Milk progesterone as a tool to improve fertility in dairy cows

Milk progesterone offers an opportunity to objectively study fertility in dairy cows, in contrast to traditional measures of dairy cow fertility, which in general are highly influenced by on-farm management decisions. The aim of this thesis was to study how milk progesterone could be used as a genetic and management tool to improve fertility in dairy cows. Progesterone-based measures were influenced by different systematic factors, e.g. breed, parity, season, housing and lameness, studied in a dataset from a Swedish experimental herd. The repeatabilities were higher for progesterone-based measures compared with traditional measures of fertility based on insemination data. If a cow had an atypical progesterone profile in one lactation, the risk of an atypical profile in the next lactation was increased. Genetic parameters for progesterone measures based on different milk sampling intervals were estimated in a British dataset. Heritability estimates were moderate, but decreased with increased sampling intervals. It was shown that progesterone analysis of monthly milk samples, resembling milk sampling as in the current Swedish milk recording system, could be used to increase the accuracy of genetic evaluation for an earlier start of cyclical ovarian activity after calving. Inclusion of monthly milk sampling for progesterone analysis in predictive models could also be used to identify cows with delayed ovarian cyclicity with a high accuracy already two months after calving. This enables an earlier treatment of ovarian dysfunction and therefore, probably, a shorter calving interval. In conclusion, this thesis shows that milk progesterone may be used for improved management and genetic evaluation of dairy cow fertility

A nongenomic mechanism for progesterone-mediated immunosuppression: Inhibition of K+ channels, Ca2+ signaling, and gene expression in T lymphocytes

The mechanism by which progesterone causes localized suppression of the immune response during pregnancy has remained elusive. Using human T lymphocytes and T cell lines, we show that progesterone, at concentrations found in the placenta, rapidly and reversibly blocks voltage-gated and calcium-activated K+ channels (KV and KCa, respectively), resulting in depolarization of the membrane potential. As a result, Ca2+ signaling and nuclear factor of activated T cells (NF-AT)-driven gene expression are inhibited. Progesterone acts distally to the initial steps of T cell receptor (TCR)-mediated signal transduction, since it blocks sustained Ca2+ signals after thapsigargin stimulation, as well as oscillatory Ca2+ signals, but not the Ca2+ transient after TCR stimulation. K+ channel blockade by progesterone is specific; other steroid hormones had little or no effect, although the progesterone antagonist RU 486 also blocked KV and KCa channels. Progesterone effectively blocked a broad spectrum of K+ channels, reducing both Kv1.3 and charybdotoxin-resistant components of KV current and KCa current in T cells, as well as blocking several cloned KV channels expressed in cell lines. Progesterone had little or no effect on a cloned voltage-gated Na+ channel, an inward rectifier K+ channel, or on lymphocyte Ca2+ and Cl- channels. We propose that direct inhibition of K+ channels in T cells by progesterone contributes to progesterone-induced immunosuppression

A nongenomic mechanism for progesterone-mediated immunosuppression: Inhibition of K+ channels, Ca2+ signaling, and gene expression in T lymphocytes

The mechanism by which progesterone causes localized suppression of the immune response during pregnancy has remained elusive. Using human T lymphocytes and T cell lines, we show that progesterone, at concentrations found in the placenta, rapidly and reversibly blocks voltage-gated and calcium-activated K+ channels (KV and KCa, respectively), resulting in depolarization of the membrane potential. As a result, Ca2+ signaling and nuclear factor of activated T cells (NF-AT)-driven gene expression are inhibited. Progesterone acts distally to the initial steps of T cell receptor (TCR)-mediated signal transduction, since it blocks sustained Ca2+ signals after thapsigargin stimulation, as well as oscillatory Ca2+ signals, but not the Ca2+ transient after TCR stimulation. K+ channel blockade by progesterone is specific; other steroid hormones had little or no effect, although the progesterone antagonist RU 486 also blocked KV and KCa channels. Progesterone effectively blocked a broad spectrum of K+ channels, reducing both Kv1.3 and charybdotoxin-resistant components of KV current and KCa current in T cells, as well as blocking several cloned KV channels expressed in cell lines. Progesterone had little or no effect on a cloned voltage-gated Na+ channel, an inward rectifier K+ channel, or on lymphocyte Ca2+ and Cl- channels. We propose that direct inhibition of K+ channels in T cells by progesterone contributes to progesterone-induced immunosuppression

Single-cell analysis of [Ca²⁺]i signalling in sub-fertile men:characteristics and relation to fertilization outcome

STUDY QUESTIONWhat are the characteristics of progesterone-induced (CatSper-mediated) single cell [Ca2+]i signals in spermatozoa from sub-fertile men and how do they relate to fertilizing ability?SUMMARY ANSWERSingle cell analysis of progesterone-induced (CatSper-mediated) [Ca2+]i showed that reduced progesterone-sensitivity is a common feature of sperm from sub-fertile patients and is correlated with fertilization rate.WHAT IS KNOWN ALREADYStimulation with progesterone is a widely used method for assessing [Ca2+]i mobilization by activation of CatSper in human spermatozoa. Although data are limited, sperm population studies have indicated an association of poor [Ca2+]i response to progesterone with reduced fertilization ability.STUDY DESIGN, SIZE, DURATIONThis was a cohort study using semen samples from 21 donors and 101 patients attending the assisted conception unit at Ninewells Hospital Dundee who were undergoing ART treatment. Patients were recruited from January 2016 to June 2017.PARTICIPANTS/MATERIALS, SETTING, METHODSSemen donors and patients were recruited in accordance with local ethics approval (13/ES/0091) from the East of Scotland Research Ethics Service (EoSRES) REC1. [Ca2+]i responses were examined by single cell imaging and motility parameters assessed by computer-assisted sperm analysis (CASA).MAIN RESULTS AND THE ROLE OF CHANCEFor analysis, patient samples were divided into three groups IVF(+ve) (successful fertilization; 62 samples), IVF-FF (failed fertilization; eight samples) and ICSI (21 samples). A further 10 IVF samples showed large, spontaneous [Ca2+]i oscillations and responses to progesterone could not be analysed. All patient samples loaded with the [Ca2+]i-indicator fluo4 responded to progesterone stimulation with a biphasic increase in fluorescence (transient followed by plateau) which resembled that seen in progesterone-stimulated donor samples. The mean normalized response (progesterone-induced increase in fluorescence normalized to resting level) was significantly smaller in IVF-FF and ICSI patient groups than in donors. All samples were further analysed by plotting, for each cell, the relationship between resting fluorescence intensity and the progesterone-induced fluorescence increment. In donor samples these plots overlaid closely and had a gradient of ≈ 2 and plots for most IVF(+ve) samples closely resembled the donor distribution. However, in a subset (≈ 10%) of IVF(+ve) samples, 3/8 IVF-FF samples and one-third of ICSI samples the gradient of the plot was significantly lower, indicating that the response to progesterone of the cells in these samples was abnormally small. Examination of the relationship between gradient (regression coefficient of the plot) in IVF samples and fertilization rate showed a positive correlation. In IVF-FF and ICSI groups, the proportion of cells in which a response to progesterone could be detected was significantly lower than in donors and IVF (+ve) patients. Approximately 20% of cells in donor, IVF(+ve) and ICSI samples generated [Ca2+]i oscillations when challenged with progesterone but in IVF-FF samples only ≈ 10% of cells generated oscillations and there was a significantly greater proportion of samples where no oscillations were observed. Levels of hyperactivated motility were lower in IVF(+ve) and IVF-FF groups compared to controls, IVF-FF also having lower levels than IVF(+ve).LIMITATIONS, REASONS FOR CAUTIONThis is an in vitro study and caution must be taken when extrapolating these results in vivo.WIDER IMPLICATIONS OF THE FINDINGSThis study reveals important details of impaired [Ca2+]i signalling in sperm from sub-fertile men that cannot be detected in population studies

Crucial cross-talk of interleukin-1β and progesterone in human choriocarcinoma

Copyright @ 2012 Spandidos Publications Ltd. This article can be accessed from the links below.This article has been made available through the Brunel Open Access Publishing Fund.Choriocarcinoma is a highly malignant epithelial tumour that is most often associated with hydatidiform mole and presents the most common emergency medical problem in the management of trophoblast disease. We hypothesise that the hormones/cytokines present within the tumour microenvironment play key roles in the development of choriocarcinoma. In this study we assessed the effects of interleukin-1β (IL-1β) on cell death in the presence or absence of the sex hormone progesterone using two choriocarcinoma cell lines (BeWo and JEG-3) as in vitro experimental models. Although IL-1β induced cell death in both cell lines, the effect was more pronounced in JEG-3 cells, where cell death reached 40% compared to 15% in BeWo cells. Cell death of JEG-3 cells in response to IL-1β was significantly decreased by co-treatment with 100 nM and 1000 nM progesterone and completely abolished at a progesterone concentration of 1000 nM. Progesterone was also able to induce phosphorylation of ERK1/2 in these cells. Pretreatment of JEG-3 cells with a specific MAPK inhibitor (UO126) inhibited progesterone's inhibitory effect on cell death. Collectively, these data provide evidence of cross-talk between progesterone and IL-1β in this aggressive and poorly understood tumour that involves activation of a MAPK pathway and involvement of numerous progesterone receptors.This research was funded by a National Institutes of Health Grant ESO12961. This article is made available through the Brunel Open Access Publishing Fund

Human amniotic fluid glycoproteins expressing sialyl Lewis carbohydrate antigens stimulate progesterone production in human trophoblasts in vitro

Background: Progesterone is thought to mediate immune modulator effects by regulating uterine responsiveness. The aim of the study was to clarify the effect of transferrin and glycodelin A (former name PP14) as sialyl Lewis X-expressing glycoproteins on the release of progesterone by trophoblast cells in vitro. Methods: Cytotrophoblast cells were prepared from human term placentas by standard dispersion of villous tissue followed by a Percoll gradient centrifugation step. Trophoblasts were incubated with varying concentrations (50-300 mug/ml) of human amniotic fluid- and serum-transferrin as well as with glycodelin A. Culture supernatants were assayed for progesterone, human chorionic gonadotropin (hCG) and cortisol by enzyme immunometric methods. Results: The release of progesterone is increased in amniotic fluid transferrin- and glycodelin A-treated trophoblast cell cultures compared to untreated trophoblast cells. There is no relation between transferrin and the hCG or cortisol production of trophoblast cells. Conclusion: The results suggest that sialyl Lewis carbohydrate antigen-expressing amniotic fluid glycoproteins modulate the endocrine function of trophoblasts in culture by upregulating progesterone production. Copyright (C) 2004 S. Karger AG, Basel

Trial protocol OPPTIMUM : does progesterone prophylaxis for the prevention of preterm labour improve outcome?

Background Preterm birth is a global problem, with a prevalence of 8 to 12% depending on location. Several large trials and systematic reviews have shown progestogens to be effective in preventing or delaying preterm birth in selected high risk women with a singleton pregnancy (including those with a short cervix or previous preterm birth). Although an improvement in short term neonatal outcomes has been shown in some trials these have not consistently been confirmed in meta-analyses. Additionally data on longer term outcomes is limited to a single trial where no difference in outcomes was demonstrated at four years of age of the child, despite those in the “progesterone” group having a lower incidence of preterm birth. Methods/Design The OPPTIMUM study is a double blind randomized placebo controlled trial to determine whether progesterone prophylaxis to prevent preterm birth has long term neonatal or infant benefit. Specifically it will study whether, in women with singleton pregnancy and at high risk of preterm labour, prophylactic vaginal natural progesterone, 200 mg daily from 22 – 34 weeks gestation, compared to placebo, improves obstetric outcome by lengthening pregnancy thus reducing the incidence of preterm delivery (before 34 weeks), improves neonatal outcome by reducing a composite of death and major morbidity, and leads to improved childhood cognitive and neurosensory outcomes at two years of age. Recruitment began in 2009 and is scheduled to close in Spring 2013. As of May 2012, over 800 women had been randomized in 60 sites. Discussion OPPTIMUM will provide further evidence on the effectiveness of vaginal progesterone for prevention of preterm birth and improvement of neonatal outcomes in selected groups of women with singleton pregnancy at high risk of preterm birth. Additionally it will determine whether any reduction in the incidence of preterm birth is accompanied by improved childhood outcome

إنتاج مادة 17 ألفا هيدروكسي البروجستيرون على مستوى المخمر المعملي بواسطة فطرة كاننجهاميلا إيكينولاتا

The mircrobiological transformation of progesterone by a local isolate of Cunninghamella echiiiulata using a laboratory fermentor was studied. Progresterone (10-50 g/1) wetted by Tween 80 was added to 48-hour old culture and the transformation was left to proceed for 72 hours. Thereafter, the different transformation products were resolved chromatog-raphically. The identity of each product was established through the determination of m.p., mixed m.p., optical rotation and ultraviolet as well as infrared absorption spectra. A comparison of the R{ values of each product with that of the corresponding reference using different solvent systems as well as their colour expressed with two spray reagents, was used as a further proof for the identity of the isolated products. With all concentrations of progesterone tested, maximum yield of 17ot -hydroxyprogesterone was obtained after 48 hours of fermentation Progesterone concentrations of 10 and 20 g/1 were almost quantitatively converted to the different transformation products after 72 hours of fermentation. Using a concentration of 20 g/1 and incubation period of 48 hours, the transformation product mixture consisted of unchanged progesterone (6%), 17 o< -hydroxyprogesterone (54%),llotrhydroxyprogesterone (29%) and llo<;,17<^-dihydroxy-progesterone (2.5%).تم استخدام مخمر صناعي سعة 2 لتر لاختيار مقدرة الفطرة على تكوين هذه المادة في ظروف تشبه تلك المطبقة في الصناعة . وبدراسة تركيزات متعددة فن مادة البروجستيرون تتراوح ما بين 10جرام /لتر إلى 50جرام /لتر ، وجد أن أنسب التركيزات المختبرة هو تركيز 20 جرام من البروجستيرون لكل لترمن الوسط الغذائي ، حيث تم تحويل كل البووجستيرون المضاف إلى المشتقات المختلفة خلال 72 ساعة من بدء الاضافة . ووجد أن أعلى معدل لتكوين مادة 17 ألفا - هيدروكسي البروجستيرون كان بعد 48 ساعة من بدأ إضافة البروجستيرون . عند فصل المواد الناتجة من تحول البروجستيرون بواسطة الفطرة المستخدمة وذلك بواسطة أعمدة الفصل باستخدام مادة الالومينا وجد أن البروجستيرون يتحول إلى : 17 ألفا - هيدروكسي البروجستيرون ( 54 %) 11 ألفا - هيدروكسي البروجستيرون (29%) 11 ألفا ، 17 ألفا - ثنائي هيدروكسي البروجستيرون (2.5%

The paradox of pregnancy : an update on the immunology of early pregnancy

Pregnancy is an altered physiological state where an organism essentially foreign to the individual carrying it, grows, develops and at an appropriate time probably initiates a series of signals which lead to its safe expulsion from the woman's body. The immunological changes which allow this process are unique to pregnancy. Recent work in this field has led to a further understanding of the changes which operate to adapt the woman to the pregnant state. The concept that has developed over the years is one where a number of factors exert their effect both at the systemic but mostly at the local uterine level to modulate the immune response which will then refrain from mounting an inflammatory response against the invading trophoblast. The main protagonists of this immunomodulation are embryonic factors, uterine (endometrial) NK cells and, of course, the hormone progesterone. Progress has been made from the original observations of miscarriage rates in HLA sharing couples and with the possibility of research in couples undergoing IVF cycles, factors are being identified which initiate immunomodulation. Once implantation occurs the endometrial NK cells which are abundant from the late luteal phase are activated to control trophoblastic invasion and enhance the changes in blood vessels which allow for adequate feto-maternal perfusion. The immune response is controlled by PIBF under the influence of progesterone to bias towards a humoral response and suppress a cytotoxic response. All these processes are prone to fail at times and the clinical manifestation of such a failure is miscarriage along with other obstetric complications such as intra-uterine growth retardation, pre-eclampsia and placental abruption. Progress in the understanding of the immunological processes which protect pregnancy will help in elucidating the mechanisms whereby these processes fail. A consequence of this should be the explanation of those cases as yet classified as unexplained recurrent miscarriage. The literature indicates that the prognosis for this group of patients is not as encouraging as one would hope and that progress in this area is eagerly awaited by both patients and doctors working in this field.peer-reviewe

Peripheral and central mechanisms involved in hormonal control of male and female reproduction

Reproduction involves the integration of hormonal signals acting across multiple systems togenerate a synchronized physiological output. A critical component of reproduction is the luteinizinghormone (LH) surge, which is mediated by estradiol (E2) and neuroprogesterone interacting tostimulate kisspeptin release in the rostral periventricular nucleus of the third ventricle in rats. Recentevidence has shown that both classical and membrane E2 and progesterone signaling is involved inthis pathway. A metabolite of gonadotropin-releasing hormone (GnRH), GnRH-(1-5), has been shownto stimulate GnRH expression, secretion, and has a role in the regulation of lordosis. Additionally,gonadotropin-inhibitory hormone (GnIH) projects to and influences the activity of GnRH neurons inbirds. Stress-induced changes in GnIH have been shown to alter breeding behaviors in birds,demonstrating another molecular control of reproduction. Peripherally, paracrine and autocrineactions within the gonad have been suggested as therapeutic targets for infertility in both males andfemales. Dysfunction of testicular prostaglandin synthesis is a possible cause of idiopathic maleinfertility. Indeed, local production of melatonin and corticotropin-releasing hormone (CRH) couldinfluence spermatogenesis via immune pathways in the gonad. In females, vascular endothelialgrowth factor A (VEGF-A) has been implicated in an angiogenic process that mediates developmentof the corpus luteum and thus fertility via the Notch signaling pathway. Age-induced decreases infertility involve ovarian kisspeptin and its regulation of ovarian sympathetic innervation. Finally,morphological changes in the arcuate nucleus of the hypothalamus influence female sexualreceptivity in rats. The processes mediating these morphological changes have been shown toinvolve rapid effects of E2 controlling synaptogenesis in this hypothalamic nucleus. Together, thisreview highlights new research in these areas, focusing on recent findings in the molecularmechanisms of central and peripheral hormonal control of reproduction.Fil: Rudolph, L. M.. University of California at Los Angeles; Estados UnidosFil: Bentley, G. E.. University of California Berkeley; Estados UnidosFil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Paredes, A. H.. Universidad de Chile; ChileFil: Tesone, Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Wu, T. J.. Uniformed Services University; Estados UnidosFil: Micevych, P. E.. University of California at Los Angeles; Estados Unido