actuation, and the PBE demands accurate attitude estimation for the successful execution of the experiment. The biases are important to be estimated onboard small satellites due to the closely integrated systems and relatively higher interference experienced by the sensors. However, bias estimation is even more important for PBE due to the presence of a high voltage unit, onboard the satellite, that is used to charge the tether and can be the source of interference. The attitude and the biases, when estimated simultaneously, results in an augmented state vector that poses a challenge to the proper tuning of process noise. The adaptation of process noise covariance has, therefore, been studied and analysed for the challenging PBE. It has been observed that adapting the process noise covariance improves the estimation accuracy during the spin-up phase. Therefore, it is very important to use adaptive process noise covariance estimation.

  7. Residual stresses determination in textured substrates for plasma sprayed coatings

    Science.gov (United States)

    Capek, J.; Pala, Z.; Kovarik, O.

    2015-04-01

    In this contribution, we have striven to respond to the desire of obtaining the residual stress tensor in the both cold-rolled and hot-rolled substrates designated for deposition of thermal coatings by plasma spraying. Residual stresses play an important role in the coating adhesion to the substrate and, as such, it is a good practice to analyse them. Prior to spraying, the substrate is often being grit blasted. Residual stresses and texture were quantitatively assessed in both virgin and grit blasted sample employing three attitudes. Firstly without taking preferred orientation into account, secondly from measurements of interplanar lattice spacings of planes with high Miller indices using MoKα radiation. And eventually, by calculating anisotropic elastic constants as a weighted average between single-crystal and X-ray elastic constants with weighting being done according to the amount of textured and isotropic material in the irradiated volume. In the ensuing verification analyses, it was established that the latter approach is suitable for materials with either very strong or very weak presence of texture.

  8. Vacuolar-ATPase (V-ATPase) Mediates Progesterone-Induced Uterine Fluid Acidification in Rats.

    Science.gov (United States)

    Karim, Kamarulzaman; Giribabu, Nelli; Muniandy, Sekaran; Salleh, Naguib

    2016-04-01

    We hypothesized that progesterone-induced decrease in uterine fluid pH involves V-ATPase. In this study, expression and functional activity of V-ATPase in uterus were investigated under progesterone influence. Ovariectomized adult female rats received subcutaneous injection of estradiol-17β (1 µg/kg/day) or progesterone (20 mg/kg/day) for 3 days or 3 days estradiol-17β followed by 3 days vehicle, progesterone, or estradiol-17β plus progesterone. Mifepristone, a progesterone receptor blocker, was concomitantly given to the rats which received progesterone. A day after last injection, rate of uterine fluid secretion, its HCO3 (-) concentration, and pH were determined via in vivo uterine perfusion in rats under anesthesia. V-ATPase inhibitor, bafilomycin, was introduced into the perfusion buffer, and changes in these parameters were observed. Expression of V-ATPase A1 and B1/2 proteins and mRNAs in uterus were quantified by Western blotting and real-time PCR, respectively. Distribution of these proteins was observed by immunohistochemistry. Our findings showed that under progesterone influence, uterine fluid secretion rate, HCO3 (-) concentration, and pH were significantly reduced. Administration of bafilomycin did not cause significant changes in fluid secretion rate; however, HCO3 (-) concentration and pH were significantly elevated. In parallel with these changes, expression of V-ATPase A1 and B1/2 proteins and mRNAs were significantly increased with these proteins highly distributed in uterine luminal and glandular epithelia. In conclusion, increased expression and functional activity of V-ATPase were most likely responsible for the decreased in uterine fluid pH observed under progesterone influence.

  9. Elevated Progesterone Levels on the Day of Oocyte Maturation May Affect Top Quality Embryo IVF Cycles

    Science.gov (United States)

    Huang, Bo; Ren, Xinling; Wu, Li; Zhu, Lixia; Xu, Bei; Li, Yufeng; Ai, Jihui; Jin, Lei

    2016-01-01

    In contrast to the impact of elevated progesterone on endometrial receptivity, the data on whether increased progesterone levels affects the quality of embryos is still limited. This study retrospectively enrolled 4,236 fresh in vitro fertilization (IVF) cycles and sought to determine whether increased progesterone is associated with adverse outcomes with regard to top quality embryos (TQE). The results showed that the TQE rate significantly correlated with progesterone levels on the day of human chorionic gonadotropin (hCG) trigger (P = 0.009). Multivariate linear regression analysis of factors related to the TQE rate, in conventional IVF cycles, showed that the TQE rate was negatively associated with progesterone concentration on the day of hCG (OR was -1.658, 95% CI: -2.806 to -0.510, P = 0.005). When the serum progesterone level was within the interval 2.0–2.5 ng/ml, the TQE rate was significantly lower (P 2.5 ng/ml. Then, we choose a progesterone level at 1.5ng/ml, 2.0 ng/ml and 2.5 ng/ml as cut-off points to verify this result. We found that the TQE rate was significantly different (P 2.0 ng/ml. In conclusion, the results of this study clearly demonstrated a negative effect of elevated progesterone levels on the day of hCG trigger, on TQE rate, regardless of the basal FSH, the total gonadotropin, the age of the woman, or the time of ovarian stimulation. These data demonstrate that elevated progesterone levels (>2.0 ng/ml) before oocyte maturation were consistently detrimental to the oocyte. PMID:26745711

  10. Evaluation of outcome of pregnancy in threatened abortion by serum progesterone levels

    Directory of Open Access Journals (Sweden)

    Rizwana Habib Kant

    2015-10-01

    Full Text Available Background: Early pregnancy maintenance depends on the progesterone production by the developing trophoblasts of the placenta and the corpus luteum of ovary. Threatened abortion has a psychological impact on patients so we need to have a tool that could predict the outcome of threatened abortion in advance. Serum progesterone level has shown to be a reliable marker of pregnancy outcome. The aim of the study was to assess the correlation between maternal serum progesterone level and pregnancy outcome in threatened abortion and to assess the role of maternal serum progesterone levels in the immediate diagnosis of pregnancy failure. Methods: This observational study was conducted in the Department of Gynaecology and Obstetrics, Lalla Ded Hospital, Government Medical College, Srinagar over a period of 1 year in 100 women presenting with threatened abortion of less than 12 weeks of gestation, having spotting but without cervical dilatation. Results: Mean maternal serum progesterone level in patients of threatened abortion who aborted was 21.5 +/- 10.4 nanograms per millilitre and patients of threatened abortion who progressed normally to the period of viability had mean maternal serum progesterone level of 41.6 +/- 10.8 nanograms per millilitre. Considering 24 nanograms per millilitre as cut off limit, 19% of patients of threatened abortion were having serum progesterone level of lesser than or equal to 24 nanograms per millilitre and 81% of patients of threatened abortion were having serum progesterone level of greater than 24 nanograms per millilitre. Conclusions: Serum progesterone level is easy and reliable assay for determination of pregnancy outcome. [Int J Reprod Contracept Obstet Gynecol 2015; 4(5.000: 1313-1318

  11. Preparation of (21-/sup 11/C)progesterone. A potential receptor binding radiotracer

    Energy Technology Data Exchange (ETDEWEB)

    Vaalburg, W.; Terpstra, J.W.; Wiegman, T.; Ishiwata, K.; Paans, A.M.J.; Woldring, M.G.

    Clinically, the in vivo determination of progesterone receptor density by PET is of interest to estimate the response to chemotherapy. Since PET, in combination with a suitable radiopharmaceutical provides a useful tool for the in vivo localisation and quantification of receptors in tissues, we decided to investigate the potential of (21-/sup 11/C)progesterone as a radiopharmaceutical. The synthesis of this compound will be discussed.

  12. High-Performance Liquid Chromatographic Method for Determination of Germacrone in Rat Plasma

    Institute of Scientific and Technical Information of China (English)

    HEHai-bing; TANGXing; CUIFu-de

    2004-01-01

    Aim A reliable high-performance liquid chromatographic (HPLC) method was developed for determination of gennacrone in rat. plasma. Methods The plasma samples were treated with acetonitrile and analyzed by HPLC with UV detection at 244 nm. Results The limit of detection was 100 ng·mL-1 for germacrone in plasma and the linear range was0.1004-15.06μg· mL-1 in plasma. The RSD of intra-day and inter-day assay was 1.87%-4.29% and 1.29%-5.15%, respectively. The recoveries of germacrone were over 95%. The endogenous substances in plasma did not show any interference in the analysis. Conclusion The method is accurate and convenient, and suitable for pharmacokinetic studies of gennacrone in rats.

  13. A flexible software design to determine the plasma boundary in Damavand tokamak

    Science.gov (United States)

    Ghadiri, Rasoul; Sadeghi, Yahya; Esteki, Mohammad Hossein

    2014-06-01

    A plasma boundary reconstruction code has been designed by using current filament method to calculate the magnetic flux and consequently plasma boundary in Damavand tokamak. Hence, a computer-based code "The Plasma Boundary Reconstruction Code in Tokamak (PBRCT)" was developed to make a graphical user interface and to speed up the plasma boundary estimation algorithm. All required tools as the plasma boundary and magnetic surface display (MSD), error display, primary conditions and modeling panel as well as a search motor to determine a good position and number of the current filaments to find a precise model have been considered. The core is a 3000 lines Matlab code and the graphical user interface is 10,000 lines in C# language.

  14. Methodology for determination of plasma cortisol in fish using Competitive Enzyme-Linked Immunosorbent Assay (ELISA)

    DEFF Research Database (Denmark)

    Velasco-Santamaría, Yohana M.; Cruz-Casallas, Pablo E.

    2007-01-01

    Objective. To determine plasma cortisol procedure in fish using competitive enzymelinked immunosorbent assay (ELISA). Materials and methods. Two plasma samples of juveniles rainbow trout Oncorhynchus mykiss were analized by using ELISA human kit for cortisol assay. For standard curve calibration...... seven standard solutions of cortisol in human plasma (0, 20, 50, 100, 200, 400 and 800 ng.ml-1) were used. For the recovery test 50, 100 and 200 ng.ml-1 standard solutions of cortisol were used; for the linearity test four dilutions of the fish plasma samples (1/2, 1/4, 1/8 and 1/16) were prepared....... To each well fish plasma samples and standard solutions were added and its content were conjugated to peroxidase and subsequently enzyme substrate was added. The enzymatic reaction was stopped by addition of phosphoric acid 0.5 M and the absorbance was read at 450 nm. The accuracy of the pipetting...

  15. Efeitos do estresse térmico nas concentrações plasmáticas de progesterona (P4 e estradiol 17-b (E2 e temperatura retal em cabras da raça Pardo Alpina Effects of heat stress on progesterone (P4 and estradiol-17b plasma concentrations and rectal temperature of Alpine Brown goats

    Directory of Open Access Journals (Sweden)

    Luis Fernando Uribe-Velásquez

    2001-04-01

    Full Text Available Seis cabras lactantes foram distribuídas aleatoriamente em um delineamento experimental em "crossover", em dois grupos: sob condições termoneutras e estresse térmico. Um período de adaptação de 28 dias foi seguido por quatro períodos de 14 dias cada, quando os animais sob estresse térmico foram expostos à temperatura média de 33,84ºC; THI de 86,20; BGT de 36,18 e BT de 32,11ºC das 8 às 17 horas, incluindo radiação solar simulada das 10 às 15 horas. Não houve diferença entre as concentrações plasmáticas de progesterona, mas as fêmeas submetidas ao estresse térmico apresentaram diminuição nas concentrações plasmáticas de estradiol, quando comparados ao grupo termoneutro. A temperatura retal dos animais sob estresse térmico foi mais elevada quando foi comparada à do grupo de animais em condições de termoneutralidade. As cabras mantiveram as concentrações plasmáticas da progesterona, com diminuição na secreção de estradiol, quando expostas a um estresse repetido e intermitente, a despeito de ocorrer hipertermia durante o estresse pelo calor.Six lactating goats were randomly assigned to a crossover experimental design in two groups, under thermoneutral and heat stress conditions. An adaptation period of 28 days were followed by 4-periods of 14 days each, when the animals under heat stress were exposed to an average temperature of 33.34ºC; THI of 86.20; BGT of 36.18 and BT of 32.11ºC from 8 to 17 hours, including simulated solar radiation from 10 to 15 hours. There was no difference for progesterone plasma concentrations but the animals under heat stress showed a reduction of estradiol plasma concentrations as compared to the thermoneutral group. The rectal temperature of the animals under heat stress was higher when compared to the animals under thermoneutral conditions. The goats maintained progesterone plasma concentrations with reduction of estradiol secretion when exposed to repeat stress and intermittent

  16. Accuracy of plasma turbidity measurement for determining fat intolerance during total parenteral nutrition.

    Science.gov (United States)

    Nordenström, J; Thörne, A; Lindholm, M

    1990-06-01

    The accuracy of plasma turbidity measurements in predicting ability to metabolise intravenous fat emulsions during total parenteral nutrition was studied in 35 adult surgical patients. Plasma turbidity, expressed as a light scattering index (LSI), was determined by nephelometry and compared with measured triglyceride (TG) concentrations. A poor coefficient of correlation was found between LSI and TG (r = 0.52). The sensitivity and specificity of LSI in predicting TG concentration were 19% and 96% respectively. This indicates that the measurement of LSI is more useful in ruling out hypertriglyceridaemia than in detecting it. Consequently, clinical tolerance of intravenous fat emulsion cannot be monitored by measuring plasma turbidity. In order to avoid metabolic complications which may occasionally occur during intravenous nutritional therapy including fat emulsion, determination of plasma TG levels at timed intervals are recommended.

  17. Concentration of progesterone during the development of the ovulatory follicle: II. Ovarian and uterine responses.

    Science.gov (United States)

    Cerri, R L A; Chebel, R C; Rivera, F; Narciso, C D; Oliveira, R A; Amstalden, M; Baez-Sandoval, G M; Oliveira, L J; Thatcher, W W; Santos, J E P

    2011-07-01

    Two experiments evaluated the influence of altering the concentrations of progesterone during the development of the ovulatory follicle on the composition of the follicular fluid, circulating LH and PGF(2α) metabolite (PGFM), and expression of endometrial progesterone receptor and estrogen receptor-α. In both experiments, the estrous cycles were presynchronized (GnRH and progesterone insert followed by insert removal and PGF(2α) 7 d later, and GnRH after 48 h) and cows were then enrolled in 1 of 2 treatments 7 d later (study d -16): high progesterone (HP) or low progesterone (LP). In experiment 1 (n=19), cows had their estrous cycle synchronized starting on study d -9 (GnRH and progesterone insert on d -9, and insert removal and PGF(2α) on d -2). In experiment 2 (n=25), cows were submitted to the same synchronization protocol as in experiment 1, but had ovulation induced with GnRH on study d 0. In experiment 1, plasma was sampled on d -4 and analyzed for concentrations of LH; the dominant follicle was aspirated on d 0 and the fluid analyzed for concentrations of progesterone, estradiol, and free and total IGF-1. In experiment 2, follicular development and concentrations of progesterone and estradiol in plasma were evaluated until study d 16. Uterine biopsies were collected on d 12 and 16 for progesterone receptor and estrogen receptor-α protein abundance. An estradiol/oxytocin challenge for PGFM measurements in plasma was performed on d 16. In experiments 1 and 2, LP cows had lower plasma concentrations of progesterone and greater concentrations of estradiol, and had larger ovulatory follicle diameter (20.4 vs. 17.2mm) at the end of the synchronization protocol than HP cows. Concentration of LH tended to be greater for LP than HP cows (0.98 vs. 0.84 ng/mL). The dominant follicle of LP cows had greater concentration of estradiol (387.5 vs. 330.9 ng/mL) and a lower concentration of total IGF-1 (40.9 vs. 51.7 ng/mL) than that of HP cows. In experiment 2

  18. Effect of progesterone supplementation in the first week post conception on embryo survival in beef heifers.

    Science.gov (United States)

    Beltman, M E; Lonergan, P; Diskin, M G; Roche, J F; Crowe, M A

    2009-04-15

    Progesterone is essential for establishment and maintenance of pregnancy in mammals. The objective of this study was to examine the effect of elevating progesterone during the different physiological stages of early embryo development on embryo survival. Estrus was synchronized in cross-bred beef heifers (n=197, approximately 2-years old) and they were inseminated 12-18h after estrus onset (=Day 0). Inseminated heifers were randomly assigned to 1 of 3 treatments: (1) Control, n=69; (2) progesterone supplementation using a Controlled Internal Drug Release Device (CIDR) from Day 3 to 6.5, n=64; or (3) progesterone supplementation using a CIDR from Day 4.5 to 8, n=64. Body condition (BCS) and locomotion scores (scale of 1-5) were recorded for all animals. Animals with a locomotion score >/=4 (very lame) were excluded. Embryo survival rate was determined at slaughter on Day 25. Conceptus length and weight were recorded and the corpus luteum (CL) of all pregnant animals was dissected and weighed. Supplementation with exogenous progesterone increased (P<0.05) peripheral progesterone concentrations, but did not affect embryo survival rate compared with controls. Mean CL weight, conceptus length and conceptus weight were not different between treatments. There was a positive relationship (P<0.04) between the increase in progesterone concentrations from Days 3 to 6.5 and embryo survival rate in treated heifers and a similar trend existed between the increase from Days 4.5 to 8 (P<0.06). There was also a positive relationship (P<0.05) between the progesterone concentration on Day 6.5 and the embryo survival rate in treated heifers. A direct correlation was seen between locomotion score and embryo survival rate, with higher (P<0.05) early embryo survival rates in heifers with a lower locomotion score. In conclusion, supplementation with progesterone at different stages of early embryo development increased peripheral progesterone concentration and resulted in a positive

  19. [Determination of cyclohexanone concentration in the plasma separator by gas chromatography].

    Science.gov (United States)

    Huang, Min-Ju; Yan, Lin; He, Yan-Ying; Lin, Wei-Cong

    2009-09-01

    This essay is to determine the cyclohexanone concentration of the plasma separator. The compound was introduced into the GC analytical system by the carrier gas. The determination was performed by the measurement of their peak area and by the external standard method.

  20. Evaluation of the refractometric method for the determination of total protein in avian plasma or serum.

    Science.gov (United States)

    Lumeij, J T; de Bruijne, J J

    1985-07-01

    Serum total protein concentrations in pigeon blood determined with the biuret method (TPB-se) were compared with total protein concentrations in plasma (TPR-pl) and serum (TPR-se) obtained by estimation from refractive index. The refractometric method consistently yielded higher values (Prefractometric method for determination of TP in pigeon blood is not recommended.

  1. Radioimmunoassay for progesterone in bovine milk; Radioinmunoensayo para progesterona en leche bovina

    Energy Technology Data Exchange (ETDEWEB)

    Ruiz, Miriam [Instituto Superior de Ciencias y Tecnologia Nucleares, La Habana (Cuba)]. E-mail: mirian@fctn.isctn.edu.cu; Figueredo, Nancy; Castillo, Sonia; Pizarro [Centro de Isotopos, La Habana (Cuba)

    2002-07-01

    A system for the measurement of progesterone in bovine milk by radioimmunoassay has been developed and validated. This assay includes an iodine tracer purified by HPLC, the standard prepared in fat-free milk and an antibody anti-progesterone combined with second antibody. The detection limit of the assay is at 0.2 nmol/L calculated from the maximum binding menus two standard deviations and the precision is satisfactory. In the recovery assay was used 4 milk different samples and the result was 98% of recuperation. The progesterone was determinate in milk samples from post-partum animals taking samples three times per week for 40 days. The assay is simple, rapid and possibility the progesterone measurement without sample dilution, distinguish the cyclic changes of this hormone that reflect the ovarian activity in the animals. (author)

  2. The metabolic clearance of progesterone in the pregnant rat: Absence of a physiological role for the lung

    Energy Technology Data Exchange (ETDEWEB)

    Waddell, B.J.; Bruce, N.W. (Univ. of Western Australia, Nedlands)

    1989-06-01

    The metabolic clearance rate (MCR) of progesterone is among the highest for all steroid hormones studied, yet it is difficult to apportion this high MCR to specific organ contributions. The isolated lung has been shown to metabolize progesterone, and since this tissue receives the entire cardiac output, potentially it could make a major contribution to the overall MCR. This possibility was examined in the present study by measuring lung extraction of (3H)progesterone under steady-state conditions in the intact pregnant rat. Anesthetized rats (n = 6) were infused with (3H)progesterone via a femoral vein for 100 min on Day 16 of pregnancy. After the onset of steady state (40 min), four blood samples were obtained at 20-min intervals from the right ventricle and from the aorta, and the concentrations of (3H)progesterone and its metabolites were determined. Throughout the sampling period, mean arterial pressure and heart rate remained stable (two-way analysis of variance), as did the production rate (3.76 +/- 0.35 mg/day; mean +/- SEM) and the MCR (34.8 +/- 3.5 ml/min) of progesterone. Despite this high rate of clearance, there was no difference between the concentration of (3H)progesterone in arterial and right ventricular blood, indicating no net extraction of progesterone during passage through the lung. Furthermore, there was no change in the concentration of either lipid-soluble or aqueous-soluble (3H)progesterone metabolites during trans-lung passage. These observations demonstrate that the lung does not contribute to the MCR of progesterone when measured under physiological and steady-state conditions. Therefore, the relationship, MCR (ml/min) = whole-body extraction (%) x cardiac output (ml/min), is upheld for progesterone in the rat.

  3. Sensitive determination of 4-O-methylhonokiol in rabbit plasma by high performance liquid chromatography and application

    Institute of Scientific and Technical Information of China (English)

    Ming-Yue Li; Yu-Hai Tang; Xia Liu; Hai-Yan Lu; Xi-Yan Shi

    2011-01-01

    A novel high performance liquid chromatographic method was developed for the determination of 4-O- methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation. Plasma samples were treated by one-fold volume of methanol and acetoni

  4. Inhibition of Progesterone Metabolism Mimics the Effect of Progesterone Withdrawal on Forced Swim Test Immobility

    OpenAIRE

    Beckley, Ethan H.; Finn, Deborah A.

    2007-01-01

    Withdrawal from high levels of progesterone in rodents has been proposed as a model for premenstrual syndrome or postpartum depression. Forced swim test (FST) immobility, used to model depression, was assessed in intact female DBA/2J mice following progesterone withdrawal (PWD) or treatment with the 5α-reductase inhibitor finasteride. Following 5 daily progesterone injections (5 mg/kg IP) FST immobility increased only in mice withdrawn for 3 days (p < .05). In another experiment, 3 days of PW...

  5. Expression of estrogen and progesterone receptors in vestibular schwannomas and their clinical significance

    Directory of Open Access Journals (Sweden)

    Pandey Rakesh

    2009-11-01

    Full Text Available Abstract Objective The objective was to determine the expression of estrogen and progesterone receptors in vestibular schwannomas as well as to determine predictive factors for estrogen and progesterone receptor positivity. Materials and methods The study included 100 cases of vestibular schwannomas operated from January 2006 to June 2009. The clinical details were noted from the medical case files. Formaldehyde-fixed parafiin-embedded archival vestibular schwannomas specimens were used for the immunohistochemical assessment of estrogen and progesterone receptors. Results Neither estrogen nor progesterone receptors could be detected in any of our cases by means of well known immunohistochemical method using well documented monoclonal antibodies. In the control specimens, a strongly positive reaction could be seen. Conclusion No estrogen and progesterone receptor could be found in any of our 100 cases of vestibular schwannomas. Hence our study does not support a causative role of estrogen and progesterone in the growth of vestibular schwannoma as well as hormonal manipulation in the treatment of this tumor.

  6. Effects of Lipopolysaccharide and Progesterone Exposures on Embryonic Cerebral Cortex Development in Mice.

    Science.gov (United States)

    Tronnes, Ashlie A; Koschnitzky, Jenna; Daza, Ray; Hitti, Jane; Ramirez, Jan Marino; Hevner, Robert

    2016-06-01

    Our objective was to determine if progesterone pretreatment could ameliorate the detrimental effects of lipopolysaccharide (LPS)-induced inflammation on cortical neurogenesis. Timed pregnant mouse dams (n = 8) were given intraperitoneal injections of progesterone (42 mg/kg) or vehicle on embryonic day 17.5. Two hours later, mice were given intraperitoneal LPS (140 μg/kg) or vehicle. Mice were sacrificed 16 hours later on embryonic day 18. Two-color immunofluorescence was performed with primary antibodies T-box transcription factor 2 (Tbr2), ionized calcium binding adapter molecule 1 (Iba1), cleaved caspase 3 (CC3), and 5-bromo-2'-deoxyuridine (BrdU). Cells were counted, and statistical analysis was determined using analysis of variance and Tukey-Kramer method. The Tbr2 intermediate neural progenitor cell density decreased after LPS exposure (P = .0022). Pre-exposure to progesterone statistically increased Tbr2 intermediate neural progenitors compared to LPS treatment alone and was similar to controls (P = .0022). After LPS exposure, microglia displayed an activated phenotype, and cell density was increased (P < .001). Cell death rates were low among study groups but was increased in LPS exposure groups compared to progesterone alone (P = .0015). Lipopolysaccharide-induced systemic inflammation reduces prenatal neurogenesis in mice. Pre-exposure with progesterone is associated with increased neurogenesis. Progesterone may protect the preterm brain from defects of neurogenesis induced by inflammation.

  7. Effect of Monochromatic Light on Expression of Estrogen Receptor (ER) and Progesterone Receptor (PR) in Ovarian Follicles of Chicken.

    Science.gov (United States)

    Liu, Lingbin; Li, Diyan; Gilbert, Elizabeth R; Xiao, Qihai; Zhao, Xiaoling; Wang, Yan; Yin, Huadong; Zhu, Qing

    2015-01-01

    Artificial illumination is widely used in modern poultry houses and different wavelengths of light affect poultry production and behaviour. In this study, we measure mRNA and protein abundance of estrogen receptors (ERs) and progesterone receptors (PRs) in order to investigate the effect of monochromatic light on egg production traits and gonadal hormone function in chicken ovarian follicles. Five hundred and fifty-two 19-wk-old laying hens were exposed to three monochromatic lights: red (RL; 660 nm), green (GL; 560 nm), blue (BL; 480 nm) and control cool white (400-760 nm) light with an LED (light-emitting diode). There were 4 identical light-controlled rooms (n = 138) each containing 3 replicate pens (46 birds per pen). Water was supplied ad libitum and daily rations were determined according to the nutrient suggestions for poultry. Results showed that under BL conditions there was an increase in the total number of eggs at 300 days of age and egg-laying rate during the peak laying period. The BL and GL extended the duration of the peak laying period. Plasma melatonin was lowest in birds reared under BL. Plasma estradiol was elevated in the GL-exposed laying hens, and GL and BL increased progesterone at 28 wk of age. In the granulosa layers of the fifth largest preovulatory follicle (F5), the third largest preovulatory follicle (F3) and the largest preovulatory follicle (F1), ERα mRNA was increased by BL and GL. Treatment with BL increased ERβ mRNA in granulosa layers of F5, F3 and F1, while GL increased ERβ mRNA in F5 and F3. There was a corresponding increase in abundance of the proteins in the granulosa layers of F5, with an increase in PR-B, generated via an alternative splice site, relative to PR-A. Treatment with BL also increased expression of PR mRNA in all of the granulosa layers of follicles, while treatment with GL increased expression of PR mRNA in granulosa layers of SYF(small yellow follicle), F5 and F1. These results indicate that blue and green

  8. Effect of Monochromatic Light on Expression of Estrogen Receptor (ER and Progesterone Receptor (PR in Ovarian Follicles of Chicken.

    Directory of Open Access Journals (Sweden)

    Lingbin Liu

    Full Text Available Artificial illumination is widely used in modern poultry houses and different wavelengths of light affect poultry production and behaviour. In this study, we measure mRNA and protein abundance of estrogen receptors (ERs and progesterone receptors (PRs in order to investigate the effect of monochromatic light on egg production traits and gonadal hormone function in chicken ovarian follicles. Five hundred and fifty-two 19-wk-old laying hens were exposed to three monochromatic lights: red (RL; 660 nm, green (GL; 560 nm, blue (BL; 480 nm and control cool white (400-760 nm light with an LED (light-emitting diode. There were 4 identical light-controlled rooms (n = 138 each containing 3 replicate pens (46 birds per pen. Water was supplied ad libitum and daily rations were determined according to the nutrient suggestions for poultry. Results showed that under BL conditions there was an increase in the total number of eggs at 300 days of age and egg-laying rate during the peak laying period. The BL and GL extended the duration of the peak laying period. Plasma melatonin was lowest in birds reared under BL. Plasma estradiol was elevated in the GL-exposed laying hens, and GL and BL increased progesterone at 28 wk of age. In the granulosa layers of the fifth largest preovulatory follicle (F5, the third largest preovulatory follicle (F3 and the largest preovulatory follicle (F1, ERα mRNA was increased by BL and GL. Treatment with BL increased ERβ mRNA in granulosa layers of F5, F3 and F1, while GL increased ERβ mRNA in F5 and F3. There was a corresponding increase in abundance of the proteins in the granulosa layers of F5, with an increase in PR-B, generated via an alternative splice site, relative to PR-A. Treatment with BL also increased expression of PR mRNA in all of the granulosa layers of follicles, while treatment with GL increased expression of PR mRNA in granulosa layers of SYF(small yellow follicle, F5 and F1. These results indicate that blue

  9. Proteomic-Biostatistic Integrated Approach for Finding the Underlying Molecular Determinants of Hypertension in Human Plasma.

    Science.gov (United States)

    Gajjala, Prathibha R; Jankowski, Vera; Heinze, Georg; Bilo, Grzegorz; Zanchetti, Alberto; Noels, Heidi; Liehn, Elisa; Perco, Paul; Schulz, Anna; Delles, Christian; Kork, Felix; Biessen, Erik; Narkiewicz, Krzysztof; Kawecka-Jaszcz, Kalina; Floege, Juergen; Soranna, Davide; Zidek, Walter; Jankowski, Joachim

    2017-08-01

    Despite advancements in lowering blood pressure, the best approach to lower it remains controversial because of the lack of information on the molecular basis of hypertension. We, therefore, performed plasma proteomics of plasma from patients with hypertension to identify molecular determinants detectable in these subjects but not in controls and vice versa. Plasma samples from hypertensive subjects (cases; n=118) and controls (n=85) from the InGenious HyperCare cohort were used for this study and performed mass spectrometric analysis. Using biostatistical methods, plasma peptides specific for hypertension were identified, and a model was developed using least absolute shrinkage and selection operator logistic regression. The underlying peptides were identified and sequenced off-line using matrix-assisted laser desorption ionization orbitrap mass spectrometry. By comparison of the molecular composition of the plasma samples, 27 molecular determinants were identified differently expressed in cases from controls. Seventy percent of the molecular determinants selected were found to occur less likely in hypertensive patients. In cross-validation, the overall R(2) was 0.434, and the area under the curve was 0.891 with 95% confidence interval 0.8482 to 0.9349, Phypertensive patients were found to be -2.007±0.3568 and 3.383±0.2643, respectively, Phypertensives and normotensives. The identified molecular determinants may be the starting point for further studies to clarify the molecular causes of hypertension. © 2017 American Heart Association, Inc.

  10. Determining Concentrations and Temperatures in Semiconductor Manufacturing Plasmas via Submillimeter Absorption Spectroscopy

    Science.gov (United States)

    Helal, Yaser H.; Neese, Christopher F.; De Lucia, Frank C.; Ewing, Paul R.; Agarwal, Ankur; Craver, Barry; Stout, Phillip J.; Armacost, Michael D.

    2016-06-01

    Plasmas used in the manufacturing processes of semiconductors are similar in pressure and temperature to plasmas used in studying the spectroscopy of astrophysical species. Likewise, the developed technology in submillimeter absorption spectroscopy can be used for the study of industrial plasmas and for monitoring manufacturing processes. An advantage of submillimeter absorption spectroscopy is that it can be used to determine absolute concentrations and temperatures of plasma species without the need for intrusive probes. A continuous wave, 500 - 750 GHz absorption spectrometer was developed for the purpose of being used as a remote sensor of gas and plasma species. An important part of this work was the optical design to match the geometry of existing plasma reactors in the manufacturing industry. A software fitting routine was developed to simultaneously fit for the background and absorption signal, solving for concentration, rotational temperature, and translational temperature. Examples of measurements made on inductively coupled plasmas will be demonstrated. We would like to thank the Texas Analog Center of Excellence/Semiconductor Research Corporation (TxACE/SRC) and Applied Materials for their support of this work.

  11. Sensor and instrumentation for progesterone detection

    KAUST Repository

    Zia, Asif I.

    2012-05-01

    The reported research work uses a real time and noninvasive method to detect progesterone hormone concentration in purified water using Electrochemical Impedance Spectroscopy (E.I.S.) technique. Planar capacitive sensor, consisting of inter-digitated microelectrodes, is designed and fabricated on silicon substrate using thin-film Microelectromechanical system (MEMS) based semiconductor device fabrication technology. The sensor in conjunction with EIS is used to evaluate conductivity, permeability and dielectric properties of reproductive hormone progesterone and its concentration quantification in purified water. Impedance spectrums are obtained with various concentrations of the hormone in purified water by using an electric circuit in order to extract sample conductance. Relationship of sample conductance with progesterone concentration level is studied in this research work. The ability of E.I.S. to detect progesterone concentration is aimed to be used in dairy farming industry in order to obtain better reproductive performance of the dairy cattle. © 2012 IEEE.

  12. Apolipoprotein A5, a crucial determinant of plasma triglyceridelevels, is highlyresponsive to PPARalpha activators

    Energy Technology Data Exchange (ETDEWEB)

    Vu-Dac, Ngoc; Gervois, Philippe; Jakel, Heidi; Nowak, Maxine; Bauge, Eric; Dehondt, Helene; Staels, Bart; Pennacchio, Len A.; Fruchart-Najib, Jamila; Fruchart, Jean-Charles

    2003-03-30

    The recently discovered APOA5 gene has been shown in humans and mice to be important in determining plasma triglycerides (TG) levels, a major cardiovascular disease risk factor. APOAV represents the first described apolipoprotein where over-expression lowers triglyceride levels. Since fibrates represent a commonly used therapy for lowering plasma triglycerides in humans, we investigated their ability to modulate APOA5 gene expression and consequently influence plasma TG levels. Human primary hepatocytes treated with Wy 14,643 or fenofibrate displayed a strong induction of APOA5 mRNA. Deletion and mutagenesis analyses of the proximal APOA5 promoter firmly demonstrate the presence of a functional PPAR response element. These findings demonstrate that APOA5 is a highly responsive PPARa target gene and support its role as a major mediator for how fibrates reduce plasma triglycerides in humans.

  13. Determination of tolperisone enantiomers in plasma and their disposition in rats.

    Science.gov (United States)

    Yokoyama, T; Fukuda, K; Mori, S; Ogawa, M; Nagasawa, K

    1992-01-01

    A stereoselective assay for the determination of tolperisone enantiomers in plasma by high performance liquid chromatography was developed. Calibration curves obtained for the enantiomers were linear over plasma concentrations of 0.1-3.0 micrograms/ml with a detection limit of 20 ng/ml. Following intravenous bolus administration of 10 mg/kg of racemic tolperisone to rats, stereoselective disposition of tolperisone enantiomers was observed, and plasma concentrations were significantly higher for l-tolperisone than for d-tolperisone at 5, 15 and 30 min after administration. When either enantiomer was administered alone to rats, both enantiomers were found in plasma, indicating that a mutual chiral inversion occurs in the body.

  14. High-performance liquid chromatographic method for the determination of drotaverine in human plasma and urine.

    Science.gov (United States)

    Bolaji, O O; Onyeji, C O; Ogungbamila, F O; Ogunbona, F A; Ogunlana, E O

    1993-12-08

    A simple and sensitive HPLC method for the determination of drotaverine in human plasma and urine has been developed. Alkalinized plasma or urine was extracted with organic solvent and the basic components in the organic phase were back-extracted into 0.1 M HCl. An aliquot of the aqueous layer was injected onto the column and the eluent was monitored at 254 nm. Separation was performed on a C18-column with 0.02 M sodium dihydrogen phosphate-methanol (30:70, v/v) containing perchlorate ion at pH 3.2 as mobile phase. Drotaverine was well resolved from the plasma constituents and internal standard. An excellent linearity was observed between peak-height ratios and plasma concentrations and the intra- and inter-assay coefficients of variation were always drotaverine in humans as well as in animal models.

  15. Quantitative gas chromatographic determination of perfluorooctanoic acid as the benzyl ester in plasma and urine

    Energy Technology Data Exchange (ETDEWEB)

    Ylinen, M.; Hanhijaervi, H.; Peura, P.; Raemoe, O.

    1985-11-01

    A method is presented for the quantitative determination of perfluorooctanoic acid in plasma and urine of dogs. The acid was esterified by an extractive alkylation method using tetrabutylammonium ion as the counter ion and benzyl bromide as the alkylating agent. Quantitation was made by gas chromatography. Typical values for precision obtained in the determination of 5 ..mu..g/ml of perfluorooctanoic acid in plasma and urine were 3.3% (S.D. N = 6) and 3.0% (S.D. N = 6), respectively. The limit of detection was 1 ng (2.4 pmol).

  16. Modeling of progesterone-induced intracellular calcium signaling in human spermatozoa.

    Science.gov (United States)

    Li, Long-Fei; Xiang, Cheng; Zhu, Ya-Bing; Qin, Kai-Rong

    2014-06-21

    Calcium ion is a secondary messenger of mammalian spermatozoa. The dynamic change of its concentration plays a vital role in the process of sperm motility, capacitation, acrosome and fertilization. Progesterone released by the cumulus cells, as a potent stimulator of fertilization, can activate the calcium channels on the plasma membrane, which in turn triggers the dynamic change of intracellular calcium concentration. In this paper, a mathematical model of calcium dynamic response in mammalian spermatozoa induced by progesterone is proposed and numerical simulation of the dynamic model is conducted. The results show that the dynamic response of calcium concentration predicted by the model is in accordance with experimental evidence. The proposed dynamic model can be used to explain the phenomena observed in the experiments and predict new phenomena to be revealed by experimental investigations, which will provide the basis to quantitatively investigate the fluid mechanics and biochemistry for the sperm motility induced by progesterone.

  17. [Cyclic concentration of estradiol and progesterone in hysterectomized women (author's transl)].

    Science.gov (United States)

    Reynoso Ron, L; Aznar Ramos, R; Bedolla Tovar, N; Cortés-Gallegos, V

    1975-01-01

    The cyclic concentration of estradiol and progesterone was studied in 6 women, 30 years of age or less, who underwent hysterectomy, 7 to 14 months earlier, because of obstetrical complications. The basal body temperature curve, as well as the daily concentrations of plasma estradiol and progesterone during one cycle, showed a pattern similar to that of the control group, except for 2 women with a luteal phase of 16 days, who had progesterone levels of 5 ng/ml or more. Although these values are indicative of ovulation, they fell in the lower limit of the range considered as ovulatory in the laboratory that conducted the test. It is concluded that hysterectomy, with the interruption of the circulation of the ovaric branch of the uterine artery, does not affect the normal esteroidogenic function of the ovary.

  18. Oxidative stress effect on progesterone-induced blocking factor (PIBF) binding to PIBF-receptor in lymphocytes.

    Science.gov (United States)

    de la Haba, Carlos; Palacio, José R; Palkovics, Tamas; Szekeres-Barthó, Júlia; Morros, Antoni; Martínez, Paz

    2014-01-01

    Receptor-ligand binding is an essential interaction for biological function. Oxidative stress can modify receptors and/or membrane lipid dynamics, thus altering cell physiological functions. The aim of this study is to analyze how oxidative stress may alter receptor-ligand binding and lipid domain distribution in the case of progesterone-induced blocking factor/progesterone-induced blocking factor-receptor. For membrane fluidity regionalization analysis of MEC-1 lymphocytes, two-photon microscopy was used in individual living cells. Lymphocytes were also double stained with AlexaFluor647/progesterone-induced blocking factor and Laurdan to evaluate -induced blocking factor/progesterone-induced blocking factor-receptor distribution in the different membrane domains, under oxidative stress. A new procedure has been developed which quantitatively analyzes the regionalization of a membrane receptor among the lipid domains of different fluidity in the plasma membrane. We have been able to establish a new tool which detects and evaluates lipid raft clustering from two-photon microscopy images of individual living cells. We show that binding of progesterone-induced blocking factor to progesterone-induced blocking factor-receptor causes a rigidification of plasma membrane which is related to an increase of lipid raft clustering. However, this clustering is inhibited under oxidative stress conditions. In conclusion, oxidative stress decreases membrane fluidity, impairs receptor-ligand binding and reduces lipid raft clustering.

  19. The determination of micro-arc plasma composition and properties of nanoparticles formed during cathodic plasma electrolysis of 304 stainless steel

    Science.gov (United States)

    Jovović, Jovica; Stojadinović, Stevan; Vasilić, Rastko; Tadić, Nenad; Šišović, Nikola M.

    2017-05-01

    This paper presents the research focused on the determination of micro-arc plasma composition during cathodic plasma electrolysis of AISI304 stainless steel in water solution of sodium hydroxide. The complex line shape of several Fe I spectral lines was observed and, by means of a dedicated fitting procedure based on the spectral line broadening theory and H2O thermal decomposition data, the mole fraction of micro-arc plasma constituents (H2, Fe, O, H, H2O, and OH) was determined. Subsequent characterization of the cathodic plasma electrolysis product formed during the process revealed that it consists of Fe-nanoparticles with median diameter of approximately 60 nm.

  20. Levels and actions of progesterone and its metabolites in the nervous system during physiological and pathological conditions.

    Science.gov (United States)

    Melcangi, Roberto Cosimo; Giatti, Silvia; Calabrese, Donato; Pesaresi, Marzia; Cermenati, Gaia; Mitro, Nico; Viviani, Barbara; Garcia-Segura, Luis Miguel; Caruso, Donatella

    2014-02-01

    Progesterone is synthesized and actively metabolized in the central and peripheral nervous system, into neuroactive steroid metabolites, such as dihydroprogesterone, allopregnanolone and isopregnanolone. Progesterone and/or its metabolites exert a variety of effects acting as physiological regulators of neuronal and glial development and plasticity, controlling reproduction, neuroendocrine events, mood and affection. In addition, these neuroactive steroids maintain neural homeostasis and exert neuroprotective actions. In agreement, metabolic pathways of progesterone are affected by modifications in the level of gonadal hormones and by pathology or injury with a regional specificity and in a sex-dimorphic way. Therefore, observations here summarized may provide a background to design sex-specific therapies based on progesterone metabolites. On this point of view, considering that one of the major limits of a therapy based on neuroactive steroids could be modifications in their plasma levels and their consequent peripheral effects, pharmacological treatments aimed to increase their levels in the nervous system could provide an interesting therapeutic option.

  1. Metabolic level recognition of progesterone in dairy Holstein cows using probabilistic models

    Directory of Open Access Journals (Sweden)

    Ludmila N. Turino

    2014-05-01

    Full Text Available Administration of exogenous progesterone is widely used in hormonal protocols for estrous (resynchronization of dairy cattle without regarding pharmacological issues for dose calculation. This happens because it is difficult to estimate the metabolic level of progesterone for each individual cow before administration. In the present contribution, progesterone pharmacokinetics has been determined in lactating Holstein cows with different milk production yields. A Bayesian approach has been implemented to build two probabilistic progesterone pharmacokinetic models for high and low yield dairy cows. Such models are based on a one-compartment Hill structure. Posterior probabilistic models have been structurally set up and parametric probability density functions have been empirically estimated. Moreover, a global sensitivity analysis has been done to know sensitivity profile of each model. Finally, posterior probabilistic models have adequately recognized cow’s progesterone metabolic level in a validation set when Kullback-Leibler based indices were used. These results suggest that milk yield may be a good index for estimating pharmacokinetic level of progesterone.

  2. Membrane Progesterone Receptors (mPRs): Evidence for Neuroprotective, Neurosteroid Signaling and Neuroendocrine Functions in Neuronal Cells

    Science.gov (United States)

    Thomas, Peter; Pang, Yefei

    2012-01-01

    Membrane progesterone receptors (mPRs) are novel G protein-coupled receptors belonging to the progestin and adipoQ receptor family (PAQR) that mediate a variety of rapid, cell surface-initiated progesterone actions in the reproductive system involving activation of intracellular signaling pathways (i.e. nonclassical actions). The mPRs are highly expressed in the brain but research on their neural functions has only been conducted in a single neuronal cell line, GT1-7 cells, which have negligible nuclear progesterone receptor (PR) expression. GT1-7 cells express mPRα and mPRβ on their plasma membranes which is associated with the presence of high affinity, specific [3H]-progesterone receptor binding. The neurosteroid, allopregnanolone, is an effective ligand for recombinant mPRα with a relative binding affinity of 7.6% that of progesterone. Allopregnanolone acts as a potent mPR agonist on GT1-7 cells, mimicking the progesterone-induced decrease in cAMP accumulation and its antiapoptotic actions at low nanomolar concentrations.. The decrease in cAMP levels is associated with rapid progesterone-induced down-regulation of GnRH pulsatile secretion from perifused GT1-7 cells. The recent suggestion that mPRs are alkaline ceramidases and mediate sphingolipid signaling is not supported by empirical evidence that TNFα does not bind to mPRs over expressed in human cells and that exogenous sphingomyelinase is ineffective in mimicking progestin actions through mPRs to induce meiotic maturation of fish oocytes. Taken together, these recent studies indicate that mPRs mediate neuroprotective effects of progesterone and allopregnanolone and are also the likely intermediaries in progesterone-induced inhibition of pulsatile GnRH secretion in GT1-7 cells. PMID:22687885

  3. Determination of carbocysteine in human plasma by liquid chromatography/tandem mass spectrometry employing precolumn derivatization.

    Science.gov (United States)

    Chen, Xiaoyan; Zhong, Dafang; Han, Ying; Xie, Zhiyong

    2003-01-01

    A sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed to determine carbocysteine in human plasma using 2-pyridylacetic acid as the internal standard (IS). The method employed derivatization with 10 M hydrochloric acid/methanol, which significantly improved the ionization efficiency of carbocysteine. After methanol-induced protein precipitation of plasma samples, carbocysteine and the IS were derivatized and subjected to LC/MS/MS analysis using atmospheric pressure chemical ionization. The method has a lower limit of quantitation of 20 ng/mL for a 0.2-mL plasma aliquot. The intra- and inter-day precision (RSD), calculated from quality control (QC) samples, was less than 7%. The accuracy, determined using QC samples, was within +/- 1%. The method offered increased sensitivity, selectivity and speed of analysis over existing methods. The method was utilized to support clinical pharmacokinetic studies of carbocysteine in volunteers following oral administration. Copyright 2002 John Wiley & Sons, Ltd.

  4. Modulation of autoimmune rheumatic diseases by oestrogen and progesterone.

    Science.gov (United States)

    Hughes, Grant C; Choubey, Divaker

    2014-12-01

    Sexual dimorphism is evident in the risk and expression of several human autoimmune diseases. Differences in disease manifestations observed between sexes are likely to involve immunomodulation by sex steroids, nonhormonal factors encoded by genes on the X and Y chromosomes, and immunological phenomena unique to pregnancy. In systemic lupus erythematosus (SLE), and perhaps other autoantibody-mediated diseases, oestrogen seems to increase the risk of disease in genetically predisposed women by targeting key immune pathways, including the type 1 interferon (IFN) response, differentiation of CD4(+) T helper cells and survival of autoreactive B cells. By contrast, progesterone seems to reduce the risk of SLE by counteracting the effects of oestrogen on some of these same pathways, which suggests that the balance between oestrogen and progesterone can determine disease expression. In this Review we focus on the roles of the sex steroid hormones oestrogen and progesterone in modulating the risk and expression of SLE and rheumatoid arthritis. Intensive research in this area promises to identify novel therapeutic strategies and improve understanding of the immunological requirements and complications of pregnancy, and is expected to define the mechanisms behind sexual dimorphism in autoimmunity, immunity and other aspects of human health--a newly announced directive of the NIH.

  5. Bioequivalence of Progesterone Sustained Release Suppository in Rabbits

    Institute of Scientific and Technical Information of China (English)

    LONG Lihong; HUANG Qun; WU Minghui; HOU Shuxian; DAI Zongshun

    2005-01-01

    To study the bioequivalence of a kind of progesterone sustained release suppository, a randomized cross-over study was conducted in 12 rabbits. A single rectal dose of 2.75 mg/kg progesterone sustained released suppository (tested formulation, T) and progesterone suppository(reference formulation, R) was administered; a multiple dose of 2.75 mg/kg was given up to seven times with an interval of 8 h. Concentrations in serum were determined by a competitive enzyme immunoassay. The main parameters of T were: for single and multiple doses, Cmax was 48.8±11.8ng/mL and 43.5±9.4 ng/mL, Tmax was 0.5±0.3 h and 0.4±0.3 h, AUC(0-24h) was 362.4±143 ng·h·mL-1 and 310.6±70.3ng ·h·mL-1,respectively.The relative bioavailability of T to R were ( 104.2 ±13.4) % and ( 111.4 ± 19. 1 ) %, respectively. Statistical analysis showed that the two formulations were bioequivalent and T had sustained released feature.

  6. Increasing progesterone levels are associated with smoking abstinence among free-cycling women smokers who receive brief pharmacotherapy.

    Science.gov (United States)

    Saladin, Michael E; McClure, Erin A; Baker, Nathaniel L; Carpenter, Matthew J; Ramakrishnan, Viswanathan; Hartwell, Karen J; Gray, Kevin M

    2015-04-01

    Preclinical and human laboratory research suggests that (a) progesterone may decrease drug reward, craving, and smoking behavior, and (b) estradiol may enhance drug reward and smoking behavior. A modest majority of treatment research examining the relationship between menstrual cycle phase and outcomes suggests that the luteal menstrual phase, with its uniquely higher progesterone levels, is associated with better cessation outcomes. However, no studies to date have examined the effects of naturally occurring variation in progesterone and estradiol levels on medication-assisted smoking cessation. The present study sought to fill this notable gap in the treatment literature. Weekly plasma progesterone and estradiol levels were obtained from nicotine-dependent female smokers enrolled in a 4-week cessation trial. Participants (N = 108) were randomized to receive a 4-week course of either varenicline (VAR) tablets and placebo patches or placebo tablets and nicotine patches. Plasma samples were obtained 1 week before their cessation attempt and weekly during medication administration. Abstinence was assessed weekly. Weekly hormone data replicated commonly observed menstrual cycle patterns of progesterone and estradiol levels. Importantly, increases in progesterone level were associated with a 23% increase in the odds for being abstinent within each week of treatment. This effect was driven primarily by nicotine patch-treated versus VAR-treated females. This study was the first to identify an association between progesterone level (increasing) and abstinence outcomes in free-cycling women smokers who participated in a medication-based treatment. Furthermore, the potential benefits of progesterone may vary across different pharmacotherapies. Implications of these findings for smoking cessation intervention are discussed. © The Author 2015. Published by Oxford University Press on behalf of the Society for Research on Nicotine and Tobacco. All rights reserved. For

  7. Simultaneous determination of niacin, niacinamide and nicotinuric acid in human plasma.

    Science.gov (United States)

    Pfuhl, P; Kärcher, U; Häring, N; Baumeister, A; Tawab, Mona Abdel; Schubert-Zsilavecz, M

    2005-01-04

    A sensitive, specific, accurate, and reproducible HPLC/MS-method for the simultaneous quantitative determination of niacin (NA) and its main metabolites niacinamide (NAM) and nicotinuric acid (NUR) in human plasma using chinolin-3-carboxylic acid as an internal standard was developed and validated according to international guidelines for method validation. All analytes and the internal standard were separated from acidified plasma by solid phase extraction. Afterwards the extracted samples were analyzed by HPLC/MS in the positive electrospray ionization mode (ESI) and selected ion monitoring (SIM). The total run time was 7 min between injections. The assay had a lower limit of quantification of 50.0 ng/mL for each analyte using 1 mL of plasma. The calibration curves were linear in the measured range between 50.0 and 750 ng/mL plasma. The overall precision and accuracy for all concentrations of quality controls and standards was better than 15%. No indications were found for possible instabilities of niacin, niacinamide and nicotinuric acid in plasma at -20 degrees C, in the extraction solvent or after repeated thawing/freezing cycles. In stabilities were observed in whole blood and in plasma at room temperature. The recovery of the extraction method ranged from 86 to 89% for the three analytes.

  8. Determination of chlordiazepoxide, diazepam, and their major metabolites in blood or plasma by spectrophotodensitometry.

    Science.gov (United States)

    Stronjny, N; Bratin, K; Brooks, M A; de Silva, J A

    1977-07-01

    An analytical procedure was developed for the determination of chlordiazepoxide, diazepam and their major metabolites in blood or plasma. Demoxepam, a metabolite of chlordiazepoxide, is determined by spectrofluorometry after extraction. The remaining compounds are determined by spectrophotodensitometry after thin-layer chromatographic separation. The sensitivity limit of the spectrofluorometric determination of demoxepam is 0.1 to 0.2 microgram while that of the spectrophotodensitometric determination of chlordiazepoxide, diazepam and their N-desmethyl metabolites is 0.05 to 0.2 microgram. The sensitivity and specificity of the assay renders it suitable for monitoring plasma levels of chlordiazepoxide and its major metabolites following single or chronic oral administration of chlordiazepoxide hydrochloride. The sensitivity limit for diazepam and nordiazepam, its major metabolite, renders the assay useful only for the determination of plasma concentrations resulting from high dosage of diazepam. The assay was used to determine chlordiazepoxide and its metabolites following oral administration of Librium. The data showed a significant correlation to those obtained on the same specimens by differential pulse polarography and by radioimmunoassay.

  9. Quantitative Determination of Levonorgestrel in Fish Plasma using UPLC-MS/MS

    Science.gov (United States)

    In this study, a sensitive high-performance liquid chromatography electrospray tandem mass spectrometric method was developed for the determination of levonorgestrel in fish plasma using levonorgestrel-d6 as an internal standard (IS). In the laboratory, the fish cunner, (Tautogol...

  10. Betaine and folate status as cooperative determinants of plasma homocysteine in humans.

    NARCIS (Netherlands)

    Holm, P.I.; Ueland, P.M.; Vollset, S.E.; Midttun, O.; Blom, H.J.; Keijzer, M.B.; Heijer, M. den

    2005-01-01

    OBJECTIVE: Two published studies have demonstrated that betaine in the circulation is a determinant of plasma total homocysteine, but none had sufficient power to investigate the possible effect modification by folate status. METHODS AND RESULTS: We measured homocysteine, betaine, folate, vitamin B(

  11. Spectroscopic Methods for Determination of Excitation Temperatures of High-Pressure Plasma

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yu; WEN Xiaohui; ZHAN Rujuan; YANG Weihong

    2007-01-01

    A method to determine excitation temperatures based on the optical emission spectroscopy(OES)and Fermi-Dirac distribution was set up,and the temperatures of pure-argon and mixed-gases at different pressures were investigated.In this way we set up a standard process to get the excitation temperatures of plasmas operated at atmospheric and sub-atmospheric pressures.

  12. Determination of platinum in human subcellular microsamples by inductively coupled plasma mass spectrometry

    DEFF Research Database (Denmark)

    Björn, Erik; Nygren, Yvonne; Nguyen, Tam T. T. N.

    2007-01-01

    A fast and robust method for the determination of platinum in human subcellular microsamples by inductively coupled plasma mass spectrometry was developed, characterized, and validated. Samples of isolated DNA and exosome fractions from human ovarian (2008) and melanoma (T289) cancer cell lines...

  13. Determination of telmisartan in human blood plasma: Part I: Immunoassay development

    NARCIS (Netherlands)

    Hempen, C.M.; Gläsle-Schwarz, Liane; Kunz, Ulrich; Karst, U.

    2006-01-01

    Telmisartan is an angiotensin II receptor antagonist and a known drug against high blood pressure. In this report, the development of a new and rapid analytical technique, an enzyme-linked immunosorbent assay (ELISA) for the determination of telmisartan in human blood plasma is described. The

  14. Temperature dependence of the cosphi conductance in Josephson tunnel junctions determined from plasma resonance experiments

    DEFF Research Database (Denmark)

    Pedersen, Niels Falsig; Sørensen, O. H.; Mygind, Jesper

    1978-01-01

    The microwave response at 9 GHz of Sn-O-Sn tunnel-junction current biased at zero dc voltage has been measured just below the critical temperature Tc of the Sn films. The temperature dependence of the cosφ conductance is determined from the resonant response at the junction plasma frequency fp...... of the experiment....

  15. Quantitative Determination of Levonorgestrel in Fish Plasma using UPLC-MS/MS

    Science.gov (United States)

    In this study, a sensitive high-performance liquid chromatography electrospray tandem mass spectrometric method was developed for the determination of levonorgestrel in fish plasma using levonorgestrel-d6 as an internal standard (IS). In the laboratory, the fish cunner, (Tautogol...

  16. Enantioselective high-performance liquid chromatographic determination of nicardipine in human plasma.

    Science.gov (United States)

    Uno, T; Ohkubo, T; Sugawara, K

    1997-09-26

    A sensitive method for the enantioselective high-performance liquid chromatography (HPLC) determination of nicardipine in human plasma is described. (+)-Nicardipine, (-)-nicardipine and (+)-barnidipine as an internal standard are detected by an ultraviolet detector at 254 nm. Racemic nicardipine in human plasma was extracted by a rapid and simple procedure based on C18 bonded-phase extraction. The extraction samples were purified and concentrated on a pre-column using a C1 stationary phase and the enantiomers of nicardipine are quantitatively separated by HPLC on a Sumichiral OA-4500 column, containing a chemically modified Pirkle-type stationary phase. Determination of (+)- and (-)-nicardipine was possible in a concentration range of 5-100 ng ml(-1) and the limit of detection in plasma was 2.5 ng ml(-1). The recoveries of (+)- and (-)-nicardipine added to plasma were 91.4-98.4% and 93.3-96.7%, respectively, with coefficients of variation of less than 9.0 and 9.4% respectively. The method was applied to low level monitoring of (+)- and (-)-nicardipine in plasma from healthy volunteers.

  17. Progesterone modulates the LPS-induced nitric oxide production by a progesterone-receptor independent mechanism.

    Science.gov (United States)

    Wolfson, Manuel Luis; Schander, Julieta Aylen; Bariani, María Victoria; Correa, Fernando; Franchi, Ana María

    2015-12-15

    Genital tract infections caused by Gram-negative bacteria induce miscarriage and are one of the most common complications of human pregnancy. LPS administration to 7-day pregnant mice induces embryo resorption after 24h, with nitric oxide playing a fundamental role in this process. We have previously shown that progesterone exerts protective effects on the embryo by modulating the inflammatory reaction triggered by LPS. Here we sought to investigate whether the in vivo administration of progesterone modulated the LPS-induced nitric oxide production from peripheral blood mononuclear cells from pregnant and non-pregnant mice. We found that progesterone downregulated LPS-induced nitric oxide production by a progesterone receptor-independent mechanism. Moreover, our results suggest a possible participation of glucocorticoid receptors in at least some of the anti-inflammatory effects of progesterone.

  18. Determination of the concentrations of the steroids estradiol, progesterone and testosterone in bovine sera: comparison of commercial dissociation enhanced lanthanide fluorescence immunoassay kits with conventional radio and enzyme immunoassays.

    Science.gov (United States)

    Elliott, C T; Francis, K S; Shortt, H D; McCaughey, W J

    1995-06-01

    The performance of three conventional enzyme and radioimmunoassays routinely used to detect residues of anabolic steroids in cattle sera were compared with dissociation enhanced lanthanide fluorescence immunoassay (DELFIA) kits designed for the hospital market. Slight modifications to the kit reagents were required for the analysis of bovine sera. Owing to the large sample volumes used in conventional assays, detection limits were generally better than those obtained with DELFIA kits, however, assay reproducibility was enhanced using the DELFIA technology. Comparison of sera obtained from cattle implanted with anabolic steroids revealed a good correlation between alternate methods (r2 from 0.91 to 0.97). The DELFIA kits offer a faster method for measuring estradiol, progesterone and testosterone with adequate sensitivity and in a safer environment than that encountered using radioimmunoassays.

  19. "Determination of mycophenolic acid in human plasma by high-performance liquid chromatography"

    OpenAIRE

    "Mehdi Ahadi Barzoki; Mohammadreza Rouini; Kheirolla Gholami; Mahboob Lessan-Pezeshki; Saeed Rezaee

    2005-01-01

    A simple, sensitive and reproducible HPLC method is presented for determination of mycophenolic acid(MPA) in human plasma. Samples were prepared after precipitation of the plasma protein by addition of acetonitrile and naproxen was used as internal standard (I.S.). Separation was performed by reversedphase HPLC, using a Hamilton PRP-C18 Column, 51% acetonitrile and 49% potassium phosphate buffer (20 mM) at pH 3.0 as mobile phase, flow rate of 1.0 ml/min, and UV detection at 215 nm. MPA and I....

  20. Native and Reconstituted Plasma Lipoproteins in Nanomedicine: Physicochemical Determinants of Nanoparticle Structure, Stability, and Metabolism.

    Science.gov (United States)

    Pownall, Henry J; Rosales, Corina; Gillard, Baiba K; Ferrari, Mauro

    2016-09-01

    Although many acute and chronic diseases are managed via pharmacological means, challenges remain regarding appropriate drug targeting and maintenance of therapeutic levels within target tissues. Advances in nanotechnology will overcome these challenges through the development of lipidic particles, including liposomes, lipoproteins, and reconstituted high-density lipoproteins (rHDL) that are potential carriers of water-soluble, hydrophobic, and amphiphilic molecules. Herein we summarize the properties of human plasma lipoproteins and rHDL, identify the physicochemical determinants of lipid transfer between phospholipid surfaces, and discuss strategies for increasing the plasma half-life of lipoprotein- and liposome-associated molecules.

  1. Rapid determination of succinylcholine in human plasma by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Lagerwerf, A J; Vanlinthout, L E; Vree, T B

    1991-10-04

    A high-performance liquid chromatographic method with fluorometric detection has been developed for the determination of succinylcholine in human plasma. Succinylcholine shows fluorescence at 282 nm with an excitation at 257 nm. The assay is sensitive, reproducible and linear for concentrations ranging from 100 ng/ml to 100 micrograms/ml of succinylcholine. In a pilot study the plasma concentration-time curve showed a triphasic elimination, with half-lives of 0.4, 1.2 and 8 min, respectively. In a clinical setting, drugs commonly administered during anaesthesia did not interfere with the assay. This method provides a simple and time-saving alternative to existing methods.

  2. Determination of talniflumate and niflumic acid in human plasma by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Kang, Wonku; Kim, Kibum

    2009-04-01

    A simple and rapid quantification method was developed for determining both talniflumate and niflumic acid in human plasma. After simple protein precipitation with acetonitrile, the analytes were chromatographed on a reversed-phase C(18) column and detected by LC/MS/MS with electrospray ionization. The assay accuracy and precision were within the FDA guidance for the analytical method validation. This method was used to measure the plasma concentrations of both compounds from healthy subjects after a single oral dose of talniflumate, 740 mg.

  3. High Performance Liquid Chromatographic Method for Determination of Dipyridamole in Human Plasma

    Directory of Open Access Journals (Sweden)

    DAVOOD BEIGI BAND ARAB ADI

    1999-08-01

    Full Text Available A simple, rapid and specific high-performance liquid chromatographic procedure is reported for"nquantitative determination of dipyridamole in human -plasma. The assay uses a reversed-phase"nhigh-performance liquid chromatographic (HPLC and UV detection at 280nm and has a limit"nof detection of approximately 5ng/mL. The mobile phase consists of MeOH-H20 (60:40"nadjusted to pH 3.3. Dipyridamole was extracted from plasma by back-extraction procedure, with"npropranolol as the internal standard. The reproducibility of the method is satisfactory

  4. Vaginal progesterone in women with an asymptomatic sonographic short cervix in the midtrimester decreases preterm delivery and neonatal morbidity

    DEFF Research Database (Denmark)

    Romero, Roberto; Nicolaides, Kypros; Conde-Agudelo, Agustin;

    2012-01-01

    To determine whether the use of vaginal progesterone in asymptomatic women with a sonographic short cervix (≤ 25 mm) in the midtrimester reduces the risk of preterm birth and improves neonatal morbidity and mortality.......To determine whether the use of vaginal progesterone in asymptomatic women with a sonographic short cervix (≤ 25 mm) in the midtrimester reduces the risk of preterm birth and improves neonatal morbidity and mortality....

  5. Determination of fenbendazole, praziquantel and pyrantel pamoate in dog plasma by high-performance liquid chromatography.

    Science.gov (United States)

    Morovján, G; Csokán, P; Makranszki, L; Abdellah-Nagy, E A; Tóth, K

    1998-02-27

    Simple and rapid high-performance liquid chromatographic methods were developed for the determination of fenbendazole, praziquantel and pyrantel pamoate in dog plasma. The combination of these drugs is the most powerful treatment against most types of worms. Blood plasma samples obtained in a pharmacokinetic trial were prepared using solid-phase extraction. Fenbendazole and praziquantel were analyzed simultaneously by reversed-phase high-performance liquid chromatography on an octadecyl-modified silica stationary phase employing acetonitrile-phosphate buffer (pH 3.0) eluent and ultraviolet detection at 220 nm. Pyrantel was analyzed separately on a base-deactivated reversed-phase column using methanol-tetrahydrofuran-ammonium acetate buffer (pH 4.6) eluent and ultraviolet detection at 317 nm. Average recoveries for fenbendazole, praziquantel and pyrantel pamoate were 76.8, 93.4 and 90.5%, respectively. Limits of quantitation were in the range of 15-25 ng/ml plasma.

  6. A cell-free assay to determine the stoichiometry of plasma membrane proteins.

    Science.gov (United States)

    Trigo, Cesar; Vivar, Juan P; Gonzalez, Carlos B; Brauchi, Sebastian

    2013-04-01

    Plasma membrane receptors, transporters, and ion channel molecules are often found as oligomeric structures that participate in signaling cascades essential for cell survival. Different states of protein oligomerization may play a role in functional control and allosteric regulation. Stochastic GFP-photobleaching (SGP) has emerged as an affordable and simple method to determine the stoichiometry of proteins at the plasma membrane. This non-invasive optical approach can be useful for total internal reflection of fluorescence microscopy (TIRFM), where signal-to-noise ratio is very high at the plasma membrane. Here, we report an alternative methodology implemented on a standard laser scanning confocal microscope (LSCM). The simplicity of our method will allow for its implementation in any epifluorescence microscope of choice.

  7. Determination of albendazole sulfoxide in human plasma by using liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Saraner, Nihal; Özkan, Güler Yağmur; Güney, Berrak; Alkan, Erkin; Burul-Bozkurt, Nihan; Sağlam, Onursal; Fikirdeşici, Ezgi; Yıldırım, Mevlüt

    2016-06-01

    A rapid, simple and sensitive method was developed and validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for determination of albendazole sulfoxide (ABZOX) in human plasma. The plasma samples were extracted by protein precipitation using albendazole sulfoxide-d3 as internal standard (IS). The chromatographic separation was performed on Waters Xbridge C18Column (100×4.6mm, 3.5μm) with a mobile phase consisting of ammonia solution, water and methanol at a flow rate of 0.70mL/min. ABZOX was detected and identified by mass spectrometry with electrospray ionization (ESI) in positive ion and multiple-reaction monitoring (MRM) mode. The method was linear in the range of 3-1500ng/mL for ABZOX. This method was successfully applied to the bioequivalence study in human plasma samples.

  8. Magnetoresistive polyaniline-silicon carbide metacomposites: plasma frequency determination and high magnetic field sensitivity.

    Science.gov (United States)

    Gu, Hongbo; Guo, Jiang; Khan, Mojammel Alam; Young, David P; Shen, T D; Wei, Suying; Guo, Zhanhu

    2016-07-20

    The Drude model modified by Debye relaxation time was introduced to determine the plasma frequency (ωp) in the surface initiated polymerization (SIP) synthesized β-silicon carbide (β-SiC)/polyaniline (PANI) metacomposites. The calculated plasma frequency for these metacomposites with different loadings of β-SiC nanoparticles was ranging from 6.11 × 10(4) to 1.53 × 10(5) rad s(-1). The relationship between the negative permittivity and plasma frequency indicates the existence of switching frequency, at which the permittivity was changed from negative to positive. More interestingly, the synthesized non-magnetic metacomposites, observed to follow the 3-dimensional (3-D) Mott variable range hopping (VRH) electrical conduction mechanism, demonstrated high positive magnetoresistance (MR) values of up to 57.48% and high MR sensitivity at low magnetic field regimes.

  9. Physical mechanism determining the radial electric field and its radial structure in a toroidal plasma

    Energy Technology Data Exchange (ETDEWEB)

    Ida, Katsumi; Miura, Yukitoshi; Itoh, Sanae [and others

    1994-10-01

    Radial structures of plasma rotation and radial electric field are experimentally studied in tokamak, heliotron/torsatron and stellarator devices. The perpendicular and parallel viscosities are measured. The parallel viscosity, which is dominant in determining the toroidal velocity in heliotron/torsatron and stellarator devices, is found to be neoclassical. On the other hand, the perpendicular viscosity, which is dominant in dictating the toroidal rotation in tokamaks, is anomalous. Even without external momentum input, both a plasma rotation and a radial electric field exist in tokamaks and heliotrons/torsatrons. The observed profiles of the radial electric field do not agree with the theoretical prediction based on neoclassical transport. This is mainly due to the existence of anomalous perpendicular viscosity. The shear of the radial electric field improves particle and heat transport both in bulk and edge plasma regimes of tokamaks. (author) 95 refs.

  10. Determination of salicylic acid by HPLC in plasma and saliva from children with juvenile chronic arthritis.

    Science.gov (United States)

    Legaz, M E; Acitores, E; Valverde, F

    1992-12-01

    A high performance liquid chromatography (HPLC) method has been developed for measuring salicylic acid in the plasma and saliva of children with juvenile chronic arthritis (JCA). Samples were extracted with diethyl ether and, after drying, redissolved in methanol to be chromatographed. Quantitation of salicylic acid was performed by reverse phase HPLC on a spherisorb ODS-2 column, using methanol: water: acetic acid as mobile phase. Phenolic was monitored by absorbance at 237 nm. Linearity between the amount of mass injected and the response in the detector was determined. This method was applied to compare concentrations of salivary and plasma salicylic acid. The method also permitted the quantitation of salivary salicylate as a non-invasive, indirect method for monitoring the concentration of plasma salicylate in patients with JCA.

  11. Validated HPLC-MS-MS method for determination of azithromycin in human plasma.

    Science.gov (United States)

    Barrett, B; Borek-Dohalský, V; Fejt, P; Vaingátová, S; Huclová, J; Nemec, B; Jelínek, I

    2005-09-01

    A validated, highly sensitive, and selective HPLC method with MS-MS detection has been developed for quantitative determination of azithromycin (AZI) in human Na2EDTA plasma. Roxithromycin (ROX) was used as internal standard. Human plasma containing AZI and internal standard was ultrafiltered through Centrifree Micropartition devices and the concentration of AZI was determined by isocratic HPLC-MS-MS. Multiple reaction monitoring mode (MRM) was used for MS-MS detection. The calibration plot was linear in the concentration range 2.55-551.43 ng mL(-1). Inter-day and Intra-day precision and accuracy of the proposed method were characterized by R.S.D and percentage deviation, respectively; both were less than 8%. Limit of quantification was 2.55 ng mL(-1). The proposed method was used to determine the pharmacokinetic profile of AZI (250-mg tablets).

  12. High-performance liquid chromatographic determination of memantine hydrochloride in rat plasma using sensitive fluorometric derivatization.

    Science.gov (United States)

    Xie, Mei-Fen; Zhou, Wei; Tong, Xin-Yi; Chen, Yi-Le; Cai, Yi; Li, Yan; Duan, Geng-Li

    2011-02-01

    In this study, we investigated a simple, sensitive and reliable liquid chromatography-fluorescence detection method for the determination of memantine hydrochloride in rat plasma which was based on derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). For the first time, FMOC-Cl was introduced into derivatization of memantine hydrochloride in rat plasma. The amino groups of memantine hydrochloride and amantadine hydrochloride (internal standard) were trapped with FMOC-Cl to form memantine hydrochloride-FMOC-Cl and amantadine hydrochloride-FMOC-Cl compositions, which can be very compatible for LC-FLD. Precipitation of plasma proteins by acetonitrile was followed by vortex mixing and centrifugation. Chromatographic separation was performed on a C(18) column (DIAMONSIL 150 × 4.6 mm, id 5 μm) with a mobile phase consisting of acetonitrile and water at a flow rate of 1.0 mL/min. The retention times of memantine hydrochloride-FMOC-Cl and amantadine hydrochloride-FMOC-Cl compositions were 23.69 and 40.27 min, respectively. Optimal conditions for the derivatization of memantine hydrochloride were also described. The limit of quantification (LOQ) was 25 ng/mL for memantine hydrochloride in plasma, the linear range was 0.025-5.0 μg/mL in plasma with a correlation coefficient (r) of 0.9999. The relative standard deviations (RSDs) of intra-day and inter-day assays were 4.46-12.19 and 5.23-11.50%, respectively. The validated method was successfully applied to the determination of memantine hydrochloride in rat plasma samples.

  13. Determination of total plasma hydroperoxides using a diphenyl-1-pyrenylphosphine fluorescent probe.

    Science.gov (United States)

    Santas, Jonathan; Guardiola, Francesc; Rafecas, Magda; Bou, Ricard

    2013-03-01

    Plasma hydroperoxides (HPs) are widely accepted to be good indicators of oxidative stress. By means of the method proposed here, which uses diphenyl-1-pyrenylphosphine (DPPP) as a fluorescent probe, all types of plasma HP were determined. The limits of detection and quantification of the method were 0.08 and 0.25 nmol of cumene hydroperoxide (CHP) equivalents in 40 μl of plasma, respectively. The method is satisfactory in terms of precision (5.3% for 14.5 μM CHP eq., n=8), and the recoveries were 91% and 92% after standard additions of 26 and 52 μM CHP, respectively. The selectivity of the proposed method is higher than 96%. Moreover, optimization of the reaction conditions and the addition of ethylenediaminetetraacetic acid (EDTA) disodium salt and 2,6-di-tert-butyl-4-methylphenol (BHT) prevented the formation of HP artifacts during the analysis. Therefore, the proposed method is useful for simple and quantitative determination of total plasma HPs.

  14. Elevated progesterone during ovarian stimulation for IVF

    DEFF Research Database (Denmark)

    Al-Azemi, M; Kyrou, D; Kolibianakis, E M

    2012-01-01

    of Medline and PubMed were searched to identify relevant publications. Good-quality evidence supports the negative impact on endometrial receptivity of elevated progesterone concentrations at the end of the follicular phase in ovarian stimulation. Future trials should document the cause and origin...... phase in ovarian stimulation. The databases of Medline and PubMed were searched to identify relevant publications. Good-quality evidence supports the negative impact on endometrial receptivity of elevated progesterone concentrations at the end of follicular phase in ovarian stimulation. Future trials...

  15. Modulation of Progesterone Receptor Isoform Expression in Pregnant Human Myometrium

    Directory of Open Access Journals (Sweden)

    Marina Ilicic

    2017-01-01

    Full Text Available Background. Regulation of myometrial progesterone receptor (PR expression is an unresolved issue central to understanding the mechanism of functional progesterone withdrawal and initiation of labor in women. Objectives. To determine whether pregnant human myometrium undergoes culture-induced changes in PR isoform expression ex situ and, further, to determine if conditions approaching the in vivo environment stabilise PR isoform expression in culture. Methods. Term nonlaboring human myometrial tissues were cultured under specific conditions: serum supplementation, steroids, stretch, cAMP, PMA, PGF2α, NF-κB inhibitors, or TSA. Following 48 h culture, PR-T, PR-A, and PR-B mRNA levels were determined using qRT-PCR. PR-A/PR-B ratios were calculated. Results. PR-T and PR-A expression and the PR-A/PR-B ratio significantly increased in culture. Steroids prevented the culture-induced increase in PR-T and PR-A expression. Stretch blocked the effects of steroids on PR-T and PR-A expression. PMA further increased the PR-A/PR-B ratio, while TSA blocked culture-induced increases of PR-A expression and the PR-A/PR-B ratio. Conclusion. Human myometrial tissue in culture undergoes changes in PR gene expression consistent with transition toward a laboring phenotype. TSA maintained the nonlaboring PR isoform expression pattern. This suggests that preserving histone and/or nonhistone protein acetylation is critical for maintaining the progesterone dependent quiescent phenotype of human myometrium in culture.

  16. Modulation of Progesterone Receptor Isoform Expression in Pregnant Human Myometrium

    Science.gov (United States)

    2017-01-01

    Background. Regulation of myometrial progesterone receptor (PR) expression is an unresolved issue central to understanding the mechanism of functional progesterone withdrawal and initiation of labor in women. Objectives. To determine whether pregnant human myometrium undergoes culture-induced changes in PR isoform expression ex situ and, further, to determine if conditions approaching the in vivo environment stabilise PR isoform expression in culture. Methods. Term nonlaboring human myometrial tissues were cultured under specific conditions: serum supplementation, steroids, stretch, cAMP, PMA, PGF2α, NF-κB inhibitors, or TSA. Following 48 h culture, PR-T, PR-A, and PR-B mRNA levels were determined using qRT-PCR. PR-A/PR-B ratios were calculated. Results. PR-T and PR-A expression and the PR-A/PR-B ratio significantly increased in culture. Steroids prevented the culture-induced increase in PR-T and PR-A expression. Stretch blocked the effects of steroids on PR-T and PR-A expression. PMA further increased the PR-A/PR-B ratio, while TSA blocked culture-induced increases of PR-A expression and the PR-A/PR-B ratio. Conclusion. Human myometrial tissue in culture undergoes changes in PR gene expression consistent with transition toward a laboring phenotype. TSA maintained the nonlaboring PR isoform expression pattern. This suggests that preserving histone and/or nonhistone protein acetylation is critical for maintaining the progesterone dependent quiescent phenotype of human myometrium in culture. PMID:28540297

  17. Relationship between Insuline-like Growth Factor-Ⅰ and Progesterone Secretion of Cultured Human Trophoblast Cells in Vitro

    Institute of Scientific and Technical Information of China (English)

    Xiao-jin ZHANG; Sui-qi GUI; Lin CAO; Zu-yue SUN

    2007-01-01

    Objective To investigate the effect of insuline-like growth factor-Ⅰ(IGF-Ⅰ) on progesterone genesis and regulation.Methods Cytotrophoblast cells were collected by trypsin-collagenase digestion and percoll gradient centrifugation for primary culture.After stimulated with different concentrations(1 00 μg/ml,10 μg/ml,1μg/ml,0.1 μg/ml) of IGF-Ⅰat the same time and with different duration(12 h, 24 h, 48 h,72 h) of IGF-Ⅰ with the same concentration, progesterone levels in the media were measured by radioimmunoassay.Simultaneously,semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) was applied to determine the expression of low density lipoprotein receptor (LDLR) mRNA.Results Progesterone levels correlated positively with IGF-Ⅰalong with the IGF-Ⅰconcentration increasing,progesterone level began to increase at 12 h,and reached the climax at 48 h when cultured with 100 μg/L IGF-Ⅰ. The expression of LDLR mRNA was detectable in every group and accordant with variation of progesterone level.Conclusion Progesterone secretion has time- and dose-dependent effect on IGF-Ⅰ,and IGF-Ⅰcan up-regulate the expression of LDLR mRNA. IGF-Ⅰmay play an important role in promoting secretion of progesterone in trophoblast cells.

  18. Indirect determination of the electric field in plasma discharges using laser-induced fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Vaudolon, J., E-mail: julien.vaudolon@cnrs-orleans.fr; Mazouffre, S., E-mail: stephane.mazouffre@cnrs-orleans.fr [CNRS - ICARE (Institut de Combustion Aérothermique Réactivité et Environnement), 1 C Av. de la Recherche Scientifique, 45071 Orléans Cedex 2 (France)

    2014-09-15

    The evaluation of electric fields is of prime interest for the description of plasma characteristics. In this work, different methods for determining the electric field profile in low-pressure discharges using one- and two-dimensional Laser-Induced Fluorescence (LIF) measurements are presented and discussed. The energy conservation, fluid, and kinetic approaches appear to be well-suited for the electric field evaluation in this region of the plasma flow. However, the numerical complexity of a two-dimensional kinetic model is penalizing due to the limited signal-to-noise ratio that can be achieved, making the computation of the electric field subject to large error bars. The ionization contribution which appears in the fluid model makes it unattractive on an experimental viewpoint. The energy conservation and 1D1V kinetic approaches should therefore be preferred for the determination of the electric field when LIF data are used.

  19. Determination of catecholamines in plasma by HPLC and amperometric detection. Comparison with a radioenzymatic method.

    Science.gov (United States)

    Bauersfeld, W; Ratge, D; Knoll, E; Wisser, H

    1986-03-01

    The determination of norepinephrine and epinephrine in plasma by HPLC with amperometric detection was modified, giving detection limits of 25 ng/l for norepinephrine and epinephrine, respectively, using 1 ml plasma. In order to achieve this sensitivity, it was necessary to minimize the background noise by modification of instrumentation and specimen handling. Particularly important was the extra purification of the reagents, the application of micro-bore HPLC, the enzymatic cleavage of uric acid and temperature control of the amperometric cell and the amplifier. Comparison of the present method with the radioenzymatic determination of catecholamines resulted in coefficients of correlation of r = 0.924 and 0.919 for norepinephrine and epinephrine, resp. (n = 38). The concentrations of the 38 different samples used for the comparison were in the physiological range.

  20. Fetal sex determination of macaque monkeys by a nested PCR using maternal plasma.

    Science.gov (United States)

    Mitsunaga, Fusako; Ueiwa, Miyuki; Kamanaka, Yoshirou; Morimoto, Mayumi; Nakamura, Shin

    2010-01-01

    Non-invasive fetal sex determination is required for biomedical studies, in which some sexual difference would be expected in fetal events, in order to make a choice of male or female fetus. To detect male fetal DNA of the sex-determining region Y gene (SRY) in maternal macaque plasma, nested real-time PCR using the SYBR Green system was developed. In all cases of pregnant macaques with male fetuses, a nested PCR product of SRY was amplified from the mother's plasma, while no amplicon was detected in any case of pregnancy with a female fetus. Interestingly, fetal SRY DNA appeared to be cleared rapidly from the maternal blood after parturition. The current method is sensitive and can be performed with a regular PCR machine.

  1. Liquid chromatography-mass spectrometric determination of rufinamide in low volume plasma samples.

    Science.gov (United States)

    Gáll, Zsolt; Vancea, Szende; Dogaru, Maria T; Szilágyi, Tibor

    2013-12-01

    Quantification of rufinamide in plasma was achieved using a selective and sensitive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method. The chromatographic separation was achieved on a reversed phase column (Zorbax SB-C18 100mm×3mm, 3.5μm) under isocratic conditions. The mobile phase consisted of a mixture of water containing 0.1% formic acid and methanol (50:50, v/v). The mass spectrometric detection of the analyte was in multiple reaction monitoring mode (MRM) using an electrospray positive ionization (ESI positive). The monitored ions were 127m/z derived from 239m/z rufinamide and 108m/z derived from 251m/z the internal standard (lacosamide). Protein precipitation with methanol was applied for sample preparation using only 50μl aliquots. The concentration range was 40-2000ng/ml for rufinamide in plasma. The limit of detection was 1.25ng/ml and the lower limit of quantification was established at 5ng/ml rufinamide concentration. Selectivity and matrix effect was verified using individual human, rat and rabbit plasma samples. Short-term, post-preparative and freeze-thaw stability was also investigated. The proposed method provides accuracy, precision and high-throughput (short runtime 4.5min) for quantitative determination of rufinamide in plasma. This is the first reported liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for analysis of rufinamide from low volume plasma samples. The LC-MS/MS method was validated according to the current official guidelines and can be applied to accurately measure rufinamide level of large number of plasma samples from clinical studies or therapeutic drug monitoring.

  2. Spectrophotometric determination of total proteins in blood plasma: a comparative study among dye-binding methods

    OpenAIRE

    Dimas Augusto Morozin Zaia; Fábio Rangel Marques; Cássia Thaïs Bussamra Vieira Zaia

    2005-01-01

    A comparative study between the biuret method (standard method for total proteins) and spectrophotometric methods using dyes (Bradford, 3',3",5',5"-tetrabromophenolphthalein ethyl ester-TBPEE, and erythrosin-B) was carried out for the determination of total proteins in blood plasma from rats. Bradford method showed the highest sensitivity for proteins and biuret method showed the lowest. For all the methods, the absorbance for different proteins (BSA, casein, and egg albumin) was measured and...

  3. Determination of diclofenac concentrations in human plasma using a sensitive gas chromatography mass spectrometry method

    OpenAIRE

    2016-01-01

    Background A gas chromatography mass spectrometry (GCMS) method for the determination of diclofenac in human plasma has been developed and validated. Results This method utilizes hexane which is a relatively less toxic extraction solvent compared to heptane and benzene. In addition, phosphoric acid and acetone were added to the samples as deproteination agents, which increased the recovery of diclofenac. These revised processes allow clean extraction and near-quantitative recovery of analyte ...

  4. Determination of diclofenac concentrations in human plasma using a sensitive gas chromatography mass spectrometry method

    OpenAIRE

    Shah, Iltaf; Barker, James; Naughton, Declan P; Barton, Stephen J.; Ashraf, Syed Salman

    2016-01-01

    Background\\ud A gas chromatography mass spectrometry (GCMS) method for the determination of diclofenac in human plasma has been developed and validated.\\ud \\ud Results\\ud This method utilizes hexane which is a relatively less toxic extraction solvent compared to heptane and benzene. In addition, phosphoric acid and acetone were added to the samples as deproteination agents, which increased the recovery of diclofenac. These revised processes allow clean extraction and near-quantitative recover...

  5. Determination of Ovsynch Efficiency for Estrus Synchronization by Plasma LH and P4 Levels in Nili Ravi Buffalo during Peak and Low Breeding Seasons

    Directory of Open Access Journals (Sweden)

    S. Jabeen, M. Anwar1*, S. M. H. Andrabi1, A. Mehmood1, S. Murtaza2 and M. Shahab

    2013-04-01

    Full Text Available Oestrus synchronization may be used to overcome poor oestrus expression and seasonality of breeding in buffalo (Bubalus bubalis. The present work was conducted to compare the efficiency of an oestrus synchronization protocol “ovsynch” in Nili Ravi buffalo during peak (n=8 and low breeding seasons (n=11 by determining luteinizing hormone (LH peak and progesterone (P4 rise in blood plasma using ELISA. Buffaloes were administered gonadotropin releasing hormone (GnRH analogue (50 µg lecirelin; day 0 followed by prostaglandinF2α (PGF2α analogue (150 µg cloprostenol; day 7 and again GnRH analogue at 36 hours after PGF2α. Blood sampling for LH was started 12 h after PGF2α injection and done at 3 h interval up to 108 h. An animal was considered to have responded to ovsynch protocol if it showed LH peak within 48 h after PGF2α injection (and within 3-6 h after second GnRH injection and showed a P4 concentration of >2.0 ng/ml on day 18 after the 1st GnRH injection. 87.5% animals showed positive response to ovsynch protocol in term of oestrus and ovulation synchronization during the peak season as assessed by LH peak and P4 level while only 36.36 % animals responded to synchronization treatment during low breeding season (P<0.05. The LH peak in responsive animals was 11.48±1.98 and 10.38±1.77 ng/ml in peak and low breeding season respectively. It is concluded that ovsynch protocol in buffaloes resulted in a significantly better response during peak breeding season as compared to that in low breeding season.

  6. High Performance Liquid Chromatography Tandem Mass Spectrometry Assay for the Determination of Cobinamide in Pig Plasma

    Science.gov (United States)

    McCracken, Brent A.; Brittain, Matthew K.

    2015-01-01

    Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been widely utilized for the analysis of compounds in biological matrices due to its selectivity and sensitivity. This study describes the application of an LC-MS/MS-based approach toward the analysis of cobinamide in Yorkshire pig plasma. The selectivity, accuracy, precision, recovery, linearity, range, carryover, sensitivity, matrix effect, interference, stability, reproducibility, and ruggedness of the method were investigated in pig plasma. The accuracy and precision of the method was determined to be within 10% over three different days over a range of concentrations (25–10,000 ng/mL) that spanned more than two orders of magnitude. The lower limit of quantitation (LLOQ) for dicyanocobinamide was determined to be 25 ng/mL in pig plasma. Carryover was acceptable, as the area response of the carryover blanks were ≤15% of the area response of the nearest LLOQ standard for the analyte, while it was nonexistent for the internal standard. Specificity was ensured using six different lots of pig plasma. While the matrix effects of dicyanocobinamide in plasma were enhanced, ginsenoside Rb1 experienced signal suppression under the described conditions. The absolute recovery results for both compounds were consistent, precise, and reproducibly lower than expected at ~60% for dicyanocobinamide and ~22% for ginsenoside Rb1, confirming that a matrix standard curve was required for accurate quantitation. Cobinamide was shown to be very stable in matrix at various storage conditions including room temperature, refrigerated, and frozen at time intervals of 20 hours, 4 days, and 60 days respectively. This method was demonstrated to be sensitive, reproducible, stable, and rugged, and it should be applicable to the analysis of cobinamide in other biological matrices and species. PMID:26540437

  7. Toward a dry reagent immunoassay of progesterone in bovine milk

    NARCIS (Netherlands)

    Posthuma-Trumpie, Geertruida Afina

    2008-01-01

    This thesis is aimed at the development of a dry reagent immunoassay of progesterone in cow's milk. Progesterone is a steroid hormone and regulates ovulation in female mammals. The concentration of progesterone in blood and in milk is in accordance with the reproductive cycle of the individual femal

  8. PH metric method for the determination of nicotinic acid in plasma.

    Science.gov (United States)

    Gravesen, J

    1977-04-01

    The acidimetric method for the determination of nicotinic acid (NA) using Lactobacillus plantarum ATCC 8014 (Lactobacillus arabinosus 17-5) has been simplified and thus made less time consuming, and the sensitivity has been increased fivefold by replacement of the titration by a pH determination. As the regression of the decrease in pH on the amount of NA was found linear within a range of 1 to 4 ng of NA per ml, the calculations were performed according to the slope-ratio principle. The NA concentration of plasma was determined with a coefficient of variation of 5 to 7%, which rose to about 10% at low NA concentrations. Assays of fasting plasma samples from 13 hyperlipidemic male patients showed a group mean NA concentration of 80 +/- 55 ng/ml (mean +/- 2 standard deviation), before treatment, and 705 +/- 544 ng/ml (mean +/- 2 standard deviation) during therapy with sustained release NA preparations, of which a single dose, ingested during steady-state conditions, doubled or tripled the plasma concentration within 1 to 3 h.

  9. Influence of monoolein on progesterone transdermal delivery

    Directory of Open Access Journals (Sweden)

    Wanessa de Souza Cardoso Quintão

    2015-12-01

    Full Text Available abstract This work aimed to investigate in vitro the influence of monoolein (MO on progesterone (PG transdermal delivery and skin retention. Information about the role of MO as an absorption enhancer for lipophilic molecules can help on innovative product development capable of delivering the hormone through the skin in a consistent manner, improving transdermal therapy of hormonal replacement. MO was dispersed in propylene glycol under heat at concentrations of 0% (control, 5% w/w, 10% w/w and 20% w/w. Then, 0.6% of PG (w/w was added to each formulation. Permeation profile of the hormone was determined in vitro for 48 h using porcine skin in Franz diffusion cells. PG permeation doubled when 5% (w/w of MO was present in formulation in comparison to both the control and higher MO concentrations (10% and 20% w/w. An equal trend was observed for PG retention in stratum corneum (SC and reminiscent skin (E+D. PG release rates from the MO formulations, investigated using cellulose membranes, revealed that concentrations of MO higher than 5% (w/w hindered PG release, which indeed negatively reflected on the hormone permeation through the skin. In conclusion, this work demonstrated the feasibility of MO addition (at 5% w/w in formulations as a simple method to increase transdermal PG delivery for therapies of hormonal replacement. In contrast, higher MO concentrations (from 10% to 20% w/w can control active release, and this approach could be extrapolated to other lipophilic, low-molecular-weight molecules.

  10. Determination and pharmacokinetic study of pirfenidone in rat plasma by UPLC-MS/MS.

    Science.gov (United States)

    Sun, Wei; Jiang, Zhe-li; Zhou, Lei; Chen, Rui-min; Wang, Zhe; Li, Wan-shu; Jiang, Shuo-min; Hu, Guo-xin; Chen, Rui-jie

    2015-02-15

    A rapid, sensitive and selective ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed and validated for the determination and pharmacokinetic investigation of pirfenidone in rat plasma. Sample preparation was accomplished through a simple one-step deproteinization procedure with 0.2 mL of acetonitrile to a 0.1 mL plasma sample. Plasma samples were separated by UPLC on an Acquity UPLC BEH C18 column using a mobile phase consisting of acetonitrile-0.1% formic acid in water with gradient elution. The total run time was 3.0 min and the elution of pirfenidone was at 1.39 min. The detection was performed on a triple quadrupole tandem mass spectrometer in the multiple reaction-monitoring (MRM) mode using the respective transitions m/z 186.2→92.1 for pirfenidone and m/z 237.1→194.2 for carbamazepine (IS), respectively. The calibration curve was linear over the range of 5-2000 ng/mL with a lower limit of quantitation (LLOQ) of 5 ng/mL. Mean recovery of pirfenidone in plasma was in the range of 80.4-84.3%. Intra-day and inter-day precision were both pirfenidone in rats.

  11. [Simultaneous determination of glucocorticoids in bovine plasma by high-pressure liquid chromatography].

    Science.gov (United States)

    Sasaki, Y; Tsuiki, M; Higashimaki, T

    1982-09-20

    A method is described for the simultaneous determination of major glucocorticoids in bovine plasma, that is, Cortisol, Corticosterone and Cortisone, which includes separation by high-pressure liquid chromatography and measurement using a UV detector (254 nm). Chromatography on the Zorbax CN column is carried out with the eluent system, that is, n-hexane/ethanol (=87/13 v/v). First, Dexamethasone was selected as the best internal standard under the previously mentioned conditions. The calibration curves for the glucocorticoids were all clearly linear. The recoveries for each 5 ng of added Cortisol, Corticosterone and Cortisone were 96.7 +/- 3.2%, 100.4 +/- 2.3% and 96.1 +/- 2.2%, respectively. The reproducibility of Cortisol was good and its coefficient of variation of intraassay was small (2.7%), whereas such coefficient was not so in Corticosterone (7.7%) and Cortisone (7.3%). This appears to be due to low concentrations of Corticosterone and Cortisone in the plasma. Specificity of this method to each glucocorticoid was ascertained using plasma from adrenalectomized wethers. In addition, the correlation coefficient between measurements of Cortisol by radioimmunoassay and those by this method was 0.978. In conclusion, the method described in this paper appears to be satisfactory for the simultaneous quantification of plasma glucocorticoids.

  12. Determination of fatty acid profile in ram spermatozoa and seminal plasma.

    Science.gov (United States)

    Díaz, R; Torres, M A; Bravo, S; Sanchez, R; Sepúlveda, N

    2016-08-01

    Fatty acids are important in male reproductive function because they are associated with membrane fluidity, acrosome reaction, sperm motility and viability, but limited information exists about the fatty acid profile of ram semen. Our aim was to determine the fatty acid composition in ram spermatozoa and seminal plasma. Sixty ejaculates were obtained from three ram (20 ejaculates/ram) using artificial vagina. Ram spermatozoa (RS) and seminal plasma (SP) were separated using centrifugation, and the fatty acids were analysed by gas chromatography. Total lipids obtained in ram spermatozoa were 1.8% and 1.6% in seminal plasma. Saturated fatty acid (SFA) was proportionally major in SP (66.6%) that RS (49.9%). The highest proportions of SFA corresponded to C4:0 (RS = 16.3% and SP = 28.8%) and C16:0 (RS = 16.3% and PS = 20%). The most important unsaturated fatty acid (UFA) was docosahexaenoic acid (DHA), 44.9% in RS and 31.5% in SP. The profile of fatty acid and their proportions showed differences between spermatozoa and seminal plasma.

  13. Determination of Febuxostat in Human Plasma Using RP-LC-UV Method.

    Science.gov (United States)

    Younes, Kareem M; El-Kady, Ehab F; Elzanfaly, Eman S

    2016-07-01

    A simple and sensitive bioanalytical high-performance liquid chromatographic method with ultraviolet detection was developed and validated for the quantification of febuxostat (FEB) in human plasma. Ketoprofen was used as an internal standard. The analytes were extracted from human plasma samples by precipitation with acetonitrile. The reconstituted samples were chromatographed on C18 Bondapack 10 µm, 250 × 4.6 mm, Waters Column (Ireland) by using a mixture of acetonitrile and 0.5% aqueous phosphoric acid (pH 3) (52 : 48, v/v) as the mobile phase. The chromatographic separation was isocratically performed at room temperature at a flow rate of 1.0 mL/min with UV detection at 315 nm. The method exhibited a linear dynamic range over 0.05-5.00 µg/mL FEB in human plasma. The lower limit of quantification was 0.05 µg/mL. The results of the intra- and interday precision and accuracy studies were within the acceptable limits. The validated method was successfully applied for the determination of FEB in human plasma samples for application in bioequivalence studies.

  14. Serum testosterone and progesterone levels and ovarian activity as indicators for seasonal breeding in dromedary camels in Sudan

    Directory of Open Access Journals (Sweden)

    Babiker, E. A,

    2011-05-01

    Full Text Available The present work was done to investigate the effect of season on level of testosterone, progesterone hormone and ovarian activity in Arabian dromedary camels. Over a period of one year (July 2009–June 2010, jugular blood samples were collected monthly from 7 mature male camels and 12 females at late pregnancy to detect the levels of testosterone and progesterone hormones. A total of 900 ovarian follicles were measured in the slaughterhouses of Tamboul and Um-Elgura to define the effect of season on ovarian activity. The obtained results showed that plasma testosterone levels greatly varied among months of the year. It increased during July and August and decreased during the period from September to February. The level started to increase again during March and remained high until the end of the study in June. Plasma progesterone level was high during July, while it dropped at parturition during August. The level of progesterone remained low during the period from September to February, before it started to rise again in March and remained high until the end of the study in June. The rise of progesterone level in females coincided with the rise of testosterone in males. Ovarian activity was observed throughout the different seasons with a maximum activity during autumn. According to the hormonal findings and ovarian activity, there is a clear breeding season in Arabian dromedary camel in the Butana area, northeast of Sudan extending throughout summer (March – June and autumn (July–October.

  15. Effect of Efavirenz on Endogenous Progesterone Concentrations and Contraceptive Outcomes among Ugandan HIV Infected Women Coadministering Ethinylestradiol/Levonorgestrel.

    Science.gov (United States)

    Munkwase, Grant; Bisaso, Kuteesa R; Kakaire, Othman; Nanzigu, Sarah

    2017-01-01

    This study assessed the effect of efavirenz mid-dose plasma concentrations on mid-luteal endogenous progesterone concentrations and contraceptive outcomes among 49 HIV infected women coadministering ethinylestradiol/levonorgestrel, including 34 HIV positive women on Highly Active Antiretroviral Therapy (HAART) and 15 HAART naïve HIV infected women, purposively selected from Mulago Hospital, Uganda. A blood sample was collected once between days 20 and 22 of each woman's menstrual cycle for measuring endogenous progesterone and efavirenz concentrations by electrochemiluminescence technology and High Performance Liquid Chromatography (HPLC), respectively. Descriptive statistical analysis and correlation and logistic regression analysis were done using SPSS v.21 and R3.1. Efavirenz showed a weak positive linear relationship with endogenous progesterone at efavirenz concentrations below 12 μg/ml. Based on serum endogenous progesterone, the observed hormonal contraceptives failure rate (24.5%) was higher than expected (maximum 8%). A higher proportion of HIV positive women on efavirenz based HAART (26.5%) was at risk of contraceptive failure than their HIV infected HAART naïve counterparts (20%) though it was not statistically significant (p = 0.63). Efavirenz mid-dose plasma concentrations seem to have no significant effect on mid-luteal endogenous progesterone concentrations and contraceptive outcomes among HIV infected Ugandan women coadministering ethinylestradiol/levonorgestrel oral pills.

  16. Effect of Efavirenz on Endogenous Progesterone Concentrations and Contraceptive Outcomes among Ugandan HIV Infected Women Coadministering Ethinylestradiol/Levonorgestrel

    Directory of Open Access Journals (Sweden)

    Grant Munkwase

    2017-01-01

    Full Text Available This study assessed the effect of efavirenz mid-dose plasma concentrations on mid-luteal endogenous progesterone concentrations and contraceptive outcomes among 49 HIV infected women coadministering ethinylestradiol/levonorgestrel, including 34 HIV positive women on Highly Active Antiretroviral Therapy (HAART and 15 HAART naïve HIV infected women, purposively selected from Mulago Hospital, Uganda. A blood sample was collected once between days 20 and 22 of each woman’s menstrual cycle for measuring endogenous progesterone and efavirenz concentrations by electrochemiluminescence technology and High Performance Liquid Chromatography (HPLC, respectively. Descriptive statistical analysis and correlation and logistic regression analysis were done using SPSS v.21 and R3.1. Efavirenz showed a weak positive linear relationship with endogenous progesterone at efavirenz concentrations below 12 μg/ml. Based on serum endogenous progesterone, the observed hormonal contraceptives failure rate (24.5% was higher than expected (maximum 8%. A higher proportion of HIV positive women on efavirenz based HAART (26.5% was at risk of contraceptive failure than their HIV infected HAART naïve counterparts (20% though it was not statistically significant (p=0.63. Efavirenz mid-dose plasma concentrations seem to have no significant effect on mid-luteal endogenous progesterone concentrations and contraceptive outcomes among HIV infected Ugandan women coadministering ethinylestradiol/levonorgestrel oral pills.

  17. HPLC-UV determination of metformin in human plasma for application in pharmacokinetics and bioequivalence studies.

    Science.gov (United States)

    Porta, Valentina; Schramm, Simone Grigoleto; Kano, Eunice Kazue; Koono, Eunice Emiko; Armando, Yara Popst; Fukuda, Kazuo; Serra, Cristina Helena Dos Reis

    2008-01-07

    In this study, a simple, rapid and sensitive HPLC method with UV detection is described for determination of metformin in plasma samples from bioequivalence assays. Sample preparation was accomplished through protein precipitation with acetonitrile and chromatographic separation was performed on a reversed-phase phenyl column at 40 degrees C. Mobile phase consisted of a mixture of phosphate buffer and acetonitrile at flow rate of 1.0 ml/min. Wavelength was set at 236 nm. The method was applied to a bioequivalence study of two drug products containing metformin, and allowed determination of metformin at low concentrations with a higher throughput than previously described methods.

  18. Fetal male lineage determination by analysis of Y-chromosome STR haplotype in maternal plasma.

    Science.gov (United States)

    Barra, Gustavo Barcelos; Santa Rita, Ticiane Henriques; Chianca, Camilla Figueiredo; Velasco, Lara Francielle Ribeiro; de Sousa, Claudia Ferreira; Nery, Lídia Freire Abdalla; Costa, Sandra Santana Soares

    2015-03-01

    The aim of this study is to determine the fetus Y-STR haplotype in maternal plasma during pregnancy and estimate, non-invasively, if the alleged father and fetus belong to the same male lineage. The study enrolled couples with singleton pregnancies and known paternity. All participants signed informed consent and the local ethics committee approved the study. Peripheral blood was collected in EDTA tubes (mother) and in FTA paper (father). Maternal plasma DNA was extracted by using NucliSens EasyMAG. Fetal gender was determined by qPCR targeting DYS-14 in maternal plasma and it was also confirmed after the delivery. From all included volunteers, the first consecutive 20 mothers bearing male fetuses and 10 mothers bearing female fetuses were selected for the Y-STR analysis. The median gestational age was 12 weeks (range 12-36). All DNA samples were subjected to PCR amplification by PowerPlex Y23, ampFLSTR Yfiler, and two in-house multiplexes, which together accounts for 27 different Y-STR. The PCR products were detected with 3500 Genetic Analyzer and they were analyzed using GeneMapper-IDX. Fetuses' haplotypes (Yfiler format) were compared to other 5328 Brazilian haplotypes available on Y-chromosome haplotypes reference database (YHRD). As a result, between 22 and 27 loci were successfully amplified from maternal plasma in all 20 cases of male fetuses. None of the women bearing female fetuses had a falsely amplified Y-STR haplotype. The haplotype detected in maternal plasma completely matched the alleged father haplotype in 16 out of the 20 cases. Four cases showed single mismatches and they did not configure exclusions; 1 case showed a mutation in the DYS 458 locus due to the loss of one repeat unit and 3 cases showed one DYS 385I/II locus dropout. All mismatches were confirmed after the delivery. Seventeen fetuses' haplotypes were not found in YHRD and one of them had a mutation, which corresponded to the paternity probability of 99.9812% and 95.7028%, respectively

  19. Effects of estradiol and progesterone on the variability of the micronucleus assay

    Energy Technology Data Exchange (ETDEWEB)

    Baeyens, Ans [Department of Anatomy, Embryology, Histology and Medical Physics, Ghent University, L. Pasteurlaan 2, 9000 Gent (Belgium)]. E-mail: ansbaeyens@hotmail.com; Vandersickel, Veerle [Department of Anatomy, Embryology, Histology and Medical Physics, Ghent University, L. Pasteurlaan 2, 9000 Gent (Belgium); Thierens, Hubert [Department of Anatomy, Embryology, Histology and Medical Physics, Ghent University, L. Pasteurlaan 2, 9000 Gent (Belgium); Ridder, Leo De [Department of Anatomy, Embryology, Histology and Medical Physics, Ghent University, L. Pasteurlaan 2, 9000 Gent (Belgium); Vral, Anne [Department of Anatomy, Embryology, Histology and Medical Physics, Ghent University, L. Pasteurlaan 2, 9000 Gent (Belgium)

    2005-10-15

    To investigate chromosomal radiosensitivity of lymphocytes the micronucleus (MN) assay has been used for many years. The results of these studies suggest the use of the MN assay as a biomarker for cancer predisposition. However, the MN assay has still some limitations associated with the reproducibility and sensitivity. Especially a high intra-individual variability has been observed. An explanation for this high intra-individual variability is not yet available. In literature it is suggested that the high variability among females is attributable to hormonal status. In this study we investigated if the high intra-individual variability in micronucleus formation in lymphocytes of females after in vitro exposure to ionising radiation is caused by variations in hormone levels of estradiol (E2) and progesterone (PROG). For this, the MN assay was performed on blood samples of 18 healthy women during 7 consecutive weeks while the estradiol and progesterone levels were determined at the same time. The MN assay was also examined in cultures of isolated blood lymphocytes with estradiol or progesterone levels added in vitro. The results demonstrated that estradiol and progesterone levels have no influence on the variations in radiation-induced MN yields observed in blood samples of healthy women. These conclusions were confirmed by the 'in vitro' experiments as no correlation between the MN yields and the concentrations of hormones (estradiol or progesterone) added in vitro to isolated lymphocytes cultures was observed.

  20. Epithelial progesterone receptor exhibits pleiotropic roles in uterine development and function.

    Science.gov (United States)

    Franco, Heather L; Rubel, Cory A; Large, Michael J; Wetendorf, Margeaux; Fernandez-Valdivia, Rodrigo; Jeong, Jae-Wook; Spencer, Thomas E; Behringer, Richard R; Lydon, John P; Demayo, Francesco J

    2012-03-01

    The ovarian steroid progesterone, acting through the progesterone receptor (PR), coordinates endometrial epithelial-stromal cell communication, which is critical for its development and function. PR expression in these cellular compartments is under tight temporal and endocrine control. Although ex vivo studies demonstrated the importance of stromal PR expression, they failed to show a role for epithelial PR in uterine function. Here, the in vivo role of PR in the uterine epithelium is defined using floxed PR (PR(f/f)) mice crossed to Wnt7a-Cre mice. Progesterone was unable to stimulate the expression of its epithelial target genes, including Ihh, in the Wnt7a-Cre(+)PR(f/-) mice. Analysis was conducted on Ihh to determine whether PR directly regulates epithelial gene transcription. ChIP-on-chip analysis identified PR binding sites in the 5'-flanking region of Ihh. Cotransfection of the proximal Ihh promoter with PR demonstrated that PR directly regulates Ihh transcription. Female Wnt7a-Cre(+)PR(f/-) mice are infertile due to defects in embryo attachment, stromal cell decidualization, and the inability to cease estrogen-induced epithelial cell proliferation. Finally, progesterone was unable to inhibit neonatal endometrial glandular development in Wnt7a-Cre(+)PR(f/-) mice. Thus, epithelial PR is necessary for the regulation of progesterone epithelial target gene expression, as well as uterine function and development.

  1. Progesterone treatment shows benefit in a pediatric model of moderate to severe bilateral brain injury.

    Directory of Open Access Journals (Sweden)

    Rastafa I Geddes

    Full Text Available PURPOSE: Controlled cortical impact (CCI models in adult and aged Sprague-Dawley (SD rats have been used extensively to study medial prefrontal cortex (mPFC injury and the effects of post-injury progesterone treatment, but the hormone's effects after traumatic brain injury (TBI in juvenile animals have not been determined. In the present proof-of-concept study we investigated whether progesterone had neuroprotective effects in a pediatric model of moderate to severe bilateral brain injury. METHODS: Twenty-eight-day old (PND 28 male Sprague Dawley rats received sham (n = 24 or CCI (n = 47 injury and were given progesterone (4, 8, or 16 mg/kg per 100 g body weight or vehicle injections on post-injury days (PID 1-7, subjected to behavioral testing from PID 9-27, and analyzed for lesion size at PID 28. RESULTS: The 8 and 16 mg/kg doses of progesterone were observed to be most beneficial in reducing the effect of CCI on lesion size and behavior in PND 28 male SD rats. CONCLUSION: Our findings suggest that a midline CCI injury to the frontal cortex will reliably produce a moderate TBI comparable to what is seen in the adult male rat and that progesterone can ameliorate the injury-induced deficits.

  2. Determination of nimodipine in plasma by HPLC-MS/MS and pharmacokinetic application

    Directory of Open Access Journals (Sweden)

    Demétrius Fernandes do Nascimento

    2010-12-01

    Full Text Available To develop and validate a rapid, specific and highly sensitive method to quantify nimodipine in human plasma using dibucaine as the internal standard (IS. The analyte and IS were extracted from plasma samples by liquid-liquid extraction using hexane-ethyl acetate (1:1 v/v. The chromatographic separation was performed on a Varian® Polaris C18 analytical column (3 μm, 50 x 2.0 mm and pre-column SecurityguardTM C18 (4.0 x 3.0 mm with a mobile phase of Acetonitrile-Ammonium acetate 0.02 ml/L (80:20v/v. The method had a chromatographic run time of 4.5 min and linear calibration curve over the range of 0.1- 40 ng/mL (r > 0.9938. The limit of quantification was 100 pg/mL. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. This validated method was successfully applied in determining the pharmacokinetic profile of nimodipine tablets of 30 mg administered to 24 healthy volunteers. The proposed method of analysis provided a sensitive and specific assay for nimodipine determination in human plasma. The time for the determination of one plasma sample was 4.5 min. This method is suitable for the analysis of nimodipine in human plasma samples collected for pharmacokinetic, bioavailability or bioequivalence studies in humans.Um método rápido, específico e sensível para quantificar nimodipino em plasma humano usando dibucaína como padrão interno (IS é descrito. O analito e o IS foram extraídos das amostras de plasma por extração líquido-líquido usando hexano-acetato de etila (1:1 v/v. A separação cromatográfica foi realizada utilizando-se uma coluna analítica C18 Varian® Polaris (3 μm, 50 x 2,0 mm e uma pré-coluna SecurityguardTM C18 (4,0 x 3,0 mm e acetonitrila-acetato de amônia 0,02 mol/L (80:20 v/v como fase móvel. O método apresentou tempo total de corrida de 4,5 min e curva de calibração linear com concentrações entre 0,1-40 ng/mL (r > 0,9938. O limite de quantificação foi de 100

  3. Determination of linsidomine in human plasma by tandem LC-MS with ESI.

    Science.gov (United States)

    Sutherland, F C; de Jager, A D; Swart, K J; Hundt, H K; Scanes, T; Hundt, A F

    2000-04-01

    A sensitive method for the determination of linsidomine in plasma was developed, using high-performance liquid chromatographic (HPLC) separation with tandem mass spectrometric detection. Linsidomine was derivatised with propyl chloroformate and extracted with tert-butyl methyl ether/1,2-dichloroethane (55:45, v/v), back-extracted into HCl (0.01 M) followed by alkalinisation and back-extraction into ether; the final ether extract evaporated, reconstituted in mobile phase and then separated on a Phenomenex Luna C18 (2) 5 micron 2.1 x 150 mm column with a mobile phase consisting of methanol water formic acid (98/100%) (400:600:0.05, v/v/v) at a flow-rate of 0.4 ml min(-1). Detection was achieved by a Finnigan MAT mass spectrometer (LCQ) at unit resolution in the selected reaction monitoring (SRM) mode monitoring the transition of the protonated molecular ion m/z 257.0 to the product ion m/z 86.0. The mean recovery for linsidomine was 51% with a lower limit of quantification of 0.70 ng/ml using 1 ml plasma for extraction. This LC-MS/MS method for the determination of linsidomine in human plasma allows for better specificity and a higher sample throughput than the traditional LC-UV methods. It also demonstrates the profound effect that the composition of acidic modifiers and matrix constituents can have on the electrospray ionisation (ESI) of the analyte.

  4. The determination of phenazone in blood plasma for obtained sistem suitable test of monitoring drug level

    Directory of Open Access Journals (Sweden)

    Mochamad Lazuardi

    2007-09-01

    Full Text Available The determining of Phenazone to human blood plasma from healthy man after separated by solid phase extraction (SPE and spectroscopic measurements has been investigated. The objective of that research was to obtain system suitable test for determine the Phenazone level in biological fluids (human blood plasma, for new performed dosage regimented in clinical dentistry. The method can be divided into the following four steps. 1. Centrifugation the blood sample, 2. Extraction from blood plasma and, 3. Separation by SPE with manual pressured, 4. Elution to SPE followed by the measurement on a spectrophotometer in the ultra violet region. The critical value of  │t │at the 5% confidence level indicates that there is no systematic error in the linearity proposed method. Recoveries for this research were obtained at ranging 93.460 to 95.598%. The coefficient variation precision of this procedure was clearly good at smallest than 2%. The analytical procedure can be carried out in one working operation as a monitored therapeutic activity.

  5. Determination of doxepin and desmethyldoxepin in human plasma using liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Badenhorst, D; Sutherland, F C; de Jager, A D; Scanes, T; Hundt, H K; Swart, K J; Hundt, A F

    2000-05-26

    A sensitive method for the simultaneous determination of doxepin and its active metabolite desmethyldoxepin in plasma was established, using high-performance liquid chromatographic separation with tandem mass spectrometric detection. The samples were extracted with hexane-isoamyl alcohol, separated on a Phenomenex Luna C18 5 microm, 150x2.1 mm column with a mobile phase consisting of methanol-water-formic acid (600:400:0.5, v/v) at a flow-rate of 0.25 ml/min. Detection was achieved by a Perkin-Elmer API 2000 mass spectrometer at unit resolution in multiple reaction monitoring mode monitoring the transition of the protonated molecular ions m/z 280.2, 266.2 and 250.1 to the product ions m/z 107.1, 107.1 and 191.0 for analyte, metabolite and internal standard (benzoctamine-HCl), respectively. TurbolonSpray ionisation was used for ion production. The mean recovery for doxepin and desmethyldoxepin was 90% and 75%, respectively, with a lower limit of quantification at 0.320 ng/ml and 0.178 ng/ml for the analyte and its metabolite, respectively, using 0.5 ml plasma for extraction. This is the first assay method described for the simultaneous determination of doxepin and desmethyldoxepin in plasma using LC-MS-MS. The method is sensitive enough to be used in drug bioavailability studies with doxepin.

  6. The determination of bromazepam in plasma by reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Le Solleu, H; Demotes-Mainard, F; Vinçon, G; Bannwarth, B

    1993-08-01

    A reversed-phase high-performance liquid chromatographic method has been developed for the determination of bromazepam, an anxiolytic benzodiazepine, in plasma. After a single-step extraction from alkalinized plasma with diethyl-ether in the presence of an internal standard (alpha-hydroxy-triazolam), the residues were chromatographed on a reversed-phase Nova Pak 5 microns C18 column, with a mobile phase of acetonitrile-water-triethylamine (700:300:4, v/v/v) adjusted to pH 7.4 with orthophosphoric acid. The limit of detection was 50 ng ml-1, using a 20 microliters injection with UV detection at 240 nm. Between-day and within-day relative standard deviations were lower than 6%. Studies of drug stability during sample storage at -20 degrees C and at +4 degrees C showed no degradation of bromazepam. However, bromazepam seemed to be degraded at ambient temperature, without any influence of light. This method is applied to the determination of bromazepam plasma levels in analytical toxicology.

  7. Cattle fetal sex determination by polymerase chain reaction using DNA isolated from maternal plasma.

    Science.gov (United States)

    da Cruz, A S; Silva, D C; Costa, E O A; De M-Jr, P; da Silva, C C; Silva, D M; da Cruz, A D

    2012-03-01

    The objective of this study was to evaluate the use of polymerase chain reaction analysis (PCR) of fetal cells/DNA in the maternal plasma of pregnant cows to determine the sex of the fetus. Plasma was harvested from 35 cows of mixed genotype at different stages of pregnancy ranging from 5 to 35 weeks. A male calf and a heifer calf provided the control samples. Fetal sex was determined by amplification of Y-specific sequences. For the 35 cows, the fetal sex predicted by this technique was in accordance with the sex of the calf at birth in 88.6% of cases. The agreement between predicted and observed fetal sex was less for cows with a gestational length of 35-48 days (63.6%). Regression analysis showed that there was a strong relationship between the probability of correctly predicting fetal sex and the stage of gestation. It was estimated that the test performed at 43.8 days post fertilization would have 95% accuracy, increasing to 99% accuracy for testing at 48.4 days and 99.9% accuracy for tests at 55.0 days or later. It was concluded that PCR analysis of fetal cells in maternal plasma can be used to predict successfully the sex of the fetus in cattle.

  8. Determination of Candesartan in Human Plasma with Liquid Chromatography - Tandem Mass Spectrometry.

    Science.gov (United States)

    Forjan, Vanja; Cvitkovič Maričič, Lea; Prosen, Helena; Brodnjak Vončina, Darinka

    2016-01-01

    A sensitive, specific and rapid liquid chromatography - tandem mass spectrometry method was developed and validated for the determination of candesartan in human plasma. Analyte was separated from endogenous components present in plasma by solid phase extraction. Chromatographic separation was performed on Gemini C18 analytical column using mobile phase acetonitrile - 5 mM ammonium formate pH 2 (90:10, v/v) at flow rate of 0.3 mL/min. For detection, tandem mass spectrometry in SRM mode with positive electrospray ionization was used. The mass transitions m/z 441.1 > 263.1 and 445.1 > 267.1 were used to determine candesartan by using candesartan-d4 as an internal standard. After development, the method was validated according to the requirements of EMA regulatory guidelines in the concentration range 1 - 400 ng/ml in human plasma. Limit of quantification (LLOQ) was 1 ng/ml. The developed and validated method proved to be very fast and reproducible and was therefore successfully implemented in pharmacokinetic and bioequivalence studies with large number of study samples.

  9. Determination of Alprazolam in Rabbit Plasma by GC-MS Method

    Directory of Open Access Journals (Sweden)

    Bilal Yilmaz

    2010-02-01

    Full Text Available This paper describes a gas chromatography-mass spectrometry (GC-MS method for determination of alprazolam in rabbit plasma. Alprazolam and internal standard (IS medazepam were extracted from plasma by using liquid-liquid extraction method. The samples were separated by GC on a DB-5MS analytical column and determined by a quadrupole mass spectrometer detector operated under selected ion monitoring mode (SIM. Excellent linearity was found between 50 and 1000 ng/mL r=0.998 for plasma samples. Intra-day and inter-day precisions expressed as the relative standard deviation (RSD for the method were 1.07-2.69% and 2.42-3.98%, respectively. The mean recovery of alprazolam samples was 98.82%. The limits of detection and uantification of alprazolam were 15 and 50 ng/mL, respectively. Also, the method was successfully applied to three New Zealand white abbits which had been given an oral tablet of 1.0 mg alprazolam.

  10. Kinetic and Related Determinants of Plasma Triglyceride Concentration in Abdominal Obesity: Multicenter Tracer Kinetic Study.

    Science.gov (United States)

    Borén, Jan; Watts, Gerald F; Adiels, Martin; Söderlund, Sanni; Chan, Dick C; Hakkarainen, Antti; Lundbom, Nina; Matikainen, Niina; Kahri, Juhani; Vergès, Bruno; Barrett, P Hugh R; Taskinen, Marja-Riitta

    2015-10-01

    Patients with obesity and diabetes mellitus have increased risk of cardiovascular disease. A major cause is an atherogenic dyslipidemia related primarily to elevated plasma concentrations of triglyceride-rich lipoproteins. The aim of this study was to clarify determinants of plasma triglyceride concentration. We focused on factors that predict the kinetics of very-low density lipoprotein 1 (VLDL1) triglycerides. A multicenter study using dual stable isotopes (deuterated leucine and glycerol) and multicompartmental modeling was performed to elucidate the kinetics of triglycerides and apoB in VLDL1 in 46 subjects with abdominal obesity and additional cardiometabolic risk factors. Results showed that plasma triglyceride concentrations were dependent on both the secretion rate (r=0.44, Ptriglycerides and VLDL1-apoB. Liver fat mass was independently and directly associated with secretion rates of VLDL1-triglycerides (r=0.56, Ptriglycerides (r=0.48, Ptriglyceride concentrations in abdominal obesity are determined by the kinetics of VLDL1 subspecies, catabolism being mainly dependent on apoC-III concentration and secretion on liver fat content. Reduction in liver fat and targeting apoC-III may be an effective approach for correcting triglyceride metabolism atherogenic dyslipidemia in obesity. © 2015 American Heart Association, Inc.

  11. Determination of trace amounts of morphine in human plasma by anodic adsorptive stripping differential pulse voltammetry

    Institute of Scientific and Technical Information of China (English)

    Ali Niazi; Ateesa Yazdanipour

    2008-01-01

    New adsorptive anodic differential pulse stripping voltammetry method for the direct determination of morphine at trace levels in human plasma of addicts is proposed.The procedure involves an adsorptive accumulation of morphine on a HMDE,followed by oxidation of adsorbed morphine by voltammetry scan using differential pulse modulation.The optimum conditions for the analysis of morphine are pH 10.5,Eacc of - 100 mV (vs.Ag/AgCl),and tacc of 120 s.The peak current is proportional to the concentration of morphine,and a linear calibration graph is obtained at 0.01-3.10 μg mL-1.A relative standard deviation of 1.06% (n=5) was obtained,and the limit of detection was 3 ng mL- 1.The capability of the method for the analysis of real samples was evaluated by the determination of morphine in spiked human plasma and addicts human plasma with satisfactory results.

  12. High-performance liquid chromatography determination of dipotassium clorazepate and its major metabolite nordiazepam in plasma.

    Science.gov (United States)

    Colin, P; Sirois, G; Lelorier, J

    1983-04-08

    A rapid and sensitive high-performance liquid chromatographic method is described for the quantitative analysis of dipotassium clorazepate (CZP) and its major metabolite nordiazepam (ND) in fresh human and dog plasma. The method consists of two separate selective ND extractions from a plasma sample without and with conversion of all the CZP to ND. For quantitation, diazepam (DZP) is used as the internal standard. The chromatographic phase utilized in a reversed-phase Hibar EC-RT analytical column prepacked with LiChrosolv RP-18 with a solvent system consisting of acetonitrile-0.05 M sodium acetate buffer, pH 5.0 (45:55). The UV absorbance is monitored at 225 nm using a variable-wavelength detector. The mean assay coefficient of variation over a concentration range of 20-400 ng per ml of plasma is less than 3% for the within-day precision. Recoveries of ND, DZP and CZP (as ND) are essentially quantitative at all levels investigated. The calibration curves of ND are rectilinear (r2 = 0.99) from the lower limit of sensitivity (2 ng/ml) to at least 2000 ng/ml in plasma. Applicability of the method to CZP and ND disposition studies in the anaesthetized mongrel dog is illustrated. When the two separate selective nordiazepam extractions from plasma cannot be performed immediately after blood sampling, an extrapolation kinetic method is suggested for the estimation of CZP concentration. In all previous in vivo studies, CZP has been determined only with gas-liquid chromatographic methods.

  13. Derivation and Testing of Computer Algorithms for Automatic Real-Time Determination of Space Vehicle Potentials in Various Plasma Environments

    Science.gov (United States)

    1988-05-31

    COMPUTER ALGORITHMS FOR AUTOMATIC REAL-TIME DETERMINATION OF SPACE VEHICLE POTENTIALS IN VARIOUS PLASMA ENVIRONMENTS May 31, 1988 Stanley L. Spiegel...crrnaion DiviSiofl 838 12 2 DERIVATION AND TESTING OF COMPUTER ALGORITHMS FOR AUTOMATIC REAL-TIME DETERMINATION OF SPACE VEHICLE POTENTIALS IN VARIOUS...S.L., "Derivation and testing of computer algorithms for automatic real time determination of space vehicle poteuatials in various plasma

  14. Progesterone and oestrogen in pregnancy and parturition: comparative aspects and hierarchical control.

    Science.gov (United States)

    Heap, R B; Galil, A K; Harrison, F A; Jenkin, G; Perry, J S

    1977-01-01

    The different ways in which the progesterone requirements of pregnancy are met in various species are reviewed here. Progesterone production expressed in terms of metabolic body weight lies within about one order of magnitude in several species (but not in the rat), whether the hormone is predominantly ovarian or placental in origin. Parturition is usually preceded by a decrease in the plasma concentration of progesterone and the evidence which suggests that a decrease in secretion involves enzyme induction is summarized. In the sheep a sequence of well-marked hormonal changes can be discerned--an increase in fetal cortisol secretion followed by a fall in placental progesterone and a rise in oestrogen and prostaglandin F2alpha secretion. This sequence has been interpreted in terms of hierarchical control theory which envisages a series of levels placed in a descending order of importance and in which there is a downward transmission of a signal from a higher level that modifies activity at a lower level. A dominant level in the hierarchy in the sheep and pig seems to be the fetal hypothalamus and pituitary, but in the rabbit the maternal hypothalamus, pituitary and ovary play a more dominant role.

  15. Evaluation of Serum Testosterone, Progesterone, Seminal Antisperm Antibody, and Fructose Levels among Jordanian Males with a History of Infertility

    Directory of Open Access Journals (Sweden)

    Hala I. Al-Daghistani

    2010-01-01

    Full Text Available Due to the biochemical complexity of seminal fluid, we attempt to study the possible correlation between fructose, which is secreted under the effect of androgen hormone, and autoimmunity, which might play a role in varicocele associated infertility, in reducing sperm motility. Seminal fructose, antisperm antibodies (ASAs and blood steroids hormones (testosterone and progesterone levels were measured in 66 infertile males with varicocele and 84 without varicocele referred for fertility treatment. Seminal analysis was performed with biochemical measurements of seminal fructose and mixed agglutination reaction (MAR for ASA. Serum levels of progesterone and testosterone were estimated using a competitive chemoluminescent enzyme immunoassay. The mean values for serum testosterone were 380.74±24.331, 365.9±16.55, and 367.5±21.8 ng/dl, progesterone 0.325±0.243, 0.341±0.022, and 0.357  ±  0.0306 ng/ml, and seminal plasma fructose 359.6  ±  26.75, 315.6  ±  13.08, and 332.08  ±  24.38 mg/dl in males with varicocele, without varicocele, and fertile males, respectively. A significant high level of testosterone was observed within varicocele group (P=.001. This result showed that testosterone may play a role as an infertility determinant in subjects with varicocele. ASA was detected in 18 (26.47% of cases with varicocele, 20 (38.46% without varicocele, and in 16 (32.0% fertile men. Cases with ASAs associated with low sperm motility morphology. An inverse correlation between sperm-bound antibodies and viscosity has been shown (P=.017. ASA showed some significant inverse relations with ages, durations of infertility, and viscosity (P<.05. In addition, a significant correlation was observed between ASA positive seminal plasma and testosterone concentration among infertile cases (with or without varicocele and fertile (P<.05. Our results suggest a relationship between testicular steroid hormone levels with

  16. Determinants of plasma pepsinogen levels in a population at high risk for stomach cancer in Venezuela.

    Science.gov (United States)

    Kato, I; Miki, K; Muñoz, N; Vivas, J H; Lopez, G; Peraza, S; Carillo, E; Castro, D; Andrade, O; Sanchez, V

    1995-09-04

    Determinants of plasma pepsinogens (PG) levels were studied in 1365 participants in a chemoprevention trial for gastric pre-cancerous lesions being conducted in Venezuela. Gastric biopsies, plasma samples and information on smoking and dietary habits were obtained at baseline examination. Both PG-I and PG-II levels increased progressively with the level of Helicobacter pylori infection in gastric biopsies, resulting in no clear trend in the I/II ratio. Instead, there was a progressive decrease in the I/II ratio with increasing degrees of infiltration of polynuclear cells and monocytes, atrophy, intestinal metaplasia and the stage of pre-cancerous lesions. The mean I/II ratios for atrophic gastritis or more advanced lesions were less than 4.0. When subjects with the I/II ratio 4 or higher were used as controls, severe reduction in the I/II ratio (food on the development of atrophic gastritis need to be studied further.

  17. High performance liquid chromatography method for rapid and accurate determination of homocysteine in plasma and serum

    DEFF Research Database (Denmark)

    Vester, Birte; Rasmussen, K

    1991-01-01

    Determination of homocysteine in plasma or serum for evaluation of cobalamin and folate deficiency is becoming an important diagnostic procedure. Accurate, rapid and low cost methods for measuring homocysteine are therefore required. We have improved an HPLC method and made it suitable for clinical...... application. The more important changes are the addition of an internal standard, mercaptopropionylglycine, and the use of a plasma/serum based calibration material. The method consists of the following steps: reduction of the sample with tri-n-butylphosphine, precipitation of proteins, derivatisation...... with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate, and HPLC separation followed by fluorescence detection. The linearity of the assays is established and the coefficient of variation is 3.0%. Stability studies show that blood samples must be cooled or centrifuged immediately after venipuncture...

  18. Single-dose subcutaneous iodine-131-iodohippurate for determination of renal plasma flow

    Energy Technology Data Exchange (ETDEWEB)

    Grant, M.E.; Herron, K.G.; MacDougall, M.L.; Preston, D.F.; Moore, W.V.; Wiegmann, T.B. (Department of Medicine, University of Kansas Medical Center, Kansas City (USA))

    1991-07-01

    Subcutaneous administration of a single dose of 131I-iodohippurate was used for determination of renal plasma flow (RPF) in 20 subjects during water diuresis. Slow release of tracer (200 microCi) permitted serial clearance measurements over 5 hr that were compared to standard, constant infusion, PAH clearance (mean 379.5 {plus minus} 34.9 ml/min/1.73 m2, range 50.9 to 696.3 ml/min/1.73 m2). RPF(Isotope) was 424.9 {plus minus} 30.3 ml/min/1.73 m2 (range 144.4 to 746.5 ml/min/1.73 m2) and highly correlated with RPFPAH (r = 0.883, p less than 0.0001). This technique permits prolonged studies of renal plasma flow under steady-state conditions without constant infusion.

  19. A comparison among optical emission spectroscopic methods of determining electron temperature in low pressure argon plasmas

    Institute of Scientific and Technical Information of China (English)

    Niu Tian-Ye; Cao Jin-Xiang; Liu Lei; Liu Jin-Ying; Wang Yan; Wang Liang

    2007-01-01

    In this article, four kinds of optical emission spectroscopic methods of determining electron temperature are used to investigate the relationship between electron temperature and pressure in the cylindrical plasmas of dc glow discharges at low pressures in laboratory by measuring the relative intensities of ArI lines at various pressures. These methods are developed respectively on the basis of the Fermi-Dirac model, corona model, and two kinds of electron collision cross section models according to the kinetic analysis. Their theoretical bases and the conditions to which they are applicable are reviewed, and their calculation results and fitting errors are compared with each other. The investigation has indicated that the electron temperatures obtained by the four methods become consistent with each other when the pressure increases in the low pressure argon plasmas.

  20. Determination of barnidipine in human serum and dog plasma by HPLC with electrochemical detection.

    Science.gov (United States)

    Takamura, K; Abdel-Wadood, H M; Kusu, F; Rafaat, I H; Saleh, G A; el-Rabbat, N A; Otagiri, M

    1995-10-01

    Barnidipine is a 1,4-dihydropyridine calcium antagonist. HPLC was conducted on a polybutadiene coated alumina column using an alkaline mobile phase and an electrochemical detector to determine the content of this drug in serum and plasma. A good linear relationship between barnidipine concentration and peak height was found in 5-500 ng/ml with a correlation coefficient of 0.998. The detection limit was 1 ng/ml. The within-day and day-to-day variations were examined for control human serum. Relative standard deviation of within-day assay for serum spiked with 10 ng/ml barnidipine.HCl was 6.9% and the recovery was 104%. A pharmacokinetic study was made in which the time course of barnidipine in dog plasma was followed.

  1. Development of a chromatographic method for the determination of saquinavir in plasma samples of HIV patients.

    Science.gov (United States)

    Campanero, M A; Escolar, M; Arangoa, M A; Sádaba, B; Azanza, J R

    2002-02-01

    A simple, sensitive and reproducible high-performance liquid chromatographic method for detecting and quantifying saquinavir in human plasma is described. Verapamil was used as internal standard. The method employes a single liquid-liquid extraction step with tert-butil methyl ether followed by chromatography on a Lichrospher 60 Select B C8 reversed-phase column. Ultraviolet detection was used to identify the compounds of interest. The quantitation limit of saquinavir was 1 ng/mL and only 0.5 mL of plasma sample was required for the determination. The average saquinavir recoveries over a concentration range of 2.5-500 ng/mL ranged from 86 to 95%. Precision and accuracy did not exceed 5%.

  2. GLC determination of plasma drug levels after oral administration of clorazepate potassium salts.

    Science.gov (United States)

    Hoffman, D J; Chun, A H

    1975-10-01

    Plasma nordiazepam levels resulting from the oral administration of clorazepate potassium salts were determined by a sensitive GLC assay. Nordiazepam and the internal standard (diazepam) were selectively extracted into ether at pH 9.2, hydrolyzed to their respective benzophenones, and quantified by electron-capture detection. The assay was used in a comparative bioavailability study of single equimolar oral doses of monopotassium and dipotassium salts of clorazepate in dogs. Both clorazepate salts were rapidly absorbed and exhibited mean peak total drug levels after 1 hr. Clorazepate levels accounted for about 50% of the total drug levels present. No statistical difference in the plasma drug levels of clorazepate mono- and dipotassium salts and the metabolite was found in dogs.

  3. Clinical significance of plasma level of AT-Ⅲ determination in sepsis patients

    Institute of Scientific and Technical Information of China (English)

    Wei Chen; Zhi-Hua Hu; Chen-Mian

    2015-01-01

    Objective:Through measure changes of anticoagulant enzyme (AT-Ⅲ) activity in plasma in sepsis patients, this paper discusses the clinical significance of AT-Ⅲ activity changes in predicting sepsis occurrence and prognosis.Methods: The non-sepsis 30 cases, with sepsis 76 cases, including 25 cases of severe sepsis, use method of thrombin gelatum lacuna for determining activity of AT-Ⅲ in plasma, platelet count and APACHEⅢ score simultaneously. Results:Sepsis group, severe sepsis groups contrast with the non-sepsis group respectively, activity of AT-Ⅲ reduced significantly (P<0.01), severe sepsis group lower than sepsis group (P=0.055).Conclusion:AT-Ⅲ activity reduced early in sepsis patients, with patient's condition aggravat, its value further reduced, hints measurement of AT-Ⅲ activity has certainly clinical significance in predicting sepsis occurrence and prognosis.

  4. Quantitative determination of amisulpride in rat plasma by HPLC-MS/MS.

    Science.gov (United States)

    Noh, Keumhan; Jang, Yoo-Jeong; Kwon, Kwang-il; Kim, Eunyoung; Jeong, Tae Cheon; Yun, Hwi-yeol; Kang, Wonku

    2015-01-01

    Amisulpride, a selective antagonist of D2 and D3 dopamine receptors, is used as an antipsychotic drug. In this study, we reported a sensitive LC-MS/MS method for determining amisulpride concentrations in rat plasma, and a preclinical pharmacokinetic study in the rat. After a simple protein precipitation with acetonitrile containing methaqualone as an internal standard, the analytes were separated on a reversed-phase column with a mobile phase of 0.2 % aqueous formic acid and acetonitrile (3:7, v/v). The accuracy and precision of the assay were in accordance with FDA guidance for the validation of bioanalytical methods. This analytical method was used successfully to characterize the time course of the plasma concentration of amisulpride following oral administration of a single 10 mg/kg dose in rats.

  5. RP-HPLC Method Development and Its Validation for Quantitative Determination of Rimonabant in Human Plasma

    Directory of Open Access Journals (Sweden)

    Shravan Bankey

    2012-01-01

    Full Text Available A simple, accurate, and precise HPLC method was developed and validated for determination of rimonabant in human plasma. Following liquid-liquid extraction, chromatographic separation was accomplished using C18 column with mobile phase consisting of acetonitrile : water (90 : 10, v/v, drug was detected at 260 nm using UVdetector. The LOD and LOQ were 3.0 and 10.0 μg/L, respectively. The method is linear in the interval 50.0–1000.0 μg/L. The average extraction recovery of drug from plasma was found to be 92.2%. The percent CV of the method was found to be less than 10.8%, and accuracy was found between 94.5 and 106.7%. The assay may be applied to a pharmacokinetic and bioequivalence study of rimonabant.

  6. Quantitative determination of citric acid in seminal plasma by using Raman spectroscopy.

    Science.gov (United States)

    Huang, Zufang; Chen, Xiwen; Li, Yongzeng; Chen, Jinhua; Lin, Juqiang; Wang, Jing; Lei, Jinping; Chen, Rong

    2013-07-01

    In this study, Raman spectroscopy was first used to study the linear relationship between Raman spectral intensities and citric acid concentrations in aqueous solution. By using the specific Raman band of 942 cm(-1), concentrations of citric acid ranging from 2 to 20 mg/mL were observed linearly (R(2) = 0.993), and the limit of detection was 1.0 mg/mL. Then, citric acid detection in clinical seminal plasma ultrafiltrate samples was performed, and the intensity of the Raman-specific peak demonstrates a good linear correlation (R(2) = 0.946) with citric acid concentrations determined by the enzymatic method. Our results showed that Raman spectroscopy has the potential of being applied to detect concentrations of citric acid in seminal plasma in clinic.

  7. Determination of structure tilting in magnetized plasmas - Time delay estimation in two dimensions

    CERN Document Server

    Guszejnov, Dávid; Zoletnik, Sándor; Andreas-Krämer-Flecken,

    2013-01-01

    Time delay estimation (TDE) is a well-known technique to investigate poloidal flows in fusion plasmas. The present work is an extension of the earlier works of A. Bencze and S. Zoletnik 2005 and B. T\\'al et al. 2011. From the prospective of the comparison of theory and experiment it seem to be important to estimate the statistical properties of the TDE based on solid mathematical groundings. This paper provides analytic derivation of the variance of the TDE using a two-dimensional model for coherent turbulent structures in the plasma edge and also gives an explicit method for determination of the tilt angle of structures. As a demonstration this method is then applied to the results of a quasi-2D Beam Emission Spectroscopy (BES) measurement performed at the TEXTOR tokamak.

  8. [Rapid determination of propranolol enantiomers in rat plasma by column-switching-high performance liquid chromatography].

    Science.gov (United States)

    Wu, Xiaoyu; Wang, Rong; Xie, Hua; Wang, Jianfeng; Jia, Zhengping; Zhang, Qiang; Wang, Xianhua

    2011-12-01

    A high performance liquid chromatographic (HPLC) method with column-switching was developed and validated for rapid determination of two propranolol enantiomers in rat plasma. The column of restricted-access media was used as a pre-treatment column and a Chiralcel OD-RH was used as analytical column. The plasma samples were injected directly into the pretreatment column to remove plasma protein and endogenous constituents as well as to retent the propranolol enantiomers in the column using the mobile phase of borate buffer (pH 8.5)-methanol (95:5, v/v) at the flow rate of 1.0 mL/min. Then the propranolol enantiomers were transferred to the Chiralcel OD-RH column using the mobile phase of isopropanol-ethanol-0.2 mmol/L borate buffer (pH 8.5) (30: 30: 40, v/v/v) at a flow rate of 0.8 mL/min by column-switching technology. The column-switching time was 3.0 min, the used wavelength was 293 nm and the column temperature was set at 25 degrees C. The calibration curve showed excellent linear relationship (r = 0.999 5) in the concentration range from 25 mg/L to 500 mg/L for propranolol enantiomers in plasma. The intra-day and inter-day assay precisions and accuracies were well and the relative standard deviations (RSDs) were less than 5%. The average recoveries (n = 6) of the two enantiomers at 3 spiked levels were from 97.89% to 101.56%. All the values of the method validation were within the generally accepted criteria for biological sample analysis. The results show that the method is convenient, quick, sensitive and accurate. The method was successfully applied in the determination of propranolol enantiomers in rat blood pharmacokinetics study.

  9. Control plasma renin activity and changes in sympathetic tone as determinants of minoxidil-induced increase in plasma renin activity.

    Science.gov (United States)

    O'Malley, K; Velasco, M; Wells, J; McNay, J L

    1975-01-01

    A study was made of the possible mechanism(s) underlying minoxidil-induced increase in plasma renin activity (PRA). 10 patients with essential hypertension were treated with minoxidil and subsequently with a combination of minoxidil plus propranolol. Minoxidil lowered mean arterial pressure 31.6 plus or minus 3.3 mm Hg, mean plus or minus SEM. There was an associated increase in both PRA, 6.26 plus or minus 2.43 NG/ML/H, and heart rate, 21.4 plus or minus 2.7 beats/min. The changes in PRA and heart rate were positively correlated, r, 0.79. Addition of propranolol reduced mean arterial pressure by a further 10.1 plus or minus 1.5 mm Hg and returned heart rate to control levels. Propranolol reduced PRA significantly but not to control levels. Control PRA positively correlated with PRA on minoxidil, r, 0.97, and with PRA on minoxidil plus propranolol, r, 0.98. We conclude that control PRA is a major determinant of change in PRA with minoxidil. Minoxidil increased PRA by at least two mechanisms: (a) an adrenergic mechanism closely related to change in heart rate and blocked by propranolol, and (b) a mechanism(s) not sensitive to propranolol and possibly related to decrease in renal perfusion pressure. PMID:1127099

  10. Application of poloidal beta and plasma internal inductance in determination of input power time of Damavand tokamak

    Science.gov (United States)

    Noori, Ehsanallah; Sadeghi, Yahya; Ghoranneviss, Mahmood

    2016-10-01

    In this study, magnetic measurement of poloidal fields were used to determine poloidal beta and plasma internal inductance of Damavand tokamak combination of poloidal beta and plasma internal inductance (β _p+{l_i}/{2} ), known as Shafranov parameter, was obtained experimentally in terms of normal and tangential components of the magnetic field. Plasma internal inductance and poloidal beta were obtained using parametrization method based on analytical solution of Grad-Shafranov equation (GSE) and compared with parabolic-like profile of toroidal current density approach for determination of the plasma internal inductance. Finding evolution of β _p+{l_i}/{2} and plasma internal inductance. Finding poloidal beta (Shafranov parameter and internal inductance) and using energy balance equation, thermal energy and energy confinement were determined qualitatively in terms of poloidal beta during a regular discharge of Damavand tokamak.

  11. Dependence of LTX plasma performance on surface conditions as determined by in situ analysis of plasma facing components

    Energy Technology Data Exchange (ETDEWEB)

    Lucia, M., E-mail: mlucia@pppl.gov [Princeton Plasma Physics Laboratory (PPPL), Princeton, NJ 08543 (United States); Kaita, R.; Majeski, R. [Princeton Plasma Physics Laboratory (PPPL), Princeton, NJ 08543 (United States); Bedoya, F.; Allain, J.P. [University of Illinois at Urbana-Champaign (UIUC), Urbana, IL 61801 (United States); Abrams, T.; Bell, R.E.; Boyle, D.P.; Jaworski, M.A.; Schmitt, J.C. [Princeton Plasma Physics Laboratory (PPPL), Princeton, NJ 08543 (United States)

    2015-08-15

    The Materials Analysis and Particle Probe (MAPP) diagnostic has been implemented on the Lithium Tokamak Experiment (LTX) at PPPL, providing the first in situ X-ray photoelectron spectroscopy (XPS) surface characterization of tokamak plasma facing components (PFCs). MAPP samples were exposed to argon glow discharge conditioning (GDC), lithium evaporations, and hydrogen tokamak discharges inside LTX. Samples were analyzed with XPS, and alterations to surface conditions were correlated against observed LTX plasma performance changes. Argon GDC caused the accumulation of nm-scale metal oxide layers on the PFC surface, which appeared to bury surface carbon and oxygen contamination and thus improve plasma performance. Lithium evaporation led to the rapid formation of a lithium oxide (Li{sub 2}O) surface; plasma performance was strongly improved for sufficiently thick evaporative coatings. Results indicate that a 5 h argon GDC or a 50 nm evaporative lithium coating will both significantly improve LTX plasma performance.

  12. Betaine as a determinant of postmethionine load total plasma homocysteine before and after B-vitamin supplementation.

    NARCIS (Netherlands)

    Holm, P.I.; Bleie, O.; Ueland, P.M.; Lien, E.A.; Refsum, H.; Nordrehaug, J.E.; Nygard, O.

    2004-01-01

    OBJECTIVE: Betaine is a substrate in the betaine-homocysteine methyltransferase reaction, converting homocysteine to methionine. There are only sparse data on plasma betaine as a determinant of the plasma total homocysteine (tHcy) concentration. METHODS AND RESULTS: Ninety patients undergoing corona

  13. QUANTITATIVE-DETERMINATION OF THE DOPAMINE AGONIST LISURIDE IN PLASMA USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUORESCENCE DETECTION

    NARCIS (Netherlands)

    WOLTHERS, BG; KAMERBEEK, WDJV; VANBEUSEKOM, CM; ELSHOF, F; BUITENHUIS, AWD; BRUNT, EPR; LAKKE, JPWF

    1993-01-01

    An HPLC method for the determination of lisuride hydrogen maleate in plasma is described. After addition of ergotamine tartrate as internal standard, plasma is extracted with diethyl ether. Following evaporation of the solvent and redissolving in methanol the extract is injected on a silica HPLC col

  14. The influence of progesterone alone and in combination with estradiol on ventricular action potential duration and triangulation in response to potassium channel inhibition.

    Science.gov (United States)

    Tisdale, James E; Overholser, Brian R; Wroblewski, Heather A; Sowinski, Kevin M

    2011-03-01

    Females are at increased risk for torsades de pointes (TdP). Some evidence suggests that progesterone may protect against TdP, but few data exist regarding the effects of progesterone on cardiac repolarization. We determined the effects of progesterone alone and in combination with estradiol on ventricular action potential duration (APD) and triangulation in response to potassium channel inhibition. Female New Zealand white rabbits (n = 30) underwent ovariectomy and were implanted with 21-day sustained release pellets (each n = 6): progesterone; estradiol; progesterone; & estradiol combined; dihydrotestosterone (DHT); and placebo. After 20 days, hearts were excised, mounted, perfused with modified Krebs-Henseleit buffer, and paced at 150 bpm. After baseline measurements, hearts were perfused with quinidine 3 μmol/L. The degree of quinidine-associated prolongation of ventricular APD at 90% repolarization (APD(90) ) in the progesterone group was significantly less than that in the estradiol and the combined estradiol and progesterone groups, and not significantly different than in the DHT group. The degree of prolongation of action potential triangulation (APD(90) - APD(30) ) in hearts from progesterone-treated rabbits was significantly less than that in the estradiol group, and not significantly different from that in hearts from DHT-treated rabbits. There were no significant differences in quinidine effects on ventricular APD(90) or action potential triangulation between hearts exposed to estradiol alone or those exposed to both estradiol and progesterone. Progesterone protects against prolongation of APD(90) and triangulation associated with potassium channel inhibition. However, progesterone does not attenuate the effects of estradiol on prolongation of ventricular APD(90) associated with potassium channel inhibition. © 2010 Wiley Periodicals, Inc.

  15. Determination of plasma protein synthesis index in liver diseases by means of /sup 75/Se-selenomethionine

    Energy Technology Data Exchange (ETDEWEB)

    Pirwitz, B. (Centrum Medyczne Ksztalcenia Podyplomowego, Warsaw (Poland))

    1979-01-01

    The investigations were carried out in 96 patients with chronic liver diseases and 44 controls. After intravenous administration of /sup 75/Se-selenomethionine plasma and plasma-protein radioactivity was measured at intervals of 2 hours. On the basis of these measurements the index of plasma protein synthesis rate was determined. It was found that the index of plasma protein synthesis in liver cirrhosis was significantly decreased in an overwhelming number of cases in relation to controls. On the other hand, in liver neoplasms this index was statistically significantly increased. It is possible that this fact will be used in future for differential diagnosis.

  16. Effects of progesterone withdrawal on uterine secretion of prostaglandin F2 alpha in response to oxytocin in ewes.

    Science.gov (United States)

    Kaminski, M A; Hayes, S H; Silvia, W J

    1997-01-01

    Two experiments were conducted to determine if withdrawal of progesterone during the luteal phase of the oestrous cycle affected the ability of the ovine uterus to secrete prostaglandin F2 alpha (PGF2 alpha) in response to oxytocin. In Experiment 1, 18 ewes were ovariectomized on Day 9 and Day 12 after oestrus. Ewes were subdivided into three treatment groups (n = 6 per group): Group-1 ewes underwent sham surgery; Group-2 ewes received oestradiol (OVX + O); and Group-3 ewes received oestradiol + progesterone (OVX + O,P). Oxytocin was administered to each ewe on Days 10, 13 and 15 after oestrus. Concentrations of 13, 14-dihydro-15-keto-PGF2 alpha (PGFM) were determined in samples of jugular venous blood for 2 h after oxytocin challenge. The magnitude of the PGFM response 24 h after ovariectomy was greater (P Day 11 and assigned to 1 of 4 treatment groups (n = 6 per group): Group 1, no steroid replacement (OVX); Group 2, oestradiol replacement (OVX + O); Group 3, progesterone replacement (OVX + P); or Group 4, progesterone + oestradiol replacement (OVX + O,P). Ewes received oxytocin on Day 12 and Day 15. On Day 12, uterine secretory responsiveness to oxytocin was greatest in ewes in the OVX + O group (P < 0.1). Responsiveness was low in ewes in the OVX group, as it was in ewes in both groups that received progesterone replacement. Therefore, the increase in uterine secretory responsiveness to oxytocin following progesterone withdrawal is dependent on oestradiol replacement.

  17. Determination of Trace Elements in High Purity Gold by High Resolution Inductively Coupled Plasma Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    XIE Hualin; HUANG Kelong; NIE Xidu; FU Liang

    2009-01-01

    Trace elements were determined in high purity gold by high resolution inductively coupled plasma mass spectrometry(HR-ICP-MS).Sample were decomposed by aqua regia.To overcome some potentially problematic spectral interference,measurements were acquired in both medium and high resolution modes.The matrix effects due to the presence of excessive HCl and Au were evaluated.The optimum conditions for the determination was tested and discussed.The standard addition method was employed for quantitative analysis.The detection limits range from 0.01 μg/g to 0.28 μg/g depending on the elements.The method is accurate,quick and convenient.It has been applied to the determination of trace elements in high purity gold with satisfactory results.

  18. Inductively coupled plasma optical emission spectroscopy determination of trace element composition of argan oil.

    Science.gov (United States)

    Gonzálvez, A; Ghanjaoui, M E; El Rhazi, M; de la Guardia, M

    2010-02-01

    A methodology based on inductively coupled plasma optical emission spectroscopy (ICP-OES) after microwave assisted acid digestion has been developed to determine the trace element content of Moroccan argan oil. Limit of detection values equal or lower than few mg/kg were obtained for all elements under study. To assure the accuracy of the whole procedure, recovery studies were carried out on argan oil samples spiked at different concentration levels from 10 to 200 µg/L. Quantitative average recovery values were obtained for all elements evaluated, demonstrating the suitability of this methodology for the determination of trace elements in argan oil samples. Aluminum, calcium, chromium, iron, potassium, lithium, magnesium, sodium, vanadium and zinc were quantitatively determined in Moroccan argan oils being found that their concentration is different of that found in other edible oils thus offering a way for authentication and for the evaluation of possible adulterations.

  19. Progesterone receptor membrane component 1 deficiency attenuates growth while promoting chemosensitivity of human endometrial xenograft tumors

    NARCIS (Netherlands)

    A.M. Friel (Anne M.); L. Zhang (Ling); C.A. Pru (Cindy A.); N.C. Clark (Nicole C.); M.L. McCallum (Melissa L.); L.J. Blok (Leen); T. Shioda (Toshi); J.J. Peluso (John J.); B.R. Rueda (Bo R.); J.K. Pru (James K.)

    2015-01-01

    textabstractEndometrial cancer is the leading gynecologic cancer in women in the United States with 52,630 women predicted to be diagnosed with the disease in 2014. The objective of this study was to determine if progesterone (P4) receptor membrane component 1 (PGRMC1) influenced endometrial cancer

  20. Roles of estrogen and progesterone in modulating renal nerve function in the rat kidney

    Directory of Open Access Journals (Sweden)

    J.B. Graceli

    2013-07-01

    Full Text Available The maintenance of extracellular Na+ and Cl- concentrations in mammals depends, at least in part, on renal function. It has been shown that neural and endocrine mechanisms regulate extracellular fluid volume and transport of electrolytes along nephrons. Studies of sex hormones and renal nerves suggested that sex hormones modulate renal function, although this relationship is not well understood in the kidney. To better understand the role of these hormones on the effects that renal nerves have on Na+ and Cl- reabsorption, we studied the effects of renal denervation and oophorectomy in female rats. Oophorectomized (OVX rats received 17β-estradiol benzoate (OVE, 2.0 mg·kg-1·day-1, sc and progesterone (OVP, 1.7 mg·kg-1·day-1, sc. We assessed Na+ and Cl- fractional excretion (FENa+ and FECl- , respectively and renal and plasma catecholamine release concentrations. FENa+ , FECl- , water intake, urinary flow, and renal and plasma catecholamine release levels increased in OVX vs control rats. These effects were reversed by 17β-estradiol benzoate but not by progesterone. Renal denervation did not alter FENa+ , FECl- , water intake, or urinary flow values vs controls. However, the renal catecholamine release level was decreased in the OVP (236.6±36.1 ng/g and denervated rat groups (D: 102.1±15.7; ODE: 108.7±23.2; ODP: 101.1±22.1 ng/g. Furthermore, combining OVX + D (OD: 111.9±25.4 decreased renal catecholamine release levels compared to either treatment alone. OVE normalized and OVP reduced renal catecholamine release levels, and the effects on plasma catecholamine release levels were reversed by ODE and ODP replacement in OD. These data suggest that progesterone may influence catecholamine release levels by renal innervation and that there are complex interactions among renal nerves, estrogen, and progesterone in the modulation of renal function.

  1. [Simultaneous determination of adrenaline, noradrenaline, cortisone and cortisol in plasma with HPLC/MS/MS].

    Science.gov (United States)

    Yong, Li; Wang, Yu; Zou, Xiao-Li; Zhu, Lan; Xie, Hui-Ru; Li, Long-Jiang

    2014-01-01

    To develop a method for simultaneous determination of adrenaline, noradrenaline, cortisone and cortisol in plasma using HPLC/MS/MS. Sample proteins were precipitated with acetonitrile and the sample solution was injected into HPLC/MS/MS after centrifugation at 15,000 r/min for 5 min. Electrospray ionization (ESI) and the positive ion detection were applied with a multiple reaction monitoring (MRM) mode for quantitative analyses. Under the optimal conditions, good linearity (r > 0.999) was observed in the range of 0.02-200.00 ng/mL of target compounds. The detection limit reached 4.13 pg/mL, 4.64 pg/mL, 4.29 pg/mL and 4.52 pg/mL for adrenaline, noradrenaline, cortisone and cortisol respectively. The inter-day and intra-day precisions ranged from 1.19%-5.42% and 2.16%-6.04% respectively. Satisfied results were achieved using human plasma samples, with a spiked recovery in the range of 80.0%-109.0% and a relative standard deviation of 3.93%-7.57%. The proposed method is quick, sensitive and suitable for batch analyses of plasma samples.

  2. Liquid Chromatography-Electrospray Ionization Mass Spectrometry Method for Determination of Protopanaxadiol in Rat Plasma

    Institute of Scientific and Technical Information of China (English)

    WU Yi; TENG Guo-sheng; LIANG Yong-tao; ZHONG Da-fang; LIU Bing

    2009-01-01

    A simple, rapid and sensitive method for the determination of protopanaxadiol in rat plasma with ginse-noside Rh2 as internal standard was developed and validated. The analyte and internal standard were extracted from plasma with ether-dichloromethane(3:2, volume ratio) and then were analyzed by reversed-phase HPLC on a short Zorbax Extend C_(18) column(50 mm×2.1 mm, 3.5 μm i.d.) eluted with a mobile phase consisting of acetoni-trile/methanol 0.10 mmol/L ammonium acetate(45:45:10, volume ratio) at 0.4 mL/min. Detection was performed on an Applied Biosystems Sciex API 4000 mass spectrometer set at unit resolution in the multiple reaction monitoring mode. Electrospray ionization was used for ion production. The assay method shows linear over a range of 5-2000 ng/mL and intra- and inter-day precisions over this range were <10.0% with accuracy ranged from 86.3% to 114.1%. The limit of detection was 500 pg/mL in the plasma. The method was successfully applied to a preclinical pharmaco-kinetic study of protopanaxadiol(17.5 mg/kg) administered as a single oral dose.

  3. Determination of moxonidine in human plasma by liquid chromatography-electrospray ionisation-mass spectrometry.

    Science.gov (United States)

    Zhao, Luhua; Ding, Li; Wei, Xin

    2006-01-23

    A sensitive and specific liquid chromatography-electrospray ionisation-mass spectrometry (LC-ESI-MS) method has been developed and validated for the identification and quantification of moxonidine in human plasma. After the addition of clonidine-HCl, the internal standard (IS) and sodium hydrogen carbonate, plasma samples were extracted using 5 mL ethyl acetate. The compounds were separated on a Lichrospher ODS (5 microm, 250 mm x 4.6 mm) column using an elution system of 10 mmol/L ammonium acetate buffer-methanol (20:80 v/v) as the mobile phase. Analytes were determined using electrospary ionization in a single quadrupole mass spectrometer. LC-ESI-MS was performed in the selected-ion monitoring (SIM) mode using target ions at m/z: 242.2 for moxonidine and m/z: 230.1 for the IS. The method has shown to be sensitive and specific by testing six different blank plasma batches. Linearity was established for the range of concentrations 0.01976-9.88 ng/mL with a coefficient of correlation (r) of 0.9999. The lower limit of quantification (LOQ) was identifiable and reproducible at 0.01976 ng/mL. The method has been successfully applied to study the pharmacokinetics of moxonidine in healthy male Chinese volunteers.

  4. Determination of Duloxetine and Its Major Metabolites in Rabbit Plasma by High-Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    T. K. Laha

    2015-03-01

    Full Text Available A rapid and sensitive high performance liquid chromatographic method is described for simultaneous determination of duloxetine and its major metabolites, such as 4- hydroxy duloxetine (M7, Glucuronide conjugate of 5-hydroxy-6-methoxy duloxetine (M6 and Glucuronide conjugate of dihydrodiol duloxetine (M12 in rabbit plasma. HPLC analysis was carried out on a µ-Bondapak C18 column (250 mm × 4.6 mm, 5µm particle size using methanol: phosphate buffer (pH 7.9, 50 mM (7:3 v/v as the mobile phase at a flow rate of 1.5 ml/min. Detection was carried out at 224 nm with an UV detector. The above metabolites present in the rabbit plasma were characterized by retro-synthesis followed by NMR and MS study for structure confirmation and finally injected separately into the HPLC system. All the three retention time matches with the metabolites present in the plasma sample.

  5. Sensitive determination of ranitidine in rabbit plasma by HPLC with fluorescence detection.

    Science.gov (United States)

    Khedr, Alaa

    2008-02-01

    A sensitive high-performance liquid chromatographic method for determination of ranitidine (RAN) in rabbit plasma is described. The method is based on liquid-liquid extraction, labeling with dansyl chloride and monitoring with fluorescence detector at 338nm (ex)/523nm (em). Plasma samples were extracted with diethyl ether alkalinized with 1M sodium hydroxide. Ephedrine HCl (EPH-HCl) was used as internal standard. Both, RAN and EPH were completely derivatized after heating at 60 degrees C for 10min in sodium bicarbonate solution (pH 9.5). The derivatized samples were analyzed by HPLC using Agilent Zorbax Extended C18 column (150mmx4.6mm i.d.) and mobile phase consists of 48% acetonitrile and 52% sodium acetate solution (0.02M, pH 4.6). The linearity of the method was in the range of 0.025-10microg/ml. The limits of detection (LOD) and quantification (LOQ) were 7.5+/-0.18 and 22.5+/-0.12ng/ml, respectively. Ranitidine recovery was 97.5+/-1.1% (n=6; R.S.D.=1.8%). The method was applied on plasma collected from rabbits at different time intervals after oral administration of 5mg/kg ranitidine HCl.

  6. Determination and pharmacokinetic study of catechin in rat plasma by HPLC

    Institute of Scientific and Technical Information of China (English)

    Li Xie; Xin-Nan Li; De-Xi Jiang; Dan Zhang

    2011-01-01

    A high performance liquid chromatographic method was developed and validated for the quantitative determination of catechin in rat plasma and its pharmacokinetic study after intragastric administration of Catechu and Xiongdanjiangre Wan into SD rats. Plasma samples were prepared by protein precipitation using methanol-5% aqueous zinc sulfate (70:30, v/v) as precipitant. Chromatographic separation was achieved on Hypersil Cl8 column (250 mm~ 4.6 mm, 10 pm) with acetonitrile-water-triethylamine (6:94:0.3, v/v/v, pH 4.0+0.1, adjusted with phos- phoric acid) as mobile phase, followed by a UV detection at 207 nm. Good linearity was obtained over the range of 0.143-7.15 mg/L of catechin, with correlation coefficient of 0.9992. The method was simple, sensitive, accurate and reproducible and' has been successfully applied to the pharmacokinetic study of catechin in rat plasma.

  7. Determination of quinolones in plasma samples by capillary electrophoresis using solid-phase extraction.

    Science.gov (United States)

    Hernández, M; Borrull, F; Calull, M

    2000-06-09

    The potential of capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MEKC) have been investigated for the separation and quantitative determination of 10 quinolone antibiotics. The influence of different conditions, such as the buffer and pH of the electrolyte, the surfactant and the ion-pairing agents added to the electrolyte and the organic modifier were studied. A buffer consisting of 40 mM sodium tetraborate at pH 8.1 containing 10% (v/v) methanol was found to be a highly efficient electrophoretic system for separating lomefloxacin, enoxacin, norfloxacin, pipemidic acid, ofloxacin, piromidic acid, flumequine, oxolinic acid, cinoxacin and nalidixic acid. A solid-phase extraction method to remove the sample matrix (pig plasma samples) was developed on a C(18) cartridge using a mixture of methanol-water (70:30, v/v). The method is specific and reproducible and mean recoveries were in the range 94.0+/-4.2% and 123.3+/-4.1% for pig plasma samples over the range used. A linear relationship between concentration and peak area for each compound in pig plasma samples was obtained in the concentration range 5-20 mg l(-1) and detection limits were between 1.1 and 2.4 mg l(-1).

  8. Determination of tolperisone in human plasma by liquid chromatography/tandem mass spectrometry for clinical application.

    Science.gov (United States)

    Choi, Chang-Ik; Park, Jung-In; Lee, Hye-In; Lee, Yun-Jeong; Jang, Choon-Gon; Bae, Jung-Woo; Lee, Seok-Yong

    2012-12-12

    We have developed and validated a simple, rapid, and sensitive liquid chromatography analytical method employing tandem mass spectrometry (LC-MS/MS) for the determination of tolperisone, a centrally acting muscle relaxant, in human plasma. After liquid-liquid extraction with methyl t-butyl ether, chromatographic separation of tolperisone was performed using a reversed-phase Luna C(18) column (2.0mm×50mm, 5μm particles) with a mobile phase of 10mM ammonium formate buffer (pH 3.5) - methanol (12:88, v/v) and quantified by tandem mass detection in ESI positive ion mode. The flow rate of the mobile phase was 250μL/min and the retention times of tolperisone and the internal standard (IS, dibucaine) were both 0.6min. The calibration curves were linear over a range of 0.5-300ng/mL (r>0.999). The lower limit of quantification, using 200μL human plasma, was 0.5ng/mL. The mean accuracy and precision for intra- and inter-day validation of tolperisone were within acceptable limits. The LC-MS/MS method reported here showed improved sensitivity for quantification of tolperisone in human plasma compared with previously described analytical methods. Lastly, the validated method was successfully applied to a pharmacokinetic study in humans.

  9. Determination of trovafloxacin and marbofloxacin in sheep plasma samples by HPLC using UV detection.

    Science.gov (United States)

    Mahmood, Adnan H; Medley, Gregory A; Grice, Jeffrey E; Liu, Xin; Roberts, Michael S

    2012-03-25

    A simple and rapid high performance liquid chromatographic method was developed, validated and applied for the simultaneous determination of marbofloxacin (MBX) and trovafloxacin (TVX) in sheep plasma. Samples were extracted with 20% perchloric acid and MBX and TVX were separated on a C(18) column using a gradient mobile phase system consisting of 17.5mM of NaH(2)PO(4) and 1.5mM of tetrabutylammonium hydroxide aqueous solution, pH 3 (A) and 50% acetonitrile and 50% methanol (B), with UV detection at 293 and 270 nm. The retention times of MBX and TVX were 4.9 and 6.6 min respectively. The detection and quantification limits for MBX and TVX were 2 ng/mL and 10 ng/mL respectively for both compounds. The calibration curves were linear over a concentration range of 10-50,000 ng/mL for both antibiotics. The linearity, precision, accuracy, recovery and stability of the assay were evaluated from spiked sheep plasma. The method was successfully applied to sheep plasma samples obtained from MBX and TVX pharmacokinetic studies.

  10. DETERMINATION OF PROMINENCE PLASMA {beta} FROM THE DYNAMICS OF RISING PLUMES

    Energy Technology Data Exchange (ETDEWEB)

    Hillier, Andrew [Kwasan and Hida Observatories, Kyoto University, Kyoto (Japan); Hillier, Richard [Department of Aeronautics, Imperial College, London (United Kingdom); Tripathi, Durgesh, E-mail: andrew@kwasan.kyoto-u.ac.jp [Inter-University Centre for Astronomy and Astrophysics, Post Bag 4, Ganeshkhind, Pune 411 007 (India)

    2012-12-20

    Observations by the Hinode satellite show in great detail the dynamics of rising plumes, dark in chromospheric lines, in quiescent prominences that propagate from large ({approx}10 Mm) bubbles that form at the base of the prominences. These plumes present a very interesting opportunity to study magnetohydrodynamic (MHD) phenomena in quiescent prominences, but obstacles still remain. One of the biggest issues is that of the magnetic field strength, which is not easily measurable in prominences. In this paper we present a method that may be used to determine a prominence's plasma {beta} when rising plumes are observed. Using the classic fluid dynamic solution for flow around a circular cylinder with an MHD correction, the compression of the prominence material can be estimated. This has been successfully confirmed through simulations; application to a prominence gave an estimate of the plasma {beta} as {beta} = 0.47 {+-} 0.079 to 1.13 {+-} 0.080 for the range {gamma} = 1.4-1.7. Using this method it may be possible to estimate the plasma {beta} of observed prominences, therefore helping our understanding of a prominence's dynamics in terms of MHD phenomena.

  11. [Isolation, extractive concentration, and determination of caffeine in the studies of blood plasma].

    Science.gov (United States)

    Korenman, Ia I; Shormanov, V K; Mokshina, N Ia; Krivosheeva, O A; Golubitskiĭ, G B

    2012-01-01

    The optimal conditions for the isolation of caffeine from human blood by means of acetone extraction are described with special reference to the peculiarities of extraction from aqueous solutions. The possibility of concentration and purification of caffeine from blood plasma using acetone and aceton-chlorophorm mixture (2:8) as the solvents is illustrated. In addition, purification by silica-gel thin layer chromatography is discussed. Thin layer chromatography, UV-spectrophotometry, and high performance liquid chromatography are considered as potential methods for the identification and quantitative determination of caffeine.

  12. Determination of rhoifolin and daidzin in human plasma by high-performance liquid chromatography.

    Science.gov (United States)

    Ishii, K; Urano, S; Furuta, T; Kasuya, Y

    1994-05-13

    A method for determining flavonoids in human plasma is presented for application to pharmacokinetic studies of two flavonoids, rhoifolin and daidzin. Isocratic reversed-phase high-performance liquid chromatography (HPLC) was used with genistin as an internal standard and solid-phase extraction using a Sep-Pak C18 cartridge. The mobile phases were acetonitrile-0.1 M ammonium acetate solution (20:80, v/v) for rhoifolin and methanol-0.1 M ammonium acetate solution (33:67, v/v) for daidzin. The detection limits on-column were 2 ng for rhoifolin and 0.5 ng for daidzin.

  13. Inductively Coupled Plasma Optical-Emission Spectroscopy Determination of Major and Minor Elements in Vinegar

    Directory of Open Access Journals (Sweden)

    Arzu AKPINAR-BAYIZIT

    2010-12-01

    Full Text Available This study characterizes the mineral content of vinegar samples. The concentrations of Na, K, Ca, Mg and P (major elements as well as Fe, Mn, Sn, Cu, Ni, Zn, Pb and Cd (minor elements were determined in 35 commercial vinegar samples using inductively coupled plasma optical-emission spectrometry (ICP-OES. The elements with the highest concentrations were K, Na, Ca, Mg and P. The concentrations of heavy metals in the vinegar samples, including Cd, Ni, Sn and Pb, were not considered a health risk.

  14. [Determination of total sulfur in coal by inductively coupled plasma atomic emission spectrometry].

    Science.gov (United States)

    Liu, Dong-yan; Zhang, Yuan-li

    2002-02-01

    A direct method was reported for the determination of total sulfur in coal by inductively coupled plasma atomic emission spectrometry (ICP-AES). The dissolution conditions of coal samples as well as interference conditions of hydrochloric acid and matrix were studied. The recommended method not only proved to be simple and rapid than traditional gravimetric method but show satisfying precision and accuracy as well. The results of samples are as same as gravimetry. The recoveries are more than 96%, and the relative standard deviation of six samples are less than 3%.

  15. Iodine Determination by Microwave Plasma Torch Atomic Emission Spectrometer Coupled with Online Preconcentration Vapor Generation Technique

    Institute of Scientific and Technical Information of China (English)

    FEI Yan-qun; LUO Gui-min; FENG Guo-dong; CHEN Huan-wen; FEI Qiang; HUAN Yan-fu; JIN Qin-han

    2008-01-01

    This article focuses on iodine determination by microwave plasma torch atomic emission spectrometry (MPT-AES) coupled with online preconcentration vapor generation method.A new desolvation device,multistrand Nation dryer,was used as the substitute for condenser desolvation system.Some experimental conditions,such as preconcentration time,acidity of sample solution,rinsing solution acidity and dynamic linear range were investigated and optimized.The new desolvation system eliminates the problem of decreasing emission intensity of I(I) 206.238 nm line with the increase of working time on a conventional condenser desolvation system,thus greatly improving the reproducibility.

  16. Determination of Free Levels of Cinitipride in Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Shikha M. N. Roy

    2008-01-01

    4.6 mm, 5 µm i.d.. The assay of cinitapride was linear calibration curve over the range 20.118 pg mL−1 to 2011.797 pg mL−1. Plasma concentrations of cinitapride were determined by LC-MS/MS with a limit of quantification of 20.118 pg mL−1 that allowed an appropriate characterization of the pharmacokinetic profile of cinitapride at the therapeutic dose. The method was successfully applied to the bioequivalence study of cinitapride tablet (1.0 mg administered as a single oral dose.

  17. Analyses of the plasma generated by laser irradiation on sputtered target for determination of the thickness used for plasma generation

    Energy Technology Data Exchange (ETDEWEB)

    Kumaki, Masafumi, E-mail: masafumi.kumaki@riken.jp [Cooperative Major in Nuclear Energy, Waseda University, Shinjuku, Tokyo (Japan); RIKEN, Wako, Saitama (Japan); Ikeda, Shunsuke; Sekine, Megumi; Munemoto, Naoya [RIKEN, Wako, Saitama (Japan); Department of Energy Sciences, Tokyo Institute of Technology, Meguro, Tokyo (Japan); Fuwa, Yasuhiro [RIKEN, Wako, Saitama (Japan); Department of Physics and Astronomy, Kyoto University, Uji, Kyoto (Japan); Cinquegrani, David [American Nuclear Society, University of Michigan, Ann Arbor, Michigan 48109 (United States); Kanesue, Takeshi; Okamura, Masahiro [Collider-Accelerator Department, Brookhaven National Laboratory, Upton, New York 11973 (United States); Washio, Masakazu [Cooperative Major in Nuclear Energy, Waseda University, Shinjuku, Tokyo (Japan)

    2014-02-15

    In Brookhaven National Laboratory, laser ion source has been developed to provide heavy ion beams by using plasma generation with 1064 nm Nd:YAG laser irradiation onto solid targets. The laser energy is transferred to the target material and creates a crater on the surface. However, only the partial material can be turned into plasma state and the other portion is considered to be just vaporized. Since heat propagation in the target material requires more than typical laser irradiation period, which is typically several ns, only the certain depth of the layers may contribute to form the plasma. As a result, the depth is more than 500 nm because the base material Al ions were detected. On the other hand, the result of comparing each carbon thickness case suggests that the surface carbon layer is not contributed to generate plasma.

  18. [Estrogen dependance of the progesterone "receptor" in human tissues (author's transl)].

    Science.gov (United States)

    Kreitmann, B; Moure, C; Bayard, F

    1979-01-01

    The concentration of specific progesterone receptor (RP) has been measured in the endometrium of 31 normal menstruating women during the two phases of their menstrual cycle, and compared with the plasma oestradiol-17 beta (E2) concentrations. There was no relationship between RP and plasma E2 when the two phases of the cycle were considered, however a correlation was observed when the follicular phase was only considered. The relationship between oestrogen and RP was then more directly studied in a breast cancer cell line in tissue culture (MCF-7). RP induction by E2 as well as oestriol was demonstrated, but oestrone was ineffective.

  19. Dioestrous ovariectomy: a model to study the role of progesterone in the onset of canine pseudopregnancy.

    Science.gov (United States)

    Gobello, C; Baschar, H; Castex, G; de la Sota, R L; Goya, R G

    2001-01-01

    It has been suggested that overt pseudopregnancy in bitches is caused by an increase in the concentration of serum prolactin as a result of an abrupt decrease in progesterone concentration in the late luteal phase. This hypothesis was tested by using ovariectomy at dioestrus as an experimental model. A total of 18 intact cross- and purebred bitches were used. Eleven animals were ovariectomized (day 0) between day 25 and day 40 of the oestrous cycle, and seven intact bitches were used as controls. Blood samples for determination of prolactin and progesterone concentrations were collected on days -1, 1, 2, 3 and 7 in the ovariectomized group, and on day 1 and day 7 in the control group. On day 7, the presence or absence of overt pseudopregnancy was recorded. The four ovariectomized bitches with a history of pseudopregnancy showed signs of overt pseudopregnancy (P < 0.01). On day 7, progesterone concentrations were significantly higher in the control than in the ovariectomized bitches (P < 0.01). The expected decrease in serum progesterone concentration after ovariectomy was similar in pseudopregnant bitches and non-pseudopregnant bitches. However, in pseudopregnant bitches, but not in non-pseudopregnant bitches, there was a marked increase (expressed as percentage change) in the concentration of prolactin between day -1 and day 7 (P < 0.01). It was concluded that the abrupt decrease in progesterone concentrations does not lead systematically to pseudopregnancy. Only in bitches predisposed to pseudopregnancy would an abrupt decrease in progesterone concentrations induce a substantial increase in prolactin concentrations, which in turn would trigger the typical signs of pseudopregnancy.

  20. [Plasma ibuprofen enantiomers and their pharmacokinetics in Beagle dogs determined by HPLC].

    Science.gov (United States)

    Wang, Hong-yan; Kong, Ai-ying; Yang, Bo; Yan, Liang-ping; Di, Xin

    2015-12-01

    A chiral high-performance liquid chromatography method was developed for the simultaneous determination of ibuprofen enantiomers in dog plasma. It was used to study the pharmacokinetics in the Beagle dog after intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen. Ketoprofen was chosen as the internal standard. After a simple precipitation using methanol as the precipitating solvent, both analytes and IS were separated on a Kromasil 100-5CHI-TBB chiral column (250 mm x4.6 mm, 5 μm) with isocratic elution using acetonitrile - 20 mmol x L(-1) phosphate buffer (pH 3.0, containing 5% methanol) (6 : 4) as the mobile phase. The detection wavelength was 220 nm. Liner calibration curves for both of the ibuprofen enantiomers were over the concentration range from 0.5 to 50 μg x mL(-1) with a lower limit of quantification of 0.5 μg x mL(-1), the accuracies were all in standard ranges. The intra- and inter- assay precisions were all below 7%. The recovery rate was 93.1% to 100.4%. The experiments proved that the method was simple, rapid and sensitive. It can be used in the quantitative determination of ibuprofen enantiomers in dog plasma. The method was used to determine the concentration of ibuprofen enantiomers in Beagle dog plasma after a single intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen (9 mg x kg(-1)) and the pharmacokinetics parameters were calculated based on the concentration-time curves. The C(max) of S-ibuprofen in Beagle dog plasma after a single intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen were 30.8 ± 4.7, 46.1 ± 5.9 and 20.0 ± 2.6 μg x mL(-1), respectively. In terms of the exposure of active ingredient, it revealed a significant difference between the administration of S-ibuprofen and the other two groups. The systematical R- to S- chiral inversion was discussed. Comparing the pharmacokinetic parameters at different doses, chiral inversion were 70.1% ± 36.6% and 76

  1. Progesterone and reproduction in marsupials: a review.

    Science.gov (United States)

    Bradshaw, Felicity J; Bradshaw, Don

    2011-01-01

    Progesterone (P4) profiles throughout pregnancy and the oestrous cycle are reviewed in a wide range of marsupial species, representing 12 Families, and focus on the corpus luteum (CL) and its functioning, compared with its eutherian counterpart. Physiologically, P4 subtends the same fundamental processes supporting gestation in marsupials as it does in eutherian mammals, from its role in stimulating the secretory endometrium, effecting nutritional transfer across the placenta and establishing lactogenesis. Before the formation of the CL, however, secretion of P4 is widespread throughout many Families and the dual roles of P4 in the induction of sexual behaviour and ovulation are exposed. In Dasyuridae, raised levels of P4 are linked with the induction of sexual receptivity and are also present around the time of mating in Burramyidae, Petauridae and Tarsipedidae, but their function is unknown. Only in Didelphidae has research established that the pheromonally-induced levels of pro-oestrous P4 trigger ovulation. This is principally the role of oestradiol in the eutherian and may be an important difference between the marsupial and the eutherian. The deposition of the shell coat around the early marsupial embryo is also a function of P4, but perhaps the most striking difference is seen in the time taken to form the CL. This is not always immediate and the maximum secretion of P4 from the granulosa cells may not occur until some 2 weeks after ovulation. The slower development of the CL in some species is linked with delays in the development of the embryo during its unattached phase and results in relatively long gestation periods. A common feature of these, in monovular species, is a short pulse of P4 from the newly-luteinised CL, which is all that is needed for the subsequent development of the embryo to term. Maternal recognition of pregnancy occurs soon after the formation of the blastocyst, with embryo-induced changes in ovarian production of P4 and the uterine

  2. Estrogen, Progesterone and Epithelial Ovarian Cancer

    Directory of Open Access Journals (Sweden)

    Ho Shuk-Mei

    2003-10-01

    Full Text Available Abstract Ovarian carcinoma (OCa continues to be the leading cause of death due to gynecologic malignancies and the vast majority of OCa is derived from the ovarian surface epithelium (OSE and its cystic derivatives. Epidemiological evidence strongly suggests that steroid hormones, primarily estrogens and progesterone, are implicated in ovarian carcinogenesis. However, it has proved difficult to fully understand their mechanisms of action on the tumorigenic process. New convincing data have indicated that estrogens favor neoplastic transformation of the OSE while progesterone offers protection against OCa development. Specifically, estrogens, particularly those present in ovulatory follicles, are both genotoxic and mitogenic to OSE cells. In contrast, pregnancy-equivalent levels progesterone are highly effective as apoptosis inducers for OSE and OCa cells. In this regard, high-dose progestin may exert an exfoliation effect and rid an aged OSE of pre-malignant cells. A limited number of clinical studies has demonstrated efficacies of antiestrogens, aromatase inhibitors, and progestins alone or in combination with chemotherapeutic drugs in the treatment of OCa. As a result of increased life expectancy in most countries, the number of women taking hormone replacement therapies (HRT continues to grow. Thus, knowledge of the mechanism of action of steroid hormones on the OSE and OCa is of paramount significance to HRT risk assessment and to the development of novel therapies for the prevention and treatment of OCa.

  3. Determination of plasma lactic acid concentration and specific activity using high-performance liquid chromatography.

    Science.gov (United States)

    Bleiberg, B; Steinberg, J J; Katz, S D; Wexler, J; LeJemtel, T

    1991-08-23

    Assessment of lactate metabolism is of particular interest during exercise and in disease states such as diabetes, shock, and absorptive abnormalities of short-chain fatty acids by the colon. We describe an analytical method that introduces radio-active tracers and high-performance liquid chromatography (HPLC) to simultaneously analyze concentrations and specific activities (SAs) of plasma lactate. The HPLC conditions included separation on a reversed-phase column (octadecylsilane) and an isocratic buffer (30% acetonitrile in water). [3H]Acetate served as an internal standard. Lactate and acetate were extracted from plasma samples with diethyl ether following a pH adjustment to less than 1.0 and back-extracted into a hydrophilic phase with sodium carbonate (2 mM, pH greater than 10.0). Lactate is detected in the ultraviolet range (242 and 320 nm) by derivatization with alpha-bromoacetophenone. Control plasma samples were studied after an overnight fast for precision and analytical recovery. Calibration curves were linear in the range 0.18-6.0 mM (r = 0.92). The precision was 3% and the analytical recovery was 87%. The detection limit of the method was 36 pmol. Determination of lactate metabolism was performed in a patient with chronic congestive heart failure who was administered primed-continuous L-[U-14C]lactate (10 microCi bolus and 0.3 microCi/min continuously) during a 60-min rest period. Mean arterial lactate concentration and SA were 1.69 +/- 0.2 mM and 253.8 +/- 22 dpm/mumol, respectively. Systemic lactate turnover was 25.65 mumol/kg per min. Lactic acid systemic turnover, organ uptake and release rates can be accurately determined by isocratic HPLC.

  4. [Determination of minor elements in stainless steel by laser-induced plasma spectroscopy (LIPS)].

    Science.gov (United States)

    Li, Jing; Zhai, Chao; Zhang, Shi-Ding; Zhang, Jian-Qiu; Meng, Xiang-Ru

    2008-04-01

    Laser-induced plasma spectroscopy (LIPS) is characterized by its non-contact and real-time analysis. Its application to the determination of steel composition can meet the need of high-speed, continuous and automatic production in large steel companies. In the present article the minor elements concentrations of aluminum, manganese, cobalt, molybdenum, and titanium in a series of stainless steel 1Cr18Ni9Ti samples were determinate by laser-induced plasma spectroscopy, based on a Nd : YAG Q-switched solid laser with wavelength 1 064 nm as an exciting source and ICCD as detector. In the experiment the working delay time and gate time of ICCD were set suitably to get high signal-to-noise ratio emission spectral lines, and the internal standardization method related to matrix effect was used to deal with spectral data. Experiment results show that the concentration ratios of all the measured elements versus the reference element ferrum have a good linear relationship with the intensity ratios of them, the detection limits of the five tested elements are within 150 microg x g(-1).

  5. "Determination of mycophenolic acid in human plasma by high-performance liquid chromatography"

    Directory of Open Access Journals (Sweden)

    "Mehdi Ahadi Barzoki

    2005-05-01

    Full Text Available A simple, sensitive and reproducible HPLC method is presented for determination of mycophenolic acid(MPA in human plasma. Samples were prepared after precipitation of the plasma protein by addition of acetonitrile and naproxen was used as internal standard (I.S.. Separation was performed by reversedphase HPLC, using a Hamilton PRP-C18 Column, 51% acetonitrile and 49% potassium phosphate buffer (20 mM at pH 3.0 as mobile phase, flow rate of 1.0 ml/min, and UV detection at 215 nm. MPA and I.S. had retention times of 7.5 and 11.35 min, respectively. The method showed an acceptable linearity in the range of 0.1µg/ml-40µg/ml with r2 of .9992. The concentration of 0.1µg/ml was determined as quantification limit. Mean absolute recovery was 94.8%. The mean intra- and inter-day reproducibility of method was 4.6 and 11.4% respectively.

  6. Is a deuterated internal standard appropriate for the reliable determination of olmesartan in human plasma?

    Science.gov (United States)

    Piórkowska, Edyta; Musijowski, Jacek; Buś-Kwaśnik, Katarzyna; Rudzki, Piotr J

    2017-01-01

    A right choice of the internal standard is one of the most challenging tasks during bioanalytical method development. Surprisingly, among the HPLC-MS methods for the determination of a cardiovascular drug olmesartan in plasma only structural analogues or similar compounds were used as internal standards. We have tried to answer the question whether the stable isotope labelled (deuterated) internal standard, as recommended by regulatory agencies, can be used for the reliable determination of olmesartan in human plasma. An HPLC-MS method using this standard in a simplified liquid-liquid extraction procedure led to accurate and precise results in the linearity range of 5-2500ng/mL. The method is well suited for pharmacokinetic studies following a single 40mg oral dose of olmesartan medoxomil in humans. The method was fully validated according to international guidelines and successfully applied in a bioequivalence study in humans. The use of deuterated olmesartan as the internal standard afforded a reliable tool for regulatory bioanalysis that can indirectly contribute to therapy efficacy and improve the safety of patients treated with generic medicines. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Determination of triamterene in human plasma and urine after its cloud point extraction

    Directory of Open Access Journals (Sweden)

    Ahad Bavili Tabrizi

    2014-01-01

    Full Text Available A new analytical approach was developed involving cloud point extraction (CPE and spectrofluorimetric determination of triamterene (TM in biological fluids. A urine or plasma sample was prepared and adjusted to pH 7, then TM was quickly extracted using CPE, using 0.05% (w/v of Triton X-114 as the extractant. The main factors that affected the extraction efficiency (the pH of the sample, the Triton X-114 concentration, the addition of salt, the extraction time and temperature, and the centrifugation time and speed were studied and optimized. The method gave calibration curves for TM with good linearities and correlation coefficients (r higher than 0.99. The method showed good precision and accuracy, with intra- and inter-assay precisions of less than 8.50% at all concentrations. Standard addition recovery tests were carried out, and the recoveries ranged from 94.7% to 114%. The limits of detection and quantification were 3.90 and 11.7 µg L-1, respectively, for urine and 5.80 and 18.0 µg L-1, respectively, for plasma. The newly developed, environmentally friendly method was successfully used to extract and determine TM in human urine samples.

  8. Instability-Enhanced Collisional Friction Determines the Bohm Criterion in Multiple-Ion-Species Plasmas

    Science.gov (United States)

    Baalrud, S. D.; Hegna, C. C.; Callen, J. D.

    2009-11-01

    Ion-ion streaming instabilities are excited in the presheath region of plasmas with multiple ion species if the ions are much colder than the electrons. Streaming instabilities onset when the relative fluid flow between ion species exceeds a critical speed, δVc, of order the ion thermal speeds. Using a generalized Lenard-Balescu theory that accounts for instability-enhanced collective responses [1], one is able to show the instabilities rapidly enhance the collisional friction between ion species far beyond the contribution from Coulomb collisions alone. This strong frictional force determines the relative fluid speed between species. When this condition is combined with the Bohm criterion generalized for multiple ion species, the fluid speed of each ion species is determined at the sheath edge. For each species, this speed differs from the common ``system'' sound speed by a factor that depends on the species concentrations, masses and δVc.[4pt] [1] S.D. Baalrud, J.D. Callen, and C.C. Hegna, Phys. Plasmas 15, 092111 (2008).

  9. Determinants of folate and vitamin B12 plasma levels in the French E3N-EPIC cohort.

    Science.gov (United States)

    de Batlle, Jordi; Matejcic, Marco; Chajes, Veronique; Moreno-Macias, Hortensia; Amadou, Amina; Slimani, Nadia; Cox, David G; Clavel-Chapelon, Françoise; Fagherazzi, Guy; Romieu, Isabelle

    2016-12-21

    Impaired B vitamin status has been identified as a risk factor for major chronic diseases. This study aims at examining the determinants of plasma folate and vitamin B12 concentrations, considering lifestyle factors and MTHFR polymorphisms. A total of 988 women aged 40-65 years from the French E3N cohort were investigated. Intakes of folate and vitamin B12 were assessed using food frequency questionnaires, and plasma concentrations were measured by microbiological assay. Dietary scores were computed to summarize folate and vitamin B12 dietary sources. MTHFR-C677T and MTHFR-A1298C were determined by Kaspar assay. Pearson's partial correlation coefficients and multivariable linear regression models were used to assess correlations between main determinants and plasma folate and vitamin B12 levels. The partial correlation coefficient between dietary intakes and plasma folate was 0.19 (p value vitamin B12. Dietary scores were the main determinant of B vitamin plasma concentrations with a percent change per unit increase of 12.64% (p value vitamin B12. Homozygous (T/T) or heterozygous (C/T) women for MTHFR-C677T had lower plasma folate concentrations [C/T: -6.48% (p value = 0.038) and T/T: -15.89% (p value vitamin plasma concentration include: smoking status for folate, and age and hormone replacement therapy for vitamin B12. We confirmed previous findings on the role of diet as main determinant of folate and vitamin B12 plasma concentrations. However, the impact of genetic polymorphisms and lifestyle factors on plasma B vitamin concentrations should not be neglected.

  10. Determination of {sup 233}U by inductively coupled argon plasma - atomic emission spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Patwardhan, A.B.; Kulkarni, V.T.; Radhakrishnan, K.; Ramanujam, A.; Page, A.G. [Bhabha Atomic Research Centre, Bombay (India)

    1994-09-01

    The paper describes studies carried out for the determination of {sup 233}U at various stages during the recovery and purification of {sup 233}U from {sup 233}U-Al alloy scraps generated during the fabrication of {sup 233}U-Al alloy fuel. Employing a high resolution sequential spectrometer and Inductively Coupled argon Plasma (ICP) as the spectral excitation source, isotope shift for {sup 233}U with respect to {sup 238}U has been resolved and recorded. The shift for the 424.437 nm emission line of {sup 238}U is found to be of the order of 0.040 nm on the lower wavelength side for {sup 233}U and this isotopic effect has been utilised for the quantitative determination of {sup 233}U. The overall precision of the method is 5% RSD with the detection limit of 0.01 {mu}g/ml.

  11. Determination of metals in composite diet samples by inductively coupled plasma-mass spectrometry.

    Science.gov (United States)

    Melnyk, Lisa Jo; Morgan, Jeffrey N; Fernando, Reshan; Pellizzari, Edo D; Akinbo, Olujide

    2003-01-01

    A study was conducted to evaluate the applicability of inductively coupled plasma-mass spectrometry (ICP-MS) techniques for determination of metals in composite diets. Aluminum, cadmium, chromium, copper, lead, manganese, nickel, vanadium, and zinc were determined by this method. Atmospheric pressure microwave digestion was used to solubilize analytes in homogenized composite diet samples, and this procedure was followed by ICP-MS analysis. Recovery of certified elements from standard reference materials ranged from 92 to 119% with relative standard deviations (RSDs) of 0.4-1.9%. Recovery of elements from fortified composite diet samples ranged from 75 to 129% with RSDs of 0-11.3%. Limits of detection ranged from 1 to 1700 ng/g; high values were due to significant amounts of certain elements naturally present in composite diets. Results of this study demonstrate that low-resolution quadrupole-based ICP-MS provides precise and accurate measurements of the elements tested in composite diet samples.

  12. Determination of trace multi-elements in coal fly ash by inductively coupled plasma mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    XIE Hua-lin; TANG You-gen; LI Yu-jie; LI Li-bo

    2007-01-01

    The contents of Cr,Cu, Ni, As, Cd and Pb in coal fly ash were determined by a high resolution inductively coupled plasma mass spectrometry method.The sample digestions were performed in closed microwave vessels with HN03, HClO4 and HF.The optimum conditions for the determination were obtained.The applicability of the proposed method was validated by the analysis of coal fly ash reference material (NIST SRM 1633a). The results show that most of the spectral interferences can be avoided by measuring in the high resolution mode(maximum mass resolution R=9 000).The detection limit is from 0.05 to 0.21μg/g,and the precision is fine with relative standard deviation less than 4.3%.

  13. Progesterone - new therapy in mild carpal tunnel syndrome? Study design of a randomized clinical trial for local therapy

    Directory of Open Access Journals (Sweden)

    Ginanneschi Federica

    2010-04-01

    Full Text Available Abstract Background Local corticosteroid injection for carpal tunnel syndrome (CTS provides greater clinical improvement in symptoms one month after injection compared to placebo but significant symptom relief beyond one month has not been demonstrated and the relapse of symptoms is possible. Neuroprotection and myelin repair actions of the progesterone was demonstrated in vivo and in vitro study. We report the design of a randomized controlled trial for the local injection of cortisone versus progesterone in "mild" idiopathic CTS. Methods Sixty women with age between 18 and 60 years affected by "mild" idiopathic CTS, diagnosed on the basis of clinical and electrodiagnostic tests, will be enrolled in one centre. The clinical, electrophysiological and ultasonographic findings of the patients will be evaluate at baseline, 1, 6 and 12 months after injection. The major outcome of this study is to determine whether locally-injected progesterone may be more beneficial than cortisone in CTS at clinical levels, tested with symptoms severity self-administered Boston Questionnaire and with visual analogue pain scale. Secondary outcome measures are: duration of experimental therapy; improvement of electrodiagnostic and ultrasonographic anomalies at various follow-up; comparison of the beneficial and harmful effects of the cortisone versus progesterone. Conclusion We have designed a randomized controlled study to show the clinical effectiveness of local progesterone in the most frequent human focal peripheral mononeuropathy and to demonstrate the neuroprotective effects of the progesterone at the level of the peripheral nervous system in humans.

  14. Does progesterone show neuroprotective effects on traumatic brain injury through increasing phosphorylation of Akt in the hippocampus?

    Institute of Scientific and Technical Information of China (English)

    Richard Justin Garling; Lora Talley Watts; Shane Sprague; Lauren Fletcher; David F Jimenez; Murat Digicaylioglu

    2014-01-01

    There are currently no federally approved neuroprotective agents to treat traumatic brain injury. Progesterone, a hydrophobic steroid hormone, has been shown in recent studies to exhibit neu-roprotective effects in controlled cortical impact rat models. Akt is a protein kinase known to play a role in cell signaling pathways that reduce edema, inlfammation, apoptosis, and promote cell growth in the brain. This study aims to determine if progesterone modulates the phosphor-ylation of Aktvia its threonine 308 phosphorylation site. Phosphorylation at the threonine 308 site is one of several sites responsible for activating Akt and enabling the protein kinase to carry out its neuroprotective effects. To assess the effects of progesterone on Akt phosphorylation, C57BL/6 mice were treated with progesterone (8 mg/kg) at 1 (intraperitonally), 6, 24, and 48 hours (subcutaneously) post closed-skull traumatic brain injury. The hippocampus was harvest-ed at 72 hours post injury and prepared for western blot analysis. Traumatic brain injury caused a signiifcant decrease in Akt phosphorylation compared to sham operation. However, mice treat-ed with progesterone following traumatic brain injury had an increase in phosphorylation of Akt compared to traumatic brain injury vehicle. Our ifndings suggest that progesterone is a viable treatment option for activating neuroprotective pathways after traumatic brain injury.

  15. Determining the Concentrations and Temperatures of Products in a CF_4/CHF_3/N_2 Plasma via Submillimeter Absorption Spectroscopy

    Science.gov (United States)

    Helal, Yaser H.; Neese, Christopher F.; De Lucia, Frank C.; Ewing, Paul R.; Agarwal, Ankur; Craver, Barry; Stout, Phillip J.; Armacost, Michael D.

    2017-06-01

    Plasmas used for the manufacturing of semiconductor devices are similar in pressure and temperature to those used in the laboratory for the study of astrophysical species in the submillimeter (SMM) spectral region. The methods and technology developed in the SMM for these laboratory studies are directly applicable for diagnostic measurements in the semiconductor manufacturing industry. Many of the molecular neutrals, radicals, and ions present in processing plasmas have been studied and their spectra have been cataloged or are in the literature. In this work, a continuous wave, intensity calibrated SMM absorption spectrometer was developed as a remote sensor of gas and plasma species. A major advantage of intensity calibrated rotational absorption spectroscopy is its ability to determine absolute concentrations and temperatures of plasma species from first principles without altering the plasma environment. An important part of this work was the design of the optical components which couple 500-750 GHz radiation through a commercial inductively coupled plasma chamber. The measurement of transmission spectra was simultaneously fit for background and absorption signal. The measured absorption was used to calculate absolute densities and temperatures of polar species. Measurements for CHF_3, CF_2, FCN, HCN, and CN made in a CF_4/CHF_3/N_2 plasma will be presented. Temperature equilibrium among species will be shown and the common temperature is leveraged to obtain accurate density measurements for simultaneously observed species. The densities and temperatures of plasma species are studied as a function of plasma parameters, including flow rate, pressure, and discharge power.

  16. Inhibition of progesterone metabolism mimics the effect of progesterone withdrawal on forced swim test immobility.

    Science.gov (United States)

    Beckley, Ethan H; Finn, Deborah A

    2007-10-01

    Withdrawal from high levels of progesterone in rodents has been proposed as a model for premenstrual syndrome or postpartum depression. Forced swim test (FST) immobility, used to model depression, was assessed in intact female DBA/2J mice following progesterone withdrawal (PWD) or treatment with the 5alpha-reductase inhibitor finasteride. Following 5 daily progesterone injections (5 mg/kg IP) FST immobility increased only in mice withdrawn for 3 days (pimmobility. PWD and finasteride treatment, both of which reduce allopregnanolone levels, were associated with increased FST immobility in female DBA/2J mice. These findings suggest that decreased levels of the GABAergic neurosteroid allopregnanolone contribute to symptoms of PWD. Future studies of PWD may provide information about human conditions that are associated with hormone changes such as premenstrual syndrome or postpartum depression.

  17. Evaluation of models to induce low progesterone during the early luteal phase in cattle.

    Science.gov (United States)

    Beltman, M E; Roche, J F; Lonergan, P; Forde, N; Crowe, M A

    2009-10-15

    Two experiments were designed to evaluate models for generation of low circulating progesterone concentrations during early pregnancy in cattle. In Experiment 1, 17 crossbred heifers (Bos taurus) were assigned to either prostaglandin F(2alpha) (PGF(2alpha)) administration on Days 3, 3.5, and 4 (PG3; n=9) or to control (n=8). Blood samples were collected from heifers from Days 1 to 9 for progesterone assay. Progesterone concentrations were decreased (P<0.03) between 18 and 48h after first PGF(2alpha) treatment in heifers assigned to PG3 compared with that of controls. In Experiment 2, 39 crossbred heifers detected in estrus were inseminated (Day 0) and assigned to either (1) PGF(2alpha) administration on Days 3, 3.5, and 4 (PG3; n=10), (2) PGF(2alpha) administration on Days 3, 3.5, 4, and 4.5 (PG4; n=10), (3) Progesterone Releasing Intravaginal Device (PRID) insertion on Day 4.5 with PGF(2alpha) administration on Days 5 and 6 (PRID+PGF(2alpha); n=10), or (4) control (n=9). Blood samples were collected daily until Day 15, and conceptus survival rate was determined at slaughter on Day 16. Progesterone concentrations during the sampling period in the PG3 and PG4 groups did not differ but were less than that of controls (P<0.01). After an initial peak, progesterone concentrations in the PRID+PGF(2alpha) group were similar to that of controls. More heifers in the PG4 group (6 of 10) had complete luteal regression than did those in the PG3 group (3 of 10). Conceptus survival rate on Day 16 did not differ between groups. There was a significant correlation between progesterone concentration on Days 5 and 6 and conceptus size on Day 16. In summary, treatment with PGF(2alpha) on Days 3, 3.5, and 4 postestrus appeared to provide the best model to induce reduced circulating progesterone concentrations during the early luteal phase in cattle.

  18. Spectrofluorimetric determination of certain biologically active phenothiazines in commercial dosage forms and human plasma.

    Science.gov (United States)

    Mohamed, Abdel-Maaboud I; Abdelmageed, Osama H; Salem, Hesham; Nagy, Dalia M; Omar, Mahmoud A

    2013-01-01

    A validated simple and sensitive spectrofluorimetric method was developed for the determination of chlorpromazine hydrochloride, promethazine hydrochloride, trifluperazine hydrochloride, thioridazine hydrochloride, perazine maleate and oxomemazine. The method was based on condensation of malonic acid/acetic anhydride (MAA) under the catalytic effect of the tertiary amine moiety of the studied phenothiazines to provide a deep yellow to brown colour with green fluorescence. Relative fluorescence intensity of the products was measured at λ exc 398 nm and λ em 432 nm. Different variables affecting the reaction were studied and optimized. The method was successfully applied for the determination of the studied drugs in commercial dosage forms. The lower detection limits allowed the application of this method for the determination of the compounds in plasma as an example of a biological fluid. In addition, the method was considered specific for the determination of tertiary amines in the presence of primary and secondary amines; as a result, it was deemed suitable for the determination of the cited drugs in the presence of their degradation products resulting from N-dealkylation or oxidation of the corresponding sulphoxides or sulphones.

  19. Progesterone, brain-derived neurotrophic factor and neuroprotection.

    Science.gov (United States)

    Singh, M; Su, C

    2013-06-03

    While the effects of progesterone in the CNS, like those of estrogen, have generally been considered within the context of reproductive function, growing evidence supports its importance in regulating non-reproductive functions including cognition and affect. In addition, progesterone has well-described protective effects against numerous insults in a variety of cell models, animal models and in humans. While ongoing research in several laboratories continues to shed light on the mechanism(s) by which progesterone and its related progestins exert their effects in the CNS, our understanding is still incomplete. Among the key mediators of progesterone's beneficial effects is the family of growth factors called neurotrophins. Here, we review the mechanisms by which progesterone regulates one important member of the neurotrophin family, brain-derived neurotrophic factor (BDNF), and provides support for its pivotal role in the protective program elicited by progesterone in the brain.

  20. Progesterone signals through membrane progesterone receptors (mPRs) in MDA-MB-468 and mPR-transfected MDA-MB-231 breast cancer cells which lack full-length and N-terminally truncated isoforms of the nuclear progesterone receptor

    Science.gov (United States)

    Pang, Yefei; Thomas, Peter

    2011-01-01

    The functional characteristics of membrane progesterone receptors (mPRs) have been investigated using recombinant mPR proteins over-expressed in MDA-MB-231 breast cancer cells. Although these cells do not express the full-length progesterone receptor (PR), it is not known whether they express N-terminally truncated PR isoforms which could possibly account for some progesterone receptor functions attributed to mPRs. In the present study, the presence of N-terminally truncated PR isoforms was investigated in untransfected and mPR-transfected MDA-MB-231 cells, and in MDA-MB-468 breast cancer cells. PCR products were detected in PR-positive T47D Yb breast cancer cells using two sets of C-terminus PR primers, but not in untransfected and mPR-transfected MDA-MB-231 cells, nor in MDA-MB-468 cells. Western blot analysis using a C-terminal PR antibody, 2C11F1, showed the same distribution pattern for PR in these cell lines. Another C-terminal PR antibody, C-19, detected immunoreactive bands in all the cell lines, but also recognized α-actinin, indicating that the antibody is not specific for PR. High affinity progesterone receptor binding was identified on plasma membranes of MDA-MB-468 cells which was significantly decreased after treatment with siRNAs for mPRα and mPRβ. Plasma membranes of MDA-MB-468 cells showed very low binding affinity for the PR agonist, R5020, ≤1% that of progesterone, which is characteristic of mPRs. Progesterone treatment caused G protein activation and decreased production of cAMP in MDA-MB-468 cells, which is also characteristic of mPRs. The results indicate that the progestin receptor functions in these cell lines are mediated through mPRs and do not involve any N-terminally truncated PR isoforms. PMID:21291899

  1. Progesterone receptor B promoter hypermethylation in human placenta after labor onset.

    Science.gov (United States)

    Zhuang, Yanyan; Cui, Hong; Liu, Sishi; Zheng, Dongming; Liu, Caixia

    2015-03-01

    To determine the methylation status of progesterone receptor B (PR-B) promoter and how PR-B regulates progesterone action in placenta during human pregnancy. Placentas were obtained from the pregnancy women at term who underwent cesarean delivery and vaginal delivery. The methylation status of the PR-B promoter was analyzed using the methylation-specific polymerase chain reaction (PCR) and bisulfite sequencing PCR. And the messenger RNA (mRNA) and protein expression of the PR-B and DNA methyltransferases (DNMTs) were determined by quantitative real-time PCR and Western blot. Compared with the cesarean group, the placentas of vaginal delivery group had greater levels of PR-B DNA methylation, and the PR-B, DNMT1, DNMT3a, and DNMT3b mRNA and protein expression were significantly decreased. Progesterone receptor B methylation occurs with high frequency after labor onset and may play an important epigenetic mechanism of labor-related PR-B negative expression, thereby mediating the biological process of functional progesterone withdrawal at term for parturition. © The Author(s) 2014.

  2. Dehydroepiandrosterone (DHEA) supplementation results in supraphysiologic DHEA-S serum levels and progesterone assay interference that may impact clinical management in IVF.

    Science.gov (United States)

    Franasiak, Jason M; Thomas, Semara; Ng, Susan; Fano, Maria; Ruiz, Andrew; Scott, Richard T; Forman, Eric J

    2016-03-01

    Dehydroepiandrosterone (DHEA) is often prescribed for poor responders in IVF in an effort to improve response to ovarian stimulation. The effect of DHEA supplementation and resultant supraphysiologic DHEA-S serum levels on sex steroid assays has not been evaluated in this population. This study seeks to determine the relationship between DHEA supplementation and progesterone measurements to characterize the degree of interference with particular immunoassays. Characterization was accomplished in two phases. First, DHEA-S standard control reagents with no progesterone present were assayed for both DHEA-S and progesterone levels. Second, serum pools from 60 unique IVF patients' serum were used to create six pooled serum samples: three from patients on DHEA supplementation and three from patients not on DHEA supplementation. The three pools were composed of patients whose serum fell into low, medium, and high progesterone ranges. Baseline DHEA-S and progesterone were measured, and the mean level of DHEA-S in the mid-range progesterone pool was used as the mid-point for addition of DHEA-S standard to the serum pools from patients without DHEA supplementation. Progesterone from these pools was then measured on three commercially available immunoassay systems. The first experiment revealed a linear increase in progesterone when analyzing the DHEA-S standard ranging from 0.5 ng/mL in the blank control (no DHEA-S) to up to 2.0 ng/mL in the high control (DHEA-S >700 μg/mL), indicating that the DHEA-S cross-reacts with the progesterone assays. In the second experiment, patients' serum DHEA-S and progesterone were measured from pooled serum samples of those taking DHEA and those not taking DHEA. Adding DHEA-S to the pooled serum of those not taking DHEA resulted in a linear increase in progesterone levels on two of three commercially available immunoassays (p DHEA-S can interfere with standard progesterone immunoassays used in clinical ART programs, and thus serum

  3. Determination of sodium cromoglycate in human plasma by liquid chromatography with tandem mass.

    Science.gov (United States)

    Liu, Xiao-yan; Qu, Ting-ting; Wang, Ben-jie; Wei, Chun-min; Yuan, Gui-yan; Zhang, Rui; Guo, Rui-chen

    2008-09-01

    A sensitive and selective liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the determination of sodium cromoglycate (SCG) in human plasma after a nasal dose of 10.4 mg sodium cromoglycate nasal spray, using pravastatin sodium as the internal standard. The method was validated over a linear range of 0.300-20.0 ng/mL. SCG and I.S. were extracted from 1.0 mL of heparinized plasma by C(18) solid-phase extraction cartridges using methanol as eluting solvent. The dried residue was reconstituted with 100 microL of mobile phase, and 10 microL was injected onto the LC-MS/MS system. Chromatographic separation was achieved on a C(18) column (250 x 4.6 mm i.d., 5 microm particle size) with a mobile phase of methanol-acetonitrile-water (containing 2 mmol/L ammonium acetate; 42.5:42.5:15, v/v/v) at a flow rate of 0.4 mL/min. The analytes were detected with a triple quad LC-MS/MS using ESI with positive ionization. Ions monitored in the multiple reaction monitoring mode were m/z 469.0 (precursor ion) to m/z 245.0 (product ion) for SCG and m/z 447.2 (precursor ion) to m/z327.1 (product ion) for pravastatin sodium (internal standard) The average recovery of SCG from human plasma was 94.88% and the lower limit of quantitation was 0.3 ng/mL. Results from a 3-day validation study demonstrated excellent precision and accuracy across the calibration range of 0.3-20 ng/mL. The method was successfully applied to the pharmacokinetic study of SCG in healthy Chinese volunteers.

  4. High Performance Liquid Chromatographic Assay for the Simultaneous Determination of Posaconazole and Vincristine in Rat Plasma

    Directory of Open Access Journals (Sweden)

    Hadeel A. Khalil

    2015-01-01

    Full Text Available Purpose. Developing a validated HPLC-DAD method for simultaneous determination of posaconazole (PSZ and vincristine (VCR in rat plasma. Methods. PSZ, VCR, and itraconazole (ITZ were extracted from 200 μL plasma using diethyl ether in the presence of 0.1 M sodium hydroxide solution. The organic layer was evaporated in vacuo and dried residue was reconstituted and injected through HC-C18 (4.6 × 250 mm, 5 μm column. In the mobile phase, acetonitrile and 0.015 M potassium dihydrogen orthophosphate (30 : 70 to 80 : 20, linear gradient over 7 minutes pumped at 1.5 mL/min. VCR and PSZ were measured at 220 and 262 nm, respectively. Two Sprague Dawley rats were orally dosed PSZ followed by iv dosing of VCR and serial blood sampling was performed. Results. VCR, PSZ, and ITZ were successfully separated within 11 min. Calibration curves were linear over the range of 50–5000 ng/mL for both drugs. The CV% and % error of the mean were ≤18% and limit of quantitation was 50 ng/mL for both drugs. Rat plasma concentrations of PSZ and VCR were simultaneously measured up to 72 h and their calculated pharmacokinetics parameters were comparable to the literature. Conclusion. The assay was validated as per ICH guidelines and is appropriate for pharmacokinetics drug-drug interaction studies.

  5. Validated enzyme-linked immunosorbent assay for determination of rosuvastatin in plasma at picogram level.

    Science.gov (United States)

    Darwish, Ibrahim A; Al-Obaid, Abdul-Rahman M; Al-Malaq, Hamoud A

    2013-05-01

    In this study, a highly sensitive enzyme-linked immunosorbent assay (ELISA) has been developed and validated for the determination of rosuvastatin (ROS) in plasma samples at picogram level. The assay employed a polyclonal antibody that specifically recognizes ROS with high affinity, and ROS conjugate of bovine serum albumin (ROS-BSA) immobilized onto microplate wells as a solid phase. The assay involved a competitive binding reaction between ROS, in plasma sample, and the immobilized ROS-BSA for the binding sites on a limited amount of the anti-ROS antibody. The bound anti-ROS antibody was quantified with horseradish peroxidase-labelled second anti-rabbit IgG antibody (HRP-IgG) and 3,3`,5,5`-tetramethylbenzidine (TMB) as a substrate for the peroxidase enzyme. The concentration of ROS in the sample was quantified by its ability to inhibit the binding of the anti-ROS antibody to the immobilized ROS-BSA and subsequently the colour intensity in the assay wells. The assay limit of detection was 25 pg ml(-1) and the effective working range at relative standard deviations (RSD) of ≤ 5% was 40-2000 pg ml(-1). Analytical recovery of ROS from spiked plasma was 96.2 - 104.8 ± 2.12 - 5.42%. The precision of the assay was satisfactory; RSD was 2.47 - 4.46 and 3.24 - 5.27% for the intra- and inter-assay precision, respectively. The analytical procedure is convenient, and one can analyze ~ 200 samples per working day, facilitating the processing of large-number batch of samples. The proposed ELISA has a great value in routine analysis of ROS for its pharmacokinetic studies.

  6. Determination of ammonium and organic bound nitrogen by inductively coupled plasma emission spectroscopy.

    Science.gov (United States)

    Jaber, A M Y; Mehanna, N A; Sultan, S M

    2009-06-15

    The continuous flow sample introduction technique with a hydride generator system in conjunction with an inductively coupled plasma emission spectrometer (ICP-AES-HG), is used in this study for quantitative determination of ammonium and organic bound nitrogen in aqueous and solid samples. Ammonia vapor released from ammonium salt after treatment with concentrated NaOH is transferred by argon to plasma for detection at 174.273 nm using axial argon plasma mode. The calibration curves were linear within a range of 25-1000 mg L(-1)N as ammonium molybdate with correlation coefficients of better than 0.99 and limits of detection of about 10-25mg L(-1)N. The percent recovery of N (25-500 mg L(-1)N) in soft (distilled) water and high salt content (1.7 mol L(-1) NaCl) matrices was found to be in the range of about 97-102% with %RSD in the range of 4.6-0.62. The sensitivity, limit of detection, and blank contribution from the atmospheric nitrogen, were tremendously improved in this method compared with the available ICP-AES spray chamber counterpart. Furthermore, the ICP-AES-HG method gave results for real samples (soil, fertilizer, waste water) containing about 50-1800 mg L(-1)N in good agreement with those obtained by the standard Kjeldahl method. No statistical differences at the 95% confidence level on applying the t-test were observed between the values obtained by the two methods. Thus, the ICP-AES-HG method is reliable and faster than the conventional tedious Kjeldahl method, superior to the ICP-AES spray chamber method, and almost free from matrix interference which is usually a critical factor in atomic emission spectroscopic techniques.

  7. Evaluation of an immunoassay for determination of plasma efavirenz concentrations in resource-limited settings

    Directory of Open Access Journals (Sweden)

    Alemseged Abdissa

    2014-06-01

    Full Text Available Introduction: Therapeutic drug monitoring (TDM may improve antiretroviral efficacy through adjustment of individual drug administration. This could result in reduced toxicity, prevent drug resistance, and aid management of drug–drug interactions. However, most measurement methods are too costly to be implemented in resource-limited settings. This study evaluated a commercially available immunoassay for measurement of plasma efavirenz. Methods: The immunoassay-based method was applied to measure efavirenz using a readily available Humastar 80 chemistry analyzer. We compared plasma efavirenz concentrations measured by the immunoassay with liquid chromatography tandem mass spectrometry (LC-MS/MS (reference method in 315 plasma samples collected from HIV patients on treatment. Concentrations were categorized as suboptimal4 µg/ml. Agreement between results of the methods was assessed via Bland-Altman plot and κ statistic values. Results: The median Interquartile range (IQR efavirenz concentration was 2.8 (1.9; 4.5 µg/ml measured by the LC–MS/MS method and 2.5 (1.8; 3.9 µg/ml by the immunoassay and the results were well correlated (ρ=0.94. The limits of agreement assessed by Bland–Altman plots were −2.54; 1.70 µg/ml. Although immunoassay underestimated high concentrations, it had good agreement for classification into low, normal or high concentrations (K=0.74. Conclusions: The immunoassay is a feasible alternative to determine efavirenz in areas with limited resources. The assay provides a reasonable approximation of efavirenz concentration in the majority of samples with a tendency to underestimate high concentrations. Agreement between tests evaluated in this study was clinically satisfactory for identification of low, normal and high efavirenz concentrations.

  8. Determination of triapine, a ribonucleotide reductase inhibitor, in human plasma by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Feng, Ye; Kunos, Charles A; Xu, Yan

    2015-09-01

    Triapine is an inhibitor of ribonucleotide reductase (RNR). Studies have shown that triapine significantly decreases the activity of RNR and enhanced the radiation-mediated cytotoxicity in cervical and colon cancer. In this work, we have developed and validated a selective and sensitive LC-MS/MS method for the determination of triapine in human plasma. In this method, 2-[(3-fluoro-2-pyridinyl)methylene] hydrazinecarbothioamide (NSC 266749) was used as the internal standard (IS); plasma samples were prepared by deproteinization with acetonitrile; tripaine and the IS were separated on a Waters Xbridge Shield RP18 column (3.5 µm; 2.1 × 50 mm) using a mobile phase containing 25.0% methanol and 75.0% ammonium bicarbonate buffer (10.0 mM, pH 8.50; v/v); column eluate was monitored by positive turbo-ionspray tandem mass spectrometry; and quantitation of triapine was carried out in multiple-reaction-monitoring mode. The method developed had a linear calibration range of 0.250-50.0 ng/mL with correlation coefficient of 0.999 for triapine in human plasma. The IS-normalized recovery and the IS-normalized matrix factor of triapine were 101-104% and 0.89-1.05, respectively. The accuracy expressed as percentage error and precision expressed as coefficient of variation were ≤±6 and ≤8%, respectively. The validated LC-MS/MS method was applied to the measurement of triapine in patient samples from a phase I clinical trial.

  9. Liquid Chromatography-Tandem Mass Spectrometric Assay for Determination of Stavudine in Human Plasma

    Directory of Open Access Journals (Sweden)

    Fengdan Jin

    2014-01-01

    Full Text Available A LC-MS/MS method for determination of stavudine in human plasma was established and validated, and it was applied to the pharmaceutical formulations bioequivalence study. 0.5 mL plasma sample was extracted by liquid-liquid extraction. Stavudine was detected by a LC-MS/MS system. The pharmacokinetic parameters of stavudine in different formulations were calculated by noncompartment model statistics. The method was linear over the concentration ranges 5.00–1000 ng/mL in plasma. The intra- and interassay relative standard deviation (RSD was <10%. The average accuracies for the assay at three concentrations (5.00, 80.0, and 900 ng/mL were from 100.2% to 102.5%. Pharmacokinetic parameters of stavudine reference formulation were obtained as follows: Tmax was 0.6±0.2 h, Cmax was 480.7±150.9 g/L, t1/2 was 1.7±0.4 h, and AUC0-t was 872.8±227.8 g·h/L, and pharmacokinetic parameters of stavudine test formulation were obtained as follows: Tmax was 0.5±0.2 h, Cmax was 537.5±178.5 g/L, t1/2 was 1.7±0.3 h, and AUC0-t was (914.1±284.5 g·h/L. Calculated with AUC0-t, the bioavailability of two formulations was 105.0%.

  10. New Spectrofluorimetric Method with Enhanced Sensitivity for Determination of Paroxetine in Dosage Forms and Plasma

    Directory of Open Access Journals (Sweden)

    Ibrahim A. Darwish

    2008-01-01

    Full Text Available New simple spectrofluorimetric method with enhanced sensitivity has been developed and validated for the determination of the antidepressant paroxetine (PXT in its dosage forms and plasma. The method was based on nucleophilic substitution reaction of PXT with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole in an alkaline medium (pH 8 to form a highly fluorescent derivative that was measured at 545 nm after excitation at 490 nm. The factors affecting the reaction was carefully studied and optimized. The kinetics of the reaction was investigated, and the reaction mechanism was presented. Under the optimized conditions, linear relationship with good correlation coefficient (0.9993 was found between the fluorescence intensity and PXT concentration in the range of 80–800 ng ml−1. The limits of detection and quantitation for the method were 25 and 77 ng ml−1, respectively. The precision of the method was satisfactory; the values of relative standard deviations did not exceed 3%. The proposed method was successfully applied to the determination of PXT in its pharmaceutical tablets with good accuracy; the recovery values were 100.2 ± 1.61%. The results obtained by the proposed method were comparable with those obtained by the official method. The proposed method is superior to the previously reported spectrofluorimetric method for determination of PXT in terms of its higher sensitivity and wider linear range. The high sensitivity of the method allowed its successful application to the analysis of PXT in spiked human plasma. The proposed method is practical and valuable for its routine application in quality control and clinical laboratories for analysis of PXT.

  11. The benefits of progesterone therapy in imminent abortion

    Directory of Open Access Journals (Sweden)

    A. Abadi

    2005-12-01

    Full Text Available The causes of imminent abortion are multi-factorial. The biggest causal factor is the low level of serum progesterone level. The lowest critical level of serum progesterone for survivability of pregnancy is 10 ng/ml. Eighty percent of patients experiencing abortion showed that their progesterone level was < 10 ng/ml. Patients who realized that their pregnancy would experience hemorrhage generally would suffer from depression. Stress was one of the factors responsible for the occurence of abortion. Administration of natural progesterone substitution (not  progestogen accelerates the disappearance of uterine contractions, and speeds up the stoppage of bleeding. In addition, progesterone has the effect of anti-anxiety. Adminstration of oral progesterone would result in metabolism in the intestine and liver, such that physiological level of serum progesterone could not be reached, while administration of suppositoria progesterone would result in physiological level of serum, such that it was effective to prevent imminent abortion. (Med J Indones 2005; 14:258-62Keywords: progesterone, imminent abortion

  12. Determination of selenoprotein P in human plasma by solid phase extraction and inductively coupled plasma mass spectrometry

    DEFF Research Database (Denmark)

    Bendahl, L.; Sidenius, U.; Gammelgaard, Bente

    2000-01-01

    measured by inductively coupled plasma mass spectrometry (ICP-MS) monitoring the Se-82 isotope. Linear response was observed in the concentration range 0.3-70.8 mu g/l selenium as selenoprotein P with a correlation coefficient of 0.9994. The precision expressed as relative standard deviation was better...

  13. [Determination of chlorine in gasoline by inductively coupled plasma atomic emission spectrometry].

    Science.gov (United States)

    Zhao, Yan; Chen, Xiao-yan; Xu, Dong-yu; Zhang, Shi-yuan; Chen, Ze-yong

    2014-12-01

    A new method was studied for the analysis of chlorine in gasoline by inductively coupled plasma atomic emission spectrometry (ICP-AES). Samples werediluted 1+4(φ) with kerosene. The intense spectral line for chlorine (134.724 nm) was used. In order to eliminate carbon and maintain stable plasma, small amounts of oxygen (0.050 L · min(-1)) were added to the auxiliary gas. The instrumental main condition was optimized in terms of effects of generator power, nebulizer gas flow, auxiliary gas flow, and oxygen flow on SBR for chlorine. Standard addition method was used to compensate matrix effect and signal drift. The recovery for spiking gasoline samples and the limit of detection were in the range of 96.6%~103.9% and 0.27 mg · L(-1) respectively. The relative standard deviation (RSD) was between 1.57% and 4.49%. Compared with microcoulometry, the analysis results of organic chlorine were basically the same. Moreover, chlorine content, including organic chlorine and inorganic chloride was determined by ICP-AES. The proposed method had the advantages of simplicity, speediness and sensitivity, and expanded the ICP-AES application in non-metals especially halogen elements. It can be used for the analysis of chlorine in gasoline and provides technical support for quality evaluation.

  14. Monitoring trace metals in urban aerosols from Buenos Aires city. Determination by plasma-based techniques.

    Science.gov (United States)

    Smichowski, Patricia; Gómez, Dario R; Dawidowski, Laura E; Giné, María Fernanda; Bellato, Ana Claudia Sánchez; Reich, Silvia L

    2004-04-01

    A study was undertaken, within the framework of a 3 years national project, to assess the content of 13 elements in airborne particulate matter collected in representative zones of the metropolitan area of Buenos Aires. The sampling strategy followed consisted in collecting simultaneously 67 samples of PM10 particulate matter in 9 sampling sites covering an area of about 30 km2 during one week. The collection was performed on ash-free fibre-glass filters using high volume samplers. A combination of aqua regia and perchloric acid was used for leaching metals from filters. Key elements, namely Al, Ca, Cu, Fe, Mn, Mo, Ni, Pb, S, Sb, Sn, Zn and Zr, were determined by inductively coupled plasma-optical emission spectrometry (ICP-OES) and inductively coupled plasma-mass spectrometry (ICP-MS) at micro g g(-1) and ng g(-1) levels. Analyte concentration varied from 130 ng g(-1)(Mo) to over 30%(Ca). Multivariate statistical analysis was performed on the data set including the measured elemental compositions for the monitored period. The atmospheric concentration found for Pb confirms the decreasing levels of this element since the introduction of unleaded gasoline in 1995: 88 ng m(-3)(2001) gas imply low emissions of this element from combustion activities. To the best of our knowledge, S concentrations are reported for the first time for this city.

  15. Determination of strychnine and brucine in rat plasma using liquid chromatography electrospray ionization mass spectrometry.

    Science.gov (United States)

    Xu, Yanyan; Si, Duanyun; Liu, Changxiao

    2009-02-20

    A simple, sensitive and selective liquid chromatography-electrospray mass spectrometric (LC-ESI-MS) method was developed and validated for simultaneous determination of strychnine and brucine in rat plasma, using tacrine as the internal standard (IS). Sample preparation involved a liquid-liquid extraction of the analytes with n-hexane, dichloromethane and isopropanol (65:30:5, v/v/v) from 0.1mL of plasma. Chromatographic separation was carried out on a Waters C(18) column using a mobile phase of methanol-20mM ammonium formate-formic acid (32:68:0.68, v/v/v). Positive selected ion monitoring mode was used for detection of strychnine, brucine and the IS at m/z 335.2, m/z 395.2 and m/z 199.2, respectively. Linearity was obtained over the concentration range of 0.5-500ng/mL for strychnine and 0.1-100ng/mL for brucine. The lower limit of quantification was 0.5ng/mL and 0.1ng/mL for strychnine and brucine, respectively. The intra- and inter-day precision for both strychnine and brucine was less than 7.74%, and accuracy ranged from -4.38% to 2.21% at all QC levels. The method has been successfully applied to a pharmacokinetic study of processed Semen Strychni after oral administration to rats.

  16. Determination of piroxicam from rat articular tissue and plasma based on LC-MS/MS.

    Science.gov (United States)

    Kim, Han Sol; Cho, Ha Ra; Ho, Myoung Jin; Kang, Myung Joo; Choi, Yong Seok

    2016-12-01

    Osteoarthritis (OA) is the most common type of arthritis. To manage OA, in general, oral administration of non-steroidal anti-inflammatory drugs (NSAIDs) is used. Recently, the analgesic and anti-inflammatory efficacy of piroxicam (PX), a long-acting NSAID, by intra-articular (IA) administration in OA was reported, and the possibility that PX is distributed in articular tissues at a certain concentration was raised. Thus, herein, novel LC-MS/MS methods to detect PX in rat articular tissue and plasma are presented. For articular tissue, solvent extraction with acetonitrile for 12 h was employed and a protein precipitation method was used for the preparation of a plasma sample. The developed methods were validated by following the FDA guidelines, and the validated methods were successfully applied to a PK study of IA PX. The present study presents, to our knowledge, the first method of determining a drug in articular tissue. Additionally, the level of PX in articular tissue after IA PX administration was experimentally confirmed for the first time using the present methods. Therefore, the present methods provide a new direction for in vivo evaluation for IA PX formulations and contribute to the development of alternative IA PX formulations with better effects for the treatment of OA.

  17. Determination of additives in PVC material by UV laser ablation inductively coupled plasma atomic emission spectrometry

    Science.gov (United States)

    Hemmerlin, M.; Mermet, J. M.; Bertucci, M.; Zydowicz, P.

    1997-04-01

    UV laser ablation inductively coupled plasma atomic emission spectrometry (LA-ICP-AES) has been applied to the direct determination of additives in solid poly(vinyl chloride) materials. A Nd:YAG laser, operating at its fourth harmonic (266 nm), was used with a beam masking device, in the most reproducible conditions, to introduce solid particles into the plasma torch of a simultaneous ICP-AES system. Emphasis was placed on both precision and accuracy in the analysis of PVC materials by LA-ICP-AES. A series of six in-house PVC reference materials was prepared by incorporating several additives in increasing concentrations. Three alternative methods were evaluated to certify the amount of incorporated elements: ICP-AES with sample dissolution, NAA and XRF. Satisfactory results and good agreement were obtained for seven elements (Al, Ca, Cd, Mg, Sb, Sn and Ti) among the ten incorporated. Sample homogeneity appeared to be satisfactory, and calibration graphs obtained by LA-ICP-AES for several elements are presented. Finally, the performance of the technique in terms of repeatability (1.6-5%), reproducibility (2-5%), and limits of detection was investigated.

  18. DETERMINATION OF PLASMA LEVELS OF UREA AND CREATININE IN PANTANAL HORSES POCONÉ REGION - MT

    Directory of Open Access Journals (Sweden)

    M. R. Guazina

    2016-09-01

    Full Text Available This study aimed to determine the plasma levels of urea and creatinine, besides evaluating the influence of sex and age factors on these substances in Pantaneiros horses, animals of high economic and social importance in the Pantanal. In Poconé-MT region, 80 animals were used, which were divided into four groups: Group 1: male foals (n= 20, aged 8 months and 2 years, Group 2: female foals (n= 20, aged 8 months and 2 years, Group 3: adult males (n= 20, aged from 3 years, Group 4: adult females (n= 20, aged from 3 years. All animals were classified as healthy after clinical examinations, as well as serum negativity for Equine Infectious Anemia (EIA. Blood samples for subsequent laboratory analysis were taken from the jugular vein. The mean values of urea and creatinine found for Pantaneiros horses were similar to other breeds of horses and did not vary influenced by the sexual factor, however plasma levels of creatinine increased with the age

  19. [Simultaneous determination of gestodene, etonogestrel and ethinylestradiol in plasma by LC-MS/MS following derivatization].

    Science.gov (United States)

    Liu, Xiao-Fen; Ding, Cun-Gang; Ge, Qing-Hua; Zhou, Zhen; Zhi, Xiao-Jin

    2010-01-01

    To establish a sensitive and specific method for simultaneous determination of gestodene, etonogestrel and ethinylestradiol in plasma by LC-MS/MS, plasma samples were extracted and derivatized before injection. An ESI ion source was used and operated in the positive ion mode with multiple reaction monitoring (MRM). Norgestrel was chosen as internal standard and performed on a C18 (100 mm x 2.1 mm, 5 microm) column. The concentrations of gestodene, etonogestrel and ethinylestradiol were measured, using step-gradient mobile phase and step-gradient flow rate. The method was validated over the concentration range of 0.1-20 ng x mL(-1) for gestodene and etonogestrel and 0.01-2 ng x mL(-1) for ethinylestradiol, and showed excellent linearity. The intra- and inter-assay accuracy and precision were below 10.0% and recovery was 93.6%-110.9% over the three concentration levels evaluated. The method was applied in pharmacokinetic study of the compound gestodene patch and the compound etonogestrel patch in rabbits. The LC-MS/MS method was selective, accurate and sensitive, especially the LOQ were 100 pg x mL(-1) for gestodene and etonogestrel and 10 pg x mL(-1) for ethinylestradiol. The method was successfully applied in pharmacokinetic study for contraceptives.

  20. Determination of mycophenolic acid in human plasma by ultra performance liquid chromatography tandem mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    Vivek Upadhyay; Vikas Trivedi; Gaurang Shah; Manish Yadav; Pranav S. Shrivastav

    2014-01-01

    A simple, sensitive and high throughput ultra performance liquid chromatography tandem mass spectrometry method has been developed for the determination of mycophenolic acid in human plasma. The method involved simple protein precipitation of MPA along with its deuterated analog as an internal standard (IS) from 50 mL of human plasma. The chromatographic analysis was done on Acquity UPLC C18 (100mm*2.1mm,1.7mm) column under isocratic conditions using acetonitrile and 10 mM ammonium formate, pH 3.00 (75:25, v/v) as the mobile phase. A triple quadrupole mass spectrometer operating in the positive ionization mode was used for quantitation. In-source conversion of mycophenolic glucuronide metabolite to the parent drug was selectively controlled by suitable optimization of cone voltage, cone gas flow and desolvation temperature. The method was validated over a wide concentration range of 15–15000 ng/mL. The mean extraction recovery for the analyte and IS was 495%. Matrix effect expressed as matrix factors ranged from 0.97 to 1.02. The method was successfully applied to support a bioequivalence study of 500 mg mycophenolate mofetil tablet in 72 healthy subjects.

  1. The self-consistent determination of HF electroconductivity of strongly coupled plasmas

    CERN Document Server

    Sreckovic, V A; Ignjatovic, Lj M; Mihajlov, A A; 10.1016/j.physleta.2009.11.073

    2012-01-01

    Here is presented the calculation of the dynamic electrical conductivity of fully ionized, strongly coupled plasmas as a function of the external electric field frequency $\\omega$. The calculations are based on the the formula for the energy-dependent collision frequency which is determined by means of the Green function theory methods, as a sum over the Matsubara frequencies. The domain of extremely high electron density: $10^{21}\\leq n_{e}\\leq 10^{24} \\textrm{cm}^{-3}$, and for the temperature varying from $10 \\textrm{kK}$ to $1.000 \\textrm{kK}$ was examined. The real and imaginary parts of the conductivity for every electron density are presented in the generalized Drude-like form as a two-parameter function of the frequency $\\omega$ in the region $0 < \\omega < 0.5\\omega_{p}$, where $\\omega_{p}$ is the plasma frequency. A good agreement between the obtained results and the existing theoretical and computing simulation data is shown.

  2. Coronal magnetic field and the plasma beta determined from radio and multiple satellite observations

    CERN Document Server

    Iwai, Kazumasa; Nozawa, Satoshi; Takahashi, Takuya; Sawada, Shinpei; Kitagawa, Jun; Miyawaki, Shun; Kashiwagi, Hirotaka

    2014-01-01

    We derived the coronal magnetic field, plasma density, and temperature from the observation of polarization and intensity of radio thermal free-free emission using the Nobeyama Radioheliograph (NoRH) and extreme ultraviolet (EUV) observations. We observed a post-flare loop on the west limb 11 April 2013. The line-of-sight magnetic field was derived from the circularly polarized free-free emission observed by NoRH. The emission measure and temperature were derived from the Atmospheric Imaging Assembly (AIA) onboard Solar Dynamics Observatory (SDO). The derived temperature was used to estimate the emission measure from the NoRH radio free-free emission observations. The derived density from NoRH was larger than that determined using AIA, which can be explained by the fact that the low temperature plasma is not within the temperature coverage of the AIA filters used in this study. We also discuss the other observation of the post-flare loops by the EUV Imager onboard the Solar Terrestrial Relations Observatory (...

  3. Plasma Membrane Protein Ubiquitylation and Degradation as Determinants of Positional Growth in Plants

    Institute of Scientific and Technical Information of China (English)

    Barbara Korbei; Christian Luschnig

    2013-01-01

    Being sessile organisms, plants evolved an unparalleled plasticity in their post-embryonic development, allowing them to adapt and fine-tune their vital parameters to an ever-changing environment. Cross-talk between plants and their environment requires tight regulation of information exchange at the plasma membrane (PM). Plasma membrane proteins mediate such communication, by sensing variations in nutrient availability, external cues as well as by controlled solute transport across the membrane border. Localiza-tion and steady-state levels are essential for PM protein function and ongoing research identified cis- and trans-acting determinants, involved in control of plant PM protein localization and turnover. In this overview, we summarize recent progress in our understanding of plant PM protein sorting and degradation via ubiquitylation, a post-translational and reversible modification of proteins. We highlight characterized components of the machinery involved in sorting of ubiquitylated PM proteins and discuss consequences of protein ubiquitylation on fate of selected PM proteins. Specifically, we focus on the role of ubiquitylation and PM protein degradation in the regulation of polar auxin transport (PAT). We combine this regulatory circuit with further aspects of PM protein sorting control, to address the interplay of events that might control PAT and polarized growth in higher plants.

  4. Determination and pharmacokinetic studies of arecoline in dog plasma by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Li, Bing; Zhou, Xu-Zheng; Li, Jian-Yong; Yang, Ya-Jun; Niu, Jian-Rong; Wei, Xiao-Juan; Liu, Xi-Wang; Li, Jin-Shan; Zhang, Ji-Yu

    2014-10-15

    A rapid and sensitive high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of arecoline concentration in dog plasma. Plasma sample was prepared by protein precipitation using n-hexane (containing 1% isoamyl alcohol) with β-pinene as an internal standard. Chromatographic separation was achieved on an Agilent C18 column (4.6×75mm, 3.5μm) using methanol: 5mM ammonium acetate as the mobile phase with isocratic elution. Mass detection was carried out using positive electrospray ionization in multiple reaction monitoring mode. The calibration curve for arecoline was linear over a concentration range of 2-500ng/mL. The intra-day and inter-day accuracy and precision were within the acceptable limits of ±10% at all concentrations. In summary, the LC-MS/MS method described herein was fully validated and successfully applied to the pharmacokinetic study of arecoline hydrobromide tablets in dogs after oral administration. Copyright © 2014. Published by Elsevier B.V.

  5. Determination of the neutral oxygen atom density in a plasma reactor loaded with metal samples

    Energy Technology Data Exchange (ETDEWEB)

    Mozetic, Miran; Cvelbar, Uros [Jozef Stefan Institute, Jamova cesta 39, 1000 Ljubljana (Slovenia)], E-mail: miran.mozetic@ijs.si

    2009-08-15

    The density of neutral oxygen atoms was determined during processing of metal samples in a plasma reactor. The reactor was a Pyrex tube with an inner diameter of 11 cm and a length of 30 cm. Plasma was created by an inductively coupled radiofrequency generator operating at a frequency of 27.12 MHz and output power up to 500 W. The O density was measured at the edge of the glass tube with a copper fiber optics catalytic probe. The O atom density in the empty tube depended on pressure and was between 4 and 7 x 10{sup 21} m{sup -3}. The maximum O density was at a pressure of about 150 Pa, while the dissociation fraction of O{sub 2} molecules was maximal at the lowest pressure and decreased with increasing pressure. At about 300 Pa it dropped below 10%. The measurements were repeated in the chamber loaded with different metallic samples. In these cases, the density of oxygen atoms was lower than that in the empty chamber. The results were explained by a drain of O atoms caused by heterogeneous recombination on the samples.

  6. Determination of mycophenolic acid in human plasma by ultra performance liquid chromatography tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Vivek Upadhyay

    2014-06-01

    Full Text Available A simple, sensitive and high throughput ultra performance liquid chromatography tandem mass spectrometry method has been developed for the determination of mycophenolic acid in human plasma. The method involved simple protein precipitation of MPA along with its deuterated analog as an internal standard (IS from 50 µL of human plasma. The chromatographic analysis was done on Acquity UPLC C18 (100 mm×2.1 mm, 1.7 µm column under isocratic conditions using acetonitrile and 10 mM ammonium formate, pH 3.00 (75:25, v/v as the mobile phase. A triple quadrupole mass spectrometer operating in the positive ionization mode was used for quantitation. In-source conversion of mycophenolic glucuronide metabolite to the parent drug was selectively controlled by suitable optimization of cone voltage, cone gas flow and desolvation temperature. The method was validated over a wide concentration range of 15–15000 ng/mL. The mean extraction recovery for the analyte and IS was >95%. Matrix effect expressed as matrix factors ranged from 0.97 to 1.02. The method was successfully applied to support a bioequivalence study of 500 mg mycophenolate mofetil tablet in 72 healthy subjects.

  7. Determination by high-performance liquid chromatography of phenylbutazone in samples of plasma from fighting bulls.

    Science.gov (United States)

    Marti, M I González; Sánchez, González C I; Jiménez, Hernández; Cachán, García; Castro, de Cabo M J; Cuadrado, A L Garzón

    2002-03-25

    The purpose of this study was to investigate the possible presence of phenylbutazone in plasma samples from fighting bulls killed in 2nd and 3rd category bullrings in the province of Salamanca (Spain) in 1998, 1999 and 2000. For quantitative and qualitative determination, a high-performance liquid chromatograph was used, equipped with a photodiode-array detector and setting wavelengths at 240, 254 and 284 nm. The mobile phase optimized for the simultaneous detection of dexamethasone, betamethasone, flunixin and phenylbutazone, was 0.01 M acetic acid pH 3 in methanol (35:65 v/v) at a flow rate of 1 ml/min. Plasma samples were deproteinized with 400 microl of acetonitrile and 20 microl of the supernatant were injected directly into the chromatographic system equipped with a Lichrospher 60 RP select B column and guard column. For the quantitative analysis, standard calibration curves were made in a concentration range between 0.25 and 30 microg/ml, using betamethasone as internal standard. The retention time of phenylbutazone was 8.7 +/- 0.2 min and recovery was 83%. The detection and quantification limits were 0.016 and 0.029, respectively for A=240 nm. The study results show that 17 of the 74 samples analyzed in 1998, 18 of those from 1999 and 10 of those from 2000 were positive for phenylbutazone.

  8. Direct injection HPLC method for the determination of selected benzodiazepines in plasma using a Hisep column.

    Science.gov (United States)

    Pistos, C; Stewart, James T

    2003-12-04

    A direct plasma injection HPLC method has been developed for the determination of selected benzodiazepines (nitrazepam, clobazam, oxazepam, lorazepam). The method uses an analytical hydrophobic shielded phase (Hisep) column equipped with a Hisep guard column, are easy to perform and requires 20 ul of a filtered plasma sample. The chromatographic run time is less than 15 min using a mobile phase of 15:85 v/v acetonitrile-0.18 M ammonium acetate pH 2.5. The method is good for 175 injections before replacement of the guard column. The method was linear in the range 0.5-18 ug ml(-1) (r>0.99, n=6) for the analytes with R.S.D. less than 10.82%. Interday and intraday variability were found to be less than 14%. The limits of detection and quantitation were 0.16 (s/n>3) and 0.5 ug ml(-1) (s/n>10), respectively, for each of the four benzodiazepines.

  9. Is it the egg or the endometrium? Elevated progesterone on day of trigger is not associated with embryo ploidy nor decreased success rates in subsequent embryo transfer cycles

    National Research Council Canada - National Science Library

    Kofinas, Jason D; Mehr, Holly; Ganguly, Nandita; Biley, Yelena; Bochkovsky, Svetlana; McCulloh, David; Grifo, Jamie

    2016-01-01

    The purpose of our study was to determine if progesterone (P4) values on day of trigger affect certain cycle outcome parameters, ploidy status of embryos, as well as pregnancy outcomes in the subsequent first frozen embryo transfer...

  10. Spectrophotometric determination of total proteins in blood plasma: a comparative study among dye-binding methods

    Directory of Open Access Journals (Sweden)

    Dimas Augusto Morozin Zaia

    2005-05-01

    Full Text Available A comparative study between the biuret method (standard method for total proteins and spectrophotometric methods using dyes (Bradford, 3',3",5',5"-tetrabromophenolphthalein ethyl ester-TBPEE, and erythrosin-B was carried out for the determination of total proteins in blood plasma from rats. Bradford method showed the highest sensitivity for proteins and biuret method showed the lowest. For all the methods, the absorbance for different proteins (BSA, casein, and egg albumin was measured and Bradford method showed the lowest variation of absorbance. The concentration of total protein obtained by using Bradford method was not statistically different (p>0.05 from concentration of total protein obtained by the biuret method. But in regard to erythrosin-B and TBPEE methods the concentrations of total protein were statistically different (pA determinação de proteínas totais em plasma sangüíneo é importante em diversas áreas de pesquisa. Um estudo comparativo entre o método de biureto (método padrão para proteínas totais e diversos métodos que utilizam corantes (Bradford, tetrabromofenolftaleína etil éster-TBPEE, e eritrosina-B foi realizado para a determinação de proteínas totais em plasma sangüíneo de ratos. O método de Bradford mostrou a maior sensibilidade para proteínas e o de biureto a menor. Para todos os métodos, as absorbâncias para diferentes proteínas (BSA, caseína, e ovoalbumina foram medidas e o método de Bradford mostrou a menor variação da absorbância. Utilizando o método de Bradford a concentração de proteínas totais obtida não foi estatisticamente diferente (p>0.05 daquela obtida pelo método do biureto. Porém, para os métodos da eritrosina-B e TBPEE as concentrações de proteínas totais foram estatisticamente diferentes (p<0.05 da obtida pelo método de biureto. Portanto o método de Bradford pode ser utilizado no lugar do método de biureto para a determinação de proteínas totais em plasma sangüíneo.

  11. Progesterone binding nano-carriers based on hydrophobically modified hyperbranched polyglycerols

    Science.gov (United States)

    Alizadeh Noghani, M.; Brooks, D. E.

    2016-02-01

    results provide evidence to justify more detailed studies of interactions with biological systems, both single cells and in animal models. Electronic supplementary information (ESI) available: Fig. S-1: chemical structure of progesterone (Pro). Fig. S-2: 1H NMR spectrum of HPG-C8-MPEG. Fig. S-3: GPC chromatogram of HPG-C8-MPEG. Fig. S-4: 1H NMR spectrum of HPG-C12-MPEG. Fig. S-5: GPC chromatogram of HPG-C8-MPEG. Fig. S-6: FTIR spectrum of HPG-C8-MPEG. Fig. S-7: inverse-gated 13C NMR spectrum of HPG-C8-MPEG in methanol-d4. Fig. S-8: semi-log plot to determine initial rapid release kinetics for HPG-C8-MPEG/Pro in PBS. Fig. S-9: semi-log plot to determine secondary slow release kinetics for HPG-C8-MPEG/Pro in PBS. Fig. S-10: semi-log plot illustrating the kinetics of Pro release from HPG-C8-MPEG/Pro in plasma. Fig. S-11: dependence of k1 and Vp - Va. Fig. S-12: correlation between the maximum binding capacity of HPG-Cn-MPEG polymeric systems for binding Pro and their total mass of alkyl carbon external to the oxygen (R2 = 0.77 and p right). The minor population of larger particles was reduced in diameter by Pro binding, illustrated above, consistent with an earlier report.11 See DOI: 10.1039/c5nr08175k

  12. Progesterone attenuates depressive behavior of younger and older adult C57/BL6, wildtype, and progesterone receptor knockout mice

    OpenAIRE

    Frye, Cheryl A.

    2011-01-01

    Progesterone may have actions independent of intracellular progestin receptors (PRs) to influence depressive behavior. To investigate this, we examined effects of progesterone (P; 10 mg/kg, SC) on the depressive behavior of mice in the forced swim test (FST). In Experiment 1, subjects were 4 to 6 months old, intact or ovariectomized (OVX) female and intact or gonadectomized (GDX) male, C57/BL6 mice. Progesterone reduced depressive behavior of young diestrous and OVX mice but male mice were im...

  13. Expression of estrogen and progesterone receptors in papillary thyroid carcinoma

    Science.gov (United States)

    Jalali-Nadoushan, Mohammad-Reza; Amirtouri, Reza; Davati, Ali; Askari, Samaneh; Siadati, Sepideh

    2016-01-01

    Background: Papillary thyroid carcinoma (PTC), occurs mostly in women and sex hormones may play a role in the pathogenesis and clinical course. The objective of this study was to determine the status and prevalence of estrogen and progesterone receptors in PTC with regard to age, gender, tumor size and lymph node involvement. Methods: Immunohistochemical stains were performed on 92 tissue blocks of PTC for estrogen receptor (ER) and progesterone receptor (PR) expression in tumor cells. Chi-square test and Mann-Whitney U test were used to determine statistical difference using statistical software SPSS. Results: The mean age of patients was 39.32±1.7 years (range 13-80) with 79(85.9%) women and 13 (14.1%) men. Lymph node involvement was seen in 76.1% of patients. The average tumor size was 3.6±2.21 cm. The rate of ER and PR expression were 46.75% and 5.6%, respectively. ER expression for females was higher than males (P=0.014), but no relation was found between males and females in PR expression (P=0.7). Also there was no statistical difference between ER and PR expression with respect to age, lymph node involvement and tumor size. Conclusion: Our study showed higher ER expression in females than males with PTC. No relation was found between the expression of these receptors and age of presentation, lymph node involvement and tumor size. Further investigation is required to determine the prognostic importance of ER and PR in PTC.

  14. Changes in rat uterine and cervical phospholipase A2 activity following progesterone agonist or antagonist administration at term.

    Science.gov (United States)

    Kurusu, Shiro; Ishii, Shizuka; Kawaminami, Mitsumori

    2007-04-01

    Our previous study revealed that a fall in plasma progesterone (P(4)) level was associated with a transient increase in cytosolic phospholipase A(2) (PLA(2)) activity and prostaglandin F(2)alpha level in the rat uterus and cervix during natural parturition. This study determined the changes in the PLA(2) activities during modulated occurrence of delivery by P(4) antagonist or agonist late in pregnancy. In rats undergoing P(4) antagonist-induced preterm delivery, the PLA(2) activities of both uterine and cervical cytosol significantly decreased 12 h after the challenge and tended to be attenuated within 72 h. The plasma P(4) level altered in a similar pattern. Blockade of delivery by chronic treatment with P(4) agonist was not associated with changes in uterine PLA(2) activity compared with that in normally delivering rats, although there was a persistent rise in cervical PLA(2) activity. The obtained data indicates that the PLA(2) activities in rat uterine and cervical cytosol are not regulated solely by P(4) and that delivery can occur without activation of this enzyme.

  15. Seasonal patterns of circulating progesterone and prolactin and response to bromocriptine in the female tammar Macropus eugenii.

    Science.gov (United States)

    Tyndale-Biscoe, C H; Hinds, L A

    1984-01-01

    Concentrations of prolactin and progesterone in the plasma of female tammars (Macropus eugenii) were measured during lactational quiescence and seasonal quiescence and during the period of natural resumption of reproduction after the summer solstice in December. Prolactin concentrations were consistently low (less than 40 ng/ml) during the period of declining day length and consistently elevated (greater than 40 ng/ml) during the period of increasing day length. Basal levels of progesterone were lowest (118.9 +/- 9.1 pg/ml) at the winter solstice and highest (244.1 +/- 21.0 pg/ml) at the summer solstice. Treatment with bromocriptine (CB154) did not depress prolactin levels at either time of the year but during February to June a single injection of 5 mg/kg body wt induced development of the quiescent corpus luteum (CL) and the diapausing embryo was reactivated. In February and March the level of progesterone increased in association with the developing CL, but after treatment at the winter solstice in June plasma progesterone did not increase although pregnancy was successfully completed. From September through November only 1 of 50 females responded to bromocriptine. None responded to bromocriptine given at the summer solstice in December, but all these animals spontaneously reactivated 2 weeks later. The difference in response of female tammars to bromocriptine treatment in the two halves of the year suggests that different endocrine controls operate in lactational and seasonal quiescence and that the latter is more complex.

  16. Rapid Determination of Isomeric Benzoylpaeoniflorin and Benzoylalbiflorin in Rat Plasma by LC-MS/MS Method

    Directory of Open Access Journals (Sweden)

    Chuanqi Zhou

    2017-01-01

    Full Text Available Benzoylpaeoniflorin (BP is a potential therapeutic agent against oxidative stress related Alzheimer’s disease. In this study, a more rapid, selective, and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS method was developed to determine BP in rat plasma distinguishing with a monoterpene isomer, benzoylalbiflorin (BA. The method showed a linear response from 1 to 1000 ng/mL (r>0.9950. The precision of the interday and intraday ranged from 2.03 to 12.48% and the accuracy values ranged from −8.00 to 10.33%. Each running of the method could be finished in 4 minutes. The LC-MS/MS method was validated for specificity, linearity, precision, accuracy, recovery, and stability and was found to be acceptable for bioanalytical application. Finally, this fully validated method was successfully applied to a pharmacokinetic study in rats following oral administration.

  17. High-performance liquid chromatographic determination of vertilmicin in rat plasma using sensitive fluorometric derivatization.

    Science.gov (United States)

    Liu, Zhen; Sha, Yunfei; Huang, Taomin; Yang, Bei; Duan, Geng-Li

    2005-12-15

    A sensitive and reliable high-performance liquid chromatographic method was developed for the determination of vertilmicin in rat plasma. Derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by C(18) reversed-phase chromatography allowed the fluorimetric detection of vertilmicin. Optimal conditions for the derivatization of vertilmicin are described. The limit of quantification was 0.02 mg/L. The pharmacokinetics of vertilmicin was studied in 24 rats following intramuscular injection (i.m.) of different doses (4, 8, 16, 32 mg/kg of body weight). The pharmacokinetic parameter values were estimated by use of 3P97 program. In this study, we assessed the dose proportionality of vertilmicin after single intramuscular injection doses and obtained new information on the pharmacokinetics of the compound.

  18. DETERMINATION OF FLUCONAZOLE IN HUMAN PLASMA BY REVERSE PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY.

    Science.gov (United States)

    Safaei, Zahra; Alipour, Eskandar; Shafaati, Alireza; Zarghp, Afshin

    2015-01-01

    A rapid, simple and reproducible high performance liquid chromatographic method was developed and validated for determination of fluconazole in human plasma. The separation was performed on MZ C8 column (125 x 4 mm, 5 µm) using acetonitrile - potassium dihydrogen phosphate buffer (15 : 85, v/v), pH 3.0, as the mobile phase at a flow rate of 1.5 mL/min. The wavelength was set at 261 nm. The assay enables the measurement of fluconazole for therapeutic drug monitoring with a minimum quantification limit of 20 ng/mL. The method involves simple, protein precipitation procedure and analytical recovery was complete. The calibration curve was linear over the concentration range 0.1-4 µg/mL. The coefficients of variation for inter-day and intra-day assay were found to be less than 10%.

  19. Improved Cd determination in glasses by laser ablation inductively coupled plasma mass spectrometry using nitrogen as a matrix modifier

    Institute of Scientific and Technical Information of China (English)

    Qian Ni; Zhao Chu Hu; Zheng Yu Bao; Ya Feng Zhang

    2009-01-01

    The addition of 5-10 mL min-1 nitrogen to the central channel of plasma in Laser ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) increases the sensitivities of Cd by a factor of 3 and decreases oxide interferences by one order of magnitude, which allows the direct analysis of trace levels of Cd in glass samples. This simple method shows a great potential for the direct determination of Cd in various kinds of samples.

  20. Electron density in amplitude modulated microwave atmospheric plasma jet as determined from microwave interferometry and emission spectroscopy

    Science.gov (United States)

    Faltýnek, J.; Hnilica, J.; Kudrle, V.

    2017-01-01

    Time resolved electron density in an atmospheric pressure amplitude modulated microwave plasma jet is determined using the microwave interferometry method, refined by numerical modelling of the propagation of non-planar electromagnetic waves in the vicinity of a small diameter, dense collisional plasma filament. The results are compared to those from the Stark broadening of the {{\\text{H}}β} emission line. Both techniques show, both qualitatively and quantitatively, a similar temporal evolution of electron density during one modulation period.

  1. Progesterone suppresses triple-negative breast cancer growth and metastasis to the brain via membrane progesterone receptor α.

    Science.gov (United States)

    Zhou, Li; Zhou, Wei; Zhang, Hongwei; Hu, Yan; Yu, Lei; Zhang, Yufei; Zhang, Yanli; Wang, Shuang; Wang, Peng; Xia, Wei

    2017-09-01

    Progesterone plays an important role in mammary epithelial cell proliferation and differentiation. Evidence from experimental and clinical studies indicates that progesterone is a risk factor for breast cancer under certain conditions through binding nuclear progesterone receptor (PR). These mechanisms, however, are not applicable to triple-negative breast cancer (TNBC) due to the lack of PR in these cancers. In this study, we demonstrate that membrane progesterone receptor α (mPRα) is expressed in TNBC tissues and the expression level of mPRα is negatively associated with the TNM stage. We found that progesterone suppressed the growth, migration and invasion of mPRα+ human TNBC cells in vitro, which was neither mediated by PR nor by PR membrane component 1 (PGRMCl). Notably, these effects exerted by progesterone were significantly blocked by shRNA specific to mPRα. Moreover, the knockdown of mPRα expression impaired the inhibitory effects of progesterone on mPRα+ tumor growth and metastasis in vivo. These data collectively indicate that progesterone suppresses TNCB growth and metastasis via mPRα, which provides evidence of the anti-neoplastic effects of progesterone-mPRα pathway in the treatment of human TNBC.

  2. Mifepristone acts as progesterone antagonist of non-genomic responses but inhibits phytohemagglutinin-induced proliferation in human T cells.

    Science.gov (United States)

    Chien, C H; Lai, J N; Liao, C F; Wang, O Y; Lu, L M; Huang, M I; Lee, W F; Shie, M C; Chien, E J

    2009-08-01

    Progesterone is an endogenous immunomodulator that suppresses T cell activation during pregnancy. The stimulation of membrane progesterone receptors (mPRs) would seem to be the cause of rapid non-genomic responses in human peripheral T cells, such as an elevation of intracellular calcium ([Ca(2+)](i)) and decreased intracellular pH (pH(i)). Mifepristone (RU486) produces mixed agonist/antagonist effects on immune cells compared with progesterone. We explored whether RU486 is an antagonist to mPRs and can block rapid non-genomic responses and the induction by phytohemagglutinin (PHA) of cell proliferation. Human male peripheral T cell responses in terms of pH(i) and [Ca(2+)](i) changes were measured using the fluorescent dyes, 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) and fura-2, respectively. Expression of mPR mRNA was determined by RT-PCR analysis. Cell proliferation and cell toxicity were determined by [(3)H]-thymidine incorporation and MTT assay, respectively. The mRNAs of mPRalpha, mPRbeta and mPRgamma were expressed in T cells. RU486 blocked progesterone-mediated rapid responses including, the [Ca(2+)](i) increase and pH(i) decrease, in a dose related manner. RU486 did not block, but enhanced, the inhibitory effect of progesterone on PHA induced cell proliferation. RU486 alone inhibited proliferation induced by PHA and at >25 microM seems to be cytotoxic against resting T cells (P < 0.01). RU486 is antagonistic to the rapid mPR-mediated non-genomic responses, but is synergistic with progesterone with respect to the inhibition of PHA-induced cell proliferation. Our findings shine new light on RU486's clinical application and how this relates to the non-genomic rapid physiological responses caused by progesterone.

  3. Dielectric relaxation and crystallization of ultraviscous melt and glassy states of aspirin, ibuprofen, progesterone, and quinidine.

    Science.gov (United States)

    Johari, G P; Kim, S; Shanker, Ravi M

    2007-05-01

    Molecular relaxation in ultraviscous melt and glassy states of aspirin, ibuprofen, progesterone, and quinidine has been studied by dielectric spectroscopy. The asymmetric relaxation spectra is characterized by the Kohlrausch distribution parameter of 0.46 +/- 0.02 for aspirin to 0.67 +/- 0.02 for progesterone. The dielectric relaxation time varies with the temperature, T, according to the Vogel-Fulcher-Tammann Equation, log(10)(tau(0)) = A(VFT) + [B(VFT)/(T - T(0))], where A(VFT), B(VFT), and T(0) are empirical constants. The extrapolated tau(0) at calorimetric glass-softening temperature is close to the value expected. The equilibrium permittivity, epsilon(0), is lowest for ibuprofen which indicates an antiparallel orientation of dipoles in its liquid's hydrogen-bonded structure. A decrease in epsilon(0) with time shows that ultraviscous aspirin, progesterone, and quinidine begin to cold-crystallize at a relatively lower temperature than ibuprofen. epsilon(0) of the cold-crystallized phases are, 4.7 for aspirin at 290 K, 2.55 for ibuprofen at 287 K, 2.6 for progesterone at 320 K, and 3.2 for quinidine at 375 K. It is argued that hydrogen-bonding, the Kohlrausch parameter, extent of localized motions and the long-range diffusion times all determine the physical and chemical stability of an amorphous pharmaceutical during storage. (c) 2007 Wiley-Liss, Inc. and the American Pharmacists Association.

  4. Structural and dynamical aspects of the distant magnetotail determined from ISEE-3 plasma measurements

    Energy Technology Data Exchange (ETDEWEB)

    Hones, E.W.; Zwicki, R.D.; Fritz, T.A.; Bame, S.J.

    1986-10-01

    Large lateral displacements of the magnetotail at ISEE-3 orbital distances can result from the angular variations of solar wind flow that occur frequently. To determine the tail's shape and structure from the ISEE-3 measurements it is desirable to determine quantitatively the tail's displacement during such measurements. Three-dimensional solar wind measurements by IMP-8 have been used together with a new coordinate system, the Geocentric Solar Wind (GSW) system, which reflects solar wind direction changes (as well as earth dipole motion), to estimate the location of ISEE-3 within the tail cross-section. Preliminary tests suggest that at least those changes of satellite location within the tail resulting from long term (few hour) solar wind changes can be determined by the method. Energetic ion spectra, measured in plasmoids by ISEE-3 in the distant tail and by ISEE-2 at the time of the plasmoid's departure from the near tail, have the same exponential form, distinctly different from the power law form that has earlier been ascribed to plasma sheet ion spectra. The ions are trapped in the closed loops of the plasmoid and the major anisotropy that they show is due to their convective motion with the moving plasmoid, not to field-aligned streaming.

  5. The validation of the Z-Scan technique for the determination of plasma glucose

    Science.gov (United States)

    Alves, Sarah I.; Silva, Elaine A. O.; Costa, Simone S.; Sonego, Denise R. N.; Hallack, Maira L.; Coppini, Ornela L.; Rowies, Fernanda; Azzalis, Ligia A.; Junqueira, Virginia B. C.; Pereira, Edimar C.; Rocha, Katya C.; Fonseca, Fernando L. A.

    2013-11-01

    Glucose is the main energy source for the human body. The concentration of blood glucose is regulated by several hormones including both antagonists: insulin and glucagon. The quantification of glucose in the blood is used for diagnosing metabolic disorders of carbohydrates, such as diabetes, idiopathic hypoglycemia and pancreatic diseases. Currently, the methodology used for this determination is the enzymatic colorimetric with spectrophotometric. This study aimed to validate the use of measurements of nonlinear optical properties of plasma glucose via the Z-Scan technique. For this we used samples of calibrator patterns that simulate commercial samples of patients (ELITech ©). Besides calibrators, serum glucose levels within acceptable reference values (normal control serum - Brazilian Society of Clinical Pathology and Laboratory Medicine) and also overestimated (pathological control serum - Brazilian Society of Clinical Pathology and Laboratory Medicine) were used in the methodology proposal. Calibrator dilutions were performed and determined by the Z-Scan technique for the preparation of calibration curve. In conclusion, Z-Scan method can be used to determinate glucose levels in biological samples with enzymatic colorimetric reaction and also to apply the same quality control parameters used in biochemistry clinical.

  6. Determination of myo-inositol hexakisphosphate (phytate) in urine by inductively coupled plasma atomic emission spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Grases, F.; Perello, J.; Isern, B.; Prieto, R.M

    2004-05-10

    Myo-inositol hexakisphosphate (phytate) is a substance present in urine with an important role in preventing calcium renal calculi development. In spite of this, the use of urinary phytate levels on stone-formers' evaluation and treatment is still notably restricted as a consequence of the enormous difficulty to analyze this substance in urine. In this paper, a simple procedure for routinary urinary phytate determination based on phosphorus determination through inductively coupled plasma atomic emission spectrometry is described. The method only requires a previous separation of phytate from other components by column anion exchange chromatography. The working linear range used was 0-2 mg l{sup -1} phosphorus (0-7 mg l{sup -1} phytate). The limit of detection was 64 {mu}g l{sup -1} of phytate and the limit of quantification was 213 {mu}g l{sup -1}. The relative standard deviation (R.S.D.) for 1.35 mg l{sup -1} phytate was 2.4%. Different urine samples were analyzed using an alternative analytical methodology based on gas chromatography (GC)/mass detection used for inositol determination (phytate was previously hydrolyzed), resulting both methods comparable using as criterion to assess statistical significance P<0.05.

  7. HPTLC Method for the Determination of Paracetamol, Pseudoephedrine and Loratidine in Tablets and Human Plasma.

    Science.gov (United States)

    Farid, Nehal Fayek; Abdelaleem, Eglal A

    2016-04-01

    A sensitive, accurate and selective high performance thin layer chromatography (HPTLC) method was developed and validated for the simultaneous determination of paracetamol (PAR), its toxic impurity 4-aminophenol (4-AP), pseudoephedrine HCl (PSH) and loratidine (LOR). The proposed chromatographic method has been developed using HPTLC aluminum plates precoated with silica gel 60 F254 using acetone-hexane-ammonia (4:5:0.1, by volume) as a developing system followed by densitometric measurement at 254 nm for PAR, 4-AP and LOR, while PSH was scanned at 208 nm. System suitability testing parameters were calculated to ascertain the quality performance of the developed chromatographic method. The method was validated with respect to USP guidelines regarding accuracy, precision and specificity. The method was successfully applied for the determination of PAR, PSH and LOR in ATSHI(®) tablets. The three drugs were also determined in plasma by applying the proposed method in the ranges of 0.5-6 µg/band, 1.6-12 µg/band and 0.4-2 µg/band for PAR, PSH and LOR, respectively. The results obtained by the proposed method were compared with those obtained by a reported HPLC method, and there was no significance difference between both methods regarding accuracy and precision.

  8. Application of new membrane selective electrodes for the determination of drotaverine hydrochloride in tablets and plasma.

    Science.gov (United States)

    El-Saharty, Y S; Metwaly, F H; Refaat, M; El-Khateeb, S Z

    2006-06-07

    The construction and electrochemical response characteristics of poly vinyl chloride (PVC) membrane sensors for the determination of drotaverine hydrochloride were described. The sensors are based on the use of the ion association complexes of drotaverine cation with sodium phosphotungestate (Dro-PTA) or ammonium reineckate (Dro-R) counter anions as ion exchange sites in the PVC matrix. The performance characteristics of these sensors, which were evaluated according to IUPAC recommendations, reveal a fast, stable and linear response for drotaverine over the concentration range 10(-5) to 10(-2) M with cationic slopes of 49.55 and 51.36 mV per concentration decade. The direct potentiometric determination of drotaverine hydrochloride using the proposed sensors gave average recoveries of 99.95+/-0.71 and 100.04+/-0.60 for Dro-PTA and Dro-R, respectively. The sensors are used for determination of drotaverine hydrochloride in tablets, in its mixture with caffeine and paracetamol and in plasma. Validation of the method shows suitability of the proposed sensors for use in the quality control assessment of drotaverine hydrochloride. The developed method was found to be simple, accurate and precise when compared with a reported HPLC method.

  9. Chemiluminescence Determination of Local Anaesthetic Mepivacaine in Human Plasma and Pharmaceuticals.

    Science.gov (United States)

    Mokhtari, Ali

    2016-12-01

    In this study, a new method has been developed for the simple determination of mepivacaine (carbocaine). The method is based on the enhancement effect of mepivacaine in the chemiluminescence (CL) reaction of tris(1,10 phenanthroline) ruthenium(II) with cerium(IV). A mechanism for the CL reaction has been proposed on the basis of UV-Vis, fluorescent and CL spectra. Under optimum conditions, the CL intensity was proportional to the concentration of the drug in solution over the range 0.45-226.25 μg mL-1 (R2 = 0.9996). The LOD was 0.34 μg mL-1 (S/N = 3). LOD was about 10 times lower than the therapeutic concentration of mepivacaine. The percent of relative standard deviation for determination of 11 replicates at level of 9.05 μg mL-1 and 90.50 μg mL-1 of mepivacaine were 1.8 and 3.7%, respectively. The proposed method was applied successfully for the determination of mepivacaine in human plasma and injectable solutions.

  10. Uterine involution and progesterone level during the postpartum period in Barbary ewes in north Libya

    Directory of Open Access Journals (Sweden)

    M.S. Medan

    2015-02-01

    Full Text Available The objectives of the present study were to determine the time of uterine involution and ovarian activity using ultrasound examination and progesterone assay. Weekly progesterone levels were measured starting one week postpartum until two weeks after the 1st postpartum estrus in Barbary ewes lambed during winter in AL-Bayda city, north of Libya. A total of 15 Barbary ewes were used in the present study distributed in three groups according to the month of lambing as group 1 (lambed in January, group 2 (lambed in February and group 3 (lambed in March. Ewes were examined weekly by trans-rectal ultrasound to check involution of the uterus starting one week after lambing until complete uterine involution. Blood samples were collected from the jugular vein, and serum was separated and stored at ˗20 ºC until measuring progesterone using ELISA. Results showed that uterine involution completed at day 35 postpartum in groups 1 and 2, while it occurred at day 28 in group 3. The mean progesterone level was basal (less than 1 ng/ml for a long period and started to increase at days 119, 99 and 77 postpartum in group 1, 2 and 3, respectively. One ewe did not show estrus at all during the period of study in group 2 and there were no growing follicles on their ovaries. The obtained results indicate that, uterine involution as determined by ultrasound completed earlier in ewes lambed in March than those lambed in February or January. Also, progesterone level and ultrasound examination showed that there was no ovarian activity for a longtime after parturition indicating that reproduction in Barbary ewes tends to be seasonal in AL-Bayda city, north Libya.

  11. Uterine involution and progesterone level during the postpartum period in Barbary ewes in north Libya.

    Science.gov (United States)

    Medan, M S; El-Daek, T

    2015-01-01

    The objectives of the present study were to determine the time of uterine involution and ovarian activity using ultrasound examination and progesterone assay. Weekly progesterone levels were measured starting one week postpartum until two weeks after the 1(st) postpartum estrus in Barbary ewes lambed during winter in AL-Bayda city, north of Libya. A total of 15 Barbary ewes were used in the present study distributed in three groups according to the month of lambing as group 1 (lambed in January), group 2 (lambed in February) and group 3 (lambed in March). Ewes were examined weekly by trans-rectal ultrasound to check involution of the uterus starting one week after lambing until complete uterine involution. Blood samples were collected from the jugular vein, and serum was separated and stored at -20 °C until measuring progesterone using ELISA. Results showed that uterine involution completed at day 35 postpartum in groups 1 and 2, while it occurred at day 28 in group 3. The mean progesterone level was basal (less than 1 ng/ml) for a long period and started to increase at days 119, 99 and 77 postpartum in group 1, 2 and 3, respectively. One ewe did not show estrus at all during the period of study in group 2 and there were no growing follicles on their ovaries. The obtained results indicate that, uterine involution as determined by ultrasound completed earlier in ewes lambed in March than those lambed in February or January. Also, progesterone level and ultrasound examination showed that there was no ovarian activity for a longtime after parturition indicating that reproduction in Barbary ewes tends to be seasonal in AL-Bayda city, north Libya.

  12. A Randomized Trial of Progesterone in Women with Recurrent Miscarriages

    NARCIS (Netherlands)

    Coomarasamy, Arri; Williams, Helen; Truchanowicz, Ewa; Seed, Paul T; Small, Rachel; Quenby, Siobhan; Gupta, Pratima; Dawood, Feroza; Koot, Yvonne E M; Bender Atik, Ruth; Bloemenkamp, Kitty W M; Brady, Rebecca; Briley, Annette L; Cavallaro, Rebecca; Cheong, Ying C; Chu, Justin J; Eapen, Abey; Ewies, Ayman; Hoek, Annemieke; Kaaijk, Eugenie M; Koks, Carolien A M; Li, Tin-Chiu; MacLean, Marjory; Mol, Ben W; Moore, Judith; Ross, Jackie A; Sharpe, Lisa; Stewart, Jane; Vaithilingam, Nirmala; Farquharson, Roy G; Kilby, Mark D; Khalaf, Yacoub; Goddijn, Mariette; Regan, Lesley; Rai, Rajendra

    2015-01-01

    BACKGROUND: Progesterone is essential for the maintenance of pregnancy. However, whether progesterone supplementation in the first trimester of pregnancy would increase the rate of live births among women with a history of unexplained recurrent miscarriages is uncertain. METHODS: We conducted a mult

  13. A Randomized Trial of Progesterone in Women with Recurrent Miscarriages

    NARCIS (Netherlands)

    Coomarasamy, A.; Williams, H.; Truchanowicz, E.; Seed, P. T.; Small, R.; Quenby, S.; Gupta, P.; Dawood, F.; Koot, Y. E. M.; Atik, R. Bender; Bloemenkamp, K. W. M.; Brady, R.; Briley, A. L.; Cavallaro, R.; Cheong, Y. C.; Chu, J. J.; Eapen, A.; Ewies, A.; Hoek, A.; Kaaijk, E. M.; Koks, C. A. M.; Li, T. -C.; MacLean, M.; Mol, B. W.; Moore, J.; Ross, J. A.; Sharpe, L.; Stewart, J.; Vaithilingam, N.; Farquharson, R. G.; Kilby, M. D.; Khalaf, Y.; Goddijn, M.; Regan, L.; Rai, R.

    2015-01-01

    BACKGROUND Progesterone is essential for the maintenance of pregnancy. However, whether progesterone supplementation in the first trimester of pregnancy would increase the rate of live births among women with a history of unexplained recurrent miscarriages is uncertain. METHODS We conducted a multic

  14. Determination of lamivudine in human plasma by HPLC and its use in bioequivalence studies.

    Science.gov (United States)

    Kano, Eunice Kazue; dos Reis Serra, Cristina Helena; Koono, Eunice Emiko Mori; Andrade, Simone Schramm; Porta, Valentina

    2005-06-13

    A simple, accurate, precise and sensitive high-performance liquid chromatographic (HPLC) method with ultraviolet detection was developed to quantificate lamivudine (3-TC) in human plasma samples from bioequivalence studies. 3-TC and stavudine (internal standard, I.S.) were extracted from 0.5 ml of human plasma by acetonitrile protein precipitation. The method was validated over a concentration range of 0.05-3.00 microg/ml and used in a bioequivalence trial between two lamivudine formulations, to assess its usefulness in this kind of study. FURP-lamivudine (Fundação para o Remédio Popular, Brazil, as test formulation) and Epivir (GlaxoSmithKline, Brazil, as reference formulation) were evaluated following a single 150 mg oral dose to 24 healthy volunteers of both genders. The dose was administered after an overnight fast according to a two-way crossover design. Bioequivalence between the products was determined by calculating 90% confidence intervals (90% CI) for the ratio of Cmax, AUC0-t and AUC0-inf values for the test and reference products, using logarithmic transformed data. The 90% confidence intervals for the ratio of Cmax (0.86-1.06), AUC0-t (0.96-1.04) and AUC0-inf (0.97-1.05) values for the test and reference products are within the 0.80-1.25 interval proposed by FDA and EMEA. It was concluded that the two 3-TC formulations are bioequivalent in their rate and extent of absorption, and thus, may be used interchangeably.

  15. Development and Validation of a HPLC Method for Determination of Pefloxacin in Tablet and Human Plasma

    Directory of Open Access Journals (Sweden)

    Shahnaz Gauhar

    2009-03-01

    Full Text Available Objective(sDeveloping and validating a simple, efficient, reproducible and economic reversed phase high performance liquid chromatographic (RP-HPLC method for the quantitative determination of pefloxacin in bulk material, tablets and in human plasma. Materials and MethodsA shim-pack CLC-ODS column and a mobile phase constituting acetonitrile: 0.025 M phosphoric acid solution (13:87 v/v, pH 2.9 adjusted with KOH were used. The flow rate was 1 ml/min and the analyses performed using ultraviolet (UV detector at a wavelength of 275 nm using acetaminophen as an internal standard.ResultsThe developed method showed good resolution between pefloxacin and acetaminophen. It was selective to pefloxacin and able to resolve the drug peak from internal standard and from formulation excipients. The percentage of coefficient variation (CV of the retention times and peak areas of pefloxacin from the six consecutive injections were 0.566% and 0.989%, respectively. The results showed that the peak area responses are linear within the concentration range of 0.125 µg/ml-12 µg/ml (R2= 0.9987. The limits of detection (LOD and limits of quantitation (LOQ for pefloxacin were 0.03125 µg/ml and 0.125 µg/ml. The intra-day and inter-day variation, RSD were 0.376-0.9056 and 0.739-0.853 respectively; also, inter-day variation with relative standard deviation (RSD were 0.1465-0.821 in plasma. The accuracy results of 70%, 100%, and 130% drugs were 100.72%, 100.34%, and 100.09%, respectively.ConclusionThe method is linear, quantitative, reproducible and could be used as a more convenient, efficient and economical method for the trace analysis of drug in biological fluids, in raw material and tablets.

  16. Separation and Determination of Methylnaltrexone in Human Plasma Samples After Oral Administration by HPLC Coupled with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    WANG An-bao; Joseph F. Foss; YUAN Chun-su; Joachim Osinski

    2005-01-01

    A high performance liquid chromatography(HPLC) method coupled with electrochemical detection and solid phase extraction is described for the separation and determination of methylnaltrexone(MNTX), a quaternary opioid antagonist, in human clinical plasma samples after oral administration. Linearity of the standard curve for MNTX was found in the range of 4.0_150 ng/mL and was statistically conformed. The correlation coefficient(r2) and calibration equation obtained from linear regression analysis are 0.9999 and Y=54.27X-0.22, where Y and X represent the peak area and concentration of MNTX, respectively. The detection limit of MNTX under the present experimental conditions is 2.0 ng/mL by estimating at a ratio of 3 of signal to noise. The mean recovery of MNTX in human plasma is higher than 97%. The analytical method was applied to the pharmacokinetic determination of MNTX after single dose oral administration. These data demonstrate that the change of MNTX plasma concentration versus time is obvious. MNTX level of plasma reaches to a plateau between 45 to 120 minutes and then falls slowly. The content of MNTX in plasma sample maintains at an obviously detectable level after twelve hours of oral administration. The pharmacokinetic parameters for a single dose of 19.2 mg/kg in plasma are cmax=206.42(±16.53) ng/mL and tmax=60 min.

  17. Highly sensitive spectrofluorimetric method for determination of certain aminoglycosides in pharmaceutical formulations and human plasma.

    Science.gov (United States)

    Omar, Mahmoud A; Nagy, Dalia M; Hammad, Mohamed A; Aly, Alshymaa A

    2013-06-01

    A simple, reliable, highly sensitive and selective spectrofluorimetric method has been developed for determination of certain aminoglycosides namely amikacin sulfate, tobramycin, neomycin sulfate, gentamicin sulfate, kanamycin sulfate and streptomycin sulfate. The method is based on the formation of a charge transfer complexes between these drugs and safranin in buffer solution of pH 8. The formed complexes were quantitatively extracted with chloroform under the optimized experimental conditions. These complexes showed an excitation maxima at 519-524 nm and emission maxima at 545-570 nm. The calibration plots were constructed over the range of 4-60 pg mL(-1) for amikacin, 4-50 pg mL(-1) for gentamicin, neomycin and kanamycin, 4-40 pg mL(-1) for streptomycin and 5-50 pg mL(-1) for tobramycin. The proposed method was successfully applied to the analysis of the cited drugs in dosage forms. The proposed method was validated according to ICH and USP guidelines with respect to specificity, linearity, accuracy, precision and robustness. The high sensitivity of the proposed method allowed determination of amikacin and gentamicin in spiked and real human plasma.

  18. Accurate determination of silver nanoparticles in animal tissues by inductively coupled plasma mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Veverková, Lenka [Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, CZ 771 46 Olomouc (Czech Republic); Hradilová, Šárka [Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, CZ 771 46 Olomouc (Czech Republic); Milde, David, E-mail: david.mlde@upol.cz [Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, CZ 771 46 Olomouc (Czech Republic); Panáček, Aleš [Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, CZ 771 46 Olomouc (Czech Republic); Skopalová, Jana [Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, CZ 771 46 Olomouc (Czech Republic); Kvítek, Libor [Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, CZ 771 46 Olomouc (Czech Republic); Petrželová, Kamila [Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, CZ 771 46 Olomouc (Czech Republic); National Reference Laboratory for Chemical Elements, Department of Residues in Kroměříž, State Veterinary Institute Olomouc, Hulínská 2286, CZ 767 60 Kroměříž (Czech Republic); and others

    2014-12-01

    This study examined recoveries of silver determination in animal tissues after wet digestion by inductively coupled plasma mass spectrometry. The composition of the mineralization mixture for microwave assisted digestion was optimized and the best recoveries were obtained for mineralization with HNO{sub 3} and addition of HCl promptly after digestion. The optimization was performed on model samples of chicken meat spiked with silver nanoparticles and a solution of ionic silver. Basic calculations of theoretical distribution of Ag among various silver-containing species were implemented and the results showed that most of the silver is in the form of soluble complexes AgCl{sub 2}{sup −} and AgCl{sub 3}{sup 2−} for the optimized composition of the mineralization mixture. Three animal tissue certified reference materials were then analyzed to verify the trueness and precision of the results. - Highlights: • We performed detailed optimization of microwave assisted digestion procedure of animal tissue used prior to Ag determination by ICP-MS. • We provide basic equilibrium calculations to give theoretical explanation of results from optimization of tested mineralization mixtures. • Results from method validation that was done by analysis of several matrix CRMs are presented.

  19. Determination of fetal rhesus d status by maternal plasma DNA analysis.

    Science.gov (United States)

    Aykut, A; Onay, H; Sagol, S; Gunduz, C; Ozkinay, F; Cogulu, O

    2013-12-01

    In this study, we assessed the feasibility of fetal RhD genotyping by analysis of cell-free fetal DNA(cffDNA) extracted from plasma samples of Rhesus (Rh) D-negative pregnant women by using real-time polymerase chain reaction (PCR). Fetal genotyping was performed on 30 RhD-negative women between 9 and 39 weeks of gestation who were referred to us for invasive testing [amniocentesis/chorionic villi sampling (CVS)]. The fetal RHD genotype was determined based on real-time PCR method. Exons 7 and 10 of the RHD and SRY genes were targeted. Among the pregnant women, 12 were carrying male and 17 were carrying female fetuses. Out of 29 pregnant women, 21 had RhD-positive and nine had RhD-negative fetuses. One sample (case 12, whose blood group was found to be AB Rh [+]) was excluded due to controversial results from repeated serological analyses. All prenatal results were in concordance with postnatal RhD status and fetal sex without false- positive or -negative results. Performing real-time PCR on cffDNA showed accurate, efficient and reliable results, allowing rapid and high throughput non invasive determination of fetal sex and RhD status in clinical samples.

  20. Electrochemistry of raloxifene on glassy carbon electrode and its determination in pharmaceutical formulations and human plasma.

    Science.gov (United States)

    Bagheri, Akbar; Hosseini, Hadi

    2012-12-01

    The electrochemical behavior of raloxifene (RLX) on the surface of a glassy carbon electrode (GCE) has been studied by cyclic voltammetry (CV). The CV studies were performed in various supporting electrolytes, wide range of potential scan rates, and pHs. The results showed an adsorption-controlled and quasi-reversible process for the electrochemical reaction of RLX, and a probable redox mechanism was suggested. Under the optimum conditions, differential pulse voltammetry (DPV) was applied for quantitative determination of the RLX in pharmaceutical formulations. The DPV measurements showed that the anodic peak current of the RLX was linear to its concentration in the range of 0.2-50.0μM with a detection limit of 0.0750μM, relative standard deviation (RSD %) below 3.0%, and a good sensitivity. The proposed method was successfully applied for determination of the RLX in pharmaceutical and human plasma samples with a good selectivity and suitable recovery. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. [Determination of Heavy Metal Elements in Diatomite Filter Aid by Inductively Coupled Plasma Mass Spectrometry].

    Science.gov (United States)

    Nie, Xi-du; Fu, Liang

    2015-11-01

    This study established a method for determining Be, Cr, Ni, As, Cd, Sb, Sn, Tl, Hg and Pb, total 10 heavy metals in diatomite filter aid. The diatomite filter aid was digested by using the mixture acid of HNO₃ + HF+ H₃PO₄ in microwave system, 10 heavy metals elements were determined by inductively coupled plasma mass spectrometry (ICP-MS). The interferences of mass spectrometry caused by the high silicon substrate were optimized, first the equipment parameters and isotopes of test metals were selected to eliminate these interferences, the methane was selected as reactant gas, and the mass spectral interferences were eliminated by dynamic reaction cell (DRC). Li, Sc, Y, In and Bi were selected as the internal standard elements to correct the interferences caused by matrix and the drift of sensitivity. The results show that the detection limits for analyte is in the range of 3.29-15.68 ng · L⁻¹, relative standard deviations (RSD) is less than 4.62%, and the recovery is in the range of 90.71%-107.22%. The current method has some advantages such as, high sensitivity, accurate, and precision, which can be used in diatomite filter aid quality control and safety estimations.

  2. Diffusion effects on the determination of surface catalysis in Inductively Coupled Plasma facility

    Science.gov (United States)

    Viladegut, Alan; Düzel, Ümran; Chazot, Olivier

    2017-03-01

    Atomic recombination is an important process to consider when computing the heat flux transferred to the wall of a re-entry vehicle. Two chemical processes are influencing the species diffusion in the boundary layer surrounding a re-usable Thermal Protection System: gas phase reactions and catalytic recombination at the surface. The coupling between them is not normally taken into account when determining the catalytic recombination coefficient (γ) in plasma facilities. This work aims to provide evidence of such coupling based on both a theoretical analysis and an experimental campaign in the VKI-Plasmatron facility. Recombination coefficient measurements at off-stagnation point configuration on a linear copper calorimeter are provided. An evolution from a high-catalytic to a low-catalytic condition due to the boundary layer growth along the probe is observed. This result is consistent with a parametric analysis carried out using the in-house non-equilibrium boundary layer solver, which shows how the experimentally determined catalysis could be influenced by the amount of gas-phase recombination inside the boundary layer.

  3. Determination of Nickel, Vanadium and Iron in Crude Oil by Three-Phase Plasma Arc Spectrometry

    Science.gov (United States)

    Ghatass, Zekry F.

    2002-12-01

    Three-phase plasma arc (TPPA) with ultrasonic nebulizer is developed for simultaneous determination of trace elements in crude oil samples. Ultrasonic nebulizer is used instead of pneumatic nebulizer in order to minimize the problems caused by the oil viscosity during the operation. This system was used for determination of some trace elements (V, Ni, and Fe) in a crude oil samples. Methyl isobutyl ketone (MIBK) was used to dilute the oil samples. The TPPA instrument offers several advantages including a low cost power supply with no radio frequency, linear dynamic ranges from 4 to 5 of orders of magnitude, and detection limits (0.121, 0.313 and 0.242 (μg/ml) for Ni, V and Fe respectively. The average concentrations were 31 ± 0.45 (μg/ml) for Ni, 40 ± 0.88 (μg/ml) for V and 8 ± 0.74 (μg/ml) for Fe at Balaaiem fields and 2 ± 0.05 (μg/ml) for Ni, 4.8 ± 0.25 (μg/ml) for V and 2 ± 0.10 (μg/ml) for Fe at Wastern Desert fields.

  4. Determination of trace elements in refined gold samples by inductively coupled plasma atomic emission spectrometry

    Directory of Open Access Journals (Sweden)

    Steharnik Mirjana

    2013-01-01

    Full Text Available This paper presents a method for determination the trace contents of silver, copper, iron, palladium, zinc and platinum in refined gold samples. Simultaneous inductively coupled plasma atomic emission spectrometer with radial torch position and cross flow nebulizer was used for determination. In order to compare the different calibration strategies, two sets of calibration standards were prepared. The first set was based on matrix matched calibration standards and the second was prepared without the addition of matrix material. Detection limits for matrix matching calibrations were higher for some elements than those without matrix matching. In addition, the internal standardization method was applied and experiments indicated that indium was the best option as internal standard. The obtained results for gold sample by matrix matching and matrix free calibrations were compared with the obtained results by standard addition method. The accuracy of the methods was tested performing recovery test. Recoveries for spiked sample were in the range of 90-115 %. The accuracy of the methods was also tested by analysis of certified reference material of high pure goldAuGHP1. The best results were achieved by matrix free calibration and standard addition method using indium as internal standard at wavelength of 230 nm. [Projekat Ministarstva nauke Republike Srbije, br. 34024: Development of Technologies for Recycling of Precious, Rare and Associated Metals from Solid Waste in Serbia to High Purity Products

  5. Effects of pregnancy in rats on cysteamine-induced peptic ulcers: role of progesterone.

    Science.gov (United States)

    Montoneri, C; Drago, F

    1997-12-01

    After mating with a sexually active male, groups of female Sprague-Dawley rats were injected with cysteamine (400 mg/kg, subcutaneously) at day 0 (controls), day 5 (early-stage pregnancy), and day 18 (late-stage pregnancy) of pregnancy. In contrast to late-stage pregnancy rats, early-stage pregnancy animals showed a decrease of cysteamine-induced gastroduodenal lesions. When subjected to cysteamine injection, both nonpregnant female and male rats treated for eight days with progesterone (300 microg/rat, subcutaneously) showed a reduced incidence of gastroduodenal lesions. No effect was found in animals pretreated with 17beta-estradiol (200 microg/rat, subcutaneously). Furthermore, increased gastroduodenal mucus levels were found in early-stage pregnancy rats and in animals pretreated with progesterone. These results suggest that increased progesterone plasma levels during early-stage pregnancy may be involved in pregnancy-induced gastric and duodenal protection. This effect may be related to an increase in gastric and duodenal mucus production induced by this hormone.

  6. Activity of phospholipase C and release of prostaglandin F2 alpha by endometrial tissue from ovariectomized ewes receiving progesterone and estradiol.

    Science.gov (United States)

    Raw, R E; Silvia, W J

    1991-03-01

    Progesterone and estradiol interact to regulate secretion of prostaglandin (PG) F2 alpha from the ovine endometrium in response to oxytocin. Two experiments were conducted to determine if these effects were due to changes in activity of phospholipase C or in the second messenger responsive pathways that regulate production of PGF2 alpha. In both experiments, ovariectomized ewes were assigned to one of four treatment groups (control, estradiol, progesterone, progesterone and estradiol). Steroids were administered, in vivo, to mimic the changes that occur during the estrous cycle. On Day 16 of steroid treatment, endometrial tissue was collected and incubated, in vitro, to measure activity of phospholipase C and release of PGF2 alpha. Treatment with progesterone, in vivo, enhanced basal and oxytocin-induced activity of phospholipase C and release of PGF2 alpha, in vitro. Estradiol suppressed oxytocin-induced activity of phospholipase C, both in the presence and absence of progesterone. In contrast to its effects on phospholipase C, estradiol inhibited basal and oxytocin-induced release of PGF2 alpha when administered alone, but not when administered with progesterone. Steroids had similar effects on the release of PGF2 alpha induced by phorbol 12-myristate 13-acetate and A23187. It was concluded that progesterone and estradiol regulate endometrial release of PGF2 alpha by affecting both the activity of phospholipase C and its associated second messenger responsive pathways that may regulate production of PGF2 alpha.

  7. Rapid and Sensitive Method for Quantitative Determination of Lopinavir and Ritonavir in Human Plasma by Liquid Chromatography- Tandem Mass Specrtometry

    Directory of Open Access Journals (Sweden)

    G. A. Temghare

    2009-01-01

    Full Text Available A rapid and sensitive liquid chromatography-mass spectrometric (LC-MS-MS method for the simultaneous determination of lopinavir and ritonavir in human plasma using abacavir as internal standard has been developed and validated. Sample preparation of plasma involved solid phase extraction. Detection was performed using an Applied Biosystems Sciex API 2000 Mass spectrometer. The assay of lopinavir and ritonavir was linear over the range of 50 ng mL-1 to 20000 ng mL -1 and 20 ng mL -1 to 3000 ng mL-1 respectively with a precision of <15% and accuracy in the range of 85-115%. The limit of quantification in plasma for lopinavir and ritonavir was 50 ng mL -1 and 20 ng mL -1 respectively. The described method has the advantage of being rapid and easy and it could be applied in therapeutic monitoring of these drugs in human plasma

  8. Influence of estradiol, progesterone, and nutrition on concentrations of gonadotropins and GnRH receptors, and abundance of mRNA for GnRH receptors and gonadotropin subunits in pituitary glands of beef cows.

    Science.gov (United States)

    Looper, M L; Vizcarra, J A; Wettemann, R P; Malayer, J R; Braden, T D; Geisert, R D; Morgan, G L

    2003-01-01

    Nutritionally induced anovulatory cows (n = 28) were used to determine the effect of steroids on regulation of synthesis and secretion of gonadotropins. Anovulatory cows were ovariectomized and received intravaginal inserts containing estradiol (E2), progesterone (P4), E2 and P4 (E2P4), or a sham intravaginal insert (C) for 7 d. Concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were quantified in serum and E2 and P4 were quantified in plasma. Cows were exsanguinated within 1 to 2 h after removal of intravaginal inserts and pituitary glands were collected and stored at -80 degrees C until messenger ribonucleic acid (mRNA) for gonadotropin-releasing hormone receptor (GnRH-R) and gonadotropin subunits, pituitary content of GnRH-R, and LH and FSH were quantified. Pituitary glands from five proestrous cows were harvested to compare gonadotropin characteristics between ovariectomized, anovulatory cows and intact cows. Plasma concentrations of E2 were greater (P nutritionally induced anovulatory cows was increased (P nutritionally induced anovulatory cows with progesterone and estradiol may cause pulsatile secretion of LH.

  9. Bioadhesive emulsions for control release of progesterone resistant to vaginal fluids clearance.

    Science.gov (United States)

    Campaña-Seoane, Maria; Peleteiro, Aaron; Laguna, Reyes; Otero-Espinar, Francisco J

    2014-12-30

    The aim of this study is to propose that mucoadhesive vaginal emulsions can be able to resist the clearance effect of vaginal fluid and to have an effective control release of progesterone. With this goal, silicon derivative, cyclomethicone pentamer, was selected as the bioadhesive and water resistant material. In order to obtain a system which is insensitive to the dilution of aqueous fluids, water in silicone (W/S) emulsions were prepared and different proportions of cyclomethicone as well as 8% or 15% w/w of progesterone were employed. The rheological, mechanical and mucoadhesive properties of emulsions were characterized and the drug release was measured for each formulation. Mucoadhesive behavior was determined and the influence of simulated vaginal fluid (SVF) at bioadhesion was assessed using three commercial mucoadhesive vaginal gels (Crinone(®), K-Y jelly(®) and Zidoval(®)) as the bioadhesive references. All assayed emulsions have good rheological and mechanical properties and their consistence and viscosity increase when the proportion of the internal phase increases. Related to mucoadhesion, in the absence of SVF, W/S emulsions showed similar bioadhesive levels like the commercial formulations. However, in the presence of SVF, W/S emulsions are able to keep their mucoadhesive properties while the marketed references drastically lose their consistency and adherence to the vaginal mucosa. Drug release profiles from W/S emulsion show that progesterone is released with pseudo-order zero kinetics and a constant release rate is maintained for at least two weeks. The results of the in vivo studies developed in rats show that after a single vaginal administration, bioadhesive W/S emulsions increase the uterine tissue progesterone levels in young and postmenopausal rats. Moreover in postmenopausal rats, they provide high uterine levels of progesterone compared to the bioadhesive-marketed gel used as a reference. Therefore, W/S emulsions have an interesting

  10. Reduction of acid effects on trace element determination in food samples by CH4 mixed plasma-DRC-MS.

    Science.gov (United States)

    Guo, Wei; Hu, Shenghong; Zhang, Jiangyi; Hu, Zhaochu; Zhang, Hongfei; Wang, Yanxin

    2012-03-15

    A robust method for trace element determination in food samples by addition of methane to the plasma of a dynamic reaction cell mass spectrometer (CH(4) mixed plasma-DRC-MS) was developed. Addition of 3 mL min(-1) methane to Ar-plasma eliminates the signal suppressions of various elements (As, Se, Hg, etc.) due to the high concentration of nitric acid (10%, v/v). The CH(4)-Ar mixed plasma may compensate for the plasma cooling effects due to the highly concentrated nitric acid. The interfering polyatomic ions (40)Ar(12)C(+), (40)Ar(35)Cl(+) and (40)Ar(40)Ar(+) on (52)Cr(+), (75)As(+) and (80)Se(+) determination were removed effectively using the DRC with CH(4) as the reaction gas. The limits of quantification (LOQ, 10σ) were 0.35 ng g(-1), 0.07 ng g(-1), 0.35 ng g(-1), 0.07 ng g(-1), 0.15 ng g(-1), and 0.07 ng g(-1) for As, Cd, Cr, Hg, Pb and Se, respectively. The proposed method was applied to the determination of these trace elements in four food standard reference materials (NIST1577b, GBW10018, NIST1570a and GBW10016), and the results were in good agreement with the certified values.

  11. Spectrofluorimetric determination of sertraline in dosage forms and human plasma through derivatization with 9-fluorenylmethyl chloroformate

    Directory of Open Access Journals (Sweden)

    Belal Fathalla

    2011-10-01

    Full Text Available Abstract Background Sertraline is primarily used to treat major depression in adult outpatients as well as obsessive-compulsive, panic and social anxiety disorders in both adults and children. A survey of the literature reveals that most of the reported methods are either insufficiently sensitive or tedious and require highly sophisticated and dedicated instrumentation. The proposed method is considered to be specific for determination of SER in presence of its metabolite (deaminated form. Results A sensitive, simple and specific spectrofluorimetric method was developed for the determination of sertraline (SER in pharmaceutical formulations and biological fluids. The method is based on its reaction with 9-fluorenylmethyl chloroformate (FMOC-Cl in borate buffer of pH 8.0 to yield a highly fluorescent derivative peaking at 315 nm after excitation at 265 nm. The different experimental parameters affecting the development and stability of the reaction product were carefully studied and optimized. The fluorescence concentration plot was rectilinear over the range of 0.05-1.0 μg mL-1 with a lower detection limit of 5.34 × 10-3 μg mL-1 and limit of quantitation of 0.016 μg mL-1. Conclusions The proposed method was successfully applied to the analysis of commercial tablets and the results obtained were in good agreement with those obtained using the reference method. Furthermore, the method was applied for the determination of SER in spiked and real human plasma. The mean % recovery (n = 3 was 94.33 ± 1.53 and 92.00 ± 2.65, respectively. A proposal of the reaction pathway was postulated.

  12. Effect of two virus inactivation methods. Electron beam irradiation and binary ethylenimine treatment on determination of reproductive hormones in equine plasma

    Energy Technology Data Exchange (ETDEWEB)

    Kyvsgaard, N.C.; Nansen, P. [The Royal Veterinary and Agricultural Univ., Danish Centre for Experimental Parasitology, Frederiksberg (Denmark); Hoeier, R.; Brueck, I. [The Royal Veterinary and Agricultural Univ., Dept. of Clinical Studies, Section of Reproduction, Frederiksberg (Denmark)

    1997-12-31

    Ionizing irradiation and binary ethylenimine treatment have previously been shown to be effective for in-vitro inactivation of virus in biological material. In the present study the 2 methods were tested for possible effects on measurable concentrations of reproductive hormones in equine plasma (luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone (P{sub 4}), and oestradiol-17 {beta} (E{sub 2})). The inactivation methods were electron beam irradiation with a dose from 11 to 44 kGy or treatment with binary ethylenimine (BEI) in concentrations of 1 and 5 mmol/L. Generally, there was a close correlation (r>0.8, p<0.001) between pre- and post-treatment hormone levels. Thus, the different phases of the oestrous cycle could be distinguished on the basis of measured hormone concentrations of treated samples. However, both treatments significantly changed hormone concentrations of the plasma samples. For LH, FSH, and E{sub 2} the effect of irradiation and BEI treatment was depressive and dose-dependant. For P{sub 4} the effect of irradiation was also depressive and dose-dependant. However, the highest dose of BEI resulted in an increase of measured P{sub 4} concentration, which may be attributed to changes in the plasma matrix due to the treatment. Although the treatments affected measured hormone concentrations, the close correlation between pre-treatment and post-treatment measurements means that the diagnostic value will remain unchanged. (au). 17 refs.

  13. Experimental determination of the weld penetration evolution in keyhole plasma arc welding

    Institute of Scientific and Technical Information of China (English)

    Hu Qingxian; Wu Chuansong; Zhang Yuming

    2007-01-01

    Keyhole plasma arc welding experiments are conducted to measure the weld geometry and penetration at different moments during the initial phase from igniting arc to quasi-steady state. Indirect information on keyhole formation and evolution in plasma arc welding can be extracted based on the weld macrophotograph at cross section. It has laid foundation to verify the mathematical models of keyhole plasma arc welding.

  14. Precise timing for peak relaxin and decreased progesterone secretion after hysterectomy in the pig.

    Science.gov (United States)

    Felder, K J; Molina, J R; Benoit, A M; Anderson, L L

    1986-10-01

    Relaxin and progesterone secretion by aging corpora lutea (days 90-120) was examined in pregnant and lactating gilts compared with that in hysterectomized animals. The length of pregnancy is about 115 days in pigs. Unmated gilts were hysterectomized on day 6 (estrus = day 0). From days 90-101, relaxin concentrations in peripheral plasma remained consistently low in pregnant gilts (range, 0.7-1.5 ng/ml) and less (P less than 0.05) than those in hysterectomized animals (range, 0.9-3.5 ng/ml). Relaxin increased abruptly (P less than 0.01) to a peak of 66 ng/ml in pregnant gilts and 37 ng/ml in hysterectomized animals. Relaxin peaked in pregnant animals at 113 +/- 0.7 days (+/- SE) and in hysterectomized gilts at 113 +/- 0.7 days; gestation length averaged 114 +/- 0.8 days. In pregnant gilts, relaxin decreased from a peak of 66 to 11 ng/ml within 1 day and remained low (less than 1.0 ng/ml) in these lactating dams until day 120. In hysterectomized gilts, peak relaxin also decreased abruptly from 37 to 4.2 ng/ml, but remained consistently greater (P less than 0.05) than that in lactating dams. Although there were abrupt shifts in relaxin concentrations within 20 min, there was no evidence for consistent episodic relaxin release between days 112-116. Plasma progesterone concentrations were consistently greater (P less than 0.05) in hysterectomized than in pregnant gilts from days 102-110. Progesterone decreased abruptly in prepartum gilts (days 111-114) from 16 to 1.2 ng/ml and remained low during lactation (0.5 ng/ml). In hysterectomized animals, it decreased abruptly on days 110-113, ranging from 20-12 ng/ml, and remained at this lower level until day 120. These results clearly indicate that a precisely timed peak release of relaxin and coincident decrease in progesterone secretion occur in unmated hysterectomized gilts at the same time as those found a few hours preceding parturition during normal pregnancy. These abrupt shifts in relaxin and progesterone secretion on

  15. Individual and combined effects of relaxin, estrogen, and progesterone in ovariectomized gilts. I. Effects on the growth, softening, and histological properties of the cervix.

    Science.gov (United States)

    Winn, R J; Baker, M D; Sherwood, O D

    1994-09-01

    Marked growth and softening of the uterine portion of the cervix occur during the last third of the 115-day gestation period in the gilt. These changes in the cervix are temporally correlated with elevated blood levels of relaxin, estrogen, and progesterone. We recently demonstrated that relaxin plays a major role in promoting both the growth and softening of the cervix that occur in pregnant gilts. The roles of estrogen and progesterone in these cervical changes remain poorly understood. Accordingly, this study determined the influence of relaxin, estrogen, and progesterone, individually and in combination, on cervical growth and softening in gilts. Fifteen days after ovariectomy, six to nine nonpregnant, sexually mature gilts were assigned to one of the following eight treatment groups: ovariectomized controls, relaxin treated, estrogen treated, progesterone treated, estrogen plus relaxin treated, progesterone plus relaxin treated, estrogen plus progesterone treated, and progesterone plus estrogen plus relaxin treated. Treatment was given for 10 days, with doses of relaxin (0.5 mg, four times daily), estradiol benzoate (1 mg, twice daily), and progesterone (50 mg, twice daily) selected to provide blood levels resembling those between days 100-110 of gestation. The growth, softening, and histological characteristics of the cervices were determined. Treatment with relaxin significantly increased the growth and softening and altered the histological characteristics of the uterine portion of the cervix in the absence of steroid treatment. Estrogen treatment alone increased cervical growth, but when given in combination with relaxin, estrogen did not augment relaxin's ability to increase either cervical growth or softening. Progesterone alone had little or no effect on the growth or softening of the uterine portion of the cervix. Unexpectedly, when given in combination with relaxin, progesterone augmented markedly relaxin's effects on softening and alteration of the

  16. Increased doses of vaginal progesterone for the prevention of preterm birth in twin pregnancies: a randomised controlled double-blind multicentre trial.

    Science.gov (United States)

    Serra, V; Perales, A; Meseguer, J; Parrilla, J J; Lara, C; Bellver, J; Grifol, R; Alcover, I; Sala, M; Martínez-Escoriza, J C; Pellicer, A

    2013-01-01

    Previous trials have shown little benefit for preventing preterm birth in twin pregnancies using 90-200 mg of daily vaginal natural progesterone. Higher doses have not been tested. Our aim was to determine the efficacy and safety of two different daily doses of vaginal natural progesterone (200 and 400 mg), compared with placebo, for preventing preterm birth in unselected twin pregnancies. Randomised controlled double-blind multicentre trial (1:1:1). The study was carried out in five university centres from Valencia, Murcia and Alicante (Spain). Women with dichorionic diamniotic twin pregnancies. The women self-inserted two vaginal pessaries daily, containing placebo (n = 96), 200 mg of natural progesterone (n = 97) or 400 mg of natural progesterone (n = 97), from 20 to 34 weeks of gestation or delivery. Randomisation was performed by an external centre. Data were analysed on an intention-to-treat basis. Preterm birth rate. The baseline characteristics for placebo and progesterone groups were similar. Comparison of the three groups and analysis of progesterone-treated versus untreated women showed similar pregnancy and neonatal outcomes. The proportion of preterm and very preterm births, low birthweight, perinatal mortality and neonatal morbidity showed no differences between the three groups. Similar results were also obtained when comparing the 200- versus 400-mg progesterone groups. No serious adverse effects were encountered. Vaginal progesterone therapy was generally well tolerated, but failed to prevent preterm births in unselected dichorionic diamniotic twin pregnancies. The 400-mg progesterone dose offered no advantages over the 200-mg regimen. © 2012 The Authors BJOG An International Journal of Obstetrics and Gynaecology © 2012 RCOG.

  17. The rapid immunosuppression in phytohemagglutinin-activated human T cells is inhibited by the proliferative Ca(2+) influx induced by progesterone and analogs.

    Science.gov (United States)

    Lin, Veronica Hui-Chen; Chen, Jiann-Jong; Liao, Chen-Chung; Lee, Shinn-Shing; Chien, Eileen Jea

    2016-07-01

    Progesterone, an endogenous immunomodulator, suppresses human T-cell activation during pregnancy. A sustained Ca(2 +) influx is an important signal for T-cell proliferation after crosslinking of T-cell receptor/CD3 complexes by anti-CD3 antibodies or phytohemagglutinin (PHA). Progesterone targets cell membrane sites inducing rapid responses including elevated intracellular free calcium concentration ([Ca(2+)]i) and suppressed T-cell PHA-activated proliferation. Interestingly, both PHA and progesterone induce [Ca(2+)]i elevation, but it remains unclear whether the PHA-induced Ca(2+) influx is affected by progesterone leading to T-cell immunosuppression. Primary T-cells were isolated from human peripheral blood and the quench effect on intracellular fura-2 fluorescence of Mn(2+) was used to explore the responses to Ca(2+) influx with cell proliferation being determined by MTT assay. PHA-stimulated Ca(2+) influx was dose-dependently suppressed by progesterone and its agonist R5020, which correlated with PHA-activated T-cell proliferation inhibition. A similar dose-dependent suppression effect on cellular Ca(2+) influx and proliferation occurred with the TRPC channel inhibitor BTP2 and selective TRPC3 channel inhibitor Pyr3. In addition, two progesterone analogs, Org OD 02-0 and 20α-hydroxyprogesterone (20α-OHP), also produced dose-dependent suppression of Ca(2+) influx, but had no effect on proliferation. Finally, inhibition of PHA-activated T-cell proliferation by progesterone is further suppressed by 20α-OHP, but not by Org OD 02-0. Overall, progesterone and R5020 are able to rapidly decrease PHA-stimulated sustained Ca(2+) influx, probably via blockade of TRPC3 channels, which suppresses T-cell proliferation. Taken together, the roles of progesterone and its analogs regarding the rapid response Ca(2+) influx need to be further explored in relation to cytokine secretion and proliferation in activated T-cells.

  18. Preparation and preliminary bioevaluation of {sup 99m}Tc(CO){sub 3}-11{beta}-progesterone derivative prepared via click chemistry route

    Energy Technology Data Exchange (ETDEWEB)

    Dhyani, Manish V.; Satpati, Drishty; Korde, Aruna [Radiopharmaceuticals Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India); Sarma, Haladhar Dev [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India); Kumar, Chandan [Radiopharmaceuticals Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India); Banerjee, Sharmila, E-mail: sharmila@barc.gov.i [Radiopharmaceuticals Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India)

    2010-11-15

    Introduction: Progesterone receptors (PRs) overexpressed in breast cancers serve as potential targets for developing radiotracers for use in nuclear medicine. Hence, suitably derivatized progesterone can be envisaged as a potential vector for targeting overexpression of receptors in breast cancer. In the present article, we report the preparation of a {sup 99m}Tc(CO){sub 3}-progesterone triazole using the Cu(I)-catalyzed novel click chemistry route. Preliminary evaluation of the radiolabeled derivative has been carried out in binding studies with MCF 7 cell lines. Methods: 11-Hydroxyprogesterone has been synthetically derivatized to 11-azidoprogesterone. Subsequently, the cycloaddition reaction between progesterone azide and propargyl glycine was carried out to prepare 1,4-bifunctionalized progesterone triazole analogue. The clicked progesterone triazole derivative was radiolabeled with {sup 99m}Tc and characterized by HPLC. The chemical characterization of {sup 99m}Tc(CO){sub 3}-progesterone triazole has been carried out by preparing its corresponding rhenium complex using the [NEt{sub 4}]{sub 2}[Re(CO){sub 3}Br{sub 3}] precursor. While in vitro studies were carried out in MCF7 cell lines, in vivo distribution studies were performed in female Swiss mice. Results: The radiolabeled complex could be prepared in >95% radiochemical yield as determined by HPLC. In vitro studies of {sup 99m}Tc(CO){sub 3}-progesterone complex in MCF7 cell lines overexpressing receptors for breast cancer showed binding up to 30%. In vivo distribution studies in female Swiss mice have shown uterine uptake of 0.41 (0.06) % ID/g at 3 h postinjection (pi) and retention therein till 24 h pi. Conclusion: The present study demonstrates a novel and facile route for preparation of {sup 99m}Tc-labeled progesterone complex using click chemistry. This strategy can be further extended towards preparation of radiolabeled complexes of other steroidal derivatives.

  19. Validation of a simple gas chromatographic-mass spectrometric method for the determination of gamma-butyrolactone in human plasma.

    Science.gov (United States)

    Fukui, Yousuke; Matsusima, Eiji; Muramoto, Kouichi; Nagai, Nobutaka; Ohama, Koso; Yamashita, Kazumasa

    2003-02-25

    A gas chromatographic-mass spectrometric (GC-MS) method is described for the determination of human plasma levels of gamma-butyrolactone (GBL) is described. The method is sensitive and simple. The plasma sample spiked with the internal standard was extracted by dichloromethane (CH(2)Cl(2)) in acidic conditions, and the concentrated organic layer was injected into GC-MS. Because of endogenous GBL in human plasma, the method used a standard calibration curve. The calibration curve was linear from 10 to 1000 ng/ml. The method has been validated for accuracy and precision with the relative error and C.V. for intra- and inter-day within 10%. GBL-spiked plasma samples stored at -80 degrees C were stable for a 3-month period. The stability of plasma samples after three cycles of freezing and thawing and of prepared samples on an autosampler for 48 h were demonstrated. Plasma concentrations of GBL before and after administration of UFT were 24.3+/-14.2 and 84.9+/-22.4 ng/ml, respectively.

  20. A Validated HPLC-DAD Method for Simultaneous Determination of Etodolac and Pantoprazole in Rat Plasma

    Directory of Open Access Journals (Sweden)

    Ali S. Abdelhameed

    2014-01-01

    Full Text Available A simple, sensitive, and accurate HPLC-DAD method has been developed and validated for the simultaneous determination of pantoprazole and etodolac in rat plasma as a tool for therapeutic drug monitoring. Optimal chromatographic separation of the analytes was achieved on a Waters Symmetry C18 column using a mobile phase that consisted of phosphate buffer pH~4.0 as eluent A and acetonitrile as eluent B in a ratio of A : B, 55 : 45 v/v for 6 min, pumped isocratically at a flow rate of 0.8 mL min−1. The eluted analytes were monitored using photodiode array detector set to quantify samples at 254 nm. The method was linear with r2=0.9999 for PTZ and r2=0.9995 for ETD at a concentration range of 0.1–15 and 5–50 μgmL−1 for PTZ and ETD, respectively. The limits of detection were found to be 0.033 and 0.918 μgmL−1 for PTZ and ETD, respectively. The method was statistically validated for linearity, accuracy, precision, and selectivity following the International Conference for Harmonization (ICH guidelines. The reproducibility of the method was reliable with the intra- and interday precision (% RSD <7.76% for PTZ and <7.58 % for ETD.

  1. Metal and metalloid determination in bioethanol through inductively coupled plasma-optical emission spectroscopy

    Science.gov (United States)

    Sánchez, Carlos; Lienemann, Charles-Philippe; Todolí, José-Luis

    2016-01-01

    A new method to carry out the elemental determination of metals in bioethanol through ICP-OES has been developed. The procedure is based on the use of a heated torch integrated sample introduction system (hTISIS) to directly introduce the vaporized sample into the plasma. Two injection modes (continuous liquid aspiration and air-segmented flow injection analysis) have been evaluated. In a first step, the matrix effects caused by several ethanol-water mixtures were removed by operating the hTISIS at 400 °C in segmented injection. Meanwhile, the results also proved that the system could be operated in continuous mode at 200 °C with the complete interference removal. Finally, twenty-eight real samples with bioethanol contents between 55% and 100% were analyzed with the methods previously developed. Regarding validation, recoveries from 80% to 120% were obtained for 18 analytes and the concentrations found with the proposed method were in agreement with those encountered with a preconcentration method, taken as a reference procedure. Limits of detection went from 3 ng mL- 1 for manganese to about 500 ng mL- 1 for calcium. This allowed to quantify Cr, Fe, Mg, Mn and Zn in segmented flow injection and Al, Cd, Cr, Cu, K, Mg, Mn, Na and Zn in continuous sample aspiration mode in bioethanol samples.

  2. Determination of Menthol in Plasma and Urine by Gas Chromatography/Mass Spectrometry (GC/MS).

    Science.gov (United States)

    Peat, Judy; Frazee, Clint; Kearns, Gregory; Garg, Uttam

    2016-01-01

    Menthol, a monoterpene, is a principal component of peppermint oil and is used extensively in consumer products as a flavoring aid. It is also commonly used medicinally as a topical skin coolant; to treat inflammation of the mucous membranes, digestive problems, and irritable bowel syndrome (IBS); and in preventing spasms during endoscopy and for its spasmolytic effect on the smooth muscle of the gastrointestinal tract. Menthol has a half life of 3-6 h and is rapidly metabolized to menthol glucuronide which is detectable in urine and serum following menthol use. We describe a method for the determination of total menthol in human plasma and urine using liquid/liquid extraction, gas chromatography/mass spectrometry (GC/MS) in selected ion monitoring mode and menthol-d4 as the internal standard. Controls are prepared with menthol glucuronide and all samples undergo enzymatic hydrolysis for the quantification of total menthol. The method has a linear range of 5-1000 ng/mL, and coefficient of variation <10%.

  3. Determination of impurity elements in MnZn ferrites by inductively coupled plasma mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    张萍; 符靓; 马俊才; 唐有根

    2015-01-01

    An inductively coupled plasma mass spectrometry (ICP-MS) method was developed for the determination of Na, Mg, Al, K, Ca, Ti, Cr, Co, Ni, Cu, Ga, As, Mo, Ag, Cd and Pb in MnZn ferrites. The sample was digested by HNO3+HCl with microwave digestion followed by dilution with ultrapure water, then the above 16 impurity elements in the solution were analyzed directly by ICP-MS. The impurity elements were introduced by the helium gas or hydrogen gas into the octopole reaction system (ORS) to eliminate the polyatomic interferences caused by the high salty matrixes. The matrix effect was minimized through matrix matching, and Be, Y and Rh were used as internal standard elements. The working parameters of the instrument were optimized. The results show that the method has good precision and high accuracy. The detection limits for the investigated elements are in the range of 0.9−37.5 ng/L, the relative standard deviation of each element is within 1.1%−4.8%, and the recovery of each element is 90%−108%.

  4. Experimental Determination of DT Yield in High Current DD Dense Plasma Focii

    Energy Technology Data Exchange (ETDEWEB)

    Lowe, D. R. [National Security Technologies, LLC; Hagen, E. C. [National Security Technologies, LLC; Meehan, B. T. [National Security Technologies, LLC; Springs, R. K. [University of Nevada, Las Vegas; O' Brien, R. J. [University of Nevada, Las Vegas

    2013-06-18

    Dense Plasma Focii (DPF), which utilize deuterium gas to produce 2.45 MeV neutrons, may in fact also produce DT fusion neutrons at 14.1 MeV due to the triton production in the DD reaction. If beam-target fusion is the primary producer of fusion neutrons in DPFs, it is possible that ejected tritons from the first pinch will interact with the second pinch, and so forth. The 2 MJ DPF at National Security Technologies’ Losee Road Facility is able to, and has produced, over 1E12 DD neutrons per pulse, allowing an accurate measurement of the DT/DD ratio. The DT/DD ratio was experimentally verified by using the (n,2n) reaction in a large piece of praseodymium metal, which has a threshold reaction of 8 MeV, and is widely used as a DT yield measurement system1. The DT/DD ratio was experimentally determined for over 100 shots, and then compared to independent variables such as tube pressure, number of pinches per shot, total current, pinch current and charge voltage.

  5. Determination of stable cesium and strontium in rice samples by inductively coupled plasma mass spectrometry

    Science.gov (United States)

    Srinuttrakul, W.; Yoshida, S.

    2017-06-01

    For long-term radiation dose assessment models, food ingestion is one of the major exposure pathways to human. In general, the stable isotopes can serve as analogues of radioisotopes. In this study, rice samples were collected from 30 paddy fields in Si Sa Ket, Yasothon and Roi Et in the northeast of Thailand in November 2014. The concentrations of stable cesium (Cs-133) and strontium (Sr-88) in polished rice were determined by inductively coupled plasma mass spectrometry (ICP-MS). The standard reference material of rice flour (NIST 1568a) with spiked Cs and Sr was used to validate the analytical method. The concentration of Cs in polished rice from Si Sa Ket, Yasothon and Roi Et was 0.158 ± 0.167 mg kg-1, 0.090 ± 0.117 mg kg-1 and 0.054 ± 0.031 mg kg-1, respectively. The concentration of Sr in polished rice from Si Sa Ket, Yasothon and Roi Et was 0.351 ± 0.108 mg kg-1, 0.364 ± 0.215 mg kg-1 and 0.287 ± 0.102 mg kg-1, respectively. Comparison of the results with Japanese data before the Fukushima Di-ichi nuclear power plant accident showed that the concentrations of both Cs and Sr for Thai rice were higher than those for Japanese rice.

  6. Determination of cobalamins using capillary electrophoresis inductively coupled plasma mass spectrometry

    Science.gov (United States)

    Baker, S. A.; Miller-Ihli, N. J.

    2000-12-01

    The determination of cobalamins using capillary electrophoresis inductively coupled plasma mass spectrometry (CE-ICP-MS) was investigated. Both capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) modes of operation were studied. The optimal separation of four cobalamin species (cyanocobalamin, hydroxocobalamin, methylcobalamin, and 5'-deoxyadenosylcobalamin) and a potentially harmful corrinoid analogue (cobinamide dicyanide) was obtained using CZE at a pH of 2.5. Both 20 mM phosphate and 20 mM formate buffers were used with success, although the formate buffer provided improved resolution. The CZE-ICP-MS method was used to quantify cyanocobalamin in a vitamin supplement and the analytical results were in good agreement (±5%) with values obtained by ICP-MS for total Co levels. The solution detection limits for cobalamins using CZE-ICP-MS were approximately 50 ng/ml. MEKC was found to be useful for the screening of vitamin preparations because it provided a rapid means of distinguishing cyanocobalamin (the form most commonly used in vitamin preparations) from free cobalt. The separation of free cobalt and cyanocobalamin using MEKC was achieved in less than 10 min.

  7. Determination of omeprazole and its metabolites in human plasma by liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Kanazawa, Hideko; Okada, Akiko; Matsushima, Yoshikazu; Yokota, Hiromitsu; Okubo, Shigeo; Mashige, Fumiko; Nakahara, Kazuhiko

    2002-03-08

    Omeprazole is a benzimidazole compound that acts as a proton-pump inhibitor. Because the metabolism of omeprazole is mainly catalyzed by cytochrome P-450 (CYP) 3A4 and CYP2C19. the genetic polymorphism of CYP2C19 could be of clinical concern in the treatment of acid-related diseases with omeprazole. Therefore, a reliable method for omeprazole phenotyping is desirable in clinical situations. This study has demonstrated the determination of omeprazole and its metabolites in human plasma by liquid chromatography-three-dimensional quadrupole mass spectrometry with a sonic spray ionization interface. The analytical column was YMC-Pack Pro C18(50x2.0 mm I.D.) using acetonitrile-50 mM ammonium acetate (pH 7.25) (1:4) at a flow-rate of 0.2 ml/min. The drift voltage was 30 V. The sampling aperture was heated at 110 degrees C and Shield temperature was 230 degrees C. In the mass spectrum, the molecular ions of omeprazole, hydroxyomeprazole and omeprazole sulfone were clearly observed as base peaks. This method is sufficiently sensitive and accurate for pharmacokinetic studies of omeprazol.

  8. In situ determination of the static inductance and resistance of a plasma focus capacitor bank.

    Science.gov (United States)

    Saw, S H; Lee, S; Roy, F; Chong, P L; Vengadeswaran, V; Sidik, A S M; Leong, Y W; Singh, A

    2010-05-01

    The static (unloaded) electrical parameters of a capacitor bank are of utmost importance for the purpose of modeling the system as a whole when the capacitor bank is discharged into its dynamic electromagnetic load. Using a physical short circuit across the electromagnetic load is usually technically difficult and is unnecessary. The discharge can be operated at the highest pressure permissible in order to minimize current sheet motion, thus simulating zero dynamic load, to enable bank parameters, static inductance L(0), and resistance r(0) to be obtained using lightly damped sinusoid equations given the bank capacitance C(0). However, for a plasma focus, even at the highest permissible pressure it is found that there is significant residual motion, so that the assumption of a zero dynamic load introduces unacceptable errors into the determination of the circuit parameters. To overcome this problem, the Lee model code is used to fit the computed current trace to the measured current waveform. Hence the dynamics is incorporated into the solution and the capacitor bank parameters are computed using the Lee model code, and more accurate static bank parameters are obtained.

  9. Determination of marbofloxacin in plasma and synovial fluid by ultrafiltration followed by HPLC-MS/MS.

    Science.gov (United States)

    Montesano, Camilla; Curini, Roberta; Sergi, Manuel; Compagnone, Dario; Celani, Gianluca; Varasano, Vincenzo; Petrizzi, Lucio; Amorena, Michele

    2016-05-10

    A rapid LC-MS/MS method for the determination of marbofloxacin in plasma and synovial fluid is presented in this study. The method uses a rapid sample preparation which only requires an ultrafiltration step with centrifugal filter devices. The optimized procedure allows a minimal need of sample (175 μL), particularly useful for synovial fluid samples which amount is rather limited; it is simple, rapid and easily applicable providing anyhow a satisfactory clean up, demonstrated by post-infusion experiments. On the other hand to maximize the speed of the analysis an ultrafast chromatographic separation has been obtained by selecting a column of 20 mm; the reduced run-time is suitable for processing numerous samples on a daily basis. Linearity was assessed in the range 5-2500 ng mL(-1); ofloxacin was used as internal standard. LOD and LOQ were respectively 1 and 5 ng/mL. The method was successfully applied to a set of samples generated during an experimental veterinary study.

  10. Simultaneous determination of clobazam and its major metabolite in human plasma by a rapid HPLC method.

    Science.gov (United States)

    Rouini, Mohammadreza; Ardakani, Yalda H; Hakemi, Lida; Mokhberi, Maryam; Badri, Gheise

    2005-09-05

    A rapid and specific HPLC method has been developed and validated for simultaneous determination of clobazam, the anticonvulsant agent, and its major metabolite in human plasma. The sample preparation was a liquid-liquid extraction with tuloene yielding almost near 100% recoveries of two compounds. Chromatographic separation was achieved with a Chromolith Performance RP-18e 100 mm x 4.6mm column, using a mixture of a phosphate buffer (pH 3.5; 10mM)-acetonitrile (70:30, v/v), in isocratic mode at 2 ml/min at a detection wave-length of 228 nm. The calibration curves were linear (r(2)>0.998) in the concentration range of 5-450 ng ml(-1). The lower limit of quantification was 5 ng ml(-1) for two compounds studied. The within- and between-day precisions in the measurement of QC samples at four tested concentrations were in the range of 0.89-9.1% and 2.1-10.1% R.S.D., respectively. The developed procedure was applied to assess the pharmacokinetics of clobazam and its major metabolite following administration of a single 10mg oral dose of clobazam to healthy volunteers.

  11. Determination of piracetam in rat plasma by LC-MS/MS and its application to pharmacokinetics.

    Science.gov (United States)

    Wang, Xianqin; Zhu, Jiayin; Xu, Renai; Yang, Xuezhi; Wu, Haiya; Lin, Dan; Ye, Faqing; Hu, Lufeng

    2010-10-01

    A sensitive and selective liquid chromatography-tandem mass spectrometry method for the determination of piracetam in rat plasma was developed and validated over the concentration range of 0.1-20 µg/mL. After addition of oxiracetam as internal standard, a simplified protein precipitation with trichloroacetic acid (5%) was employed for the sample preparation. Chromatographic separation was performed by a Zorbax SB-Aq column (150 × 2.1 mm, 3.5 µm). The mobile phase was acetonitrile-1% formic acid in water (10:90 v/v) delivered at a flow rate of 0.3 mL/min. The MS data acquisition was accomplished in multiple reaction monitoring mode with a positive electrospray ionization interface. The lower limit of quantification was 0.1 µg/mL. For inter-day and intra-day tests, the precision (RSD) for the entire validation was less than 9%, and the accuracy was within the 94.6-103.2% range. The developed method was successfully applied to pharmacokinetic studies of piracetam in rats following single oral administration dose of 50 mg/kg. Copyright © 2010 John Wiley & Sons, Ltd.

  12. Determination of memantine in plasma and vitreous humour by HPLC with precolumn derivatization and fluorescence detection.

    Science.gov (United States)

    Puente, Belen; Hernandez, Esther; Perez, Susana; Pablo, Luis; Prieto, Esther; Garcia, Maria Angeles; Bregante, Miguel Angel

    2011-01-01

    A new HPLC procedure with precolumn derivatization and rimantadine as the internal standard for determining memantine, a candidate agent for the treatment of glaucoma in plasma and vitreous humour, has been developed and validated. Precolumn derivatization was performed with 9-fluorenylmethyl-chloroformate-chloride (FMOC-Cl) as the derivatization reagent and followed by a liquid-liquid extraction with n-hexane. Optimal conditions for derivatization were an FMOC-Cl concentration of 1.5 mM, a reaction time of 20 min, the temperature at 30°C, the borate buffer pH 8.5, and a borate buffer-acetonitrile ratio of 1:1. The derivatives were analyzed by isocratic HPLC with the fluorescence detector λex 260 nm λem 315 nm on a Novapack C(18) reversed-phase column with a mobile phase of acetonitrile-water (73:27, v/v), 40°C, and a flow rate of 1.2 mL/min. The linear range was 10-1000 ng/mL with a quantification limit of ∼ 10 ng/mL for both types of samples. This analytical method may be suitable for using in ocular availability studies.

  13. Determination of Dynamics of Plant Plasma Membrane Proteins with Fluorescence Recovery and Raster Image Correlation Spectroscopy.

    Science.gov (United States)

    Laňková, Martina; Humpolíčková, Jana; Vosolsobě, Stanislav; Cit, Zdeněk; Lacek, Jozef; Čovan, Martin; Čovanová, Milada; Hof, Martin; Petrášek, Jan

    2016-04-01

    A number of fluorescence microscopy techniques are described to study dynamics of fluorescently labeled proteins, lipids, nucleic acids, and whole organelles. However, for studies of plant plasma membrane (PM) proteins, the number of these techniques is still limited because of the high complexity of processes that determine the dynamics of PM proteins and the existence of cell wall. Here, we report on the usage of raster image correlation spectroscopy (RICS) for studies of integral PM proteins in suspension-cultured tobacco cells and show its potential in comparison with the more widely used fluorescence recovery after photobleaching method. For RICS, a set of microscopy images is obtained by single-photon confocal laser scanning microscopy (CLSM). Fluorescence fluctuations are subsequently correlated between individual pixels and the information on protein mobility are extracted using a model that considers processes generating the fluctuations such as diffusion and chemical binding reactions. As we show here using an example of two integral PM transporters of the plant hormone auxin, RICS uncovered their distinct short-distance lateral mobility within the PM that is dependent on cytoskeleton and sterol composition of the PM. RICS, which is routinely accessible on modern CLSM instruments, thus represents a valuable approach for studies of dynamics of PM proteins in plants.

  14. Evaluation of an immunoassay for determination of plasma efavirenz concentrations in resource-limited settings

    DEFF Research Database (Denmark)

    Abdissa, Alemseged; Wiesner, Lubbe; McIlleron, Helen

    2014-01-01

    to be implemented in resource-limited settings. This study evaluated a commercially available immunoassay for measurement of plasma efavirenz. Methods: The immunoassay-based method was applied to measure efavirenz using a readily available Humastar 80 chemistry analyzer. We compared plasma efavirenz concentrations...

  15. Determination of lidocaine in plasma by direct solid-phase microextraction combined with gas chromatography

    NARCIS (Netherlands)

    Koster, EHM; Wemes, C; Morsink, JB; de Jong, GJ

    2000-01-01

    Direct-immersion solid-phase microextraction (SPME) has been used to extract the local anesthetic lidocaine from human plasma. A simplified model shows the relationship between the total amount of drug in plasma and the amount of drug extracted. The model takes into account that the drug participate

  16. Collective Thomson scattering system for determination of ion properties in a high flux plasma beam

    NARCIS (Netherlands)

    van der Meiden, H. J.; Vernimmen, J. W. M.; Bystrov, K.; Jesko, K.; Kantor, M. Y.; De Temmerman, G.; Morgan, T. W.

    2016-01-01

    A collective Thomson scattering system has been developed for measuring ion temperature, plasma velocity and impurity concentration in the high density magnetized Magnum-PSI plasma beam, allowing for measurements at low temperature (<5 eV) and high electron density >4 × 1020 m−3, while

  17. Determination of platinum surface contamination in veterinary and human oncology centres using inductively coupled plasma mass spectrometry

    NARCIS (Netherlands)

    Janssens, T.; Brouwers, E. E M; de Vos, J. P.; de Vries, N.; Schellens, J. H M; Beijnen, J. H.

    2015-01-01

    The objective of this study was to determine the surface contamination with platinum-containing antineoplastic drugs in veterinary and human oncology centres. Inductively coupled plasma mass spectrometry was used to measure platinum levels in surface samples. In veterinary and human oncology

  18. IFCC reference measurement procedure for substance concentration determination of total carbon dioxide in blood, plasma or serum

    NARCIS (Netherlands)

    Burnett, RW; Covington, AK; Fogh-Andersen, N; Kulpmann, WR; Lewenstam, A; Mas, AHJ; VanKessel, AL; Zijlstra, WG

    2001-01-01

    A reference measurement procedure for substance concentration determination of total CO, in blood, plasma (the anticoagulant is usually heparin) or serum is described. The document covers the principle of the method, the materials and equipment needed and essential aspects of the procedure. The subs

  19. Rapid and simple clean-up and derivatizaton procedure for the gas chromatographic determination of acidic drugs in plasma

    NARCIS (Netherlands)

    Roseboom, H.; Hulshoff, A.

    1979-01-01

    A rapid and simple clean-up and derivatization procedure that can be generally applied to the gas chromatographie (GC) determination of acidic drugs of various chemical and therapeutic classes is described. The drugs are extracted from acidified plasma with chloroform containing 5% of isopropanol, w

  20. Copper Determination in Gunshot Residue by Cyclic Voltammetric and Inductive Coupled Plasma-Optical Emission Spectroscopy

    Directory of Open Access Journals (Sweden)

    Mohd Hashim Nurul’Afiqah Hashimah

    2016-01-01

    Full Text Available Analysis of gunshot residue (GSR is a crucial evidences for a forensic analyst in the fastest way. GSR analysis insists a suitable method provides a relatively simple, rapid and precise information on the spot at the crime scene. Therefore, the analysis of Cu(II in GSR using cyclic voltammetry (CV on screen printed carbon electrode (SPCE is a better choice compared to previous alternative methods such as Inductive Coupled Plasma-Optical Emission Spectroscopy (ICP-OES those required a long time for analysis. SPCE is specially designed to handle with microvolumes of sample such as GSR sample. It gives advantages for identification of copper in GSR on-site preliminary test to prevent the sample loss on the process to be analyzed in the laboratory. SPCE was swabbed directly on the shooter’s arm immediately after firing and acetate buffer was dropped on SPCE before CV analysis. For ICP-OES analysis, cotton that had been soaked in 0.5 M nitric acid was swabbed on the shooter’s arm immediately after firing and kept in a tightly closed sampling tube. Gold coated SPCE that had been through nanoparticles modification exhibits excellent performance on voltammograms. The calibration was linear from 1 to 50 ppm of copper, the limit of detection for copper was 0.3 ppm and a relative standard deviation was 6.1 %. The method was successfully applied to the determination of copper in GSR. The Cu determination on SPCE was compared and validated by ICP-OES method with 94 % accuracy.

  1. Determination of iodine and bromine compounds in foodstuffs by CE-inductively coupled plasma MS.

    Science.gov (United States)

    Chen, Jing-Huan; Wang, Kai-en; Jiang, Shiuh-Jen

    2007-11-01

    A CE-inductively coupled plasma mass spectrometric (CE-ICP-MS) method for iodine and bromine speciation analysis is described. Samples containing ionic iodine (I(-) and IO(3)(-)) and bromine (Br(-) and BrO(3)(-)) species are subjected to electrophoretic separation before injection into the microconcentric nebulizer (CEI-100). The separation has been achieved in a 50 cm length x 75 microm id fused-silica capillary. The electrophoretic buffer used is 10 mmol/L Tris (pH 8.0), while the applied voltage is set at -8 kV. Detection limits are 1 and 20-50 ng/mL for various I and Br compounds, respectively, based on peak height. The RSD of the peak areas for seven injections of 0.1 microg/mL I(-), IO(3)(-) and 1 microg/mL Br(-), BrO(3)(-) mixture is in the range of 3-5%. This method has been applied to determine various iodine and bromine species in NIST SRM 1573a Tomato Leaves reference material and a salt and seaweed samples obtained locally. A microwave-assisted extraction method is used for the extraction of these compounds. Over 87% of the total iodine and 83% of the total bromine are extracted using a 10% m/v tetramethylammonium hydroxide (TMAH) solution in a focused microwave field within a period of 10 min. The spike recoveries are in the range of 94-105% for all the determinations. The major species of iodine and bromine in tomato leaves, salt, and seaweed are Br(-), IO(3)(-), I(-), and Br(-), respectively.

  2. The effect of sample size on fresh plasma thromboplastin ISI determination

    DEFF Research Database (Denmark)

    Poller, L; Van Den Besselaar, A M; Jespersen, J;

    1999-01-01

    The possibility of reduction of numbers of fresh coumarin and normal plasmas has been studied in a multicentre manual prothrombin (PT) calibration of high international sensitivity index (ISI) rabbit and low ISI human reference thromboplastins at 14 laboratories. The number of calibrant plasmas...... was reduced progressively by a computer program which generated random numbers to provide 1000 different selections for each reduced sample at each participant laboratory. Results were compared with those of the full set of 20 normal and 60 coumarin plasma calibrations. With the human reagent, 20 coumarins...... and seven normals still achieved the W.H.O. precision limit (3% CV of the slope), but with the rabbit reagent reduction coumarins with 17 normal plasmas led to unacceptable CV. Little reduction of numbers from the full set of 80 fresh plasmas appears advisable. For maximum confidence, when calibrating...

  3. Unbound fraction of fluconazole and linezolid in human plasma as determined by ultrafiltration: Impact of membrane type.

    Science.gov (United States)

    Kratzer, Alexander; Kees, Frieder; Dorn, Christoph

    2016-12-15

    Ultrafiltration is a rapid and convenient method to determine the free concentrations of drugs in plasma. Several ultrafiltration devices based on Eppendorf cups are commercially available, but are not validated for such use by the manufacturer. Plasma pH, temperature and relative centrifugal force as well as membrane type can influence the results. In the present work, we developed an ultrafiltration method in order to determine the free concentrations of linezolid or fluconazole, both neutral and moderately lipophilic antiinfective drugs for parenteral as well as oral administration, in plasma of patients. Whereas both substances behaved relatively insensitive in human plasma regarding variations in pH (7.0-8.5), temperature (5-37°C) or relative centrifugal force (1000-10.000xg), losses of linezolid were observed with the Nanosep Omega device due to adsorption onto the polyethersulfone membrane (unbound fraction 75% at 100mg/L and 45% at 0.1mg/L, respectively). No losses were observed with Vivacon which is equipped with a membrane of regenerated cellulose. With fluconazole no differences between Nanosep and Vivacon were observed. Applying standard conditions (pH 7.4/37°C/1000xg/20min), the mean unbound fraction of linezolid in pooled plasma from healthy volunteers was 81.5±2.8% using Vivacon, that of fluconazole was 87.9±3.5% using Nanosep or 89.4±3.3% using Vivacon. The unbound fraction of linezolid was 85.4±3.7% in plasma samples from surgical patients and 92.1±6.2% in ICU patients, respectively. The unbound fraction of fluconazole was 93.9±3.3% in plasma samples from ICU patients.

  4. Concentrations of haptoglobin in bovine plasma determined by ELISA or a colorimetric method based on peroxidase activity.

    Science.gov (United States)

    Cooke, R F; Arthington, J D

    2013-06-01

    The objective was to compare different procedures for determination of haptoglobin in bovine plasma. Nine Angus steers were vaccinated against Mannheimia haemolytica to stimulate an acute-phase response. Blood samples were collected immediately prior to vaccination (day 0), and on days 1, 3, 5, 7 and 10. Plasma samples were frozen in duplicates at -80 °C. One set of the duplicates was analysed for haptoglobin concentrations using a commercial ELISA kit. A day effect was detected (p peroxidase activity (CPPA) with results expressed as optical density. Further, based on the ELISA results, the plasma sample with the greatest haptoglobin concentration was also serially diluted into a plasma sample with negligible haptoglobin concentration from the same steer (1:1 through 1:1024 dilution). These dilutions were used within the CPPA method to generate a standard curve and estimate plasma haptoglobin concentrations (CPPA + STD). A linear standard curve was generated (r(2)  = 0.99). A day effect similar to the ELISA method was detected for the CPPA and CPPA + STD methods (p ELISA methods were positively correlated (r = 0.97; p ELISA method. In conclusion, assessing concentrations of haptoglobin in bovine plasma using the CPPA and CPPA + STD methods generate highly correlated or similar results, respectively, compared to ELISA. Therefore, the CPPA + STD and CPPA methods can be used as a less expensive alternative to ELISA to determine concentrations or monitor changes in plasma haptoglobin in bovine samples.

  5. Determining plasma morphine levels using GC-MS after solid phase extraction to monitor drug levels in the postoperative period

    Directory of Open Access Journals (Sweden)

    Veronica Santos

    2008-01-01

    Full Text Available OBJECTIVE: To implement a selective and sensitive analytical method to quantify morphine in small volumes of plasma by gas-liquid chromatography-mass spectrometry (GC-MS, aimed at post-operatively monitoring the drug. METHOD: A gas-liquid chromatographic method with mass detection has been developed to determine morphine concentration in plasma after solid phase extraction. Morphine-d3 was used as an internal standard. Only 0.5 mL of plasma is required for the drug solid-phase extraction in the Bond Elut-Certify®, followed by the quantification of morphine derivative by GC-MS using a linear temperature program, a capillary fused silica column, and helium as the carrier and make-up gas. The method was applied to determine morphine content in plasma samples of four patients during the postoperative period of cardiac surgery. Patient-controlled analgesia with morphine was performed by a venous catheter, and a series of venous blood samples were collected. After the oro-After the orotracheal extubation, morphine plasma levels were monitored for up to 36 hours. RESULTS: The run time was 16 minutes because morphine and the internal standard were eluted after 8.8 minutes. The GC-MS method had 0.5 -1000 ng/mL linearity range (r²=0.9995, 0.1 ng/mL limit of detection, intraday and interday precision equivalent to 1.9% and 6.8%, and 0.1% and 0.8% systematic error (intraday and interday, respectively. The analytical method showed optimal absolute (98% and relative (100.7% recoveries. Morphine dose requirements and plasma levels are discussed. CONCLUSION: The analytical gas-liquid chromatography-mass spectrometry method is selective and adequate for morphine measurements in plasma for applications in clinical studies.

  6. Determining plasma morphine levels using GC-MS after solid phase extraction to monitor drug levels in the postoperative period.

    Science.gov (United States)

    Santos, Veronica; López, Karin Jannet Vera; Santos, Luciana Moraes; Yonamine, Mauricio; Carmona, Maria José Carvalho; Santos, Silvia Regina Cavani Jorge

    2008-06-01

    To implement a selective and sensitive analytical method to quantify morphine in small volumes of plasma by gas-liquid chromatography-mass spectrometry (GC-MS), aimed at post-operatively monitoring the drug. A gas-liquid chromatographic method with mass detection has been developed to determine morphine concentration in plasma after solid phase extraction. Morphine-d3 was used as an internal standard. Only 0.5 mL of plasma is required for the drug solid-phase extraction in the Bond Elut-Certify, followed by the quantification of morphine derivative by GC-MS using a linear temperature program, a capillary fused silica column, and helium as the carrier and make-up gas. The method was applied to determine morphine content in plasma samples of four patients during the postoperative period of cardiac surgery. Patient-controlled analgesia with morphine was performed by a venous catheter, and a series of venous blood samples were collected. After the oro-After the orotracheal extubation, morphine plasma levels were monitored for up to 36 hours. The run time was 16 minutes because morphine and the internal standard were eluted after 8.8 minutes. The GC-MS method had 0.5 -1000 ng/mL linearity range (r(2)=0.9995), 0.1 ng/mL limit of detection, intraday and interday precision equivalent to 1.9% and 6.8%, and 0.1% and 0.8% systematic error (intraday and interday, respectively). The analytical method showed optimal absolute (98%) and relative (100.7%) recoveries. Morphine dose requirements and plasma levels are discussed. The analytical gas-liquid chromatography-mass spectrometry method is selective and adequate for morphine measurements in plasma for applications in clinical studies.

  7. High performance liquid chromatography for the determination of glucosamine sulfate in human plasma after derivatization with 9-fluorenylmethyl chloroformate.

    Science.gov (United States)

    Huang, Tao-min; Deng, Chun-Hui; Chen, Nian-zhu; Liu, Zhen; Duan, Geng-Li

    2006-10-01

    In this study, we developed a simple, rapid, sensitive, and reliable method for the determination of glucosamine sulfate in human plasma, which was based on derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by reverse-phase HPLC-FLD. For the first time, FMOC-Cl was introduced into derivatization of glucosamine sulfate in human plasma. The amino groups of glucosamine sulfate and vertilmicin sulfate (the internal standard) were trapped with FMOC-Cl to form glucosamine-FMOC-Cl and vertilmicin-FMOC-Cl adducts, which can be very suitable for HPLC-FLD. Precipitation of plasma proteins by acetonitrile was followed by vortex mixing and centrifugation. Chromatographic separation was performed on a C18 column (DIAMONSIL 150 x 4 mm id, 5 microm) with a mobile phase gradient consisting of acetonitrile and water at a flow-rate of 1 mL/min. The retention times of glucosamine-FMOC-Cl and vertilmicin-FMOC-Cl adducts were 8.9 and 21.2 min, respectively. This method was shown to be selective and sensitive for glucosamine sulfate. The limit of detection was 15 ng/mL for glucosamine sulfate in plasma and the linear range was 0.1-10 mg/mL in plasma with a correlation coefficient (r) of 0.9999. The relative standard deviations (RSDs) of intra-day and inter-day assays were 5.2-8.1% and 6.1- 8.5%, respectively. Extraction recoveries of glucosamine sulfate in plasma were greater than 90%. The validated method was successfully applied to the determination of glucosamine sulfate in human plasma samples.

  8. Bioanalytical method development and validation for determination of metoprolol tartarate and hydrochlorothiazide using HPTLC in human plasma

    Directory of Open Access Journals (Sweden)

    Ambadas Ranganath Rote

    2013-12-01

    Full Text Available A simple, sensitive, rapid and economic chromatographic method has been developed for determination of metoprolol tartarate and hydrochlorothiazide in human plasma using paracetamol as an internal standard. The analytical technique used for method development was high-performance thin-layer chromatography. HPTLC Camag with precoated silica gel Plate 60F254 (20 cm×10 cm at 250 µm thicknesses (E. Merck, Darmstadt, Germany was used as the stationary phase. The mobile phase used consisted of chloroform: methanol: ammonia (9:1:0.5v/v/v. Densitometric analysis was carried out at a wavelength of 239 nm. The rf values for hydrochlorothiazide, paracetamol and metoprolol tartarate were 0.13±0.04, 0.28±0.05, 0.48±0.04, respectively. Plasma samples were extracted by protein precipitation with methanol. Concentration ranges of 200, 400, 600, 800, 1000, 1200 ng/mL and 2000, 4000, 6000, 8000, 10000, 12000 ng/mL of hydrochlorothiazide and metoprolol tartarate, respectively, were used with plasma for the calibration curves. The percent recovery of metoprolol tartarate and hydrochlorothiazide was found to be 77.30 and 77.02 %, respectively. The stability of metoprolol tartarate and hydrochlorothiazide in plasma were confirmed during three freeze-thaw cycles (-20 ºC on a bench for 24 hours and post-preparatively for 48 hours. The proposed method was validated statistically and proved suitable for determination of metoprolol tartarate and hydrochlorothiazide in human plasma.

  9. A simple high-performance liquid chromatography for the determination of linezolid in human plasma and saliva.

    Science.gov (United States)

    Hara, Shuuji; Uchiyama, Masanobu; Yoshinari, Masami; Matsumoto, Taichi; Jimi, Shiro; Togawa, Atsushi; Takata, Tohru; Takamatsu, Yasushi

    2015-09-01

    Linezolid is an antimicrobial agent for the treatment of multiresistant Gram-positive infections. A practical high-performance liquid chromatography method was developed for the determination of linezolid in human plasma and saliva. Linezolid and an internal standard (o-ethoxybenzamide) were extracted from plasma and saliva with ethyl acetate and analyzed on a Capcell Pak C18 MG column with UV detection at 254 nm. The calibration curve was linear through the range 0.5-50 µg/mL using a 200 μL sample volume. The intra- and interday precisions were all saliva. The accuracies ranged from 98.8 to 110% for both matrices. The mean recoveries of linezolid were 80.8% for plasma and 79.0% for saliva. This method was used to determine the plasma and saliva concentrations of linezolid in healthy volunteers who were orally administered a 600 mg dose of linezolid. Our liquid-liquid extraction procedure is easy and requires a small volume of plasma or saliva (200 μL). This small volume can be advantageous in clinical pharmacokinetic studies, especially if children participate.

  10. Determination of Ephedrine Alkaloids in Human Urine and Plasma by Liquid Chromatography/Tandem Mass Spectrometry: Collaborative Study

    Science.gov (United States)

    Trujillo, William A.; Sorenson, Wendy R.

    2008-01-01

    A collaborative study was conducted to evaluate the accuracy and precision of a method for ephedrine-type alkaloids (i.e., norephedrine, norpseudoephedrine, ephedrine, pseudoephedrine, methylephedrine, and methylpseudoephedrine) in human urine and plasma. The amount of ephedrine-type alkaloids present was determined using liquid chromatography (LC) with tandem mass selective detection. The test samples were diluted to reflect a concentration of 5.00–100 ng/mL for each alkaloid. An internal standard was added and the alkaloids were separated using a 5 μm phenyl LC column with an ammonium acetate, glacial acetic acid, acetonitrile, and water mobile phase. Eight blind duplicates of human urine and eight blind duplicates of human plasma were analyzed by 10 collaborators. In addition to negative controls, test portions of urine and plasma were fortified at 3 different levels with each of the 6 ephedrine-type alkaloids at approximately 1, 2, and 5 μg/mL for urine and 100, 200, and 500 ng/mL for plasma. On the basis of the accuracy and precision results for this collaborative study, it is recommended that this method be adopted Official First Action for the determination of 6 different ephedrine-type alkaloids in human urine and plasma. PMID:14509420

  11. Progesterone and Mental Rotation Task: Is There Any Effect?

    Directory of Open Access Journals (Sweden)

    Donatas Noreika

    2014-01-01

    Full Text Available Mental rotation task (MRT incorporates elements of spatial abilities, important in many professions, with people of both genders involved. Importantly, these are the areas where spatial tasks might be performed for long time periods; thus adverse effects of mental fatigue are highly unwanted. Substantial variation of MRT performance in relation to estrogen levels has been observed in many studies, whereas the role of progesterone remains elusive. Here we aimed to elucidate the effect of progesterone level on the long-duration (1.5 hours performance of MRT. We included three groups of subjects: a group of males as a control, a group of females in their follicular phase (low progesterone and a group of females in their luteal phase (high progesterone, MRT accuracy and response time, subjective fatigue ratings and cardiovascular measures together with 17β-estradiol and progesterone concentrations were analyzed. We found that subjective ratings of fatigue increased, performance accuracy increased, and mean response times decreased during the task in all groups. Females in luteal phase were significantly slower not only than men, but also than females in their follicular phase. An increase in subjective fatigue ratings was positively related to progesterone level—at higher progesterone levels, females felt more tired.

  12. Determination of uranium from nuclear fuel in environmental samples using inductively coupled plasma mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Boulyga, S.F. [Forschungszentrum Juelich GmbH (Germany). Zentralabteilung fuer Chemische Analysen]|[Radiation Physics and Chemistry Problems Inst., Minsk (Belarus); Becker, J.S. [Forschungszentrum Juelich GmbH (Germany). Zentralabteilung fuer Chemische Analysen

    2000-11-01

    As a result of the accident at the Chernobyl nuclear power plant (NPP) the environment was contaminated with spent nuclear fuel. The {sup 236}U isotope was used in this study to monitor the spent uranium from nuclear fallout in soil samples collected in the vicinity of the Chernobyl NPP. A rapid and sensitive analytical procedure was developed for uranium isotopic ratio measurement in environmental samples based on inductively coupled plasma quadrupole mass spectrometry with a hexapole collision cell (HEX-ICP-QMS). The figures of merit of the HEX-ICP-QMS were studied with a plasma-shielded torch using different nebulizers (such as an ultrasonic nebulizer (USN) and Meinhard nebulizer) for solution introduction. A {sup 238}U{sup +} ion intensity of up to 27000 MHz/ppm in HEX-ICP-QMS with USN was observed by introducing helium into the hexapole collision cell as the collision gas at a flow rate of 10 ml min{sup -1}. The formation rate of uranium hydride ions UH{sup +}/U{sup +} of 2 x 10{sup -6} was obtained by using USN with a membrane desolvator. The limit of {sup 236}U/{sup 238}U ratio determination in 10 {mu}g 1{sup -1} uranium solution was 3 x 10{sup -7} corresponding to the detection limit for {sup 236}U of 3 pg 1{sup -1}. The precision of uranium isotopic ratio measurements in 10 {mu}g 1{sup -1} laboratory uranium isotopic standard solution was 0.13% ({sup 235}U/{sup 238}U) and 0.33% ({sup 236}U/{sup 238}U) using a Meinhard nebulizer and 0.45% ({sup 235}U/{sup 238}U) and 0.88% ({sup 236}U/{sup 238}U) using a USN. The isotopic composition of all investigated Chernobyl soil samples differed from those of natural uranium; i.e. in these samples the {sup 236}U/{sup 238}U ratio ranged from 10{sup -5} to 10{sup -3}. (orig.)

  13. Application of laser-produced-plasmas to determination of carbon content in steel; Aplicacion de los plasmas generados por laser a la determinacion de carbono en aceros

    Energy Technology Data Exchange (ETDEWEB)

    Ortiz, M.; Aragon, C.; Aguilera, J. A.; Campos, J.

    1994-07-01

    This paper describes an analytical method to determine carbon content in solid and molten steel. It is based on the study of the emission spectrum from a Nd-YAG laser produced plasma. The light emitted from the plasma is focused to the entrance slit of a spectrometer and detected by an OMA III system. For every laser pulse an spectral range of 100 A are recorded. With the use of time-resolved spectroscopy a precision of 1.6 % and a detection limit of 65 ppm of carbon content in steel have been obtained. These values are similar to those of other accurate conventional techniques but using optics fiber and laser excitation it is possible to made sample calibrations in hostile environments. Also, as the analysis are made in real time changes in sample composition can be measured without stopping production processes. (Author) 26 refs.

  14. Assessment of 7.5% NaCl /6% Dextran-70 (HSD) Effects on Serum or Plasma Protein Determinations

    Science.gov (United States)

    1990-12-26

    determined by modified Lowry, dye-binding, and an automated biuret method, as well as by refractometry , before, and at various times following HSD...protein concentrations determined by the biuret assay or refractometry when dextran serum concentrations exceeded 1.2 g/dl. The in vivo studies... refractometry , before, and at various times following HSD infusion in both euvolemic and hemorrhaged animals. Other studies analyzed plasma protein

  15. Progesterone promotes propagation and viability of mouse embryonic stem cells.

    Science.gov (United States)

    Shen, Shan-Wei; Song, Hou-Yan

    2009-10-25

    It has been known that estrogen-17beta stimulates proliferation of mouse embryonic stem (mES) cells. To explore the function of another steroid hormone progesterone, we used MTT method and BrdU incorporation assay to obtain growth curves, clone forming assay to detect the propagation and viability of individual mES cells, Western blot to test the expression of ES cell marker gene Oct-4, fluorescence activated cell sorter (FACS) to test cell cycle, and real-time PCR to detect the expressions of cyclins, cyclin-dependent kinases and proto-oncogenes. The results showed that progesterone promoted proliferation of mES cells. The number of clones was more in progesterone-treated group than that in the control group. The expression of pluripotency-associated transcriptional factor Oct-4 changed little after progesterone treatment as shown by Western blot, indicating that most of mES cells were in undifferentiated state. The results of FACS proved that progesterone promoted DNA synthesis in mES cells. The proportion of mES cells in S+G(2)/M phase was higher in progesterone-treated group than that in the control group. Cyclins and cyclin-dependent kinases, as well as proto-oncogenes (c-myc, c-fos) were up-regulated when cells were treated with progesterone. The results obtained indicate that progesterone promotes propagation and viability of mES cells. The up-regulation of cell cycle-related factors might contribute to the function of progesterone.

  16. Continuous Emission Spectrum Measurement for Electron Temperature Determination in Low-Temperature Collisional Plasmas%Continuous Emission Spectrum Measurement for Electron Temperature Determination in Low-Temperature Collisional Plasmas

    Institute of Scientific and Technical Information of China (English)

    刘秋艳; 李弘; 陈志鹏; 谢锦林; 刘万东

    2011-01-01

    Continuous emission spectrum measurement is applied for the inconvenient diagnos- tics of low-temperature collisional plasmas. According to the physical mechanism of continuous emission, a simplified model is presented to analyze the spectrum in low temperature plasma. The validity of this model is discussed in a wide range of discharge parameters, including electron tem- perature and ionization degree. Through the simplified model, the continuous emission spectrum in a collisional argon internal inductively coupled plasma is experimentally measured to determine the electron temperature distribution for different gas pressures and radio-frequency powers. The inverse Abel transform is also applied for a better spatially resoluted results. Meanwhile, the result of the continuous emission spectrum measurement is compared to that of the electrostatic double probes, which indicates the effectiveness of this method.

  17. Circulating levels of prolactin and progesterone in a wild population of red kangaroos (Macropus rufus) Marsupialia: Macropodidae

    Science.gov (United States)

    Muths, E.; Hinds, L. A.

    1996-01-01

    Circulating progesterone and prolactin levels were measured in shot and live-caught wild red kangaroos using radioimmunoassays validated for the red kangaroo. The objective of the study was to correlate hormone profiles with reproductive status and determine if red kangaroos follow the general pattern elucidated for other macropodids. During Phase 2a lactation (600 pg/ml (n= 32) during the transition to Phase 3 lactation (181 to 235 days) when the quiescent corpus luteum and embryo were reactivated. Progesterone concentrations then decreased to prolactin during Phase 2a were prolactin concentrations increased to 15 ng/ml (n= 32), then decreased and remained low through the subsequent stage of dual lactation. These results indicate that progesterone and prolactin profiles in wild red kangaroos follow patterns found previously in other macropodid species, the tammar and Bennett's wallabies.

  18. Mammary tumors and serum hormones in the bitch treated with medroxyprogesterone acetate or progesterone for four years

    Energy Technology Data Exchange (ETDEWEB)

    Frank, D.W.; Kirton, K.T.; Murchison, T.E.; Quinlan, W.J.; Coleman, M.E.; Gilbertson, T.J.; Feenstra, E.S.; Kimball, F.A.

    1978-01-01

    After four years of a long term contraceptive steroid safety study, the incidence and the histologic type of mammary dysplasia produced is similar in beagles treated with medroxyprogesterone acetate (medroxyprogesterone) or progesterone. Serum insulin, thyroid stimulating hormone (TSH), triiodothyronine, growth hormone, prolactin, 17..beta..-estradiol, progesterone, and cortisol were determined by radioimmunoassay on samples collected after 45 months of treatment. Serum growth hormone and insulin concentrations were elevated in a dose related manner in both treatment groups. Triiodothyronine, cortisol, and estradiol-17..beta.. (medroxyprogesterone only) were lowered. TSH and prolactin concentrations were not changed. Pituitary--gonadal hormone interaction in the pathogenesis of mammary neoplasia of the dog is discussed. Prolonged treatment of the beagle with massive doses of progesterone or medroxyprogesterone results in a dose related incidence of mammary modules.

  19. [Determination of 10 sedative-hypnotics in human plasma using pulse splitless injection technique and gas chromatography-mass spectrometry].

    Science.gov (United States)

    Chang, Qing; Ma, Hongying; Wang, Fangjie; Ou, Honglian; Zou, Ming

    2011-11-01

    A simple, precise and sensitive gas chromatography-mass spectrometry (GC-MS) method coupled with pulse splitless injection technique was developed for the determination of 10 sedative-hypnotics (barbital, amobarbital, phenobarbital, oxazepam, diazepam, nitrazepam, clonazepam, estazolam, alprazolam, triazolam) in human plasma. The drugs spiked in plasma were extracted with ethyl acetate after alkalization with 0.1 mol/L NaOH solution. The organic solvent was evaporated under nitrogen stream, and the residues were redissolved by ethyl acetate. The separation was performed on an HP-5MS column (30 m x 250 microm x 0.25 microm). The analytes were determined and identified using selected ion monitoring (SIM) mode and scan mode, respectively. The internal standard method was used for the determination. The target analytes were well separated from each other on their SIM chromatograms and also on the total ion current (TIC) chromatograms. The blank extract from human plasma gave no peaks that interfered with all the analytes on the chromatogram. The calibration curves for 10 sedative-hypnotics showed excellent linearity. The correlation coefficients of all the drugs were higher than 0.9954. The recoveries of the drugs spiked in human plasma ranged from 92.28% to 111.7%, and the relative standard deviations (RSDs) of intra-day and inter-day determinations were from 4.09% to 14.26%. The detection limits ranged from 2 to 20 microg/L. The method is simple, reliable, rapid and sensitive for the determination and the quantification of 10 sedative-hypnotics in human plasma and seems to be useful in the practice of clinical toxicological cases.

  20. Gamma-hydroxybutyrate enhances mood and prosocial behavior without affecting plasma oxytocin and testosterone.

    Science.gov (United States)

    Bosch, Oliver G; Eisenegger, Christoph; Gertsch, Jürg; von Rotz, Robin; Dornbierer, Dario; Gachet, M Salomé; Heinrichs, Markus; Wetter, Thomas C; Seifritz, Erich; Quednow, Boris B

    2015-12-01

    Gamma-hydroxybutyrate (GHB) is a GHB-/GABAB-receptor agonist. Reports from GHB abusers indicate euphoric, prosocial, and empathogenic effects of the drug. We measured the effects of GHB on mood, prosocial behavior, social and non-social cognition and assessed potential underlying neuroendocrine mechanisms. GHB (20mg/kg) was tested in 16 healthy males, using a randomized, placebo-controlled, cross-over design. Subjective effects on mood were assessed by visual-analogue-scales and the GHB-Specific-Questionnaire. Prosocial behavior was examined by the Charity Donation Task, the Social Value Orientation test, and the Reciprocity Task. Reaction time, memory, empathy, and theory-of-mind were also tested. Blood plasma levels of GHB, oxytocin, testosterone, progesterone, dehydroepiandrosterone (DHEA), cortisol, aldosterone, and adrenocorticotropic-hormone (ACTH) were determined. GHB showed stimulating and sedating effects, and elicited euphoria, disinhibition, and enhanced vitality. In participants with low prosociality, the drug increased donations and prosocial money distributions. In contrast, social cognitive abilities such as emotion recognition, empathy, and theory-of-mind, and basal cognitive functions were not affected. GHB increased plasma progesterone, while oxytocin and testosterone, cortisol, aldosterone, DHEA, and ACTH levels remained unaffected. GHB has mood-enhancing and prosocial effects without affecting social hormones such as oxytocin and testosterone. These data suggest a potential involvement of GHB-/GABAB-receptors and progesterone in mood and prosocial behavior. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Microwave diagnostic for the determination of the electron temperature of a low density shock-heated argon plasma

    Science.gov (United States)

    Schneider, C. P.; Exberger, R. J.

    1978-01-01

    The diffraction, defocusing and beam bending effects of microwaves transmitted through a bounded shock-heated argon plasma with low electron density are investigated with the purpose of developing an accurate method for electron temperature (Te) determination. This report describes the evaluation technique and presents results for an argon plasma with pressure between 3 and 10 torr, and T2 between 5500 and 9500 K. The electron temperature values obtained have a range of uncertainty between -20% and +10% only. The electron temperature is equal or lower by approximately 1000 K in comparison to the heavy particle temperature (T2).

  2. Roles of estrogen and progesterone in modulating renal nerve function in the rat kidney

    Energy Technology Data Exchange (ETDEWEB)

    Graceli, J.B. [Departamento de Morfologia, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES (Brazil); Cicilini, M.A.; Bissoli, N.S.; Abreu, G.R.; Moysés, M.R. [Departamento de Ciências Fisiológicas, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES (Brazil)

    2013-07-02

    The maintenance of extracellular Na{sup +} and Cl{sup -} concentrations in mammals depends, at least in part, on renal function. It has been shown that neural and endocrine mechanisms regulate extracellular fluid volume and transport of electrolytes along nephrons. Studies of sex hormones and renal nerves suggested that sex hormones modulate renal function, although this relationship is not well understood in the kidney. To better understand the role of these hormones on the effects that renal nerves have on Na{sup +} and Cl{sup -} reabsorption, we studied the effects of renal denervation and oophorectomy in female rats. Oophorectomized (OVX) rats received 17β-estradiol benzoate (OVE, 2.0 mg·kg{sup -1}·day{sup -1}, sc) and progesterone (OVP, 1.7 mg·kg{sup -1}·day{sup -1}, sc). We assessed Na{sup +} and Cl{sup -} fractional excretion (FE{sub Na{sup {sub +}}} and FE{sub Cl{sup {sub -}}}, respectively) and renal and plasma catecholamine release concentrations. FE{sub Na{sup {sub +}}}, FE{sub Cl{sup {sub -}}}, water intake, urinary flow, and renal and plasma catecholamine release levels increased in OVX vs control rats. These effects were reversed by 17β-estradiol benzoate but not by progesterone. Renal denervation did not alter FE{sub Na{sup {sub +}}}, FE{sub Cl{sup {sub -}}}, water intake, or urinary flow values vs controls. However, the renal catecholamine release level was decreased in the OVP (236.6±36.1 ng/g) and denervated rat groups (D: 102.1±15.7; ODE: 108.7±23.2; ODP: 101.1±22.1 ng/g). Furthermore, combining OVX + D (OD: 111.9±25.4) decreased renal catecholamine release levels compared to either treatment alone. OVE normalized and OVP reduced renal catecholamine release levels, and the effects on plasma catecholamine release levels were reversed by ODE and ODP replacement in OD. These data suggest that progesterone may influence catecholamine release levels by renal innervation and that there are complex interactions among renal nerves, estrogen

  3. Progesterone induces cellular differentiation in MDA-MB-231 breast cancer cells transfected with progesterone receptor complementary DNA.

    Science.gov (United States)

    Lin, Valerie Chun-Ling; Jin, Rongxian; Tan, Puay-Hoon; Aw, Swee-Eng; Woon, Chow-Thai; Bay, Boon-Huat

    2003-06-01

    Progesterone is an important regulator of growth and differentiation in breast tissues. In this study, the effect of progesterone on cell differentiation was evaluated in the estrogen receptor-negative and progesterone receptor (PR)-negative MDA-MB-231 cell line which was transfected with PR-complementary DNA. Morphological changes were analyzed at the ultrastructural level by scanning and transmission electron microscopy. Progesterone-treated PR-transfected cells exhibited a more protracted and well spread morphology with an increase in organelles such as mitochondria and rough endoplasmic reticulum as compared to the rounded form of control vehicle (0.1% ethanol)-treated PR-transfected cells. Vehicle and progesterone-treated MDA-MB-231 cells transfected with the pSG5 plasmid (transfection control cells) had similar rounded morphology as control vehicle-treated PR-transfected cells. Immunofluorescence staining revealed that expression of E-cadherin, a differentiation marker, was more prominent in progesterone-treated cells. Expression of keratin and vimentin but not beta-catenin was up-regulated in progesterone treated cells when evaluated by immunoblotting. As signal transducers and activators of transcription (STAT) molecules have been implicated in mammary differentiation, we analyzed the expression of Stat 1, 3, 5a, and 5b proteins and found a significant up-regulation of the Stat 5b protein in progesterone-treated cells. We have provided in vitro evidence of the close association of PR with differentiation in breast cancer. It is likely that the Stat 5b protein may play a major role in progesterone-induced differentiation in breast cancer cells.

  4. Determination of roxithromycin in human plasma by HPLC with fluorescence and UV absorbance detection: application to a pharmacokinetic study.

    Science.gov (United States)

    Główka, Franciszek K; Karaźniewicz-Łada, Marta

    2007-06-01

    A selective HPLC method with fluorescence detection for the determination of roxithromycin (ROX) in human plasma was described. After solid-phase extraction (SPE), ROX and erythromycin (internal standard, I.S.) were derivatized by treatment with 9-fluorenylmethyl chloroformate (FMOC-Cl). Optimal resolution of fluorescence derivatives of ROX and I.S. was obtained during one analytical run using reversed phase, C(18) column. The mobile phase was composed of potassium dihydrogenphosphate solution, pH 7.5 and acetonitrile. Fluorescence of the compounds was measured at the maximum excitation, 255 nm and emission, 313 nm, of ROX derivatives. Validation parameters of the method were also established. After SPE, differences in recoveries of ROX and erythromycin from human plasma were observed. The linear range of the standard curve of ROX in plasma was 0.5-10.0 mg/l. The validated method was successfully applied for pharmacokinetic studies of ROX after administration of a single tablet of ROX.

  5. Real-time DSP-based shape determination and plasma position control in the ISTTOK tokamak

    Energy Technology Data Exchange (ETDEWEB)

    Carvalho, B. E-mail: bernardo@cfn.ist.utl.pt; Fernandes, H.; Silva, C.; Borba, D.; Varandas, C.A.F

    2004-06-01

    We have developed a digital signal processor-based system for real-time control of the ISTTOK plasma position based on a low-cost digital signal processor (DSP) board. A novel plasma shape reproduction method was developed using a Cauchy-condition expansion applied to the vacuum region around the plasma. This method showed to be best suited than classic methods such as Legendre-Fourier expansion (LFE) or current filaments (CF), particularly in small tokamaks with passive stabilizer conductors where the presence of strong eddy currents can cause significant errors in magnetic field sensors. We compare the results of the boundary reconstruction method with the measurements from a microwave interferometer diagnostic. We present an implementation in the DSP system that allows the real time control of the plasma position with a required 1 ms period.

  6. Simultaneous Determination of Silybin A and Silybin B in Rat Plasma and Pharmacokinetic Study

    Institute of Scientific and Technical Information of China (English)

    CHU Yang; LI Wei; LI Zhi-wen; LI Xin-xin; MA Xiao-hui; ZHOU Shui-ping; ZHU Yong-hong

    2011-01-01

    Objective To investigate the bioavailability and pharmacokinetics of silybin A and silybin B in rats,respectively.Methods Following iv and ig administration of silybin to 20 Wistar rats,the plasma samples were collected at different time points up to 12 h.Sample pretreatment was involved in one-step protein precipitation with acetonitrile.Silybin A and silybin B were simultaneously determined by LC-MS/MS.Results After ig dosing silybin 28,56,and 112 mg/kg to rats,the t1/2β values were 5.48,5.08,and 5.73 h for silybin A,and 4.56,4.12,and 5.53 h for silybin B; The Cmax were 674.3,1349.4,and 2042.5 ng/mL for silybin A,and 671.0,1365.4,and 2066.2 ng/mL for silybin B; The Tmax were 0.20,0.23,and 0.20 h for silybin A,and 0.20,0.23,and 0.20 h for silybin B; The AUC were 454.4,845.9,and 1219.5 h·ng/mL for silybin A,and 432.0,817.1,and 1153.6 h·ng/mL for silybin B.The absolute bioavailabilities of silybin A and silybin B were 2.86% and 1.93%,respectively.Conclusion Silybin A and silybin B have very low bioavailability after ig administration,and there is no significant difference in the pharmacokinetic parameters between silybin A and silybin B,which indicates that the two diastereoisomers have similar pharmacokinetic behavior in rats.

  7. Determination of minor elements in steelmaking flue dusts using laser ablation inductively coupled plasma mass spectrometry.

    Science.gov (United States)

    Coedo, A G; Padilla, I; Dorado, M T

    2005-07-15

    Element determination in solid waste products from the steel industry usually involves the time-consuming step of preparing a solution of the solid. Laser ablation (LA) inductively coupled plasma mass spectrometry (ICP-MS) has been applied to the analysis of Cr, Ni, Cu, As, Cd and Sn, elements of importance from the point of view of their impact on the environment, in electric arc furnace flue dust (EAFD). A simple method of sample preparation as pressed pellets using a mixture of cellulose and paraffin as binder material was applied. Calibration standards were prepared spiking multielement solution standards to a 1:1 ZnO+Fe(2)O(3) synthetic matrix. The wet powder was dried and mechanically homogenised. Quantitative analysis were based on external calibration using a set of matrix matched calibration standards with Rh as a internal standard. Results obtained using only one-point for calibration without matrix matched, needing less time for standardization and data processing, are also presented. Data are calculated for flue dust reference materials: CRM 876-1 (EAFD), AG-6203 (EAFD), AG-6201 (cupola dust) and AG-SX3705 (coke ashes), and for two representative electrical arc furnace flue dusts samples from Spanish steelmaking companies: MS-1 and MS-2. For the reference materials, an acceptable agreement with certificate values was achieved, and the results for the MS samples matched with those obtained from conventional nebulization solutions (CN). The analytical precision was found to be better than 7% R.S.D. both within a single pellet and between several pellets of the same sample for all the elements.

  8. Determination of plasma cholesterol sulfate by LC-APCI-MS/MS in the context of pediatric autism.

    Science.gov (United States)

    Fong, Bonnie Mei-Wah; Tam, Sidney; Leung, Kelvin Sze-Yin

    2013-11-15

    Cholesterol sulfate (CS) has various biological functions. Previously, plasma CS was measured primarily as a means to diagnose X-linked ichthyosis; however, a recent hypothesis suggests that CS deficiency might be related to autism. As such, an assay capable of measuring both very high (in the case of X-linked ichthyosis) and very low (in the case of autism) plasma CS levels is required. Here we describe a novel LC-APCI-MS/MS method for the determination of CS in human plasma, and we propose normal CS ranges for children, based on studies of a local population of normal Chinese children between the ages of 2 and 10. In addition, we have used this method to measure plasma CS in autistic children. CS was isolated by solid-phase extraction, and quantified by isotope-dilution LC-APCI-MS/MS in negative ion mode monitoring 465.3>97.1 m/z (CS) and 472.3>97.1 m/z (CS-d7). Mean recovery of the assay ranged from 88.1 to 112.7%; within- and between-run imprecisions have CVs less than 7.2 and 8.1%, respectively. The assay was linear up to at least 100 µmol L(-1). The reference interval of plasma CS in males (range: 1.16-4.23 µmol L(-1)) was found to be higher than in females (range: 0.86-3.20 µmol L(-1)). Comparison of normal and autistic children showed no statistically significant difference in the plasma CS level. In conclusion, a robust LC-APCI-MS/MS method for plasma CS was developed, and a pediatric reference interval was derived from applying the method to normal and autistic children.

  9. High Performace Liquid Chromtographic Determination of Nicardipine Hydrochloride in Human Plasma

    Directory of Open Access Journals (Sweden)

    Y. S. R. Krishnaiah

    2004-01-01

    Full Text Available A sensitive high-performance liquid chromatographic method was developed for the estimation of nicardipine hydrochloride in human plasma. Varying amount of nicardipine hydrochloride (2.5 to 150 ng/0.5 mL and fixed quantity (100 ng/0.5 mL of nifedipine (internal standard was added to blank human plasma, and a single step extraction was carried out with ethyl acetate. The mixture was centrifuged, ethyl acetate layer separated, dried and reconstituted with 100 μL of acetonitrile. Twenty microliters of this solution was injected into a reverse phase C-18 column using a mobile phase consisting of acetonitrile: 0.02 M potassium dihydrogen phosphate (pH 4.0 in the ratio of 60:40 v/v and the eluents were monitored at 239 nm. The method was validated for its linearity, precision and accuracy. The calibration curve was linear in the range of 5-150 ng/0.5 mL of plasma and the lower detection limit was 2.5 ng/0.5 mL of plasma. The intra- and inter-day variation was found to be less than 2.5% indicating that the method is highly precise. The mean recovery of nicardipine hydrochloride from plasma samples was 89.6±2.60%. The proposed HPLC method was applied for the estimation of nicardipine hydrochloride in human plasma after oral administration of an immediate release nicardipine hydrochloride capsule (dose 30 mg to 6 adult male volunteers. There was no interference of either the drug metabolites or other plasma components with the proposed HPLC method for the estimation of nicardipine hydrochloride in human plasma. Due to its simplicity, sensitivity, high precision and accuracy, the proposed HPLC method may be used for biopharmaceutical and pharmacokinetic evaluation of nicardipine hydrochloride and its formulations in humans

  10. Determination of loperamide in human plasma and saliva by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Arafat, Tawfiq; Arafat, Basil; awad, Riad; awwad, Ahmad Abu

    2014-12-01

    A simple and sensitive liquid chromatography-tandem mass spectrometric method for quantification of loperamide in human plasma and saliva was developed and validated, and then successfully applied in pharmacokinetic clinical study to investigate and correlate bioavailability of Imodium(®) 2mg quartet tablet dose in both human plasma and saliva. Loperamide with labeled internal standard was extracted from its biological matrix by methanol as protein direct precipitant in single extraction step. Adequate chromatographic separation for analytes from plasma and saliva matrices was achieved using ACE C18 (50mm×2.1mm, 5μm) column, eluted by water/methanol/formic acid (30:70:0.1%, v/v), delivered isocratically at constant flow rate of 0.75ml/min. The method validation intends to investigate specificity, sensitivity, linearity, precision, accuracy, recovery, matrix effect and stability according to European guideline, and partial validation was applied on saliva, specificity, matrix effect, recovery, sensitivity, within and between day precision and accuracy. The calibration curve was linear through the range of 20-3000pg/ml in both plasma and saliva using a 50μl sample volume. The partial validation sections outcome in saliva was so close to those in plasma. The within- and between-day precisions were all below 8.7% for plasma and below 11.4% for saliva. Accuracies ranged from 94 to 105% for both matrices. In this study, 26 healthy volunteers participated in the clinical study, and 6 of gave their saliva samples in addition to plasma at the same time schedule. The pharmacokinetic parameters of Cmax, AUC0-t and AUC0-∞, Tmax and T1/2 in both plasma and saliva were calculated and correlated.

  11. Plasma-based determination of inorganic contaminants in waste of electric and electronic equipment after microwave-induced combustion

    Science.gov (United States)

    Mello, Paola A.; Diehl, Lisarb O.; Oliveira, Jussiane S. S.; Muller, Edson I.; Mesko, Marcia F.; Flores, Erico M. M.

    2015-03-01

    A systematic study was performed for the determination of inorganic contaminants in polymeric waste from electrical and electronic equipment (EEE) for achieving an efficient digestion to minimize interferences in determination using plasma-based techniques. The determination of As, Br, Cd, Co, Cr, Cu, Ni, Pb, Sb, and Zn by inductively coupled plasma mass spectrometry (ICP-MS) and also by inductively coupled plasma optical emission spectrometry (ICP OES) was carried out after digestion using microwave-induced combustion (MIC). Arsenic and Hg were determined by flow-injection chemical vapor generation inductively coupled plasma mass spectrometry (FI-CVG-ICP-MS). Dynamic reaction cell inductively coupled plasma mass spectrometry (DRC-ICP-MS) with ammonia was also used for Cr determination. The suitability of MIC for digestion of sample masses up to 400 mg was demonstrated using microcrystalline cellulose as aid for combustion of polymers from waste of EEEs that usually contain flame retardants that impair the combustion. The composition and concentration of acid solutions (HNO3 or HNO3 plus HCl) were evaluated for metals and metalloids and NH4OH solutions were investigated for Br absorption. Accuracy was evaluated by comparison of results with those obtained using high pressure microwave-assisted wet digestion (HP-MAWD) and also by the analysis of certified reference material (CRM) of polymer (EC680k-low-density polyethylene). Bromine determination was only feasible using digestion by MIC once losses were observed when HP-MAWD was used. Lower limits of detection were obtained for all analytes using MIC (from 0.005 μg g- 1 for Co by ICP-MS up to 3.120 μg g-1 for Sb by ICP OES) in comparison to HP-MAWD due to the higher sample mass that can be digested (400 mg) and the use of diluted absorbing solutions. The combination of HNO3 and HCl for digestion showed to be crucial for quantitative recovery of some elements, as Cr and Sb. In addition, suitable agreement of Cr to

  12. Bacteria-induced or bacterial product-induced preterm parturition in mice and rabbits is preceded by a significant fall in serum progesterone concentrations.

    Science.gov (United States)

    Fidel, P I; Romero, R; Maymon, E; Hertelendy, F

    1998-01-01

    Bacterial products are thought to induce labor by stimulating the production of pro-inflammatory cytokines and prostaglandins in gestational tissues, leading to the onset of preterm parturition. Progesterone withdrawal is a prerequisite of parturition in many species. Yet a role for progesterone in the mechanisms responsible for preterm parturition, in the setting of infection, is unclear. The current studies were conducted to determine if a fall in serum progesterone concentrations occurs before the onset of bacterial product-induced preterm parturition in animals. Accordingly, pregnant mice at day 15 (70% gestation) were injected i.p. with Escherichia coli lipopolysaccharide (LPS; 50 microg/mouse) and timed-pregnant rabbits were inoculated transcervically with a suspension of E. coli to cause an ascending intrauterine infection. Control animals in both groups received equal volumes of sterile phosphate-buffered saline (PBS) solution. Blood specimens were collected at regular intervals and serum progesterone levels were determined by RIA. Within 14 h of LPS administration, mice delivered their pups. The median concentrations of serum progesterone were significantly lower at 1 h, 4 h, 10 h, and at the onset of preterm parturition (11-12 h) after LPS injection, compared to that in animals given PBS. Similarly, E. coli-inoculated rabbits delivered 1-2 days posttranscervical inoculation and demonstrated 60% decrease in serum progesterone within 12-24 h of inoculation compared to those given PBS. Parturition in both control groups occurred at term, following typical progesterone withdrawal. It is concluded that LPS administration to pregnant mice and ascending intrauterine infection in pregnant rabbits is associated with a dramatic fall in serum progesterone concentrations prior to the onset of parturition.

  13. Relationships between Milk Progesterone Profiles and Genetic Merit for Milk Production, Milking Frequency, and Feeding Regimen in Dairy Cattle

    NARCIS (Netherlands)

    Windig, J.J.; Beerda, B.; Veerkamp, R.F.

    2008-01-01

    Milk progesterone profiles were determined from samples obtained twice weekly for 100 d postpartum in 100 Holstein primiparous cows at a Dutch experimental farm. Three treatments were applied in a 2 x 2 x 2 factorial arrangement with high-low genetic merit for overall production, high-low caloric

  14. Analysis of estrogen and progesterone receptors on preoperative fine-needle aspirates.

    Science.gov (United States)

    Frigo, B; Pilotti, S; Zurrida, S; Ermellino, L; Manzari, A; Rilke, F

    1995-01-01

    For 56 cases of carcinoma of the breast, results of the immunocytochemical assay for estrogen and progesterone receptors performed on preoperative fine-needle aspirates were compared with those obtained on scraping material from the same tumors. The value and usefulness of this last analysis was demonstrated in a previous study. The level of agreement between the two cytological techniques was assessed by the k statistic. A high level of agreement was found, with k values of 0.909 and 0.889 for estrogen and progesterone receptors, respectively. The results reported here revealed the reliability of steroid receptor determination on fine-needle aspiration biopsies, provided that sufficient cellularity was available. This technique can replace the open biopsy procedure, in as much as it represents a rapid, almost painless, and easily repeated method for the assessment of the receptor status, and is useful for treatment decisions at any time during the course of the disease.

  15. Determination of plasma trace elements in tumor-bearing animals by proton-induced X-ray emission spectroscopy.

    Science.gov (United States)

    Fogle, M; Daly, B; Evans, M; Justiniano, E L; Kovacs, C J; Shinpaugh, J L; Toburen, L H

    2001-11-01

    Although altered levels of circulating essential trace elements are known to accompany malignant disease, the lack of sensitivity of conventional detection methods has generally limited their study to clinical conditions involving extensive disease (i.e., significant tumor burden). As such, the application of altered trace element levels as potential prognostic guides or as response indicators subsequent to treatment has been of limited use. During this study, proton-induced X-ray emission spectroscopy was evaluated as a tool to determine trace element imbalances in a murine tumor model. Using plasma from C57B1/6 mice bearing the syngeneic Lewis lung carcinoma (LLCa), levels of Fe, Cu, and Zn, as well as changes in the Cu /Zn ratio, were measured in animals carrying an increasing primary tumor burden. The plasma levels of Fe, Cu, and Zn were found to decrease significantly 7 d following implants of LLCa cells with no significant change observed in the Cu/Zn ratio. By d 21, however, an increase in the Cu/Zn ratio was found to accompany increased growth of the LLCa tumor; the plasma levels of Cu had returned to normal levels, whereas both the Fe and Zn plasma levels remained lowered. Collectively, the results suggest that although a net change in individual plasma trace element concentrations might not be accurately associated with tumor growth, a clear relationship was established between the Cu/Zn ratio and tumor size.

  16. A hepatitis C avidity test for determining recent and past infections in both plasma and dried blood spots.

    Science.gov (United States)

    Shepherd, Samantha J; Kean, Joy; Hutchinson, Sharon J; Cameron, Sheila O; Goldberg, David J; Carman, William F; Gunson, Rory N; Aitken, Celia

    2013-05-01

    DBS testing has been used successfully to detect HCV antibody positive individuals. Determining how long someone has been infected is important for surveillance initiatives. Antibody avidity is a method that can be used to calculate recency of infection. A HCV avidity assay was evaluated for both plasma and DBS. To measure antibody avidity a commercial HCV ELISA was modified using 7 M urea. The plasma samples were split into: group 1 (recently infected N = 19), group 2 (chronic carrier N = 300) and group 3 (resolved infection N = 82). Mock DBS made from group 1 (N = 12), group 2 (N = 50), group 3 (N = 25) and two seroconverter panels were evaluated. 133 DBS taken from patients known to have a resolved infection or be a chronic carrier were also tested. The avidity assay cut-off was set at AI≤30 for a recent infection. Using sequential samples the assay could detect a recent infection in the first 4-5 months from the point of infection. Most of the false positive results (AI plasma and DBS; as a result, a testing algorithm has been designed incorporating both PCR and two dilution factors. The sensitivity and specificity of the assay on plasma was 100% and 99.3%, respectively, while DBS had 100% sensitivity and 98.3% specificity. The HCV avidity assay can be used to distinguish between chronic and recent infection using either plasma or DBS as the sample type. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Use of Dried Plasma Spots for HIV-1 Viral Load Determination and Drug Resistance Genotyping in Mexican Patients

    Science.gov (United States)

    Rodriguez-Auad, Juan Pablo; Rojas-Montes, Othon; Maldonado-Rodriguez, Angelica; Alvarez-Muñoz, Ma. Teresa; Muñoz, Onofre; Torres-Ibarra, Rocio; Vazquez-Rosales, Guillermo

    2015-01-01

    Monitoring antiretroviral therapy using measurements of viral load (VL) and the genotyping of resistance mutations is not routinely performed in low- to middle-income countries because of the high costs of the commercial assays that are used. The analysis of dried plasma spot (DPS) samples on filter paper may represent an alternative for resource-limited settings. Therefore, we evaluated the usefulness of analyzing DPS samples to determine VL and identify drug resistance mutations (DRM) in a group of HIV-1 patients. The VL was measured from 22 paired plasma and DPS samples. In these samples, the average VL was 4.7 log10 copies/mL in liquid plasma and 4.1 log10 copies/mL in DPS, with a correlation coefficient of R = 0.83. A 1.1 kb fragment of HIV pol could be amplified in 14/22 (63.6%) of the DPS samples and the same value was amplified in plasma samples. A collection of ten paired DPS and liquid plasma samples was evaluated for the presence of DRM; an excellent correlation was found in the identification of DRM between the paired samples. All HIV-1 pol sequences that were obtained corresponded to HIV subtype B. The analysis of DPS samples offers an attractive alternative for monitoring ARV therapy in resource-limited settings. PMID:26779533

  18. Use of Dried Plasma Spots for HIV-1 Viral Load Determination and Drug Resistance Genotyping in Mexican Patients

    Directory of Open Access Journals (Sweden)

    Juan Pablo Rodriguez-Auad

    2015-01-01

    Full Text Available Monitoring antiretroviral therapy using measurements of viral load (VL and the genotyping of resistance mutations is not routinely performed in low- to middle-income countries because of the high costs of the commercial assays that are used. The analysis of dried plasma spot (DPS samples on filter paper may represent an alternative for resource-limited settings. Therefore, we evaluated the usefulness of analyzing DPS samples to determine VL and identify drug resistance mutations (DRM in a group of HIV-1 patients. The VL was measured from 22 paired plasma and DPS samples. In these samples, the average VL was 4.7 log10 copies/mL in liquid plasma and 4.1 log10 copies/mL in DPS, with a correlation coefficient of R = 0.83. A 1.1 kb fragment of HIV pol could be amplified in 14/22 (63.6% of the DPS samples and the same value was amplified in plasma samples. A collection of ten paired DPS and liquid plasma samples was evaluated for the presence of DRM; an excellent correlation was found in the identification of DRM between the paired samples. All HIV-1 pol sequences that were obtained corresponded to HIV subtype B. The analysis of DPS samples offers an attractive alternative for monitoring ARV therapy in resource-limited settings.

  19. Determination of AJ-3941, a possible agent for the treatment of cerebrovascular disorders, in plasma and brain by means of high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Kurono, M; Yoshida, K; Naruto, S

    1992-07-01

    A sensitive and selective high-performance liquid chromatographic method with fluorescence detection is described for the determination of AJ-3941 (I), a possible agent for the treatment of cerebrovascular disorders, in plasma and brain tissue. A simple hexane extraction was used for plasma, and for brain homogenate the hexane extract was further purified by solid-phase extraction. The determination limit was ca. 3 ng/ml for both plasma (0.5 ml) and 10% (w/v) brain homogenate (1 ml). The method was applied to the determination of I in plasma and brain samples of experimental animals.

  20. Effect of subluteal concentrations of progesterone on luteinizing hormone and ovulation in lactating dairy cows.

    Science.gov (United States)

    Hatler, T B; Hayes, S H; Ray, D L; Reames, P S; Silvia, W J

    2008-09-01

    Two experiments were conducted to determine if administration of progesterone within a low, subluteal range (0.1-1.0 ng/mL) blocks the luteinizing hormone (LH) surge (experiments 1 and 2) and ovulation (experiment 2) in lactating dairy cows. In experiment 1, progesterone was administered to cycling, lactating dairy cows during the luteal phase of the estrous cycle using a controlled internal drug release (CIDR) device. CIDRs were pre-incubated in other cows for either 0 (CIDR-0), 14 (CIDR-14) or 28 days (CIDR-28). One group of cows received no CIDRs and served as controls. One day after CIDR insertion, luteolysis was induced by two injections of prostaglandin (PG) F(2alpha) (25 mg) at 12 h intervals. Two days after the first injection, estradiol cypionate (ECP; 3 mg) was injected to induce a LH surge. Concentrations of progesterone after luteolysis were 0.11, 0.45, 0.78 and 1.20 ng/mL for cows treated with no CIDR, CIDR-28, CIDR-14, and CIDR-0, respectively. LH surges were detected in 4/4 controls, 4/5 CIDR-28, 2/5 CIDR-14 and 0/5 CIDR-0 cows following ECP. In experiment 2, progesterone was administered to cycling, lactating, Holstein cows during the luteal phase of the estrous cycle as in experiment 1. Luteolysis was induced as in experiment 1. The occurrence of an endogenous LH surge and ovulation were monitored for 7 days. Concentrations of progesterone after luteolysis were 0.13, 0.30, 0.70 and 1.20 ng/mL for cows treated with no CIDR, CIDR-28, CIDR-14 and CIDR-0, respectively. LH surges and ovulation were detected in 5/5 controls, 3/7 CIDR-28, 0/5 CIDR-14 and 0/5 CIDR-0 cows. It was concluded that low concentrations of progesterone can reduce the ability of either endogenous or exogenous estradiol to induce a preovulatory surge of LH and ovulation.

  1. Effects of supplemental progesterone after artificial insemination on expression of interferon-stimulated genes and fertility in dairy cows.

    Science.gov (United States)

    Monteiro, P L J; Ribeiro, E S; Maciel, R P; Dias, A L G; Solé, E; Lima, F S; Bisinotto, R S; Thatcher, W W; Sartori, R; Santos, J E P

    2014-01-01

    The objectives of the current study were to evaluate the effects of supplemental progesterone after artificial insemination (AI) on expression of IFN-stimulated genes (ISG) in blood leukocytes and fertility in lactating dairy cows. Weekly cohorts of Holstein cows were blocked by parity (575 primiparous and 923 multiparous) and method of insemination (timed AI or AI on estrus) and allocated randomly within each block to untreated controls, a controlled internal drug release (CIDR) containing 1.38g of progesterone from d 4 to 18 after AI (CIDR4), or a CIDR on d 4 and another on d 7 after AI and both removed on d 18 (CIDR4+7). Blood was sampled to quantify progesterone concentrations in plasma and mRNA expression in leukocytes for the ubiquitin-like IFN-stimulated gene 15-kDa protein (ISG15) and receptor transporter protein-4 (RTP4) genes. Pregnancy was diagnosed on d 34±3 and 62±3 after AI. Treatment increased progesterone concentrations between d 5 and 18 after AI in a dose-dependent manner (control=3.42, CIDR4=4.97, and CIDR4+7=5.46ng/mL). Cows supplemented with progesterone tended to have increased luteolysis by d 19 after AI (control=17.2; CIDR4=29.1; CIDR4+7=30.2%), which resulted in a shorter AI interval for those reinseminated after study d 18. Pregnancy upregulated expression of ISG in leukocytes on d 19 of gestation, but supplementing progesterone did not increase mRNA abundance for ISG15 and RTP4 on d 16 after insemination and tended to reduce mRNA expression on d 19 after AI. For RTP4 on d 19, the negative effect of supplemental progesterone was observed only in the nonpregnant cows. No overall effect of treatment was observed on pregnancy per AI on d 62 after insemination and averaged 28.6, 32.7, and 29.5% for control, CIDR4, and CIDR4+7, respectively. Interestingly, an interaction between level of supplemental progesterone and method of AI was observed for pregnancy per AI. For cows receiving exogenous progesterone, the lower supplementation with CIDR4

  2. Progesterone is neuroprotective by inhibiting cerebral edema after ischemia

    Institute of Scientific and Technical Information of China (English)

    Yuan-zheng Zhao; Min Zhang; Heng-fang Liu; Jian-ping Wang

    2015-01-01

    Ischemic edema can alter the structure and permeability of the blood-brain barrier. Recent stud-ies have reported that progesterone reduces cerebral edema after cerebral ischemia. However, the underlying mechanism of this effect has not yet been elucidated. In the present study, pro-gesterone effectively reduced Evans blue extravasation in the ischemic penumbra, but not in the ischemic core, 48 hours after cerebral ischemia in rats. Progesterone also inhibited the down-reg-ulation of gene and protein levels of occludin and zonula occludens-1 in the penumbra. These results indicate that progesterone may effectively inhibit the down-regulation of tight junctions, thereby maintaining the integrity of the blood-brain barrier and reducing cerebral edema.

  3. Effect of Progesterone Therapy versus Diet Modification on ...

    African Journals Online (AJOL)

    Background: Pregnant women may experience constipation for the first time or ... Aim: To compare the effect of progesterone versus diet modification in the treatment of constipation .... is a natural product that contains several types of fiber.

  4. Corpus luteum development and function after supplementation of long-acting progesterone during the early luteal phase in beef cattle.

    Science.gov (United States)

    Pugliesi, G; Oliveria, M L; Scolari, S C; Lopes, E; Pinaffi, F V; Miagawa, B T; Paiva, Y N; Maio, J R G; Nogueira, G P; Binelli, M

    2014-02-01

    Strategic supplementation of P4 may be used to increase conception rates in cattle, but timing of supplementation in relation to ovulation, mass of supplementary P4 and formulation of the P4-containing supplement has not been determined for beef cattle. Effects of supplementation of long-acting progesterone (P4) on Days 2 or 3 post-ovulation on development, function and regression of corpus luteum (CL) were studied in beef cattle. Cows were synchronized with an oestradiol/P4-based protocol and treated with 150 or 300 mg of long-acting P4 on Day 2 or 3 post-ovulation (6-7 cows/group). Colour-doppler ultrasound scanning and blood sample collection were performed from Day 2-21.5. Plasma P4 concentrations were greater (p  0.05) among groups, suggesting no effect of P4 treatment on luteal development. The frequency of cows that began luteolysis before Day 15 was greater (p beef cattle. © 2013 Blackwell Verlag GmbH.

  5. A Comparison between Ion chromatography and Inductively Coupled Plasma for the Determination of Bromate in Certain Samples of Foodstuffs

    Directory of Open Access Journals (Sweden)

    Alanowd O. Mehder

    2015-06-01

    Full Text Available Ion chromatography (IC and inductively coupled plasma (ICP-MS both were applied for the determination of bromate in some food samples. Attempts were made to establish calibration curves, however in case of IC, an additional abnormal peak was found to overlap with the bromate peak. This renders IC to be unsuccessful in the determination of bromate compared to ICP-MS technique. ICP-MS was found to give accurate results; therefore, it was applied for the determination of bromate in different samples of food stuffs.

  6. Enhanced Human Decidual Cell-Expressed FKBP51 May Promote Labor-Related Functional Progesterone Withdrawal.

    Science.gov (United States)

    Schatz, Frederick; Guzeloglu-Kayisli, Ozlem; Basar, Murat; Buchwalder, Lynn F; Ocak, Nehir; Guzel, Elif; Guller, Seth; Semerci, Nihan; Kayisli, Umit A; Lockwood, Charles J

    2015-09-01

    Sustained plasma progesterone (P4) levels suggest initiation of human term labor by functional P4 withdrawal, reflecting reduced progesterone receptor (PR) and/or glucocorticoid receptor (GR) expression or activity. The steroid-induced immunophilin cochaperone FKBP51 inhibits PR- and GR-mediated transcription, suggesting a labor-initiating role. Gestational age-matched decidual sections were immunostained for FKBP51 and decidual cell (DC) and interstitial trophoblast (IT) markers, vimentin and cytokeratin, respectively. Term DC cultures were incubated with vehicle (control), estradiol (E2) with or without medroxyprogesterone acetate, dexamethasone (Dex), or Organon 2058. FKBP51 histologic scoring was significantly higher in DC nuclei during labor versus prelabor decidua, whereas FKBP51 immunostaining was undetected in interstitial trophoblasts (P Organon 2058 inhibited PR expression (P < 0.05), and E2 + Dex inhibited GR expression (P < 0.05). Unlike term DCs, FKBP51 was undetected in control or Dex-treated cultured third-trimester trophoblasts. Electrophoretic mobility shift assays revealed that FKPB51 overexpression or silencing in cultured DCs altered PR-DNA binding. Increased FKBP51 levels in term DCs during labor complement our prior in situ observations of significantly lower PR in labor versus prelabor DCs. In a milieu of sustained plasma P4 levels, these reciprocal changes will amplify functional P4 withdrawal in DCs via FKBP51-mediated PR resistance coupled with declining PR levels, whereas the lack of FKBP51 expression in interstitial trophoblasts suggests unopposed constitutive GR action.

  7. Simultaneous determination of triple therapy for Helicobacter pylori in human plasma by reversed phase chromatography with online wavelength switching

    Science.gov (United States)

    Ahmed, Sameh; Atia, Noha N.

    2015-02-01

    The infection of gastric mucosa by Helicobacter pylori (HP) is an essential cofactor in the aetiology of gastroduodenal ulcer and gastric carcinoma. Because of the bacterial resistance, combination therapy containing omeprazole (OME), tinidazole (TNZ) and clarithromycin (CLA) is commonly used for eradication of HP. However, the simultaneous determination of the triple therapy in human plasma was not reported. A simple, reproducible, and selective HPLC method was developed for the simultaneous determination of the triple therapy mixture used for management of HP infections in human plasma. An HPLC procedure based on a liquid-liquid extraction, enrichment of the analytes and subsequent reversed-phase chromatography with UV detection was used. To enable sensitive and selective detection, the method involved the use of online wavelength switching detection, with two different detection wavelengths; 280 nm for detection of OME and TNZ and 210 nm for detection of CLA. Separations were performed on C18 analytical column with acetonitrile-10 mM phosphate buffer of pH = 3.0 at flow rate of 1.0 mL min-1. The linear ranges in human plasma were 0.05-10 μg mL-1 with correlation coefficients >0.9990. The detection limits in human plasma were 0.02-0.07 μg mL-1. Validation parameters were assessed in compliance with US-FDA guidelines. The method proved to be valuable for the therapeutic drug monitoring after oral administration of triple therapy tablets.

  8. Quantitative determination of fentanyl in newborn pig plasma and cerebrospinal fluid samples by HPLC-MS/MS.

    Science.gov (United States)

    Blanco, M E; Encinas, E; González, O; Rico, E; Vozmediano, V; Suárez, E; Alonso, R M

    2015-09-01

    In this study, a selective and sensitive high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method requiring low sample volume (≤100 μL) was developed and validated for the quantitative determination of the opioid drug fentanyl in plasma and cerebrospinal fluid (CSF). A protein precipitation extraction with acetonitrile was used for plasma samples whereas CSF samples were injected directly on the HPLC column. Fentanyl and (13) C6 -fentanyl (Internal Standard) were analyzed in an electrospray ionization source in positive mode, with multiple reaction monitoring (MRM) of the transitions m/z 337.0/188.0 and m/z 337.0/105.0 for quantification and confirmation of fentanyl, and m/z 343.0/188.0 for (13) C6 -fentanyl. The respective lowest limits of quantification for plasma and CSF were 0.2 and 0.25 ng/mL. Intra- and inter-assay precision and accuracy did not exceed 15%, in accordance with bioanalytical validation guidelines. The described analytical method was proven to be robust and was successfully applied to the determination of fentanyl in plasma and CSF samples from a pharmacokinetic and pharmacodynamic study in newborn piglets receiving intravenous fentanyl (5 µg/kg bolus immediately followed by a 90-min infusion of 3 µg/kg/h).

  9. Rapid determination of piracetam in human plasma and cerebrospinal fluid by micellar electrokinetic chromatography with sample direct injection.

    Science.gov (United States)

    Yeh, Hsin-Hua; Yang, Yuan-Han; Ko, Ju-Yun; Chen, Su-Hwei

    2006-07-07

    A simple micellar electrokinetic chromatography (MEKC) method with UV detection at 200 nm for analysis of piracetam in plasma and in cerebrospinal fluid (CSF) by direct injection without any sample pretreatment is described. The separation of piracetam from biological matrix was performed at 25 degrees C using a background electrolyte consisting of Tris buffer with sodium dodecyl sulfate (SDS) as the electrolyte solution. Several parameters affecting the separation of the drug from biological matrix were studied, including the pH and concentrations of the Tris buffer and SDS. Under optimal MEKC condition, good separation with high efficiency and short analyses time is achieved. Using imidazole as an internal standard (IS), the linear ranges of the method for the determination of piracetam in plasma and in CSF were all between 5 and 500 microg/mL; the detection limit of the drug in plasma and in CSF (signal-to-noise ratio=3; injection 0.5 psi, 5s) was 1.0 microg/mL. The applicability of the proposed method for determination of piracetam in plasma and CSF collected after intravenous administration of 3g piracetam every 6h and oral administration 1.2g every 6h in encephalopathy patients with aphasia was demonstrated.

  10. Effect of a copper intrauterine contraceptive device and nylon suture on the estradiol 17beta-6, 7-H3 and progesterone 1, 2-H3 in the rat uterus.

    Science.gov (United States)

    Ghosh, M; Roy, S K; Kar, A B

    1975-01-01

    Copper wire or nylon suture was inserted in 1 horn of rats and their uterin were subsequently studied to determined the effect of these IUDs on in vitro estradiol-17beta, 6,7-tritiated and progesterone 1,2-tritiated. Control horns took up significancy more (p .01) estradiol than either copper or nylon IUD-fitted horns. The copper IUD horn showed an increase (p .01) in progesterone uptake while the nylon IUD horn showed a decrease (p .01) in progesterone uptake. The estrogen to progesterone ratio in uptake of the control and IUD horn of the nylon IUD-fitted rats had significantly less (p .01) uptake of both progesterone and estradiol compared to either copper or nylon IUD-fitted rats.

  11. Effect of antiprogestin ZK 98. 734 on the ovarian cycle, early pregnancy, and on its binding to progesterone receptors in the myometrium of marmoset Callithrix jacchus

    Energy Technology Data Exchange (ETDEWEB)

    Puri, C.P.; Kholkute, S.D.; Pongubala, J.M.; Patil, R.K.; Elger, W.A.; Jayaraman, S.

    1988-04-01

    The antiprogestin ZK 98.734 (11 beta-(4-dimethylaminophenyl-17 beta-hydroxy-17 alpha-(3-hydroxy-prop-1(Z)-enyl-4,9(10)-estradien-3-one) was administered i.m. (5 mg/day) for three consecutive days to two groups of common marmosets. In one group (nonpregnant, n = 6), it was injected during the luteal phase, and to the second group (pregnant, n = 7), it was injected during early pregnancy, on Days 24-26 of the mid-cycle estradiol peak. Administration of ZK 98.734 during the luteal phase caused a sharp drop in plasma progesterone levels. The luteal phase was shortened whether the drug was administered during the early or the late luteal phase. Similarly, administration of ZK 98.734 during early pregnancy caused a significant drop in progesterone levels, and pregnancy was terminated in all of the animals. The post-treatment cycles in both groups of animals were ovulatory and of normal duration. /sup 3/H-ZK 98.734 showed specific binding to myometrial cytosol fraction. ZK 98.734 also displaced the binding of /sup 3/H-progesterone to progesterone receptors. However, progesterone had higher binding affinity than did ZK 98.734. The antifertility action of ZK 98.734 could be a result either of its luteolytic action or of its blocking the progesterone receptors in the target tissue. This study, therefore, indicates that in the common marmoset ZK 98.734 is a progesterone antagonist with a potential to terminate early pregnancy.

  12. Determination of snake-necked turtle Hydromedusa tectifera (Cope, 1870) (Testudines: Chelidae) plasma protein concentrations by refractometry and the biuret method.

    OpenAIRE

    2011-01-01

    The purpose of this study was to evaluate the accuracy of hand-held refractometer in determining plasma protein concentrations in South American snake-necked turtle (Hydromedusa tectifera) as compared with the standard biuret method. The results indicated that plasma protein values may be accurately determined in snake- necked turtle with a hand-held refractometer.

  13. STOPPIT Baby Follow-up Study: the effect of prophylactic progesterone in twin pregnancy on childhood outcome.

    Directory of Open Access Journals (Sweden)

    Helen Christine McNamara

    Full Text Available To determine the long-term effects of in utero progesterone exposure in twin children.This study evaluated the health and developmental outcomes of all surviving children born to mothers who participated in a double-blind, placebo-controlled trial of progesterone given for the prevention of preterm birth in twin pregnancies (STOPPIT, ISRCTN35782581. Follow-up was performed via record linkage and two parent-completed validated questionnaires, the Child Development Inventory and the Health Utilities Index.Record linkage was successfully performed on at least one record in 759/781 (97% children eligible for follow-up. There were no differences between progesterone-exposed and placebo-exposed twins with respect to incidence of death, congenital anomalies and hospitalisation, nor on routine national child health assessments. Questionnaire responses were received for 324/738 (44% children. The mean age at questionnaire follow-up was 55.5 months. Delay in at least one developmental domain on the Child Development Inventory was observed in 107/324 (33% children, with no evidence of difference between progesterone-exposed and placebo-exposed twins. There was no evidence of difference between the progesterone and placebo groups in global health status assessed using the Health Utilities Index: 89% of children were rated as having 'excellent' health and a further 8% as having 'very good' health.In this cohort of twin children there was no evidence of a detrimental or beneficial impact on health and developmental outcomes at three to six years of age due to in utero exposure to progesterone.

  14. Interactions between inflammatory signals and the progesterone receptor in regulating gene expression in pregnant human uterine myocytes

    Science.gov (United States)

    Lee, Yun; Sooranna, Suren R; Terzidou, Vasso; Christian, Mark; Brosens, Jan; Huhtinen, Kaisa; Poutanen, Matti; Barton, Geraint; Johnson, Mark R; Bennett, Phillip R

    2012-01-01

    The absence of a fall in circulating progesterone levels has led to the concept that human labour is associated with ‘functional progesterone withdrawal’ caused through changes in the expression or function of progesterone receptor (PR). At the time of labour, the human uterus is heavily infiltrated with inflammatory cells, which release cytokines to create a ‘myometrial inflammation’ via NF-κB activation. The negative interaction between NF-κB and PR, may represent a mechanism to account for ‘functional progesterone withdrawal’ at term. Conversely, PR may act to inhibit NF-κB function and so play a role in inhibition of myometrial inflammation during pregnancy. To model this inter-relationship, we have used small interfering (si) RNA-mediated knock-down of PR in human pregnant myocytes and whole genome microarray analysis to identify genes regulated through PR. We then activated myometrial inflammation using IL-1β stimulation to determine the role of PR in myometrial inflammation regulation. Through PR-knock-down, we found that PR regulates gene networks involved in myometrial quiescence and extracellular matrix integrity. Activation of myometrial inflammation was found to antagonize PR-induced gene expression, of genes normally upregulated via PR. We found that PR does not play a role in repression of pro-inflammatory gene networks induced by IL-1β and that only MMP10 was significantly regulated in opposite directions by IL-1β and PR. We conclude that progesterone acting through PR does not generally inhibit myometrial inflammation. Activation of myometrial inflammation does cause ‘functional progesterone withdrawal’ but only in the context of genes normally upregulated via PR. PMID:22435466

  15. The role of progesterone in prevention of preterm birth

    Directory of Open Access Journals (Sweden)

    Jodie M Dodd

    2009-07-01

    Full Text Available Jodie M Dodd, Caroline A CrowtherDiscipline of Obstetrics and Gynaecology, The University of Adelaide, Adelaide, South Australia, AustraliaAbstract: Preterm birth continues to provide an enormous challenge in the delivery of perinatal health care, and is associated with considerable short and long-term health consequences for surviving infants. Progesterone has a role in maintaining pregnancy, by suppression of the calcium–calmodulin–myosin light chain kinase system. Additionally, progesterone has recognized anti-inflammatory properties, raising a possible link between inflammatory processes, alterations in progesterone receptor expression and the onset of preterm labor. Systematic reviews of randomized controlled trials evaluating the use of intramuscular and vaginal progesterone in women considered to be at increased risk of preterm birth have been published, with primary outcomes of perinatal death, preterm birth <34 weeks, and neurodevelopmental handicap in childhood. Eleven randomized controlled trials were included in the systematic review, involving 2714 women and 3452 infants, with results presented according to the reason women were considered to be at increased risk of preterm birth. While there is a potential beneficial effect in the use of progesterone for some women considered to be at increased risk of preterm birth, primarily in the reduction in the risk of preterm birth before 34 weeks gestation, it remains unclear if the observed prolongation of pregnancy translates into improved health outcomes for the infant.Keywords: progesterone, preterm birth, systematic review, randomized trial

  16. Progesterone Upregulates Gene Expression in Normal Human Thyroid Follicular Cells

    Directory of Open Access Journals (Sweden)

    Ana Paula Santin Bertoni

    2015-01-01

    Full Text Available Thyroid cancer and thyroid nodules are more prevalent in women than men, so female sex hormones may have an etiological role in these conditions. There are no data about direct effects of progesterone on thyroid cells, so the aim of the present study was to evaluate progesterone effects in the sodium-iodide symporter NIS, thyroglobulin TG, thyroperoxidase TPO, and KI-67 genes expression, in normal thyroid follicular cells, derived from human tissue. NIS, TG, TPO, and KI-67 mRNA expression increased significantly after TSH 20 μUI/mL, respectively: 2.08 times, P<0.0001; 2.39 times, P=0.01; 1.58 times, P=0.0003; and 1.87 times, P<0.0001. In thyroid cells treated with 20 μUI/mL TSH plus 10 nM progesterone, RNA expression of NIS, TG, and KI-67 genes increased, respectively: 1.78 times, P<0.0001; 1.75 times, P=0.037; and 1.95 times, P<0.0001, and TPO mRNA expression also increased, though not significantly (1.77 times, P=0.069. These effects were abolished by mifepristone, an antagonist of progesterone receptor, suggesting that genes involved in thyroid cell function and proliferation are upregulated by progesterone. This work provides evidence that progesterone has a direct effect on thyroid cells, upregulating genes involved in thyroid function and growth.

  17. [Selenium determination in plasma/serum by inductively coupled plasma mass spectrometry (ICP-MS): comparison with graphite furnace atomic absorption spectrometry (GF-AAS)].

    Science.gov (United States)

    Janasik, Beata; Trzcinka-Ochocka, Małgorzata; Brodzka, Renata

    2011-01-01

    The present study was aimed at comparing two techniques of selenium (Se) determination in serum/plasma samples: inductively coupled plasma mass spectrometry (ICP-MS) and graphite furnace atomic absorption (GF-AAS). Blood samples were collected by venipuncture, using Venosafe closed blood sampling system. The samples were centrifuged. The measurements were performed by Elan DRC-e mass spectrometry, Perkin Elmer, SCIEX, USA and Unicam Solar 989 QZ atomic absorption spectrometry. Reference material, Clincheck Serum Control Level 1 (Recipe, Germany), was used to verify the determinations. The Laboratory participates in external quality control (G-EQUAS). Analytical parameters for both techniques are respectively: ICP-MS--precision 5.9%, limit of detection 0.19 microg/l, repeatability 5.5%, trueness 2.4%, bias 97.6%, GF-AAS--precision 8%, limit of detection 3.4 microg/l, repeatability 7.2%, trueness 6.8%, bias 93.2%. The benefits of the ICP-MS technique are high accuracy, low detection limits and the possibility of multi-element analysis.

  18. Novel determinants of H-Ras plasma membrane localization and transformation

    DEFF Research Database (Denmark)

    Willumsen, B M; Cox, A D; Solski, P A;

    1996-01-01

    cysteine did not abolish palmitoylation. However, despite continued lipid modification the mutant proteins failed to bind to plasma membranes and instead accumulated on internal membranes and, importantly, were not transforming. Addition of an N-terminal myristoylation signal to these defective mutants......, or to proteins entirely lacking the C-terminal 25 residues restored both plasma membrane association and transforming activity. Thus, H-Ras does not absolutely require prenylation or palmitoylation nor indeed its hypervariable domain in order to interact with effectors that ultimately cause transformation....... However, in this native state, the C-terminus appears to provide a combination of lipids and a previously unrecognized signal for specific plasma membrane targeting that are essential for the correct localization and biological function of H-Ras....

  19. Liquid chromatography determination of clobazam and its major metabolite N-desmethylclobazam in human plasma.

    Science.gov (United States)

    Brachet-Liermain, A; Jarry, C; Faure, O; Guyot, M; Loiseau, P

    1982-01-01

    A specific procedure for the analysis of clobazam and N-desmethylclobazam in plasma is described. Reversed-phase liquid chromatography was performed on a Radial-pak cartridge using a mixture of 45% acetonitrile and 55% buffer solution (pH 7); the ultraviolet detector was set at 254 nm. The method used diazepam as internal standard and diethylether as extraction solvent. The calibration curves are linear between 50 and 500 ng/ml for clobazam and between 100 and 1000 ng/ml for N-desmethylclobazam. The day-to-day precision of the procedure at clobazam plasma concentrations of 50, 250, and 500 ng/ml generated coefficients of variation of 2.2, 6.6, and 11.3%, respectively. No interference occurred in plasma from patients treated with various drugs. The method has been used to study the pharmacokinetics of clobazam and N-desmethylclobazam in patients receiving oral clobazam.

  20. Determination of the Ion Velocity Distribution in a Rotating Plasma from Measurements of Doppler Broadening

    DEFF Research Database (Denmark)

    Jørgensen, L. W.; Sillesen, Alfred Hegaard

    1979-01-01

    The Doppler-broadened profile of the He II 4685.75 AA line was measured along a chord in a rotating plasma, transverse to the magnetic field. Using a single-particle orbit picture, the corresponding velocity spectrum of ions confirm the measurements, so it can be concluded that the single......-particle orbit picture is valid for the discharge period under investigation, except for the first few microseconds during breakdown when a strong interaction between plasma and remaining neutral gas takes place by Alfvens critical velocity mechanism. A simple relation is given between the measured half......-width and shift of the Doppler profile and the macroscopic quantities of ion velocity and energy. Several Doppler-broadened profiles are shown for different plasma parameters....

  1. Determination of elastic modulus and residual stress of plasma-sprayed tungsten coating on steel substrate

    Energy Technology Data Exchange (ETDEWEB)

    You, J.H. [Max-Planck-Institut fuer Plasmaphysik, EURATOM Association, Boltzmann Street 2, 85748 Garching (Germany)]. E-mail: jeong-ha.you@ipp.mpg.de; Hoeschen, T. [Max-Planck-Institut fuer Plasmaphysik, EURATOM Association, Boltzmann Street 2, 85748 Garching (Germany); Lindig, S. [Max-Planck-Institut fuer Plasmaphysik, EURATOM Association, Boltzmann Street 2, 85748 Garching (Germany)

    2006-01-01

    Plasma-sprayed tungsten, which is a candidate material for the first wall armour, shows a porous, heterogeneous microstructure. Due to its characteristic morphology, the properties are significantly different from those of its dense bulk material. Measurements of the elastic modulus of this coating have not been reported in the literature. In this work Young's modulus of highly porous plasma-sprayed tungsten coatings deposited on steel (F82H) substrates was measured. For the fabrication of the coating system the vacuum plasma-spray process was applied. Measurements were performed by means of three-point and four-point bending tests. The obtained modulus values ranged from 53 to 57 GPa. These values could be confirmed by the test result of a detached coating strip, which was 54 GPa. The applied methods produced consistent results regardless of testing configurations and specimen sizes. The errors were less than 1%. Residual stress of the coating was also estimated.

  2. Determination of elastic modulus and residual stress of plasma-sprayed tungsten coating on steel substrate

    Science.gov (United States)

    You, J. H.; Höschen, T.; Lindig, S.

    2006-01-01

    Plasma-sprayed tungsten, which is a candidate material for the first wall armour, shows a porous, heterogeneous microstructure. Due to its characteristic morphology, the properties are significantly different from those of its dense bulk material. Measurements of the elastic modulus of this coating have not been reported in the literature. In this work Young's modulus of highly porous plasma-sprayed tungsten coatings deposited on steel (F82H) substrates was measured. For the fabrication of the coating system the vacuum plasma-spray process was applied. Measurements were performed by means of three-point and four-point bending tests. The obtained modulus values ranged from 53 to 57 GPa. These values could be confirmed by the test result of a detached coating strip, which was 54 GPa. The applied methods produced consistent results regardless of testing configurations and specimen sizes. The errors were less than 1%. Residual stress of the coating was also estimated.

  3. Progesterone status, parity, body condition, and days postpartum before estrus or ovulation synchronization in suckled beef cattle influence artificial insemination pregnancy outcomes.

    Science.gov (United States)

    Stevenson, J S; Hill, S L; Bridges, G A; Larson, J E; Lamb, G C

    2015-05-01

    Our objective was to assess the effects of progesterone before initiating an estrus- or ovulation-synchronization program in addition to the influence of parity, BCS, and days postpartum on resulting pregnancy rates per AI. Experimental data were combined from 73 herd-year studies consisting of more than 8,500 suckled beef cows exposed to variants of the CO-Synch program. Blood was harvested from samples collected at 10 and 0 d before the onset of CO-Synch, and progesterone concentrations of the samples were determined. The progesterone environment preceding synchronization was assessed in 3 ways on the basis of progesterone concentrations measured in the 2 defined blood samples. All binomial logistic regression models used procedure GLIMMIX in SAS and included the fixed effects of program duration, inclusion of progesterone via an intravaginal insert, parity, days postpartum at AI, BCS, and appropriate interactions. In addition, model 1 included 3 categories of progesterone concentrations (low [primiparous cows that began in a low-progesterone environment (proestrus, estrus, metestrus, or anestrus). A significant 3-way interaction of parity, BCS, and days postpartum in 2 models reinforced the importance of these factors to AI pregnancy outcomes. Ancillary analyses identified the significant effects of cycling status and BCS as well as days postpartum on luteolytic response to PGF(2α). Pregnancy loss of 2.7% to 4.2% was detected to occur between a positive pregnancy diagnosis at 35 d post-AI and later stages of pregnancy. We concluded that progesterone status at the onset of the synchronization program is critical to pregnancy outcomes in primiparous but not multiparous cows.

  4. Gas temperature determination in an argon non-thermal plasma at atmospheric pressure from broadenings of atomic emission lines

    Science.gov (United States)

    Yubero, C.; Rodero, A.; Dimitrijevic, M. S.; Gamero, A.; García, M. C.

    2017-03-01

    In this work a new spectroscopic method, allowing gas temperature determination in argon non-thermal plasmas sustained at atmospheric pressure, is presented. The method is based on the measurements of selected pairs of argon atomic lines (Ar I 603.2 nm/Ar I 549.6 nm, Ar I 603.2 nm/Ar I 522.1 nm, Ar I 549.6 nm/Ar I 522.1 nm). For gas temperature determination using the proposed method, there is no need of knowing the electron density, neither making assumptions on the degree of thermodynamic equilibrium existing in the plasma. The values of the temperatures obtained using this method, have been compared with the rotational temperatures derived from the OH ro-vibrational bands, using both, the well-known Boltzmann-plot technique and the best fitting to simulated ro-vibrational bands. A very good agreement has been found.

  5. A fast and sensitive method for the determination of nitrite in human plasma by capillary electrophoresis with fluorescence detection.

    Science.gov (United States)

    Wang, Xu; Adams, Erwin; Van Schepdael, Ann

    2012-08-15

    Analysis of nitrite, the indicator of nitric oxide (NO) generation in vivo, provides a useful tool to study NO synthesis in vivo. A fast and sensitive fluorometric CE method was developed for determination of nitrite in human plasma through its derivatization with 2,3-diaminonaphthalene (DAN). Nitrite in human plasma was easily reacted with DAN under acid conditions to yield the highly fluorescent 2,3-naphthotriazole (NAT). Fluorescence detection was optimized to achieve subnanomolar detection which allows a direct analysis of plasma samples unlike most CE-UV methods using sample stacking. Acetonitrile was used to remove the protein. Short-end injection and a high voltage (-30 kV) were used to shorten the analysis time. The good separation was achieved with 20 mM borate buffer at pH 9.23. The separation of NAT was obtained within 1.4 min. The deproteinized plasma sample was injected hydrodynamically for 5s at -50 mbar into a 60 cm × 75 μm internal diameter uncoated fused-silica capillary. Excitation wavelength was selected with a broad-band filter (240-400 nm), and the emitted light was measured at 418 nm by the use of a cutoff filter. A good linearity (R(2)=0.9975) was obtained in the range from 2 to 500 nM. The detection limit of nitrite was 0.6 nM in original plasma samples, which is 750 times lower than our previous CE-UV method. The developed fluorometric CE method offers the advantages of more simple system and lower cost compared with the current fluorometric HPLC methods without losing sensitivity. The detected mean nitrite concentration in human plasma by this method was consistent with the most frequently reported values.

  6. Simultaneous determination of oxysterols, cholesterol and 25-hydroxy-vitamin D3 in human plasma by LC-UV-MS.

    Directory of Open Access Journals (Sweden)

    Rohini Narayanaswamy

    Full Text Available Oxysterols are promising biomarkers of neurodegenerative diseases that are linked with cholesterol and vitamin D metabolism. There is an unmet need for methods capable of sensitive, and simultaneous quantitation of multiple oxysterols, vitamin D and cholesterol pathway biomarkers.A method for simultaneous determination of 5 major oxysterols, 25-hydroxy vitamin D3 and cholesterol in human plasma was developed. Total oxysterols were prepared by room temperature saponification followed by solid phase extraction from plasma spiked with deuterated internal standards. Oxysterols were resolved by reverse phase HPLC using a methanol/water/0.1% formic acid gradient. Oxysterols and 25-hydroxy vitamin D3 were detected with atmospheric pressure chemical ionization mass spectrometry in positive ion mode; in-series photodiode array detection at 204nm was used for cholesterol. Method validation studies were performed. Oxysterol levels in 220 plasma samples from healthy control subjects, multiple sclerosis and other neurological disorders patients were quantitated.Our method quantitated 5 oxysterols, cholesterol and 25-hydroxy vitamin D3 from 200 μL plasma in 35 minutes. Recoveries were >85% for all analytes and internal standards. The limits of detection were 3-10 ng/mL for oxysterols and 25-hydroxy vitamin D3 and 1 μg/mL for simultaneous detection of cholesterol. Analytical imprecision was <10 %CV for 24(S-, 25-, 27-, 7α-hydroxycholesterol (HC and cholesterol and ≤15 % for 7-keto-cholesterol. Multiple Sclerosis and other neurological disorder patients had lower 27-hydroxycholesterol levels compared to controls whereas 7α-hydroxycholesterol was lower specifically in Multiple Sclerosis.The method is suitable for measuring plasma oxysterols levels in human health and disease. Analysis of human plasma indicates that the oxysterol, bile acid precursors 7α-hydroxycholesterol and 27-hydroxycholesterol are lower in Multiple Sclerosis and may serve as potential

  7. Relaxin and progesterone during pregnancy and the post-partum period in association with live and stillborn calves in bottlenose dolphins (Tursiops truncatus).

    Science.gov (United States)

    Bergfelt, Don R; Steinetz, Bernard G; Lasano, Salamia; West, Kristi L; Campbell, Michelle; Adams, Gregg P

    2011-02-01

    The objectives of this study were to validate a relaxin and progesterone RIA for use in bottlenose dolphins, and quantify and characterize both hormones in extracts of placental tissue and serum collected during pregnancy and the post-partum period, and compare the results between dolphins with live and stillborn calves. In Experiment 1, validation of a heterologous relaxin and progesterone RIA involved specific displacement of antibody-bound radiolabeled human relaxin or progesterone in response to increasing volumes of pooled pregnant dolphin serum and amounts of respective hormone standards added to a fixed volume of serum. The displacement curves were considered parallel and additive relative to respective standard curves. In Experiment 2, immunoreactive relaxin and progesterone were detected in placental extracts and, in corresponding serum samples, concentrations of both hormones were higher during the pre-partum than post-partum periods. Circulatory concentrations of progesterone decreased (P interaction, P interaction, P = 0.17). Even though the interaction did not reach significance, mean relaxin concentrations were 42%, 29%, and 34% lower at early, mid-, and late pregnancy, respectively, in dolphins with stillbirths than in those with live births. In conclusion, the pregnancy-specific increase in serum concentrations of relaxin and lower concentrations of both relaxin and progesterone in association with stillbirths suggest the potential for relaxin to be used diagnostically to determine pregnancy status, and one or both hormones to be used to assess placental function, and, perhaps, fetal well-being in bottlenose dolphins and other cetaceans.

  8. A Re-Evaluation of Trapped Plasma in the Microhematocrit Determination.

    Science.gov (United States)

    1986-01-01

    reported that trapped plasma values were increased in samples of healthy subjects with hereditary spherocytosis . Economou-Marvou and Tsenghi in 1965 (9...TARGETS LF 2.50 31.9 83.8 28.1 33.5 13.8 *MOD ELLIPTOCYTOSIS SP 2.85 30.3 83.6 32.5 38.9 27.8 *MOD SPHEROCYTOSIS AN 2.09 35.2 82.7 26.5 32.1 15.6...lower red cell size. They stated that anisocytosis and spherocytosis by themselves do not alter the amount of trapped plasma, and that microcytic cells

  9. Synthesis of silicon carbide in a nitrogen plasma torch: rotational temperature determination and material analysis

    Energy Technology Data Exchange (ETDEWEB)

    Ruiz-Camacho, J; Castell, R [Universidad Simon BolIvar, Departamento de Fisica, Caracas (Venezuela, Bolivarian Republic of); Castro, A; Manrique, M [Universidad Simon BolIvar, Departamento de Ciencias de los Materiales, Caracas (Venezuela, Bolivarian Republic of)], E-mail: jgruiz@usb.ve

    2008-09-07

    Experiments on silicon carbide synthesis were performed using a dc nitrogen plasma torch. Measurements of rotational temperature of nitrogen molecules by emission spectroscopy were performed, based on the band (0, 1) of the first negative system of nitrogen N{sub 2}{sup +}(B{sup 2}{sigma}{sub u}{sup +}{yields}X{sup 2}{sigma}{sub g}{sup +}) for the R branch. Three different plasma torch powers were studied in order to optimize the production of silicon carbide with our experimental set-up. The synthesized products were characterized by x-ray diffraction, scanning electron microscopy and energy dispersive x-ray spectroscopy.

  10. Synthesis of silicon carbide in a nitrogen plasma torch: rotational temperature determination and material analysis

    Science.gov (United States)

    Ruiz-Camacho, J.; Castell, R.; Castro, A.; Manrique, M.

    2008-09-01

    Experiments on silicon carbide synthesis were performed using a dc nitrogen plasma torch. Measurements of rotational temperature of nitrogen molecules by emission spectroscopy were performed, based on the band (0, 1) of the first negative system of nitrogen N_2^+ (B\\,{}^2\\Sigma_u^+ \\to X\\,{}^2\\Sigma _g^+) for the R branch. Three different plasma torch powers were studied in order to optimize the production of silicon carbide with our experimental set-up. The synthesized products were characterized by x-ray diffraction, scanning electron microscopy and energy dispersive x-ray spectroscopy.

  11. Determination of albumin transport rate between plasma and peritoneal space in decompensated cirrhosis

    DEFF Research Database (Denmark)

    Ring-Larsen, H; Henriksen, Jens Henrik Sahl

    1984-01-01

    the abdominal puncture may lead to overestimation of TERperit.space, whereas systematic understimation seems less likely. This may besides differences in patient selection and unsteady state, account for the discrepancy between the present relatively low value and earlier reports on much higher values.......10-0.59). The transport rate of albumin from ascitic fluid back to plasma was measured in eight patients by plasma sampling after intraperitoneal injection of 131I-labelled serum albumin. After correction for tracer re-extravasation this back transport (median 0.31, range 0.07-0.44% IVM/h-1) was not significantly...

  12. Determination of desmethyldiazepam in plasma by electron-capture GLC: application to pharmacokinetic studies of clorazepate.

    Science.gov (United States)

    Greenblatt, D J

    1978-03-01

    Plasma desmethyldiazepam concentrations were quantitated by a rapid and sensitive technique using electron-capture GLC. Following addition of diazepam as the internal standard, plasma is extracted at physiological pH into benzene-isoamyl alcohol. The extract is evaporated to dryness and reconstituted with toluene-isoamyl alcohol prior to chromatography. Both diazepam and desmethyldiazepam are quantitatively extracted. The variation of identical samples is 5%, and the sensitivity is 5 ng of desmethyldiazepam/ml of original sample. The method is applicable to pharmacokinetic studies of clorazepate, a benzodiazepine derivative transformed to desmethyldiazepam prior to absorption.

  13. Pharmacokinetic Properties of Three Forms of Vaginal Progesterone Administered in Either Single Or Multiple Dose Regimen in Healthy Post-menopausal Chinese Women

    Directory of Open Access Journals (Sweden)

    Jianzhong Shentu

    2017-04-01

    Full Text Available Objective: A generic vaginal progesterone gel has recently been developed in China. Little is known about its pharmacokinetic properties in Chinese subjects. The purpose of our study was to investigate the pharmacokinetics of three forms of vaginal progesterone gel (test formulations at 4 and 8% strength vs. a reference formulation: Crinone 8% in Chinese healthy post-menopausal women.Methods: This study consisted of two parts study. The part 1 study was a single-center, open-label, 3-period study. Twelve healthy post-menopausal women were to evaluate the safety and pharmacokinetics of 45 mg vaginal progesterone gel (Test 4% following single dose and multiple doses administered once every other day (q.o.d. for six times or once daily (q.d. for 6 days. The part 2 study was a randomized, open-label, 3-stage crossover study. Twelve post-menopausal women received 90 mg vaginal progesterone gel (Test 8% or 90 mg Crinone (Reference 8% following single dose and multiple doses (q.o.d. or q.d.. Plasma concentrations of progesterone were measured up to 72 h by using a validated liquid chromatography tandem-mass spectrometry method. The primary pharmacokinetic parameters, maximum plasma concentration (Cmax and area under the plasma concentration–time curve (AUC from time zero to last measurable concentration (AUC0-t and extrapolated to infinity (AUC0-∞ were compared by an analysis of variance using log-transformed data.Results: Totally 24 subjects were enrolled in and completed the study. Following single dose, The geometric mean Cmax values for Test 4%, Test 8%, and Crinone 8% were 6.35, 10.34, 10.45 ng/mL, and their geometric mean AUC0-t (AUC0-∞ were 113.73 (118.00, 169.39 (173.98, and 190.07 (201.13 ng⋅h/mL, respectively. The mean T1/2 values of progesterone were 11.00, 10.92, and 11.40 h, respectively. For 8% test formulation vs. reference, the 90% CIs of the least squares mean test/reference ratios of Cmax, AUC0-t, and AUC0-∞ were 78.32–124

  14. A New Liquid Chromatography-Tandem Mass Spectrometry Method for Determination of Bisoprolol in Human Plasma Samples

    OpenAIRE

    Gabriela Peste; Nela Bibire; Mihai Apostu; Aurel Vlase; Corneliu Oniscu

    2009-01-01

    Liquid chromatography (LC) coupled with mass spectrometry (MS) detection is one of the most powerful analytical tools for organic compound analysis. The advantages of using LC/MS methods over HPLC methods include: selectivity, chromatographic integrity, peak assignment, structural information, and rapid method development. In this paper, a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the determination of bisoprolol in human plasma s...

  15. Application of inductively coupled plasma quadrupole mass spectrometry for the determination of monazite ages by lead isotope ratios

    OpenAIRE

    Godoy,José Marcus; Godoy,Maria Luiza D. P; Aronne,Cláudia C.

    2007-01-01

    In order to evaluate the applicability of inductively coupled plasma quadrupole mass spectrometry to the determination of Pb/Pb, U/Pb and Th/Pb ages of monazite, studies were carried out initially applying lead atom ratio reference standards (NIST SRM 981 and 982). Further, the optimized methodology was applied to monazite sands from three different sites, Sugar Loaf Hill (Rio de Janeiro city), Buena (Rio de Janeiro state) and Black Sands Beach (Guarapari, Espirito Santo state); the obtained ...

  16. Isolation of Francisella tularensis and Yersinia pestis from Blood Cultures by Plasma Purification and Immunomagnetic Separation Accelerates Antibiotic Susceptibility Determination

    Science.gov (United States)

    Aloni-Grinstein, Ronit; Schuster, Ofir; Yitzhaki, Shmuel; Aftalion, Moshe; Maoz, Sharon; Steinberger-Levy, Ida; Ber, Raphael

    2017-01-01

    The early symptoms of tularemia and plague, which are caused by Francisella tularensis and Yersinia pestis infection, respectively, are common to other illnesses, resulting in a low index of suspicion among clinicians. Moreover, because these diseases can be treated only with antibiotics, rapid isolation of the bacteria and antibiotic susceptibility testing (AST) are preferable. Blood cultures of patients may serve as a source for bacteria isolation. However, due to the slow growth rates of F. tularensis and Y. pestis on solid media, isolation by plating blood culture samples on proper agar plates may require several days. Thus, improving the isolation procedure prior to antibiotic susceptibility determination is a major clinically relevant need. In this study, we developed a rapid, selective procedure for the isolation of F. tularensis and Y. pestis from blood cultures. We examined drop-plating and plasma purification followed by immunomagnetic separation (IMS) as alternative isolation methods. We determined that replacing the classical isolation method with drop-plating is advantageous with respect to time at the expense of specificity. Hence, we also examined isolation by IMS. Sub-localization of F. tularensis within blood cultures of infected mice has revealed that the majority of the bacteria are located within the extracellular fraction, in the plasma. Y. pestis also resides within the plasma. Therefore, the plasma fraction was isolated from blood cultures and subjected to an IMS procedure using polyclonal anti-F. tularensis live vaccine strain (LVS) or anti-Y. pestis antibodies conjugated to 50-nm nano-beads. The time required to reach an inoculum of sufficient bacteria for AST was shortest when using the plasma and IMSs for both bacteria, saving up to 2 days of incubation for F. tularensis and 1 day for Y. pestis. Our isolation procedure provides a proof of concept for the clinical relevance of rapid isolation for AST from F. tularensis- and Y. pestis

  17. Kadar Estradiol dan Progesteron Serum, Tampilan Vulva dan Sitologi Apus Vagina Kambing Bligon Selama Siklus Birahi

    Directory of Open Access Journals (Sweden)

    Irkham Widiyono

    2012-11-01

    Full Text Available A study was carried out to determine the serum estradiol and progesterone concentration, vulvaappearance, and vaginal cytology during estrous cycle in bligon goat. Four female non pregnant bligongoats (Indonesian indigenous were used in this research. They were clinically healthy and showed normalestrous cycle. Estrous state was examined by using biological method. When the animals showed acceptanceto be mounted by a buck, they were confirmed as the first day of estrous cycle (estrous phase. Collectionof blood sample, preparation of vaginal smears, and measurement of some physical genital parameterswere conducted by standard veterinary clinical methods at day 11, day between 3-5, day between 6-16, andday between 17-19 of estrous cycle. Enzyme linked immunosorbent assay (ELISA was used to determineserum progesterone and estradiol concentrations. Mean difference of each parameter was analyzed byanalyses of variance (Anova, followed by Tuckey HSD. The serum estradiol concentration ranged between211.25-247.77 pg/ml. Peak of the estradiol was occurred at the first day of estrous phase, namely 247.77pg/ml and to decrease at day 3-16. The serum progesterone concentration was at the level of 0.21-0.70 ng/ml and showed a significant increase at day 6-16 (p<0.05. The specific physical genital changes (swellingvulva, reddish mucous membrane of vulva, and excretion of viscous transparent vaginal discharge wereidentified as the animals were at the estrous phase. Composition of exfoliative vaginal cytology did notshow any specific and consistent changes throughout the estrous cycle. Superficial and intermediate cellswere very dominant (>60% during estrous cycle. In conclusion, serum estradiol, progesterone, the vulvaappearance combined with sexual receptivity could be a good estrous predictor for female cycling bligongoat.

  18. Plasma interactions determine the composition in pulsed laser deposited thin films

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Jikun; Stender, Dieter; Conder, Kazimierz; Wokaun, Alexander; Schneider, Christof W.; Lippert, Thomas, E-mail: thomas.lippert@psi.ch [Paul Scherrer Institute, CH-5232 Villigen PSI (Switzerland); Döbeli, Max [Laboratory of Ion Beam Physics, ETH Zurich, CH-8093 Zurich (Switzerland)

    2014-09-15

    Plasma chemistry and scattering strongly affect the congruent, elemental transfer during pulsed laser deposition of target metal species in an oxygen atmosphere. Studying the plasma properties of La{sub 0.6}Sr{sub 0.4}MnO{sub 3}, we demonstrate for as grown La{sub 0.6}Sr{sub 0.4}MnO{sub 3-δ} films that a congruent transfer of metallic species is achieved in two pressure windows: ∼10{sup −3} mbar and ∼2 × 10{sup −1} mbar. In the intermediate pressure range, La{sub 0.6}Sr{sub 0.4}MnO{sub 3-δ} becomes cation deficient and simultaneously almost fully stoichiometric in oxygen. Important for thin film growth is the presence of negative atomic oxygen and under which conditions positive metal-oxygen ions are created in the plasma. This insight into the plasma chemistry shows why the pressure window to obtain films with a desired composition and crystalline structure is narrow and requires a careful adjustment of the process parameters.

  19. Determination of Arsenic in Sinus Wash and Tap Water by Inductively Coupled Plasma-Mass Spectrometry

    Science.gov (United States)

    Donnell, Anna M.; Nahan, Keaton; Holloway, Dawone; Vonderheide, Anne P.

    2016-01-01

    Arsenic is a toxic element to which humans are primarily exposed through food and water; it occurs as a result of human activities and naturally from the earth's crust. An experiment was developed for a senior level analytical laboratory utilizing an Inductively Coupled Plasma-Mass Spectrometer (ICP-MS) for the analysis of arsenic in household…

  20. Determination of trace elements in petroleum products by inductively coupled plasma techniques: A critical review

    Energy Technology Data Exchange (ETDEWEB)

    Sánchez, Raquel [Department of Analytical Chemistry, Nutrition and Food Sciences, P.O. Box 99, 03080, Alicante (Spain); Todolí, José Luis, E-mail: jose.todoli@ua.es [Department of Analytical Chemistry, Nutrition and Food Sciences, P.O. Box 99, 03080, Alicante (Spain); Lienemann, Charles-Philippe [IFP Energies Nouvelles, Rond-point de l' échangeur de Solaize, BP 3, F-69360 Solaize (France); Mermet, Jean-Michel [Spectroscopy Forever, 01390 Tramoyes (France)

    2013-10-01

    The fundamentals, applications and latter developments of petroleum products analysis through inductively coupled plasma optical emission spectrometry (ICP-OES) and mass spectrometry (ICP-MS) are revisited in the present bibliographic survey. Sample preparation procedures for the direct analysis of fuels by using liquid sample introduction systems are critically reviewed and compared. The most employed methods are sample dilution, emulsion or micro-emulsion preparation and sample decomposition. The first one is the most widely employed due to its simplicity. Once the sample has been prepared, an organic matrix is usually present. The performance of the sample introduction system (i.e., nebulizer and spray chamber) depends strongly upon the nature and properties of the solution finally obtained. Many different devices have been assayed and the obtained results are shown. Additionally, samples can be introduced into the plasma by using an electrothermal vaporization (ETV) device or a laser ablation system (LA). The recent results published in the literature showing the feasibility, advantages and drawbacks of latter alternatives are also described. Therefore, the main goal of the review is the discussion of the different approaches developed for the analysis of crude oil and its derivates by inductively coupled plasma (ICP) techniques. - Highlights: • Analysis of petroleum products by inductively coupled plasma techniques is revisited. • Fundamental studies are included together with reports dealing with applications. • Conventional and non-conventional sample introduction methods are considered. • Sample preparation methods are critically compared and described.

  1. Plasminogen activator inhibitor-type I: its plasma determinants and relation with cardiovascular risk

    NARCIS (Netherlands)

    Hoekstra, T.; Geleijnse, J.M.; Schouten, E.G.; Kluft, C.

    2004-01-01

    The habitual level of PAI-1 is influenced by many factors, of which obesity and insulin resistance are the most important. It is possible to reduce plasma PAI-1 by changes in life style, e.g. weight reduction and physical activity. Data on potential interactions between environmental and metabolic v

  2. A method for determination of dimethyl benzoylphenyl urea (BPU) in human plasma by using LC/UV.

    Science.gov (United States)

    Rudek, Michelle A; Zabelina, Yelena; Zhao, Ming; Wolff, Antonio C; Baker, Sharyn D

    2004-06-01

    Dimethyl benzoylphenyl urea (BPU) inhibited tubulin polymerization, caused microtubule depolymerization in vitro and demonstrated activity against solid tumors. BPU is being tested in phase I clinical trials. A rapid and specific method using LC/UV has been developed for quantitation of BPU in human heparin-containing plasma to perform pharmacokinetic and pharmacodynamic studies. BPU is extracted from plasma into acetonitrile:n-butyl-chloride using paclitaxel as the internal standard and separated on a Waters Symmetry C18 (3.9 x 150 mm, 5 microm) column with acetonitrile-water mobile phase (70:30, v/v) using isocratic flow at 1 mL/min for a run time of 5 min. Ultraviolet detection was utilized and performed at 225 nm for BPU and paclitaxel. The retention times were 1.9 min for paclitaxel and 4.1 min for BPU. Calibration curves were generated over the range of 0.01-10 microg/mL with coefficient of determination of > 0.99. The values for within-day and between-day precision were BPU 320 mg as a weekly oral dose to a patient with advanced solid tumor malignancies, the maximum plasma concentration was 2 micro g/mL and concentrations were quantifiable up to 168 h after administration. The lower limit of quantitation of 0.01 microg/mL allows for successful measurement of plasma concentrations in patients.

  3. [Deuterated surrogate and UPLC-MS/MS method for simultaneous determination of 22 endocannabinoids in ovariectomized rat plasma].

    Science.gov (United States)

    Xiang, Shi-Xie; Xu, Ying; Zhu, Jing-Jing; Wang, Zhi-Min; Zhang, Dong; Chen, Liang-Mian; Feng, Wei-Hong

    2016-11-01

    A new method based on ultraperformance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was developed for the simultaneous determination of 22 endocannabinoids(eCBs) and relevant compounds in ovariectomized rat plasma. After being extracted by solid-phase column(SPE), the plasma samples were detected by using UPLC-MS/MS. Analysis was carried out with ACQUITY UPLC BEH C₁₈ column. The mobile phase was 0.1% acetic acid solution(A)-acetonitrile and isopropanol(9∶1, B) for the gradient elution. In the positive ion multiple reaction monitoring(MRM) mode, deuterated reagents were taken as standard alternatives to calculate recoveries and simultaneously quantify 22 endocannabinoids. The established method provided a good linearity for the 22 eCBs, and their linearly dependent coefficients were all higher than 0.99. The limits of quantitation(LOQs) ranged from 0.089 6 to 1.965 2 nmol•L-1. Relative recoveries of 5 deuterated surrogates ranged between 11.40% and 129.9%. The repeatability study results showed that RSD was all less than 8.0%. The established method could be used to analyze PGF2a EA, AEA and other endogenous cannabinoids in plasma samples of ovariectomized rats. In summary, this method was proved to boast a high sensitivity, repeatability and practicability, and thus could be used in rat plasma lipid metabolomics study. Copyright© by the Chinese Pharmaceutical Association.

  4. Anisotropic Mechanical Properties of Plasma-Sprayed Thermal Barrier Coatings at High Temperature Determined by Ultrasonic Method

    Science.gov (United States)

    Wei, Qin; Zhu, Jianguo; Chen, Wei

    2016-02-01

    The mechanical properties of plasma-sprayed thermal barrier coatings (TBC) are of great scientific and technological significance for the design and fabrication of TBC systems. The ultrasonic method combined with a sing-around method for mechanical properties measurement of TBC is deduced and the elastic modulus can be determined in the spray, or longitudinal, direction, and the transverse direction. Tested specimens of plasma-sprayed TBC are detached from the substrate and treated with thermal exposure at 1400 °C. The elastic moduli along the longitudinal and transverse directions of the TBCs are measured by different types of ultrasonic waves combined with a sing-around method, while the Poisson's ratio is also obtained simultaneously. The experimental results indicate that the magnitude of longitudinal elastic modulus is larger than that of the transverse one, and thus the plasma-sprayed TBC has an anisotropic mechanical property. Moreover, the elastic moduli along both longitudinal and transverse directions change with high-temperature exposure time, which consists of a rapid increasing stage followed by a slow decreasing stage. In addition, the magnitude of Poisson's ratio increases slightly from 0.05 to 0.2 with the high-temperature exposure time. Generally, the microstructures in the plasma-sprayed coatings and their evolution in a high-temperature environment are the main causes of the varying anisotropic mechanical properties.

  5. Determination of salbutamol in human plasma and urine using liquid chromatography coupled to tandem mass spectrometry and its pharmacokinetic study.

    Science.gov (United States)

    Zhang, Dujuan; Teng, Yanni; Chen, Keguang; Liu, Sha; Wei, Chunmin; Wang, Benjie; Yuan, Guiyan; Zhang, Rui; Liu, Xiaoyan; Guo, Ruichen

    2012-10-01

    A sensitive and selective liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was developed and validated for the determination of salbutamol in human plasma and urine, and successfully applied to the pharmacokinetic study of salbutamol in Chinese healthy volunteers after inhalation of salbutamol sulfate aerosol. Salbutamol and the internal standard (IS) acetaminophen in plasma and urine were extracted with ethyl acetate, separated on a C(18) reversed-phase column, eluted with mobile phase of acetonitrile-ammonium acetate (5 m m; 30:70, v/v), ionized by positive ion pneumatically assisted electrospray and detected in the multi-reaction monitoring mode using precursor → product ions of m/z 240.2 → 148.1 for salbutamol and 152 → 110 for the IS. The lower limits of quantitation of salbutamol in human plasma and urine by this method were 0.02 and 1 ng/mL, respectively. The specificity, matrix effect, recovery, sensitivity, linearity, accuracy, precision and several stabilities were validated for salbutamol in human plasma and urine. In conclusion, the validation results showed that this method is robust, specific and sensitive, and can successfully fulfill the requirement of clinical pharmacokinetic study of salbutamol in healthy Chinese volunteers.

  6. Inductively coupled plasma-atomic emission spectroscopy: a computer controlled, scanning monochromator system for the rapid determination of the elements

    Energy Technology Data Exchange (ETDEWEB)

    Floyd, M.A.

    1980-03-01

    A computer controlled, scanning monochromator system specifically designed for the rapid, sequential determination of the elements is described. The monochromator is combined with an inductively coupled plasma excitation source so that elements at major, minor, trace, and ultratrace levels may be determined, in sequence, without changing experimental parameters other than the spectral line observed. A number of distinctive features not found in previously described versions are incorporated into the system here described. Performance characteristics of the entire system and several analytical applications are discussed.

  7. Uncertainty Estimation of Metals and Semimetals Determination in Wastewater by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES)

    Science.gov (United States)

    Marques, J. R.; Villa-Soares, S. M.; Stellato, T. B.; Silva, T. B. S. C.; Faustino, M. G.; Monteiro, L. R.; Pires, M. A. F.; Cotrim, M. E. B.

    2016-07-01

    The measurement uncertainty is a parameter that represents the dispersion of the results obtained by a method of analysis. The estimation of measurement uncertainty in the determination of metals and semimetals is important to compare the results with limits defined by environmental legislation and conclude if the analytes are meeting the requirements. Therefore, the aim of this paper is present all the steps followed to estimate the uncertainty of the determination of amount of metals and semimetals in wastewater by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). Measurement uncertainty obtained was between 4.6 and 12.2% in the concentration range of mg.L-1.

  8. RP-HPLC Determination of Three Anti-Hyperlipidemic Drugs in Spiked Human Plasma and in Dosage Forms

    Directory of Open Access Journals (Sweden)

    Ola. M. Abdallah

    2011-01-01

    Full Text Available Sensitive, simple and accurate high performance liquid chromatographic (HPLC methods for the determination of atorvastatin (AT, fluvastatin (FL and pravastatin (PV have been developed. The proposed methods involve the use of a 150 mmx4.6 mm Zorbax Extend-C18 column (5 μm particle size and different chromatographic conditions for the separation of the three statins. Linearity range was 5-40, 5-30 and 10-60 μg mL-1 for AT, FL and PV respectively. The developed methods proved to be successful in the determination of all studied drugs in spiked human plasma samples.

  9. The Effects of Progesterone on Oocyte Maturation and Embryo Development

    Directory of Open Access Journals (Sweden)

    Saeed Zavareh

    2013-01-01

    Full Text Available Oocyte maturation and embryo development are controlled by intra-ovarian factors suchas steroid hormones. Progesterone (P4 exists in the follicular fluid that contributes tonormal mammalian ovarian function and has several critical functions during embryodevelopment and implantation, including endometrial receptivity, embryonic survivalduring gestation and transformation of the endometrial stromal cells to decidual cells.It is well known that the physiological effects of P4 during the pre-implantation stages ofsome mammal’s embryos are mediated by P4 receptors and their gene expression is determined.The effects of P4 on oocytes and embryo development have been assessed bysome investigations, with contradictory results. P4, a dominant steroid in follicular fluidat approximately 18 hours after the luteinizing hormone (LH surge may have a criticalrole in maturation of oocytes at the germinal stage. However, it has been shown that differentconcentrations of P4 could not improve in vitro maturation rates of germinal vesicles(GV in cumulus oocyte complexes (COCs and cumulus denuded oocytes (CDOs.Culture media supplemented with P4 significantly improved mouse embryo development.In addition, an in vivo experimental design has shown high blastocyst survival andimplantation rates in P4-treated mice.In this review we explain some of the findings that pertain to the effects of P4 onoocyte maturation and embryo development both in vitro and in vivo.

  10. Expression of estrogen and progesterone receptors in subcutaneous endometriosis

    Directory of Open Access Journals (Sweden)

    Đorđević M.

    2010-01-01

    Full Text Available Endometriosis is a clinical disorder defined by the presence of functional endometrial tissue outside the uterine cavity. Depending on the localization of the endometrial tissue related to the pelvis, the endometriosis can be classified either as intrinsic or extrinsic. The prevalence of endometriosis is difficult to determine. Statistical data show that endometriosis could be associated both with female infertility (20% and pelvic pains (24%, while in 4.1% of affected women, endometriosis has asymptomatic forms. The total prevalence of endometriosis is estimated to be between 5-10%. A 35-year-old woman from Knic, Serbia, was admitted to the Obstetrics and Gynecology Clinic of the Clinical Center in Kragujevac for surgical treatment of a suspicious swelling in the pubic region. Following surgical intervention, a nut-sized tumor was removed and sent for both pathohistological and immunohistochemical analysis. The results confirmed the presence of subcutaneous endometriosis positive for both estrogen and progesterone receptors. Endometriosis is usually described as a steroid hormone-dependent change that resembles the eutopic endometrial tissue characteristic for the presence of both glandular and stromal tissues. Given the fact that endometrial lesions are estrogen-dependent tumors, a crucial factor in the development of endometriosis is a late exposure to the hormone, mostly estrogen. Spontaneous subcutaneous endometriosis is rarely observed, but it could be assumed if there is recurrent pelvic pain which intensifies during menstruation. Given the fact that endometriosis coexists with different autoimmune diseases, multidisciplinary approaches are required for its proper diagnosis. .

  11. Mercury determination in non- and biodegradable materials by cold vapor capacitively coupled plasma microtorch atomic emission spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Frentiu, Tiberiu, E-mail: ftibi@chem.ubbcluj.ro [Faculty of Chemistry and Chemical Engineering, Babes-Bolyai University, Arany Janos 11, 400028 Cluj-Napoca (Romania); Mihaltan, Alin I., E-mail: alinblaj2005@yahoo.com [National Institute for Research and Development of Optoelectronics Bucharest - Research Institute for Analytical Instrumentation, Donath 67, 400293 Cluj-Napoca (Romania); Ponta, Michaela, E-mail: mponta@chem.ubbcluj.ro [Faculty of Chemistry and Chemical Engineering, Babes-Bolyai University, Arany Janos 11, 400028 Cluj-Napoca (Romania); Darvasi, Eugen, E-mail: edarvasi@chem.ubbcluj.ro [Faculty of Chemistry and Chemical Engineering, Babes-Bolyai University, Arany Janos 11, 400028 Cluj-Napoca (Romania); Frentiu, Maria, E-mail: frentiu.maria@yahoo.com [National Institute for Research and Development of Optoelectronics Bucharest - Research Institute for Analytical Instrumentation, Donath 67, 400293 Cluj-Napoca (Romania); Cordos, Emil, E-mail: emilcordos@gmail.com [National Institute for Research and Development of Optoelectronics Bucharest - Research Institute for Analytical Instrumentation, Donath 67, 400293 Cluj-Napoca (Romania)

    2011-10-15

    Highlights: {yields} Use of a miniaturized analytical system with microtorch plasma for Hg determination. {yields} Determination of Hg in non- and biodegradable materials using cold vapor generation. {yields} Figures of merit and advantages of the miniaturized system for Hg determination. - Abstract: A new analytical system consisting of a low power capacitively coupled plasma microtorch (20 W, 13.56 MHz, 150 ml min{sup -1} Ar) and a microspectrometer was investigated for the Hg determination in non- and biodegradable materials by cold-vapor generation, using SnCl{sub 2} reductant, and atomic emission spectrometry. The investigated miniaturized system was used for Hg determination in recyclable plastics from electronic equipments and biodegradable materials (shopping bags of 98% biodegradable polyethylene and corn starch) with the advantages of easy operation and low analysis costs. Samples were mineralized in HNO{sub 3}-H{sub 2}SO{sub 4} mixture in a high-pressure microwave system. The detection limits of 0.05 ng ml{sup -1} or 0.08 {mu}g g{sup -1} in solid sample were compared with those reported for other analytical systems. The method precision was 1.5-9.4% for Hg levels of 1.37-13.9 mg kg{sup -1}, while recovery in two polyethylene certified reference materials in the range 98.7 {+-} 4.5% (95% confidence level).

  12. Progesterone increases brain-derived neuroptrophic factor expression and protects against glutamate toxicity in a mitogen-activated protein kinase- and phosphoinositide-3 kinase-dependent manner in cerebral cortical explants.

    Science.gov (United States)

    Kaur, Paramjit; Jodhka, Parmeet K; Underwood, Wendy A; Bowles, Courtney A; de Fiebre, Nancyellen C; de Fiebre, Christopher M; Singh, Meharvan

    2007-08-15

    The higher prevalence and risk for Alzheimer's disease in women relative to men has been partially attributed to the precipitous decline in gonadal hormone levels that occurs in women following the menopause. Although considerable attention has been focused on the consequence of estrogen loss, and thus estrogen's neuroprotective potential, it is important to recognize that the menopause results in a precipitous decline in progesterone levels as well. In fact, progesterone is neuroprotective, although the precise mechanisms involved remain unclear. Based on our previous observation that progesterone elicits the phosphorylation of ERK and Akt, key effectors of the neuroprotective mitogen-activated protein kinase (MAPK) and phosphoinositide-3 kinase (PI3-K) pathways, respectively, we determined whether activation of either of these pathways was necessary for progesterone-induced protection. With organotypic explants (slice culture) of the cerebral cortex, we found that progesterone protected against glutamate-induced toxicity. Furthermore, these protective effects were inhibited by either the MEK1/2 inhibitor UO126 or the PI3-K inhibitor LY294002, supporting the requirement for both the MAPK and PI3-K pathways in progesterone-induced protection. In addition, at a concentration and duration of treatment consistent with our neuroprotection data, progesterone also increased the expression of brain-derived neurotrophic factor (BDNF), at the level of both protein and mRNA. This induction of BDNF may be relevant to the protective effects of progesterone, in that inhibition of Trk signaling, with K252a, inhibited the protective effects of progesterone. Collectively, these data suggest that progesterone is protective via multiple and potentially related mechanisms. (c) 2007 Wiley-Liss, Inc.

  13. Pretreatment of plasma samples by a novel hollow fiber centrifugal ultrafiltrate device for the determination of cefaclor concentrations in human plasma.

    Science.gov (United States)

    Li, Jun-Mei; Li, Cen; Jiang, Ye; Ren, Shu-Meng

    2010-10-29

    A simple sample preparation method was developed by using a centrifugal ultrafiltration (CF-UF) device with hollow fiber (HF) for the determination of cefaclor in plasma by HPLC. Samples were placed into a homemade device, which was consisted of a glass tube and a U-shaped hollow fiber. The filtrate was withdrawn from the hollow fiber into a syringe after centrifugation and 20 μL was directly injected into the HPLC for analysis. The HPLC method had a linear calibration curve in the concentration range of 6.00×10(-2)-30.7 μg mL(-1)(r=0.9996). The limit of detection (LOD) and limit of quantitation (LOQ) were 0.02 and 0.06 μg mL(-1), respectively. The intra and inter-day precisions (RSD) were 1.7%, 1.2%, 1.0% and 3.6%, 2.5%, 1.9%, respectively, for three concentrations. Assay accuracy was higher than 99.2% and the absolute recovery was 86.8-92.5%. It is feasible to use this novel and low cost device for sample pretreatment for the analysis of cefaclor in plasma.

  14. Determination of scutellarin in plasma by solid phase extraction—high performance liquid chromatography and its pharmacokinetics