Bibliography: leaves 182-210.

<p>Extracellular matrix degrading proteases from the plasminogen (plg) activator (PA) and the matrix metalloproteinase (MMP) systems have been implicated as important mediators of ovulation and corpus luteum (CL) formation and regression. The aim of this thesis was to investigate the expression and regulation of PAs and MMPs in the ovary and to examine their functional roles for CL formation and function. </p><p> The expression of membrane-type MMP-1 (MT1-MMP) and its substrate gelatinase A (MMP-2) mRNAs was studied during pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-induced ovulation in immature rats. These proteases were coordinately regulated so that both were highly expressed in the theca cells of large preovulatory follicles. This suggests that MT1-MMP activates gelatinase A in preovulatory follicles to degrade the follicular wall during ovulation. </p><p> In pseudopregnant (psp) rats, MT1-MMP mRNA was expressed in the CL throughout the luteal phase. Tissue inhibitor of metalloproteases type-1 (TIMP-1) mRNA was expressed during CL formation and regression. MMP-2 and collagenase-3 mRNAs were expressed during CL formation and regression, respectively. When the luteal phase was artificially prolonged or shortened, TIMP-1 and collagenase-3 mRNAs were induced only after the serum progesterone levels had decreased, indicating a close association with luteolysis in the rat. </p><p> In psp mice, the expression of mRNAs coding for both PAs, seven MMPs, and five protease inhibitors was studied. Most of the studied molecules were coordinately expressed during formation or regression of the CL. However, uPA, MT1-MMP, and TIMP-3 mRNAs were expressed throughout the luteal phase. The role of uPA was examined in psp uPA deficient mice. These mice displayed no abnormalities in luteal function or vascularity. The role of uPA is thus either not essential or can be compensated by other proteases in the absence of uPA. </p><p> In order to control the timing of the CL formation, a mouse model for PMSG/hCG-induced CL formation was developed. Five different protocols were evaluated. One of them provided CL that were stable for six days. In that protocol the mice were treated with prolactin (PRL), twice daily from day 2 of CL life onward. The expression of the steroid acute regulatory protein (StAR) mRNA in the psp CL was also characterized to assess its use as a molecular marker for CL development and regression. It was highly expressed in the forming and functional CL and downregulated at a late stage of CL regression.</p><p> The functional role of plg and MMPs for CL formation and function was investigated in plg deficient mice treated with the MMP inhibitor galardin (GM6001). Both psp mice and PMSG/hCG +PRL-induced CL formation were used. Several molecular markers for CL development and regression were used to evaluate the health status of the CL. Our data showed that healthy and vascularized CL formed even in plg deficient mice treated with the inhibitor. However, serum progesterone levels were significantly reduced in these mice, an effect that was mainly attributable to the plg deficiency. In conclusion, neither plg nor MMPs, alone or in combination, seem to be essential for the development of a functional CL.</p>

Extracellular matrix degrading proteases from the plasminogen (plg) activator (PA) and the matrix metalloproteinase (MMP) systems have been implicated as important mediators of ovulation and corpus luteum (CL) formation and regression. The aim of this thesis was to investigate the expression and regulation of PAs and MMPs in the ovary and to examine their functional roles for CL formation and function. The expression of membrane-type MMP-1 (MT1-MMP) and its substrate gelatinase A (MMP-2) mRNAs was studied during pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-induced ovulation in immature rats. These proteases were coordinately regulated so that both were highly expressed in the theca cells of large preovulatory follicles. This suggests that MT1-MMP activates gelatinase A in preovulatory follicles to degrade the follicular wall during ovulation. In pseudopregnant (psp) rats, MT1-MMP mRNA was expressed in the CL throughout the luteal phase. Tissue inhibitor of metalloproteases type-1 (TIMP-1) mRNA was expressed during CL formation and regression. MMP-2 and collagenase-3 mRNAs were expressed during CL formation and regression, respectively. When the luteal phase was artificially prolonged or shortened, TIMP-1 and collagenase-3 mRNAs were induced only after the serum progesterone levels had decreased, indicating a close association with luteolysis in the rat. In psp mice, the expression of mRNAs coding for both PAs, seven MMPs, and five protease inhibitors was studied. Most of the studied molecules were coordinately expressed during formation or regression of the CL. However, uPA, MT1-MMP, and TIMP-3 mRNAs were expressed throughout the luteal phase. The role of uPA was examined in psp uPA deficient mice. These mice displayed no abnormalities in luteal function or vascularity. The role of uPA is thus either not essential or can be compensated by other proteases in the absence of uPA. In order to control the timing of the CL formation, a mouse model for PMSG/hCG-induced CL formation was developed. Five different protocols were evaluated. One of them provided CL that were stable for six days. In that protocol the mice were treated with prolactin (PRL), twice daily from day 2 of CL life onward. The expression of the steroid acute regulatory protein (StAR) mRNA in the psp CL was also characterized to assess its use as a molecular marker for CL development and regression. It was highly expressed in the forming and functional CL and downregulated at a late stage of CL regression. The functional role of plg and MMPs for CL formation and function was investigated in plg deficient mice treated with the MMP inhibitor galardin (GM6001). Both psp mice and PMSG/hCG +PRL-induced CL formation were used. Several molecular markers for CL development and regression were used to evaluate the health status of the CL. Our data showed that healthy and vascularized CL formed even in plg deficient mice treated with the inhibitor. However, serum progesterone levels were significantly reduced in these mice, an effect that was mainly attributable to the plg deficiency. In conclusion, neither plg nor MMPs, alone or in combination, seem to be essential for the development of a functional CL.

This dissertation project investigated fluctuations in estradiol and testosterone across the human menstrual cycle. During the part of the cycle when women are most fertile, women show stronger preferences for men with more masculine faces, and these preference changes may be related to changes in hormone levels during ovulation. The present study investigated preferences changes among women for higher testosterone men over the menstrual cycle as estradiol and testosterone in those women fluctuated. 32 women participated in this 5-week long study tracking their estradiol and testosterone levels and preferences for masculine men. Women with higher levels of estradiol preferred men who had higher levels of testosterone than women who had lower levels of estradiol. During ovulation, women were more like to find high testosterone men more attractive than other parts of the menstrual cycle. In addition to ratings of men, several other psychological tests were administered and examined for changes as a function of state and trait levels of hormones.

Šio darbo tikslas buvo nustatyti ovuliacijos pasireiškimo laiko priklausomybę nuo kalės veislės dydžio ir amžiaus bei metų laiko, bei palyginti GnRH agonisto Suprelorin implanto sukeltos ovuliacijos pasireiškimo laiką su natūraliai pasireiškusios rujos ovuliacijos laiku. Šiam tikslui pasiekti pirmiausiai buvo ištirta 194 kalių (51 skirtingos veislės) progesterono koncentracija kraujo serume. Iš šių kalių buvo atrinktos tos kalės, kurioms pagal progesterono koncentraciją pasireiškė ovuliacija (53 kalės). Nustatyta, kad progesterono koncentracija kraujo serume siekė 20-30 nmol/l. Kalės buvo suskirstytos į grupes: pagal amžių (2 grupės – iki 5 metų (n=41); 5 metų ir vyresnės (n=12) ), pagal veislės dydį (3 grupės – mažos (n=22), vidutinės (n=14) ir didelės (n=17) veislės), pagal sezoną (4 grupės – žiema (n=3), pavasaris (n=16), ruduo (n=11), vasara (n=23) ). Pagal kiekvieną požymį statistiškai buvo įvertinti ovuliacijos pasireiškimo laiko skirtumai tarp grupių, bei įvertinta ovuliacijos priklausomybė nuo kalės amžiaus, dydžio, metų laiko. Statistiškai patikimų skirtumų tarp grupių ir koreliacinių ryšių tarp ovuliacijos pasireiškimo laiko ir kalės amžiaus ir dydžio, bei metų laiko nebuvo nustatyta. Panašių tyrimų iki šiol buvo atlikta labai mažai, todėl informacija yra vertinga. Paskutiniame tyrimųetape4 kalėms buvo panaudoti GnRH agonisto Suprelorin implatai rujai sukelti. Implantas buvo įvedamas po oda, bambos srityje. Po įvedimo buvo imamas kraujas kelis kartus kas 3-5 dienas... [toliau žr. visą tekstą] / The main goal of this paper was to determine whether or not there is a significant influence of GnRH agonist, seasonality, age and size of the bitch on ovulation time, and to compare ovulation time induced by GnRH agonist Suprelorin implant and the ovulation time that occured naturally. To achieve this goal we tested the blood of 194 bitches (51 breeds) for progesterone levels in blood serum. Out of these 194 bitches we selected the ones (53 bitches) that ovulated. Meaning, the progesterone levels for these 53 bitches varied between 20-30 nmol/l. These 53 bitches were divided into groups: by age (2 groups – younger than 5 years (n=41); 5 years old and older (n=12),, by size (3 groups – small (n=22), medium (n=14), large (n=17), and by season (4 groups – winter (n=3), spring (n=16), summer (n=11), autumn (n=23). All the groups were statistically analyzed to determine whether or not there is a significant difference between the groups, and to determine if there are correlations between the ovulation time and the size and age of the bitch and time of year (season). No significant difference or correlations were determined. We have found a little information about influence of seasonality, and age and size of the bitch on ovulation time, so the information in this paper is valuable. For the last step to achieving the goal of the paper, GnRH agonist Suprelorin implant was used on 4 bitches. The agonist was implanted subcutaneously around the navel area. After the implantation... [to full text]

In der vorliegenden Arbeit sollte die Wirksamkeit von Altrenogest (Regumate®) und Gonadotropinen bzw. des GnRH-Analogons D-Phe6-GnRH im Rahmen eines Verfahrens zur Brunst- und Ovulationssynchronisation bei besamten, als ingravid detektierten Jung- und Altsauen überprüft werden. Die Untersuchungen wurden an insgesamt 265 Jung- und 542 Altsauen vorgenommen. Die Sauen wurden im Rahmen der ultrasonografischen Trächtigkeitskontrolle, die zwischen den Tagen 21 und 35 nach der ersten künstlichen Besamung durchgeführt wurde, als ingravid detektiert und einer von drei verschiedenen Versuchsgruppen (VG) zugeteilt: VG 1 (n = 490): Applikation von 4 ml Regumate®/Tier/Tag oral über 15 Tage; einmalige subkutane (s.c.) Injektion von 1.000 IE PMSG/eCG 24 Stunden nach letztmaliger Regumate®-Gabe; i.m. Applikation von 500 IE hCG 78 bis 80 h nach PMSG/eCG zur Induktion der Ovulation; VG 2 (n = 135): identisch zu VG 1, aber Gabe von 5 ml Regumate® und 800 IE PMSG; VG 3 (n = 182): identisch zu VG 2, aber Injektion von 50 µg D-Phe6-GnRH zur Induktion der Ovulation. Die Sauen wurden jeweils zweimal im Abstand von 24 und 40 Stunden nach hCG bzw. D-Phe6-GnRH künstlich besamt. Der Erfolg der Behandlungen wurde anhand der sonografischen Untersuchung der Ovarien kontrolliert. Untersuchungen erfolgten zum Zeitpunkt der Trächtigkeitsuntersuchung, am Ende der Regumate®-Behandlung, unmittelbar vor der ersten und unmittelbar nach der zweiten künstlichen Besamung. Trächtigkeits- (TR) und Abferkelrate (AFR) sowie Anzahl insgesamt und lebend geborener Ferkel wurden dokumentiert. Als jeweilige Kontrolltiere dienten Jung- bzw. Altsauen, die brunst- und ovulationssynchronisiert wurden, zur Erstbesamung anstanden und zeitgleich wie die Tiere der Versuchsgruppe besamt wurden. Die Auffindungsrate der Ovarien betrug 88,9 % zum Zeitpunkt der Trächtigkeitsuntersu-chung (TU) und 98,3 % am Ende der Regumate®-Behandlung. Während der Besamungen konnten bei allen untersuchten Tieren die Ovarien dargestellt werden. Zum Zeitpunkt der TU wiesen die Sauen überwiegend Corpora lutea (CL; 56,3 %; p < 0,05) auf. Der zweithäufigste Befund waren Follikel von 2-6 mm Durchmesser (F2-6; 27,7 %; p < 0,05). Durch Altrenogest konnte das Follikelwachstum effektiv gehemmt werden, ohne die spontane Luteolyse zu beeinträchtigen. Mehr als 84 % der Tiere wiesen am Ende der 15-tägigen Behandlung mit Regumate® F2-6 auf. Altsauen, die 4 ml Regumate® erhielten, hatten häu-figer als die mit 5 ml behandelten Tiere polyzystisch degenerierte Ovarien (POD). Zahlrei-che Altsauen, vor allem solche mit periovulatorischen Funktionsgebilden zum Zeitpunkt der Trächtigkeitsuntersuchung (p < 0,05), wiesen zudem CL am Ende der Behandlung auf (16,3 %). Jungsauen ovulierten unabhängig von der ovulationsauslösenden Substanz überwiegend zwischen beiden Besamungen (p < 0,05), während Altsauen zu gleichen Anteilen zwischen den Besamungen und nach der zweiten Besamung ovulierten. Die ovulationsauslösende Substanz, d.h. hCG bzw. GnRH, hatte weder bei Jung- noch bei Altsauen einen Einfluss auf das Ovulationsverhalten. Der Ovulationszeitpunkt war ohne Einfluss auf die Trächtig-keitsrate. Altsauen, die D-Phe6-GnRH erhielten, wurden häufiger tragend als Tiere, die hCG erhielten (p < 0,05). Reziprokes Ergebnis erzielten die Jungsauen (p < 0,05). Kon-trolltiere wiesen Trächtigkeits- bzw. Abferkelraten auf, die bis zu 15,6 % (TR; p < 0,05) bzw. 16,8 % (AFR) höher als die der Versuchstiere waren, da vor allem die Altsauen, nicht aber Jungsauen aller drei Versuchsgruppen schlechtere Fruchtbarkeitsleistungen erzielten. Aus den Ergebnissen ist zu schlussfolgern, dass die kombinierte Anwendung von Altrenogest (Regumate®) und Gonadotropinen bzw. GnRH-Analogon D-Phe6-GnRH zur Brunst- und Ovulationssynchronisation bei besamten, als ingravid detektierten Jung- und Altsauen geeignet ist. Ausgehend von den Ergebnissen ist zu empfehlen, Sauen mit 5 ml Reguma-te®/Tier/Tag zu behandeln (Zyklusblockade). Ein Applikationsintervall von 15 Tagen ist ausreichend. Werden 800 IE PMSG/eCG 24 Stunden nach letztmaliger Regumate®-Applikation verabreicht, kann mit zuverlässiger Stimulation des Follikelwachstums gerechnet werden. Jungsauen sollten 500 IE hCG 78 bis 80 Stunden nach der PMSG/eCG-Gabe erhalten. Bei Altsauen sind dazu 50 µg des GnRH-Analogons D-Phe6-GnRH emp-fehlenswert. Die transkutane Ultrasonografie ist ein geeignetes Verfahren zur Darstellung von Ovarien und ovarieller Funktionskörper.

Bibliography: leaves 182-210 / vii, 210 leaves : ill ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--Dept. of Obstetrics and Gynaecology, University of Adelaide, 1985

Master of Medicine / The number of unwanted pregnancies has not decreased in recent years and this should be addressed. Emergency contraception may be effective when used correctly having the advantage that it can be used after an episode of unprotected sexual intercourse (when regular contraception has failed or was not used). In this research project I set out to explore some of the major reasons why there are still many unwanted pregnancies in Australia. I decided to focus on the use and non-use of emergency contraception, e.g. emergency contraception pill (ECP) “method failures” are not well understood because the actual mechanisms of action are still unclear. There is evidence ECP can effectively interfere with follicle growth and ovulation. It is much less clear is whether ECP is able to interfere with fertilization and implantation, in a way, which may make it acceptable to those who have strong religious beliefs in fertilization being the start of new life. Emergency contraception has the potential to prevent many unwanted pregnancies when unprotected intercourse has occurred. It has relatively high efficacy in many studies, but true method failures are not well understood. By contrast, many unwanted pregnancies occur for “social reasons” where emergency contraception has not been used. I set out to study changes in knowledge and usage of emergency contraception in these groups of Australian women seeking termination of pregnancy: 1. Before a dedicated emergency contraception pill (ECP) pack (Postinor) became available in Australia 2. One year after dedicated ECP became available on prescription 3. One year after the ECP pack became available “over the counter” without prescription. Ninety-nine women were recruited during their presentation with a request for ECP at the six Family Planning Clinics in Australia. All women took LNG 1.5mg in a single dose during the clinic consultation. A blood sample was taken immediately prior to ingestion of the ECP for estimation of serum LH, oestradiol and progesterone levels to calculate the day of the menstrual cycle. Based on these endocrine data we estimated the timing of ovulation to within a ±24-hour period with an accuracy of around 80%. Women were followed up 4-6 weeks later to ascertain pregnancy status. The effectiveness of ECP when taken before and after ovulation was determined. Three women in this study became pregnant despite taking the ECP (pregnancy rate 3%). All three women who became pregnant had unprotected intercourse between day -1 and 0 and took the ECP on day +2, based on endocrine data. Day zero was taken as ovulation day. Among seventeen women who had intercourse in the fertile period of the cycle and took the ECP after ovulation occurred (on day +1 to +2) we could have expected 3 or 4 pregnancies, based on Wilcox et al data. Three pregnancies were observed. Among 34 women who had intercourse on days –5 to –2 of the fertile period, and took ECP before or around ovulation, four pregnancies could have been expected, but none were observed. The major discrepancies between women’s self-report of stage of the cycle and the dating calculation based on endocrine data were observed in this study. These data are supportive of the concept that the LNG ECP has little or no effect on post-ovulation events, but is highly effective before ovulation. Our interpretation of the data in terms of timing of treatment relative to ovulation may explain why EC with LNG works sometimes and fails at other times. A larger study is needed to prove this hypothesis. To investigate other reasons for such a high rate of unwanted pregnancy, which probably has a larger impact we looked into womens knowledge of and attitude towards ECP. Seven hundred and eighteen women participated in this study by answering a questionnaire consisting of 15 questions on their demographic and reproductive characteristics as well as the knowledge about the ECP, e.g. 208 women were enrolled before the ECP was marketed in Australia in 2001, 308 after it was marketed and 202 after it became available over the counter (Group 1, 2, and 3, respectively). We found that the participants who have heard about ECP were significantly younger (p<0.005). The mean age of women who have never heard about of ECP was 29.8 years compared to 26.3 years in women who have heard about ECP. More women were aware about the ECP after it became available over the counter. Women in group 2 had higher educational level in comparison to women in group 2 and 3 (p<0.005). There was significant trend in increased use of ECP in women of higher educational level (p<0.005). The use of ECP did not increase significantly with improved availability and access to the ECP amongst women presenting for termination of pregnancy. Wider availability of he ECP pack in Australia and an easier access to it has increased women’s awareness about the ECP. However, the use of ECP has not increased. This study provides better understanding of mechanism of action of LNG ECP and an explanation to the method failure. It also reveals poor knowledge about ECP despite its wider availability and accessibility. Improving these is a worldwide challenge for family planners and all health professionals.

Thesis (Ph. D.)--University of Virginia, 2008. / Includes bibliographical references. Also available in electronic form as viewed 2/16/2009.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2006. / Härtill 5 uppsatser.