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Sample records for human follicular fluid

  1. Pollutants in human follicular fluid

    SciTech Connect

    Trapp, M.; Baukloh, V.; Bohnet, H.G.; Heeschen, W.

    1984-07-01

    The present study was initiated to clarify whether environmental pollutants can reach the human follicular fluid, an important biologic component for reproduction, in concentrations which could be harmful to the gamete. Fifteen patients with tubal malformations were treated in an in vitro fertilization program whereby 18 follicle aspirates were obtained for investigation. The substances which were measured in this study were alpha-, beta-, and gamma-hexachlorocyclohexane (HCH), dichlorodiphenyltrichlorethane (DDT), polychlorinated biphenyls (PCB), hexachlorobenzene (HCB), dieldrin, and heptachlorepoxide (HepE). None of the patients became pregnant during the treatment cycle.

  2. Melatonin in human preovulatory follicular fluid

    NASA Technical Reports Server (NTRS)

    Brzezinski, Amnon; Seibel, Machelle M.; Lynch, Harry J.; Deng, Mei-Hua; Wurtman, Richard J.

    1987-01-01

    Melatonin, the major hormone of the pineal gland, has antigonadotrophic activity in many mammals and may also be involved in human reproduction. Melatonin suppresses steroidogenesis by ovarian granulosa and luteal cells in vitro. To determine if melatonin is present in the human ovary, preovulatory follicular fluids (n = 32) from 15 women were assayed for melatonin by RIA after solvent extraction. The fluids were obtained by laparoscopy or sonographically controlled follicular puncture from infertile women undergoing in vitro fertilization and embryo transfer. All patients had received clomiphene citrate, human menopausal gonadotropin, and hCG to stimulate follicle formation. Blood samples were obtained by venipuncture 30 rain or less after follicular aspiration. All of the follicular fluids contained melatonim, in concentrations substantially higher than those in the corresponding serum. A positive correlation was found between follicular fluid and serum melatonin levels in each woman; these observations indicate that preovulatory follicles contain substantial amounts of melatonin that may affect ovarian steroidogenesis.

  3. Melatonin in human preovulatory follicular fluid

    NASA Technical Reports Server (NTRS)

    Brzezinski, Amnon; Seibel, Machelle M.; Lynch, Harry J.; Deng, Mei-Hua; Wurtman, Richard J.

    1987-01-01

    Melatonin, the major hormone of the pineal gland, has antigonadotrophic activity in many mammals and may also be involved in human reproduction. Melatonin suppresses steroidogenesis by ovarian granulosa and luteal cells in vitro. To determine if melatonin is present in the human ovary, preovulatory follicular fluids (n = 32) from 15 women were assayed for melatonin by RIA after solvent extraction. The fluids were obtained by laparoscopy or sonographically controlled follicular puncture from infertile women undergoing in vitro fertilization and embryo transfer. All patients had received clomiphene citrate, human menopausal gonadotropin, and hCG to stimulate follicle formation. Blood samples were obtained by venipuncture 30 min or less after follicular aspiration. All of the follicular fluids contained melatonin, in concentrations (35.6 plus or minus 4.8 (plus or minus SEM) pg/mL) substantially higher than those in the corresponding serum (10.0 plus or minus 1.4 pg/mL). A positive correlation was found between follicular fluid and serum melatonin levels in each woman (r = 0.770; P less than 0.001). These observations indicate that preovulatory follicles contain substantial amounts of melatonin that may affect ovarian steroidogenesis.

  4. Human follicular fluid adverses hamster spermatozoa motility.

    PubMed

    Wetzels, A; Goverde, H J; Bastiaans, L A; Rolland, R

    1989-01-01

    To determine the optimal conditions for in vitro spermatozoa vitality, human and hamster spermatozoa were incubated at 37 degrees C in T6 medium supplemented with different biologic fluids (10% v/v). The fluids tested were human serum (HUS), hamster serum (HAS), and human follicular fluid (HUF). After incubation the spermatozoa were investigated for their qualitative and quantitative motility. Human spermatozoa maintained a good vitality in all fluids tested (approximately 25% motility after 18-h incubation). The hamster spermatozoa had after an incubation of 4 h a motility of 28.4% in HUS, 14.2% in HAS, and 2.2% in HUF. The quality of the motility was also extremely low in HUF, whereas it was adequate in HUS and in HAS. The presence of species-specific substances in mammalian follicular fluid is discussed. PMID:2589906

  5. Human follicular fluid and effects on reproduction.

    PubMed

    Basuino, Laís; Silveira, Carolina F

    2016-01-01

    Fertility - the ability to produce offspring - is considered a prerequisite for the development and perpetuation of species. Several factors may positively or negatively affect one's reproductive capabilities, such as regular exercises and maintaining a healthy bodyweight, versus aging, obesity, and stress. Follicular fluid (FF) is a liquid composed primarily of hormones, enzymes, anticoagulants, electrolytes, reactive oxygen species and antioxidants, which fills the follicular antrum and acts as an important mediator in the communication between cells in the antral follicle while bathing and carrying nutrients to the oocyte. Thus, human FF is a key element to the success of natural fertilization present in every stage of the conception process, from the communication between gametes to the development of fully viable embryos, and a vital component in the occurrence of spontaneous pregnancies. This literature review aimed to describe the possible effects of human follicular fluid on the natural fertilization process and to assess its role in follicular growth, oocyte quality, sperm capacitation, fertilization, and early embryonic development. PMID:27203305

  6. Complement, complement activation and anaphylatoxins in human ovarian follicular fluid.

    PubMed Central

    Perricone, R; de Carolis, C; Moretti, C; Santuari, E; de Sanctis, G; Fontana, L

    1990-01-01

    Functionally active complement was sought and detected in human follicular fluids obtained during the pre-ovulatory period. All the functional complement activities tested, including total haemolytic complement, classical pathway activity and alternative pathway activity were present in nine fluids from four different donors with values within the normal serum range. The immunochemical analysis demonstrated the presence of complement factors from C1 to C9, of B and of C1 INH, H, I. Complement anaphylatoxins were found employing RIA techniques in amounts significantly higher than in human plasma, thus demonstrating that follicular fluid complement, at least during the pre-ovulatory period, is partially activated. A possible role for urokinase-like substances in such an activation was indicated by further in vitro experiments. The presence of active complement in follicular fluid can be relevant for the function of the enzymatic multi-factorial mechanism of ovulation. PMID:2242616

  7. Angiogenic, mitogenic, and chemotactic activity in human follicular fluid (HFF)

    SciTech Connect

    Bryant, S.M.; Frederick, J.L.; Gale, J.A.; Campeau, J.D.; diZerega, G.S.

    1986-03-01

    The capacity of human follicular fluid to induce neovascularization was investigated. Three parameters were employed to assess the extent of angiogenic activity: (1) new vessel formation on the chick chorioallantoic membrane (CAM); (2) mitogenesis and (3) chemotaxis of bovine aortic endothelial cells. HFF resuspended in hydron induced new blood vessel formation on the CAM, as manifested by a spoke-wheel pattern of vessels radiating from the locus of application after two to six days. Endothelial cells cultured with a 1:10 dilution of HFF for two days demonstrated an enhanced incorporation of /sup 3/H-thymidine into acid-precipitable material when compared to control cells. The ratio of counts-per-minute for HFF stimulated cells versus control cells was 3.02 +/- 0.53 (anti S.E.M., n = 5). Endothelial cells also exhibited a directional migration towards HFF through a polycarbonate membrane with 8..mu..m pores. The ratio of the number of cells migrating completely through the filter towards a 1:10 dilution of HFF compared to those migrating towards medium alone was 5.61 +/- 0.61 (anti +/- S.E.M., n = 3). Human serum at an equivalent protein concentration as HFF demonstrated no activity in the CAM, mitogenic, and chemotaxis assays. These results demonstrate specific angiogenic, mitogenic and chemotactic activity in human follicular fluid.

  8. Oxidative stress markers in preovulatory follicular fluid in humans.

    PubMed

    Jozwik, M; Wolczynski, S; Jozwik, M; Szamatowicz, M

    1999-05-01

    Intensified peroxidation in the Graafian follicle may be a factor compromising the normal development of the oocyte. The aim of this study was to measure concentrations of three oxidative stress markers: conjugated dienes, lipid hydroperoxides and thiobarbituric acid-reactive substances, in preovulatory follicular fluids and sera of 145 women attending an in-vitro fertilization programme, and to correlate these concentrations with pregnancy outcome. Determinations were conducted either with or without an antioxidant (10 microM butylated hydroxytoluene) and an iron chelate (10 microM deferoxamine mesylate) to examine peroxidation associated with the methods used. Concentrations of conjugated dienes, lipid hydroperoxides and thiobarbituric acid-reactive substances in follicular fluid were all significantly lower than those in serum, both in the presence or absence of the antioxidant and iron chelate. These concentrations did not correlate with pregnancy outcome. In conclusion, the intensity of peroxidation in the Graafian follicle is much lower than that in serum. This gradient is the result of the lower rate of initiation of peroxidation in the follicular fluid, suggestive of the presence of efficient antioxidant defence systems in the direct milieu of the oocyte before ovulation. The concentrations of investigated oxidative stress markers in follicular fluid do not reflect the reproductive potential of oocytes. PMID:10338363

  9. Identification of gonadotrophin surge-inhibiting factor (GnSIF)/attenuin in human follicular fluid.

    PubMed

    Mroueh, J M; Arbogast, L K; Fowler, P; Templeton, A A; Friedman, C I; Danforth, D R

    1996-03-01

    Evidence from several laboratories suggests that the ovaries of many species produce a non-steroidal factor called gonadotrophin surge-inhibiting or attenuating factor (GnSIF) which may regulate the response of the pituitary to gonadotrophin-releasing hormone (GnRH) and as such, may modulate the timing and/or amplitude of the luteinizing hormone (LH) surge. We have recently isolated a candidate GnSIF from porcine follicular fluid (PFF). Porcine GnSiF is a 69 kDa protein which has undetectable inhibin and follistatin immunological and biological activity. The present study was designed to purify and identify GnSIF from human follicular fluid. GnSIF activity was measured as suppression of GnRH-stimulated LH secretion from rat pituitary cells in primary culture. Human follicular fluid (approximately 500 ml) was recovered from patients undergoing in-vitro fertilization (IVF). GnSIF was purified by heparin-sepharose, Q-sepharose, S-sepharose, and hydrophobic interaction chromatography followed by isoelectric focusing. Gel electrophoresis and Western blot were used to identify human GnSIF and compare it with porcine GnSIF. Using these steps, we obtained a highly-purified preparation of GnSIF that manifests in-vitro bioactivity and chromatography characteristics similar to those observed for porcine GnSIF and that hybridizes with a porcine GnSIF antibody. Following treatment with human chorionic gonadotrophin/human menopausal gonadotrophin (HMG/HCG), human follicular fluid contained roughly 25% of the GnSIF (per mg protein) present in porcine follicular fluid. We conclude that GnSIF is present in human follicular fluid and may participate in the regulation of gonadotrophin secretion in this species. PMID:8671252

  10. Effect of addition of human follicular fluid on progesterone secretion by cultured sheep granulosa cells.

    PubMed

    Kumari, G L; Vohra, S; Raghavan, V

    1982-10-01

    The effect of addition of human follicular fluid to cultures of granulosa cells of large sheep follicles (4-6 mm in diameter) on basal and LH-stimulated progesterone secretion was investigated. Both luteinization and progesterone secretion were inhibited by addition of 10% (w/v) charcoal-treated follicular fluid from medium (2-6 mm) and large (7-16 mm) follicles which had low concentrations of estradiol-17 beta, progesterone and LH. In comparison, the fluid from large follicles, having high levels of the same hormones, stimulated both the parameters, and addition of LH along with the fluid had no further effect. Fluid collected from cystic follicles appeared to be stimulatory which also had elevated levels of estradiol-17 beta and progesterone. These findings indicate the presence of both the inhibitors and stimulators of luteinization in human follicular fluid. The effectiveness of any of them either to inhibit or stimulate luteinization probably will depend upon the composition of the follicular fluid and the stage of maturation of the follicles from which it was collected. PMID:6218983

  11. The effect of human follicular fluid on bovine oocyte developmental competence and embryo quality.

    PubMed

    Valckx, Sara D M; De Bie, Jessie; Michiels, Ellen D; Goovaerts, Ilse G; Punjabi, Usha; Ramos-Ibeas, Priscila; Gutierrez-Adan, Alfonso; Bols, Peter E; Leroy, Jo L

    2015-02-01

    In this study, the hypothesis that embryo development during routine IVF procedures is determined by the pre-ovulatory follicular fluid composition was tested. Follicular fluid from women with obesity ('obese') and a 'positive' or 'negative' IVF outcome was added during the in-vitro maturation of bovine oocytes. 'Negative' and 'obese' follicular fluid reduced bovine embryo development, compared with laboratory control embryo development (P < 0.05 or P < 0.1). The addition of follicular fluid also altered bovine blastocyst gene expression. Furthermore, LDHA and PPARGC1B gene expression differed between follicular fluid groups. Data suggest that pre-ovulatory follicular fluid can potentially affect oocyte developmental competence and embryo quality. Furthermore, the bovine model may be used as a screening tool. PMID:25498595

  12. Electron probe microanalysis of the chemical elemental content of human follicular fluid.

    PubMed

    Chong, A P; Taymor, M L; Lechene, C P

    1977-05-15

    Follicular fluid samples were obtained by puncturing follicles of ovaries in situ from patients undergoing laparotomy. Sodium, potassium, chloride, magnesium, calcium, phosphorus, and sulfur concentrations measured by electron probe microanalysis were similar to those of blood, with minimal differences. This suggested that culture media in which these electrolytes are added in concentrations similar to those of serum are appropriate for culture of the human oocyte. PMID:855870

  13. Granulosa Cell Apoptosis Induced by a Novel FSH Binding Inhibitory Peptide From Human Ovarian Follicular Fluid

    PubMed Central

    Chitnis, Swati S.; Navlakhe, Rajshri M.; Shinde, Gayatri C.; Barve, Sharmila J.; D'Souza, Serena; Mahale, Smita D.; Nandedkar, Tarala D.

    2008-01-01

    Pituitary gonadotropins, follicle-stimulating hormone and luteinizing hormone, are the key regulators of ovarian folliculogenesis; these are known to be directly or indirectly modulated by many intraovarian factors. Our group has identified and studied one such novel peptide from human ovarian follicular fluid. Its partial N-terminal eight amino acid sequence has been deduced, referred to as octapeptide (OP). OP induces follicular atresia in mice and interferes with normal ovarian function in non-human primates, this action being similar to the native peptide. Thus, in this study, an attempt has been made to elucidate the mechanism of action of the synthetic OP by studying the pathway of follicular atresia in mouse ovary. Changes in granulosa cells were studied using various apoptotic markers by flow cytometry and immunohistochemistry. An increase in apoptotic cell population in atretic- and peptide-treated groups was observed compared with normal controls. Interestingly, both these groups exhibited differences in the apoptotic pathway. Results showed that the mitochondrial pathway was predominant in the atretic group, whereas the Fas-FasL pathway was predominant in the peptide-treated groups. The ultrastructural study also showed apoptotic changes in the OP-treated and atretic groups; the pattern of apoptosis differed at the subcellular level. (J Histochem Cytochem 56:961–968, 2008) PMID:18645207

  14. Identification of a cAMP-dependent protein kinase in bovine and human follicular fluids.

    PubMed

    Yang, L S; Kadam, A L; Koide, S S

    1993-11-01

    A soluble protein kinase (PK) was purified from bovine and human follicular fluids (FF) by ultrafiltration through a PM-10 membrane followed by chromatography on heparin-agarose, DEAE-cellulose and cellulose phosphate columns. The PK phosphorylated calf thymus histones and endogenous FF proteins having estimated Mrs of 40, 62, 128 and 180 KD. cAMP enhanced PK activity; whereas protein kinase A (PKA)-inhibitor peptide blocked the activity. The present findings suggest that the enzyme is a cAMP-dependent PK. PMID:8118427

  15. Effects of human follicular fluid and high-density lipoproteins on early spermatozoa hyperactivation and cholesterol efflux

    PubMed Central

    Hamdi, Safouane M.; Vieitez, Gérard; Jaspard, Béatrice; Barbaras, Ronald; Perret, Bertrand; Mieusset, Roget; Parinaud, Jean; Collet, Xavier

    2010-01-01

    The preovulatory human follicular fluid contains only HDLs as a lipoprotein class with a typically high proportion of preβ HDL. We first examined the role of follicular fluid and HDL subfractions on human spermatozoa capacitation, a process characterized by a hyperactivation of the flagellar movement and a depletion of plasma membrane cholesterol. Whole follicular fluid and isolated HDL, used at constant free cholesterol concentration, were both able to promote an early flagellar hyperactivation. Moreover, incubation of [3H]cholesterol-labeled spermatozoa with follicular fluid induced a rapid cholesterol efflux from spermatozoa that was confirmed by mass measurements of cholesterol transfer. Using isolated HDL, the cholesterol efflux had a similar time course and represented 70% of that mediated by whole follicular fluid. We then analyzed the time course of radioactive labeling of HDL subfractions. In the first minute of incubation, we found that the preβ HDL fraction incorporated the main part of the radioactivity (60%), with the rest being found in α-HDL, but strikingly, the labeling of α-HDL increased with time at the expense of preβ HDL.Thus, our results indicate that HDLs are involved in both spermatozoa hyperactivation and cholesterol effl ux and suggest the role of preβ-HDL particles as fi rst cellular cholesterol acceptors. PMID:19965575

  16. Peritoneal fluid modifies the response of human spermatozoa to follicular fluid.

    PubMed

    Caille, Adriana M; Berta, Cesar L; Cuasnicú, Patricia S; Munuce, Maria Jose

    2012-04-01

    The aim of this study was to elucidate the mechanism involved in the acrosome reaction (AR) induced by follicular fluid (FF) in spermatozoa previously exposed to peritoneal fluid (PF). The influence of progesterone was also investigated. Semen samples were from 18 normozoospermic donors. PF samples were from 13 women with unexplained infertility and from a woman treated with synthetic progestagen. FF samples were collected from six women undergoing IVF/embryo transfer and pooled. Motile spermatozoa were capacitated overnight and a kinetic and inhibition study on the FF-induced AR was performed. Spermatozoa pretreated with PF were challenged with either FF or progesterone. The ability of progesterone- and progestagen-supplemented PF to induce AR was analysed. Enzyme-digested PF was also tested. Pre-incubation with PF for 60 min completely prevented the FF-induced AR; spermatozoa treated with PF were unable to respond to FF or progesterone and this effect was not reversible. Progesterone- and progestagen-supplemented PF stimulated the AR relative to controls. Enzyme-digested PF did not have an inhibitory capacity. These data strongly suggest that there are one or more inhibitory proteins in PF that interact with spermatozoa so as to prevent access of progesterone to its receptor and thus inhibit the occurrence of the AR. The oviduct, or Fallopian tube, provides a place for spermatozoa and egg transport and storage, fertilization and early embryo development. If ovulation has not occurred, spermatozoa may reside in the oviduct for several hours or even a few days, awaiting oocyte arrival. It is assumed that fluids present in the female genital tract may have a role in synchronizing the timing required to guarantee the success of fertilization. We previously observed that the peritoneal fluid that bathes the peritoneal cavity is a suitable medium for sperm survival and we also reported that this fluid could stabilize spermatozoa. In this study we show further evidence

  17. Proteomic analysis of mare follicular fluid during late follicle development

    PubMed Central

    2011-01-01

    Background Follicular fluid accumulates into the antrum of follicle from the early stage of follicle development. Studies on its components may contribute to a better understanding of the mechanisms underlying follicular development and oocyte quality. With this objective, we performed a proteomic analysis of mare follicular fluid. First, we hypothesized that proteins in follicular fluid may differ from those in the serum, and also may change during follicle development. Second, we used four different approaches of Immunodepletion and one enrichment method, in order to overcome the masking effect of high-abundance proteins present in the follicular fluid, and to identify those present in lower abundance. Finally, we compared our results with previous studies performed in mono-ovulant (human) and poly-ovulant (porcine and canine) species in an attempt to identify common and/or species-specific proteins. Methods Follicular fluid samples were collected from ovaries at three different stages of follicle development (early dominant, late dominant and preovulatory). Blood samples were also collected at each time. The proteomic analysis was carried out on crude, depleted and enriched follicular fluid by 2D-PAGE, 1D-PAGE and mass spectrometry. Results Total of 459 protein spots were visualized by 2D-PAGE of crude mare follicular fluid, with no difference among the three physiological stages. Thirty proteins were observed as differentially expressed between serum and follicular fluid. Enrichment method was found to be the most powerful method for detection and identification of low-abundance proteins from follicular fluid. Actually, we were able to identify 18 proteins in the crude follicular fluid, and as many as 113 in the enriched follicular fluid. Inhibins and a few other proteins involved in reproduction could only be identified after enrichment of follicular fluid, demonstrating the power of the method used. The comparison of proteins found in mare follicular fluid

  18. Detection of the metabolites of human plasma and follicular fluid in IVF-ET with microextraction and LC-TOF-MS.

    PubMed

    Wang, Gang; Yao, Shun; Liang, Xin; Zuo, Tao; Zhu, Minghui

    2015-01-01

    The metabolites from human plasma and follicular fluid in IVF-ET are related with the metabolic activity of follicular cells and further with oocyte quality. With liquid chromatography-time-of-flight-mass spectrometry (LC-TOF-MS) and organic extraction from plasma and follicular fluid, a fast and efficient method was established for detecting the metabolites of human plasma and follicular fluid. Certain metabolites were closely related with human physiological activities, such as hormone, amino acid, estrogen and fatty acid, and estrogen fatty acyllipid lipid were successfully detected in human plasma and follicular fluid. The metabolites in follicular fluid and plasma on the day of oocyte retrieval were found to be similar. But those in the latter and basal plasma were partially different. Methyl (3α ,5β ,7α ,12α)-3,7,12-trihydroxycholan-24-oate, androst-4-ene-3,17-dione and tryptophan were identified and might be potential biomarkers on oocyte quality. It demonstrates that LC-TOF-MS combined with organic extraction could be an efficient tool to investigate the metabolites of human plasma and follicular fluid. Based on this study, it is expected to provide the basis for the further steps to explore possible influential factors on oocyte quality. PMID:26410324

  19. Pretreatment of sperm with human follicular fluid for borderline male infertility.

    PubMed

    Blumenfeld, Z; Nahhas, F

    1989-05-01

    To test the usefulness of human follicular fluid (FF) in treating male infertility, we incubated washed sperm specimens from 31 couples undergoing intrauterine insemination (IUI), for male and/or unexplained infertility, with either FF or Ham's F-10 medium (Gibco, Grand Island, NY), in alternating cycles in a randomized manner. Semen specimens from 28 men were incubated with either medium or FF. Incubations with FF have increased sperm penetration assay (SPA) scores from 24.8 +/- 17.3 to 34.3 +/- 13.6 (P less than 0.01). Incubation with heat-inactivized FF also has increased SPA scores, although to a lesser extent than noninactivized FF. Seventeen pregnancies occurred in the 31 couples treated by IUI (54.8%), 16 of them in FF-treated cycles (51.6%) and one in "control" IUI cycles (3.2%, P less than 0.01). All pregnancies occurred within four treatment cycles. Thus, IUI after sperm wash and preincubation with FF may be suggested for four to six cycles to couples with male factor and/or unexplained infertility who are reluctant to resort to artificial insemination by donor or adoption, before attempting the more costly and complex in vitro fertilization-embryo transfer procedure. PMID:2707464

  20. Protein pathways working in human follicular fluid: the future for tailored IVF?

    PubMed

    Bianchi, Laura; Gagliardi, Assunta; Landi, Claudia; Focarelli, Riccardo; De Leo, Vincenzo; Luddi, Alice; Bini, Luca; Piomboni, Paola

    2016-01-01

    The human follicular fluid (HFF) contains molecules and proteins that may affect follicle growth, oocyte maturation and competence acquiring. Despite the numerous studies, an integrated broad overview on biomolecular and patho/physiological processes that are proved or supposed to take place in HFF during folliculogenesis and oocyte development is still missing. In this review we report, for the first time, all the proteins unambiguously detected in HFF and, applying DAVID (Database for Annotation, Visualization and Integrated Discovery) and MetaCore bioinformatic resources, we shed new lights on their functional correlation, delineating protein patterns and pathways with reasonable potentialities for oocyte quality estimation in in vitro fertilisation (IVF) programs. Performing a rigorous PubMed search, we redacted a list of 617 unique proteins unambiguously-annotated as HFF components. Their functional processing suggested the occurrence in HFF of a tight and highly dynamic functional-network, which is balanced by specific effectors, primarily involved in extracellular matrix degradation and remodelling, inflammation and coagulation. Metalloproteinases, thrombin and vitamin-D-receptor/retinoid-X-receptor-alpha resulted as the main key factors in the nets and their differential activity may be indicative of ovarian health and oocyte quality. Despite future accurate clinical investigations are absolutely needed, the present analysis may provide a starting point for more accurate oocyte quality estimation and for defining personalised therapies in reproductive medicine. PMID:27149979

  1. Human testis cytosol and ovarian follicular fluid contain high amounts of interleukin-1-like factor(s).

    PubMed

    Khan, S A; Schmidt, K; Hallin, P; Di Pauli, R; De Geyter, C; Nieschlag, E

    1988-08-01

    Human testicular cytosol and ovarian follicular fluid were analyzed for the presence of interleukin-1 (IL-1)-like factors. Both the follicular fluid and testis cytosol preparations exhibited significant IL-1-like activity as determined by the murine thymocyte proliferation bioassay. The dose-response lines obtained with the gonadal preparations were parallel to each other and to those obtained with monocyte-derived IL-1 and the activity of the gonadal IL-1 could be neutralized by specific IL-1 antibodies. After gel chromatography of human follicular fluid (hFF) and human testis cytosol (hTC) proteins, IL-1 activity was found in the molecular weight region between 30 and 50 kilodaltons (kDa). Chromatofocusing of IL-1 from hFF and hTC revealed that the major part of IL-1 in both cases exhibited similar charge properties (pI less than 6.0). However, two extra peaks (pI 7.0 and greater than 9.0, respectively) were observed in hFF preparations. After isoelectrofocusing (IEF), IL-1 activity of hFF was also found in two different pH regions; a broad area of activity was localized between pH 5.5 and 7.0, while a sharp peak was observed with an approximate pI value of 9.5. Re-chromatofocusing or IEF of alkaline IL-1-like activity resulted in a heterogeneous profile of IL-1-like activity suggesting that the alkaline material may represent either a precursor or an aggregated form of the acidic IL-1. None of the IL-1 peaks obtained from hFF or hTC exhibited IL-2 activity as assessed in a specific IL-2 bioassay. The results of the present study indicate that both gonads may produce high amounts of IL-1-like factor(s) which might play a regulatory role in normal gonadal function. PMID:3264798

  2. MiRNA-320 in the human follicular fluid is associated with embryo quality in vivo and affects mouse embryonic development in vitro

    PubMed Central

    Feng, Ruizhi; Sang, Qing; Zhu, Yan; Fu, Wei; Liu, Miao; Xu, Yan; Shi, Huijuan; Xu, Yao; Qu, Ronggui; Chai, Renjie; Shao, Ruijin; Jin, Li; He, Lin; Sun, Xiaoxi; Wang, Lei

    2015-01-01

    Previous work from our laboratory demonstrated the existence of miRNAs in human follicular fluid. In the current study, we have sought to identify miRNAs that might affect oocyte/embryo quality in patients undergoing intracytoplasmic sperm injection and to investigate their roles in in vitro fertilization outcomes in mouse oocytes. 53 samples were classified as Group 1 (high quality) if the day-3 embryos had seven and more cells or as Group 2 (low quality) if the embryos had six and fewer cells. TaqMan Human microRNAs cards and qRT-PCR were performed to verify differently expressed miRNAs. The function of the corresponding miRNA was investigated in mouse oocytes by injecting them with miRNA-inhibitor oligonucleotides. We found that hsa-miR-320a and hsa-miR-197 had significantly higher expression levels in the Group 1 follicular fluids than in Group 2 (p = 0.0073 and p = 0.008, respectively). Knockdown of mmu-miR-320 in mouse oocytes strongly decreased the proportions of MII oocytes that developed into two-cell and blastocyst stage embryos (p = 0.0048 and p = 0.0069, respectively). Wnt signaling pathway components had abnormal expression level in miR-320 inhibitor-injected oocytes. This study provides the first evidence that miRNAs in human follicular fluid are indicative of and can influence embryo quality. PMID:25732513

  3. MiRNA-320 in the human follicular fluid is associated with embryo quality in vivo and affects mouse embryonic development in vitro.

    PubMed

    Feng, Ruizhi; Sang, Qing; Zhu, Yan; Fu, Wei; Liu, Miao; Xu, Yan; Shi, Huijuan; Xu, Yao; Qu, Ronggui; Chai, Renjie; Shao, Ruijin; Jin, Li; He, Lin; Sun, Xiaoxi; Wang, Lei

    2015-01-01

    Previous work from our laboratory demonstrated the existence of miRNAs in human follicular fluid. In the current study, we have sought to identify miRNAs that might affect oocyte/embryo quality in patients undergoing intracytoplasmic sperm injection and to investigate their roles in in vitro fertilization outcomes in mouse oocytes. 53 samples were classified as Group 1 (high quality) if the day-3 embryos had seven and more cells or as Group 2 (low quality) if the embryos had six and fewer cells. TaqMan Human microRNAs cards and qRT-PCR were performed to verify differently expressed miRNAs. The function of the corresponding miRNA was investigated in mouse oocytes by injecting them with miRNA-inhibitor oligonucleotides. We found that hsa-miR-320a and hsa-miR-197 had significantly higher expression levels in the Group 1 follicular fluids than in Group 2 (p = 0.0073 and p = 0.008, respectively). Knockdown of mmu-miR-320 in mouse oocytes strongly decreased the proportions of MII oocytes that developed into two-cell and blastocyst stage embryos (p = 0.0048 and p = 0.0069, respectively). Wnt signaling pathway components had abnormal expression level in miR-320 inhibitor-injected oocytes. This study provides the first evidence that miRNAs in human follicular fluid are indicative of and can influence embryo quality. PMID:25732513

  4. A Mixture Reflecting Polybrominated Diphenyl Ether (PBDE) Profiles Detected in Human Follicular Fluid Significantly Affects Steroidogenesis and Induces Oxidative Stress in a Female Human Granulosa Cell Line.

    PubMed

    Lefevre, Pavine L C; Wade, Mike; Goodyer, Cindy; Hales, Barbara F; Robaire, Bernard

    2016-07-01

    Brominated flame retardants are incorporated into consumer products to prevent flame propagation. These compounds leach into the domestic environment, resulting in chronic exposure. Pregnancy failure is associated with high levels of polybrominated diphenyl ethers (PBDEs), a major class of brominated flame retardants, in human follicular fluid, raising serious questions regarding their impact on female fertility. Our goal was to elucidate the effects of a mixture of PBDEs, similar to the profile found in human follicular fluid, on an immortalized human granulosa cell line, the KGN cell line. We showed that cell viability was altered and oxidative stress was induced as reflected by increased reactive oxygen species formation at 100 μM of the PBDE mixture. Transcriptomic analysis revealed that PBDE treatments of 1, 5, and 20 μM altered the expression of several genes involved in the reactive oxygen species signaling pathway. Significant dose-dependent reductions in progesterone and estradiol levels in the culture medium were measured after PBDE treatment; in parallel, the expression of genes involved in estradiol metabolism, namely CYP1A1, was up-regulated by 5 and 20 μM of the PBDE mixture. Treatment with 20 μM PBDE also increased the expression and secretion of the proinflammatory factor, IL-6, into the KGN cell culture medium. Our results demonstrate that PBDEs can alter human granulosa cell functions by inducing oxidative stress and disrupting steroidogenesis. These results indicate that PBDEs may be detrimental to ovarian functions and thus may adversely affect female reproductive health after chronic exposure. PMID:27219277

  5. A Proteomic Analysis of Human Follicular Fluid: Comparison between Younger and Older Women with Normal FSH Levels

    PubMed Central

    Hashemitabar, Mahmoud; Bahmanzadeh, Maryam; Mostafaie, Ali; Orazizadeh, Mahmoud; Farimani, Marzieh; Nikbakht, Roshan

    2014-01-01

    The follicular fluid (FF) is produced during folliculogenesis and contains a variety of proteins that play important roles in follicle development and oocyte maturation. Age-related infertility is usually considered as a problem that can be solved by assisted reproduction technology. Therefore, the identification of novel biomarkers that are linked to reproductive aging is the subject of this study. FF was obtained from healthy younger (20–32 years old) and older (38–42 years old) women undergoing intracytoplasmic sperm injection (ICSI) due to male factor infertility. The FF was analyzed by two-dimensional gel electrophoresis (2-DE). The power of two-dimensional gel electrophoresis and the identification of proteins were exploited using matrix-assisted laser desorption-ionization time-of-flight/time-of-flight (MALDI-TOF-TOF) mass spectrometry. Twenty three protein spots showed reproducible and significant changes in the aged compared to the young group. Of these, 19 protein spots could be identified using MALDI-TOF-TOF-MS. As a result of MASCOT search, five unique downregulated proteins were identified in the older group. These were identified as serotransferrin, hemopexin precursor, complement C3, C4 and kininogen. A number of protein markers were found that may help develop diagnostic methods of infertility. PMID:25268621

  6. Variability in the components of high density lipoprotein particles measured in human ovarian follicular fluid - A cross-sectional analysis

    PubMed Central

    Bloom, Michael S.; Kim, Keewan; Fujimoto, Victor Y.; Browne, Richard W.

    2014-01-01

    Objective Assess the variability of follicular fluid (FF) high-density lipoprotein (HDL) particle components. Design FF specimens were collected from two contralateral follicles on the day of oocyte retrieval and analyzed for HDL components. We characterized analytes by age, body mass index (BMI), race and smoking using a cross-sectional design. Biological variability was assessed using two-stage nested analysis of variance. Setting Reproductive health center. Patients One-hundred eighty in vitro fertilization (IVF) patients. Interventions None. Main Outcome Measures Nineteen HDL components including HDL-cholesterol and free (unesterified) and esterified forms, phospholipids, triglycerides, apolipoproteins A-1 and A-2, paraoxonase 1 (PON1) activities, and seven lipophilic vitamins and micronutrients. Results For some analytes, a majority of total measurement variability was attributed to sources between-follicles, suggesting an important role for the integrity of the blood-follicle-barrier and in situ remodeling of plasma derived constituents. For other analytes, variability was mostly attributed to sources between-women, likely indicative of plasma levels. Variability between-follicles decreased with increasing age, differed by BMI and smoking, and generally were lower for Asians and women with diminished ovarian reserve. Conclusions Substantial variability in FF HDL components exist between-follicles among women undergoing IVF, as well as between-women by age, BMI, race, smoking and by infertility diagnosis. PMID:24581578

  7. Effect of Follicular Fluid and Platelet-Activating Factor on Lactate Dehydrogenase C Expression in Human Asthenozoospermic Samples

    PubMed Central

    Esmaeilpour, Tahereh; Zarei, Mohmmad-Reza; Bahmanpour, Soghra; Aliabadi, Elham; Hosseini, Ahmad; Jaberipour, Mansooreh

    2014-01-01

    Background: Application of follicular fluid (FF) and platelet-activating factor (PAF) in artificial insemination improves sperm motility. Lactate dehydrogenase C (LDH-C) is a key enzyme for sperm motility. In this study, the effects of FF and PAF on the sperm motility index and LDH-C expression were investigated. Moreover, LDH-C expression was compared between asthenozoospermic and normozoospermic samples. Methods: The expression of LDH-C was examined by quantitative real-time polymerase chain reaction (q-RT PCR) and western blotting after it was treated with optimized concentrations of FF and PAF in twenty asthenozoospermic samples. Also, LDH-C expression was evaluated in five normozoospermic samples. Results: Samples with 75% FF and 100 nM of PAF had an increase in their percentages of progressive and slowly motile sperms and a decrease in their percentages of non-progressive and non-motile sperms. Moreover, LDH-C mRNA transcripts were not changed following PAF and FF treatment, and LDH-C protein was detected in highly progressive motile specimens treated with FF in the asthenozoospermic samples. Furthermore, LDH-C expression was more detectable in the normal sperms. Conclusion: Our results indicated that PAF had more beneficial effects than FF on sperm motility in the asthenozoospermic samples (P=0.0001), although the LDH-C expressions of the sperms were not changed significantly in both groups. We found no association between LDH-C expression and sperm motility after FF and PAF actions. This finding, however, requires further investigation. The fact that LDH-C protein was detected in the normozoospermic, but not asthenozoospermic, samples could be cited as a reason for the infertility in these patients. PMID:24453390

  8. Elemental composition of rabbit antral fluid during preovulatory follicular swelling.

    PubMed

    Burgoyne, P S; Borland, R M; Biggers, J D; Lechene, C P

    1979-11-01

    Electron probe microanalysis was used to determine the concentrations of Na, Cl, K Ca, Mg, S and P in samples of follicular fluid, ovarian vein serum and peripheral venous serum obtained from virgin rabbits at 2-h intervals up to 10 h after injection of hCG. Throughout this 10-h period the elemental composition of follicular fluid was essentially the same as that of blood serum. However, there was a significant drop in follicular fluid Ca relative to blood during the 10-h period which may reflect Ca involvement in the regulation of oocyte maturation. Significant differences were also found between follicles within rabbits for K and P concentrations. PMID:513038

  9. Serum allotypes in ovarian follicular fluids of pigs.

    PubMed

    Matousek, J; Hojný, J; Janik, A

    1986-01-01

    Sera and ovarian follicular fluids of 158 sows were tested with 27 allotype reagents. Immunodiffusion in agar gel (microtest) and haemagglutination inhibition were used as detection methods. Out of eight 'individual' (Lpb 1,-2,-3,-4,-5,-6,-7,-9) and four 'common' (Lpb 12,-13,-14,-16) specificities of serum beta-lipoproteins (LDL), 11 were present in sera, but none in follicular fluids. On the other hand, Lpr 1 and Lpr (x) allotypes of the VHDL + VLDL beta-lipoprotein system were detected both in sera and in follicular fluids. Of four antigens of the Gp system (Gp A,-a, -B,-b), only the 'dominant' characters, Gp A and Gp B, occurred in the follicular fluid. The typing of polymorphic IgG immunoglobulins (IgG-a or IgG-b system) showed that B1 or A2, B2 or A1 and B3 or A(x) allotypes could be detected both in serum and follicular fluid. Among allotypes that were not yet genetically classified, only the P3 specificity was not found in the population tested. The G1 allotype (preliminarily described as an alpha-globulin) was present in sera only, and the remaining allotypes, G9, P1, P16 and P23 (alpha- or beta-globulins) were present both in sera and follicular fluids. The mechanism of the transmission of serum proteins into ovarian follicles and their possible importance is discussed. PMID:3740552

  10. A comparison of the biochemical composition of equine follicular fluid and serum at four different stages of the follicular cycle.

    PubMed

    Collins, A; Palmer, E; Bézard, J; Burke, J; Duchamp, G; Buckley, T

    1997-12-01

    Samples of blood and follicular fluid were recovered from 27 Welsh Pony mares at 4 distinct stages of follicular development. Eighteen biochemical parameters were measured in each sample, including sodium, potassium, chloride, glucose, urea, creatinine, calcium, inorganic phosphate, total bilirubin, total protein, albumin, magnesium, triglyceride, total cholesterol, nonesterified fatty acids, alkaline phosphatase, gamma-glutamyltransferase and aspartate aminotransferase. The concentrations of progesterone, 17beta oestradiol and testosterone, pH and osmolarity, were also measured in all the follicular fluid samples. The concentrations of all proteins measured were lower in follicular fluid than serum whereas the reverse was true in the case of the lipids. Analysis of variance indicated that serum and follicular fluid concentrations of most of the parameters measured varied in parallel. PMID:9593520

  11. Metabolomic profiling of human follicular fluid from patients with repeated failure of in vitro fertilization using gas chromatography/mass spectrometry

    PubMed Central

    Xia, Lan; Zhao, Xiaoming; Sun, Yun; Hong, Yan; Gao, Yuping; Hu, Shuanggang

    2014-01-01

    Objective: To establish a gas chromatography/mass spectrometry (GC/MS)-based metabolomics method to compare the metabolites in the follicular fluid (FF) from patients with in vitro fertilization (IVF) and repeated IVF failure (RIF). Methods: A prospective study was employed in Center for Reprodutive Medcine, Renji Hospital, Shanghai, China, between January and October 2010. FF samples were collected from 13 patients with RIF and 15 patients who achieved pregnancy after the first IVF cycle. Results: Partial least squares (PLS) discriminant analysis of the PCA data revealed that the samples were scattered into two different regions. FF from the two groups differed with respect to 20 metabolites. FF from RIF group showed elevated levels of several amino acids (valine, threonine, isoleucine, cysteine, serine, proline, alanine, phenylalanine, lysine, methionine and ornithine), and reduced levels of dicarboxylic acids, cholesterol and some organic acids. Conclusions: The studies corroborated successful determination of the levels of metabolite in the FF. PMID:25400819

  12. Comparative Proteomic Analysis of Yak Follicular Fluid during Estrus

    PubMed Central

    Guo, Xian; Pei, Jie; Ding, Xuezhi; Chu, Min; Bao, Pengjia; Wu, Xiaoyun; Liang, Chunnian; Yan, Ping

    2016-01-01

    The breeding of yaks is highly seasonal, there are many crucial proteins involved in the reproduction control program, especially in follicular development. In order to isolate differential proteins between mature and immature follicular fluid (FF) of yak, the FF from yak follicles with different sizes were sampled respectively, and two-dimensional gel electrophoresis (2-DE) of the proteins was carried out. After silver staining, the Image Master 2D platinum software was used for protein analysis and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was performed for differential protein identification. The expression level of transferrin and enolase superfamily member 1 (ENOSF1) was determined by Western blotting for verification analysis. The results showed that 2-DE obtained an electrophoresis map of proteins from mature and immature yak FF with high resolution and repeatability. A comparison of protein profiles identified 12 differently expressed proteins, out of which 10 of them were upregulated while 2 were downregulated. Western blotting showed that the expression of transferrin and ENOSF1 was enhanced with follicular development. Both the obtained protein profiles and the differently expressed proteins identified in this study provided experimental data related to follicular development during yak breeding seasons. This study also laid the foundation for understanding the microenvironment during oocyte development. PMID:26954118

  13. Serum and follicular fluid concentrations of polybrominated diphenyl ethers and in-vitro fertilization outcome

    PubMed Central

    Johnson, Paula I.; Altshul, Larisa; Cramer, Daniel W.; Missmer, Stacey A.; Hauser, Russ; Meeker, John D.

    2012-01-01

    There is evidence of endocrine disruption and reproductive effects in animals following exposure to certain PBDEs, but human studies are limited. The goal of this study was to investigate the use of serum and follicular fluid as biomarkers of exposure to PBDEs and to explore whether a relationship between PBDE exposure and early pregnancy loss exists. We measured 8 PBDE congeners in archived serum and ovarian follicular fluid samples from 65 women undergoing in-vitro fertilization (IVF). Logistic regression models were used to predict the odds of failed embryo implantation associated with higher levels of PBDEs among the women in the study. There were moderate Kendall’s Tau-beta correlations between serum and follicular fluid concentrations of BDE 28, 47, 100 and 154 (Tβ =0.29–0.38, all p-values<0.005), but BDE 99 and 153 were not correlated between the two matrices (Tβ<0.2, p-values>0.05). Women with detectable concentrations of BDE 153 (39% had detectable levels) in follicular fluid had elevated odds of failed implantation compared with women who had non-detectable concentrations (adjusted OR=10.0; 95%CI: 1.9 to 52; p=0.006; adjusted by age and body mass index). These findings suggest that exposure to BDE 153 may be associated with failed embryo implantation. Due to our observation of only moderate correlations between matrices, serum PBDE concentrations may not be a good indicator of follicular fluid concentrations when studying early pregnancy endpoints in women undergoing IVF. PMID:22572111

  14. Do analogues of gonadotrophin releasing hormone influence follicular fluid steroid levels, oocyte maturity and fertilization rates?

    PubMed

    Tavmergen, E; Tavmergen, E N; Capanoğlu, R

    1992-04-01

    One-hundred-and-twelve samples of follicular fluid from 32 patients undergoing in-vitro fertilization and embryo transfer were analysed in this study. The follicular fluids were analysed for any relationships between oestradiol, progesterone and 17 alpha-hydroxyprogesterone levels, the progesterone/oestradiol and 17 alpha-hydroxyprogesterone/oestradiol ratios and oocyte maturity and fertilization rates. In Group A, consisting of women who used analogues of gonadotrophin-releasing hormone during ovarian stimulation with human menopausal gonadotrophin, the progesterone/oestradiol ratio rose in parallel with the fertilization rate (P less than 0.05). Group B comprised patients treated with human menopausal gonadotrophin alone. No significant relationship was found between the other parameters, oocyte maturation and fertilization rates in either group. PMID:1387881

  15. Immunohistochemical expression of MMP-14 and MMP-2, and MMP-2 activity during human ovarian follicular development

    PubMed Central

    2014-01-01

    Background The aim of this study was to investigate the presence of MMP-14 and MMP-2 during human ovarian follicular development using immunohistochemistry, and the activity of MMP-2 in follicular fluid using zymography. Methods Ovarian tissue collected from the archives of the Department of Pathology was examined and medical records and histopathology were reviewed. Follicular fluids were collected at the IVF-department and analyzed using zymography. Results MMP-14 and MMP-2 were increasingly found in the growing follicles and MMP-2 was highly expressed in the corpus luteum. Pro-MMP-2 was present in follicular fluid of IVF-patients. Conclusions The presence of MMP-14 and MMP-2 during human ovarian follicular development from the primordial follicle to the tertiary follicle and corpus luteum is confirmed, as was indicated by earlier animal studies following stimulation with gonadotrophins. PMID:24485069

  16. The porcine ovarian follicle: I. Selected chemical analysis of follicular fluid at different developmental stages.

    PubMed

    Chang, S C; Jones, J D; Ellefson, R D; Ryan, R J

    1976-10-01

    Numerous parameters of the chemical composition of porcine follicular fluid, obtained at different stages of follicle development, were analyzed. The concentrations of electrolytes, glucose, uric acid, selected enzymes, lipids, amino acids, cyclic AMP, and steroid hormones were determined and compared with concentrations in serum. There were both quantitative and qualitative differences in the composition of follicular fluid and plasma. These differences are discussed with regard to follicular development. PMID:183842

  17. Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence

    PubMed Central

    Sohel, Md. Mahmodul Hasan; Hoelker, Michael; Noferesti, Sina Seifi; Salilew-Wondim, Dessie; Tholen, Ernst; Looft, Christian; Rings, Franca; Uddin, Muhammad Jasim; Spencer, Thomas E.; Schellander, Karl; Tesfaye, Dawit

    2013-01-01

    Cell-cell communication within the follicle involves many signaling molecules, and this process may be mediated by secretion and uptake of exosomes that contain several bioactive molecules including extra-cellular miRNAs. Follicular fluid and cells from individual follicles of cattle were grouped based on Brilliant Cresyl Blue (BCB) staining of the corresponding oocytes. Both Exoquick precipitation and differential ultracentrifugation were used to separate the exosome and non-exosomal fraction of follicular fluid. Following miRNA isolation from both fractions, the human miRCURY LNA™ Universal RT miRNA PCR array system was used to profile miRNA expression. This analysis found that miRNAs were present in both exosomal and non-exosomal fraction of bovine follicular fluid. We found 25 miRNAs differentially expressed (16 up and 9 down) in exosomes and 30 miRNAs differentially expressed (21 up and 9 down) in non-exosomal fraction of follicular fluid in comparison of BCB- versus BCB+ oocyte groups. Expression of selected miRNAs was detected in theca, granulosa and cumulus oocyte complex. To further explore the potential roles of these follicular fluid derived extra-cellular miRNAs, the potential target genes were predicted, and functional annotation and pathway analysis revealed most of these pathways are known regulators of follicular development and oocyte growth. In order to validate exosome mediated cell-cell communication within follicular microenvironment, we demonstrated uptake of exosomes and resulting increase of endogenous miRNA level and subsequent alteration of mRNA levels in follicular cells in vitro. This study demonstrates for the first time, the presence of exosome or non-exosome mediated transfer of miRNA in the bovine follicular fluid, and oocyte growth dependent variation in extra-cellular miRNA signatures in the follicular environment. PMID:24223816

  18. Possible contribution of follicular interleukin-1beta to nitric oxide generation in human pre-ovulatory follicles.

    PubMed

    Tao, M; Kodama, H; Kagabu, S; Fukuda, J; Murata, M; Shimizu, Y; Hirano, H; Tanaka, T

    1997-10-01

    The aim of this study was to investigate the relationships between follicular nitric oxide (NO) metabolite concentrations and several related variables, with special reference to follicular interleukin-1beta (IL-1beta). The follicular fluid from the leading and secondary follicles was collected individually from 20 women undergoing in-vitro fertilization (IVF) treatment, and the concentrations of nitrite (NO2-) and nitrate (NO3-) were determined fluorometrically using 2,3-diaminonaphthalene. Both follicular nitrite (r = 0.42, P < 0.01) and nitrate (r = 0.49, P < 0.001) were found to be significantly correlated with follicular IL-1beta concentrations. There were also significant positive correlations between follicular nitrate and the number of oocytes retrieved (P < 0.01) and serum oestradiol concentration on the day of human chorionic gonadotrophin (HCG) administration (P < 0.05). When follicular cells were incubated in vitro with 10 ng/ml of IL-1beta for 24 h, nitrate generation was significantly (P < 0.01) elevated compared with the control. In conclusion, our study demonstrates that follicular IL-1beta and the number of developing follicles are significant variables that affect follicular NO concentrations, and points to the possible contribution of IL-1beta to NO generation in human preovulatory follicles. PMID:9402285

  19. Follicular fluid total antioxidant capacity levels in PCOS.

    PubMed

    Yilmaz, Nafiye; Inal, Hasan Ali; Gorkem, Umit; Sargin Oruc, Ayla; Yilmaz, Saynur; Turkkani, Ayten

    2016-07-01

    In this study, our aim was to assess total antioxidant capacity (TAC) levels in follicular fluid (FF) and their relationship to clinical pregnancy rates in PCOS patients undergoing assisted reproduction (ART). Twenty-two women with polycystic ovary syndrome (PCOS) (Group 1) and 41 women without PCOS (Group 2) were included in this study. Clinical and laboratory parameters and FF TAC levels were investigated. No statistically significant differences were found between the groups with regard to age and baseline parameters. Although we could not demonstrate a significant difference in FF TAC levels between the two groups (p=0.469), there was a significant positive correlation between FF TAC and clinical pregnancy rates, BMI, and the duration of infertility for the entire group (r=0.254, p=0.048; r=0.312, p=0.013; r=0.259, p=0.040; respectively). Owing to the correlation between FF TAC and the clinical pregnancy rates, further studies evaluating the impact of FF TAC levels on ART outcomes in patients with PCOS and other etiologies of infertility are needed. PMID:26911305

  20. Organochlorine pesticides in follicular fluid of women undergoing assisted reproductive technologies from central China.

    PubMed

    Zhu, Yindi; Huang, Bo; Li, Qing X; Wang, Jun

    2015-12-01

    Female infertility rates have increased by approximately 4% since the 1980s. There is evidence of adverse effects on female fertility in relation to exposure of chemical pollution in recent years. Follicular fluid samples were collected from 127 woman patients (aged 20-35) who underwent assisted reproductive technologies (ART) and had no records indicating occupational exposure to OCPs. Seventeen OCPs were analyzed in this study. The results showed that methoxychlor was dominant, accounted for 13.4% of total OCPs with a mean concentration of 167.9 ± 33.9 ng/g lipid weight (lw), followed by heptachlor-epoxide, hexachlorocyclohexanes, endrin and DDT. The concentrations of OCPs in the follicular fluid samples in the present study were moderate in comparison with those reported from developed or industrialized countries. All these pollutants can accumulate in different tissues of human body through diet, drinking water and respiration. No correlation between patient age and OCP concentrations was observed in this study. PMID:26412266

  1. Lipopolysaccharide in ovarian follicular fluid influences the steroid production in large follicles of dairy cows.

    PubMed

    Magata, Fumie; Horiuchi, Maya; Echizenya, Riku; Miura, Ryotaro; Chiba, Shiori; Matsui, Motozumi; Miyamoto, Akio; Kobayashi, Yoshiyasu; Shimizu, Takashi

    2014-01-10

    In postpartum dairy cows, various inflammatory diseases depress reproductive performance. Lipopolysaccharide (LPS) derived from infections of the uterus or mammary gland with Gram-negative bacteria was shown to suppress steroid production in the granulosa cells of follicles in vitro. The aim of the study was to investigate the relationship between LPS in ovarian follicular fluid and steroidogenesis by the theca and granulosa cells of the large follicles in vivo. Bovine ovaries were collected from a slaughterhouse, and the largest (F1) and the second largest (F2) follicles were used (>8 mm in diameter, n=38). LPS concentration in the follicular fluid was measured using quantitative kinetic assay. Follicular steroidogenesis was evaluated by measuring the estradiol (E2) and progesterone (P4) concentration in follicular fluid and by analysing transcription levels of steroidogenesis-related genes in theca and granulosa cells. LPS concentration detected in follicular fluid ranged from 0.2 to 2.0 EU/mL. In follicles with a high level of LPS (>0.5 EU/mL, n=15), the concentration of E2 was lower and that of P4 was higher when compared to those in follicles with a low level of LPS (<0.5 EU/mL, n=23), which was observed both in F1 and F2 follicles. Furthermore, in follicles with a high level of LPS, transcripts of steroidogenic enzymes such as CYP17 and P450arom were lower. In those follicles, the expression of caspase-3 was high, suggesting an association with follicular atresia. These findings indicate that LPS present in follicular fluid may cause ovarian dysfunction by inhibiting follicular activity. PMID:24321186

  2. Altered theca and cumulus oocyte complex gene expression, follicular arrest and reduced fertility in cows with dominant follicle follicular fluid androgen excess

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To date, animal models with naturally occurring androgen excess have not been identified. Serendipitously, we discovered two subpopulations of cows with dramatically different follicular fluid androgen concentrations in dominant follicles within our research herd. In the cow, androstenedione is the...

  3. Biochemical composition of ovine follicular fluid in relation to follicle size.

    PubMed

    Nandi, S; Kumar, V Girish; Manjunatha, B M; Gupta, P S P

    2007-01-01

    The aim of this study was to biochemically characterize ovine follicular fluid and to relate possible changes in composition to follicular size. Ovaries were collected from adult and cycling non-pregnant slaughtered sheep (Ovis aries) during breeding season. A total of 104 pairs of ovaries were investigated and these data were then compared. Follicular fluid was aspirated from small (< 2 mm), medium (2-4 mm) and large (> 4 mm) nonatretic ovarian follicles. The follicular fluid was centrifuged at 4 degrees C and 5000 g for 30 min to remove any cells and stored at -80 degrees C prior to assay. Follicular fluid samples were analyzed for glucose, total protein, cholesterol, triglycerides, lactate, urea, creatinine, sodium, potassium, chloride, calcium, phosphorus, magnesium, acid phosphatase, alkaline phosphatase, and lactate dehydrogenase. Data were analyzed by the linear regression model. As follicles became larger, the concentrations of glucose and cholesterol significantly (P < 0.05) increased while those of triglycerides, lactate, alkaline phosphatase and lactate dehydrogenase significantly (P < 0.05) decreased. PMID:17227345

  4. Profiling of bile acids in bovine follicular fluid by fused-core-LC-MS/MS.

    PubMed

    Sánchez-Guijo, A; Blaschka, C; Hartmann, M F; Wrenzycki, C; Wudy, S A

    2016-09-01

    Bile acids (BAs) are present in follicular fluid (FF) from humans and cattle. This fact has triggered an interest on the role BAs might play in folliculogenesis and their possible association with fertility. To achieve a better understanding about this subject, new methods are needed to provide reliable information about concentrations of the most important BAs in FF. In this context, liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers high specificity with a relatively simple sample workup. We developed and validated a new assay for the quick profiling of the 9 most abundant BAs in follicular fluid from cattle. The method uses 200μl of FF and can quantify cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA) and their glycine (G) and taurine (T) conjugates. Lithocholic acid (LCA), its conjugates GLCA and TLCA, and sulfated forms, were present in some samples, but their concentration was low compared to other BAs (in average, below 60ng/ml for LCA, GLCA or TLCA and below 20ng/ml for their corresponding sulfates). Method performance was studied at three quality controls for each compound in consonance with their physiological concentration. Excellent linearity and recovery were found for all compounds at every control level. Intra-day and between-day precisions (%CV) and accuracies (relative errors) were below 15% for all the compounds. Matrix effects were negligible for most of the analytes. Samples undergoing freeze-thaw showed no degradation of their BAs. The method makes use of a fused-core phenyl column coupled to a triple quadrupole tandem mass spectrometer to achieve chromatographic separation within 5min. We quantified BAs grouped in four different follicle sizes (3-5mm, 6-8mm, 9-14mm, >15mm), obtaining a similar relative BA profile for all the sizes, with CA always in higher concentration, ranging between 1600 and 18000ng/ml, approximately, followed by its conjugate glycocholic acid, GCA, which ranged between 800 and 9000ng

  5. The correlation of interleukin 1 and tumour necrosis factor to oestradiol, progesterone and testosterone levels in periovulatory follicular fluid of in-vitro fertilization patients.

    PubMed

    Barak, V; Mordel, N; Holzer, H; Zajicek, G; Treves, A J; Laufer, N

    1992-04-01

    Data has accumulated suggesting reciprocity between cytokines and the reproductive system. The present study was performed in order to evaluate the correlation between interleukin 1 (IL-1) and tumour necrosis factor (TNF) concentrations in follicular fluid and its oestradiol, progesterone and testosterone levels. A total of 39 follicular fluid samples, from eight patients undergoing in-vitro fertilization and embryo transfer were evaluated. All of the patients were treated by a midluteal (long) protocol involving a gonadotrophin releasing hormone agonist (GnRHa) coupled with follicular phase human menopausal gonadotrophin. Mean levels in follicular fluid of IL-1, TNF, oestradiol, progesterone and testosterone were 1.58 +/- 0.42 fmol/0.1 ml, 4.69 +/- 4.18 pg/ml, 28.5 +/- 58.1 ng/ml, 2360.5 +/- 2846.3 ng/ml and 7.22 +/- 7.08 ng/ml respectively. There was a significant (P less than 0.01) positive correlation between IL-1 and progesterone levels. There was no significant correlation between the different lymphokines and oestradiol secretion, oocyte fertilization, embryo quality and pregnancy rates. It is concluded that IL-1 and TNF exist in follicular fluid. It may be hypothesized that IL-1 has a local regulatory action, possibly promoting luteinization. PMID:1522186

  6. Dynamics of follicular fluid in one-humped camel (Camelus dromedarius).

    PubMed

    Zia-Ur-Rahman; Bukhari, S A; Ahmad, N; Akhtar, N; Ijaz, A; Yousaf, M S; Haq, I U

    2008-12-01

    In the present study, ovarian follicular fluid and serum biochemical, hormonal, electrolytes and amino acids profiles in female dromedary camel (Camelus dromedarius), were investigated. Fluid from small (2-6 mm) and large follicles (7-20 mm) and blood samples were collected from 25 clinically healthy adult female camels. The concentrations of glucose, cholesterol, triglycerides, high-density lipoproteins, urea, total proteins, albumin, globulin, fibrinogen, alanine aminotransferase, aspartate aminotransferase and tri-iodothyronine were lower (p < or = 0.05) in large follicles when compared with the small follicles. However, the concentrations of low-density lipoproteins, uric acid, creatinine, alkaline phosphatase and acid phosphatase in small and large follicles did not differ. The concentrations of oestradiol 17-beta and progesterone were higher (p < or = 0.05) in large follicles. The serum concentrations of these hormones were many folds lower (p < or = 0.05) than those of follicular fluid. Among electrolytes, the concentration of phosphorus was higher (p < or = 0.05) in the large follicles, while that of potassium and chloride were lower (p < or = 0.05) in the small follicles. Serum concentrations of sodium, chloride, calcium and phosphorous were higher (p < or = 0.05), while that of potassium lower (p < or = 0.05) than corresponding concentrations in the follicular fluid. The concentrations of leucine and arginine were higher (p < or = 0.05) in follicular fluid when compared with serum concentrations, while the reverse was true for other amino acids. In conclusion, this study is indicative of either low or high concentrations of certain biochemical metabolites, hormones, electrolytes and amino acids in small and large follicles for the individual roles that they play in the growth and development of follicles in the one-humped she-camel. PMID:18422863

  7. Hyperstimulation syndrome: the levels of inhibin A and B in sera and follicular fluids.

    PubMed

    Ulcova-Gallova, Zdenka; Babcova, Katka; Micanova, Zdenka; Bibkova, Katarina; Rumpik, David

    2014-04-01

    Ovarial hyperstimulation syndrome (OHSS) represents a serious problem encountered during in vitro fertilization (IVF). We examined 10 patients with OHSS and 50 women also hormonally stimulated in the process of IVF who had no complications. In all women, we evaluated the number of obtained oocytes and the level of inhibins A and B in sera and follicular fluid collected at the time of ovarial puncture, the day embryo transfer and on the day of positivity for hCG. The level of inhibin B in both fluids was significantly higher (t = 0.0403) in women with high quality of oocytes. The higher level of inhibin A was detected in patients with OHSS at the time of oocyte collection and on the day of embryo transfer. Inhibin B was elevated only at the time of oocyte collection. The levels of inhibin A and B were identical in follicular fluids collected from both ovaries. We observed no statistically significant differences between the levels of inhibin A and B in follicular fluids of women in the absence of OHSS. Evaluation of serum levels of inhibin A and B at the time of oocyte collection may contribute to the prognosis and prevention of OHSS. PMID:24455972

  8. Proteome Profile and Quantitative Proteomic Analysis of Buffalo (Bubalusbubalis) Follicular Fluid during Follicle Development.

    PubMed

    Fu, Qiang; Huang, Yulin; Wang, Zhiqiang; Chen, Fumei; Huang, Delun; Lu, Yangqing; Liang, Xianwei; Zhang, Ming

    2016-01-01

    Follicular fluid (FF) accumulates in the antrum of the ovarian follicle and provides the microenvironment for oocyte development. FF plays an important role in follicle growth and oocyte maturation. The FF provides a unique window to investigate the processes occurring during buffalo follicular development. The observed low quality of buffalo oocytes may arise from the poor follicular microenvironment. Investigating proteins found in buffalo FF (BFF) should provide insight into follicular development processes and provide further understanding of intra-follicular maturation and oocytes quality. Here, a proteomic-based approach was used to analyze the proteome of BFF. SDS-PAGE separation combined with mass spectrometry was used to generate the proteomic dataset. In total, 363 proteins were identified and classified by Gene Ontology terms. The proteins were assigned to 153 pathways, including signaling pathways. To evaluate difference in proteins expressed between BFF with different follicle size (small, <4 mm; and large, >8 mm), a quantitative proteomic analysis based on multi-dimensional liquid chromatography pre-fractionation tandem Orbitrap mass spectrometry identification was performed. Eleven differentially expressed proteins (six downregulated and five upregulated in large BFF) were identified and assigned to a variety of functional processes, including serine protease inhibition, oxidation protection and the complement cascade system. Three differentially expressed proteins, Vimentin, Peroxiredoxin-1 and SERPIND1, were verified by Western blotting, consistent with the quantitative proteomics results. Our datasets offers new information about proteins present in BFF and should facilitate the development of new biomarkers. These differentially expressed proteins illuminate the size-dependent protein changes in follicle microenvironment. PMID:27136540

  9. Proteome Profile and Quantitative Proteomic Analysis of Buffalo (Bubalusbubalis) Follicular Fluid during Follicle Development

    PubMed Central

    Fu, Qiang; Huang, Yulin; Wang, Zhiqiang; Chen, Fumei; Huang, Delun; Lu, Yangqing; Liang, Xianwei; Zhang, Ming

    2016-01-01

    Follicular fluid (FF) accumulates in the antrum of the ovarian follicle and provides the microenvironment for oocyte development. FF plays an important role in follicle growth and oocyte maturation. The FF provides a unique window to investigate the processes occurring during buffalo follicular development. The observed low quality of buffalo oocytes may arise from the poor follicular microenvironment. Investigating proteins found in buffalo FF (BFF) should provide insight into follicular development processes and provide further understanding of intra-follicular maturation and oocytes quality. Here, a proteomic-based approach was used to analyze the proteome of BFF. SDS-PAGE separation combined with mass spectrometry was used to generate the proteomic dataset. In total, 363 proteins were identified and classified by Gene Ontology terms. The proteins were assigned to 153 pathways, including signaling pathways. To evaluate difference in proteins expressed between BFF with different follicle size (small, <4 mm; and large, >8 mm), a quantitative proteomic analysis based on multi-dimensional liquid chromatography pre-fractionation tandem Orbitrap mass spectrometry identification was performed. Eleven differentially expressed proteins (six downregulated and five upregulated in large BFF) were identified and assigned to a variety of functional processes, including serine protease inhibition, oxidation protection and the complement cascade system. Three differentially expressed proteins, Vimentin, Peroxiredoxin-1 and SERPIND1, were verified by Western blotting, consistent with the quantitative proteomics results. Our datasets offers new information about proteins present in BFF and should facilitate the development of new biomarkers. These differentially expressed proteins illuminate the size-dependent protein changes in follicle microenvironment. PMID:27136540

  10. Circulating microRNAs in follicular fluid, powerful tools to explore in vitro fertilization process

    PubMed Central

    Scalici, E.; Traver, S.; Mullet, T.; Molinari, N.; Ferrières, A.; Brunet, C.; Belloc, S.; Hamamah, S.

    2016-01-01

    Circulating or “extracellular” microRNAs (miRNAs) detected in biological fluids, could be used as potential diagnostic and prognostic biomarkers of several disease, such as cancer, gynecological and pregnancy disorders. However, their contributions in female infertility and in vitro fertilization (IVF) remain unknown. This study investigated the expression profiles of five circulating miRNAs (let-7b, miR-29a, miR-30a, miR-140 and miR-320a) in human follicular fluid from 91 women with normal ovarian reserve and 30 with polycystic ovary syndrome (PCOS) and their ability to predict IVF outcomes. The combination of FF miR-30a, miR-140 and let-7b expression levels discriminated between PCOS and normal ovarian reserve with a specificity of 83.8% and a sensitivity of 70% (area under the ROC curve, AUC = 0.83 [0.73–0.92]; p < 0.0001). FF samples related to low number of mature oocytes (≤2) contained significant less miR-320a levels than those related to a number of mature oocytes >2 (p = 0.04). Moreover, FF let-7b predicted the development of expanded blastocysts with 70% sensitivity and 64.3% specificity (AUC = 0.67 [0.54–0.79]; p = 0.02) and FF miR-29a potential to predict clinical pregnancy outcome reached 0.68 [0.55–0.79] with a sensitivity of 83.3% and a specificity of 53.5% (p = 0.01). Therefore, these miRNAs could provide new helpful biomarkers to facilitate personalized medical care during IVF. PMID:27102646

  11. A Comparative Study of Serum and Follicular Fluid Leptin Concentrations among Explained Infertile, Unexplained Infertile and Fertile Women

    PubMed Central

    Kamyabi, Zahra; Gholamalizade, Tayebe

    2015-01-01

    Background The relationship between metabolism and reproduction has been always considered as an important topic in female endocrinology. It seems that leptin is one of the involved factors in infertility. Leptin, in addition to regulating body weight plays an important role in regulation of endocrine, reproductive and immune systems. The aim of this stduy is to compare serum and follicular fluid leptin concentrations in order to find the role of leptin level in infertility. Materials and Methods This case-control study was performed from September 2010 to March 2013. A total of 90 women referred to the Infertility Center of Afzalipour Hos- pital, Kerman, Iran, and divided into three equal groups (n=30/per group) of explained infertile (including 4 subgroups), unexplained infertile and normal fertile (control group). The three groups were matched in regard to demographic features [age: 20-40 years and body mass index (BMI): 20-25]. In order to determine leptin level, blood sample and fol- licular fluid were taken one hour prior and at the time of follicular puncture, respectively. Serum and follicular fluid leptin levels were measured using enzyme-linked immune sorbent assay (ELISA). Data were analyzed using descriptive-analytic tests, like Mann- Whitney and Kruskal Wallis tests, through Statistical Package for the Social Sciences (SPSS) version 16. Results In explained infertile and fertile groups, as opposed to unexplained infertile group, mean leptin level was lower in follicular fluid than in serum. Mean follicular fluid leptin concentration in women with unexplained infertility was higher com- pared to the other two groups. Women with unexplained infertility had lower level of serum leptin in comparison to the other two groups. Follicular fluid leptin level in all subgroups of explained infertile group was lower as compared to unexplained and fertile women. Conclusion The results suggested that high leptin level of follicular fluid is one of the main factors

  12. Altered Theca and Cumulus Oocyte Complex Gene Expression, Follicular Arrest and Reduced Fertility in Cows with Dominant Follicle Follicular Fluid Androgen Excess

    PubMed Central

    Summers, Adam F.; Pohlmeier, William E.; Sargent, Kevin M.; Cole, Brizett D.; Vinton, Rebecca J.; Kurz, Scott G.; McFee, Renee M.; Cushman, Robert A.; Cupp, Andrea S.; Wood, Jennifer R.

    2014-01-01

    Aspiration of bovine follicles 12–36 hours after induced corpus luteum lysis serendipitously identified two populations of cows, one with High androstenedione (A4; >40 ng/ml; mean = 102) and another with Low A4 (<20 ng/ml; mean = 9) in follicular fluid. We hypothesized that the steroid excess in follicular fluid of dominant follicles in High A4 cows would result in reduced fertility through altered follicle development and oocyte maternal RNA abundance. To test this hypothesis, estrous cycles of cows were synchronized and ovariectomy was performed 36 hours later. HPLC MS/MS analysis of follicular fluid showed increased dehydroepiandrosterone (6-fold), A4 (158-fold) and testosterone (31-fold) in the dominant follicle of High A4 cows. However, estrone (3-fold) and estradiol (2-fold) concentrations were only slightly elevated, suggesting a possible inefficiency in androgen to estrogen conversion in High A4 cows. Theca cell mRNA expression of LHCGR, GATA6, CYP11A1, and CYP17A1 was greater in High A4 cows. Furthermore, abundance of ZAR1 was decreased 10-fold in cumulus oocyte complexes from High A4 cows, whereas NLRP5 abundance tended to be 19.8-fold greater (P = 0.07). There was a tendency for reduction in stage 4 follicles in ovarian cortex samples from High A4 cows suggesting that progression to antral stages were impaired. High A4 cows tended (P<0.07) to have a 17% reduction in calving rate compared with Low A4 cows suggesting reduced fertility in the High A4 population. These data suggest that the dominant follicle environment of High A4 cows including reduced estrogen conversion and androgen excess contributes to infertility in part through altered follicular and oocyte development. PMID:25330369

  13. Origin of bovine follicular fluid and its effect during in vitro maturation on the developmental competence of bovine oocytes.

    PubMed

    Ali, Atef; Coenen, Karine; Bousquet, Daniel; Sirard, Marc-André

    2004-12-01

    Protein supplementation during in vitro maturation can profoundly affect both the rate and overall efficiency of the maturation procedure. The present study was conducted to assess the ability of different concentrations (1, 5, and 10%) of bovine follicular fluid (bFF) to support in vitro maturation of oocytes and subsequent developmental capacity. The bFF was derived either from competent follicles ( > 8 mm) obtained by transvaginal recovery following superovulation or from a pool of small follicles (2-5 mm) from abbatoir-derived ovaries. Bovine oocytes were cultured for 24 h in synthetic oviduct fluid medium (m-SOF) supplemented with polyvinylpyrrolidone. Following fertilization and embryo culture, more oocytes (P < 0.05) reached the blastocyst stage when oocytes were cultured with 5% bFF from competent follicles (41 +/- 3.7%) compared with bFF derived from small follicles (16 +/- 2.9%). Estradiol and recombinant human follicle stimulating hormone added to the competent bFF during maturation acted in synergy to increase blastocyst production rate (P < 0.05); this blastocyst production rate (57 +/- 1.2%) was higher than those obtained with the addition of these two hormones to bFF derived from small follicles (26 +/- 2.9%). The quality of blastocysts obtained was reflected by inner cell mass (51.30 +/- 3.5 and 25.50 +/- 3.7) and trophectoderm cell numbers (99.72 +/- 2.5 and 94.80 +/- 4.7) for bFF from competent and small follicles, respectively. In conclusion, follicular fluid originating from competent follicles increased the developmental competence of abbatoir-derived oocytes. PMID:15511547

  14. Divalent cation levels in serum and preovulatory follicular fluid of women undergoing in vitro fertilization embryo transfer.

    PubMed

    Azem, Foad; Hanannel, Arik; Wolf, Yoram; Ben-Yosef, Dalit; Wagman, Israel; Yovel, Israel; Kapustiansky, Rita; Lessing, Joseph B; Maclov, Mira; Amit, Ami

    2004-01-01

    In this controlled clinical study, we determined the serum and follicular fluid concentrations of the biologically active fractions of magnesium (Mg) and calcium (Ca) in 39 women undergoing controlled ovarian stimulation (COH) for in vitro fertilization (IVF) embryo transfer (ET). Serum levels of ionized Ca (Ca2+) and ionized Mg (Mg2+) were measured during day 3 of the IVF cycle, on the day of ovum pickup and 12 days following ET. Follicular fluid levels were measured on the day of ovum pickup. Serum levels of Mg2+ decreased and Ca2+ levels increased during the IVF cycle. Mg levels were significantly higher in follicular fluid than in serum. Ca2+ and Mg2+ may play a role in the preovulatory follicle, a possibility that warrants further study. PMID:14671416

  15. Characterization and Small RNA Content of Extracellular Vesicles in Follicular Fluid of Developing Bovine Antral Follicles

    PubMed Central

    Navakanitworakul, Raphatphorn; Hung, Wei-Ting; Gunewardena, Sumedha; Davis, John S.; Chotigeat, Wilaiwan; Christenson, Lane K.

    2016-01-01

    Exosomes and microvesicles (i.e., extracellular vesicles: EVs) have been identified within ovarian follicular fluid and recent evidence suggests that EVs are able to elicit profound effects on ovarian cell function. While existence of miRNA within EVs has been reported, whether EV size and concentration as well as their cargos (i.e., proteins and RNA) change during antral follicle growth remains unknown. Extracellular vesicles isolated from follicular fluid of small, medium and large bovine follicles were similar in size, while concentration of EVs decreased progressively as follicle size increased. Electron microscopy indicated a highly purified population of the lipid bilayer enclosed vesicles that were enriched in exosome biomarkers including CD81 and Alix. Small RNA sequencing identified a large number of known and novel miRNAs that changed in the EVs of different size follicles. Ingenuity Pathway Analysis (IPA) indicated that miRNA abundant in small follicle EV preparations were associated with cell proliferation pathways, while those miRNA abundant in large follicle preparations were related to inflammatory response pathways. These studies are the first to demonstrate that EVs change in their levels and makeup during antral follicle development and point to the potential for a unique vesicle-mediated cell-to-cell communication network within the ovarian follicle. PMID:27158133

  16. Can blood or follicular fluid levels of presepsin predict reproductive outcomes in ART; a preliminary study

    PubMed Central

    Ovayolu, Ali; Özdamar, Özkan; Gün, İsmet; Arslanbuga, Cansev Yılmaz; Sofuoğlu, Kenan; Tunalı, Gülden; Topuz, Samet

    2015-01-01

    Many stages of COH protocols are considered to potentiate a state of systemic inflammation. The limit beyond which inflammation has negative impacts on the formation of conception and the reproductive outcomes are compromised still remains unclear. Presepsin is a novel biomarker for diagnosing systemic inflammation and sepsis. We aimed to investigate whether plasma and follicular fluid presepsin values on oocyte pick-up (OPU) day, embryo transfer (ET) day and pregnancy test (PT) days could predict reproductive outcomes during IVF treatment in women with UEI. Patients were assigned to two groups according to pregnancy test results; pregnant (Group 1) and non-pregnant (Group 2). From all patients included in the study, 2 cc of venous blood was sampled on the three days and follicular fluid (FF) was collected during oocyte retrieval. Plasma presepsin, CRP and WBC values and FF presepsin values were measured and compared between the 2 groups. There was no significant difference between FF and plasma presepsin levels on the OPU day (298±797.4 ve 352.9±657.1; P=0.701, respectively). Plasma WBC, CRP and presepsin levels on the OPU, ET and PT days and FF presepsin levels on OPU day were not different between the 2 groups. Plasma presepsin course on the separate 3 days were different between the groups. PMID:26221358

  17. Characterization and Small RNA Content of Extracellular Vesicles in Follicular Fluid of Developing Bovine Antral Follicles.

    PubMed

    Navakanitworakul, Raphatphorn; Hung, Wei-Ting; Gunewardena, Sumedha; Davis, John S; Chotigeat, Wilaiwan; Christenson, Lane K

    2016-01-01

    Exosomes and microvesicles (i.e., extracellular vesicles: EVs) have been identified within ovarian follicular fluid and recent evidence suggests that EVs are able to elicit profound effects on ovarian cell function. While existence of miRNA within EVs has been reported, whether EV size and concentration as well as their cargos (i.e., proteins and RNA) change during antral follicle growth remains unknown. Extracellular vesicles isolated from follicular fluid of small, medium and large bovine follicles were similar in size, while concentration of EVs decreased progressively as follicle size increased. Electron microscopy indicated a highly purified population of the lipid bilayer enclosed vesicles that were enriched in exosome biomarkers including CD81 and Alix. Small RNA sequencing identified a large number of known and novel miRNAs that changed in the EVs of different size follicles. Ingenuity Pathway Analysis (IPA) indicated that miRNA abundant in small follicle EV preparations were associated with cell proliferation pathways, while those miRNA abundant in large follicle preparations were related to inflammatory response pathways. These studies are the first to demonstrate that EVs change in their levels and makeup during antral follicle development and point to the potential for a unique vesicle-mediated cell-to-cell communication network within the ovarian follicle. PMID:27158133

  18. Can blood or follicular fluid levels of presepsin predict reproductive outcomes in ART; a preliminary study.

    PubMed

    Ovayolu, Ali; Özdamar, Özkan; Gün, İsmet; Arslanbuga, Cansev Yılmaz; Sofuoğlu, Kenan; Tunalı, Gülden; Topuz, Samet

    2015-01-01

    Many stages of COH protocols are considered to potentiate a state of systemic inflammation. The limit beyond which inflammation has negative impacts on the formation of conception and the reproductive outcomes are compromised still remains unclear. Presepsin is a novel biomarker for diagnosing systemic inflammation and sepsis. We aimed to investigate whether plasma and follicular fluid presepsin values on oocyte pick-up (OPU) day, embryo transfer (ET) day and pregnancy test (PT) days could predict reproductive outcomes during IVF treatment in women with UEI. Patients were assigned to two groups according to pregnancy test results; pregnant (Group 1) and non-pregnant (Group 2). From all patients included in the study, 2 cc of venous blood was sampled on the three days and follicular fluid (FF) was collected during oocyte retrieval. Plasma presepsin, CRP and WBC values and FF presepsin values were measured and compared between the 2 groups. There was no significant difference between FF and plasma presepsin levels on the OPU day (298±797.4 ve 352.9±657.1; P=0.701, respectively). Plasma WBC, CRP and presepsin levels on the OPU, ET and PT days and FF presepsin levels on OPU day were not different between the 2 groups. Plasma presepsin course on the separate 3 days were different between the groups. PMID:26221358

  19. Metabolic profile of serum and follicular fluid from postpartum dairy cows during summer and winter.

    PubMed

    Alves, Benner G; Alves, Kele A; Martins, Muller C; Braga, Lucas S; Silva, Thiago H; Alves, Bruna G; Santos, Ricarda M; Silva, Thiago V; Viu, Marco A O; Beletti, Marcello E; Jacomini, José O; Gambarini, Maria L

    2014-01-01

    This study was designed to monitor the biochemical profiles of serum and follicular fluid (FF) of postpartum dairy cows during the summer (n=30) and winter (n=30). Blood and FF (follicles ≥ 9 mm) were obtained from Girolando cows at 30, 45, 60, 75 and 90 days postpartum. The samples were collected and analysed to determine glucose, total cholesterol (TC), triglyceride (TG), urea, sodium (Na), potassium (K) and calcium (Ca) levels. Throughout the study, the following clinical variables were measured: rectal temperature (RT), respiratory rate (RR) and body condition score (BCS). In addition, the temperature humidity index (THI) was calculated for each season. During the summer season, THI was higher, BCS decreased, there was an increase in RT, and glucose, urea, Na and K serum levels were decreased (P<0.05). The levels of TC, TG, urea, K and Ca in follicular fluid increased (P<0.05). Positive correlations (P<0.05) were observed between the serum and FF levels for glucose (r=0.29), TC (r=0.24) and Ca (r=0.30). Therefore, the biochemical profile of serum and FF of dairy cows under summer heat-stress conditions demonstrates marked changes that may impair fertility during lactation. PMID:23815864

  20. Serum but not follicular fluid cytokine levels are increased in stimulated versus natural cycle IVF: a multiplexed assay study.

    PubMed

    Bersinger, N A; Kollmann, Z; Von Wolff, M

    2014-12-01

    Throughout follicular growth the number of immune cells increases, enhanced under stimulation with exogenous gonadotropins. This treatment, however, may adversely influence folliculogenesis and negatively affect oocyte quality through modifications in the follicular concentrations of cytokines released by these immune cells. We studied this hypothesis by systematically analysing the concentrations of cytokines present in the serum and follicular fluid at the time of follicular aspiration in conventional gonadotropin-stimulated (c-IVF) cycles in comparison with natural cycle IVF (NC-IVF) in which the follicles were naturally matured. Our study involved 37 NC-IVF and 39 c-IVF cycles including 13 women who underwent both therapies. Mean age was 35.3 ± 4.6 (SD) and 34.2 ± 3.7 years in the NC-IVF and c-IVF groups (ns). Thirteen cytokines were determined in matched serum and FF samples. Interleukin (IL)-4, TNF-α, RANTES, eotaxin and interferon-gamma-induced protein-10 concentrations were lower in FF than in serum. IL-6, -8, -10, -18, monocyte chemotactic protein-1 (MCP-1), VEGF and leukaemia inhibitory factor (LIF) showed higher median levels in FF than in serum, indicating possible ovarian production. Most of these markers were also increased in concentration in the stimulated (c-IVF) than in the NC groups in the serum, but not in the follicular fluid. This finding can be attributed to the increased number of active follicles present after controlled ovarian stimulation. IL-8 was reduced in c-IVF cycles. Our study did not reveal differences in follicular fluid but in serum cytokine concentrations, suggesting that the follicular immune system might not be significantly affected by gonadotropin stimulation. PMID:25103590

  1. Effects of multivitamin/mineral supplementation on trace element levels in serum and follicular fluid of women undergoing in vitro fertilization (IVF).

    PubMed

    Özkaya, Mehmet Okan; Nazıroğlu, Mustafa; Barak, Cihan; Berkkanoglu, Murat

    2011-01-01

    We investigated effects of multivitamin/mineral supplementation on element levels in serum and follicular fluid of women undergoing IVF. We used three groups in this study. The first group was used as an age-matched and nonpregnant control (n = 13). Group 2 (n = 30) constituted the IVF group and women in the third group who were undergoing IVF also received a multivitamin/mineral tablet daily for 45 days. Follicular fluid and serum selenium and zinc levels and follicular fluid copper levels were lower in IVF patients than in controls although follicular fluid aluminum and iron levels were higher in IVF patients than in controls. However, follicular fluid and serum aluminum, copper, zinc and selenium levels, and serum magnesium levels were higher in the multivitamin/mineral group than in the IVF group although follicular fluid iron levels were lower in the multivitamin/mineral group than in the IVF group. In conclusion, we observed that copper, zinc, and selenium in serum and follicular fluid decreased in women undergoing IVF. Multivitamin/mineral supplementation in serum and follicular fluid of women undergoing IVF normalized the trace element levels. PMID:20180042

  2. Concentrations of steroids and biochemical constituents in follicular fluid of buffalo cows during different stages of the oestrous cycle.

    PubMed

    Eissa, H M

    1996-09-01

    One of the factors controlling ovarian physiology in cattle is the alteration that occurs in the hormonal and biochemical composition of follicular fluid. Because they are ill-defined in buffaloes, the present study was aimed at determining changes in the follicular fluid collected from 197 pairs of buffalo ovaries after slaughter. Total oestrogens, progesterone, calcium, inorganic phosphorus and total protein were found to be higher before oestrus and reached their peak values during oestrus. Glucose concentrations were maximal before oestrus. Testosterone, total corticosteroids and acid phosphatase concentrations increased during metoestrus peaking during dioestrus. Alkaline phosphatase concentration was higher during dioestrus. Ratios and correlations between changes in concentrations of the different follicular steroids were determined. PMID:8885467

  3. Magnesium as a regulator of thrombin formation in bovine ovarian follicular fluid.

    PubMed

    Yamada, M; Hirakushi, K; Inoue, K; Horiuchi, T; Sakai, J; Okada, T; Sugie, I

    1998-07-01

    In the ovarian follicular fluid (FF) of Holstein cows, calcium (Ca) and magnesium (Mg) levels and their roles on thrombin generation were examined and compared with the blood samples. Total Ca levels in FF increased while the total Mg levels decreased with follicular development from preantral to preovulatory stage of follicles. These changes resulted in Ca values being significantly (p < 0.05) higher in FF from the most developed follicles and the Mg values being significantly higher (p < 0.05) in the least developed follicles. To determine whether the high level of Mg might function to regulate thrombin generation in FF as occurs in plasma, the influence of Mg supplementation of FF from various types of follicles was examined. In FF from small size follicles, Mg accelerated the prothrombin time, an estimation of the overall rate of thrombin production, although a similar effects was not observed in FF from medium and large size follicles. The addition of Mg to FF from all sizes of follicles resulted an inhibition in factor X activation. Since activation of factor X is a precursor step for thrombin formation it is concluded that Mg can function as a slow accelerator of thrombin generation in FF from follicles at the antral stage of development. It is likely to have a more important role in regulating the rate of thrombin generation as the follicle develops. PMID:9713812

  4. In vitro fertilization and development of porcine oocytes matured in follicular fluid.

    PubMed

    Agung, Budiyanto; Otoi, Takeshige; Fuchimoto, Dai-ichiro; Senbon, Shoichiro; Onishi, Akira; Nagai, Takashi

    2013-01-01

    This study was conducted to assess the fertilization and development of porcine oocytes matured in a solo follicular fluid (pFF) using different in vitro culture systems and insemination periods. Cumulus-oocyte complexes (COCs), follicular cells (FCs), and pFF were collected from the follicles of ovaries. The pFF was used as a maturation medium (MpFF) after supplementation with follicle stimulating hormone (FSH) and antibiotics. The COCs were matured in a 15 ml test tube containing 3.5 ml of MpFF with FCs (5.2 × 10(6) cells/ml; rotating culture system) or 2 ml of MpFF without FCs in a 35-mm petri dish (static culture system) for 44 to 48 h. After maturation culture, oocytes were co-incubated with frozen-thawed spermatozoa for 5 h and then cultured for 7 days. The total mean rates of sperm penetration, normal fertilization, male pronucleus (MPN) formation, cleavage, and development to the blastocyst stage of oocytes after insemination were significantly higher (P<0.01) in the rotating culture system than in the static culture system. In conclusion, compared with the static culture system, the rotating culture system is adequate for the production of developmentally competent porcine oocytes when MpFF is used as a maturation medium. PMID:23428620

  5. MicroRNA Species in Follicular Fluid Associating With Polycystic Ovary Syndrome and Related Intermediary Phenotypes

    PubMed Central

    Sørensen, Anja E.; Wissing, Marie Louise; Englund, Anne Lis M.

    2016-01-01

    Context: Polycystic ovary syndrome (PCOS) has a largely unknown etiology and presents with a clinical heterogeneous patient group. Small noncoding microRNA (miRNA) might prove promising as biomarker candidates for PCOS patient stratification. Altered miRNA expression profiles have been observed in few studies. Objective: The aim was to assess the miRNA expression profile in follicular fluid from PCOS patients and healthy, regularly cycling, matched controls. Design and Setting: Experimental case-control study including 49 PCOS women (19 of which were hyperandrogenic and 30 normo-androgenic) and 21 healthy matched women all undergoing in vitro fertilization treatment. Interventions and Main Outcome: Anthropometric and relevant clinical baseline measurements were obtained. Relative expression of miRNA levels were estimated using miRNA quantitative PCR arrays and validated by quantitative RT-PCR. Correlation between miRNAs and clinical relevant measurements was estimated. Results: PCOS women, both normo-androgenic and hyperandrogenic, had decreased levels of miR-24-3p, -29a, -151-3p, and -574-3p compared with controls. Furthermore, miR-518f-3p was differentially expressed within the PCOS group with high levels observed in the hyperandrogenic group compared with the normo-androgenic PCOS patients. Serum levels of total and free T were positively correlated with miR-518f-3p in PCOS subjects (P = .001). Distinction between PCOS and controls could be made using miR-151-3p alone with an area under the curve of 0.91 or a combination of four selected miRNAs (area under the curve, 0.93). Bioinformatic target analysis points to an involvement of these miRNAs in biological pathways involving regulation of cell proliferation, extracellular matrix, and processes in intermediary metabolism. Conclusion: Our study provides evidence that the miRNA expression profile in follicular fluid is altered in PCOS and indicates that specific follicular fluid miRNAs are associated with

  6. Metabolomic changes in follicular fluid induced by soy isoflavones administered to rats from weaning until sexual maturity

    SciTech Connect

    Wang, Wenxiang; Zhang, Wenchang; Liu, Jin; Sun, Yan; Li, Yuchen; Li, Hong; Xiao, Shihua; Shen, Xiaohua

    2013-06-15

    Female Wistar rats at 21 days of age were treated with one of three concentrations of soy isoflavones (SIF) (50, 100 or 200 mg/kg body weight, orally, once per day) from weaning until sexual maturity (3 months) in order to evaluate the influence of SIF on ovarian follicle development. After treatment, the serum sex hormone levels and enumeration of ovarian follicles of the ovary were measured. The metabolic profile of follicular fluid was determined using HPLC-MS. Principal component analysis (PCA) and partial least-squares-discriminant analysis (PLS-DA) was used to identify differences in metabolites and reveal useful toxic biomarkers. The results indicated that modest doses of SIF affect ovarian follicle development, as demonstrated by decreased serum estradiol levels and increases in both ovarian follicle atresia and corpora lutea number in the ovary. SIF treatment-related metabolic alterations in follicular fluid were also found in the PCA and PLS-DA models. The 24 most significantly altered metabolites were identified, including primary sex hormones, amino acids, fatty acids and metabolites involved in energy metabolism. These findings may indicate that soy isoflavones affect ovarian follicle development by inducing metabolomic variations in the follicular fluid. - Highlights: ► Modest doses of soy isoflavones (SIF) do affect ovarian follicle development. ► SIF treatment-related metabolic alterations in follicular fluid were found. ► The 24 most significantly altered metabolites were identified.

  7. Analysis of follicular fluid total phospholipids in women undergoing in-vitro fertilisation.

    PubMed

    Fayezi, S; Darabi, M; Darabi, M; Nouri, M; Rahimipour, A; Mehdizadeh, A

    2014-04-01

    Follicular fluid (FF) samples were obtained from 100 patients referred to the University Hospital. A total of 79 subjects underwent IVF and the remaining 21 underwent ICSI. The levels of apoA-I and total phospholipid were measured using turbidometric and colorimetric phosphorus assays, respectively. Correlation analysis showed a significant inverse association of total phospholipid in FF with fertilisation ratio (r = -0.24, p = 0.04). Furthermore, the ratio of phospholipid/apoA-I in patients with a percentage of fertilised oocytes ≤ 50% was significantly higher (> 2.5%, p < 0.05) than in those with higher percentages of fertilised oocytes. The amounts of phospholipid and phospholipid/apoA-I ratio in FF were associated negatively to the percentage of oocyte fertilisation. Therefore, the change in the phospholipid and phospholipid/apoA-I ratio of FF might be regarded as indicators of female fertility. PMID:24476398

  8. Granulosa cell apoptosis, aromatase activity, cyclic adenosine 3',5'-monophosphate response to gonadotropins, and follicular fluid steroid levels during spontaneous and induced follicular atresia in ewes.

    PubMed

    Jolly, P D; Tisdall, D J; De'ath, G; Heath, D A; Lun, S; Hudson, N L; McNatty, K P

    1997-04-01

    The aims of the present study in ewes were 1) to test the hypothesis that apoptosis in granulosa cells is one of the processes involved in the structural demise of follicles and 2) to define the temporal relationships among the occurrence and degree of apoptosis in granulosa cells, aromatase activity, production of cyclic AMP (cAMP) by granulosa cells in response to FSH or LH, concentrations of estradiol 17 beta (E2) and progesterone in follicular fluid, and the characteristic morphometric changes associated with the process of follicular atresia. To address these aims, ewes were treated with either saline or steroid-free bovine follicular fluid (bFF) at 60 h after estrus, and ovarian follicles > or = 3 mm diameter were recovered at 0, 12, 18, or 24 h later. Apoptotic granulosa cells were identified by the presence of oligonucleosomes after 3'-end labeling of extracted DNA with [32P]alpha dideoxy ATP (ddATP). The degree of oligonucleosome formation, based on the intensity of radiolabeling, was given an apoptosis score (AP) of 0 (nondetectable), 1 (slight), 2 (moderate), or 3 (marked). Moreover, a labeling index (LI) was calculated from the amount of radiolabeled ddATP incorporated into low-molecular weight (< 4.2 kb) DNA fragments. On the basis of morphometric criteria, 73% (141 of 194) of the follicles classified as healthy had apoptotic granulosa cells compared to 86% (18 of 21) of the follicles classified as atretic. In the bFF-but not saline-treated ewes, the concentrations of plasma FSH had declined to basal values at 12 h after treatment. At the beginning of the treatment period, the degree of granulosa cell apoptosis was either undetectable (AP = 0, 47% of follicles) or slight (AP = 1, 44% of follicles) in the majority of follicles. After 12 h from the bFF but not the saline injection, there was a significant increase in the proportion of follicles (> or = 3 mm diameter) per ewe containing apoptotic granulosa cells (p < 0.001) and a significant decrease in

  9. Insulin-like growth factor binding proteins in follicular fluid from morphologically distinct healthy and atretic bovine antral follicles.

    PubMed

    Irving-Rodgers, H F; Catanzariti, K D; Master, M; Grant, P A; Owens, P C; Rodgers, R J

    2003-01-01

    In bovine follicles 2-5 mm in diameter, two morphologically distinct types of healthy follicles and two types of atretic follicles have been described recently. Healthy follicles either have columnar basal granulosa cells with follicular basal lamina composed of many layers or 'loops' or they have rounded basal cells with a conventional single-layered, aligned follicular basal lamina. In atretic follicles, cell death either commences at the basal layer and progresses to the antrum (basal atresia) with macrophage penetration of the membrana granulosa or death progresses from the antrum in a basal direction (antral atresia). Little is known about how these different phenotypes develop. To determine whether insulin-like growth factor binding protein (IGFBP) levels in follicular fluid differ between these different types of follicles, we measured IGFBP levels in fluids from these follicles. A total of 61 follicles were assessed by light microscopy and characterized by morphological analysis as either healthy, with columnar or rounded basal granulosa cells, or as undergoing antral or basal atresia. The IGFBP concentration in the follicular fluid of individual follicles from the four groups (n = 12-20 per group) was identified by Western ligand blots using (125)I-insulin-like growth factor (IGF)-II as a probe. Insulin-like growth factor binding proteins 2, 3 (44 and 40 kDa), 4 (glycosylated and non-glycosylated) and 5 were observed. The levels (per volume of fluid) of IGFBPs 2, 4 and 5 were greater in atretic follicles than in healthy follicles. However, there were no statistical differences in levels of each IGFBP between either the two types of healthy follicle or between the two types of atretic follicles. Thus, IGFBP levels are not related to the different types of healthy or atretic follicles. PMID:12921699

  10. Follicular fluid norepinephrine and dopamine concentrations are higher in polycystic ovary syndrome.

    PubMed

    Musalı, Natı; Özmen, Batuhan; Şükür, Yavuz Emre; Ergüder, Berrin İmge; Atabekoğlu, Cem Somer; Sönmezer, Murat; Berker, Bülent; Aytaç, Ruşen

    2016-06-01

    The aim of the present study was to compare follicular fluid (FF) levels of norepinephrine (NE) and dopamine (DA) in polycystic ovary syndrome (PCOS) and non-PCOS patients who underwent in vitro fertilization (IVF). Forty-seven PCOS patients (study group) and 61 patients with male factor infertility (control group) who underwent IVF using GnRH agonist protocol were recruited. Concentrations of NE and DA were measured in FF specimens of all patients. Demographic characteristics were comparable between the groups. Significantly higher levels of NE were measured in FF of PCOS patients (median: 61.05 nmol/l) compared to those with male infertility (median: 49.82 nmol/l). Similarly, significantly higher levels of DA were measured in FF of PCOS patients (median: 23.70 nmol/l) compared to those with male infertility (median: 18.28 nmol/l). In conclusion, the FF concentrations of both catecholamine are increased in PCOS patients when compared to non-PCOS patients. PMID:26754116

  11. Comparison between the characteristics of follicular fluid and the developmental competence of bovine oocytes.

    PubMed

    Iwata, H; Inoue, J; Kimura, K; Kuge, T; Kuwayama, T; Monji, Y

    2006-02-01

    There are great differences in the developmental competence of oocytes collected from individual cows. Oocytes grow and mature in the follicular fluid (FF). In the present study, characteristics of the FF of each ovary and the developmental competence of enclosed oocytes were investigated, and these data were then compared. A total of 37 pairs of ovaries were collected from beef heifers. The concentration of magnesium (Mg), aspirate aminotransferase (AST), nonesterified fatty acids (NEFA), and lactate dehydrogenase (LDH) in the FF were great compared with serum standard. Several significant correlations among these characteristics were detected. Forty-eight hours after fertilization, the stage of embryo development at an advanced developmental stage (>6 cell stage) is related to the rate of blastulation 8 days after fertilization. In addition, a significantly positive or negative correlation was observed between the developmental competence (the rate of cleavage in the embryo and blastulation) and the concentration of the icterus index (ICT) or blood urea nitrogen (BUN) in the FF. In conclusion, the quality of oocytes is affected by the environment in the follicle, and BUN or ICT is a predictable index of the developmental competence of oocytes. PMID:15961265

  12. Purification and characterization of lutropin receptor from membranes of pig follicular fluid

    SciTech Connect

    Yarney, T.A.; Sairam, M.R.; Bhargavi, G.N.; Mohapatra, S.K. )

    1990-04-10

    Membranes derived from free floating granulosa cells in porcine ovarian follicular fluid were used as a starting material for structural characterization of both LH/hCG and FSH receptors. The receptors were highly hormone-specific and showed single classes of high-affinity binding sites. Their molecular weights as determined by affinity cross-linking with their respective {sup 125}I-ligands were similarly 70,000. The membrane-localized receptors could be solubilized with reduced Triton X-100 in the presence of 20% glycerol with good retention of hormone binding activity. The purified receptor exhibited a high specificity for hCG and hLH but not for hFSH bTSH. The purified receptor was iodinated and visualized to be composed of a major protein of M{sub r} 70,000 and other minor proteins of molecular weights ranging from 14,000 to 40,000. Except for the M{sub r} 14,000 protein, all other protein species bound to the concanavalin A-Sepharose column. The data suggest that the ovarian LH/hCG and FSH receptors are structurally similar and consist of a single polypeptide chain, as recently documented for the LH/hCG receptor.

  13. N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages.

    PubMed

    Tharmalingam-Jaikaran, T; Walsh, S W; McGettigan, P A; Potter, O; Struwe, W B; Evans, A C O; Rudd, P M; Carrington, S D

    2014-12-01

    Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. PMID:25212784

  14. Follicular fluid and urinary concentrations of phthalate metabolites among infertile women and associations with in vitro fertilization parameters.

    PubMed

    Du, Yao-Yao; Fang, Yue-Li; Wang, Yi-Xin; Zeng, Qiang; Guo, Na; Zhao, Hua; Li, Yu-Feng

    2016-06-01

    Evidence from toxicological studies has demonstrated that phthalates can lead to reduced fertility through effects on folliculogenesis, oocyte maturation and embryonic development, but human data are limited. Concentrations of eight phthalate metabolites in 110 follicular fluid (FF) and urine samples collected from 112 women attending an infertility clinic in Wuhan, China were quantified, and correlations between paired matrices were explored. Associations between metabolite concentrations and in vitro fertilization (IVF) parameters were evaluated with multivariable models. Six metabolites were detected in >72.73% of the FF samples. MEHP and MBP were the dominant metabolites with a median level of 2.80 and 2.05ng/mL, respectively. Significant correlations between the two matrices, urine and FF, were found for MEP (rs=0.44), and MBP (rs=0.22). FF and urinary metabolite concentrations were not associated with any IVF parameters. However, given the prevalence of phthalates exposure, further work is needed to elucidate the potential hazard on female reproduction. PMID:27067915

  15. Glutathione S-transferase activity in follicular fluid from women undergoing ovarian stimulation: role in maturation.

    PubMed

    Meijide, Susana; Hernández, M Luisa; Navarro, Rosaura; Larreategui, Zaloa; Ferrando, Marcos; Ruiz-Sanz, José Ignacio; Ruiz-Larrea, M Begoña

    2014-10-01

    Female infertility involves an emotional impact for the woman, often leading to a state of anxiety and low self-esteem. The assisted reproduction techniques (ART) are used to overcome the problem of infertility. In a first step of the in vitro fertilization therapy women are subjected to an ovarian stimulation protocol to obtain mature oocytes, which will result in competent oocytes necessary for fertilization to occur. Ovarian stimulation, however, subjects the women to a high physical and psychological stress, thus being essential to improve ART and to find biomarkers of dysfunction and fertility. GSH is an important antioxidant, and is also used in detoxification reactions, catalysed by glutathione S-transferases (GST). In the present work, we have investigated the involvement of GST in follicular maturation. Patients with fertility problems and oocyte donors were recruited for the study. From each woman follicles at two stages of maturation were extracted at the preovulatory stage. Follicular fluid was separated from the oocyte by centrifugation and used as the enzyme source. GST activity was determined based on its conjugation with 3,4-dichloronitrobenzene and the assay was adapted to a 96-well microplate reader. The absorbance was represented against the incubation time and the curves were adjusted to linearity (R(2)>0.990). Results showed that in both donors and patients GST activity was significantly lower in mature oocytes compared to small ones. These results suggest that GST may play a role in the follicle maturation by detoxifying xenobiotics, thus contributing to the normal development of the oocyte. Supported by FIS/FEDER (PI11/02559), Gobierno Vasco (Dep. Educación, Universiades e Investigación, IT687-13), and UPV/EHU (CLUMBER UFI11/20 and PES13/58). The work was approved by the Ethics Committee of the UPV/EHU (CEISH/96/2011/RUIZLARREA), and performed according to the UPV/EHU and IVI-Bilbao agreement (Ref. 2012/01). PMID:26461371

  16. Stromal cell contribution to human follicular lymphoma pathogenesis.

    PubMed

    Mourcin, Frédéric; Pangault, Céline; Amin-Ali, Rada; Amé-Thomas, Patricia; Tarte, Karin

    2012-01-01

    Follicular lymphoma (FL) is the prototypical model of indolent B cell lymphoma displaying a strong dependence on a specialized cell microenvironment mimicking normal germinal center. Within malignant cell niches in invaded lymph nodes and bone marrow, external stimuli provided by infiltrating stromal cells make a pivotal contribution to disease development, progression, and drug resistance. The crosstalk between FL B cells and stromal cells is bidirectional, causing activation of both partners. In agreement, FL stromal cells exhibit specific phenotypic, transcriptomic, and functional properties. This review highlights the critical pathways involved in the direct tumor-promoting activity of stromal cells but also their role in the organization of FL cell niche through the recruitment of accessory immune cells and their polarization to a B cell supportive phenotype. Finally, deciphering the interplay between stromal cells and FL cells provides potential new therapeutic targets with the aim to mobilize malignant cells outside their protective microenvironment and increase their sensitivity to conventional treatment. PMID:22973275

  17. Effect of domperidone supplementation of fescue-fed heifers on plasma and follicular fluid fatty acid composition and oocyte quality.

    PubMed

    Jones, K L; King, S S

    2009-07-01

    This study continues a series of investigations evaluating fescue endotoxin exposure in beef heifer production. The objectives were to evaluate fatty acid compositions in plasma and follicular fluid, and to assess oocyte quality from cattle fed fescue diets. The ability of domperidone, a dopamine antagonist, to mitigate these variables was also assessed. Thirty heifers were divided into 3 treatment groups (n = 10/group) and administered treatment regimens for 24 d, at which time blood samples were collected. The treatment regimens were a diet with endophyte-free fescue (EF), a diet with endophyte-infected fescue (EI), or EI supplemented with daily subcutaneous injections of domperidone (0.44 mg/kg of BW; EID). Three heifers/group were administered treatments for an additional 10 d, at which time their luteal phase ovarian follicular fluid and oocytes were collected. Plasma and follicular fluid samples were analyzed to determine fatty acid concentrations. Oocytes were matured in vitro to assess quality. In addition, abattoir oocytes were cultured in plasma from treated heifers. In plasma, arachidonic acid was less (P < 0.001) in EF-fed compared with EI-fed heifers. Decreased (P < 0.05) total n-6 fatty acid concentration was observed in EF-fed compared with EI-fed heifers. Similarly, the EF-containing diets decreased (P < 0.05) concentrations of eicosapentaenoic acid and C22:5n-3 (P < 0.05) compared with EI-containing diets. Domperidone supplementation increased (P < 0.05) C18:2 cis-9, trans-11, C17:1n-7, and several C18:1 isomers compared with the diet with EI and no supplementation. No differences between fescue endophyte groups were detected in any of the fatty acid concentrations analyzed in follicular fluid from small follicles. In follicular fluid from large follicles, C18:4n-3 and C22:6n-3 concentrations were greater (P < 0.05) in EI-fed compared with EF-fed heifers. Oocytes cultured in serum (control) or plasma from EF-, EI-, or EID-fed cattle did not differ

  18. Characterization of adiponectin concentrations and molecular weight forms in serum, seminal plasma, and ovarian follicular fluid from cattle.

    PubMed

    Heinz, Johanna F L; Singh, Shiva P; Janowitz, Ulrich; Hoelker, Michael; Tesfaye, Dawit; Schellander, Karl; Sauerwein, Helga

    2015-02-01

    Adiponectin (AdipoQ), an adipocyte-derived hormone, is one of the most abundant adipokines in the blood circulation. Adiponectin has various metabolic functions, such as improving insulin sensitivity in humans and rodents. The role of AdipoQ in reproduction is not yet fully understood, but the expression of AdipoQ in reproductive tissues has been observed in various animals and humans, including chicken testis, bovine ovary, and human placenta. The objective of this study was to characterize AdipoQ in the bovine body fluids related to reproduction. Therefore, we evaluated the seminal plasma (SP) from breeding bulls (n = 29) and follicular fluid (FF) from heifers (n = 14), and we also collected blood samples from these animals. In addition, blood samples from other bulls (n = 30) and heifers (n = 14) were assayed for AdipoQ. The concentrations were assessed using a bovine-specific ELISA, and the molecular weight (MW) pattern of the AdipoQ protein was estimated by the Western blot analysis. The SP AdipoQ concentrations were approximately 180-fold lower compared with that in the serum concentrations. Furthermore, the AdipoQ concentrations in the serum and SP were positively correlated. The MW patterns of AdipoQ in the serum and SP were different such that the high MW form of AdipoQ was more abundant in the SP than serum. The AdipoQ concentrations in the serum and SP also increased with age: old bulls (>6 years) had higher AdipoQ concentrations in the serum and SP than bulls aged 24 months or lesser (P < 0.05). In the FF, the AdipoQ concentrations were 1.6-fold lower than those in the corresponding serum samples, and the concentrations in the serum and FF were not correlated (P > 0.1). In the FF, only the middle MW forms of AdipoQ were detectable by Western blotting. The MW pattern in the serum did not differ between the sexes. Our data provide both the AdipoQ concentration and the MW patterns for bovine body fluids related to reproduction. PMID:25468552

  19. Association between Follicular Fluid Leptin and Serum Insulin Levels in Nonoverweight Women with Polycystic Ovary Syndrome

    PubMed Central

    Garruti, G.; de Palo, R.; Rotelli, M. T.; Nocera, S.; Totaro, I.; Nardelli, C.; Panzarino, M. A.; Vacca, M.; Selvaggi, L. E.; Giorgino, F.

    2014-01-01

    Aims. We evaluated the links between leptin and visfatin levels and fertilization rates in nonoverweight (NOW) women with PCOS (NOW-PCOS) from Apulia undergoing in vitro fertilization/embryo transfer (IVF). Materials and Methodology. We recruited 16 NOW women with PCOS (NOW-PCOS) and 10 normally ovulating NOW women (control-NOW). All women underwent IVF. Androgens, 17-β-estradiol (17β-E2), and insulin levels were measured in plasma and/or serum and leptin and visfatin levels were assayed in both serum and follicular fluid (FF-leptin, FF-visfatin). Results. In NOW-PCOS, both serum and FF-leptin were significantly lower than in control-NOW. In NOW-PCOS, significant correlations were found between BMI and serum leptin and insulinemia and FF-leptin. By contrast, in control-NOW, FF-leptin levels were not correlated with insulinemia. Serum visfatin levels were not significantly different in NOW-PCOS and control-NOW, but FF-visfatin levels were 1.6-fold higher, although not significantly, in NOW-PCOS than in control-NOW. Conclusions. Both serum leptin levels and FF-leptin are BMI- and insulin-related in Southern Italian NOW-PCOS from Apulia. In line with other reports showing that FF-leptin levels are predictive of fertilization rates, lower than normal FF-leptin levels in NOW-PCOS may explain their lower fertilization rate and this may be related to the level of insulin and/or insulin resistance. PMID:24895638

  20. Lower growth factor expression in follicular fluid undergone in-vitro fertilization

    PubMed Central

    Park, Seung Bin; Park, Bang Ja

    2011-01-01

    Objective This study was performed to identify whether growth and differentiation factor-9 (GDF-9) and transforming growth factor-β1 (TGF-β1) expressions would be lower in the follicular fluid (FF) of those over age 35 who underwent IVF than under age 35. Methods A total of 24 IVF cycles (20 patients) were included in this study. All of patients were stimulated for IVF by the GnRH short protocol and divided into two groups for analysis, according to their age: <35 group (14 cycles, 11 patients) vs. ≥35 group (10 cycles, 9 patients). The expression levels of GDF-9 and TGF-β1 were determined by western blotting and quantitative enzyme-linked immunosorbent assay. Results The numbers of retrieved oocytes and metaphase II oocytes were significantly lower in the ≥35 group. Lower expression of GDF-9 and TGF-β1 by western blotting in the ≥35 group were observed as well. The mean GDF-9 and TGF-β1 levels by enzyme-linked immunosorbent assay were lower in the ≥35 group. The values were 6,850.5±928.4 ng/L vs. 3,333.3±1,089.2 ng/L of GDF-9 (p<0.05) and 3,844.1±571.1 ng/L vs. 2,187.7±754.0 ng/L of TGF-β1 (p<0.05). A negative correlation between GDF-9 and age was observed (r=-0.546, p=0.006). Conclusion GDF-9 and TGF-β1 production from stimulated ovaries during IVF appears to decrease with age. PMID:22384444

  1. Liver condition affects bovine oocyte qualities by changing the characteristics of follicular fluid and plasma.

    PubMed

    Tanaka, H; Shibano, K; Monji, Y; Kuwayama, T; Iwata, H

    2013-08-01

    The liver is an important organ that contributes to milk production in dairy cows. The aim of this study was to examine whether liver conditions affect the characteristics of blood plasma and follicular fluid (FF) and whether supplementing in vitro maturation medium with FF from either cows with damaged livers (DL) or those with healthy livers (HL) affects oocyte developmental competence. Biochemical characteristics of FF were significantly correlated with those in plasma. As such, the characteristics of both plasma and FF were similarly affected by liver conditions in that the concentrations of total protein and inorganic phosphorus were higher for the DL cow group than for the HL cow group, whereas the concentrations of albumin, lactate dehydrogenase and calcium were lower for DL cows than for HL cows. In addition, supplementing the medium with DL-FF retarded the progression of the nuclear maturation of oocytes collected from the HL cows. On culturing oocytes in maturation medium containing HL-FF, DL-FF or foetal calf serum, the highest developmental rate to the blastocyst stage was observed in the HL-FF group, while the lowest developmental ratio was observed in the DL-FF group. The growth factor array of the FFs revealed that 10 growth factors were significantly downregulated in the DL-FF compared with those in HL-FF. In conclusion, the characteristics of plasma and FF are affected by liver conditions in a similar way. Concentrations of several growth factors were low in DL-FF, as was the ability of DL-FF to support oocyte maturation compared with that of HL-FF. PMID:23281835

  2. Expression of CD11c+HLA-DR+dendritic cells and related cytokines in the follicular fluid might be related to pathogenesis of ovarian hyperstimulation syndrome

    PubMed Central

    Shi, Sen-Lin; Peng, Zhao-Feng; Yao, Gui-Dong; Jin, Hai-Xia; Song, Wen-Yan; Yang, Hong-Yi; Xue, Ru-Yue; Sun, Ying-Pu

    2015-01-01

    Objective: To explore the expressions of CD11c+HLA-DR+dentritic cells in the follicular fluid of patients with OHSS and their significances. Subjects: 100 individuals. Treatment: embryos were observed. The distribution of dentritic cells in follicular fluid and the levels of IL-10, IL-12, IL-18 and IL-23 in follicular fluid were detected. Methods: There were ovarian hyperstimulation syndrome (OHSS) group and control group in this study. The OHSS group consisted of 50 patients with OHSS and the control group consisted of 50 patients who underwent in vitro fertilization-embryo transfer (IVF-ET) only due to male factors. The statuses of embryos were compared between the two groups. The distribution of dentritic cells in follicular fluid was determined with flow cytometry, and the levels of IL-10, IL-12, IL-18 and IL-23 in follicular fluid were detected with enzyme-linked immunosorbent assay (ELISA) in all patients. Results: The two-pronuclear (2PN) fertility rate, high-quality embryo rate and available embryo rate were all significantly lower in OHSS group than in control group (all P<0.05). The number of CD11c+HLA-DR+dentritic cells (P<0.05) and the levels of IL-10, IL-12, IL-18 and IL-23 were all significantly higher in OHSS group than in control group (all P<0.01). Conclusion: The follicular fluid of the patients with OHSS is in an inflammatory status, the inflammatory status may be involved in OHSS and the microenvironment of follicular fluid may affects oocyte quality and embryo development. PMID:26823856

  3. Ovarian activity and oocyte quality associated with the biochemical profile of serum and follicular fluid from Girolando dairy cows postpartum.

    PubMed

    Alves, Benner G; Alves, Kele A; Lúcio, Aline C; Martins, Muller C; Silva, Thiago H; Alves, Bruna G; Braga, Lucas S; Silva, Thiago V; Viu, Marco A O; Beletti, Marcelo E; Jacomini, José O; Santos, Ricarda M; Gambarini, Maria L

    2014-05-01

    This study was designed to evaluate the influence of heat stress (HS) on the metabolic profile of serum and follicular fluid (FF), ovarian follicle development, and oocyte quality of Girolando dairy cows. Oocytes, blood, and FF (follicles ≥9mm) samples were obtained at 30, 45, 60, 75, and 90 days postpartum in the summer and winter seasons. During transvaginal follicular aspiration, rectal temperature (RT), body condition score (BCS), number of ovarian follicles, and quality of oocytes were recorded. The ambient air temperature (AT) and relative humidity (RH) were also recorded to calculate the temperature humidity index (THI). Glucose, total cholesterol (TC), triglycerides (TG), urea, sodium (Na), potassium (K), and calcium (Ca) concentrations were determined using serum and FF samples. The RT, THI, and BCS loss were greater (P<0.01) in the summer; however, glucose, Na, and K serum concentrations decreased in the same season (P<0.05). Degenerated oocytes were positively associated (P<0.05) with THI (r=0.14) and AT (r=0.13), and negatively associated with glucose (r=-0.12) and K (r=-0.11) serum concentrations. HS induces metabolic changes, which compromise the number of ovarian follicles and the follicular environment, thus resulting in morphologically damaged oocytes. PMID:24674823

  4. Granulosa cell gene expression profiling in cows with divergent follicular fluid concentrations of androgens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inefficiencies in reproduction result in significant production costs for beef producers in the United States. In the UNL cow-calf herd, two sub-populations of cows have been identified with divergent levels of follicular androstenedione (A4). Furthermore, High A4 cows (A4 > 40ng/ml) have a 10 perce...

  5. Oocyte quality reflected by follicular fluid analysis in poly cystic ovary syndrome (PCOS): a hypothesis based on intermediates of energy metabolism.

    PubMed

    Arya, Bikas K; Haq, Ahtesham Ul; Chaudhury, Koel

    2012-04-01

    There is no clear acceptance of specific follicular fluid biomarker and its correlation with oocyte quality or related embryo variable till now. Most of the studies analyze correlation between certain biomolecules and the oocyte quality using single variable, instead of multivariate analysis algorithms. Our hypothesis is not based on single biomarker discovery, but attempts to explain oocyte quality in terms of energy metabolic pathways by considering its various intermediates. Reduced availability of glucose in the oocytes and follicular cells caused by defective transportation of glucose is expected in polycystic ovary syndrome (PCOS). This initiates alternative pathways to utilize fatty acid, amino acids etc. for energy as a compensatory mechanism to deal with the energy requirement. These compensations can be reflected by altered levels of various biomolecules in follicular fluid (e.g. ketone bodies, lipids, amino acids, lactate, pyruvate etc.). The amount of compensation, in order to meet the energy requirement can be directly correlated to quality of oocytes and better outcome after in vitro fertilization (IVF) in PCOS cases. This can be predicted with fair accuracy by doing a multivariate analysis of altered levels of various biomolecules in follicular fluid. Various supervised and unsupervised classification techniques based on spectroscopic data, obtained from follicular fluid samples may certainly prove to be an important tool to predict oocytes quality and IVF outcome with better accuracy in women with PCOS. PMID:22336088

  6. Nuclear magnetic resonance spectroscopy of bovine ovarian follicular fluid at four selected times of the oestrous cycle

    PubMed Central

    Sarty, Gordon E.; Kendall, Edward J.; Adams, Gregg P.; Pierson, Roger A.

    2010-01-01

    The objective of the study was to determine if nuclear magnetic resonance (NMR) spectral features of ovarian follicular fluid were correlated with the physiological status of follicles so that we could assess the feasibility of using NMR spectroscopy during assisted reproduction therapy. Thirty-five sexually mature, nullparious heifers were monitored by transrectal ultrasonography to assess their follicle wave status during the oestrous cycle. Ovariectomies were performed on Day 3 of wave 1 (D3W1, n = 10), Day 6 of wave 1 (D6W1, n = 9), Day 1 of wave 2 (D1W2, n = 9), or in the immediate preovulatory period of at least 17 days after ovulation (D ≥ 17, n = 9). Follicle status was determined to be dominant or subordinate. Follicular fluid was extracted from the follicles and NMR spectra were collected. Principal components were extracted from ratios of line amplitudes and tested for effects of follicle status (dominant v. subordinate) and cycle time point (D1W3, D1W6, D1W2 and D ≥ 17) using multivariate analysis of variance. For most line ratio combinations, main effects of status, time point and their interaction were found (P < 0.05). We concluded that NMR spectra may be used for the determination of ovarian follicle physiological status. PMID:16836963

  7. Amino acid and fatty acid composition of follicular fluid as predictors of in-vitro embryo development.

    PubMed

    Sinclair, K D; Lunn, L A; Kwong, W Y; Wonnacott, K; Linforth, R S T; Craigon, J

    2008-06-01

    The value of using the amino acid and fatty acid composition of follicular fluid as predictors of embryo development was assessed in a bovine model of in-vitro maturation (IVM), IVF and blastocyst culture (IVC). A total of 445 cumulus-oocyte complexes (COC) aspirated from visually healthy follicles underwent IVM and IVF singly (n = 138) or in groups (n = 307). Of these COC, 349 cleaved (78%) following IVF and 112 went on to form blastocysts (32% of cleaved) following IVC. Culture method (singly or in groups) had no effect on development. In contrast to fatty acids, which had no predictive value, the amino acid composition of follicular fluid was associated with morphological assessments of COC quality and with post-fertilization development to the blastocyst stage. Principal component analysis identified two amino acids (i.e. alanine and glycine) that had the highest value for predicting early post-fertilization development. The predictive value of these two amino acids, in terms of the percentage of oocytes that cleaved following IVF, was greatest for COC with the poorest morphological grades but, with respect to blastocyst yields, was independent of morphological grade, and so may serve as a useful additional non-invasive measure of COC quality. PMID:18549697

  8. Metabolite and ionic composition of follicular fluid from different-sized follicles and their relationship to serum concentrations in dairy cows.

    PubMed

    Leroy, J L M R; Vanholder, T; Delanghe, J R; Opsomer, G; Van Soom, A; Bols, P E J; de Kruif, A

    2004-02-01

    Metabolic changes in blood serum may be reflected in the biochemical composition of follicular fluid and could indirectly influence oocyte quality. The purpose of this study was to examine the biochemical composition of follicular fluid harvested from different-sized follicles and its relationship with that of blood serum in dairy cattle. Following slaughter, blood samples were collected from dairy cows n=30 and follicular fluid aspirated from three size classes of non-atretic follicles (<4 mm, 6-8 mm and >10 mm diameter). Samples remained independent between cows and between size classes within cows. Serum and follicular fluid samples were assayed using commercial clinical and photometric chemistry assays for ions (sodium, potassium and chloride) and metabolites (glucose, beta-hydroxybutyrate (beta-OHB), lactate, urea, total protein, triglycerides, non-esterified fatty acids (NEFA) and total cholesterol). Results showed that follicular fluid concentrations of glucose, beta-OHB and total cholesterol increased from small to large follicles and decreased for potassium, chloride, lactate, urea and triglycerides. There was a significant concentration gradient for all variables between their levels in serum and follicular fluid (P<0.05). Significant correlations were observed for chloride (r=0.40), glucose (r=0.56), beta-OHB (r=0.85), urea (r=0.95) and total protein (r=0.60) for all three follicle size classes and for triglycerides (r=0.43), NEFA (r=0.50) and total cholesterol (r=0.42) for large follicles (P<0.05). The results from the present study suggest that the oocyte and the granulosa cells of dairy cows grow and mature in a biochemical environment that changes from small to large follicles. Furthermore, the significant correlation between the composition of serum and follicular fluid for the above-mentioned metabolites suggests that metabolic changes in serum levels will be reflected in the follicular fluid and, therefore, may affect the quality of both the

  9. Proteome dataset of pre-ovulatory follicular fluids from less fertile dairy cows.

    PubMed

    Zachut, Maya; Sood, Pankaj; Livshitz, Lilya; Kra, Gitit; Levin, Yishai; Moallem, Uzi

    2016-06-01

    This article contains raw and processed data related to research published in Zachut et al. (2016) [1]. Proteomics data from preovulatory follicles in cows was obtained by liquid chromatography-mass spectrometry following protein extraction. Differential expression between controls and less fertile cows (LFC) was quantified using MS1 intensity based label-free. The only previous proteomic analysis of bovine FF detected merely 40 proteins in follicular cysts obtained from the slaughterhouse (Maniwa et al., 2005) [2], and the abundance of proteins in the bovine preovulatory FF remains unknown. Therefore, the objectives were to establish the first dataset of FF proteome in preovulatory follicles of cows, and to examine differentially expressed proteins in FF obtained in-vivo from preovulatory follicles of less fertile cows (also termed "repeat breeder") and control (CTL) cows. The proteome of FF from 10 preovulatory follicles that were aspirated in vivo (estradiol/progesterone>1) was analyzed. This novel dataset contains 219 identified and quantified proteins in FF, consisting mainly of binding proteins, proteases, receptor ligands, enzymes and transporters. In addition, differential abundance of 8 proteins relevant to follicular function was found in LFC compared to CTL; these findings are discussed in our recent research article Zachut et al. (2016) [1]. The present dataset of bovine FF proteome can be used as a reference for any study involving disorders of follicular development in dairy cows or in comparative studies between species. PMID:27182550

  10. Proteome dataset of pre-ovulatory follicular fluids from less fertile dairy cows

    PubMed Central

    Zachut, Maya; Sood, Pankaj; Livshitz, Lilya; Kra, Gitit; Levin, Yishai; Moallem, Uzi

    2016-01-01

    This article contains raw and processed data related to research published in Zachut et al. (2016) [1]. Proteomics data from preovulatory follicles in cows was obtained by liquid chromatography-mass spectrometry following protein extraction. Differential expression between controls and less fertile cows (LFC) was quantified using MS1 intensity based label-free. The only previous proteomic analysis of bovine FF detected merely 40 proteins in follicular cysts obtained from the slaughterhouse (Maniwa et al., 2005) [2], and the abundance of proteins in the bovine preovulatory FF remains unknown. Therefore, the objectives were to establish the first dataset of FF proteome in preovulatory follicles of cows, and to examine differentially expressed proteins in FF obtained in-vivo from preovulatory follicles of less fertile cows (also termed “repeat breeder”) and control (CTL) cows. The proteome of FF from 10 preovulatory follicles that were aspirated in vivo (estradiol/progesterone>1) was analyzed. This novel dataset contains 219 identified and quantified proteins in FF, consisting mainly of binding proteins, proteases, receptor ligands, enzymes and transporters. In addition, differential abundance of 8 proteins relevant to follicular function was found in LFC compared to CTL; these findings are discussed in our recent research article Zachut et al. (2016) [1]. The present dataset of bovine FF proteome can be used as a reference for any study involving disorders of follicular development in dairy cows or in comparative studies between species. PMID:27182550

  11. Insulin-like growth factor I in sera, ovarian follicles and follicular fluid of cows with spontaneous or induced cystic ovarian disease.

    PubMed

    Ortega, Hugo H; Palomar, Martin M; Acosta, Juan C; Salvetti, Natalia R; Dallard, Bibiana E; Lorente, Juan A; Barbeito, Claudio G; Gimeno, Eduardo J

    2008-06-01

    The objective of this research was to determine changes in IGF-I levels in serum and follicular fluid, and immunoreactivity of the follicle wall of cows with spontaneous (slaughter specimens) or ACTH-induced follicular cysts, and to compare results to normal cycling (control) cows after selection of the ovulatory follicle. Concentrations of IGF-I in serum did not differ between control and cystic animals (p=0.76). Fluid from the ovulatory follicle in control cows had 41% higher concentrations of IGF-I than that from cystic follicles collected at slaughter (spontaneous cysts; p<0.05) and 70% higher than that in induced follicular cysts (p<0.05). An intense positive immunostaining with anti-IGF-I was observed in granulosa cells (p<0.05) and in the theca interna (p<0.05) of secondary and tertiary follicles in all three groups of animals, but staining was less intense in cystic (p<0.05) and atretic follicles (p<0.05). This study provides evidence to suggest that cystic ovarian disease in cattle is associated with decreased concentrations of IGF-I in follicular fluid, but not in serum, and decreased production of IGF-I in the follicular wall. These data support the notion that IGF-I plays a role in the regulation of folliculogenesis, and may participate in the pathogenesis of cystic ovarian disease in cattle. PMID:17631370

  12. Comparison of bacterial endotoxin lipopolysaccharide concentrations in the blood, ovarian follicular fluid and uterine fluid: a clinical case of bovine metritis

    PubMed Central

    MAGATA, Fumie; ISHIDA, Yoshikazu; MIYAMOTO, Akio; FURUOKA, Hidefumi; INOKUMA, Hisashi; SHIMIZU, Takashi

    2014-01-01

    We investigated the concentration of the bacterial endotoxin lipopolysaccharide (LPS) in the blood, ovarian follicular fluid and uterine fluid of a clinical case of bovine metritis. A 2-year-old lactating Holstein cow exhibited continuous fever >39.5°C for more than 2 weeks after normal calving. The cow produced a fetid, watery, red-brown uterine discharge from the vagina and was diagnosed with metritis. The LPS concentrations in plasma and uterine fluid were 0.94 and 6.34 endotoxin units (EU)/ml, respectively. One of seven follicles showed an extremely high level of LPS (12.40 EU/ml) compared to the other follicles (0.62–0.97 EU/ml). These results might suggest the presence of high concentration of LPS in follicles in cows with postpartum metritis. PMID:25223344

  13. Comparison of bacterial endotoxin lipopolysaccharide concentrations in the blood, ovarian follicular fluid and uterine fluid: a clinical case of bovine metritis.

    PubMed

    Magata, Fumie; Ishida, Yoshikazu; Miyamoto, Akio; Furuoka, Hidefumi; Inokuma, Hisashi; Shimizu, Takashi

    2015-01-01

    We investigated the concentration of the bacterial endotoxin lipopolysaccharide (LPS) in the blood, ovarian follicular fluid and uterine fluid of a clinical case of bovine metritis. A 2-year-old lactating Holstein cow exhibited continuous fever >39.5°C for more than 2 weeks after normal calving. The cow produced a fetid, watery, red-brown uterine discharge from the vagina and was diagnosed with metritis. The LPS concentrations in plasma and uterine fluid were 0.94 and 6.34 endotoxin units (EU)/ml, respectively. One of seven follicles showed an extremely high level of LPS (12.40 EU/ml) compared to the other follicles (0.62-0.97 EU/ml). These results might suggest the presence of high concentration of LPS in follicles in cows with postpartum metritis. PMID:25223344

  14. Effect of ammonia-generating diet on ovine serum and follicular fluid ammonia and urea levels, serum oestrogen and progesterone concentrations and granulosa cell functions.

    PubMed

    Nandi, S; Mondal, S; Pal, D T; Gupta, P S P

    2016-04-01

    This study was undertaken to elucidate the effect of ammonia-generating diet on serum and follicular fluid ammonia and urea levels, serum oestrogen and progesterone concentrations and granulosa cell growth and secretion parameters in ewes (Ovis aries). Ewes were fed with 14% CP diet (control) or ammonia-generating diet or ammonia-generating diet plus soluble sugar. The serum and follicular fluid ammonia and urea level, serum oestrogen and progesterone levels and granulosa cell (obtained from ovaries of slaughtered ewes) growth parameters and secretory activities were estimated. Ammonia-generating diet (high-protein diet) increased the serum ammonia and urea concentration. Supplementation of soluble sugar significantly reduced the ammonia concentration in serum with comparable levels as in control group; however, the urea level in the same group was higher than that observed in control group. Supplementation of soluble sugar significantly reduced the follicular fluid ammonia concentration; however, the level was significantly higher compared to control group. Supplementation of soluble sugar brought down the follicular fluid urea level comparable to that observed in control group. Oestrogen and progesterone levels remained unchanged in ewes fed with different types of diet. Oestrogen and progesterone secretion were significantly lowered from granulosa cells recovered from ewes fed with high ammonia-generating diet. Low metabolic activity and high incidence of apoptosis were observed in granulosa cells obtained from ovaries of ewes fed with ammonia-generating diet. PMID:26211538

  15. Metabolic changes in follicular fluid of the dominant follicle in high-yielding dairy cows early post partum.

    PubMed

    Leroy, J L M R; Vanholder, T; Delanghe, J R; Opsomer, G; Van Soom, A; Bols, P E J; Dewulf, J; de Kruif, A

    2004-09-15

    Characteristics of the intrafollicular environment to which the preovulatory oocyte is exposed may be one of the major factors determining subsequent fertility. The aim of our study was to examine to what extent metabolic changes that occur in early post partum high-yielding dairy cows are reflected in the follicular fluid (FF) of the dominant follicle (>8 mm). Nine blood samples were taken per cow from nine high-yielding dairy cows between 7 days before and 46 days after parturition. From Day 14 post partum on and together with blood sampling, FF samples of the largest follicle were collected from the same cows by means of transvaginal follicle aspiration. Serum and FF samples were analyzed using commercial clinical and photometric chemistry assays for glucose, beta-hydroxybutyrate (beta-OHB), urea, total protein (TP), triglycerides (TG), non-esterified fatty acids (NEFA) and total cholesterol (TC). All cows lost body condition during the experimental period (0.94+/-0.09 points) illustrating a negative energy balance during the experimental period. In FF, glucose concentrations were significantly higher and the TP, TG, NEFA and TC concentrations were significantly lower than in serum (P<0.05). The concentrations of glucose, beta-OHB, urea and TC in serum and in FF changed significantly over time (P<0.05). Throughout the study, changes of all metabolites in serum were reflected by similar changes in FF. Especially for glucose, beta-OHB and urea the correlations were remarkably high. The results from the present study confirm that the typical metabolic adaptations which can be found in serum of high-yielding dairy cows shortly post partum, are reflected in follicular fluid and, therefore, may affect the quality of both the oocyte and the granulosa cells. PMID:15289052

  16. Development of a duplex quantitative polymerase chain reaction assay for detection of bovine herpesvirus 1 and bovine viral diarrhea virus in bovine follicular fluid.

    PubMed

    Marley, Mylissa S D; Givens, M Daniel; Galik, Patricia K; Riddell, Kay P; Stringfellow, David A

    2008-07-15

    The objective of this study was to develop a duplex quantitative polymerase chain reaction (qPCR) assay for simultaneous detection of bovine herpesvirus 1 (BoHV-1) and bovine viral diarrhea virus (BVDV) type I and type II. Follicular fluid was collected from a BoHV-1 acutely infected heifer, a BVDV I persistently infected heifer, and from 10 ovaries recovered from an abattoir. Both the BoHV-1 and BVDV contaminated follicular fluid were diluted 1:5 to 1:10(7) using the pooled, abattoir-origin follicular fluid. Each dilution sample was analyzed using the duplex qPCR, virus isolation, reverse transcription-nested PCR (RT-nPCR), and BoHV-1 qPCR. The duplex qPCR was able to simultaneously detect BoHV-1 and BVDV I in the fluid diluted to 1:100 and 1:1000, respectively. These results corresponded with the reverse transcription-nested PCR and BoHV-1 qPCR. Therefore, the duplex qPCR might be used for quality assurance testing to identify these two viruses in cells, fluids and tissues collected from donor animals and used in reproductive technologies. PMID:18452983

  17. The influence of the corpus luteum on metabolites composition of follicular fluid from different sized follicles and their relationship to serum concentrations in dairy cows.

    PubMed

    Shabankareh, Hamed Karami; Kor, Nasroallah Moradi; Hajarian, Hadi

    2013-08-01

    The presence of corpus luteum may have a local effect on metabolite composition of follicular fluid (FF) and could indirectly influence follicular development and oocyte quality. The purpose of this study was to examine the influence of the corpus luteum on metabolite composition of follicular fluid (FF), harvested from different-sized follicles and the relationship between metabolite composition of FF to blood serum in dairy cows. Ovaries and blood samples were collected from 30 female adult Holstein Friesian cows, 4-7 years old, with clinically normal reproductive tracts. The animals were in the diestrus stage and selected post mortem. The ovaries collected were classified based on the presence and absence of corpus luteum (CL(+/-)). Visible follicles on the surface of the ovaries were classified into (i) small (3-5mm), (ii) medium (6-9 mm) and (iii) large (10-20mm) based on their diameter. Follicular fluid was aspirated from follicles with different sizes in CL(+) and CL(-) ovaries. Blood and FF samples were analyzed for various biochemical constituents including glucose, cholesterol, triglyceride, total protein, albumin and globulin. The results showed that serum concentration of glucose, cholesterol and triglyceride was significantly different (p≤0.05) in FF from follicles of different size categories. Differences between various follicle size categories in CL(-) ovaries were only significant for concentrations of glucose, cholesterol and triglyceride. FF concentration of glucose and cholesterol in the same follicle size categories in CL(+) ovaries was significantly lower than that of CL(-) ovaries. These results indicate that levels of the biochemical metabolites in serum and FF differ significantly. In addition, FF concentrations of biochemical metabolites were related to follicular size and to the presence or absence of corpus luteum. PMID:23890803

  18. Human GV oocytes generated by mitotically active germ cells obtained from follicular aspirates

    PubMed Central

    Ding, Xinbao; Liu, Guishu; Xu, Bo; Wu, Changqing; Hui, Ning; Ni, Xin; Wang, Jian; Du, Meirong; Teng, Xiaoming; Wu, Ji

    2016-01-01

    Human female germline stem cells (FGSCs) have opened new opportunities for understanding human oogenesis, delaying menopause, treating infertility, and providing a new strategy for preserving fertility. However, the shortage of adult human ovaries tissues available impedes their future investigations and clinical applications. Here, we have established FGSC lines from scarce ovarian cortical tissues that exist in follicular aspirates (faFGSCs), which are produced and discarded in in vitro fertilization centers worldwide. The faFGSCs have characteristics of germline stem cells involved in the gene expression profile, growth characteristics, and a normal karyotype consistent with that of FGSCs obtained from ovarian cortexes surgically removed from patients (srFGSCs). Furthermore, faFGSCs have developmental potentials including spontaneous differentiation into oocytes under feeder-free conditions, communicating with granulosa cells by gap junctions and paracrine factors, entering meiosis after RA induction, as well as forming follicles after injection into human ovarian cortical tissues xenografted into adult immunodeficient female mice. Lastly, we developed a strategy guiding FGSCs differentiated into germinal vesicle (GV) stage oocytes in vitro and revealed their developmental mechanisms. Our study not only provides a new approach to obtain human FGSCs for medical treatment, but also opens several avenues to investigate human oogenesis in vitro. PMID:27357640

  19. Human GV oocytes generated by mitotically active germ cells obtained from follicular aspirates.

    PubMed

    Ding, Xinbao; Liu, Guishu; Xu, Bo; Wu, Changqing; Hui, Ning; Ni, Xin; Wang, Jian; Du, Meirong; Teng, Xiaoming; Wu, Ji

    2016-01-01

    Human female germline stem cells (FGSCs) have opened new opportunities for understanding human oogenesis, delaying menopause, treating infertility, and providing a new strategy for preserving fertility. However, the shortage of adult human ovaries tissues available impedes their future investigations and clinical applications. Here, we have established FGSC lines from scarce ovarian cortical tissues that exist in follicular aspirates (faFGSCs), which are produced and discarded in in vitro fertilization centers worldwide. The faFGSCs have characteristics of germline stem cells involved in the gene expression profile, growth characteristics, and a normal karyotype consistent with that of FGSCs obtained from ovarian cortexes surgically removed from patients (srFGSCs). Furthermore, faFGSCs have developmental potentials including spontaneous differentiation into oocytes under feeder-free conditions, communicating with granulosa cells by gap junctions and paracrine factors, entering meiosis after RA induction, as well as forming follicles after injection into human ovarian cortical tissues xenografted into adult immunodeficient female mice. Lastly, we developed a strategy guiding FGSCs differentiated into germinal vesicle (GV) stage oocytes in vitro and revealed their developmental mechanisms. Our study not only provides a new approach to obtain human FGSCs for medical treatment, but also opens several avenues to investigate human oogenesis in vitro. PMID:27357640

  20. The correlation between follicular fluid pregnancy-associated plasma protein A levels, fertilization, and embryo quality in GnRH agonist and GnRH antagonist protocols in ART cycles

    PubMed Central

    Dehghani Firouzabadi, Razieh; Mohammadian, Farnaz; Mashayekhy, Mehri; Davar, Robab; Eftekhar, Maryam

    2012-01-01

    Background: Determination of oocyte fertilization and embryo quality are one of the most important purposes in ART cycles. Follicular fluid provides an important microenvironment for development of oocytes and some biochemical characteristics of the follicular fluid, such as pregnancy-associated plasma protein-A (PAPP-A), may play an important role in prediction of success rate of ART. Objective: This study was performed to evaluate whether there was any difference in follicular fluid PAPP-A, fertilization, and embryo quality between GnRH agonist long protocol and flexible GnRH antagonist multiple-dose protocol in ART cycles. Materials and Methods: A total of 100 women who were candidates for ART were enrolled the study and were divided into two groups, GnRH agonist (GnRHa) long protocol (n=51) and flexible GnRH antagonist (GnRHant) multiple-dose protocol (n=49). Follicular fluid sample was obtained from a single mature follicle and follicular fluid PAPP-A level, fertilization and embryo quality of the same oocyte were evaluated in both groups. Results: There was no significant difference in the mean levels of follicular fluid PAPP-A between the GnRHa protocol and GnRHant protocol (3.5±1.4 vs. 3.8±1.9, respectively). The mean levels of follicular fluid PAPP-A in fertilized oocyte and good quality embryo were comparable in GnRHa and GnRHant protocols. Conclusion: Our data indicated that no differences of follicular fluid PAPP-A levels were observed between cycles using GnRHa long protocol and those of using flexible GnRHant multiple-dose protocol. PMID:25246915

  1. Bioanalytical validation for simultaneous quantification of non-aromatic steroids in follicular fluid from cattle via ESI-LC-MS/MS.

    PubMed

    Kunze, Martin; Wirthgen, Elisa; Walz, Christina; Spitschak, Marion; Brenmoehl, Julia; Vanselow, Jens; Schwerin, Manfred; Wimmers, Klaus; Hoeflich, Andreas

    2015-12-15

    The family of steroid hormones is quite attractive for the approach of phenotype monitoring in farm animals. Therefore, we developed a new protocol for the quantitative analysis of natural steroids in follicular fluid from dairy cows. The corresponding steroid profile, which consists of progesterone, corticosterone, hydrocortisone, testosterone, and androstenedione covering three distinct steroid classes, was determined by LC/MS. Quantification is achieved by use of steroid standards diluted in steroid-free follicular fluid as calibrators. Thus, the new protocol does not require deuterated standards. In order to correct for conditional performance of the analytical system we have used dexamethasone as an internal standard. The method was validated according to EMA guidelines. Within- and between-day variations were below 20% for most parameters assessed. All steroids assessed had lower limits of quantification in the range of 2.1 to 4.4ng/ml. We have established a simple and sensitive analytical system in order to step towards a broader and cost-efficient phenotyping analysis in follicular fluid from dairy cows. PMID:26600283

  2. An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm.

    PubMed

    Leemans, Bart; Gadella, Bart M; Stout, Tom A E; Nelis, Hilde; Hoogewijs, Maarten; Van Soom, Ann

    2015-09-01

    Induction of hyperactivated motility is considered essential for triggering the release of oviduct-bound mammalian spermatozoa in preparation for fertilization. In this study, oviduct-bound stallion spermatozoa were exposed for 2 h to: i) pre-ovulatory and ii) post-ovulatory oviductal fluid; iii) 100% and iv) 10% follicular fluid (FF); v) cumulus cells, vi) mature equine oocytes, vii) capacitating and viii) non-capacitating medium. None of these triggered sperm release or hyperactivated motility. Interestingly, native FF was detrimental to sperm viability, an effect that was negated by heat inactivation, charcoal treatment and 30 kDa filtration alone or in combination. Moreover, sperm suspensions exposed to treated FF at pH 7.9 but not pH 7.4 showed Ca(2+)-dependent hypermotility. Fluo-4 AM staining of sperm showed elevated cytoplasmic Ca(2+) in hyperactivated stallion spermatozoa exposed to treated FF at pH 7.9 compared to a modest response in defined capacitating conditions at pH 7.9 and no response in treated FF at pH 7.4. Moreover, 1 h incubation in alkaline, treated FF induced protein tyrosine phosphorylation in 20% of spermatozoa. None of the conditions tested induced widespread release of sperm pre-bound to oviduct epithelium. However, the hyperactivating conditions did induce release of 70-120 spermatozoa per oviduct explant, of which 48% showed protein tyrosine phosphorylation and all were acrosome-intact, but capable of acrosomal exocytosis in response to calcium ionophore. We conclude that, in the presence of elevated pH and extracellular Ca(2+), a heat-resistant, hydrophilic, <30 kDa component of FF can trigger protein tyrosine phosphorylation, elevated cytoplasmic Ca(2+) and hyperactivated motility in stallion sperm, but infrequent release of sperm pre-bound to oviduct epithelium. PMID:26242588

  3. Expression of receptors for VEGFs on normal human thyroid follicular cells and their role in follicle formation.

    PubMed

    Susarla, Radhika; Gonzalez, Ana-Maria; Watkinson, John C; Eggo, Margaret C

    2012-05-01

    Human thyroid follicular cells in culture expressed the mRNAs for the receptors for vascular endothelial growth factors (VEGFRs). The relative expression was neuropilin1 = neuropilin2 = VEGFR2 > VEGFR1 > VEGFR3. Western blotting for VEGFR2 showed labeling of proteins ~200-230 kDa. Clonal follicular thyroid cell lines (FRTL5 and FTC133) also expressed mRNAs for the VEGFR1 and 2 obviating concerns of endothelial cell contamination. In the primary cultures, TSH, which is essential for expression of differentiated function, reduced VEGFR2 mRNA levels by 60%. Immunostaining for VEGFRs and neuropilin2 (NRP2), showed expression on the plasma membrane but with the exception of neuropilin1 (NRP1), all VEGFRs were also found in the cytoplasm and nucleus. Antibody specific for phosphotyrosine 1214 in VEGFR2 showed that the receptor was phosphorylated in the primary cultures and the cell lines. When VEGFR signaling was blocked with a specific inhibitor, follicle formation in the primary cultures was enhanced suggesting that VEGFR activation was detrimental to follicle formation. Immunostaining of sections of normal thyroids and various pathologies showed staining for VEGFR2 and pVEGFR2. We conclude that normal thyroid follicular cell express VEGFRs. For VEGFR2 its subcellular localization suggests functions additional to that of a cell surface receptor and a role in follicular integrity. PMID:21751212

  4. Characterization of human follicular thyroid cancer cell lines in preclinical mouse models

    PubMed Central

    Reeb, Ashley N; Ziegler, Andrea

    2016-01-01

    Follicular thyroid cancer (FTC) is the second most common type of thyroid cancers. In order to develop more effective personalized therapies, it is necessary to thoroughly evaluate patient-derived cell lines in in vivo preclinical models before using them to test new, targeted therapies. This study evaluates the tumorigenic and metastatic potential of a panel of three human FTC cell lines (WRO, FTC-238, and TT1609-CO2) with defined genetic mutations in two in vivo murine models: an orthotopic thyroid cancer model to study tumor progression and a tail vein injection model to study metastasis. All cell lines developed tumors in the orthotopic model, with take rates of 100%. Notably, WRO-derived tumors grew two to four times faster than tumors arising from the FTC-238 and TT2609-CO2 cell lines. These results mirrored those of a tail vein injection model for lung metastasis: one hundred percent of mice injected with WRO cells in the tail vein exhibited aggressive growth of bilateral lung metastases within 35 days. In contrast, tail vein injection of FTC-238 or TT2609-CO2 cells did not result in lung metastasis. Together, our work demonstrates that these human FTC cell lines display highly varied tumorigenic and metastatic potential in vivo with WRO being the most aggressive cell line in both orthotopic and lung metastasis models. This information will be valuable when selecting cell lines for preclinical drug testing. PMID:26830329

  5. Follicular Helper CD4+ T Cells in Human Neuroautoimmune Diseases and Their Animal Models

    PubMed Central

    Fan, Xueli; Lin, Chenhong; Han, Jinming; Jiang, Xinmei; Zhu, Jie; Jin, Tao

    2015-01-01

    Follicular helper CD4+ T (TFH) cells play a fundamental role in humoral immunity deriving from their ability to provide help for germinal center (GC) formation, B cell differentiation into plasma cells and memory cells, and antibody production in secondary lymphoid tissues. TFH cells can be identified by a combination of markers, including the chemokine receptor CXCR5, costimulatory molecules ICOS and PD-1, transcription repressor Bcl-6, and cytokine IL-21. It is difficult and impossible to get access to secondary lymphoid tissues in humans, so studies are usually performed with human peripheral blood samples as circulating counterparts of tissue TFH cells. A balance of TFH cell generation and function is critical for protective antibody response, whereas overactivation of TFH cells or overexpression of TFH-associated molecules may result in autoimmune diseases. Emerging data have shown that TFH cells and TFH-associated molecules may be involved in the pathogenesis of neuroautoimmune diseases including multiple sclerosis (MS), neuromyelitis optica (NMO)/neuromyelitis optica spectrum disorders (NMOSD), and myasthenia gravis (MG). This review summarizes the features of TFH cells, including their development, function, and roles as well as TFH-associated molecules in neuroautoimmune diseases and their animal models. PMID:26300592

  6. Dysregulation of autophagy in human follicular lymphoma is independent of overexpression of BCL-2

    PubMed Central

    McCarthy, Aine; Marzec, Jacek; Clear, Andrew; Petty, Robert D.; Coutinho, Rita; Matthews, Janet; Wilson, Andrew; Iqbal, Sameena; Calaminici, Maria; Gribben, John G.; Jia, Li

    2014-01-01

    Overexpression of the anti-apoptotic protein BCL-2 is characteristic of human follicular lymphoma (FL) and some cases of diffuse large B cell lymphoma (DLBCL). We aimed to determine autophagy status in primary FL and DLBCL samples and the BCL-2+/BCL-2− lymphoma cell lines using both autophagy PCR array and tissue microarray (TMA). A greater number of autophagy machinery genes were up-regulated in the BCL-2+ Su-DHL4 cell line compared with BCL-2− Su-DHL8 cells, at both the basal level and in response to autophagic stress. The autophagy-related gene expression profiles were determined in purified and unpurified malignant human lymph node biopsies. Seven autophagy machinery genes were up-regulated in purified FL B-cells compared with reactive B-cells. Only 2 autophagy machinery genes were up-regulated in DLBCL B-cells. In unpurified tissue biopsies, 20 of 46 genes in FL and 2 of 5 genes in DLBCL with increased expression were autophagy machinery genes. Expression of autophagy substrates p62 and LC3 were determined by TMAs. FL samples showed significantly decreased levels of both p62 and LC3 compared with reactive and DLBCL, indicative of an increased autophagy activity in FL. In summary, these results demonstrate that FL showed increased basal autophagy activity, regardless of overexpression of BCL-2 in this disease. PMID:25362242

  7. Exploratory study of the association of steroid profiles in stimulated ovarian follicular fluid with outcomes of IVF treatment.

    PubMed

    Kushnir, Mark M; Naessén, Tord; Wanggren, Kjell; Hreinsson, Julius; Rockwood, Alan L; Meikle, A Wayne; Bergquist, Jonas

    2016-09-01

    Steroid concentrations in stimulated follicular fluid (sFF) samples have been linked to the quality of oocytes used in IVF treatments. Most of the published studies focused on evaluating the association of the IVF outcomes with only a few of the steroids, measured by immunoassays (IA). We performed a treatment outcome, prospective cohort study using stimulated FF sampled from 14 infertile women undergoing IVF treatment; single oocyte was used per IVF cycle. Fourteen endogenous steroids were analyzed in 22 ovarian follicle aspirations, which corresponded to the embryos used in the IVF. Ten oocytes were associated with live birth (LB) and 12 with no pregnancy (NP). Steroids were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. Differences in distribution of concentrations in association with the pregnancy outcome (LB or NP), and receiver operating characteristic (ROC) curves analysis were performed for the entire cohort and for within-women data. The predominant androgen and estrogen in stimulated sFF were androstenedione (A4) and estradiol (E2), respectively. Lower concentrations of pregnenolone (Pr), lower ratios of A4/ dehydroepiandrosterone (DHEA), testosterone (Te)/DHEA, and greater ratios of E2/Te, and estrone/A4 were observed in sFF samples associated with LB. Among the oocytes associated with NP, in four out of 12 samples total concentration of androgens was above the distribution of the concentrations in the oocytes corresponding to the LB group. Observations of the study indicated increased consumption of precursors and increased biosynthesis of estrogens in the follicles associated with LB. Our data suggest that potentially steroid profiles in sFF obtained during oocyte retrieval may serve as biomarkers for selection of the best embryo to transfer after IVF. PMID:26388251

  8. Anti-laminin-1 antibodies in serum and follicular fluid of women with Hashimoto's thyroiditis undergoing in vitro fertilization.

    PubMed

    Caccavo, Domenico; Pellegrino, Nelly M; Nardelli, Claudia; Vergine, Silvia; Leone, Luca; Marolla, Alessandra; Vacca, Margherita P; Depalo, Raffaella

    2016-06-01

    The aim of this study is to evaluate the presence of anti-laminin-1 antibodies (aLN-1) in sera and follicular fluid (FF) of infertile women affected by Hashimoto's thyroiditis (HT) undergoing in vitro fertilization (IVF) and its impact on oocyte maturation and IVF outcome. aLN-1 were measured by a home-made enzyme linked immunosorbent assay (ELISA) in: (1) sera and FF from 44 infertile women affected by HT (HTIW) with tubal factor or male factor as primary cause of infertility; (2) in sera and FF from 28 infertile women without HT, with tubal factor or male factor as cause of infertility (infertile controls-ICTR); and (3) in sera from 50 fertile women (FW). aLN-1 serum levels were significantly higher in HTIW when compared with both fertile women and ICTR (P <0.001and P <0.01, respectively). Assuming as cutoff the 99th percentile of values obtained in sera of FW, 43.2% of HTIW and 3.6% of ICTR were aLN-1 positive (P = 0.0001). Also aLN-1 detected in FF from HTIW were significantly higher in comparison with those found in FF of ICTR (P = 0.006). In HTIW, metaphase II oocyte count showed inverse correlation with both serum and FF aLN-1 levels (r = 0.34, P = 0.02 and r = 0.33, P = 0.03, respectively). Implantation and pregnancy rates were significantly lower in HTIW (7.9% and 9.1%, respectively) when compared with ICTR (23% and 31.1%, respectively) (P = 0.015 and P = 0.03, respectively). Our results demonstrated for the first time the presence of aLN-1 in a relevant percentage of HTIW and suggest that these auto-antibodies may impair IVF outcome. PMID:26813862

  9. DNA rearrangement in human follicular lymphoma can involve the 5' or the 3' region of the bcl-2 gene

    SciTech Connect

    Tsujimoto, Y.; Bashir, M.M.; Givol, I.; Cossman, J.; Jaffe, E.; Croce, C.M.

    1987-03-01

    In most human lymphomas, the chromosome translocation t(14;18) occurs within two breakpoint clustering regions on chromosome 18, the major one at the 3' untranslated region of the bcl-2 gene and the minor one at 3' of the gene. Analysis of a panel of follicular lymphoma DNAs using probes for the first exon of the bcl-2 gene indicates that DNA rearrangements may also occur 5' to the involved bcl-2 gene. In this case the IgH locus and the bcl-2 gene are found in an order suggesting that an inversion also occurred during the translocation process. The coding region of the bcl-2 gene, however, are left intact in all cases of follicular lymphoma studied to date.

  10. The assesment of follicular fluid presepsin levels in poor ovarian responder womenandits relationship with the reproductive outcomes

    PubMed Central

    Ovayolu, Ali; Özdamar, Özkan; Gün, İsmet; Arslanbuğa, Cansev Y; Kutlu, Tayfun; Tunalı, Gülden; Uluhan, Ramazan

    2015-01-01

    A considerable proportion of all women undergoing IVFrespond poorly to gonadotropin stimulation. These women are reported to be associated with increased cancellation rates and lower pregnancy rates. It has been hypothesized that poor response to ovarian stimulation is a first sign of ovarian ageing or premature ovarian failure, which might be related to altered inflammatory response in the body. We aimed to compare follicular fluid presepsin levels between poor- and normo-responder patients to ovarian stimulation, to assess its relationship with reproductive outcomes. This study included infertility patients who underwent ovulation induction with either long GnRH agonist or GnRH antagonist protocols and who subsequently underwent IVF/ICSI. Included patients were assigned to two groups according to the Bologna criteria for poor ovarian response. Group 1 and 2 consisted of normo- and poor-responder patients, respectively.The 2 groups were compared in terms of FF presepsin levels. Also, any relationship between the FF presepsin levels and fertility outcomes was assessed within the groups. The groups were compared by using student’s t-test, Mann-Whitney U test and X2 test, where appropriate. Pregnancy rates were not significantly different between the groups (22.6% and 17.6%; P=0.650, respectively). FF presepsin levels were higher in Group 1, however, the difference was not statistically significant (298.0±797.4 and 149.2±422.3; P=0.190, respectively). FF presepsin levels did not significantly differ between pregnancy positive and the pregnancy negative patients in both Group 1 (243.6±531.1 and 314.3±866.5; P=0.055, respectively) and Group 2 (112.2±79.8 and 157.1±464.3; P=0.394, respectively). Consequently, FF presepsin seems not to be a reliable marker in predicting pregnancy in both normo-responder and poor-responder infertility groups. PMID:26309683

  11. Paraoxonase (PON) 1, 2 and 3 expression in granulosa cells and PON1 activity in follicular fluid of dairy cows.

    PubMed

    Schneider, A; Absalon-Medina, V A; Esposito, G; Corrêa, M N; Butler, W R

    2013-12-01

    Normal metabolic activity in ovarian follicles may result in oxidative stress and damage to oocytes. The aim of this study was to evaluate expression of the natural anti-oxidants paraoxonase (PON) 1, 2 and 3 in granulosa cells and PON1 activity in follicular fluid (FF) and plasma of dairy cows. For the first experiment, ovaries were collected from cows at slaughter, after which follicles were dissected and classified as oestrogen active (EAF) or atretic (ATF). Expression of PON1, PON2 and PON3 mRNA was evaluated in granulosa cells, and activity of PON1 was measured in FF. PON1 mRNA was undetectable in granulosa cells, PON2 mRNA expression was not different between follicle types, and PON3 mRNA tended to be higher in EAF (p = 0.11). The activity of PON1 in FF was higher (p = 0.01) for EAF (82.6 ± 8.0 kU/L) than ATF (53.9 ± 6.8 kU/L), as were high-density lipoproteins (HDL), low-density lipoproteins (LDL) and total cholesterol concentrations. In the second experiment, we aimed to compare plasma and FF PON1 activity in early lactation Holstein cows (n = 15) with pre-ovulatory EAF. Activity of PON1 was twofold higher (p < 0.0001) in plasma (122.5 ± 11.1 kU/L) than in FF (61.4 ± 5.2 kU/L). Plasma concentrations were also higher (p < 0.0001) for HDL, LDL and total cholesterol when compared to FF. In conclusion, FF concentrations of PON1, HDL, LDL and total cholesterol were higher in healthy oestrogen active bovine follicles than in atretic follicles. PON1 was not expressed by granulosa cells indicating that high PON1 activity in bovine FF is apparently derived by transfer from blood in association with HDL. PMID:23758560

  12. [Erythropoietin levels in human tear fluid].

    PubMed

    Zakharov, Iu M; Rykun, V S; Bagautdinov, D E; Romanenko, E A

    2010-11-01

    Erythropoietin level was evaluated in blood plasma and tear fluid of humans with normal functions of eye and normal blood characteristics. We examined 21 patients. Principle ability of erythropoietin level detection in patient's tear fluid ascertained. PMID:21427970

  13. Origin, Possible Function and Fate of “Follicular Cells” in the Anterior Lobe of the Human Pituitary

    PubMed Central

    Horvath, E.; Kovacs, K.; Penz, G.; Ezrin, C.

    1974-01-01

    In human anterior pituitaries, follicular structures were found to develop by transformation of various types of glandular cells around foci of ruptured granulated cells undergoing destruction. In phase I, junctional complexes between granulated cells, as well as microvilli at the luminal surfaces of cell membranes, are formed. In phase II, degranulation and dedifferentiation of cytoplasm dominate the picture. Phase III follicular cells are practically devoid of secretory granules and other ultrastructural features characteristic of granulated cells. The participation of cell types in follicle formation does not appear to be limited. The follicular content in phase I-II is clearly recognizable as cellular debris from adenohypophysiocytes. These findings indicate that neither follicles nor junctional complexes are necessarily permanent structures of the anterior pituitary. It can be assumed that substances escaping from ruptured granulated cells may induce the formation of junctional complexes between adjacent cells. ImagesFig 5Fig 6Fig 7Fig 8Fig 9Fig 1Fig 2Fig 10Fig 3Fig 4 PMID:4447128

  14. Systemic Inflammation Is Associated with Ovarian Follicular Dynamics during the Human Menstrual Cycle

    PubMed Central

    Clancy, Kathryn B. H.; Baerwald, Angela R.; Pierson, Roger A.

    2013-01-01

    Background Ovarian processes and the timing of ovulation are important predictors of both female fertility and reproductive pathology. Multiple waves of antral follicular development have been documented during the menstrual cycle in women. However, the mechanisms underlying the development of follicular waves and their clinical significance are not fully understood. The objective of this study was to examine the relationship between C-reactive protein (CRP) and follicular waves in healthy women. We wanted to determine whether follicular wave dynamics influence systemic inflammation, as ovarian activity increases local inflammatory processes and blood flow. We tested the hypothesis that women with 3 follicular waves would have higher CRP concentrations than those with 2 waves. We further hypothesized that a greater number of major waves (those with a dominant follicle) would be positively associated with CRP. Methods/Principal Findings Thirty-nine healthy women underwent daily transvaginal ultrasound examinations for one interovulatory interval, as part of an earlier study. Serum was collected every 3 days during the interovulatory interval (IOI). Enzyme-linked immunosorbent assays were conducted to quantify serum CRP concentrations. Women with 3 waves had higher average log CRP concentrations (n = 14, −0.43±0.35) over the IOI than those with 2 waves (n = 25, −0.82±0.47, p = 02). Average log CRP concentrations were greater in women with 3 (0.30±0.31) versus 1 (−0.71±0.55) or 2 (−0.91±0.47) major waves (p = 0.03). Greater average CRP over the IOI was attributed to greater CRP in the follicular, but not the luteal phase, of the IOI. Conclusions/Significance A greater number of total antral follicular waves, in particular major waves, corresponded to greater serum concentrations of CRP. These findings suggest that women with a greater number of follicular waves exhibit greater tissue remodeling and therefore greater local and systemic

  15. The 'follicular trochanter': an epithelial compartment of the human hair follicle bulge region in need of further characterization.

    PubMed

    Tiede, S; Kloepper, J E; Whiting, D A; Paus, R

    2007-11-01

    Recent articles on hair follicle stem cells have summarized the current state of knowledge of what has been termed the hair follicle 'bulge'. During the course of immunohistological studies aimed at characterizing the expression of selected extracellular matrix proteins in the - as yet insufficiently characterized - niche of human bulge hair follicle stem cells, we have recently come across a largely forgotten, peculiar epithelial protrusion of the outer root sheath, which was visible in only a minority of all examined hair follicles. The morphology and immunoreactivity patterns of this structure, the 'follicular trochanter', are described herein. PMID:17714535

  16. Follicular dermographism.

    PubMed

    Shelley, W B; Shelley, E D

    1983-09-01

    Four patients who complained of an inexplicable pruritus of several weeks to several years in duration are presented. General medical studies were unremarkable and the skin was normal except for erythematous papular lesions at sites recently scratched. The skin changes could be reproduced by multiple firm strokes with the broad edge of a tongue blade. The transitory, discrete, follicular, urticarial papules so elicited have been labeled by us as follicular dermographism. It is an entity to be distinguished from cholinergic and aquagenic urticaria. We postulate that follicular dermographism occurs during periods of transient antigenemia. At such times, the frictional force of scratching or the tongue blade test releases antigen from the blood stream to trigger focal urticaria at sites of high density mast cells, namely around the hair follicle. Hydroxyzine is an effective therapy. PMID:6627988

  17. Matrix metalloproteinases 2 and 9 and their tissue inhibitors in the follicular fluid of patients with polycystic ovaries undergoing in vitro fertilisation.

    PubMed

    Baka, Stavroula; Zourla, Konstantina; Kouskouni, Evangelia; Makrakis, Evangelos; Demeridou, Stella; Tzanakaki, Despoina; Hassiakos, Dimitris; Creatsas, George

    2010-01-01

    The present study was undertaken to investigate the levels of matrix metalloproteinase (MMP)-2, MMP-9 and their tissue inhibitors (TIMP-2 and TIMP-1, respectively) in the follicular fluid of 39 patients with polycystic ovary syndrome (PCOS) and compare them with the levels found in 56 age- and weight-matched normally ovulating women, all undergoing in vitro fertilisation (IVF) treatment. Significantly higher levels of MMP-2 and MMP-9 (p=0.02 and p<0.001, respectively) as well as TIMP-2 and TIMP-1 (p=0.006 and p<0.001, respectively) were found in the PCOS group compared to controls. Women who achieved pregnancy had higher TIMP-1 levels compared to the non-pregnant ones in the control group (p=0.01). In conclusion, women with PCOS exhibited significantly increased gelatinolytic activity compared with controls of similar age and body mass index, thus indicating a more intense extracellular matrix remodelling in this group of patients during IVF treatment due to multiple follicular development and cyst formation. PMID:20555001

  18. Bilateral follicular cysts in a water buffalo.

    PubMed

    Khan, F A; Nabi, S U; Pande, Megha; Das, G K; Sarkar, M

    2011-03-01

    The present short communication puts on record a case of bilateral, multiple follicular cysts in a water buffalo along with a detailed description of its ovarian biometry and follicular fluid composition. The ovarian weight and biometrical parameters were much higher than in normal cycling buffaloes. A total of three follicular cysts were observed, two on the right ovary and one on the left ovary, measuring 4.9, 3.0 and 2.6 cm yielding 21, 9 and 5 ml of follicular fluid, respectively. The cystic fluid was deep yellow in colour with a viscous consistency. The follicular fluid concentrations of glucose, total protein, cholesterol, acid phosphatase, calcium, phosphorus and progesterone in all the cysts were within the range reported previously in normal buffalo follicular fluid; however, the alkaline phosphatase concentration in cyst 1 and total bilirubin concentration in cysts 1 and 2 were higher than the values in normal follicular fluid. In contrast, the levels of urea nitrogen in cysts 1 and 3, and oestradiol in cyst 3 were lower than the normal values. All the three follicles had an oestradiol to progesterone ratio less than 1. The results of our study suggest that follicular cysts in buffalo are oestrogenically inactive and have an altered concentration of certain biochemical and hormonal constituents. PMID:21086160

  19. Transcriptional Profiling of Human Endocervical Tissues Reveals Distinct Gene Expression in the Follicular and Luteal Phases of the Menstrual Cycle.

    PubMed

    Yildiz-Arslan, Sevim; Coon, John S; Hope, Thomas J; Kim, J Julie

    2016-06-01

    The endocervix plays an important role in providing appropriate protective mechanisms of the upper female reproductive tract (FRT) while at the same time providing the appropriate milieu for sperm transport. Hormone fluctuations throughout the menstrual cycle contribute to changes in the mucosal environment that render the FRT vulnerable to infectious diseases. The objective of this study was to identify genes in human endocervix tissues that were differentially expressed in the follicular versus the luteal phases of the menstrual cycle using gene expression profiling. A microarray using the IIlumina platform was performed with eight endocervix tissues from follicular and four tissues from luteal phases of the menstrual cycle. Data analysis revealed significant differential expression of 110 genes between the two phases, with a P value <0.05 and a fold change cutoff of 1.5. Categorization of these genes, using Ingenuity Pathway Analysis, MetaCore from Thomson Reuters, and DAVID, revealed genes associated with extracellular matrix remodeling and cell-matrix interactions, amino acid metabolism, and lipid metabolism, as well as immune regulation in the follicular phase tissues. In luteal phase tissues, genes associated with chromatin remodeling, inflammation, angiogenesis, oxidative stress, and immune cell regulation were predominately expressed. Using samples from additional patients' tissues, select genes were confirmed by quantitative real-time PCR; immunohistochemical staining was also done to examine protein levels. This is the first microarray analysis comparing gene expression in endocervix tissues in cycling women. This study identified key genes and molecular pathways that were differentially regulated during the menstrual cycle. PMID:27170437

  20. CD10 delineates a subset of human IL-4 producing follicular helper T cells involved in the survival of follicular lymphoma B cells.

    PubMed

    Amé-Thomas, Patricia; Hoeller, Sylvia; Artchounin, Catherine; Misiak, Jan; Braza, Mounia Sabrina; Jean, Rachel; Le Priol, Jérôme; Monvoisin, Céline; Martin, Nadine; Gaulard, Philippe; Tarte, Karin

    2015-04-01

    In follicular lymphoma (FL), follicular helper T cells (TFH) have been depicted as one of the main components of the malignant B-cell niche and a promising therapeutic target. Although defined by their capacity to sustain FL B-cell growth together with specific gene expression and cytokine secretion profiles, FL-TFH constitute a heterogeneous cell population. However, specific markers reflecting such functional heterogeneity are still lacking. In this study, we demonstrate that CD10 identifies a subset of fully functional germinal center TFH in normal secondary lymphoid organs. Importantly, this subset is amplified in the FL context, unlike in other B-cell lymphomas with a follicular growth pattern. Furthermore, whereas FL-TFH produce high levels of interleukin (IL)-21 and low levels of IL-17 irrespectively of their CD10 expression, CD10(pos) FL-TFH specifically exhibit an IL-4(hi)IFN-γ(lo)TNF-α(hi) cytokine profile associated with a high capacity to sustain directly and indirectly malignant B-cell survival. Altogether, our results highlight the important role of this novel functional subset in the FL cell niche. PMID:25733581

  1. Circulating follicular T helper cells and cytokine profile in humans following vaccination with the rVSV-ZEBOV Ebola vaccine.

    PubMed

    Farooq, Fouzia; Beck, Kevin; Paolino, Kristopher M; Phillips, Revell; Waters, Norman C; Regules, Jason A; Bergmann-Leitner, Elke S

    2016-01-01

    The most recent Zaire Ebolavirus (ZEBOV) outbreak was the largest and most widespread in recorded history, emphasizing the need for an effective vaccine. Here, we analyzed human cellular immune responses induced by a single dose of the rVSV-ZEBOV vaccine candidate, which showed significant protective efficacy in endemic populations in Guinea. This is the first in-depth characterization of ZEBOV-GP specific, circulating follicular T cells (cTfh). Since antibody titers correlated with protection in preclinical models of ZEBOV infection, Tfh were predicted to correlate with protection. Indeed, the ZEBOV-specific cTfh data correlated with antibody titers in human vaccines and unexpectedly with the Tfh17 subset. The combination of two cutting edge technologies allowed the immuno-profiling of rare cell populations and may help elucidate correlates of protection for a variety of vaccines. PMID:27323685

  2. Circulating follicular T helper cells and cytokine profile in humans following vaccination with the rVSV-ZEBOV Ebola vaccine

    PubMed Central

    Farooq, Fouzia; Beck, Kevin; Paolino, Kristopher M.; Phillips, Revell; Waters, Norman C.; Regules, Jason A.; Bergmann-Leitner, Elke S.

    2016-01-01

    The most recent Zaire Ebolavirus (ZEBOV) outbreak was the largest and most widespread in recorded history, emphasizing the need for an effective vaccine. Here, we analyzed human cellular immune responses induced by a single dose of the rVSV-ZEBOV vaccine candidate, which showed significant protective efficacy in endemic populations in Guinea. This is the first in-depth characterization of ZEBOV-GP specific, circulating follicular T cells (cTfh). Since antibody titers correlated with protection in preclinical models of ZEBOV infection, Tfh were predicted to correlate with protection. Indeed, the ZEBOV-specific cTfh data correlated with antibody titers in human vaccines and unexpectedly with the Tfh17 subset. The combination of two cutting edge technologies allowed the immuno-profiling of rare cell populations and may help elucidate correlates of protection for a variety of vaccines. PMID:27323685

  3. Human tonsil-derived follicular dendritic-like cells are refractory to human prion infection in vitro and traffic disease-associated prion protein to lysosomes.

    PubMed

    Krejciova, Zuzana; De Sousa, Paul; Manson, Jean; Ironside, James W; Head, Mark W

    2014-01-01

    The molecular mechanisms involved in human cellular susceptibility to prion infection remain poorly defined. This is due, in part, to the absence of any well characterized and relevant cultured human cells susceptible to infection with human prions, such as those involved in Creutzfeldt-Jakob disease. In variant Creutzfeldt-Jakob disease, prion replication is thought to occur first in the lymphoreticular system and then spread into the brain. We have, therefore, examined the susceptibility of a human tonsil-derived follicular dendritic cell-like cell line (HK) to prion infection. HK cells were found to display a readily detectable, time-dependent increase in cell-associated abnormal prion protein (PrP(TSE)) when exposed to medium spiked with Creutzfeldt-Jakob disease brain homogenate, resulting in a coarse granular perinuclear PrP(TSE) staining pattern. Despite their high level of cellular prion protein expression, HK cells failed to support infection, as judged by longer term maintenance of PrP(TSE) accumulation. Colocalization studies revealed that exposure of HK cells to brain homogenate resulted in increased numbers of detectable lysosomes and that these structures immunostained intensely for PrP(TSE) after exposure to Creutzfeldt-Jakob disease brain homogenate. Our data suggest that human follicular dendritic-like cells and perhaps other human cell types are able to avoid prion infection by efficient lysosomal degradation of PrP(TSE). PMID:24183781

  4. Cutting edge: circulating plasmablasts induce the differentiation of human T follicular helper cells via IL-6 production.

    PubMed

    Chavele, Konstantia-Maria; Merry, Eve; Ehrenstein, Michael R

    2015-03-15

    B cells require CD4(+) T follicular helper (Tfh) cells to progress through the germinal center and provide protective Ab responses. In this article, we reveal a reciprocal interaction whereby circulating human plasmablasts are potent inducers of the Tfh cell-differentiation program, including the expression of their key transcription factor Bcl-6. The markedly increased propensity of plasmablasts, compared with naive B cells, to induce Tfh cell differentiation was due to their increased production of IL-6. Specific targeting of IL-6 using tocilizumab therapy in patients with rheumatoid arthritis led to a significant reduction in circulating Tfh cell numbers and IL-21 production, which was correlated with reduced plasmablast formation. Our data uncover a positive-feedback loop between circulating plasmablasts and Tfh cells that could sustain autoimmunity and spread Ab-driven inflammation to unaffected sites; this represents an important therapeutic target, as well as reveals a novel mechanism of action for tocilizumab. PMID:25681343

  5. The effect of a negative energy balance status on β-carotene availability in serum and follicular fluid of nonlactating dairy cows.

    PubMed

    De Bie, J; Langbeen, A; Verlaet, A A J; Florizoone, F; Immig, I; Hermans, N; Fransen, E; Bols, P E J; Leroy, J L M R

    2016-07-01

    Maternal metabolic pressure due to a cow's negative energy balance (NEB) has a negative effect on oocyte quality as a result of increased oxidative stress. In this study, we hypothesized that a NEB status may negatively affect the availability of β-carotene (bC, an antioxidant) in the micro-environment of the oocyte or follicular fluid (FF) and that daily bC supplementation can increase bC availability. We aimed to (1) determine the effect of a nutritionally induced NEB on bC concentrations in serum and FF as well as on the presence of bC metabolites, oxidative stress levels, and follicular growth in a nonlactating dairy cow model, and (2) investigate how this effect could be altered by dietary bC supplementation. Six multiparous nonlactating Holstein Friesian cows were subjected to 4 consecutive dietary treatments, 28 d each: (1) 1.2 × maintenance (M) or positive energy balance (PEB) without bC supplement (PEB-bC), (2) 1.2 × M with daily supplement of 2,000mg of bC comparable to the level of bC intake at grazing (PEB+bC), (3) 0.6 × M with 2,000mg of bC (NEB+bC), and (4) 0.6 × M (NEB-bC). At the end of each treatment, estrous cycles were synchronized and blood and FF of the largest follicle were sampled and analyzed for bC, retinol, α-tocopherol, free fatty acids, estradiol, and progesterone. Serum cholesterol, triglycerides, urea, insulin growth factor 1, growth hormone, total antioxidant status (TAS), and red blood cell glutathione (GSH) concentrations were determined as well. All cows lost body weight during both energy restriction periods and showed increased serum free fatty acid concentrations, illustrating a NEB. A dietary induced NEB reduced FF bC, but not plasma bC or plasma and FF retinol concentrations. However, bC and retinol concentrations drastically increased in both fluid compartments after bC supplementation. Follicular diameter was increased in supplemented PEB cows. Energy restriction reduced the TAS and red blood cell GSH, whereas daily b

  6. A study relating the composition of follicular fluid and blood plasma from individual Holstein dairy cows to the in vitro developmental competence of pooled abattoir-derived oocytes.

    PubMed

    Sutton-McDowall, Melanie L; Yelland, Robert; MacMillan, Keith L; Robker, Rebecca L; Thompson, Jeremy G

    2014-07-01

    The fertility of high-performance (high milk yield) dairy breeds such as the Holstein within the Australian dairy herd has been on the decline for the past two decades. The 12-month calving interval for pasture-based farming practices results in oocyte maturation coinciding with peak lactation, periods of negative energy balance, and energy partitioning for lactation, causing energy deficiency in some organ systems, including the reproductive system. Oocyte developmental competence (the ability to undergo successful fertilization, embryo development, and establishment of pregnancy) is intrinsically linked with the composition of follicular fluid (FF). The aim of this study was to determine if there was a relationship between the fat and carbohydrate levels in plasma and FF and the ability to support in vitro oocyte maturation (IVM). Plasma and FF were collected in vivo from eight Holstein cows between 52 and 151 days post-partum. Plasma glucose trended (P = 0.072) higher and triglyceride levels were significantly higher than in FF (P < 0.05), but there were no relationships between FF and plasma composition. Glucose FF concentration was negatively related to follicular lactate and nonesterified fatty acid (NEFA) levels and days post-partum. Conversely, FF triglyceride concentrations were positively related to FF NEFA levels and negatively related to milk fat and protein composition. Abattoir-derived cumulus-oocyte complexes were cultured in either 50% FF (FF-IVM) or 50% plasma (plasma-IVM), with on-time embryo development then assessed. Although there were no differences between animals, the blastocyst rates after FF-IVM were negatively related to plasma glucose and days post-partum and positively related to body condition score and plasma NEFA levels. In comparison to the previous studies, total NEFA levels in FF were not related to animal parameters and did not influence oocyte developmental competence in vitro. Results from this study suggest that days

  7. Cell-free DNA in Human Follicular Microenvironment: New Prognostic Biomarker to Predict in vitro Fertilization Outcomes

    PubMed Central

    Mullet, Tiffany; Molinari, Nicolas; Vincens, Claire; Anahory, Tal; Hamamah, Samir

    2015-01-01

    Cell-free DNA (cfDNA) fragments, detected in blood and in other biological fluids, are released from apoptotic and/or necrotic cells. CfDNA is currently used as biomarker for the detection of many diseases such as some cancers and gynecological and obstetrics disorders. In this study, we investigated if cfDNA levels in follicular fluid (FF) samples from in vitro fertilization (IVF) patients, could be related to their ovarian reserve status, controlled ovarian stimulation (COS) protocols and IVF outcomes. Therefore, 117 FF samples were collected from women (n = 117) undergoing IVF/Intra-cytoplasmic sperm injection (ICSI) procedure and cfDNA concentration was quantified by ALU-quantitative PCR. We found that cfDNA level was significantly higher in FF samples from patients with ovarian reserve disorders (low functional ovarian reserve or polycystic ovary syndrome) than from patients with normal ovarian reserve (2.7 ± 2.7 ng/μl versus 1.7 ± 2.3 ng/μl, respectively, p = 0.03). Likewise, FF cfDNA levels were significant more elevated in women who received long ovarian stimulation (> 10 days) or high total dose of gonadotropins (≥ 3000 IU/l) than in women who received short stimulation duration (7–10 days) or total dose of gonadotropins < 3000 IU/l (2.4 ± 2.8 ng/μl versus 1.5 ± 1.9 ng/μl, p = 0.008; 2.2 ± 2.3 ng/μl versus 1.5 ± 2.1 ng/μl, p = 0.01, respectively). Finally, FF cfDNA level was an independent and significant predictive factor for pregnancy outcome (adjusted odds ratio = 0.69 [0.5; 0.96], p = 0.03). In multivariate analysis, the Receiving Operator Curve (ROC) analysis showed that the performance of FF cfDNA in predicting clinical pregnancy reached 0.73 [0.66–0.87] with 88% specificity and 60% sensitivity. CfDNA might constitute a promising biomarker of follicular micro-environment quality which could be used to predict IVF prognosis and to enhance female infertility management. PMID:26288130

  8. Comparison of follicular fluid and serum levels of Inhibin A and Inhibin B with calculated indices used as predictive markers of ovarian hyperstimulation syndrome in IVF patients

    PubMed Central

    Moos, Jiri; Rezabek, Karel; Filova, Vanda; Moosova, Martina; Pavelkova, Jana; Peknicova, Jana

    2009-01-01

    Background Ovarian Hyperstimulation Syndrome (OHSS) is a severe health complication observed in some patients undergoing hormonal stimulation during IVF. Presence of OHSS is often associated with a high count of growing follicles responding to FSH hyperstimulation. However, the number of responding follicles may not be sufficient enough to predict the onset and severity of OHSS. The aim of this study was to find whether follicular fluid (FF) and serum concentrations of Inhibin A and Inhibin B in patients undergoing IVF treatment may serve as a predictor of OHSS status independent of the growing follicles count. Methods Serum and follicular fluid of fifty-three women undertaking the IVF program were separated into four groups according to their OHSS status and growing follicles count and analyzed for serum and FF concentrations of Inhibin A and Inhibin B. The resulting data were combined with clinical and demographic data to calculate indices independent of the growing follicles count. Results Serum Inhibin A and Inhibin B concentrations showed no significant difference between the severe OHSS group and the control group without OHSS. Moreover, the serum concentrations of Inhibin A and Inhibin B were strongly correlated with the growing follicles count. Their concentrations in the high responders group (>18 follicles) were significantly higher (p < 0.00001, p < 0.0001) when compared with normal and low responders (<18 follicles). To suppress the dependence on the growing follicle count, three indices were constructed and calculated. The best association with OHSS status and independence of the growing follicle count was achieved by using the Inhibin B TFF/SBM index calculated as follows: [concentration in FF] × [growing follicle count]/[concentration in serum] × [body mass]. The Inhibin B TFF/SBM index showed a clear difference (p = 0,00433) between the group with severe OHSS and the control group, while showing no apparent correlation with the growing follicle

  9. Anti-Müllerian Hormone Levels in the Follicular Fluid of the Preovulatory Follicle: A Predictor for Oocyte Fertilization and Quality of Embryo

    PubMed Central

    2014-01-01

    This prospective study investigated the relationship between anti-Müllerian hormone (AMH) level in the follicular fluid (FF) and the quality of the oocyte and embryo. A total of 65 FF samples from 54 women were included in this study. FF was collected from the largest preovulatory follicle sized≥20 mm of mean diameter from each ovary. Samples were divided into 3 groups according to the FF AMH levels: below the 33th percentile (low group, FF AMH<2.1 ng/mL, n=21), between the 33th and the 67th percentile (intermediate group, FF AMH=2.1-3.6 ng/mL, n=22), and above the 67th percentile (high group, FF AMH>3.6 ng/mL, n=22). The quality of the ensuing oocytes and embryos was evaluated by fertilization rate and embryo score. FF AMH levels correlated positively with the matched embryo score on day 3 after fertilization (r=0.331, P=0.015). The normal fertilization rate was significantly lower in the low group than in the intermediate group (61.9% vs. 95.5% vs. 77.3%, respectively, P=0.028). Our results suggest that the FF AMH level could be a predictor of the ensuing oocyte and embryo quality. Graphical Abstract PMID:25246746

  10. Anti-Mullerian hormone (AMH) concentration in follicular fluid and mRNA expression of AMH receptor type II and LH receptor in granulosa cells as predictive markers of good buffalo (Bubalus bubalis) donors.

    PubMed

    Liang, Aixin; Salzano, Angela; D'Esposito, Maurizio; Comin, Antonella; Montillo, Marta; Yang, Liguo; Campanile, Giuseppe; Gasparrini, Bianca

    2016-09-01

    High individual variability in follicular recruitment and hence in the number of embryos produced is a major factor limiting the application of reproductive technologies in buffalo. Therefore, the identification of reliable markers to select embryo donors is critical to enroll buffaloes in embryo production programs. Better understanding of factors involved in follicular growth is also necessary to improve the response to superovulation in this species. The aim of this work was thus to determine the anti-Mullerian hormone (AMH) concentration in follicular fluid (FF) recovered from different size follicles and evaluate the mRNA expression profiles of development-related (AMHR2, CYP19A1, FSHR, and LHR) and apoptosis-related genes (TP53INP1 and CASP3) in the corresponding granulosa cells (GCs) in buffalo. Another objective was to evaluate whether the AMH concentration in FF and gene expression of GCs is associated with the antral follicular count. Ovaries were collected at the slaughterhouse, and all follicles were counted and classified as small (3-5 mm), medium (5-8 mm), and large (>8 mm). Follicular fluid was recovered for AMH determination, and the mRNA expression of AMHR2, FSHR, LHR, CYP19A1, TP53INP1, and CASP3 was analyzed in GCs. The AMH concentration in FF decreased (P < 0.01) at increasing follicular diameter. The mRNA expression of AMHR2 and FSHR was higher (P < 0.05) in small follicles, whereas that of LHR and CYP19A1 was higher (P < 0.05) in large follicles. The intrafollicular AMH concentration was positively correlated with the antral follicular count (r = 0.31; P < 0.05). Interestingly, good donors (≥12 follicles) had a higher (P < 0.05) concentration of AMH and AMHR2 levels in small follicles and higher (P < 0.05) LHR levels in large follicles than bad donors (<12 follicles). These results suggest a potential use of AMH to select buffalo donors to enroll in embryo production programs, laying the basis for further investigations

  11. Neuropilin-1 expression characterizes T follicular helper (Tfh) cells activated during B cell differentiation in human secondary lymphoid organs.

    PubMed

    Renand, Amédée; Milpied, Pierre; Rossignol, Julien; Bruneau, Julie; Lemonnier, François; Dussiot, Michael; Coulon, Séverine; Hermine, Olivier

    2013-01-01

    T follicular helper (Tfh) cells play an essential role in the development of antigen-specific B cell immunity. Tfh cells regulate the differentiation and survival of activated B cells outside and inside germinal centers (GC) of secondary lymphoid organs. They act through cognate contacts with antigen-presenting B cells, but there is no current marker to specifically identify those Tfh cells which productively interact with B cells. Here we show that neuropilin 1 (Nrp1), a cell surface receptor, is selectively expressed by a subset of Tfh cells in human secondary lymphoid organs. Nrp1 expression on Tfh cells correlates with B cell differentiation in vivo and in vitro, is transient, and can be induced upon co-culture with autologous memory B cells in a cell contact-dependent manner. Comparative analysis of ex vivo Nrp1(+) and Nrp1(-) Tfh cells reveals gene expression modulation during activation. Finally, Nrp1 is expressed by malignant Tfh-like cells in a severe case of angioimmunoblastic T-cell lymphoma (AITL) associated with elevated terminal B cell differentiation. Thus, Nrp1 is a specific marker of Tfh cells cognate activation in humans, which may prove useful as a prognostic factor and a therapeutic target in neoplastic diseases associated with Tfh cells activity. PMID:24386482

  12. Granulocyte colony-stimulating factor: A relation between serum and follicular fluid levels and in-vitro fertilization outcome in patients with polycystic ovary syndrome.

    PubMed

    Kahyaoglu, Inci; Yılmaz, Nafiye; Timur, Hakan; Inal, Hasan Ali; Erkaya, Salim

    2015-07-01

    Evidence is accumulating in the literature about the potential role of serum and follicular fluid (FF) granulocyte colony-stimulating factor (G-CSF) as a non-invasive biomarker of oocyte competence and embryo selection in in-vitro fertilization (IVF) cycles. In this study, we aimed to evaluate the effect of serum and FF G-CSF levels on IVF outcome in non-hyperandrogenic, non-obese patients with polycystic ovary syndrome (PCOS). Twenty-two patients with PCOS (Group I), and 22 patients with the etiology of male factor infertility (Group II) undergoing IVF treatment were included. Demographic features, controlled ovarian stimulation parameters, neutrophil count (NC), neutrophil/leukocyte (N/L) ratio, serum and FF G-CSF levels of the two groups were compared. Serum E2 level on the day of hCG (2982.5±171.4 vs. 2279.0±207.2 pg/mL), total number of retrieved oocytes (14.7±0.9 vs. 11.5±1.3) and mature oocytes (11.6±0.8 vs. 9.1±1.1) were significantly higher in group I when compared to group II (p<0.05). On the day of oocyte retrieval, both the mean serum (54.8±1.7 vs. 48.1±0.9 pg/mL) and FF G-CSF levels (48.8±1.4 vs. 44.1±0.5 pg/mL), NC (4.4±0.2×10(3) vs. 3.6±0.3×10(3)/μL) and N/L ratio (63.6±1.4 vs. 56.1±1.7) in group I were found to be significantly higher than group II ((p<0.05). Despite the increased levels of G-CSF both in the serum and follicular microenvironment in patients with PCOS, a relation between G-CSF and good ovarian response or clinical pregnancy rates could not be demonstrated in this study. PMID:25258001

  13. Inactivation of human interferon by body fluids

    NASA Technical Reports Server (NTRS)

    Cesario, T. C.; Mandell, A.; Tilles, J. G.

    1973-01-01

    Description of the effects of human feces, bile, saliva, serum, and cerebrospinal fluid on interferon activity. It is shown that crude interferon is inactivated by at least 50% more than with the control medium used, when incubated for 4 hr in vitro in the presence of serum, saliva, or cerebrospinal liquid, and by close to 100% when incubated with stool extract or bile.

  14. Pathways Regulating Spheroid Formation of Human Follicular Thyroid Cancer Cells under Simulated Microgravity Conditions: A Genetic Approach

    PubMed Central

    Riwaldt, Stefan; Bauer, Johann; Wehland, Markus; Slumstrup, Lasse; Kopp, Sascha; Warnke, Elisabeth; Dittrich, Anita; Magnusson, Nils E.; Pietsch, Jessica; Corydon, Thomas J.; Infanger, Manfred; Grimm, Daniela

    2016-01-01

    Microgravity induces three-dimensional (3D) growth in numerous cell types. Despite substantial efforts to clarify the underlying mechanisms for spheroid formation, the precise molecular pathways are still not known. The principal aim of this paper is to compare static 1g-control cells with spheroid forming (MCS) and spheroid non-forming (AD) thyroid cancer cells cultured in the same flask under simulated microgravity conditions. We investigated the morphology and gene expression patterns in human follicular thyroid cancer cells (UCLA RO82-W-1 cell line) after a 24 h-exposure on the Random Positioning Machine (RPM) and focused on 3D growth signaling processes. After 24 h, spheroid formation was observed in RPM-cultures together with alterations in the F-actin cytoskeleton. qPCR indicated more changes in gene expression in MCS than in AD cells. Of the 24 genes analyzed VEGFA, VEGFD, MSN, and MMP3 were upregulated in MCS compared to 1g-controls, whereas ACTB, ACTA2, KRT8, TUBB, EZR, RDX, PRKCA, CAV1, MMP9, PAI1, CTGF, MCP1 were downregulated. A pathway analysis revealed that the upregulated genes code for proteins, which promote 3D growth (angiogenesis) and prevent excessive accumulation of extracellular proteins, while genes coding for structural proteins are downregulated. Pathways regulating the strength/rigidity of cytoskeletal proteins, the amount of extracellular proteins, and 3D growth may be involved in MCS formation. PMID:27070589

  15. Metal binding components in human amniotic fluid

    SciTech Connect

    Paterson, P.G.; Zlotkin, S.H.; Sarkar, B. )

    1990-02-26

    Amniotic fluid is a potential source of both nutritionally essential and toxic metals for the fetus. As the binding pattern of these metals in amniotic fluid may be one of the determining factors in their availability to the fetus, the objective of this study was to investigate metal binding in vitro. The binding of six trace metals, Mn(II), Ni(II), Zn(II), Cu(II), Cd(II), and Fe(III), to components of human amniotic fluid was studied by Sephadex G-100 gel filtration at physiological pH, using radioisotopes as tracers and 50 mM TRIS/HCl as the elution buffer. The amniotic fluid was collected at 16-16.5 weeks gestation by amniocentesis and pooled for analysis. Extensive amounts of Fe, Cu, Zn, and Cd and small amounts of Mn and Ni were bound to high molecular weight proteins with elution patterns similar to those seen for the binding of these metals in serum. In addition, large amounts of Fe, Mn, Ni and Cd and small amounts of Zn and Cu were associated with low molecular weight component(s). The identity of these latter components is unknown, but they play an important biological role in amniotic fluid.

  16. The fluid dynamics of human birth

    NASA Astrophysics Data System (ADS)

    Lehn, Andrea; Leftwich, Megan C.

    2012-11-01

    This study investigates the fluid dynamics associated with the human birth process. Specifically, we investigate the role of the viscosity of the amniotic fluid in transferring force from the contracting uterus to the fetus during delivery. This experimental work uses an approximate uterus and dilated cervix-fabricated with liquid latex-filled with a fluid of known viscosity and an oblong solid fetus. The force required to extract the fetus is recorded for several values of amniotic viscosity. The study looks at both pull-out force values (where the fetus is pulled from outside the uterus) and push-out force values (where pressure in the experimental uterus is used to remove the fetus). In addition to the viscosity study, we also investigate the increased force required to deliver an offset fetus by tilting the major axis of the oblong fetus and repeating the pull-and push-out experiments. This study will provide knowledge about the fundamental fluid dynamic processes involved in human birth.

  17. In vivo collection of follicular fluid and granulosa cells from individual follicles of different diameters in cattle by an adapted ovum pick-up system

    PubMed Central

    2013-01-01

    Background Most studies on granulosa cell (GC) function in cattle have been performed using GC and follicular fluid (FF) samples collected from slaughterhouse ovaries. Using this approach, the follicular developmental stage and functional status are unknown and indirectly inferred, limiting data interpretation. Ultrasound-guided follicle aspiration has previously been used to recover GC or FF samples, but this was mostly carried out in large follicles or pools of small follicles, without recording the efficiency of recovery. The present study was aimed at adapting and evaluating an ovum pick-up (OPU) system for the in vivo recovery of FF and GC from individual follicles of different diameters. Methods In the first trial, the losses of fluid inside the tubing system were calculated using a conventional or an adapted-OPU system. Blood plasma volumes equivalent to the amount of FF in follicles of different diameters were aspirated using a conventional OPU Teflon circuit. The OPU system was then adapted by connecting 0.25 mL straws to the circuit. A second trial evaluated the efficiency of FF recovery in vivo. Follicles ranging from 4.0 to 16.8 mm in diameter were aspirated individually using the conventional or adapted-OPU systems. A third trial assessed the in vivo recovery of GC and the subsequent amount of RNA obtained from the follicles of different diameters from Holstein and Gir cattle. Results In Trial I, the plasma recovery efficiency was similar (P > 0.05) for the volumes expected for 12 and 10 mm follicles, but decreased (P < 0.05) for smaller follicles (45.7+/−4.0%, 12.4+/−4.3% and 0.0+/−0.0% for 8, 6, and 4 mm follicles, respectively). Using the adaptation, the losses intrinsic to the aspiration system were similar for all follicle diameters. In Trial II, the expected and recovered volumes of FF were correlated (r = 0.89) and the efficiency of recovery was similar among follicles <12 mm, while larger follicles had a progressive increase

  18. Different patterns of 5{alpha}-reductase expression, cellular distribution, and testosterone metabolism in human follicular dermal papilla cells

    SciTech Connect

    Liu, Shicheng Yamauchi, Hitoshi

    2008-04-18

    Androgens regulate hair growth, and 5{alpha}-reductase (5{alpha}R) plays a pivotal role in the action of androgens on target organs. To clarify the molecular mechanisms responsible for controlling hair growth, the present study presents evidence that the human follicular dermal papilla cells (DPCs) from either beard (bDPCs) or scalp hair (sDPCs) possess endogenous 5{alpha}R activity. Real-time RT-PCR revealed that the highest level of 5{alpha}R1 mRNA was found in bDPCs, followed by sDPCs, and a low but detectable level of 5{alpha}R1 mRNA was observed in fibroblasts. Minimally detectable levels of 5{alpha}R2 mRNA were found in all three cell types. A weak band at 26 kDa corresponding to the human 5{alpha}R1 protein was detected by Western blot in both DPCs, but not in fibroblasts. Immuonofluorescence analysis confirmed that 5{alpha}R1 was localized to the cytoplasm rather than in the nuclei in both DPCs Furthermore, a 5{alpha}R assay using [{sup 14}C]testosterone labeling in intact cells revealed that testosterone was transformed primarily into androstenedione, and in small amounts, into DHT. Our results demonstrate that the 5{alpha}R activities of either bDPCs or sDPCs are stronger than that of dermal fibroblasts, despite the fact that the major steroidogenic activity is attributed to 17{beta}-HSD rather than 5{alpha}R among the three cell types. The 5{alpha}R1 inhibitor MK386 exhibited a more potent inhibitory effect on 5{alpha}R activity than finasteride (5{alpha}R2 inhibitor) in bDPCs.

  19. Analysis of follicular fluid retinoids in women undergoing in vitro fertilization: retinoic acid influences embryo quality and is reduced in women with endometriosis.

    PubMed

    Pauli, Samuel A; Session, Donna R; Shang, Weirong; Easley, Kirk; Wieser, Friedrich; Taylor, Robert N; Pierzchalski, Keely; Napoli, Joseph L; Kane, Maureen A; Sidell, Neil

    2013-09-01

    Retinol (ROL) and its biologically active metabolite, all-trans retinoic acid (ATRA), are essential for a number of reproductive processes. However, there is a paucity of information regarding their roles in ovarian folliculogenesis, oocyte maturation, and early embryogenesis. The objectives of this study were to quantify and compare peripheral plasma (PP) and follicular fluid (FF) retinoid levels, including ATRA in women undergoing in vitro fertilization (IVF) and to investigate the relationship between retinoid levels and embryo quality. Retinoid levels were evaluated in PP and FF from 79 women undergoing IVF at the time of oocyte retrieval and corresponding embryo quality assessed on a daily basis after retrieval for 3 days until uterine transfer. Analysis compared the retinoid levels with day 3 embryo grades and between endometriosis versus control patients. Results demonstrated distinctive levels of retinoid metabolites and isomers in FF versus PP. There was a significantly larger percentage of high-quality grade I embryos derived from the largest versus smallest follicles. An increase in follicle size also correlated with a >50% increase in FF ROL and ATRA concentrations. Independent of follicle size, FF yielding grade I versus nongrade I embryos showed higher mean levels of ATRA but not ROL. In a nested case-control analysis, control participants had 50% higher mean levels of ATRA in their FF and PP than women with endometriosis. These findings strongly support the proposition that ATRA plays a fundamental role in oocyte development and quality, and that reduced ATRA synthesis may contribute to decreased fecundity of participants with endometriosis. PMID:23427183

  20. N-Acetyl-Cysteine and l-Carnitine Prevent Meiotic Oocyte Damage Induced by Follicular Fluid From Infertile Women With Mild Endometriosis.

    PubMed

    Giorgi, Vanessa S I; Da Broi, Michele G; Paz, Claudia C P; Ferriani, Rui A; Navarro, Paula A

    2016-03-01

    This study evaluated the potential protective effect of the antioxidants, l-carnitine (LC) and N-acetyl-cysteine (NAC), in preventing meiotic oocyte damage induced by follicular fluid (FF) from infertile women with mild endometriosis (ME). We performed an experimental study. The FF samples were obtained from 22 infertile women undergoing stimulated cycles for intracytoplasmic sperm injection (11 with ME and 11 without endometriosis). Immature bovine oocytes were submitted to in vitro maturation (IVM) divided into 9 groups: no-FF (No-FF); with FF from control (CFF) or ME (EFF) groups; and with LC (C + LC and E + LC), NAC (C + NAC and E + NAC), or both antioxidants (C + 2Ao and E + 2Ao). After IVM, oocytes were immunostained for visualization of microtubules and chromatin by confocal microscopy. The percentage of meiotically normal metaphase II (MII) oocytes was significantly lower in the EFF group (51.35%) compared to No-FF (86.36%) and CFF (83.52%) groups. The E + NAC (62.22%), E + LC (80.61%), and E + 2Ao (61.40%) groups showed higher percentage of normal MII than EFF group. The E + LC group showed higher percentage of normal MII than E + NAC and E + 2Ao groups and a similar percentage to No-FF and CFF groups. Therefore, FF from infertile women with ME causes meiotic abnormalities in bovine oocytes, and, for the first time, we demonstrated that the use of NAC and LC prevents these damages. Our findings elucidate part of the pathogenic mechanisms involved in infertility associated with ME and open perspectives for further studies investigating whether the use of LC could improve the natural fertility and/or the results of in vitro fertilization of women with ME. PMID:26342050

  1. Restorative effect of hair follicular dermal cells on injured human hair follicles in a mouse model.

    PubMed

    Yamao, Mikaru; Inamatsu, Mutsumi; Okada, Taro; Ogawa, Yuko; Ishida, Yuji; Tateno, Chise; Yoshizato, Katsutoshi

    2015-03-01

    No model is available for examining whether in vivo-damaged human hair follicles (hu-HFs) are rescued by transplanting cultured hu-HF dermal cells (dermal papilla and dermal sheath cells). Such a model might be valuable for examining whether in vivo-damaged hu-HFs such as miniaturized hu-HFs in androgenic alopecia are improvable by auto-transplanting hu-HF dermal cells. In this study, we first developed mice with humanized skin composed of hu-keratinocytes and hu-dermal fibroblasts. Then, a 'humanized scalp model mouse' was generated by transplanting hu-scalp HFs into the humanized skin. To demonstrate the usability of the model, the lower halves of the hu-HFs in the model were amputated in situ, and cultured hu-HF dermal cells were injected around the amputated area. The results demonstrated that the transplanted cells contributed to the restoration of the damaged HFs. This model could be used to explore clinically effective technologies for hair restoration therapy by autologous cell transplantation. PMID:25557326

  2. 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)

    PubMed Central

    2014-01-01

    Background 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. Methods 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. Results 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. Conclusions 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. PMID:25078409

  3. Nature and importance of follicular lymphoma precursors

    PubMed Central

    Mamessier, Emilie; Broussais-Guillaumot, Florence; Chetaille, Bruno; Bouabdallah, Reda; Xerri, Luc; Jaffe, Elaine S.; Nadel, Bertrand

    2014-01-01

    It is now widely recognized that cancer development is a protracted process requiring the stepwise acquisition of multiple oncogenic events. In humans, this process can take decades, if not a lifetime, blurring the notion of ‘healthy’ individuals. Follicular lymphoma exemplifies this multistep pathway of oncogenesis. In recent years, variants of follicular lymphoma have been recognized that appear to represent clonal B-cell expansions at an early stage of follicular lymphoma lymphomagenesis. These include follicular lymphoma in situ, duodenal follicular lymphoma, partial involvement by follicular lymphoma, and in the blood circulating follicular lymphoma-like B cells. Recent genetic studies have identified similarities and differences between the early lesions and overt follicular lymphoma, providing important information for understanding their biological evolution. The data indicate that there is already genomic instability at these early stages, even in instances with a low risk for clinical progression. The overexpression of BCL2 in t(14;18)-positive B cells puts them at risk for subsequent genetic aberrations when they re-enter the germinal center and are exposed to the influences of activation-induced cytidine deaminase and somatic hypermutations. The emerging data provide a rationale for clinical management and, in the future, may identify genetic risk factors that warrant early therapeutic intervention. PMID:24790058

  4. Effects of tamoxifen and somatostatin analogue on growth of human medullary, follicular, and papillary thyroid carcinoma cell lines: tissue culture and nude mouse xenograft studies.

    PubMed

    Weber, C J; Marvin, M; Krekun, S; Koschitzky, T; Karp, F; Benson, M; Feind, C R

    1990-12-01

    The knowledge that (1) the normal thyroid contains somatostatin, (2) polypeptide growth factors influence thyroid cell function, and (3) thyroid cells contain steroid hormone receptors prompted us to add somatostatin analogue No. 201-995 (SMS) (5 ng/ml) and/or tamoxifen citrate (TAM) (5 mumol/L) to 7-day monolayer cultures (50,000 cells/well) of three separate human thyroid carcinoma cell lines: DR081 (medullary), WR082 (follicular), and NPA'87 (papillary). Results, tabulated as cell numbers/well (X10(5) on day 7, revealed that TAM inhibited growth of medullary and follicular cells and that TAM plus SMS inhibited growth of papillary cells. In vivo studies of subcutaneous tumor cell xenografts in nude mice have documented that TAM (5 mg subcutaneous pellet) significantly inhibits the growth of medullary implants. Flow cytometric DNA studies of medullary cell cultures demonstrated a reduced G2 + M phase with TAM treatment. For papillary cell implants, TAM plus SMS (5 micrograms subcutaneously, twice daily) did not suppress tumor growth. All three cell lines were negative for estrogen receptor; addition of estradiol (5 ng/ml) to medullary cell cultures neither stimulated replication nor reversed the inhibitory effects of TAM in vitro. We conclude that (1) TAM slowed the growth of a cell line of human medullary carcinoma, both in vitro and in vivo; (2) this effect was not reversed by estradiol; (3) TAM plus SMS inhibited replication of a papillary carcinoma cell line in vitro, but not in vivo; and (4) TAM alone and TAM plus SMS inhibited replication of cultures of a human follicular thyroid carcinoma cell line. TAM and SMS may be useful in treatment of some human thyroid carcinomas. PMID:1978945

  5. Nitric oxide concentrations, estradiol-17β progesterone ratio in follicular fluid, and COC quality with respect to perifollicular blood flow in cows.

    PubMed

    Pancarci, S M; Ari, U Ç; Atakisi, O; Güngör, O; Ciğremiş, Y; Bollwein, H

    2012-01-01

    The objectives were to investigate relationships among concentrations of nitric oxide (NO), estradiol 17 beta (E2), and progesterone (P4) in follicular fluids (FF), and quality of cumulus oocyte complexes (COCs) with respect to perifollicular blood flow (FBF). In Experiment I, follicles (138) were classified according to the presence or absence of FBF (assessed with transvaginal Doppler ultrasonography) and diameter of follicles (small, 2-4 mm; medium, 5-8 mm; and large, ≥9 mm). Concentrations of NO in FF did not differ significantly among these size categories. However, NO concentrations in FF with FBF (54.4 ± 7.4 μmol/l) were higher (P<0.05) than in those without FBF (36.6 ± 4.1 μmol/l). There was a positive correlation (r=0.30, P<0.05) between NO concentrations and the E2:P4 in FF. Rate of E2 active (E2:P4 ≥ 1) follicles were numerically 1.2 (0.8-1.8) times higher in follicles with FBF (38.1%) compared to those without FBF (25.0%). Moreover, rates of E2 active follicles were 6.1 (0.7-55.2) and 1.3 (0.1-17.3) times higher (P<0.06) in large (43.3%) and medium (14.3%) compared to small follicles (11.1%), respectively. In Experiment II, quality of COCs from 2 to 8 mm follicles, obtained by transvaginal ovum pick up (OPU), was investigated with respect to FBF. Odds ratio to obtain higher quality COCs from follicles with FBF (47.1%) was 3.3 (1.1-9.6) fold higher (P<0.05) compared to those from follicles without FBF (14.6%). In conclusion, E2:P4, and NO concentrations in FF, as well as FBF, could be used to determine the functionality of ovarian follicles in cows. Moreover, determination of FBF could be useful to predict quality of COCs in cattle. PMID:22281101

  6. Fatty acid composition of the follicular fluid of normal weight, overweight and obese women undergoing assisted reproductive treatment: a descriptive cross-sectional study

    PubMed Central

    2014-01-01

    Background It has been well documented that the maturing oocyte is very vulnerable to changes in its micro-environment, the follicular fluid (FF). Recent research has focused on different components within this FF, like hormones, growth factors and metabolites, and how their concentrations are altered by diet and the metabolic health of the mother. It has been proposed that fatty acids (FAs) are potential factors that influence oocyte maturation and subsequent embryo development. However, a thorough study of the specific FF FA composition per lipid fraction and how this may be affected by BMI is currently lacking. Therefore, we investigated the BMI-related concentration of FAs in the phospholipid (PL), cholesteryl-ester (CHE), triglyceride (TG) and non-esterified (NE) lipid fraction in the FF of women undergoing assisted reproductive treatment (ART). Methods In this descriptive cross-sectional study, the FF of normal weight (18.5 ≤ BMI < 25.0 kg/m(2), n = 10), overweight (25.0 ≤ BMI < 30.0 kg/m(2), n = 10) and obese (BMI ≥ 30.0 kg/m(2), n = 10) women, undergoing ART, was sampled and analyzed for 23 specific FAs in the PL, CHE, TG and NEFA fraction, using a gas chromatographic analysis method. Differences between BMI-groups were studied by means of univariate general linear models and post hoc Sheffé tests. Results Total FA concentrations in the PL and CHE fraction did not differ between BMI groups. Total TG concentrations tended to differ and total NEFA concentrations differed significantly between BMI groups. Interestingly, 42% and 34% of the total FAs was esterified in the PL and CHE fraction, respectively, while only 10% were present in both the TG and NEFA fraction. Only few individual FA concentrations differed in the PL, CHE and TG fraction between BMI groups, whereas abundant BMI-related differences were found in the NEFA fraction. Conclusions Our data show that differences in BMI are associated with alterations in

  7. Diagnostic opportunities for evaluation of the exposure of dairy cows to the mycotoxins deoxynivalenol (DON) and zearalenone (ZEN): reliability of blood plasma, bile and follicular fluid as indicators.

    PubMed

    Winkler, J; Kersten, S; Meyer, U; Stinshoff, H; Locher, L; Rehage, J; Wrenzycki, C; Engelhardt, U H; Dänicke, S

    2015-10-01

    To investigate the usefulness of follicular fluid (FF) in relation to blood plasma and bile as indicators of exposure of dairy cows to ZEN, DON and their metabolites, a dose-response study was performed with 30 dairy cows. The cows, 10 in each group (named CON; FUS-50, FUS-100), received a diet with three different concentrations of Fusarium toxin-contaminated maize. Thereby, the following dietary concentration were reached: CON (0.02 mg ZEN and 0.07 mg DON, per kg dry matter, DM), FUS-50 (0.33 mg ZEN and 2.62 mg DON, per kg DM) and FUS-100 (0.66 mg ZEN and 5.24 mg DON, per kg DM). ZEN, DON and de-epoxy-DON (de-DON) were detected in FF. Based on the linear regression between toxin concentration in plasma and FF, it seems that about 50% (m = 0.5) of ZEN present in plasma is present in FF while an increase of 1 ng/ml DON or de-DON in plasma is paralleled by an increase of 1.5 ng/ml DON or 1.1 ng/ml de-DON in FF. ZEN, DON and their metabolites, except zearalenone (ZAN), were also detected in bile. Contrary to DON and de-DON, ZEN and its metabolites were accumulated in bile so that the concentration of ZEN and metabolites was much higher than for DON and de-DON. The main compound was β-zearalenol (β-ZEL). The biliary ZEN, α-zearalenol (α-ZEL) and β-ZEL concentration correlated linearly with each other with an uncertainty of <15% (r(2) ≥ 0.86), whereas the ratio between ZEN: α-ZEL: β-ZEL was about 1.5:1:11. With the help of established linear relationship between toxin intake and toxin concentration, bile could be used as diagnostic indicator to assess the exposure of cows. PMID:25556890

  8. Circulating CXCR5+CD4+ T Follicular-Like Helper Cell and Memory B Cell Responses to Human Papillomavirus Vaccines

    PubMed Central

    Matsui, Ken; Adelsberger, Joseph W.; Kemp, Troy J.; Baseler, Michael W.; Ledgerwood, Julie E.; Pinto, Ligia A.

    2015-01-01

    Through the interaction of T follicular helper (Tfh) cells and B cells, efficacious vaccines can generate high-affinity, pathogen-neutralizing antibodies, and memory B cells. Using CXCR5, CXCR3, CCR6, CCR7, PD1, and ICOS as markers, Tfh-like cells can be identified in the circulation and be classified into three functionally distinct subsets that are PD1+ICOS+, PD1+ ICOS-, or PD1-ICOS-. We used these markers to identify different subsets of CXCR5+CD4+ Tfh-like cells in response to highly immunogenic and efficacious vaccines for human papillomaviruses (HPV): Cervarix and Gardasil. In this small study, we used PBMC samples from 11 Gardasil recipients, and 8 Cervarix recipients from the Vaccine Research Center 902 Study to examine the induction of circulating Tfh-like cells and IgD-CD38HiCD27+ memory B cells by flow cytometry. PD1+ICOS+ CXCR3+CCR6-CXCR5+CD4+ (Tfh1-like) cells were induced and peaked on Day (D) 7 post-first vaccination, but not as much on D7 post-third vaccination. We also observed a trend toward increase in PD1+ICOS+ CXCR3-CCR6-CXCR5+CD4+ (Tfh2-like) cells for both vaccines, and PD1+ICOS+ CXCR3-CCR6+CXCR5+CD4+ (Tfh17-like) subset was induced by Cervarix post-first vaccination. There were also minimal changes in the other cellular subsets. In addition, Cervarix recipients had more memory B cells post-first vaccination than did Gardasil recipients at D14 and D30. We found frequencies of memory B cells at D30 correlated with anti-HPV16 and 18 antibody titers from D30, and the induction levels of memory B cells at D30 and PD1+ICOS+Tfh1-like cells at D7 post-first vaccination correlated for Cervarix. Our study showed that induction of circulating CXCR5+CD4+ Tfh-like subsets can be detected following immunization with HPV vaccines, and potentially be useful as a marker of immunogenicity of vaccines. However, further investigations should be extended to different cohorts with larger sample size to better understand the functions of these T cells, as well as

  9. Follicular disorders of the face.

    PubMed

    Sardana, Kabir

    2014-01-01

    Follicular disorders are possibly a commonly seen, though rarely reported, group of disorders, which are localized around follicles and appendages and clinically present as small papules. Classically, clinicopathological follicular plugging is seen in the "true" follicular disorders, which are appropriately referred to as follicular keratosis. Another subtype is folliculitis (infectious or noninfectious) and the follicular mimickers that clinically resemble "true" follicular disorders without the follicular keratoses. Acne vulgaris has an amalgamation of all the above lesion types. Our review is aimed at describing all the follicular disorders and gives an algorithm for diagnosing the common follicular disorders. PMID:25441478

  10. Fine Structures of the Oocyte in Relation to Serum, Follicular Fluid Steroid Hormones and IGF-I in the Ovulatory-Sized Follicles in One-Humped Camel (Camelus dromedarius)

    PubMed Central

    Kafi, Mojtaba; Mesbah, Seyed Fakhroddin; Davoodian, Najmeh; Kadivar, Ali

    2014-01-01

    Background The following study was carried out to determine the ultrastructural features of the oocyte of the ovulatory-sized follicles in relation to concentrations of steroids and IGF-I in the Follicular Fluid (FF) and serum in the dromedary camel. Methods Camel follicles with a clear and healthy appearance were categorized into three classes: follicles 10 to 13.9, 14-17.9 and 18-30 mm diameter. The FF and serum samples were assayed for estradiol-17β, progesterone and IGF-I. Recovered Cumulus-Oocyte Complexes (COCs) were prepared for transmission electron microscopy. Results The mean (±SD) FF concentrations of progesterone and IGF-I was significantly (p < 0.05) higher in follicles 18 to 30 mm diameter compared to other groups of follicles. There was no difference in the mean (±SD) serum estradiol-17β, progesterone and IGF-I concentrations between camels with different ovulatory-sized follicles (p > 0.05). Oocytes from follicles 18 to 30 mm diameter (group 3) showed more advanced signs of maturation including the disappearance of the nuclear envelope, increased number of microvilli in erect position, the increase in number and size of vesicles and more even distribution of the mitochondria throughout the ooplasm. Conclusion The final stages of oocyte maturation in dromedary camel is associated with increasing progesterone and IGF-I concentrations and constant high estradiol concentration in the follicular fluid which are paralleled with well-defined ultrastructural changes in oocytes. PMID:24551433

  11. Follicular occlusion tetrad.

    PubMed

    Vasanth, Vani; Chandrashekar, Byalakere Shivanna

    2014-10-01

    Follicular occlusion tetrad is a symptom complex consisting of four conditions having a similar pathophysiology. It includes Hidradenitis suppurativa, acne conglobata, dissecting cellulitis of the scalp and pilonidal sinus. The exact pathogenesis of this group of disease is unknown but evidence suggests that they share the same pathological process initiated by follicular occlusion in apocrine gland bearing areas. Though each of these conditions is commonly encountered singly, follicular occlusion tetrad as a symptom complex has been rarely reported in the literature. PMID:25396138

  12. Follicular occlusion tetrad

    PubMed Central

    Vasanth, Vani; Chandrashekar, Byalakere Shivanna

    2014-01-01

    Follicular occlusion tetrad is a symptom complex consisting of four conditions having a similar pathophysiology. It includes Hidradenitis suppurativa, acne conglobata, dissecting cellulitis of the scalp and pilonidal sinus. The exact pathogenesis of this group of disease is unknown but evidence suggests that they share the same pathological process initiated by follicular occlusion in apocrine gland bearing areas. Though each of these conditions is commonly encountered singly, follicular occlusion tetrad as a symptom complex has been rarely reported in the literature. PMID:25396138

  13. Cholesterol side-chain cleavage cytochrome P450 and 3beta-hydroxysteroid dehydrogenase expression and the concentrations of steroid hormones in the follicular fluids of different phenotypes of healthy and atretic bovine ovarian follicles.

    PubMed

    Irving-Rodgers, Helen F; Krupa, Malgorzata; Rodgers, Raymond J

    2003-12-01

    Bovine ovarian antral follicles exhibit either one or the other of two patterns of granulosa cell death in atresia. Death can commence either from the antrum and progress toward the basal lamina (antral atresia) or the converse (basal atresia). In basal atresia, the remaining live antrally situated cells appeared to continue maturing. Beyond that, little is known about these distinct patterns of atresia. Healthy (nonatretic) follicles also exhibit either one or the other of two patterns of granulosa cell shape, follicular basal lamina ultrastructure or location of younger cells within the membrana granulosa. To examine these different phenotypes, the expression of the steroidogenic enzymes cholesterol side-chain cleavage cytochrome P450 (SCC) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) in granulosa cells and concentrations of steroid hormones in follicular fluid were measured in individual histologically classified bovine antral follicles. Healthy follicles first expressed SCC and 3beta-HSD in granulosa cells only when the follicles reached an approximate threshold of 10 mm in diameter. The pattern of expression in antral atretic follicles was the same as healthy follicles. Basal atretic follicles were all <5 mm. In these, the surviving antral granulosa cells expressed SCC and 3beta-HSD. In examining follicles of 3-5 mm, basal atretic follicles were found to have substantially elevated progesterone (P < 0.001) and decreased androstenedione and testosterone compared to healthy and antral atretic follicles. Estradiol was highest in the large healthy follicles, lower in the small healthy follicles, lower still in the antral atretic follicles, and lowest in the basal atretic follicles. Our findings have two major implications. First, the traditional method of identifying atretic follicles by measurement of steroid hormone concentrations may be less valid with small bovine follicles. Second, features of the two forms of follicular atresia are so different as to

  14. Fluid Dynamics of Human Phonation and Speech

    NASA Astrophysics Data System (ADS)

    Mittal, Rajat; Erath, Byron D.; Plesniak, Michael W.

    2013-01-01

    This article presents a review of the fluid dynamics, flow-structure interactions, and acoustics associated with human phonation and speech. Our voice is produced through the process of phonation in the larynx, and an improved understanding of the underlying physics of this process is essential to advancing the treatment of voice disorders. Insights into the physics of phonation and speech can also contribute to improved vocal training and the development of new speech compression and synthesis schemes. This article introduces the key biomechanical features of the laryngeal physiology, reviews the basic principles of voice production, and summarizes the progress made over the past half-century in understanding the flow physics of phonation and speech. Laryngeal pathologies, which significantly enhance the complexity of phonatory dynamics, are discussed. After a thorough examination of the state of the art in computational modeling and experimental investigations of phonatory biomechanics, we present a synopsis of the pacing issues in this arena and an outlook for research in this fascinating subject.

  15. Metabolism of vasopressin by human amnion and amniotic fluid

    SciTech Connect

    Claybaugh, J.R.; Uyehara, C.F.T.; Sato, A.K.; Letterie, G.S. )

    1990-02-26

    The authors previously have shown that vasopressin (VP) is metabolized by fetal guinea pig amnion and amniotic fluid into 2 distinct metabolites: M1, produced by amnionic membrane, and M2, produced in amniotic fluid. In this study the authors have examined whether VP can also be metabolized by human amnionic membrane and amniotic fluid collected at term. Tritiated VP (250 pg) was incubated in vitro with either amnionic membrane or amniotic fluid for 2 hours at 37{degrees}C, followed by fractionation by HPLC. After incubation with amnionic membrane, tritiated VP was metabolized to Ml with no production of M2, similar to previous findings with guinea pig amnion. Human amniotic fluid, however, produced both M1 and M2 metabolites, unlike guinea pig amniotic fluid which metabolized VP to only M2 with no M1 production. No other metabolic products other than M1 or M2 were isolated. Also, like the M2 produced in guinea pig amniotic fluid, the M2 metabolite in human amniotic fluid comigrates on HPLC with desglycinamide VP. Thus, metabolic components in the human amniotic sac appear to be similar to those present in the guinea pig. These results indicate that the amniotic compartment may be an important clearance site for fetal VP in humans.

  16. Single-molecule imaging of hyaluronan in human synovial fluid

    NASA Astrophysics Data System (ADS)

    Kappler, Joachim; Kaminski, Tim P.; Gieselmann, Volkmar; Kubitscheck, Ulrich; Jerosch, Jörg

    2010-11-01

    Human synovial fluid contains a high concentration of hyaluronan, a high molecular weight glycosaminoglycan that provides viscoelasticity and contributes to joint lubrication. In osteoarthritis synovial fluid, the concentration and molecular weight of hyaluronan decrease, thus impairing shock absorption and lubrication. Consistently, substitution of hyaluronan (viscosupplementation) is a widely used treatment for osteoarthritis. So far, the organization and dynamics of hyaluronan in native human synovial fluid and its action mechanism in viscosupplementation are poorly characterized at the molecular level. Here, we introduce highly sensitive single molecule microscopy to analyze the conformation and interactions of fluorescently labeled hyaluronan molecules in native human synovial fluid. Our findings are consistent with a random coil conformation of hyaluronan in human synovial fluid, and point to specific interactions of hyaluronan molecules with the synovial fluid matrix. Furthermore, single molecule microscopy is capable of detecting the breakdown of the synovial fluid matrix in osteoarthritis. Thus, single molecule microscopy is a useful new method to probe the structure of human synovial fluid and its changes in disease states like osteoarthritis.

  17. Benign follicular tumors*

    PubMed Central

    Tellechea, Oscar; Cardoso, José Carlos; Reis, José Pedro; Ramos, Leonor; Gameiro, Ana Rita; Coutinho, Inês; Baptista, António Poiares

    2015-01-01

    Benign follicular tumors comprise a large and heterogeneous group of neoplasms that share a common histogenesis and display morphological features resembling one or several portions of the normal hair follicle, or recapitulate part of its embryological development. Most cases present it as clinically nondescript single lesions and essentially of dermatological relevance. Occasionally, however, these lesions be multiple and represent a cutaneous marker of complex syndromes associated with an increased risk of visceral neoplasms. In this article, the authors present the microscopic structure of the normal hair follicle as a basis to understand the type and level of differentiation of the various follicular tumors. The main clinicopathological features and differential diagnosis of benign follicular tumors are then discussed, including dilated pore of Winer, pilar sheath acanthoma, trichoadenoma, trichilemmoma, infundibuloma, proliferating trichilemmal cyst/tumor, trichoblastoma and its variants, pilomatricoma, trichodiscoma/fibrofolliculoma, neurofollicular hamartoma and trichofolliculoma. In addition, the main syndromes presenting with multiple follicular tumors are also discussed, namely Cowden, Birt-Hogg-Dubé, Rombo and Bazex-Dupré-Christol syndromes, as well as multiple tumors of follicular infundibulum (infundibulomatosis) and multiple trichoepitheliomas. Although the diagnosis of follicular tumors relies on histological examination, we highlight the importance of their knowledge for the clinician, especially when in presence of patients with multiple lesions that may be the cutaneous marker of a cancer-prone syndrome. The dermatologist is therefore in a privileged position to recognize these lesions, which is extremely important to provide further propedeutic, appropriate referral and genetic counseling for these patients. PMID:26734858

  18. Mutagenesis assays of human amniotic fluid

    SciTech Connect

    Everson, R.B.; Milne, K.L.; Warbuton, D.; McClamrock, H.D.; Buchanan, P.D.

    1985-01-01

    Extracts of amniocentesis samples from 144 women were tested for the presence of mutagenic substances using tester strain TA1538 in the Ames Salmonella/mammalian-microsome mutagenicity test. Because the volume of amniotic fluid in these samples was limited (generally less than 10 ml), the authors investigated modifications of this mutagenesis assay that could increase its ability to detect effects from small quantities of test material. Using mutagenicity in samples of urine from smokers as a model, it appeared that improved ability to detect small amounts of mutagen could be obtained by reducing volumes of media and reagents while keeping the amount of test sample constant. Tests of amniotic fluid extracts by this modified procedure showed small increases in revertants, about 50% above dimethylsulfoxide solvent control values. The increases suggest the presence of small amounts of mutagenic material in many of the amniotic fluid samples. At the doses employed, mutagenic activity in these samples was not associated with maternal smoking.

  19. Proteomic analysis of human osteoarthritis synovial fluid

    PubMed Central

    2014-01-01

    Background Osteoarthritis is a chronic musculoskeletal disorder characterized mainly by progressive degradation of the hyaline cartilage. Patients with osteoarthritis often postpone seeking medical help, which results in the diagnosis being made at an advanced stage of cartilage destruction. Sustained efforts are needed to identify specific markers that might help in early diagnosis, monitoring disease progression and in improving therapeutic outcomes. We employed a multipronged proteomic approach, which included multiple fractionation strategies followed by high resolution mass spectrometry analysis to explore the proteome of synovial fluid obtained from osteoarthritis patients. In addition to the total proteome, we also enriched glycoproteins from synovial fluid using lectin affinity chromatography. Results We identified 677 proteins from synovial fluid of patients with osteoarthritis of which 545 proteins have not been previously reported. These novel proteins included ADAM-like decysin 1 (ADAMDEC1), alanyl (membrane) aminopeptidase (ANPEP), CD84, fibulin 1 (FBLN1), matrix remodelling associated 5 (MXRA5), secreted phosphoprotein 2 (SPP2) and spondin 2 (SPON2). We identified 300 proteins using lectin affinity chromatography, including the glycoproteins afamin (AFM), attractin (ATRN), fibrillin 1 (FBN1), transferrin (TF), tissue inhibitor of metalloproteinase 1 (TIMP1) and vasorin (VSN). Gene ontology analysis confirmed that a majority of the identified proteins were extracellular and are mostly involved in cell communication and signaling. We also confirmed the expression of ANPEP, dickkopf WNT signaling pathway inhibitor 3 (DKK3) and osteoglycin (OGN) by multiple reaction monitoring (MRM) analysis of osteoarthritis synovial fluid samples. Conclusions We present an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis. We believe that the catalog of proteins generated in this study will further enhance our knowledge regarding the

  20. Soluble HLA in human body fluids.

    PubMed

    Aultman, D; Adamashvili, I; Yaturu, K; Langford, M; Gelder, F; Gautreaux, M; Ghali, G E; McDonald, J

    1999-03-01

    There is a growing body of information about the soluble forms of HLA in serum but there are only a few reports discussing sHLA in other body fluids. We quantitated sHLA-I and sHLA-II concentrations in sweat, saliva and tear samples from five normal individuals with known HLA-phenotypes. We also studied sweat samples from an additional 12 normal nonphenotyped subjects, as well as in CSF of 20 subjects with different illnesses, using solid phase enzyme linked immunoassay. Sweat, saliva and tears from normal subjects were found to contain very low or nondetectable amounts of sHLA-I. In contrast, sHLA-II molecules were found in each of these body fluids, although, with considerable variation between individuals. The presence of sHLA-II in saliva was further confirmed by Western-blotting. It was observed that sHLA-II having molecular mass of 43,900 and 18,100 daltons was comparable with that found in serum from normal individuals. In addition, no association of sHLA-II levels with allospecificities in either body fluid or in serum was apparent. The results of CSF sHLA concentrations in different diseases were as follows: (1) High CSF SHLA-I levels were measured during viral encephylitis (n = 3), while none of these patients contained sHLA-II in CSF; (2) The levels of sHLA-II, but not sHLA-I were elevated in CSF of patients during seizure (n = 6) and of patients with neonatal hepatitis (1 of 2) or with connective tissue disease accompanied with viral infection (n = 2); (3) No CSF sHLA-I or sHLA-II could be detected at polyneuropathy (n = 2), or in patients with syphilis (n = 3), or leukemia (n = 2) with evidence of neurologic involvement of central nervous system. Taken together, it may be concluded that the presence of sHLA in several body fluids is physiologically normal. It appears that sHLA-II is the predominant class of HLA molecules present in different body fluids. We propose that the system responsible for sHLA-II production in various body fluids must involve

  1. Increased digitalis-like activity in human cerebrospinal fluid after expansion of the extracellular fluid volume

    SciTech Connect

    Halperin, J.A.; Martin, A.M.; Malave, S.

    1985-08-12

    The present study was designed to determine whether acute expansion of the extracellular fluid volume influenced the digitalis-like activity of human cerebrospinal fluid (CSF), previously described. Human CSF samples, drawn before and 30 minutes after the intravenous infusion of 1 liter of either saline or glucose solutions, were assayed for digitalis-like activity by inhibition of either the /sup 86/Rb/sup +/ uptake into human erythrocytes or by the activity of a purified Na/sup +/-K/sup +/ ATPase. The CSF inhibitory activity on both systems significantly increased after the infusion of sodium solutions but did not change after the infusion of glucose. These results indicate that the digitalis-like factor of human CSF might be involved in the regulation of the extracellular fluid volume and electrolyte content and thereby in some of the physiological responses to sodium loading. 31 references, 2 figures, 1 table.

  2. A Cytokine-Independent Approach To Identify Antigen-Specific Human Germinal Center T Follicular Helper Cells and Rare Antigen-Specific CD4+ T Cells in Blood.

    PubMed

    Dan, Jennifer M; Lindestam Arlehamn, Cecilia S; Weiskopf, Daniela; da Silva Antunes, Ricardo; Havenar-Daughton, Colin; Reiss, Samantha M; Brigger, Matthew; Bothwell, Marcella; Sette, Alessandro; Crotty, Shane

    2016-08-01

    Detection of Ag-specific CD4(+) T cells is central to the study of many human infectious diseases, vaccines, and autoimmune diseases. However, such cells are generally rare and heterogeneous in their cytokine profiles. Identification of Ag-specific germinal center (GC) T follicular helper (Tfh) cells by cytokine production has been particularly problematic. The function of a GC Tfh cell is to selectively help adjacent GC B cells via cognate interaction; thus, GC Tfh cells may be stingy cytokine producers, fundamentally different from Th1 or Th17 cells in the quantities of cytokines produced. Conventional identification of Ag-specific cells by intracellular cytokine staining relies on the ability of the CD4(+) T cell to generate substantial amounts of cytokine. To address this problem, we have developed a cytokine-independent activation-induced marker (AIM) methodology to identify Ag-specific GC Tfh cells in human lymphoid tissue. Whereas Group A Streptococcus-specific GC Tfh cells produced minimal detectable cytokines by intracellular cytokine staining, the AIM method identified 85-fold more Ag-specific GC Tfh cells. Intriguingly, these GC Tfh cells consistently expressed programmed death ligand 1 upon activation. AIM also detected non-Tfh cells in lymphoid tissue. As such, we applied AIM for identification of rare Ag-specific CD4(+) T cells in human peripheral blood. Dengue, tuberculosis, and pertussis vaccine-specific CD4(+) T cells were readily detectable by AIM. In summary, cytokine assays missed 98% of Ag-specific human GC Tfh cells, reflecting the biology of these cells, which could instead be sensitively identified by coexpression of TCR-dependent activation markers. PMID:27342848

  3. Local fluid shifts and edema in humans during simulated microgravity

    NASA Technical Reports Server (NTRS)

    Hargens, Alan R.

    1991-01-01

    Local fluid shifts and edema in humans during simulated microgravity is studied. Recent results and significance and future plans on the following research topics are discussed: mechanisms of headward edema formation during head-down tilt; postural responses of head and foot microcirculations and their sensitivity to bed rest; and transcapillary fluid transport associated with lower body negative pressure (LBNP) with and without saline ingestion.

  4. Viral Communities Associated with Human Pericardial Fluids in Idiopathic Pericarditis

    PubMed Central

    Fancello, Laura; Monteil, Sonia; Popgeorgiev, Nikolay; Rivet, Romain; Gouriet, Frédérique; Fournier, Pierre-Edouard; Raoult, Didier; Desnues, Christelle

    2014-01-01

    Pericarditis is a common human disease defined by inflammation of the pericardium. Currently, 40% to 85% of pericarditis cases have no identified etiology. Most of these cases are thought to be caused by an infection of undetected, unsuspected or unknown viruses. In this work, we used a culture- and sequence-independent approach to investigate the viral DNA communities present in human pericardial fluids. Seven viral metagenomes were generated from the pericardial fluid of patients affected by pericarditis of unknown etiology and one metagenome was generated from the pericardial fluid of a sudden infant death case. As a positive control we generated one metagenome from the pericardial fluid of a patient affected by pericarditis caused by herpesvirus type 3. Furthermore, we used as negative controls a total of 6 pericardial fluids from 6 different individuals affected by pericarditis of non-infectious origin: 5 of them were sequenced as a unique pool and the remaining one was sequenced separately. The results showed a significant presence of torque teno viruses especially in one patient, while herpesviruses and papillomaviruses were present in the positive control. Co-infections by different genotypes of the same viral type (torque teno viruses) or different viruses (herpesviruses and papillomaviruses) were observed. Sequences related to bacteriophages infecting Staphylococcus, Enterobacteria, Streptococcus, Burkholderia and Pseudomonas were also detected in three patients. This study detected torque teno viruses and papillomaviruses, for the first time, in human pericardial fluids. PMID:24690743

  5. Aptamer-Based Screens of Human Body Fluids for Biomarkers

    PubMed Central

    Albaba, Dania; Soomro, Sanam; Mohan, Chandra

    2015-01-01

    In recent years, aptamers have come to replace antibodies in high throughput multiplexed experiments. The aptamer-based biomarker screening technology, which kicked off in 2010, is capable of interrogating thousands of proteins in a very small sample volume. With this new technology, researchers hope to find clinically appropriate biomarkers for a myriad of illnesses by screening human body fluids. In this work, we have reviewed a total of eight studies utilizing aptamer-based biomarker screens of human body fluids, and have highlighted novel protein biomarkers discovered. PMID:27600232

  6. Follicular mucinosis - Case report*

    PubMed Central

    Passos, Paola C. Vieira da Rosa; Zuchi, Manuela Ferrasso; Fabre, Andréa Buosi; Martins, Luis Eduardo A. Machado

    2014-01-01

    Follicular mucinosis, also known as alopecia mucinosa, is a cutaneous mucinosis histologically characterized by accumulation of dermal type mucin in the pilosebaceous follicle and sebaceous glands. It presents in two forms, a primary or idiopathic form and a secondary form associated with various benign or malignant processes. Among the malignant processes, the main association is with mycosis fungoides. The frequent overlap of clinical, histopathological, immunohistochemical and molecular biology characteristics makes the correct classification of these conditions difficult, therefore a long follow-up of all cases is recommended. We report the case of an adolescent with disseminated lesions and discuss the difficulty of early identification of secondary follicular mucinosis associated with cutaneous lymphoma. PMID:24770517

  7. Follicular thyroid carcinoma.

    PubMed

    Haigh, Philip I

    2002-08-01

    Follicular carcinomas are rare thyroid malignancies that are difficult to diagnose preoperatively. Fine needle aspiration is an excellent diagnostic tool and should be the initial step in managing the solitary thyroid nodule. Follicular carcinoma cannot be diagnosed with certainty by cytologic features alone; the diagnosis rests on the histologic findings of blood vessel or tumor capsule invasion. Surgical resection is the primary option for treatment. The extent of thyroidectomy for optimal survival outcome has not been determined scientifically. The outcome is excellent in minimally invasive follicular carcinoma with lobectomy and isthmusectomy; it is difficult to argue that total thyroidectomy is necessary. In a low risk prognostic group, for tumors other than minimally invasive carcinoma, lobectomy and isthmusectomy or total thyroidectomy can be justified. However, if total thyroidectomy can be done safely with a minimum of complications, then it has definite advantages for staging, postoperative surveillance, treatment, and possibly a lower recurrence rate and better survival rate. For all patients at high risk of recurrence, total thyroidectomy is preferred. PMID:12074771

  8. Antibiotic concentration in human wound fluid after intravenous administration.

    PubMed Central

    Bagley, D H; Mac Lowry, J; Beazley, R M; Gorschboth, C; Ketcham, A S

    1978-01-01

    Since the wound is the most common focus of infection in the surgical patient, adequate levels of antibiotic within the wound ar essential. This study examines the concentrations of antibiotic achieved in human wounds. Fluid was collected at timed intervals on the first postoperative day from the wounds of 56 patients receiving antibiotics after regional lymph node dissection. Antibiotic concentration was determined by bioassay. Six antibiotics were studied: cephalothin, cefazolin, cephapirin, oxacillin, ampicillin and clindamycin. The cephalosporins and penicillins showed similar patterns of appearance in the wound fluid. The peak level occurred early (1--1 1/2 hours) with subsequent slow decrease. Clindamycin produced nearly constant levels in wound fluid. The concentration of each antibiotic in wound fluid surpassed the serum levels after 2.5 hours. At the dosages studied each antibiotic produced wound fluid concentrations greater than the MIC for most susceptible organisms. Higher doses provided higher wound fluid levels. The rate of appearance and the levels achieved should be considered in the choice of antibiotics in the surgical subject. Images Fig. 1. PMID:686888

  9. Human Leukocyte Antigen Class I and II Alleles and Overall Survival in Diffuse Large B-Cell Lymphoma and Follicular Lymphoma

    PubMed Central

    Lu, Yani; Abdou, Amr M.; Cerhan, James R.; Morton, Lindsay M.; Severson, Richard K.; Davis, Scott; Cozen, Wendy; Rothman, Nathaniel; Bernstein, Leslie; Chanock, Stephen; Hartge, Patricia; Wang, Sophia S.

    2011-01-01

    Genetic variation in the 6p21 chromosomal region, including human leukocyte antigen (HLA) genes and tumor necrosis factor (TNF), has been linked to both etiology and clinical outcomes of lymphomas. We estimated the effects of HLA class I (A, B, and C), class II DRB1 alleles, and the ancestral haplotype (AH) 8.1 (HLAA*01-B*08-DRB1*03-TNF-308A) on overall survival (OS) among patients with diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) in a population-based study of non-Hodgkin lymphoma. During a median followup of 89 months, 31% (52 of 166) DLBCL and 28% (46 of 165) FL patients died. Using multivariate Cox regression models, we observed statistically significant associations between genetic variants and survival: HLA-Cw*07:01 was associated with poorer OS among DLBCL patients (Hazard ratio [HR] = 1.76, 95% confidence interval [CI] = 1.01–3.05); HLA-A*01:01 was associated with poorer OS (HR = 2.23, 95% CI = 1.24–4.01), and HLA-DRB1*13 (HR = 0.12, 95% CI = 0.02–0.90) and HLA-B Bw4 (HR = 0.36, 95% CI = 0.20–0.63) with better OS among FL patients. These results support a role for HLA in the prognosis of DLBCL and FL and represent a promising class of prognostic factors that warrants further evaluation. PMID:22125456

  10. Human leukocyte antigen class I and II alleles and overall survival in diffuse large B-cell lymphoma and follicular lymphoma.

    PubMed

    Lu, Yani; Abdou, Amr M; Cerhan, James R; Morton, Lindsay M; Severson, Richard K; Davis, Scott; Cozen, Wendy; Rothman, Nathaniel; Bernstein, Leslie; Chanock, Stephen; Hartge, Patricia; Wang, Sophia S

    2011-01-01

    Genetic variation in the 6p21 chromosomal region, including human leukocyte antigen (HLA) genes and tumor necrosis factor (TNF), has been linked to both etiology and clinical outcomes of lymphomas. We estimated the effects of HLA class I (A, B, and C), class II DRB1 alleles, and the ancestral haplotype (AH) 8.1 (HLAA*01-B*08-DRB1*03-TNF-308A) on overall survival (OS) among patients with diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) in a population-based study of non-Hodgkin lymphoma. During a median followup of 89 months, 31% (52 of 166) DLBCL and 28% (46 of 165) FL patients died. Using multivariate Cox regression models, we observed statistically significant associations between genetic variants and survival: HLA-Cw*07:01 was associated with poorer OS among DLBCL patients (Hazard ratio [HR] = 1.76, 95% confidence interval [CI] = 1.01-3.05); HLA-A*01:01 was associated with poorer OS (HR = 2.23, 95% CI = 1.24-4.01), and HLA-DRB1*13 (HR = 0.12, 95% CI = 0.02-0.90) and HLA-B Bw4 (HR = 0.36, 95% CI = 0.20-0.63) with better OS among FL patients. These results support a role for HLA in the prognosis of DLBCL and FL and represent a promising class of prognostic factors that warrants further evaluation. PMID:22125456

  11. Differential Expression of Bcl-2 Family Proteins Determines the Sensitivity of Human Follicular Lymphoma Cells to Dexamethasone-mediated and Anti-BCR-mediated Apoptosis.

    PubMed

    Adem, Jemal; Ropponen, Antti; Eeva, Jonna; Eray, Mine; Nuutinen, Ulla; Pelkonen, Jukka

    2016-01-01

    Bcl-2 family comprises proapoptotic and antiapoptotic proteins. The balance between these proteins is critical for the survival of the cells. Overexpression of the antiapoptotic protein, Bcl-2, is the hallmark of follicular lymphoma (FL). High expression of Bcl-2 provides survival advantage and may facilitate chemotherapeutic resistance in FL. In the present study, we examined expression profile of Bcl-2 family proteins such as Bcl-2, Bcl-xL, and Bim in human FL cell lines, HF1A3 and HF28. We assessed the correlation between the expression levels of these proteins and cells' sensitivity to dexamethasone (Dex)-mediated and B-cell receptor (BCR)-mediated apoptosis. Here, we show that Dex and anti-BCR-induced synergistic apoptosis which correlated with significant downregulation of Bcl-xL, inhibition of ERK1/2 phosphorylation and accumulation of nonphosphorylated Bim. However, HF28 cells were less sensitive than HF1A3 cells to Dex-induced and anti-BCR-induced apoptosis due to high Bcl-2 protein level. It is interesting to note that, a Bcl-2-specific inhibitor, ABT-199, sensitized HF28 cells to Dex-induced or anti-BCR-induced apoptosis. In addition, overexpression of Bcl-xL prevented Dex-mediated, anti-BCR-mediated, and ABT-199-mediated apoptosis, indicating that mitochondria were involved. In conclusion, these data show that the expression levels of Bcl-2 family proteins may serve to predict tumor response to BH3 mimetics and the sensitivity of FL cells to Dex-induced and anti-BCR-induced apoptosis. Moreover, our results show that BCR-targeted apoptosis might have therapeutic benefit against FL and B-cell lymphomas. PMID:26641257

  12. A critical role of T follicular helper cells in human mucosal anti-influenza response that can be enhanced by immunological adjuvant CpG-DNA.

    PubMed

    Aljurayyan, A N; Sharma, R; Upile, N; Beer, H; Vaughan, C; Xie, C; Achar, P; Ahmed, M S; McNamara, P S; Gordon, S B; Zhang, Q

    2016-08-01

    T Follicular helper cells (TFH) are considered critical for B cell antibody response, and recent efforts have focused on promoting TFH in order to enhance vaccine efficacy. We studied the frequency and function of TFH in nasopharynx-associated lymphoid tissues (NALT) from children and adults, and its role in anti-influenza antibody response following stimulation by a live-attenuated influenza vaccine (LAIV) or an inactivated seasonal virus antigen (sH1N1). We further studied whether CpG-DNA promotes TFH and by which enhances anti-influenza response. We showed NALT from children aged 1.5-10 years contained abundant TFH, suggesting efficient priming of TFH during early childhood. Stimulation by LAIV induced a marked increase in TFH that correlated with a strong production of anti-hemagglutinin (HA) IgA/IgG/IgM antibodies in tonsillar cells. Stimulation by the inactivated sH1N1 antigen induced a small increase in TFH which was markedly enhanced by CpG-DNA, accompanied by enhanced anti-HA antibody responses. In B cell co-culture experiment, anti-HA responses were only seen in the presence of TFH, and addition of plasmacytoid dendritic cell to TFH-B cell co-culture enhanced the TFH-mediated antibody production following CpG-DNA and sH1N1 antigen stimulation. Induction of TFH differentiation from naïve T cells was also shown following the stimulation. Our results support a critical role of TFH in human mucosal anti-influenza antibody response. Use of an adjuvant such as CpG-DNA that has the capacity to promote TFH by which to enhance antigen-induced antibody responses in NALT tissue may have important implications for future vaccination strategies against respiratory pathogens. PMID:27247060

  13. Development of the ovarian follicular epithelium.

    PubMed

    Rodgers, R J; Lavranos, T C; van Wezel, I L; Irving-Rodgers, H F

    1999-05-25

    A lot is known about the endocrine control of the development of ovarian follicles, but a key question now facing researchers is which molecular and cellular processes take part in control of follicular growth and development. The growth and development of ovarian follicles occurs postnatally and throughout adult life. In this review, we focus on the follicular epithelium (membrana granulosa) and its basal lamina. We discuss a model of how granulosa cells arise from a population of stem cells and then enter different lineages before differentiation. The structure of the epithelium at the antral stage of development is presented, and the effects that follicle growth has on the behavior of the granulosa cells are discussed. Finally, we discuss the evidence that during follicle development the follicular basal lamina changes in composition. This would be expected if the behavior of the granulosa cells changes, or if the permeability of the basal lamina changes. It will be evident that the follicular epithelium has similarities to other epithelia in the body, but that it is more dynamic, as gross changes occur during the course of follicle development. This basic information will be important for the development of future reproductive technologies in both humans and animals, and possibly for understanding polycystic ovarian syndrome in women. PMID:10411332

  14. Dielectric properties of human placenta, umbilical cord and amniotic fluid

    NASA Astrophysics Data System (ADS)

    Peyman, A.; Gabriel, C.; Benedickter, H. R.; Fröhlich, J.

    2011-04-01

    The dielectric properties of freshly delivered human placenta, umbilical cord and amniotic fluid have been acquired at 37 °C and in the frequency range of 200 MHz-10 GHz. The experimental data were fitted to a Cole-Cole expression. The results show that dielectric properties of the umbilical cord are significantly higher than placenta due to the presence of high water content Wharton's jelly. The results also demonstrate large differences in the dielectric properties of amniotic and cerebrospinal fluids. The data presented can be used in numerical simulations of the exposure of pregnant women to electromagnetic fields.

  15. Females with reduced fertility have excess androstenedione in follicular fluid, altered theca gene expression and increased VEGFA164b, maternal effect, and microRNA processing mRNA levels in cumulus-oocyte complexes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ovarian dysfunction contributes significantly to female infertility. However, the intrinsic and exogenous factors that result in abnormal ovarian function are poorly defined. Thus, we have established a cow model of fertility to identify mechanisms regulating follicular growth, steroidogenesis and o...

  16. Effects of weightlessness on human fluid and electrolyte physiology

    NASA Technical Reports Server (NTRS)

    Leach, Carolyn S.; Johnson, Philip C., Jr.

    1991-01-01

    Skylab and Spacelab data on changes occurring in human fluid and electrolyte physiology during the acute and adaptive phases of adaptation to spaceflight are summarized. The combined results for all three Spacelab studies show that hyponatremia developed within 20 h after the onset of weightlessness and continued throughout the flights, and hypokalemia developed by 40 h. Antidiuretic hormone was increased in plasma throughout the flights. Aldosterone decreased by 40 h, but after 7 days it had reached preflight levels.

  17. Follicular correlates with in-vitro fertilization in cattle.

    PubMed

    Greve, T; Callesen, H; Hyttel, P

    1989-01-01

    The follicular microenvironment may affect the quality and the fertilizability of an oocyte in vivo as well as in vitro. Determination of the concentrations of certain follicular fluid compounds upon aspiration could therefore be used as important clinical measures to predict the developmental ability of an oocyte submitted to in-vitro fertilization. In cattle gonadotrophin stimulation (superovulation) frequently leads to perturbed follicular steroidogenesis and oocyte maturation, and fertilization rates in vivo as well as in vitro may be reduced. These abnormal changes are monitored in the peripheral profiles of LH, progesterone and oestradiol-17 beta during the periovulatory period, and measurements of these hormones may be used to discriminate between good and poor oocyte donors. Concurrent measurements of progesterone and oestradiol-17 beta in the aspirated follicular fluids and determination of the cytogenetic features of the oocytes will reflect these deviations but, because the variation between follicles within an animal is so large, it is not possible to rely on single hormone determinations; if attempted they should be interpreted with caution. Presently, no non-invasive method such as steroid measurement and flow cytometry accurately reflects oocyte quality and function in cattle and it is seriously questioned whether such clinically applicable methods are valid. It is suggested that evaluation of oocyte quality is done on three levels: (1) the animal (peripheral hormones), (2) the ovary (stimulation, premature ovulation, follicular appearance); and (3) the follicle (cumulus oophorus complex, steroids). PMID:2677342

  18. Follicular Helper T Cells.

    PubMed

    Vinuesa, Carola G; Linterman, Michelle A; Yu, Di; MacLennan, Ian C M

    2016-05-20

    Although T cell help for B cells was described several decades ago, it was the identification of CXCR5 expression by B follicular helper T (Tfh) cells and the subsequent discovery of their dependence on BCL6 that led to the recognition of Tfh cells as an independent helper subset and accelerated the pace of discovery. More than 20 transcription factors, together with RNA-binding proteins and microRNAs, control the expression of chemotactic receptors and molecules important for the function and homeostasis of Tfh cells. Tfh cells prime B cells to initiate extrafollicular and germinal center antibody responses and are crucial for affinity maturation and maintenance of humoral memory. In addition to the roles that Tfh cells have in antimicrobial defense, in cancer, and as HIV reservoirs, regulation of these cells is critical to prevent autoimmunity. The realization that follicular T cells are heterogeneous, comprising helper and regulatory subsets, has raised questions regarding a possible division of labor in germinal center B cell selection and elimination. PMID:26907215

  19. T follicular regulatory cells.

    PubMed

    Sage, Peter T; Sharpe, Arlene H

    2016-05-01

    Pathogen exposure elicits production of high-affinity antibodies stimulated by T follicular helper (Tfh) cells in the germinal center reaction. Tfh cells provide both costimulation and stimulatory cytokines to B cells to facilitate affinity maturation, class switch recombination, and plasma cell differentiation within the germinal center. Under normal circumstances, the germinal center reaction results in antibodies that precisely target foreign pathogens while limiting autoimmunity and excessive inflammation. In order to have this degree of control, the immune system ensures Tfh-mediated B-cell help is regulated locally in the germinal center. The recently identified T follicular regulatory (Tfr) cell subset can migrate to the germinal center and inhibit Tfh-mediated B-cell activation and antibody production. Although many aspects of Tfr cell biology are still unclear, recent data have begun to delineate the specialized roles of Tfr cells in controlling the germinal center reaction. Here we discuss the current understanding of Tfr-cell differentiation and function and how this knowledge is providing new insights into the dynamic regulation of germinal centers, and suggesting more efficacious vaccine strategies and ways to treat antibody-mediated diseases. PMID:27088919

  20. Down-regulation of messenger ribonucleic acid encoding an importer of sulfoconjugated steroids during human chorionic gonadotropin-induced follicular luteinization in vivo.

    PubMed

    Brown, Kristy A; Bouchard, Nadine; Lussier, Jacques G; Sirois, Jean

    2007-01-01

    Members of the organic anion transporting polypeptide (SLCO/OATP) superfamily are capable of importing anionic compounds across the lipid bilayer in a sodium-independent manner. Member 2B1 has been shown to transport few substrates, two of which are dihydroepiandrosterone-3-sulfate (DHEA-S) and estrone-3-sulfate. Steroid sulfatase (STS) catalyses the hydrolysis of these steroids into their unconjugated counterparts. The objective of this study was to investigate the regulation of SLCO2B1 and STS mRNAs during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine SLCO2B1 cDNA was cloned and shown to encode a 709-amino acid protein (OATP2B1) that is highly conserved when compared to mammalian orthologs. RT-PCR/Southern blot analyses were performed to study the regulation of SLCO2B1 and STS transcripts in equine preovulatory follicles isolated between 0 and 39h after hCG treatment. Results showed high levels of SLCO2B1 mRNA expression before hCG, with a marked decrease observed in follicles obtained 24-39h post-hCG (P<0.05). Analyses of isolated granulosa and theca interna cells identified high mRNA expression in both cell types prior to hCG treatment, with granulosa cells showing a more rapid SLCO2B1 mRNA down-regulation. No significant change in STS mRNA was observed in intact follicle walls. However, when both cell types were isolated, a significant decrease in STS mRNA was observed in granulosa cells 24-39h post-hCG. Collectively, these results demonstrate that the hCG-dependent induction of follicular luteinization is accompanied by the down-regulation of SLCO2B1 and STS transcripts. Considering that OATP2B1 can import sulfoconjugated DHEA and estrogens, and that STS can remove the sulfonate moiety from these steroids, their down-regulation in luteinizing preovulatory follicles may provide an additional biochemical basis for the decrease in ovarian 17beta-estradiol biosynthesis after the LH surge. PMID:17049229

  1. Circulating CXCR5⁺CD4⁺ T Follicular-Like Helper Cell and Memory B Cell Responses to Human Papillomavirus Vaccines.

    PubMed

    Matsui, Ken; Adelsberger, Joseph W; Kemp, Troy J; Baseler, Michael W; Ledgerwood, Julie E; Pinto, Ligia A

    2015-01-01

    Through the interaction of T follicular helper (Tfh) cells and B cells, efficacious vaccines can generate high-affinity, pathogen-neutralizing antibodies, and memory B cells. Using CXCR5, CXCR3, CCR6, CCR7, PD1, and ICOS as markers, Tfh-like cells can be identified in the circulation and be classified into three functionally distinct subsets that are PD1+ICOS+, PD1+ ICOS-, or PD1-ICOS-. We used these markers to identify different subsets of CXCR5+CD4+ Tfh-like cells in response to highly immunogenic and efficacious vaccines for human papillomaviruses (HPV): Cervarix and Gardasil. In this small study, we used PBMC samples from 11 Gardasil recipients, and 8 Cervarix recipients from the Vaccine Research Center 902 Study to examine the induction of circulating Tfh-like cells and IgD-CD38HiCD27+ memory B cells by flow cytometry. PD1+ICOS+ CXCR3+CCR6-CXCR5+CD4+ (Tfh1-like) cells were induced and peaked on Day (D) 7 post-first vaccination, but not as much on D7 post-third vaccination. We also observed a trend toward increase in PD1+ICOS+ CXCR3-CCR6-CXCR5+CD4+ (Tfh2-like) cells for both vaccines, and PD1+ICOS+ CXCR3-CCR6+CXCR5+CD4+ (Tfh17-like) subset was induced by Cervarix post-first vaccination. There were also minimal changes in the other cellular subsets. In addition, Cervarix recipients had more memory B cells post-first vaccination than did Gardasil recipients at D14 and D30. We found frequencies of memory B cells at D30 correlated with anti-HPV16 and 18 antibody titers from D30, and the induction levels of memory B cells at D30 and PD1+ICOS+Tfh1-like cells at D7 post-first vaccination correlated for Cervarix. Our study showed that induction of circulating CXCR5+CD4+ Tfh-like subsets can be detected following immunization with HPV vaccines, and potentially be useful as a marker of immunogenicity of vaccines. However, further investigations should be extended to different cohorts with larger sample size to better understand the functions of these T cells, as well as

  2. Pulsatile cerebrospinal fluid dynamics in the human brain.

    PubMed

    Linninger, Andreas A; Tsakiris, Cristian; Zhu, David C; Xenos, Michalis; Roycewicz, Peter; Danziger, Zachary; Penn, Richard

    2005-04-01

    Disturbances of the cerebrospinal fluid (CSF) flow in the brain can lead to hydrocephalus, a condition affecting thousands of people annually in the US. Considerable controversy exists about fluid and pressure dynamics, and about how the brain responds to changes in flow patterns and compression in hydrocephalus. This paper presents a new model based on the first principles of fluid mechanics. This model of fluid-structure interactions predicts flows and pressures throughout the brain's ventricular pathways consistent with both animal intracranial pressure (ICP) measurements and human CINE phase-contrast magnetic resonance imaging data. The computations provide approximations of the tissue deformations of the brain parenchyma. The model also quantifies the pulsatile CSF motion including flow reversal in the aqueduct as well as the changes in ICPs due to brain tissue compression. It does not require the existence of large transmural pressure differences as the force for ventricular expansion. Finally, the new model gives an explanation of communicating hydrocephalus and the phenomenon of asymmetric hydrocephalus. PMID:15825857

  3. Solubility profiling of HIV protease inhibitors in human intestinal fluids.

    PubMed

    Wuyts, Benjamin; Brouwers, Joachim; Mols, Raf; Tack, Jan; Annaert, Pieter; Augustijns, Patrick

    2013-10-01

    The present study pursued to profile the intestinal solubility of nine HIV protease inhibitors (PIs) in fasted- and fed-state human intestinal fluids (FaHIF, FeHIF) aspirated from four volunteers. In addition, the ability of fasted- and fed-state simulated intestinal fluids (FaSSIF, FeSSIF) to predict the intestinal solubility was evaluated. All PIs were poorly soluble in FaHIF (from 7 μM for ritonavir to 327 μM for darunavir) and FeHIF (from 15 μM for atazanavir to 409μM for darunavir). For four of nine PIs, food intake significantly enhanced the solubilizing capacity of intestinal fluids (up to 18.4-fold increase for ritonavir). The intersubject variability (average coefficient of variance CVfed = 60.6%, CVfasted = 40.4%) was higher as compared with the intrasubject variability (CVfed = 41.3%, CVfasted = 20.5%). PI solubilities correlated reasonably well between FaSSIF and FaHIF (R = 0.817), but not between FeSSIF and FeHIF (R = 0.617). To conclude, postprandial conditions increased the inter- and intrasubject variability of the PIs. The inability of FeSSIF to accurately predict the FeHIF solubility emphasizes the need for a multivariate approach to determine solubility profiles, taking into account solid-state characteristics, pH, mixed bile acid/phospholipid micelles, and digestive products. PMID:23939880

  4. Follicular dynamics in Mangalarga mares.

    PubMed

    Buratini, J; Rosa e Silva, A A; Barros, C M; Papa, F O; Caldas, M C; Meira, C

    1997-12-01

    Ovarian follicular activity was studied by ultrasonography during 17 oestrous cycles in 9 Mangalarga mares during the second half of the ovulatory season. Sixteen oestrous cycles were considered normal and one 3-wave cycle showing a prolonged luteal phase was considered atypical. Daily ultrasonographic examinations were performed and the compiled data on follicular dynamics were studied retrospectively. One major wave of follicular growth was observed in 13 of the 16 normal cycles (81.25%), whereas 2 major waves occurred in 3 cycles (18.75%). The mean (+/- s.d.) days of emergence of the primary wave of follicular development in cycles containing one or 2 waves were Day 6.0 +/- 2.3 and Day 11.0 +/- 1.0, respectively. The secondary wave of follicular development in 2-wave cycles emerged on Day 0.0 +/- 3.6. The day of wave divergence for primary waves of follicular development in cycles which exhibited one or 2 major waves were Day 12.2 +/- 3.5 and Day 17.3 +/- 3.0, respectively. Divergence of secondary waves occurred in only one of the 3 cycles which exhibited 2 major follicular waves (Day 7). The mean (+/- s.d.) maximum diameters of the dominant follicle in the primary wave of oestrous cycles exhibiting one and 2 major waves were 39.0 +/- 3.9 mm and 34.7 +/- 2.5 mm, respectively. The mean (+/- s.d.) maximum diameter of the dominant follicle present in the secondary wave was 34.3 +/- 11.0 mm. The mean (+/- s.d.) lengths of the interovulatory intervals for cycles containing one and 2 major waves were 19.4 +/- 2.2 and 23.3 +/- 2.5 days, respectively. These data indicate that most Mangalarga mares show one major follicular wave during the oestrous cycle but a small percentage of mares show 2 major waves. PMID:9593519

  5. Follicular Dendritic Cell Sarcoma

    PubMed Central

    Udayakumar, Achandira M.; Al-Bahri, Maiya; Burney, Ikram A.; Al-Haddabi, Ibrahim

    2015-01-01

    Follicular dendritic cell sarcoma (FDCS) is a rare neoplasm with a non-specific and insidious presentation further complicated by the difficult diagnostic and therapeutic assessment. It has a low to intermediate risk of recurrence and metastasis. Unlike other soft tissue sarcomas or histiocytic and dendritic cell neoplasms, cytogenetic studies are very limited in FDCS cases. Although no specific chromosomal marker has yet been established, complex aberrations and different ploidy types have been documented. We report the case of a 39-year-old woman with FDCS who presented to the Sultan Qaboos University Hospital in Muscat, Oman, in February 2013. Ultrastructural, immunophenotypical and histological findings are reported. In addition, karyotypic findings showed deletions of the chromosomes 1p, 3q, 6q, 7q, 8q and 11q. To the best of the authors’ knowledge, these have not been reported previously in this tumour. Techniques such as spectral karyotyping may help to better characterise chromosomal abnormalities in this type of tumour. PMID:26355964

  6. PROTEOMIC ANALYSIS OF HUMAN BRONCHOALVEOLAR LAVAGE FLUID AFTER SUBSGEMENTAL EXPOSURE

    PubMed Central

    Foster, Matthew W.; Will Thompson, J.; Que, Loretta G.; Yang, Ivana V.; Schwartz, David A.; Arthur Moseley, M.; Marshall, Harvey E.

    2013-01-01

    The analysis of airway fluid, as sampled by bronchoalveolar lavage (BAL), provides a minimally invasive route to interrogate lung biology in health and disease. Here, we used immunodepletion, coupled with gel- and label-free LC-MS/MS, for quantitation of the BAL fluid (BALF) proteome in samples recovered from human subjects following bronchoscopic instillation of saline, lipopolysaccharide (LPS) or house dust mite antigen into three distinct lung subsegments. Among more than 200 unique proteins quantified across nine samples, neutrophil granule-derived and acute phase proteins were most highly enriched in the LPS-exposed lobes. Of these, peptidoglycan response protein 1 was validated and confirmed as a novel marker of neutrophilic inflammation. Compared to a prior transcriptomic analysis of airway cells in this same cohort, the BALF proteome revealed a novel set of response factors. Independent of exposure, the enrichment of tracheal-expressed proteins in right lower lung lobes suggests a potential for constitutive intralobar variability in the BALF proteome; sampling of multiple lung subsegments also appears to aid in the identification of protein signatures that differentiate individuals at baseline. Collectively, this proof-of-concept study validates a robust workflow for BALF proteomics and demonstrates the complementary nature of proteomic and genomic techniques for investigating airway (patho)physiology. PMID:23550723

  7. Effects of Weightlessness on Human Fluid and Electrolyte Physiology

    NASA Technical Reports Server (NTRS)

    Leach, Carolyn S.; Johnson, Philip C., Jr.

    1991-01-01

    The changes that occur in human fluid and electrolyte physiology during the acute and adaptive phases of adaptation to spaceflight are summarized. A number of questions remain to be answered. At a time when plasma volume and extracellular fluid volume are contracted and salt and water intake is unrestricted. ADH does not correct the volume deficit and serum sodium decreases. Change in secretion or activity of a natriuretic factor during spaceflight is one possible explanation. Recent identification of a polypeptide hormone produced in cardiac muscle cells which is natiuretic, is hypotensive, and has an inhibitory effect on renin and aldosterone secretion has renewed interest in the role of a natriuretic factor. The role of this atrial natriuretic factor (ANF) in both long- and short-term variation in extracellular volumes and in the inability of the kidney to bring about an escape from the sodium-retaining state accompanying chronic cardiac dysfunction makes it reasonable to look for a role of ANF in the regulation of sodium during exposure to microgravity. Prostaglandin-E is another hormone that may antagonize the action of ADH. Assays of these hormones will be performed on samples from crew members in the future.

  8. Improved analysis of malondialdehyde in human body fluids.

    PubMed

    Jentzsch, A M; Bachmann, H; Fürst, P; Biesalski, H K

    1996-01-01

    The widely used TBA assay for lipid peroxidation was modified to minimize artefactual oxidative degradation of lipids during the assay. Formation of the TBA-MDA condensation product was studied with and without exclusion of oxygen, and the concentration effect of BHT addition was examined. Oxygen was depleted from the reaction mixture by extensive argon gassing. Exclusion of oxygen resulted in decreased TBARS production in plasma but not in standard solutions. High BHT concentrations resulted in a similar effect. At concentrations higher than 3 mmol/l BHT exclusion of oxygen had no additional effect. By measuring n-butanol extracts in a multititer plate reader this modified method was made suitable as a preliminary screening assay of human body fluids for lipid peroxidation. PMID:8746446

  9. Reactive oxygen species (ROS): involvement in bovine follicular cysts etiopathogenesis.

    PubMed

    Rizzo, Annalisa; Minoia, Giuseppe; Trisolini, Carmelinda; Mutinati, Maddalena; Spedicato, Massimo; Jirillo, Felicita; Sciorsci, Raffaele Luigi

    2009-01-01

    Ovulation is compared to an acute inflammatory process during which vasoactive agents, prostanoids, leukotrienes and Reactive Oxygen Species (ROS) develop. The aim of this study was to evaluate the levels of ROS in cystic and follicular fluid, in order to establish their involvement in the etiopathogenesis of Cystic Ovarian Follicle (COF) in dairy cows. The study was conducted in 30 healthy cows (group C) and 30 cows affected by COF (group COF). The fluid of follicular cysts and of preovulatory follicles was drawn by means of ultrasound guided aspiration from the cows of both groups. The fluid obtained was analyzed by a photometric analytical system to detect ROS level. ROS concentration was statistically lower in the cystic fluid than in the follicular one (62.4 +/- 13.36 U.Carr vs. 84.89 +/- 26.99 U.Carr) (p<0.05), thus suggesting that an alteration of the cascade responsible for ROS production may be implicated in the complex etipathogenesis of COF. PMID:19874233

  10. Intestinal drug solubility estimation based on simulated intestinal fluids: comparison with solubility in human intestinal fluids.

    PubMed

    Clarysse, Sarah; Brouwers, Joachim; Tack, Jan; Annaert, Pieter; Augustijns, Patrick

    2011-07-17

    The purpose of this study was to validate both existing fasted and fed state simulated intestinal fluids (FaSSIF and FeSSIF), and simpler, alternative media for predicting intraluminal drug solubility during drug discovery and early drug development. For 17 model drugs, the solubilizing capacity of FaSSIF(c) and FeSSIF(c) (subscript indicates the use of crude taurocholate) and different concentrations of D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) in phosphate buffer were correlated with the solubilizing capacity of human intestinal fluids (HIF) in the fasted and the early postprandial state. A good correlation between solubility in fasted HIF and FaSSIF(c) and between solubility in fed HIF and FeSSIF(c) was obtained, indicated by R(2) values of 0.91 and 0.86, respectively. Comparable values were obtained for 0.1% TPGS for the fasted state (R(2)=0.84) and 2% TPGS for the fed state (R(2)=0.84). Direct estimation of intestinal drug solubility by the measured solubilities in FaSSIF(c) and FeSSIF(c) was acceptable. However, better estimates were obtained by calculating solubilities based on the equations describing the relationship between solubilities in FaSSIF(c) and FeSSIF(c) as function of observed solubilities in HIF. Using this approach, the predictive value of the TPGS-based solvent system was also acceptable and comparable to that of FaSSIF(c) and FeSSIF(c). In conclusion, FaSSIF(c) and FeSSIF(c) can be considered biorelevant media for intestinal solubility estimation. A simpler TPGS-based system may be a valuable alternative with improved stability and lower cost. PMID:21570465

  11. Adaptation of Group A Streptococcus to Human Amniotic Fluid

    PubMed Central

    Sitkiewicz, Izabela; Green, Nicole M.; Guo, Nina; Bongiovanni, Ann M.; Witkin, Steven S.; Musser, James M.

    2010-01-01

    Background For more than 100 years, group A Streptococcus has been identified as a cause of severe and, in many cases, fatal infections of the female urogenital tract. Due to advances in hospital hygiene and the advent of antibiotics, this type of infection has been virtually eradicated. However, within the last three decades there has been an increase in severe intra- and post-partum infections attributed to GAS. Methodology We hypothesized that GAS alters its transcriptome to survive in human amniotic fluid (AF) and cause disease. To identify genes that were up or down regulated in response to growth in AF, GAS was grown in human AF or standard laboratory media (THY) and samples for expression microarray analysis were collected during mid-logarithmic, late-logarithmic, and stationary growth phases. Microarray analysis was performed using a custom Affymetrix chip and normalized hybridization values derived from three biological replicates were collected at each growth point. Ratios of AF/THY above a 2-fold change and P-value <0.05 were considered significant. Principal Findings The majority of changes in the GAS transcriptome involved down regulation of multiple adhesins and virulence factors and activation of the stress response. We observed significant changes in genes involved in the arginine deiminase pathway and in the nucleotide de novo synthesis pathway. Conclusions/Significance Our work provides new insight into how pathogenic bacteria respond to their environment to establish infection and cause disease. PMID:20352104

  12. Follicular Unit Extraction Hair Transplant

    PubMed Central

    Dua, Aman; Dua, Kapil

    2010-01-01

    Hair transplantation has come a long way from the days of Punch Hair Transplant by Dr. Orentreich in 1950s to Follicular Unit Hair Transplant (FUT) of 1990s and the very recent Follicular Unit Extraction (FUE) technique. With the advent of FUE, the dream of ‘no visible scarring’ in the donor area is now looking like a possibility. In FUE, the grafts are extracted as individual follicular units in a two-step or three-step technique whereas the method of implantation remains the same as in the traditional FUT. The addition of latest automated FUE technique seeks to overcome some of the limitations in this relatively new technique and it is now possible to achieve more than a thousand grafts in one day in trained hands. This article reviews the methodology, limitations and advantages of FUE hair transplant. PMID:21031064

  13. The role of amniotic fluid in force transfer during human birth

    NASA Astrophysics Data System (ADS)

    Baumer, Alexa; Lehn, Andrea; Leftwich, Megan

    2013-11-01

    This study seeks to understand the fundamental fluid dynamic processes involved in human birth. We begin by examining the importance of amniotic fluid. This is done using two experimental techniques that approximate the laboring human uterus to different degrees of anatomical correctness. The first, in which a latex uterus is filled with fluid and a solid fetus is extracted, investigates the importance of both amniotic fluid properties and fetal position in the force required to remove a fetus. The second experiment simplifies the geometry of birth even more. In this case, a solid cylindrical rod is pulled through a highly flexible outer tube. The force to pull the inner cylinder as a function of the gap fluid properties is measured. By carefully controlling the fluid properties of the experiment, the study will provide further insight into the roles of amniotic fluid in human birth.

  14. Inverted follicular keratosis successfully treated with imiquimod.

    PubMed

    Karadag, Ayse Serap; Ozlu, Emin; Uzuncakmak, Tugba Kevser; Akdeniz, Necmettin; Cobanoglu, Bengu; Oman, Berkant

    2016-01-01

    Inverted follicular keratosis is a rare benign tumor of the follicular infundibulum characterized by exo-endophytic growing. It is thought to be a rare variant of the seborrheic keratosis. The diagnosis of inverted follicular keratosis is generally established histopathologically because clinical differentiation from other lesions is difficult. Herein, we present one such rare case, successfully treated with topical 5% imiquimod cream. PMID:27294052

  15. Inverted follicular keratosis successfully treated with imiquimod

    PubMed Central

    Karadag, Ayse Serap; Ozlu, Emin; Uzuncakmak, Tugba Kevser; Akdeniz, Necmettin; Cobanoglu, Bengu; Oman, Berkant

    2016-01-01

    Inverted follicular keratosis is a rare benign tumor of the follicular infundibulum characterized by exo-endophytic growing. It is thought to be a rare variant of the seborrheic keratosis. The diagnosis of inverted follicular keratosis is generally established histopathologically because clinical differentiation from other lesions is difficult. Herein, we present one such rare case, successfully treated with topical 5% imiquimod cream. PMID:27294052

  16. Human cervicovaginal fluid biomarkers to predict term and preterm labor

    PubMed Central

    Heng, Yujing J.; Liong, Stella; Permezel, Michael; Rice, Gregory E.; Di Quinzio, Megan K. W.; Georgiou, Harry M.

    2015-01-01

    Preterm birth (PTB; birth before 37 completed weeks of gestation) remains the major cause of neonatal morbidity and mortality. The current generation of biomarkers predictive of PTB have limited utility. In pregnancy, the human cervicovaginal fluid (CVF) proteome is a reflection of the local biochemical milieu and is influenced by the physical changes occurring in the vagina, cervix and adjacent overlying fetal membranes. Term and preterm labor (PTL) share common pathways of cervical ripening, myometrial activation and fetal membranes rupture leading to birth. We therefore hypothesize that CVF biomarkers predictive of labor may be similar in both the term and preterm labor setting. In this review, we summarize some of the existing published literature as well as our team's breadth of work utilizing the CVF for the discovery and validation of putative CVF biomarkers predictive of human labor. Our team established an efficient method for collecting serial CVF samples for optimal 2-dimensional gel electrophoresis resolution and analysis. We first embarked on CVF biomarker discovery for the prediction of spontaneous onset of term labor using 2D-electrophoresis and solution array multiple analyte profiling. 2D-electrophoretic analyses were subsequently performed on CVF samples associated with PTB. Several proteins have been successfully validated and demonstrate that these biomarkers are associated with term and PTL and may be predictive of both term and PTL. In addition, the measurement of these putative biomarkers was found to be robust to the influences of vaginal microflora and/or semen. The future development of a multiple biomarker bed-side test would help improve the prediction of PTB and the clinical management of patients. PMID:26029118

  17. Excipient-mediated supersaturation stabilization in human intestinal fluids.

    PubMed

    Bevernage, Jan; Forier, Thomas; Brouwers, Joachim; Tack, Jan; Annaert, Pieter; Augustijns, Patrick

    2011-04-01

    It was the purpose of this study to investigate excipient-mediated precipitation inhibition upon induction of supersaturation of poorly water-soluble drugs in aspirated human intestinal fluids (HIF) representing both the fasted and fed state. Etravirine, ritonavir, loviride, danazol and fenofibrate were selected as model compounds. For comparative purposes, precipitation inhibition was also evaluated in simple aqueous buffer, and in intestinal simulation media representative for the fasted and fed state (FaSSIF and FeSSIF, respectively). Supersaturation was induced in the test media containing predissolved excipient (HPMC-AS, HPMC-E5, HPMC-E50, HPMC-E4M, HPMC-P and PVP) at a defined degree of supersaturation (DS = 20) using the solvent shift method. The results illustrate that cellulosic polymers can reduce the precipitation rate and stabilize supersaturation in HIF. The extent of stabilization was compound and excipient dependent but independent of the nutritional state. Whenever excipient effects were observed, the predictive value of simple buffer or FaSSIF/FeSSIF was rather limited. In general, excipient-mediated precipitation inhibition was less pronounced in HIF compared to simple aqueous buffer or FaSSIF/FeSSIF. However, excipients showing no effect in simple aqueous buffer or FaSSIF/FeSSIF also proved to be ineffective in HIF, indicating the value of these simulation media in the elimination of excipients during formulation development. PMID:21268663

  18. Exploration of the normal human bronchoalveolar lavage fluid proteome

    PubMed Central

    Chen, Jinzhi; Ryu, Soyoung; Gharib, Sina A.; Goodlett, David R.; Schnapp, Lynn M.

    2015-01-01

    We obtained insight into normal lung function by proteome analysis of bronchoalveolar lavage fluid (BALF) from six normal human subjects using a “Lyse-N-Go’ shotgun proteomic protocol. Intra-sample variation was calculated using three different label-free methods, (i) protein sequence coverage; (ii) peptide spectral counts and (iii) peptide single-ion current areas (PICA), which generates protein expression data by summation of the area under the curve for a given peptide single-ion current trace and then adding values for all peptides from that same parent protein. PICA gave the least intra-subject variability and was used to calculate differences in protein expression between the six subjects. We observed an average threefold inter-sample variability, which affects analysis of changes in protein expression that occur in different diseases. We detected 167 unique proteins with >100 proteins detected in each of the six individual BAL samples, 42 of which were common to all six subjects. Gene ontology analysis demonstrated enrichment of several biological processes in the lung, reflecting its expected role in gas exchange and host defense as an immune organ. The same biological processes were enriched compared to either plasma or total genome proteome, suggesting an active enrichment of plasma proteins in the lung rather than passive capillary leak. PMID:21136857

  19. Sphingolipids in Human Synovial Fluid - A Lipidomic Study

    PubMed Central

    Kosinska, Marta Krystyna; Liebisch, Gerhard; Lochnit, Guenter; Wilhelm, Jochen; Klein, Heiko; Kaesser, Ulrich; Lasczkowski, Gabriele; Rickert, Markus; Schmitz, Gerd; Steinmeyer, Juergen

    2014-01-01

    Articular synovial fluid (SF) is a complex mixture of components that regulate nutrition, communication, shock absorption, and lubrication. Alterations in its composition can be pathogenic. This lipidomic investigation aims to quantify the composition of sphingolipids (sphingomyelins, ceramides, and hexosyl- and dihexosylceramides) and minor glycerophospholipid species, including (lyso)phosphatidic acid, (lyso)phosphatidylglycerol, and bis(monoacylglycero)phosphate species, in the SF of knee joints from unaffected controls and from patients with early (eOA) and late (lOA) stages of osteoarthritis (OA), and rheumatoid arthritis (RA). SF without cells and cellular debris from 9 postmortem donors (control), 18 RA, 17 eOA, and 13 lOA patients were extracted to measure lipid species using electrospray ionization tandem mass spectrometry - directly or coupled with hydrophilic interaction liquid chromatography. We provide a novel, detailed overview of sphingolipid and minor glycerophospholipid species in human SF. A total of 41, 48, and 50 lipid species were significantly increased in eOA, lOA, and RA SF, respectively when compared with normal SF. The level of 21 lipid species differed in eOA SF versus SF from lOA, an observation that can be used to develop biomarkers. Sphingolipids can alter synovial inflammation and the repair responses of damaged joints. Thus, our lipidomic study provides the foundation for studying the biosynthesis and function of lipid species in health and most prevalent joint diseases. PMID:24646942

  20. Risk assessment of thyroid follicular cell tumors.

    PubMed Central

    Hill, R N; Crisp, T M; Hurley, P M; Rosenthal, S L; Singh, D V

    1998-01-01

    Thyroid follicular cell tumors arise in rodents from mutations, perturbations of thyroid and pituitary hormone status with increased stimulation of thyroid cell growth by thyroid-stimulating hormone (TSH), or a combination of the two. The only known human thyroid carcinogen is ionizing radiation. It is not known for certain whether chemicals that affect thyroid cell growth lead to human thyroid cancer. The U.S. Environmental Protection Agency applies the following science policy positions: 1) chemically induced rodent thyroid tumors are presumed to be relevant to humans; 2) when interspecies information is lacking, the default is to assume comparable carcinogenic sensitivity in rodents and humans; 3) adverse rodent noncancer thyroid effects due to chemically induced thyroid-pituitary disruption are presumed to be relevant to humans; 4) linear dose-response considerations are applied to thyroid cancer induced by chemical substances that either do not disrupt thyroid functioning or lack mode of action information; 5) nonlinear thyroid cancer dose-response considerations are applied to chemicals that reduce thyroid hormone levels, increase TSH and thyroid cell division, and are judged to lack mutagenic activity; and 6) nonlinear considerations may be applied in thyroid cancer dose-response assessments on a case-by-case basis for chemicals that disrupt thyroid-pituitary functioning and demonstrate some mutagenic activity. Required data for risk assessment purposes is mode of action information on mutagenicity, increases in follicular cell growth (cell size and number) and thyroid gland weight, thyroid-pituitary hormones, site of action, correlations between doses producing thyroid effects and cancer, and reversibility of effects when dosing ceases. Images Figure 1 Figure 2 Figure 3 PMID:9681971

  1. Early Rise of Blood T Follicular Helper Cell Subsets and Baseline Immunity as Predictors of Persisting Late Functional Antibody Responses to Vaccination in Humans

    PubMed Central

    Borgogni, Erica; Zedda, Luisanna; Cantisani, Rocco; Chiappini, Nico; Schiavetti, Francesca; Rosa, Domenico; Castellino, Flora; Montomoli, Emanuele; Bodinham, Caroline L.; Lewis, David J.; Medini, Duccio; Bertholet, Sylvie; Del Giudice, Giuseppe

    2016-01-01

    CD4+ T follicular helper cells (TFH) have been identified as the T-cell subset specialized in providing help to B cells for optimal activation and production of high affinity antibody. We recently demonstrated that the expansion of peripheral blood influenza-specific CD4+IL-21+ICOS1+ T helper (TH) cells, three weeks after vaccination, associated with and predicted the rise of protective neutralizing antibodies to avian H5N1. In this study, healthy adults were vaccinated with plain seasonal trivalent inactivated influenza vaccine (TIIV), MF59®-adjuvanted TIIV (ATIIV), or saline placebo. Frequencies of circulating CD4+ TFH1 ICOS+ TFH cells and H1N1-specific CD4+IL-21+ICOS+ CXCR5+ TFH and CXCR5- TH cell subsets were determined at various time points after vaccination and were then correlated with hemagglutination inhibition (HI) titers. All three CD4+ T cell subsets expanded in response to TIIV and ATIIV, and peaked 7 days after vaccination. To demonstrate that these TFH cell subsets correlated with functional antibody titers, we defined an alternative endpoint metric, decorrelated HI (DHI), which removed any correlation between day 28/day 168 and day 0 HI titers, to control for the effect of preexisting immunity to influenza vaccine strains. The numbers of total circulating CD4+ TFH1 ICOS+ cells and of H1N1-specific CD4+IL-21+ICOS+ CXCR5+, measured at day 7, were significantly associated with day 28, and day 28 and 168 DHI titers, respectively. Altogether, our results show that CD4+ TFH subsets may represent valuable biomarkers of vaccine-induced long-term functional immunity. Trial Registration ClinicalTrials.gov NCT01771367 PMID:27336786

  2. Crowding and Follicular Fate: Spatial Determinants of Follicular Reserve and Activation of Follicular Growth in the Mammalian Ovary

    PubMed Central

    Gaytan, Francisco; Morales, Concepcion; Leon, Silvia; Garcia-Galiano, David; Roa, Juan; Tena-Sempere, Manuel

    2015-01-01

    Initiation of growth of resting ovarian follicles is a key phenomenon for providing an adequate number of mature oocytes in each ovulation, while preventing premature exhaustion of primordial follicle reserve during the reproductive lifespan. Resting follicle dynamics strongly suggest that primordial follicles are under constant inhibitory influences, by mechanisms and factors whose nature remains ill defined. In this work, we aimed to assess the influence of spatial determinants, with special attention to clustering patterns and crowding, on the fate of early follicles in the adult mouse and human ovary. To this end, detailed histological and morphometric analyses, targeting resting and early growing follicles, were conducted in ovaries from mice, either wild type (WT) or genetically modified to lack kisspeptin receptor expression (Kiss1r KO), and healthy adult women. Kiss1r KO mice were studied as model of persistent hypogonadotropism and anovulation. Different qualitative and quantitative indices of the patterns of spatial distribution of resting and early growing follicles in the mouse and human ovary, including the Morisita’s index of clustering, were obtained. Our results show that resting primordial follicles display a clear-cut clustered pattern of spatial distribution in adult mouse and human ovaries, and that resting follicle aggrupation is inversely correlated with the proportion of follicles initiating growth and entering into the growing pool. As a whole, our data suggest that resting follicle crowding, defined by changes in density and clustered pattern of distribution, is a major determinant of follicular activation and the fate of ovarian reserve. Uneven follicle crowding would constitute the structural counterpart of the major humoral regulators of early follicular growth, with potential implications in ovarian ageing and pathophysiology. PMID:26642206

  3. Cytopathology of Follicular Cell Nodules.

    PubMed

    Damiani, Domenico; Suciu, Voichita; Vielh, Philippe

    2015-12-01

    This article corresponds to a lecture delivered during the Endocrine Pathology Society symposium held in Boston on 21 March 2015 (104th USCAP meeting, March 21-27). It focuses on the importance of cytopathology in endocrine thyroid pathology and the limits and pitfalls of diagnosis in follicular cell lesions. Lights and shadows are present in each diagnostic technique: Fine needle aspiration has imposed itself as a gold standard in thyroid nodules thanks to its easiness of execution and high cost-effectiveness ratio. A milestone in this field is represented by the National Cancer Institute (NCI) Thyroid Fine Needle Aspiration (FNA) State of the State of the Science Conference hosted in October 22-23, 2007 by the NCI, followed by a series of documents published in Diagnostic Cytopathology and Cytojournal (2008) as well as in an atlas entitled: The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC): terminology and criteria (2010, Springer). "Gray" zones still remain, causing difficulties and anxiety to the cytopathologist when facing challenging cases. Each diagnostic category of TBSRTC is analyzed and discussed in a concise fashion with special emphasis on challenging cases such as atypia of undetermined significance (AUS), suspicion for follicular neoplasms (SFNs), diagnoses of papillary thyroid carcinoma (PTC) in Hashimoto thyroiditis and follicular variant of papillary carcinoma (FVPTC). Our aim was to better define and clarify the spectrum of follicular cell lesions in thyroid nodule samplings and to underline the diagnostic limits in order to avoid pitfalls. New emerging molecular biology techniques may represent useful tools in selected morphological challenging cases and lead to new therapeutic approaches in line with drug-tailored therapy and personalized medicine. PMID:26227345

  4. Evaluation of Human Body Fluids for the Diagnosis of Fungal Infections

    PubMed Central

    2013-01-01

    Invasive fungal infections are a major cause of morbidity and mortality in immunocompromised patients. Because the etiologic agents of these infections are abundant in nature, their isolation from biopsy material or sterile body fluids is needed to document infection. This review evaluates and discusses different human body fluids used to diagnose fungal infections. PMID:23984401

  5. Bilateral conjunctival follicular lymphoma in a child.

    PubMed

    Wall, Palak B; Traboulsi, Elias I; Hsi, Eric D; Singh, Arun D

    2015-04-01

    Follicular lymphoma is exceedingly rare in children. We present the case of a 10-year-old patient with a conjunctival lesion on the left eye who later developed a similar lesion on the right eye. Excisional biopsy of the left eye lesion revealed follicular lymphoma. The patient was treated with systemic rituximab. To our knowledge, only 4 other cases of pediatric conjunctival follicular lymphoma have been reported, all of which were isolated lesions that were treated with excisional biopsy alone. PMID:25824110

  6. ADDRESSING HUMAN EXPOSURES TO AIR POLLUTANTS AROUND BUILDINGS IN URBAN AREAS WITH COMPUTATIONAL FLUID DYNAMICS MODELS

    EPA Science Inventory

    This paper discusses the status and application of Computational Fluid Dynamics (CFD) models to address challenges for modeling human exposures to air pollutants around urban building microenvironments. There are challenges for more detailed understanding of air pollutant sour...

  7. Is AMH a regulator of follicular atresia?

    PubMed

    Seifer, David B; Merhi, Zaher

    2014-11-01

    We discuss the hypothesis that AMH is an intraovarian regulator that inhibits follicular atresia within the human ovary. Several indirect lines of evidence derived from clinical and basic science studies in a variety of different patient populations and model systems collectively support this hypothesis. Evidence presented herein include 1) timing of onset of menopause in women with polycystic ovary syndrome, 2) site of cellular origin and timing of AMH production, 3) AMH's influence on other critical growth factors and enzymes involved in folliculogenesis, and 4) AMH's inhibition of granulosa apoptosis. If this hypothesis is true, it may provide insight for treatment strategies for prevention and treatment of premature ovarian insufficiency, slowing natural ovarian aging, and/or delaying eventual ovarian failure. Such findings may lead to the development of 1) AMH agonists for retarding the onset of menopause and/or as a chemoprotectant prior to cancer therapy and 2) AMH antagonists for the treatment of PCOS. PMID:25193290

  8. Regulation of the ovarian follicular vasculature

    PubMed Central

    Fraser, Hamish M

    2006-01-01

    Angiogenesis is associated with follicular development and is regulated independently within each follicle potentially making the functioning of its vasculature critically important in determining its fate. This review examines the various ways in which follicular angiogenesis may be monitored, describes the follicular localisation and changes in pro- and anti-angiogenic factors that may regulate the process and how antagonists may be used to elucidate their physiological role in vivo. Thus, inhibition of vascular endothelial growth factor (VEGF), VEGF receptor-2, vascular endothelial cell cadherin or interference with the angiopoietin system can inhibit follicular development or prevent ovulation. PMID:16611363

  9. Size heterogeneity of epidermal growth factor in human body fluids

    SciTech Connect

    Pesonen, K.; Viinikka, L.; Koskimies, A.; Banks, A.R.; Nicolson, M.; Perheentupa, J.

    1987-06-29

    The authors measured the concentration of immunoreactive (IR) hEGF in various body fluids by radioimmunoassay (RIA) and evaluated its size heterogeneity by size exclusion high performance liquid chromatography combined with RIA or with time-resolved immunofluorometric assay (TR-IFMA). Mean concentration was 80 ng/ml in urine, 65 ng/ml in milk, 50 ng/ml in seminal plasma, 25 ng/ml in armpit sweat, 1 ng/ml in breast sweat, 0.3 ng/ml in third-trimester amniotic fluid, 3 ng/ml in saliva, 1.5 ng/ml in tears and 0.3 ng/ml in gastric juice. All the fluids except armpit sweat and gastric juice contained two to five molecular sizes of IR-hEGF. As well as the 6200-dalton (6.2kDa) hEGF the authors found at least four other different molecular sizes with approximate weights of greater than or equal to300, 150, 70 and 20 kDa. The authentic 6.2kDa form made up >90% of the total IR-hEGF in all except the amniotic fluid where its proportion was 71%, and the seminal plasma where the proportion could not be determined. 18 references, 1 figure, 1 table.

  10. Inactivation of human immunodeficiency virus (HIV) by ionizing radiation in body fluids and serological evidence

    SciTech Connect

    Bigbee, P.D.; Sarin, P.S.; Humphreys, J.C.; Eubanks, W.G.; Sun, D.; Hocken, D.G.; Thornton, A.; Adams, D.E.; Simic, M.G. )

    1989-11-01

    A method to use ionizing radiation to inactivate HIV (Human Immunodeficiency Virus) in human body fluids was studied in an effort to reduce the risk of accidental infection to forensic science laboratory workers. Experiments conducted indicate that an X-ray absorbed dose of 25 krad was required to completely inactivate HIV. This does not alter forensically important constituents such as enzymes and proteins in body fluids. This method of inactivation of HIV cannot be used on body fluids which will be subjected to deoxyribonucleic acid (DNA) typing.

  11. Complement-derived leukotactic factors in inflammatory synovial fluids of humans

    PubMed Central

    Ward, Peter A.; Zvaifler, Nathan J.

    1971-01-01

    A large per cent of rheumatoid synovial fluids contain chemotactic activity for rabbit granulocytes (neutrophilic). The chemotactic activity is, in large part, related to the fifth (C5) and sixth (C6) components of human complement; a combination of physical-chemical techniques indicates the activity to be attributable to C567 and C5a, a cleavage product of C5. Many rheumatoid synovial fluids contain a C5-cleaving enzyme which, on the basis of substrate specificity and susceptibility to inhibitors, is very similar to an enzyme extractable from lysosomal granules of human and rabbit granulocytes. Inflammatory nonrheumatoid synovial fluids contain chemotactic activity that is related to cleavage products (C3a) of the third component of human complement (C3). Also found in these fluids is a C3-cleaving enzyme capable of producing C3a. Of the other synovial fluids examined, lupus fluids were remarkable by their total lack of chemotactic activity. These findings record for the first time the presence of complement-derived chemotactic factors in pathological human fluids. Images PMID:5545123

  12. Absorbent product to absorb fluids. [for collection of human wastes

    NASA Technical Reports Server (NTRS)

    Dawn, F. S.; Correale, J. V. (Inventor)

    1982-01-01

    A multi-layer absorbent product for use in contact with the skin to absorb fluids is discussed. The product utilizes a water pervious facing layer for contacting the skin, overlayed by a first fibrous wicking layer, the wicking layer preferably being of the one-way variety in which fluid or liquid is moved away from the facing layer. The product further includes a first container section defined by inner and outer layer of a water pervious wicking material between which is disposed a first absorbent mass. A second container section defined by inner and outer layers between which is disposed a second absorbent mass and a liquid impermeable/gas permeable layer. Spacesuit applications are discussed.

  13. Growth factors with heparin binding affinity in human synovial fluid

    SciTech Connect

    Hamerman, D.; Taylor, S.; Kirschenbaum, I.; Klagsbrun, M.; Raines, E.W.; Ross, R.; Thomas, K.A.

    1987-12-01

    Synovial effusions were obtained from the knees of 15 subjects with joint trauma, menisceal or ligamentous injury, or osteoarthritis. Heparin-Sepharose affinity chromatography of these synovial fluids revealed, in general, three major peaks of mitogenic activity as measured by incorporation of /sup 3/H-thymidine into 3T3 cells. Gradient elution patterns showed activities at 0.5M NaCl, which is characteristic of platelet derived growth factor, and at 1.1 M NaCl and 1.6M NaCl, indicative of acidic and basic fibroblast growth factors, respectively. The identities of these mitogenic fractions were confirmed by specific immunologic and receptor-binding assays. The presence of platelet derived, acidic and basic fibroblast growth factors in the synovial fluid may contribute to wound healing in the arthritic joint.

  14. Acitretin-induced spiny follicular hyperkeratosis.

    PubMed

    Yanik, Mehmet Emin; Erfan, Gamze; Albayrak, Hulya; Tasolar, Kaan; Albayrak, Sule; Gelincik, Ibrahim; Kulac, Mustafa

    2016-06-01

    Spiny follicular hyperkeratosis (SFH) is follicular flesh-colored hyperkeratotic spicules that are linked to different situations including drug reactions. Previously suspected drugs are BRAF inhibitors and cyclosporine. We described a 51-year-old psoriasis patient with SFH who had been using acitretin. PMID:26340416

  15. Rituximab With or Without Yttrium Y-90 Ibritumomab Tiuxetan in Treating Patients With Untreated Follicular Lymphoma

    ClinicalTrials.gov

    2016-06-15

    Stage I Grade 1 Follicular Lymphoma; Stage I Grade 2 Follicular Lymphoma; Stage II Grade 1 Contiguous Follicular Lymphoma; Stage II Grade 1 Non-Contiguous Follicular Lymphoma; Stage II Grade 2 Contiguous Follicular Lymphoma; Stage II Grade 2 Non-Contiguous Follicular Lymphoma; Stage III Grade 1 Follicular Lymphoma; Stage III Grade 2 Follicular Lymphoma; Stage IV Grade 1 Follicular Lymphoma; Stage IV Grade 2 Follicular Lymphoma

  16. Aggregation of Human Eyelid Adipose-derived Stem Cells by Human Body Fluids

    PubMed Central

    Song, Yeonhwa; Yun, Sujin; Yang, Hye Jin; Yoon, A Young; Kim, Haekwon

    2012-01-01

    Fetal bovine serum (FBS) is the most frequently used serum for the cultivation of mammalian cells. However, since animal-derived materials might not be appropriate due to safety issues, allogeneic human serum (HS) has been used to replace FBS, particularly for the culture of human cells. While there has been a debate about the advantages of HS, its precise effect on human adult stem cells have not been clarified. The present study aimed to investigate the effect of HS on the human eyelid adipose stem cells (HEACs) in vitro. When HEACs were cultivated in a medium containing 10% HS, many cells moved into several spots and aggregated there. The phenomenon was observed as early as 9 days following 10% HS treatment, and 12 days following 5% HS plus 5% FBS treatment. However, the aggregation was never observed when the same cells were cultivated with 10% FBS or bovine serum albumin. To examine whether cell density might affect the aggregation, cells were seeded with different densities on 12-well dish. Until the beginning of aggregation, cells seeded at low densities exhibited the longest culture period of 16 days whereas cells seeded at high densities showed the shortest period of 9 days to form aggregation. The number of cells was 15.1±0.2×104 as the least for the low density group, and 29.3±2.8×104 as the greatest for the high density group. When human cord blood serum or normal bovine serum was examined for the same effect on HEACs, interestingly, cord blood serum induced the aggregation of cells whereas bovine serum treatment has never induced. When cells were cultivated with 10% HS for 9 days, they were obtained and analyzed by RT-PCR. Compared to FBS-cultivated HEACs, HS-cultivated HEACs did not express VIM, and less expressed GATA4, PALLD. On the other hand, HS-cultivated HEACs expressed MAP2 more than FBS-cultivated HEACs. In conclusion, human adult stem cells could move and form aggregates by the treatment with human body fluids. PMID:25949109

  17. Follicular lymphoma: evolving therapeutic strategies.

    PubMed

    Kahl, Brad S; Yang, David T

    2016-04-28

    Follicular lymphoma (FL) is the most common indolent non-Hodgkin lymphoma in the Western hemisphere. After decades of stagnation, the natural history of FL appears to have been favorably impacted by the introduction of rituximab. Randomized clinical trials have demonstrated that the addition of rituximab to standard chemotherapy induction has improved the overall survival. Maintenance rituximab strategies can improve progression-free survival. Even chemotherapy platforms have changed in the past 5 years, as bendamustine combined with rituximab has rapidly become a standard frontline strategy in North America and parts of Europe. Recent discoveries have identified patients at high risk for poor outcomes to first-line therapy (m7-Follicular Lymphoma International Prognostic Index [m7-FLIPI]) and for poor outcomes after frontline therapy (National LymphoCare Study). However, several unmet needs remain, including a better ability to identify high-risk patients at diagnosis, the development of predictive biomarkers for targeted agents, and strategies to reduce the risk of transformation. The development of targeted agents, exploiting our current understanding of FL biology, is a high research priority. A multitude of novel therapies are under investigation in both the frontline and relapsed/refractory settings. It will be critical to identify the most appropriate populations for new agents and to develop validated surrogate end points, so that novel agents can be tested (and adopted, if appropriate) efficiently. PMID:26989204

  18. Exposure measurement of aflatoxins and aflatoxin metabolites in human body fluids. A short review.

    PubMed

    Leong, Yin-Hui; Latiff, Aishah A; Ahmad, Nurul Izzah; Rosma, Ahmad

    2012-05-01

    Aflatoxins are highly toxic secondary fungal metabolites mainly produced by Aspergillus flavus and A. parasiticus. Human exposure to aflatoxins may result directly from ingestion of contaminated foods, or indirectly from consumption of foods from animals previously exposed to aflatoxins in feeds. This paper focuses on exposure measurement of aflatoxins and aflatoxin metabolites in various human body fluids. Research on different metabolites present in blood, urine, breast milk, and other human fluids or tissues including their detection techniques is reviewed. The association between dietary intake of aflatoxins and biomarker measurement is also highlighted. Finally, aspects related to the differences between aflatoxin determination in food versus the biomarker approach are discussed. PMID:23606045

  19. Fluid and protein shifts after postural changes in humans

    NASA Technical Reports Server (NTRS)

    Hinghofer-Szalkay, H.; Moser, M.

    1986-01-01

    The effects of head-up, one-step, slow-graded, and rapid-graded tilt positions on blood density (BD), plasma density (PD), and erythrocyte density (ED) in male and female subjects are evaluated using a mass density detection method. The preparation of the samples and procedures for density and hematocrit measurements are described. The formula for calculating the density and volume of fluid shifted between intravascular and extravascular compartments is derived. The data reveal that ED does not change with varying tilt positions; however, PD and BD increase linearly with increasing angles of tilt, thereby resembling the time course of hemodilution and hemoconcentration.

  20. Fluid and protein shifts after postural changes in humans

    SciTech Connect

    Hinghofer-Szalkay, H.; Moser, M.

    1986-01-01

    The effects of head-up, one-step, slow-graded, and rapid-graded tilt positions on blood density (BD), plasma density (PD), and erythrocyte density (ED) in male and female subjects are evaluated using a mass density detection method. The preparation of the samples and procedures for density and hematocrit measurements are described. The formula for calculating the density and volume of fluid shifted between intravascular and extravascular compartments is derived. The data reveal that ED does not change with varying tilt positions; however, PD and BD increase linearly with increasing angles of tilt, thereby resembling the time course of hemodilution and hemoconcentration. 30 references.

  1. Purification and properties of human amniotic fluid diamine oxidase.

    PubMed

    Tufvesson, G

    1978-09-01

    Diamine oxidase (DAO) was purified from amniotic fluid. The activity was separated in two DAO fractions with pI values of 5.8 and 4.0. Molecular weight were found to be 245,000 and 485,000, respectively, with subunit molecular weight of 110,000. This indicated that they probably are dimer and tetramer of the same DAO subunit. The enzyme was active against putrescine and histamine and was strongly inhibited by carbonyl group reagents. A Ping Pong Bi Ter enzyme reaction mechanism is probable. The diamine, with one amino group protonized, is suggested to be responsible for interaction with the enzyme. PMID:30164

  2. Etofenamate levels in human serum and synovial fluid following iontophoresis.

    PubMed

    Bender, T; Bariska, J; Rojkovich, B; Bálint, G

    2001-01-01

    The absorption of etofenamate (CAS 30544-47-9, Rheumon gel) by iontophoresis in 11 patients with low back pain and in 13 patients with synovitis of the knee was evaluated. During the 5-day treatment period, the test gel in a quantity corresponding to 100 mg etofenamate was applied to affected body regions every day by 20-min iontophoresis sessions. Two hours after the fifth application, the concentration of etofenamate in serum and synovial fluid (in patients who had knee joint iontophoresis) were measured by HPLC. Iontophoresis of etofenamate into the lumbar region as well as to the knee joint resulted in consistent serum levels: 219 +/- 136.3 micrograms/l and 191 +/- 84.6 micrograms/l, respectively. In patients with synovitis of the knee, the synovial level of etofenamate (368 +/- 109.2 micrograms/l) was almost twice as high than the serum concentration. The authors conclude that with topical application of etofenamate by iontophoresis the drug appears not only in the serum but also--with higher levels--in the synovial fluid. PMID:11455681

  3. Rituximab and Oblimersen in Treating Patients With Stage II, Stage III, or Stage IV Follicular Non-Hodgkin's Lymphoma

    ClinicalTrials.gov

    2013-01-04

    Stage III Grade 1 Follicular Lymphoma; Stage III Grade 2 Follicular Lymphoma; Stage III Grade 3 Follicular Lymphoma; Stage IV Grade 1 Follicular Lymphoma; Stage IV Grade 2 Follicular Lymphoma; Stage IV Grade 3 Follicular Lymphoma

  4. Numerical solution of fluid-structure interaction represented by human vocal folds in airflow

    NASA Astrophysics Data System (ADS)

    Valášek, J.; Sváček, P.; Horáček, J.

    2016-03-01

    The paper deals with the human vocal folds vibration excited by the fluid flow. The vocal fold is modelled as an elastic body assuming small displacements and therefore linear elasticity theory is used. The viscous incompressible fluid flow is considered. For purpose of numerical solution the arbitrary Lagrangian-Euler method (ALE) is used. The whole problem is solved by the finite element method (FEM) based solver. Results of numerical experiments with different boundary conditions are presented.

  5. Follicular transport route--research progress and future perspectives.

    PubMed

    Knorr, Fanny; Lademann, Jürgen; Patzelt, Alexa; Sterry, Wolfram; Blume-Peytavi, Ulrike; Vogt, Annika

    2009-02-01

    The important role of hair follicles as penetration pathways and reservoir structures for topically applied compounds has been validated in numerous animal models as well as in humans. Follicular penetration rates are modulated by regional variations in size and proportions and the functional status. Advances have especially been made in the targeting of hair follicle-associated cell populations including antigen-presenting cells and stem cells. Improved investigative methods based on differential stripping, spectrophotometry and confocal laser scanning microscopy have led to the determination of the penetration profiles and kinetics for a multiplicity of drugs and drug delivery systems. The observation that particulate delivery systems aggregate and remain in hair follicle openings and their penetration along the follicular duct occurs in a size-dependent manner, which has led to advanced concepts of targeted drug delivery of bioactive compounds in the field of solid particles, as well as semi-solid particles, such as liposomes. This review summarizes the recent progress in this field, and underlines the necessity for pilot studies in human volunteers to further the development of clinical applications for follicular targeting. PMID:19041720

  6. Selective activation of functional suppressor cells by human seminal fluid.

    PubMed Central

    Witkin, S S

    1986-01-01

    The ability of seminal fluid (SF) to induce suppressor cell activity from peripheral blood mononuclear cells (PBMN) was examined. PBMN were incubated with SF for 48 h, washed to remove SF components, treated with mitomycin C (mit C) and co-cultured with Raji cells, a lymphoblastoid cell line. Raji cell proliferation was inhibited by SF-treated PBMN proportionally to SF concentration. SF (50-200 micrograms), mit C-treated Raji cells or mit C-treated PBMN pre-incubated with phytohaemagglutinin were without effect on Raji cell growth. Suppressor T lymphocytes generated by incubation of PBMN with concanavalin A inhibited Raji cells to the same extent as did SF-treated PBMN. All activity was lost following heating at 56 degrees C for 30 min; freezing and thawing reduced the ability of SF to induce suppression by 50%. Dialysis of SF or treatment with antibody to prostaglandin E2 led to a 50% reduction in suppression. PMID:2943541

  7. Sodium imaging of the human knee using soft inversion recovery fluid attenuation

    NASA Astrophysics Data System (ADS)

    Feldman, Rebecca E.; Stobbe, Robert; Watts, Alexander; Beaulieu, Christian

    2013-09-01

    Sodium signal strength in MRI is low when compared with 1H. Thus, image voxel volumes must be relatively large in order to produce a sufficient signal-to-noise ratio (SNR). The measurement of sodium in cartilage is hindered by conflation with signal from the adjacent fluid spaces. Inversion recovery can be used to null signal from fluid, but reduces SNR. The purpose of this work was to optimize inversion recovery sodium MRI to enhance cartilage SNR while nulling fluid. Sodium relaxation was first measured for knee cartilage (T1 = 21 ± 1 ms, T2 fast∗=0.8±0.2 ms, T2 slow∗=19.7±0.5 ms) and fluid (T1 = 48 ± 3 ms, T2∗=47±4 ms) in nine healthy subjects at 4.7 T. The rapid relaxation of cartilage in relation to fluid permits the use of a lengthened inversion pulse to preferentially invert the fluid components. Simulations of inversion pulse length were performed to yield a cartilage SNR enhancing combination of parameters that nulled fluid. The simulations were validated in a phantom and then invivo. B0 inhomogeneity was measured and the effect of off-resonance during the soft inversion pulse was assessed with simulation. Soft inversion recovery yielded twice the SNR and much improved sodium images of cartilage in human knee with little confounding signal from fluid.

  8. Punctate follicular porokeratosis: clinical and pathologic features.

    PubMed

    Trikha, Ritika; Wile, Anna; King, Joy; Ward, Kimberley H M; Brodell, Robert T

    2015-11-01

    Porokeratosis is a disorder of keratinization characterized by an abnormal cornoid lamella surrounding an annular, scaly plaque with an atrophic center. A histologic variant of this condition has been proposed, termed follicular porokeratosis, in cases where follicular involvement was contiguous with an annular cornoid lamella. There has been only 1 report of punctate follicular porokeratosis, in which cornoid lamellae originated exclusively from hair follicles with no associated annular plaque. The authors present the second case of punctate follicular porokeratosis, further supporting the contention that this entity is a unique form of porokeratosis rather than a histologic variant. A 56-year-old African American female presented to the dermatology clinic with a 3-month history of keratotic lesions localized on the right posterior shoulder. Examination revealed an area of perifollicular keratotic papules, each surrounded by an erythematous rim. Histopathology revealed a cornoid lamella originating within a hair follicle, with the parakeratotic column protruding through the follicular orifice. The static nature of the condition along with exclusive involvement of hair follicles supports the notion of punctate follicular porokeratosis as a distinct clinical entity. The diagnosis of this condition relies heavily on proper histopathologic sampling revealing punctate follicular cornoid lamellae. PMID:26485244

  9. FLUID DYNAMICS OF THE HUMAN LARYNX AND UPPER TRACHEOBRONCHIAL AIRWAYS

    EPA Science Inventory

    An airflow pattern that is used to study aerosol deposition in the human lung must attempt to be physiologically realistic. ecause of anatomical conditions, caution must be applied in using only calculated Reynolds number values in biological applications to designate either velo...

  10. Somatostatin reduces sup 3 H-thymidine incorporation and c-myc, but not thyroglobulin ribonucleic acid levels in human thyroid follicular cells in vitro

    SciTech Connect

    degli Uberti, E.C.; Hanau, S.; Rossi, R.; Piva, R.; Margutti, A.; Trasforini, G.; Pansini, G.; del Senno, L. )

    1991-06-01

    The action of somatostatin (SRIH) on {sup 3}H-thymidine (thy) incorporation and on c-myc and thyroglobulin RNA levels in a suspension of follicles from normal and goitrous human thyroid was examined. SRIH, at 10{sup {minus} 7} M concentration, inhibited basal thy incorporation (maximally by 4 h lasting for up 24 h), which effect was greater in goiter than in normal thyroid and was also detected in growing adherent epithelial cells. Moreover, in a follicle suspension SRIH prevented TSH-stimulated thy incorporation, both in normal and in goitrous thyroid. Basal expression of c-myc RNA was not affected by SRIH in either tissue, whereas the TSH-stimulated c-myc RNA level was significantly reduced in goiter. No effect of SRIH was observed on basal or TSH-stimulated thyroglobulin RNA levels. SRIH did not alter basal cAMP concentrations in normal or goitrous follicles, but it significantly reduced TSH-stimulated cAMP accumulation both in normal thyroid and in goiter. Overall, our data indicate a direct inhibitory action of SRIH on growth, but not on differentiation, of human thyroid, probably by a mechanism not entirely cAMP dependent.

  11. Head movement, an important contributor to human cerebrospinal fluid circulation

    PubMed Central

    Xu, Qiang; Yu, Sheng-Bo; Zheng, Nan; Yuan, Xiao-Ying; Chi, Yan-Yan; Liu, Cong; Wang, Xue-Mei; Lin, Xiang-Tao; Sui, Hong-Jin

    2016-01-01

    The suboccipital muscles are connected to the upper cervical spinal dura mater via the myodural bridges (MDBs). Recently, it was suggested that they might work as a pump to provide power for cerebrospinal fluid (CSF) circulation. The purpose of this study was to investigate effects of the suboccipital muscles contractions on the CSF flow. Forty healthy adult volunteers were subjected to cine phase-contrast MR imaging. Each volunteer was scanned twice, once before and once after one-minute-head-rotation period. CSF flow waveform parameters at craniocervical junction were analyzed. The results showed that, after the head rotations, the maximum and average CSF flow rates during ventricular diastole were significantly increased, and the CSF stroke volumes during diastole and during entire cardiac cycle were significantly increased. This suggested that the CSF flow was significantly promoted by head movements. Among the muscles related with head movements, only three suboccipital muscles are connected to the upper cervical spinal dura mater via MDBs. It was believed that MDBs might transform powers of the muscles to CSF. The present results suggested that the head movements served as an important contributor to CSF dynamics and the MDBs might be involved in this mechanism. PMID:27538827

  12. Characterization of lipoproteins in human and canine cerebrospinal fluid (CSF)

    SciTech Connect

    Pitas, R.E.; Weisgraber, K.H.; Boyles, J.K.; Lee, S.; Mahley, R.W.

    1986-03-01

    Previously the authors demonstrated that rat brain astrocytes in vitro synthesize and secrete apo-E and possess apo-B,E(LDL) receptors. The apo-E secreted by astrocytes and apo-E in rat brain extracts differed from serum apo-E in two respects. Brain apo-E had a higher apparent molecular weight and a higher percentage of more acidic isoforms. To characterize further the apo-E within the central nervous system, apo-E in human and canine CSF was investigated. Compared to plasma apo-E, CSF apo-E had a higher apparent M/sub r/ and a higher percentage of acidic isoforms which were sialylated, as shown by neuraminidase digestion. The apo-E in human CSF was approx.5-10% of the plasma level. In CSF 60-80% of the apo-E was in lipoproteins with d = 1.09-1.15. The remainder of the apo-E was in the d > 1.21 fraction. Human CSF lipoproteins were primarily spherical (110-190 A) while canine CSF lipoproteins were a mixture of discs (205 x 65 A) while canine CSF lipoproteins were a mixture of discs (205 x 65 A) and spheres (100-150 A). The CSF also contained apo-AI in the d = 1.09-1.15 g/ml fraction. Human CSF lipoproteins containing both apo-E and apo-AI were isolated on an anti-apo-E affinity column, suggesting that apo-E and AI occurred in the same particles. The CSF apo-E-containing lipoproteins competed for binding of /sup 125/I-LDL to the apo-B,E(LDL) receptor. There was no detectable apo-B in CSF. These data suggest that CSF lipoproteins might transport lipid and regulate lipid homeostasis within the brain.

  13. Fluid volume control during short-term space flight and implications for human performance.

    PubMed

    Watenpaugh, D E

    2001-09-01

    Space flight exerts substantial effects on fluid volume control in humans. Cardiac distension occurs during the first 1-2 days of space flight relative to supine and especially upright 1g conditions. Plasma volume contraction occurs quickly in microgravity, probably as a result of transcapillary fluid filtration into upper-body interstitial spaces. No natriuresis or diuresis has been observed in microgravity, such that diuresis cannot explain microgravity-induced hypovolemia. Reduction of fluid intake occurs irrespective of space motion sickness and leads to hypovolemia. The fourfold elevation of urinary antidiuretic hormone (ADH) levels on flight day 1 probably results from acceleration exposures and other stresses of launch. Nevertheless, it is fascinating that elevated ADH levels and reduced fluid intake occur simultaneously early in flight. Extracellular fluid volume decreases by 10-15% in microgravity, and intracellular fluid volume appears to increase. Total red blood cell mass decreases by approximately 10% within 1 week in space. Inflight Na(+) and volume excretory responses to saline infusion are approximately half those seen in pre-flight supine conditions. Fluid volume acclimation to microgravity sets the central circulation to homeostatic conditions similar to those found in an upright sitting posture on Earth. Fluid loss in space contributes to reduced exercise performance upon return to 1g, although not necessarily in flight. In-flight exercise training may help prevent microgravity-induced losses of fluid and, therefore, preserve the capacity for upright exercise post-flight. Protection of orthostatic tolerance during space flight probably requires stimulation of orthostatic blood pressure control systems in addition to fluid maintenance or replacement. PMID:11581336

  14. Follicular contact dermatitis revisited: A review emphasizing neomycin-associated follicular contact dermatitis

    PubMed Central

    Cohen, Philip R

    2014-01-01

    Follicular contact dermatitis clinically presents as individual papules that include a central hair follicle. Pathologic features involve the follicle and the surrounding dermis: spongiosis and vesicle formation of the follicular epithelium associated with perifollicular and perivascular lymphocytic inflammation. Using the PubMed database, an extensive literature search was performed on follicular contact dermatitis and neomycin. Relevant papers were reviewed and the clinical and pathologic features, the associated chemicals (including a more detailed description of neomycin), the hypothesized pathogenesis, and the management of follicular contact dermatitis were described. Several agents-either as allergens or irritants-have been reported to elicit follicular contact dermatitis. Several hypotheses have been suggested for the selective involvement of the follicles in follicular contact dermatitis: patient allergenicity, characteristics of the agent, vehicle containing the agent, application of the agent, and external factors. The differential diagnosis of follicular contact dermatitis includes not only recurrent infundibulofolliculitis, but also drug eruption, mite infestation, viral infection, and dermatoses that affect hair follicles. The primary therapeutic intervention for follicular contact dermatitis is withdrawal of the causative agent; treatment with a topical corticosteroid preparation may also promote resolution of the dermatitis. In conclusion, follicular contact dermatitis may be secondary to allergens or irritants; topical antibiotics, including neomycin, may cause this condition. Several factors may account for the selective involvement of the hair follicle in this condition. Treatment of the dermatitis requires withdrawal of the associated topical agent; in addition, topical corticosteroids may be helpful to promote resolution of lesions. PMID:25516854

  15. Fluid Secretion in Isolated Proximal Straight Renal Tubules EFFECT OF HUMAN UREMIC SERUM

    PubMed Central

    Grantham, Jared J.; Irwin, Richard L.; Qualizza, Patti B.; Tucker, Donald R.; Whittier, Frederick C.

    1973-01-01

    We have examined the effect of normal and uremic human sera on the transtubular flow of fluid in isolated perfused segments of rabbit proximal convoluted and straight renal tubules. Proximal convoluted and straight tubules absorbed fluid from the lumen when the external bath was normal rabbit serum. Normal human sera in the bath depressed net fluid absorption in both tubular segments, but more importantly, uremic human serum caused proximal straight tubules to secrete fluid into the lumen. Fluid secretion was also demonstrated indirectly by observing in nonperfused proximal straight, but not proximal convoluted tubules, that the normally collapsed lumens opened widely in uremic serum. Nonperfused proximal straight tubules developed expanded lumens even after a 25-fold dilution of human uremic serum with normal rabbit serum, whereas lumen expansion occurred only in undiluted normal human serum, on the average. Serum from acutely uremic rabbits possessed secretory activity but normal rabbit serum did not. The secretory effect of uremic sera in proximal straight tubules was inhibited by cooling and ouabain and probenecid. The secretory activity of uremic sera was removed by dialysis, but not by freezing or boiling. Para-aminohippurate and benzoate caused fluid secretion in proximal straight tubules but urea, creatinine, guanidinosuccinate, and urate did not. On the basis of these results, we suggest that the secretory factor in serum may be a substance or group of substances possibly related to the hippurate class of organic molecules that are accumulated to relatively high concentrations in renal failure. The secretory material in the serum of uremic patients may significantly influence the transport of salt and water in relatively intact residual nephrons. Images PMID:4738063

  16. Primate follicular development and oocyte maturation in vitro

    PubMed Central

    Xu, Jing; Xu, Min; Bernuci, Marcelo P; Fisher, Thomas E; Shea, Lonnie D; Woodruff, Teresa K; Zelinski, Mary B; Stouffer, Richard L

    2014-01-01

    The factors and processes involved in primate follicular development are complex and not fully understood. An encapsulated three-dimensional (3D) follicle culture system could be a valuable in vitro model to study the dynamics and regulation of folliculogenesis in intact individual follicles in primates. Besides the research relevance, in vitro follicle maturation (IFM) is emerging as a promising approach to offer options for fertility preservation in female patients with cancer. This review summarizes the current published data on in vitro follicular development from the preantral to small antral stage in nonhuman primates, including follicle survival and growth, endocrine (ovarian steroid hormone) and paracrine/autocrine (local factor) function, as well as oocyte maturation and fertilization. Future directions include major challenges and strategies to further improve follicular growth and differentiation with oocytes competent for in vitro fertilization and subsequent embryonic development, as well as opportunities to investigate primate folliculogenesis by utilizing this 3D culture system. The information may be valuable in identifying optimal conditions for human follicle culture, with the ultimate goal of translating the experimental results and products to patients, thereby facilitating diagnostic and therapeutic approaches for female fertility. PMID:24097381

  17. Impact of follicular aspiration on folliculogenesis as influenced by dietary energy and stage of lactation.

    PubMed

    Gwazdauskas, F C; Kendrick, K W; Pryor, A W; Bailey, T L

    2000-07-01

    Prolonged dietary energy restriction has been shown to influence the secretion of ovarian hormones and follicular development. Energy balance affects the ability of the cow to support a functional reproductive system during early lactation. Follicular aspiration is a tool that can be used to assess the impact of dietary energy and stage of lactation on follicle development in dairy cattle. Follicular aspiration was conducted twice weekly from d 30 to 100 postpartum in cows fed either 104 or 80% of NRC energy requirements. Oocyte numbers recovered increased linearly from d 30 to 100 postpartum. Cows on high energy diets produced more high quality oocytes, but also produced more poor quality oocytes than cows fed the lower energy diet. Follicles < 5 mm predominated throughout. Cows in early (d 28) and mid (d 117) lactation were used to assess stage of lactation effects on serum and follicular fluid hormones, and quality of oocytes recovered by follicular aspiration procedures for 10 wk. Serum IGF-1 decreased in early lactation cows, but increased in midlactation cows. Follicular fluid progesterone in early lactation cows decreased from 275 to 235 ng/ml, whereas in midlactation cows it increased from 65 to 550 ng/ml at aspiration wk 10. The percentage of high quality ova was higher in early lactation cows (11%) than midlactation cows (6%), but fell to 1% by aspiration wk 7 and increased to 10% by wk 10, whereas percentage of high quality ova in midlactation cows steadily declined to 2% by wk 10. Milk yield and energy balance affect patterns of folliculogenesis in lactating dairy cows. PMID:10908066

  18. Rituximab, Lenalidomide, and Ibrutinib in Treating Patients With Previously Untreated Stage II-IV Follicular Lymphoma

    ClinicalTrials.gov

    2016-08-24

    Stage II Grade 1 Contiguous Follicular Lymphoma; Stage II Grade 1 Non-Contiguous Follicular Lymphoma; Stage II Grade 2 Contiguous Follicular Lymphoma; Stage II Grade 2 Non-Contiguous Follicular Lymphoma; Stage II Grade 3 Contiguous Follicular Lymphoma; Stage II Grade 3 Non-Contiguous Follicular Lymphoma; Stage III Grade 1 Follicular Lymphoma; Stage III Grade 2 Follicular Lymphoma; Stage III Grade 3 Follicular Lymphoma; Stage IV Grade 1 Follicular Lymphoma; Stage IV Grade 2 Follicular Lymphoma; Stage IV Grade 3 Follicular Lymphoma

  19. Isoflavones in human breast milk and other biological fluids.

    PubMed

    Franke, A A; Custer, L J; Tanaka, Y

    1998-12-01

    We established a method for using HPLC and diode-array ultraviolet scanning to quantitate soy isoflavonoids in foods and in human plasma, urine, and breast milk. The analytes occurring as glycoside conjugates were hydrolyzed enzymatically before HPLC analysis if extracted from biological matrices or were subjected to direct HPLC analysis after extraction from foods. We monitored the isoflavones daidzein, genistein, glycitein, formononetin, and biochanin-A and their mammalian metabolites equol and O-desmethylangolensin in human plasma, urine, and breast milk. Analytes were identified by absorbance patterns, fluorometric and electrochemical detection. and comparison with internal and external standards. In addition, we identified analytes by using gas chromatography-mass spectrometry after trimethylsilylation. The HPLC method was also used to measure concentrations of isoflavones and their glucoside conjugates in various soy-based infant formulas. Total isoflavone concentrations varied between 155 and 281 mg/kg. After one woman received a moderate challenge with 20 g roasted soybeans (equivalent to 37 mg isoflavones), we detected mean total isoflavone concentrations of approximately 2.0 micromol/L in plasma, 0.2 micromol/L in breast milk, and 3.0 micromol/h in urine. According to our measurements, with adjustment for body weight, isoflavonoid exposure is 4-6 times higher in infants fed soy-based formula than in adults eating a diet rich in soyfoods (approximately 30 g/d). Implications of the presented results for the potential cancer-preventing activity of isoflavones by exposing newborn infants to these phytochemicals are discussed. PMID:9848518

  20. Fluid and electrolyte control systems in the human body: A study report

    NASA Technical Reports Server (NTRS)

    White, R. J.

    1973-01-01

    Research in the area of modeling of the fluid and electrolyte system is briefly reviewed and a model of this system, which is adequate for a basic description of the requisite physiological processes, is presented. The use of this model as an individual subsystem model and as a component of a more complete human model is discussed.

  1. Experiments on the fluid dynamics of the human cough

    NASA Astrophysics Data System (ADS)

    Settles, Gary

    2011-11-01

    Human coughing is studied non-intrusively by high-speed schlieren videography, revealing a turbulent jet lasting up to 1 sec with a total expelled air volume of about 2 L. Velocimetry of eddy motion reveals a jet centerline airspeed of at least 8 m/sec. With Re roughly 18,000 the cough jet is inertia-driven and buoyancy is negligible. It shows typical round-turbulent-jet behavior, including a conical spreading angle of 24 deg, despite irregular initial conditions. The cough jet is projected several m into the surrounding air before it mixes out. It is well known that a cough can transmit infectious agents, and we are advised to cover our mouths in an apparent attempt to thwart the jet formation. Present experiments have shown that wearing a surgical mask or respirator designed to prevent the inhalation of infectious agents also interferes with the cough-jet formation, redirecting it into the person's rising thermal plume. (Tang et al., J. Royal. Soc. Interface 6, S727, 2009.)

  2. Detection of Human Herpesvirus 6 and Varicella-Zoster Virus in Tear Fluid of Patients with Bell's Palsy by PCR

    PubMed Central

    Pitkäranta, A.; Piiparinen, H.; Mannonen, L.; Vesaluoma, M.; Vaheri, A.

    2000-01-01

    Human herpesvirus 6 DNA was detected by PCR in the tear fluid of 7 (35%) of 20 patients with Bell's palsy and of 1 (5%) of 20 healthy controls. Varicella-zoster virus was detected by PCR in the tear fluid of 2 of 20 Bell's palsy patients but in none of the tear fluids from 20 healthy controls. These findings suggest an association between human herpesviruses and Bell's palsy. PMID:10878079

  3. Genetics of follicular lymphoma transformation

    PubMed Central

    Pasqualucci, Laura; Khiabanian, Hossein; Fangazio, Marco; Vasishtha, Mansi; Messina, Monica; Holmes, Antony B.; Ouillette, Peter; Trifonov, Vladimir; Rossi, Davide; Tabbò, Fabrizio; Ponzoni, Maurilio; Chadburn, Amy; Murty, Vundavalli V.; Bhagat, Govind; Gaidano, Gianluca; Inghirami, Giorgio; Malek, Sami N.; Rabadan, Raul; Dalla-Favera, Riccardo

    2014-01-01

    Summary Follicular lymphoma (FL) is an indolent disease, but 30-40% of cases undergo histologic transformation to an aggressive malignancy, typically represented by diffuse large B cell lymphoma (DLBCL). The pathogenesis of this process remains largely unknown. Using whole-exome sequencing and copy-number analysis, here we show that the dominant clone of FL and transformed FL (tFL) arise by divergent evolution from a common mutated precursor through the acquisition of distinct genetic events. Mutations in epigenetic modifiers and anti-apoptotic genes are introduced early in the common precursor, while tFL is specifically associated with alterations deregulating cell-cycle progression and DNA-damage responses (CDKN2A/B, MYC, TP53), as well as with aberrant somatic hypermutation. The genomic profile of tFL shares similarities with that of germinal center B-cell-type de novo DLBCL, but also displays unique combinations of altered genes, with diagnostic and therapeutic implications. PMID:24388756

  4. Workshop on Critical Issues in Microgravity Fluids, Transport, and Reaction Processes in Advanced Human Support Technology

    NASA Technical Reports Server (NTRS)

    Chiaramonte, Francis P.; Joshi, Jitendra A.

    2004-01-01

    This workshop was designed to bring the experts from the Advanced Human Support Technologies communities together to identify the most pressing and fruitful areas of research where success hinges on collaborative research between the two communities. Thus an effort was made to bring together experts in both advanced human support technologies and microgravity fluids, transport and reaction processes. Expertise was drawn from academia, national laboratories, and the federal government. The intent was to bring about a thorough exchange of ideas and develop recommendations to address the significant open design and operation issues for human support systems that are affected by fluid physics, transport and reaction processes. This report provides a summary of key discussions, findings, and recommendations.

  5. In Vitro Studies Evaluating Leaching of Mercury from Mine Waste Calcine Using Simulated Human Body Fluids

    PubMed Central

    2010-01-01

    In vitro bioaccessibility (IVBA) studies were carried out on samples of mercury (Hg) mine-waste calcine (roasted Hg ore) by leaching with simulated human body fluids. The objective was to estimate potential human exposure to Hg due to inhalation of airborne calcine particulates and hand-to-mouth ingestion of Hg-bearing calcines. Mine waste calcines collected from Hg mines at Almadén, Spain, and Terlingua, Texas, contain Hg sulfide, elemental Hg, and soluble Hg compounds, which constitute primary ore or compounds formed during Hg retorting. Elevated leachate Hg concentrations were found during calcine leaching using a simulated gastric fluid (as much as 6200 μg of Hg leached/g sample). Elevated Hg concentrations were also found in calcine leachates using a simulated lung fluid (as much as 9200 μg of Hg leached/g), serum-based fluid (as much as 1600 μg of Hg leached/g), and water of pH 5 (as much as 880 μg of Hg leached/g). The leaching capacity of Hg is controlled by calcine mineralogy; thus, calcines containing soluble Hg compounds contain higher leachate Hg concentrations. Results indicate that ingestion or inhalation of Hg mine-waste calcine may lead to increased Hg concentrations in the human body, especially through the ingestion pathway. PMID:20491469

  6. T follicular helper (Tfh ) cells in normal immune responses and in allergic disorders.

    PubMed

    Varricchi, G; Harker, J; Borriello, F; Marone, G; Durham, S R; Shamji, M H

    2016-08-01

    Follicular helper T cells (Tfh ) are located within germinal centers of lymph nodes. Cognate interaction between Tfh , B cells, and IL-21 drives B cells to proliferate and differentiate into plasma cells thereby leading to antibody production. Tfh cells and IL-21 are involved in infectious and autoimmune diseases, immunodeficiencies, vaccination, and cancer. Human peripheral blood CXCR5(+) CD4(+) T cells comprise different subsets of Tfh -like cells. Despite the importance of the IgE response in the pathogenesis of allergic disorders, little is known about the role of follicular and blood Tfh cells and IL-21 in human and experimental allergic disease. Here, we review recent advances regarding the phenotypic and functional characteristics of both follicular and blood Tfh cells and of the IL-21/IL-21R system in the context of allergic disorders. PMID:26970097

  7. Phenotypes of the ovarian follicular basal lamina predict developmental competence of oocytes

    PubMed Central

    Irving-Rodgers, Helen F.; Morris, Stephanie; Collett, Rachael A.; Peura, Teija T.; Davy, Margaret; Thompson, Jeremy G.; Mason, Helen D.; Rodgers, Raymond J.

    2009-01-01

    BACKGROUND The ovarian follicular basal lamina underlies the epithelial membrana granulosa and maintains the avascular intra-follicular compartment. Additional layers of basal lamina occur in a number of pathologies, including pili annulati and diabetes. We previously found additional layers of follicular basal lamina in a significant percentage of healthy bovine follicles. We wished to determine if this phenomenon existed in humans, and if it was related to oocyte function in the bovine. METHODS AND RESULTS We examined follicles from human ovaries (n = 18) by electron microscopy and found that many follicles had additional layers of basal lamina. Oocytes (n = 222) from bovine follicles with normal or unusual basal laminas were isolated and their ability to undergo in vitro maturation, fertilization and culture to blastocyst was compared. Healthy bovine follicles with a single layer of basal lamina had oocytes with significantly (P < 0.01) greater developmental competence than healthy follicles with additional layers of follicular basal lamina (65% versus 28%). CONCLUSIONS These findings provide direct evidence that the phenotype of the follicular basal lamina is related to oocyte competence. PMID:19095662

  8. Predictive value of bovine follicular components as markers of oocyte developmental potential.

    PubMed

    Matoba, Satoko; Bender, Katrin; Fahey, Alan G; Mamo, Solomon; Brennan, Lorraine; Lonergan, Patrick; Fair, Trudee

    2014-01-01

    The follicle is a unique micro-environment within which the oocyte can develop and mature to a fertilisable gamete. The aim of this study was to investigate the ability of a panel of follicular parameters, including intrafollicular steroid and metabolomic profiles and theca, granulosa and cumulus cell candidate gene mRNA abundance, to predict the potential of bovine oocytes to develop to the blastocyst stage in vitro. Individual follicles were dissected from abattoir ovaries, carefully ruptured under a stereomicroscope and the oocyte was recovered and individually processed through in vitro maturation, fertilisation and culture. The mean (±s.e.m.) follicular concentrations of testosterone (62.8±4.8 ngmL(-1)), progesterone (616.8±31.9 ngmL(-1)) and oestradiol (14.4±2.4 ngmL(-1)) were not different (P>0.05) between oocytes that formed (competent) or failed to form (incompetent) blastocysts. Principal-component analysis of the quantified aqueous metabolites in follicular fluid showed differences between oocytes that formed blastocysts and oocytes that degenerated; l-alanine, glycine and l-glutamate were positively correlated and urea was negatively correlated with blastocyst formation. Follicular fluid associated with competent oocytes was significantly lower in palmitic acid (P=0.023) and total fatty acids (P=0.031) and significantly higher in linolenic acid (P=0.036) than follicular fluid from incompetent oocytes. Significantly higher (P<0.05) transcript abundance of LHCGR in granulosa cells, ESR1 and VCAN in thecal cells and TNFAIP6 in cumulus cells was associated with competent compared with incompetent oocytes. PMID:23514964

  9. Roles of extracellular matrix in follicular development.

    PubMed

    Rodgers, R J; van Wezel, I L; Irving-Rodgers, H F; Lavranos, T C; Irvine, C M; Krupa, M

    1999-01-01

    The cellular biology and changes in the extracellular matrix of ovarian follicles during their development are reviewed. During growth of the bovine ovarian follicle the follicular basal lamina doubles 19 times in surface area. It changes in composition, having collagen IV alpha 1-26 and laminin alpha 1, beta 2 and gamma 1 at the primordial stage, and collagen IV alpha 1 and alpha 2, reduced amounts of alpha 3-alpha 5, and a higher content of laminin alpha 1, beta 2 and gamma 1 at the antral stage. In atretic antral follicles laminin alpha 2 was also detected. The follicular epithelium also changes from one layer to many layers during follicular growth. It is clear that not all granulosal cells have equal potential to divide, and we have evidence that the granulosal cells arise from a population of stem cells. This finding has important ramifications and supports the concept that different follicular growth factors can act on different subsets of granulosal cells. In antral follicles, the replication of cells occurs in the middle layers of the membrana granulosa, with older granulosal cells towards the antrum and towards the basal lamina. The basal cells in the membrana granulosa have also been observed to vary in shape between follicies. In smaller antral follicles, they were either columnar or rounded, and in follicles > 5 mm the cells were all rounded. The reasons for these changes in matrix and cell shapes are discussed in relation to follicular development. PMID:10692866

  10. Fluid shifts and muscle function in humans during acute simulated weightlessness

    NASA Technical Reports Server (NTRS)

    Hargens, A. R.; Tipton, C. M.; Gollnick, P. D.; Mubarak, S. J.; Tucker, B. J.; Akeson, W. H.

    1983-01-01

    The acute effects of simulated weightlessness on transcapillary fluid balance, tissue fluid shifts, muscle function, and triceps surface reflex time were studied in eight supine human subjects who were placed in a 5 degrees head-down tilt position for 8 hr. Results show a cephalic fluid shift from the legs as indicated by facial edema, nasal congestion, increased urine flow, decreased creatinine excretion, reduced calf girth, and decreased lower leg volume. The interstitial fluid pressure in the tibialis anterior muscle and subcutaneous tissue of the lower leg was found to fall significantly, while other transcapillary pressures (capillary and interstitial fluid colloid osmotic pressures) were relatively unchanged. The total water content of the soleus muscle was unchanged during the head-down tilt. After head-down tilt, isometric strength and isokinetic strength of the plantar flexors were unchanged, while the triceps surae reflex time associated with plantar flexion movement slowed slightly. These results demonstrate a dehydration effect of head-down tilt on muscle and subcutaneous tissue of the lower leg that may affect muscle function.