Gestational Trophoblastic Neoplasia Following Molar Ectopic Pregnancy A Case Report

BACKGROUND: Ectopic molar pregnancy is a gestational trophoblastic disease (GTD) of rare occurrence and therefore not always remembered as a diagnostic possibility.CASE: We describe a case of molar ectopic pregnancy in a primiparous woman who developed gestational trophoblastic neoplasia and required chemotherapy to achieve remission.CONCLUSION This case stresses the important role of histopathologic examination in establishing a diagnosis of ectopic molar pregnancy. Moreover, close follow-up of human chorionic gonadotropin levels is required when a GTD is suspected. (J Reprod Med 2008;53:579-582)

Características das Curvas de Regressão da Gonadotrofina Coriônica Pós-mola Hidatiforme Completa

Objetivos: construir a curva de regressão do b-hCG pós-mola hidatiforme completa (MHC) com remissão espontânea e comparar com a curva de regressão pós-MHC com tumor trofoblástico gestacional (TTG). Análise comparativa da curva de regressão do b-hCG das portadoras de MHC, acompanhadas no Serviço, com a curva de regressão observada por outros autores1-3. Métodos: foi realizada avaliação clínica e laboratorial (dosagem sérica de b-hCG), na admissão e no segmento pós-molar, de todas as pacientes com MHC, atendidas entre 1990 e 1998 no Hospital das Clínicas de Botucatu - Unesp. O resultado da determinação seriada do b-hCG foi analisado em curvas log de regressão. A evolução da curva de regressão do b-hCG foi analisada e comparada em MHC com remissão espontânea e MHC com TTG numa curva log de regressão, com intervalo de confiança de 95%. A curva log de regressão do grupo de remissão espontânea foi comparada com curvas consideradas padrão1,2. Foram construídas curvas log individuais de todas as pacientes e classificadas de acordo com os quatro tipos de curva (I, II, III e IV), propostos para o seguimento pós-molar³. Resultados: 61 pacientes com MHC tiveram seguimento pós-molar completo, 50 (82%) apresentaram remissão espontânea e 11 (18%) desenvolveram TTG. No grupo de pacientes com MHC e remissão espontânea, o tempo para alcançar a normalização dos níveis do b-hCG, após o esvaziamento molar, foi até 20 semanas. As pacientes que desenvolveram TTG apresentaram desvio precoce da curva de regressão normal do b-hCG, 4 a 6 semanas após o esvaziamento molar. Nestas pacientes, a quimioterapia foi introduzida em média na 9ª semana pós-esvaziamento molar. Conclusões: a curva de regressão do b-hCG pós-MHC com remissão espontânea apresentou declínio log exponencial, semelhante ao observado por outros autores1,2, e diferente das MHC com TTG. Foram identificados três tipos de curvas de regressão do b-hCG, semelhantes aos de Goldstein³, I, II e IV, e outros dois tipos diferentes de regressão do b-hCG: V (regressão normal) e VI (regressão anormal).

Preditores Clínicos e Histopatológicos de Tumor Trofoblástico Gestacional pós-Mola Hidatiforme Completa

Objetivo: definir os preditores clínicos e histopatológicos mais eficientes da evolução da mola hidatiforme completa (MHC) para tumor trofoblástico gestacional (TTG). Métodos: estudo prospectivo clínico e histopatológico de todas as portadoras de MHC, atendidas entre 1990 e 1998 no Hospital das Clínicas de Botucatu -- UNESP. A avaliação clínica pré-esvaziamento molar classificou a gravidez molar em: MHC de alto risco e MHC de baixo risco. Foram analisados os preditores clínicos para TTG, estabelecidos por Goldstein et al.¹ e por outros autores2--10. A avaliação histopatológica incluiu a determinação do diagnóstico de MHC, segundo os critérios de Szulman e Surti11, e o reconhecimento dos fatores de risco para TTG, de Ayhan et al.8. Os preditores clínicos e histopatológicos foram correlacionados com o desenvolvimento de TTG pós-molar. Resultados: em 65 portadoras de MHC, cistos do ovário maiores que 6 cm e tamanho uterino maior que 16 cm foram os preditores clínicos mais eficientes de TTG. A proliferação trofoblástica, a atipia nuclear, a necrose/hemorragia, a maturação trofoblástica e a relação cito/sinciciotrofoblasto não foram preditores significativos para TTG. A correlação entre preditor clínico e histopatológico para o desenvolvimento de TTG não foi possível porque nenhum parâmetro histopatológico foi significativo. Conclusões: mais estudos são necessários para avaliar possíveis preditores de persistência (TTG) e sua aplicação no contexto clínico das MHC. Enquanto isso, a determinação seriada de hCG sérico permanece o único indicador prognóstico seguro para TTG pós-MHC.

Strategies to improve fertility in Bos indicus postpubertal heifers and nonlactating cows submitted to fixed-time artificial insemination

Two experiments were designed to evaluate strategies to increase fertility of Bos indicus postpubertal heifers and nonlactating cows submitted to a fixed-time artificial insemination (TAI) protocol consisting of an intravaginal device containing 1.9 g of progesterone (CIDR) insertion + estradiol benzoate on Day 0, CIDR withdrawal + estradiol cypionate on Day 9, and TAI on Day 11. In Experiment 1, heifers (n = 1153) received a new or an 18-d previously used CIDR and, on Day 9, prostaglandin F(2 alpha) (PGF(2 alpha)) + 0, 200, or 300 IU equine chorionic gonadotropin (eCG). Heifers treated with a new CIDR had greater (least squares means +/- SEM) serum concentration of progesterone on Day 9 (3.06 +/- 0.09 ng/mL vs. 2.53 +/- 0.09 ng/mL; P < 0.05) and a smaller follicle at TAI (11.61 +/- 0.11 nim vs. 12.05 +/- 0.12 mm; P < 0.05). Heifers with smaller follicles at TAI had lesser serum progesterone, concentrations on Day 18 and reduced rates of ovulation, conception, and pregnancy (P < 0.05). Treatment with eCG improved (P < 0.05) follicle diameter at TAI (11.50 +/- 0.10 mm, 11.90 +/- 0.11 mm, and 12.00 +/- 0.10 mm, for 0, 100, and 200 IU, respectively), serum progesterone concentration on Day 18 (2.77 +/- 0.11 ng/mL, 3.81 +/- 0.11 ng/mL, and 4.87 +/- 0.11 ng/mL), and rates of ovulation (83.8%, 88.5%, and 94.3%) and pregnancy (41.3%, 47.0%, and 46.7%). In Experiment 2, nonlactating Nelore cows (n = 702) received PGF(2 alpha) treatment on Days 7 or 9 and, on Day 9, 0 or 300 IU cCG. Cows receiving PGF(2 alpha) on Day 7 had lesser serum progesterone concentrations on Day 9 (3.05 +/- 0.21 ng/mL vs. 4.58 +/- 0.21 ng/mL; P < 0.05), a larger follicle at TAI (11.54 +/- 0.21 mm vs. 10.84 +/- 0.21 mm; P < 0.05), and improved (P < 0.05) rates of ovulation (85.4% vs. 77.0%), conception (60.9% vs. 47.2%), and pregnancy (52.0% vs. 36.4%). Treatment with eCG improved (P < 0.05) serum progesterone concentration on Day 18 (3.24 +/- 0.14 ng/mL vs. 4.55 +/- 0.14 ng/mL) and the rates of ovulation (72.4% vs. 90.0%) and pregnancy (37.5% vs. 50.8%). In conclusion, giving PGF(2 alpha) earlier in the protocol in nonlactating cows and eCG treatment in postpubertal heifers and nonlactating cows improved fertility in response to a TAI (progesterone + estradiol) protocol. (C) 2009 Elsevier B.V. All rights reserved.

Progesterone-based estrous synchronization protocols in non-suckled and suckled primiparous Bos indicus beef cows

In Experiment 1, effects of an intravaginal progesterone insert (CIDR) for synchronization of estrus in non-suckled cycling primiparous Bos indicus beef cows were evaluated. Cows received either no treatment (Control), prostaglandin F(2 alpha) (PGF), or CIDR (6 d)+prostaglandin F(2 alpha) (CIDR+PGF). There were greater (P<0.05) percentages of cows inseminated and pregnant in 4 d of breeding season in CIDR+PGF (81.6%; 48.7%) than in PGF (53.8%; 32.5%) and Control cows (14.6%; 8.5%). In Experiment 2, efficacy of a treatment with CIDR (6 d)+prostaglandin F(2 alpha)+48 h temporary weaning (CIDR+PGF+TW) associated or not with equine chorionic gonadotropin (eCG) for synchronization of estrus in suckled primiparous B. indicus beef cows was evaluated. Cows were assigned to receive either no treatment (Control), CIDR+PGF+TW, CIDR+PGF+TW+200 IU of eCG, or CIDR+PGF+IW+400 IU of eCG. There were greater (P<0.05) percentages of insemination and pregnancy in 4 d of breeding season in treatment groups (47.0%; 27.5%) than in Control cows(4.5%; 3.0%) and in cows treated with eCG (50.9%; 29.4%) than in cows treated only with CIDR+PGF+TW (39.4%: 23.7%). No effects of eCG dosage were detected. The percentage of cows inseminated and pregnant in a 30 d breeding season were greater (P<0.05) in cows assigned to synchronization treatments (53.3%; 37.2%) than in Control cows (35.3%; 21.4%). In the beginning of the BS, non-suckled primiparous cows treated with CIDR+PGF with estrous cycles having been initiated following calving had greater estrous detection and pregnancy rates than PGF and Control cows, and suckled primiparous cows had improved estrous detection and pregnancy rates when eCG was associated with CIDR+PGF+TW. (C) 2009 Elsevier B.V. All rights reserved.

Fixed-time artificial insemination with estradiol and progesterone for Bos indicus cows II: Strategies and factors affecting fertility

In Experiments 1, 2, and 3, we evaluated the effects of temporary weaning (TW), equine chorionic gonadotropin (eCG), and follicle-stimulating hormone (FSH) treatments on results of it fixed-time artificial insemination (TAI) protocol in postpartum Bos indicus cows. In Experiment 1, treatment with 400 IU eCG or with TW for 48 h consistently improved pregnancy rates (PRs) at TAI, but, in Experiment 2, FSH treatment was less effective than eCG or TW. In Experiment 3, the inclusion of eCG treatment in cows subjected to TW did not improve PRs. We concluded that TW or 400 IU eCG should be included in the TAI protocol in postpartum Bos indicus cows to enhance fertility. In Experiment 4, we used records front heifers and cows treated with the proposed protocol during the 2006-2007 (n = 27,195) and 2007-2008 (n = 36,838) breeding seasons from multiple locations in Brazil to evaluate factors potentially affecting PRs. Overall PR at TAI was 49.6% (31,786 of 64,033). Pregnancy rate differed (P < 0.01) among farm within location (results ranging between 26.8% and 68.0%; P < 0.01), cow group within farm, by breed (Bos indicus, 48.3% [26,123 of 54,145]; Bos taurus, 61.7% [3652 of 5922]; and crossbred Bos indicus x Bos taurus, 50.7% [2011 of 3966]), category (nulliparous, 39.6% [2095 of 52901] suckled primiparous, 45.2% [3924 of 8677]; suckled multiparous, 51.8% [24,245 of 46,767]; and nonsuckled multiparous, 46.1% [1522 of 3299]), body condition score at TAI (<= 2.5 43.0% [3409 of 7923]; 3.0, 49.6% [18,958 of 38,229]; and >3.5, 52.7% [9419 of 17,881]). Days postpartum at beginning, of protocol did not affect PR (30 to 60 d, 47.6% [4228 of 8881]; 61 to 90 d, 51.7% 116,325 to 31,5721; and 91 to 150 d, 50.8% [7616 to 14,991]; P > 0.1). Pregnancy rate was also consistently affected (P < 0.01) by sire (results ranging from 7.2% to 77.3%) and artificial insemination technician (results ranging from 15.1% to 81.8%). (C) 2009 Elsevier B.V. All rights reserved.

Effects of postbreeding gonadotropin treatments on conception rates of lactating dairy cows subjected to timed artificial insemination or embryo transfer in a tropical environment

The objective of experiment 1 was to evaluate the effects of treatments with human chorionic gonadotropin (hCG) or GnRH 7 d after induced ovulation on reproductive performance of lactating dairy cows submitted to timed artificial insemination (TAI) or timed embryo transfer (TET). A total of 834 potential breedings were used from 661 lactating Holstein cows (37.3 +/- 0.3 kg of milk/d). Cows had ovulation synchronized and were assigned randomly to receive TAI on d 0 or TET on d 7. Within each group, cows were assigned randomly to receive on d 7 no additional treatment (control; n(TAI) = 156; n(TET) = 126), a 100 mu g i.m. injection of GnRH (n(TAI) = 155; n(TET) = 124), or a 2,500 TU i.m. injection of hCG (ITA = 151; n(TET) = 122). Postbreeding treatment affected the percentages of pregnant cows at TET on d 28 (control: 38.1%; GnRH: 52.4%; hCG: 45.1%) and on d 60 (control: 32.5%; GnRH: 41.1%; hCG: 38.5%), but postbreeding treatment did not affect percentages of pregnant cows at TAT on d 28 (control: 30.1%; GnRH: 32.2%; hCG: 32.4%) or on d 60 (control: 25.6%; GnRH: 27.1%; hCG: 29.8%). The objective of experiment 2 was to evaluate the effect of a treatment with GnRH 7 d after TET on reproductive performance of lactating dairy cows that received a previous GnRH treatment at TET. A total of 285 potential breedings were used from 257 lactating Holstein cows (35.1 +/- 0.8 kg of milk/d). Cows had ovulation synchronized and were assigned for TET on d 7. Immediately after TET, all cows were treated with a 100 mu g i.m. injection of GnRH. on d 14, cows were assigned randomly to receive (G7-14; n = 147) or not (G7; n = 138) an additional injection of GnRH. Pregnancy diagnosis were performed on d 28 and 60. The additional treatment with GnRH on d 14 did not affect the percentages of pregnant cows on d 28 (G7: 48.5%; G7-14: 42.9%) or on d 60 (G7: 39.8%; G7-14: 37.4%). In conclusion, treatment with GnRH or hCG 7 d after induced ovulation increased conception rates in lactating dairy cows submitted to TET, but not in cows submitted to TAI. Moreover, treatment with GnRH 7 d after TET did not enhance reproductive performance of lactating dairy cows that received a previous GnRH treatment at TET.

Use of oocyte transfer in a commercial breeding program for mares with reproductive abnormalities

In some mares with lesions of the reproductive tract, embryo collection and survival rates are low or collection of embryos is not feasible. For these mares, oocyte transfer has been proposed as a method to induce pregnancies. In this report, a method for oocyte transfer in mares and results of oocyte transfer performed over 2 breeding seasons, using mares with long histories of subfertility and various reproductive lesions, are described.Human chorionic gonadotropin or an implant containing a gonadotropin-releasing hormone analog was used to initiate follicular and oocyte maturation. Oocytes were collected by means of transvaginal ultrasound-guided follicular aspiration. Following follicular aspiration, cumulus oocyte complexes were evaluated for cumulus expansion and signs of atresia; immature oocytes were cultured in vitro to allow maturation. The recipient's ovary and uterine tube (oviduct) were exposed through a flank laparotomy with the horse standing, and the oocyte was slowly deposited within the oviduct. Oocyte transfer was attempted in 38 mares between 9 and 30 years old during 2 successive breeding seasons. All mares had a history of reproductive failure while in breeding and embryo transfer programs. Twenty pregnancies were induced. Fourteen of the pregnant mares delivered live foals. Results suggest that oocyte transfer can be a successful method for inducing pregnancy in subfertile mares in a commercial setting..

The efficacy of a single chain recombinant equine luteinizing hormone (reLH) in mares: Induction of ovulation, hormone profiles, and inter-ovulatory intervals

The objectives of this study were to determine the efficacy of recombinant equine luteinizing hormone (reLH) in shortening the time to ovulation in cycling mares and to determine the effects of treatment on endogenous hormones and inter-ovulatory intervals. In study 1, mares of light horse breeds (3-20 years) were treated with either a vehicle, various doses of reLH, or human chorionic gonadotropin (hCG). Cycling mares were examined by palpation and ultrasound per rectum daily or every 12 h from the time of treatment to ovulation. In studies 2 and 3, jugular blood samples were collected daily or every 12 h from the time of treatment to ovulation for analysis of LH, follicle stimulating hormone (FSH), estradiol-17 beta (E-2), and progesterone (P-4) by radioimmunoassays (RIA). Increasing doses of reLH (0.3, 0.6, 0.75, and 0.9 mg) showed increasing effectiveness at inducing ovulation within 48 h of treatment. Treatments with the 0.75 and 0.9 mg doses of reLH resulted in 90% and 80% ovulation rates, which were similar to hCG treatment (85.7%). Except for the early rise in LH after treatment with 0.5, 0.65, and 1.0 mg of reLH, hormone profiles appeared to be similar between control and treated cycles. Inter-ovulatory intervals were similar between control and treatment cycles. In conclusion, reLH is a reliable and effective ovulatory agent that does not significantly alter endogenous hormone profiles or affect inter-ovulatory intervals.(c) 2007 Elsevier B.V. All rights reserved.

Effect of eFSH on ovarian cyclicity and embryo production of mares in spring transitional phase

The use of equine FSH (eFSH) for inducing follicular development and ovulation in transitional mares was evaluated. Twenty-seven mares, from 3 to 15 years of age, were examined during the months of August and September 2004, in Brazil. Ultrasound evaluations were performed during 2 weeks before the start of the experiment to confirm transitional characteristics (no follicles larger than 25 mm and no corpus luteum [CL] present). After this period, as the mares obtained a follicle of at least 25 mm, they were assigned to one of two groups: (1) control group, untreated; (2) treated with 12.5 mg eFSH, 2 times per day, until at least half of all follicles larger than 30 mm had reached 35 mm. Follicular activity of all mares was monitored. When most of the follicles from treated mares and a single follicle from control mares acquired a preovulatory size ( : 35 mm), 2,500 IU human chorionic gonadotropin (hCG) was administered IV to induce ovulation. After hCG administration, the mares were inseminated with fresh semen every other day until ovulation. Ultrasound examinations continued until detection of the last ovulation, and embryo recovery was performed 7 to 8 days after ovulation. The mares of the treated group reached the first preovulatoiy follicle (4.1 +/- 1.0 vs 14.9 +/- 10.8 days) and ovulated before untreated mares (6.6 +/- 1.2 vs 18.0 +/- 11.1 days; P <.05). All mares were treated with prostaglandin F-2 alpha (PGF(2 alpha)), on the day of embryo flushing. Three superovulated mares did not cycle immediately after PGF(2 alpha), treatment, and consequently had a longer interovulatory interval (22.4 vs 10.9 days, P < 0.05). The mean period of treatment was 4.79 1.07 days and 85.71% of mares had multiple ovulations. The number of ovulations (5.6 vs 1.0) and embryos (2.0 vs 0.7) per mare were higher (P < 0.05) for treated mares than control mares. In conclusion, treatment with eFSH was effective in hastening the onset of the breeding season, inducing multiple ovulations, and increasing embryo production in transitional mares. This is the first report showing the use of FSH treatment to recover embryos from the first cycle of the year.

Use of corticosteroid therapy on the modulation of uterine inflammatory response in mares after artificial insemination with frozen semen

In order to modulate uterine inflammatory response and evaluate the effect of corticosteroid therapy on fertility, 90 cycles of 45 mares were used for artificial insemination with frozen semen, using three different protocols: G1 - inseminated with frozen semen (800 x 10(6) viable spermatozoa pre-freezing) + 20 mL of seminal plasma; G2 - inseminated with frozen semen (800 x 10(6) viable spermatozoa pre-freezing) + corticosteroid therapy; G3 - inseminated with frozen semen (800 x 10(6) viable spermatozoa pre-freezing) + 20 mL of seminal plasma + corticosteroid therapy. Corticosteroid therapy consisted on one administration of prednisolone acetate (0.1 mg/Kg - Predef (R)) when mares presented 35mm follicles and uterine edema, concomitantly with the unique dose of hCG (human chorionic gonadotropin), then repeated each 12 hours until ovulation. on first fertility trial, with normal mares, there was no difference between control and treated groups (p>0.05), using seminal plasma associated with corticosteroid therapy (40 vs. 38%, respectively) or corticosteroid therapy alone (40 vs. 45% respectively). The second fertility trial, performed with mares with previous history of post-insemination endometritis, demonstrated a significant increase of pregnancy rate when mares were submitted to corticosteroid therapy (0.0 vs. 64.5%, respectively; p<0.05). Corticosteroid therapy was shown to be safe, with no physical or reproductive alterations on treated mares, demonstrating to be an adequate option to those animals with history of post-breeding or post-insemination endometritis. Further clinical research is necessary to confirm these results and contribute to the establishment of preventive therapy for cases of post-insemination endometritis.

Use of a Low Dose of Equine Purified FSH to Induce Multiple Ovulations in Mares

The effects of a low dose of equine purified FSH (eFSH) on incidence of multiple ovulations and embryo recovery rate in mares were studied. During the physiological breeding season in Brazil (19 degrees 45'45'S), 14 Mangalarga Marchador donor mares were used in a crossover study and another 25 mares of the same breed, between 3 years and 12 years of age were used as recipients for the embryo transfers. Donors were monitored during two consecutive oestrus cycles, an untreated control cycle followed by a treated cycle, when eFSH was administered. In both cycles, after an embryo collection attempt on day 8 post-ovulation all mares received 7.5 mg dinoprost and had their two largest follicles tracked daily by ultrasonography until the period of ovulation. Mares were inseminated every 48 h with extended fresh semen from a single stallion after the identification of a 35-mm follicle until the period of ovulation. Ovulations were induced by intravenous administration of 2.500 IU of human chorionic gonadotropin, upon detection of a 35- to 40-mm follicle. In the treated cycle, 5 mg eFSH was given intramuscularly once a day, from day 8 post previous ovulation until at least one follicle reached 35 mm in diameter. Embryo flushes were performed on day 8 of dioestrus (day 0 = ovulation). Treatment with eFSH resulted in higher (p < 0.05) ovulation rate and incidence of multiple ovulations compared to the control (1.6 vs 1.0 and 50% vs 0%, respectively - one mare had triple ovulation). However, embryo recovery rates in the control and treated cycles were similar (0.8 and 1.0, respectively; p > 0.05). Pregnancy rates in the recipient mares following embryo transfer were similar for the control and eFSH cycles (11/11 and 10/14, respectively). Additional studies are necessary in order to develop a low-dose protocol for the use of eFSH that brings a more consistent contribution to the efficiency of commercial equine embryo transfer programs.

Effect of melatonin on DNA damage of bovine cumulus cells during in vitro maturation (IVM) and on in vitro embryo development

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Superovulation and embryo transfer in Bos indices cattle

Compared to Bos taurus breeds, Bos indices breeds of cattle present several differences in reproductive physiology. Follicular diameter at deviation and at the time of ovulatory capability are smaller in B. indicus breeds. Furthermore, B. indicus breeds have a greater sensitivity to gonadotropins, a shorter duration of estrus, and more often express estrus during the night. These differences must be considered when setting up embryo transfer programs for B. indicus cattle. In recent studies, we evaluated follicular dynamics and superovulatory responses in B. indicus donors with the objective of implementing fixed-time AI protocols in superstimulated donors. Protocols using estradiol and progesterone/progestrogen releasing devices to control follicular wave emergence were as efficacious as in B. taurus cattle, allowing the initiation of superstimulatory treatments (with lower dosages of FSH than in B. taurus donors) at a self-appointed time. Furthermore, results presented herein indicate that delaying the removal of progesterone/progestogen-releasing devices, combined with the administration of GnRH or pLH 12 h after the last FSH injection, results in synchronous ovulations, permitting the application of fixed-time AI of donors without the necessity of estrus detection and without compromising the results. (c) 2005 Elsevier B.V. All rights reserved.

Expression of protease nexin-1 and plasminogen activators during follicular growth and the periovulatory period in cattle

Extracellular matrix remodeling occurs during ovarian follicular development, mediated by plasminogen activators (PAs) and PA inhibitors including protease nexin-1 (PN-1). In the present study we measured expression/activity of the PA system in bovine follicles at different stages of development by timed collection of ovaries during the first follicular wave and during the periovulatory period, and in follicles collected from an abattoir. The abundance of mRNA encoding PN-1, tissue-type PA (tPA), urokinase (uPA) and PA inhibitor-1 (PAI-1) were initially upregulated by human chorionic gonadotropin (hCG) in bovine preovulatory follicular wall homogenates. PN-1, PAI-1 and tPA mRNA expression then decreased near the expected time of ovulation, whereas uPA mRNA levels remained high. PN-1 concentration in follicular fluid (FF) decreased and reached the lowest level at the time of ovulation, whereas plasmin activity in FF increased significantly after hCG. Follicles collected from the abattoir were classified as non-atretic, early-atretic or atretic based on FF estradiol and progesterone content: PN-1 protein levels in FF were significantly higher in non-atretic than in atretic follicles, and plasmin activity was correspondingly higher in the atretic follicles. No changes in PN-1 levels in FF were observed during the growth of pre-deviation follicles early in a follicular wave. These results indicate that PN-1 may be involved in the process of atresia in non-ovulatory dominant follicles and the prevention of precocious proteolysis in periovulatory follicles.