Hormonas esteroideas foliculares y su relación con los resultados globales del ciclo de estimulación ovárica / Follicular steroid hormones and their relationship to the overall results of ovarian stimulation cycle

Objetivo: Relacionar los niveles de hormonas esteroideas foliculares con el ciclo de estimulación ovárica y sus resultados globales. Métodos: Se incluyeron pacientes < 38 años, con esterilidad de causa masculina, tubárica o desconocida, que recibieron un protocolo largo con agonistas de GnRH y rFSH. Se recogieron las muestras de la primera y segunda aspiración folicular de cada ovario y se realizó un quimioinmunoanálisis de estradiol, progesterona, testosterona y DHEAS. Resultados: Se obtuvieron cifras menores de DHEAS folicular en las pacientes con más días de frenado con agonistas de GnRH (p=0,0003). Cuantos más días de rFSH administrados, mayores fueron los niveles de testosterona y DHEAS folicular (p=0,03; p=0,03). En los resultados globales del ciclo, se obtuvo una correlación negativa entre las cifras de testosterona folicular y el número de complejos puncionados (r= -0,360; p=0,002) y entre la testosterona folicular y el número de embriones de calidad D (r= -0,233; p=0,047). El número de ovocitos maduros fue menor en pacientes con mayores niveles de testosterona folicular (p=0,008). La progesterona folicular fue superior en ovocitos de buena calidad frente a los de calidad no destacable (p=0,006) y muy mala calidad (p=0,04). Conclusiones: Las cifras altas de testosterona folicular se correlacionaron con menor número de complejos puncionados, ovocitos maduros y embriones de calidad D. La buena calidad ovocitaria se asoció a niveles de progesterona folicular superiores.

Objective: To relate the levels of follicular steroid hormones with the ovarian stimulation cycle and its overall results. Method: It was included patients < 38 years old with sterility of male, tubaric or unknown origin who underwent a long protocol with GnRH agonists and rFSH. Samples were obtained from the first and second follicular aspiration of each ovary. A chemiluminescent immunoassay of estradiol, progesterone, testosterone and DHEAS was performed. Results: Figures of follicular DHEAS decreased as the days of treatment with GnRH agonists increased (p=0.0003) and levels of follicular testosterone and DHEAS increased along with the days of treatment with rFSH (p=0.03, p=0.03). In regard to the outcomes of the overall cycle it was found a negative correlation between follicular testosterone levels and the number of punctured complexes (r= -0.360; p=0.002) and between follicular testosterone and the number of D quality embryos (r= -0.233; p=0.047). The number of mature oocytes was lower in patients with higher levels of follicular testosterone (p=0.008). Follicular progesterone was higher in good quality oocytes as compared to those of no remarkable quality (p=0.006) and very poor quality (p=0.04). Conclusions: High levels of follicular testosterone were correlated with a fewer number of punctured complexes, mature oocytes and D quality embryos. Good oocyte quality was associated with higher follicular progesterone levels.

relationship between follicular fluid androgen concentrations and its receptor expression in granulosa cells from human follicles

Polycystic ovary syndrome [PCOS] is the most common cause of oligoanovulation, infertility, and hyperandrogenism in women and characterized by abnormal folliculogenesis. The androgen receptoe [AR] is present in the ovary in almost all stages of folliculogenesis and has been suggested to play a proliferative role for follicular development but the role of androgen signaling through the AR in the pathophysiology of PCOS is still unclear. The aim of this study was to explore the role of androgens during folliculogenesis by determining AR mRNA expression in granulosa cell [GC] of human antral follicles in PCOS patients and controls and correlate that with the hormonal characteristics of the corresponding follicular fluid. The current study included 40 patients with PCOS and 30 women with normal ovulatory function attending the Fertility Unit in Mansoura University Hospital. The follicular fluid [FF] levels of sex steroids were assayed via high performance liquid chromatography. RNA was extracted from GC of all cases and controls. cDNA and amplification of a gene region from 1648 to 2055 bp of human androgen receptor was carried out by one step RT-PCR. The PCR product size of 400 bp was electrophoresed on 1% agarose gel. The results revealed that the F F levels of testosterone, androstenedione, 3alpha-androstanediol, 17 OH progesterone and progesterone were significantly higher in patients with PCO than the control group. Furthermore, there was a significant increase in AR expression in patients with PCO than the control group. It could be concluded that androgen signaling through AR plays an important role in GC development and is required for the optimal performance of female reproduction, but that excessive androgen signaling might lead to abnormal follicular growth seen in polycystic ovary syndrome

Concentração dos hormônios esteroides no fluido folicular de folículos ovarianos maduros e imaturos de pacientes com síndrome dos ovários policísticos submetidas à fertilização in vitro / Concentration of steroid hormones in the follicular fluid of mature and immature ovarian follicles of patients with polycystic ovary syndrome submitted to in vitro fertilization

OBJETIVO: avaliar a concentração dos hormônios esteroides no fluido folicular (FF) de folículos pequenos (10-14 mm) e grandes (> 18 mm) de mulheres com síndrome dos ovários policísticos (SOP) submetidas à hiperestimulação ovariana controlada (HOC) e ciclos de fertilização in vitro (FIV). MÉTODOS: estudo caso-controle foi conduzido em 13 mulheres inférteis com SOP (17 ciclos) e 31 mulheres inférteis por fator masculino - Grupo Controle (31 ciclos). Os FF foram aspirados individualmente e dividos em 4 grupos: G1 (FF pequeno do Grupo Controle), G2 (FF pequeno do grupo SOP), G3 (FF grande do Grupo Controle) e G4 (FF grande do grupo SOP). A metodologia utilizada para as dosagens de estradiol, progesterona e β-hCG foi a quimioluminescência, e de testosterona e androstenediona o radioimunoensaio. Para a análise das dosagens hormonais no FF entre os grupos SOP e Controle utilizou-se o teste t não-pareado, e para a comparação entre os quatro grupos, o ANOVA. Para a taxa de gravidez, foi utilizado o teste exato de Fisher. RESULTADOS: os folículos pequenos dos dois grupos tiveram valores menores de progesterona (8.435±3.305 ng/mL) comparados aos grandes (10.280±3.475 ng/mL), com valor de p<0,01. Os níveis de progesterona de todos os folículos do grupo SOP (8.095±4.151 ng/mL) foram inferiores ao Controle (9.824±3.128 ng/mL), com valor de p=0,03. Os níveis de testosterona diferiram entre G1 (326,6±124,4 ng/dL) e G3 (205,8±98,91 ng/dL), com valor de p<0,001, e entre G3 (205,8±98,91 ng/dL) e G4 (351,10±122,1 ng/dL), com valor de p<0,001. Os folículos pequenos (508,9±266 ng/dL) apresentaram valores superiores de testosterona comparados aos grandes (245,10±123 ng/dL), com valor de p<0,0001. As taxas de gravidez não diferiram entre os grupos SOP (5/13, 38,5 por cento) e Controle (9/31, 40,9 por cento), com valor de p=072. CONCLUSÕES: mulheres com SOP apresentam altas concentrações de testosterona no FF, independentemente do estágio de desenvolvimento folicular, e níveis de progesterona diminuídos, sugerindo que fatores parácrinos podem inibir sua secreção pelas células foliculares. As taxas de gravidez mostraram que o tratamento de HOC e FIV é uma boa opção para mulheres com infertilidade secundária à SOP.

PURPOSE: to evaluate the concentration of steroid hormones in follicular fluid (FF) of small (10-14 mm) and large (> 18 mm) follicles of women with polycystic ovary syndrome (PCOS) submitted to controlled ovarian hyperstimulation (COH) and in vitro fertilization (IVF) cycles. METHODS: a case-control study was conducted on 13 infertile women with PCOS (17 cycles) and 31 infertile women due to male factor - Control Group (31 cycles). FF was aspirated individually and divided into four groups: G1 (FF of small follicles of the Control Group), G2 (FF of small follicles of the PCOS group), G3 (FF of large follicles of the Control Group) and G4 (FF of large follicles of the PCOS group). Estrogen, progesterone and β-hCG were determined by chemiluminescence, and testosterone and androstenedione by radioimmunoassay. The unpaired t-test was used to compare the hormone determinations in the FF of the PCOS and Control Groups, and the four groups were compared by ANOVA. Fisher's exact test was used to compare the pregnancy rates. RESULTS: the small follicles of the two groups had lower progesterone levels (8,435±3,305 ng/mL) than large follicles (10,280±3,475 ng/mL), p-value <0.01. The progesterone levels of all follicles of group PCOS (8,095±4,151 ng/mL) were lower than Control (9,824±3,128 ng/mL), p-value =0.03. Testosterone differed between G1 (326.6±124.4 ng/dL) and G3 (205.8±98.91 ng/dL), p-value <0.001, and between G3 (205.8±98.91 ng/dL) and G4 (351.10±122.1ng/dL), p-value <0.001. Small follicles had higher testosterone levels (508.9±266 ng/dL) than large follicles (245.10±123 ng/dL), p-value <0.0001. The pregnancy rates did not differ between the PCOS (5/13, 38.5 percent) and the Control groups (9/31, 40.9 percent), p-value =072. CONCLUSIONS: women with PCOS had high testosterone concentrations in the FF, regardless of the stage of follicle development, and reduced progesterone levels, suggesting that paracrine factors may inhibit the secretion of the latter by follicular cells. The pregnancy rates showed that treatment with COH and IVF is a good option for women with infertility secondary to PCOS.

Effect of ovarian morphology on in vitro oocyte recovery and quality, and certain follicular fluid steroids hormones concentrations in ewes

The present study was conducted to investigate the effects of ovarian morphology on oocyte quantity and quality, as well as on follicular fluid steroid hormones concentrations. Fifty pairs of ovaries were collected from Barbari ewes and grouped into right, left, CL bearing and non-CL bearing ovaries. The weight, length, width and thickness of the right, left, CL bearing and non-CL bearing ovaries were recorded. The follicles were classified according to their diameter into 3 groups; small [<2mm], medium [2-4mm] and large [>4mm] follicles. Oocytes were classified according to their morphology into 3 grades; COCS [Compact cumulus oocyte complexes], POCS [Partially invested with less than three layers of cumulus cells] and DO [denuded oocyte]. The concentrations of progesterone and estradiol 17 beta in the follicular fluid were estimated. Results indicated that, dimensions of both right and left ovaries were not significantly differed. However, the ovarian dimensions as well as their weights were significantly [P < 0.05] affected by the presence of CL, being higher in the CL bearing ovary. The average number of large follicles were significantly [P < 0.05] increased in the right ovary when compared to the left one. The recovered COCs number was found to be significantly higher [P < 0.05] in the right than left ovaries. A greater number of vesicular follicles and aspirated COCS were found in the non-CL bearing ovary than in the CL bearing ovary. The non CL bearing ovaries provide larger numbers as well as higher quality of COCs when compared to CL bearing ovaries and that the former can be used to collect good quality COCs for in vitro production of sheep embryos. The progesterone concentration of follicular fluid was significantly higher in CL- and non-CL bearing ovaries [27.75 and 12.33 ng/ml; P < 0.05, respectively]. Non-CL bearing ovaries had significantly [P < 0.05] higher concentration of estradiol 17beta than those found in CL bearing ovaries [22.10 vs.8.43 pg/ml, respectively]. It can be concluded that non-CL bearing ovaries provide a higher number as well as superior quality of COCs than those obtained from ovaries bearing CL suggesting that the ovaries without CL can be used to collect good quality of COCs in view of in vitro production of sheep embryos [IVP]

MMP-9 in follicular fluid in PCO and non PCO females undergoing ICSI

Matrix Metalloproteinase-9 [MMP-9] is a proteolytic enzyme, the significance of which in the follicle maturation has been widely studied in non-stimulated animal models and humans. To compare the level of this enzyme in follicular fluid in PCO [polycystic ovary] and non PCO females undergoing intracytoplasmic sperm injection [ICSI]. MMP-9 was measured in the follicular fluid in 20 PCO and 30 non PCO females undergoing ICSI. Total MMP-9 showed no statistically significant difference between the two groups [p=0.145].While, MMP-9 per oocyte showed a statistically significant difference between the two groups, [p=<0.001]. So MMP-9 per oocyte is greater in non PCO group. There is no difference between MMP-9 secretion in follicular fluid in PCO and non PCO females undergoing ICSI

Effect of ovine follicular fluid peptide on ovarian responses and other organ weights in rats, Rattus norvegicus Berkenhout 1769.

The present study was aimed to study the effect of an ovine follicular fluid peptide on ovarian follicle and good oocyte numbers and weights of ovary, uterus, liver, pancreas and kidney in rats, R. norvegicus. A 30.1 kDa peptide was isolated from ovine follicular fluid by ammonium sulphate precipitation and then gel filtration. The peptide was tested at various levels in normal (22 and 36 day-old), superovulated (29 day-old) immature and 121-day old mature rats on the ovarian responses and other organ weights. The isolated peptide inhibited the growth of antral follicles in normal and superovulated rats. Ovarian, uterine weight and recovery of good oocytes were reduced when the peptide was administered at 100 microg dose. The peptide had no effect on kidney, liver, pancreas weight and recovery of preantral follicles.

Prolactin and cortisol levels in women with endometriosis

Endometriosis is a progressive estrogen-dependent disease affecting women during their reproductive years. The objective of the present study was to investigate whether endometriosis is associated with stress parameters. We determined cortisol and prolactin levels in serum, peritoneal and follicular fluid from infertile women with endometriosis and fertile women without the disease. The extent of the disease was staged according to the revised American Fertility Society classification (1997). Serum and peritoneal fluid were collected from 49 women aged 19 to 39 years undergoing laparoscopy. Eighteen women had stage I-II endometriosis and 10 had stage III-IV. Controls were 21 women undergoing laparoscopy for tubal sterilization. Follicular fluid was obtained from 39 women aged 25-39 years undergoing in vitro fertilization (21 infertile women with endometriosis and 18 infertile women without endometriosis). Serum prolactin levels were significantly higher in infertile women with stage III-IV endometriosis (28.9 ± 2.1 ng/mL) than in healthy controls (13.2 ± 2.1 ng/mL). Serum cortisol levels were significantly higher in infertile women with stage III-IV endometriosis (20.1 ± 1.3 ng/mL) than in controls (10.5 ± 1.4 ng/mL). Cortisol and prolactin levels in follicular fluid and peritoneal fluid did not differ significantly between groups. The high levels of cortisol and prolactin in the serum from women with endometriosis might contribute to the subfertility frequently associated with the disease. Moreover, since higher levels of cortisol and prolactin are often associated with stress, it is probable that stress might contribute to the development of endometriosis and its progression to advanced stages of the disease.

Identification of New Proteins in Follicular Fluid from Mature Human Follicles by Direct Sample Rehydration Method of Two-Dimensional Polyacrylamide Gel Electrophoresis

Human follicular fluid (HFF) includes various biologically active proteins which can affect follicle growth and oocyte fertilization. Thus far, these proteins from mature follicles in human follicular fluid have been poorly characterized. Here, two-dimensional polyacrylamide gel electrophoresis (2-DE) with matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to identify new proteins in HFF. Mature follicular fluids were obtained from five females after oocyte collection during in vitro fertilization (IVF). We directly rehydrated HFF samples, obtained high-resolution 2-DE maps, and processed them for 2-DE and MALDI-MS. One hundred eighty spots were detected and 10 of these spots were identified. By the 2-DE database, six of them had been reported, as proteins already existing in HFF. Hormone sensitive lipase (HSL), Unnamed protein product 1 (UPP1), Unnamed protein product 2 (UPP2), and apolipoprotein A-IV precursor were newly detected. HSL and apolipoprotein A-IV participate in lipid metabolism. UPP1 has a homology with selenocysteine lyase. We found by RT-PCR that these genes are expressed from human primary granulosa cells. The proteins identified here may emerge as potential candidates for specific functions during folliculogenesis, hormone secretion regulation, or oocyte maturation. Further functional analysis of these proteins is necessitated to determine their biological implications.

Estradiol and testosterone concentrations in follicular fluid as criteria to discriminate between mature and immature oocytes

The objective of the present study was to examine the association between follicular fluid (FF) steroid concentration and oocyte maturity and fertilization rates. Seventeen infertile patients were submitted to ovulation induction with urinary human follicle-stimulating hormone, human menopausal gonadotropin and human chorionic gonadotropin (hCG). A total of 107 follicles were aspirated after hCG administration, the oocytes were analyzed for maturity and 81 of them were incubated and inseminated in vitro. Progesterone, estradiol (E2), estrone, androstenedione, and testosterone were measured in the FF. E2 and testosterone levels were significantly higher in FF containing immature oocytes (median = 618.2 and 16 ng/ml, respectively) than in FF containing mature oocytes (median = 368 and 5.7 ng/ml, respectively; P < 0.05). Progesterone, androstenedione and estrone levels were not significantly different between mature and immature oocytes. The application of the receiver-operating characteristic curve statistical approach to determine the best cut-off point for the discrimination between mature and immature oocytes indicated levels of 505.8 ng/ml for E2 (81.0 percent sensitivity and 81.8 percent specificity) and of 10.4 ng/ml for testosterone (90.9 percent sensitivity and 82.4 percent specificity). Follicular diameter was associated negatively with E2 and testosterone levels in FF. There was a significant increase in progesterone/testosterone, progesterone/E2 and E2/testosterone ratios in FF containing mature oocytes, suggesting a reduction in conversion of C21 to C19, but not in aromatase activity. The overall fertility rate was 61 percent but there was no correlation between the steroid levels or their ratios and the fertilization rates. E2 and testosterone levels in FF may be used as a predictive parameter of oocyte maturity, but not for the in vitro fertilization rate.

The acrosomic reaction in stallion spermatozoa: inductive effect of the mare preovulatory follicular fluid

In the female genital tract, spermatozoa must undergo capacitation and acrosome reaction prior to fertilization. A number of factors may induce physiological acrosome reaction assayed in vitro. The aims of this study are to determine the inductive effect of the preovulatory follicular fluid on the sperm acrosomal status in the equine, once some characteristics of the follicular fluid during folliculogenesis had been evaluated. The spermatozoa were obtained from cauda epididymes of adult stallion. Follicular fluid was taken from mare ovarian follicles classified according to their diameter. In these fluids, total protein, progesterone, estradiol and osmolarity were determined. Afterwards, the effect of preovulatory follicular fluid (50) upon induction of the acrosomic reaction in stallion capacitated spermatozoa was assayed. Results show that during folliculogenesis the ratio progesterone/estrogen is below 1. In large preovulatory follicles, there is a sharp increase of progesterone, reaching a ratio progesterone/estrogen close to 4. Protein concentration and osmolarity increase together with follicular development, being osmolarity very high at the preovulatory stage. Follicular fluid--in vitro--increases the percentage of spermatozoa with acrosome reaction, maintaining high rates of vitality and motility. The characteristics of follicular fluid undergo dynamic changes during the folliculogenesis, such as steroid level, protein concentration and osmolarity. These events may play a role in the reproductive process in vivo, considering that in vitro the follicular fluid is a very effective inductor of the acrosome reaction, with optimum levels of vitality and motility.

[Serum and follicular fluid IL-6 and sex steroid hormone levels and their correlation of undergoing IVF-ET with endometriosis and pregnancy rate in women.]

Cytokines Not only do rcgulate physiological processes, but also play important roles in immunopathological reactions. The aim of this study was to evaluate the correlation between IL-6 and sex hormone levels with endometriosis and pregnancy rate at the time of oocyte retrieval. In infertile women undergoing IVF-ET Eighty patients received ovulation induction drugs and underwent IVF-ET. IL-6 levels in serum and follicular fluid [FF] were measured by ELISA and FSH. estradiol and progesterone by RIA. In 36 out of 80 patients embryos were transferred and pregnancy rate were evaluated after two weeks. The result showed no significant correlation between serum and FF levels of estradiol. progesterone and FSH with pregnancy rate in IVF patients. Similar levels of lL-6 in serum and FE of pregnant and non pregnant patients after embryo transfer were observed [P> 0.05]. However there was a significant correlation between FF levels of IL-6 in pregnant and non pregnant endometriosis patients [P<0.05]. These results indicate that increase of FE levels of IL-6 may be accompanied by pregnancy rate reduction in endometriosis patients

Concentraçöes plasmáticas e foliculares de IGF-I em pacientes submetidas a FIV / Folicular and plasm IGF-I levels in IVF patients

RETROSPECTIVA: diversos estudos têm atribuído ao IGF-I a atividade gonadotrófica sobre as células foliculares, exercendo um efeito parácrino na regulaçäo da esteroidogênese. Entretanto, a verdadeira origem do IGF-I encontrado no fluido folicular ovariano permanece obscura. OBJETIVOS: determinar a relaçäo entre as concentraçöes de IGF-I folicular e sérico em 22 pacientes submetidas a fertilizaçäo in vitro. METODOLOGIA: foram avaliadas as concentraçöes de IGF-I no plasma e no fluido folicular de 22 mulheres submetidas a fertilizaçäo in vitro. As amostras foram obtidas na ocasiäo da captaçäo de oócitos e dois dias antes, na ocasiäo da administraçäo de HCG. O IGF-I foi dosado em cada uma das amostras pelo método imunorradiométrico. RESULTADOS: houve correlaçäo dos níveis de IGF-I entre o plasma e o fluido folicular, sugerindo um provável equilíbrio dessa proteína entre os dois compartimentos. CONCLUSöES: esses achados säo condizentes com relatos prévios da literatura e sugerem que a maior parte do IGF-I presente no fluido folicular seja derivado de difusäo a partir do plasma e que a produçäo local desse fator de crescimento no interior do ovário seja pouco expressiva.

Funcion de distintas formas de fibronectina en el desarrollo de foliculos bovinos in vitro / Role of different forms of fibronectin in in vitro bovine follicular development

En el presente estudio, se analizó la posibilidad de que las distintas formas de fibronectina (FN), producidas como resultado de la maduración alternativa (alternative splicing) del mensajero, ejerzan funciones diferenciales en el desarrollo folicular. En particular se determinó la presencia de la región ED-I, ausente en la FN plasmática, tanto a nivel de mensajero como de proteína, durante este proceso. El análisis de los niveles de FN en fluidos foliculares correspondientes a distintas etapas de desarrollo mostró marcadas variaciones en la concentración de FN ED-I+ y los de permanecieron relativamente constantes. En folículos corespondientes a la fase de selección se observó una correlación inversa entre los niveles de FN ED-I+ y los de estradio (p<0.001). El tratamiento con estradiol no tuvo efecto sobre el splicing alternativo de FN en cultivos de células de la granulosa bovinas, mientras que el AMPc tuvo un efecto inhibitorio sobre la incorporación de ED-I. Por otra parte, el factor de crecimiento transformante tipo beta (TGF-beta) estimuló tanto la produción de FN total como la inclusión de la región ED-I. Este efecto fue verificado tanto a nivel de la proteína (Western blots) como del ARN mensajero (Northern blots). Un péptido correspondiente a la región ED-I tuvo un efecto estimulatorio sobre el crecimento de una línea de células de la granulosa bovinas (BGC-1) mientras que el péptido correspondiente e las regiones flanqueantes no tuvo efecto. Los datos presentados en este estudio plantean una nueva forma de regulación mediante la cual cambios cualitativos en la estructura primaria de la FN podrían mediar algunas de las acciones de gonadotrofinas y factores intraováricos durante el desarrollo folicular.

Implicancias del sistema GH (Growth Hormone) IGF-I (Insuline like growth factor 1) E IGFBP-3 (Insuline like growth factor binding protein-3) sobre la competencia ovocitaria en fertilización in vitro / Implicating of GH system, IGF-1 and IGFBP-3 on oocyte competence at in vitro fertilization

Objetivos: Investigar la asociación entre los niveles de GH - IGF-I -IGFBP-3 y progesterona (Pg) en líquido folicular humano y el nº de ovocitos reclutados, su madurez, fertilización y clivaje in vitro. Material y Métodos: Diseño: Observacional, analítico transversal. Se estudió el fluido folicular de 74 folículos, cada uno conteniendo un ovocito, en 17 pacientes, luego de su aspiración para FIV-ET, registrándose para cada ovocito su clasificación y evolución en cultivo. Se midieron los niveles de GH, IGF-I, IGFBP-3 y Pg por RIA. Los valores se expresan en Md (rango intercuartílico). Resultados: El grupo de pacientes de baja respuesta folicular (ovocitos < ó=3) presentó menores niveles intrafoliculares de GH y Pg que las normorespondedoras. [0,7 (0,34) vs 1,25 (0,87) ng/ml, p < 0,01; 6.775,1 (4.225,5) vs 10.180,0 (6.700,5) p < 0,01, respectivamente]. Las concentraciones intrafoliculares de GH e IGF-I fueron significativamente mayores en el grupo que recibió análogos GnRH [1,26 (1,12) vs 0,56 (0,74) ng/ml, p < 0,002 y 0,62 (0,3) vs 0,42 (0,34) U/ml, p < 0,03]. La fertilización ovocitaria y clivaje positivos se asociaron a mayores concentraciones de GH [1,26-(1,1) vs 0,56 (0,98) ng/ml, p < 0,02], IGF-1 [0,63 (0,28) vs 0,51 (0,3), p < 0,05] e IGFBP-3 [2,8 (1,4) vs 1,89 (1:91) p < 0,01]. Conclusiones: Los resultados observados sugieren: 1) Existiría una asociación entre la baja respuesta ovocitaria y menores concentraciones intrafoliculares de GH y Pg. 2) La administración de análogos modularía positivamente el sistema GH - IGF-I durante la foliculogenesis. 3) A su vez la exposición a mayores niveles de GH e IGF-I se vincularía a una myor habilidad del ovocito para fertilizar y clivar in vitro

Characteristics of proteoglycans of buffalo ovarian follicular fluid during maturation of follicles.

The proteoglycans (PGs) and glycosaminoglycans (GAGs) of buffalo ovarian follicular fluid (FF) have been studied in small (2-4.9 mm), medium (5-9.9 mm) and large (> or = 10 mm) follicles. GAGs in different categories of follicles were isolated, assayed and analysed. On the basis of hexosamine analysis, glucosamine accounted for all the free GAGs in FF of small and medium follicles. No free GAG was found in large follicles. The concentration of GAGs in the form of PGs decreased significantly with follicular maturation. Qualitative analysis of GAGs from PGs showed higher galactosamine than glucosamine. The ratios of GalNH2:GluNH2 and neutral sugars were highest in small follicles followed by medium and large follicles. On the other hand, the percentage of sialic acid in GAGs was highest in large follicles followed by medium and small follicles. The fractionation of PGs by gel filtration indicated the presence of two types of PGs in buffalo ovarian FF. Difference in distribution of two types of PGs in small and large follicles was also noted.

Some biochemical components of follicular fluid and its relation to ovum fertilization in IVF program

The objective of this study was to determine the importance of follicular fluid micro-environment for growth and maturation of oocytes and their relation to ovum fertilization in IVF program. The study was done in the Egyptian IVF and ET Center in Maadi. Twenty seven infertile females underwent an IVF program from the period of November 1992 to April 1993, patients were stimulated by [GnRh-HMG- HCG]. The follicular fluids for all patients were biochemically assayed for inorganic pyrophosphatase, alkaline phosphatase activity, cholesterol, high and low density lipoproteins [HDL and LDL], total and ionized calcium in ions, sodium, potassium and chlorides. The mean F.F. concentration of total calcium was 69.5 +/- 19.9 mg/ml, ionized calcium was 1.03 +/- 0.17 mmol/L, while alkaline phosphatase was 0.074 +/- 0.036 U/L and phosphatase was 0.347 +/- 0.393 U/ml. The correlation between the number of fertilized ova and the contents of all aspirated follicles studied, was highly significant with cholesterol [p < 0.005], HDL [p < 0.025], LDL [p < 0.05], ionized calcium [p < 0.025] and total calcium [p < 0.05] while the correlation was poor with alkaline phosphatase, pyrophosphatase, sodium, potassium and chloride. It was concluded that, F.F. Lipids have a direct correlation, although it is variable to fertilization rate in humans. Alkaline phosphatase, pyrophosphatase, sodium, potassium and chloride have no correlation to fertilization rate Document 11

Maduración del ovocito y su relación con algunos componentes del líquido folicular / Oocyte maturation and its relation with some follicular fluid components

Se analizaron 35 muestras de líquido folicular (LF) no hemático, obtenido de pacientes que recibieron inductores de ovulación múltiple para los protocolos de fertilización in vitro (FIV) y transferencia intratubaria de gametos (GIFT). El objetivo de este estudio retrospectivo, fue el de localizar uno o varios marcadores bioquímicos del estado de madurez del ovocito presentes en el LF, y correlacionalos con el aspecto morfológico del mismo. El grado de expanción del complejo ovocitocorona radiata-cumulus oofurus (COCC), sirvió para la primera clasificación de este. El líquido folicular fue sometido a barrido espectrofotométrico en el área de luz visible entre 350 a 600 nm para medir la absorbancia de su pigmento amarillo. Los niveles foliculares de proteínas estradiol y progesterona se midieron y correlacionaron con la morfología del COCC. La máxima absorbancia obtenida con el análisis espectrofotométrico, mostró una gama de picos que al ser agrupados y promediados formaron cuatro grupos principales: El primero formado por 20 muestras, con un pico promedio de 413.8 nm, el 45 por ciento de los ovocitos que formaron este grupo fueron clasificados como inmaduros de acuerdo al aspecto del COCC. El grupo dos con 7 muestras, presentó un pico a 457.2 nm, todos estos ovocitos fueron catalogados como maduros o preovulatorios. El grupo tres presentó una combinación de dos picos del primero a 416.4 nm y el segundo a 457.9 nm, con solo tres muestras de LF, con dos ovocitos clasificados como inmaduros y uno atrésico. El cuarto grupo con 5 LF, también presentó dos picos: el primero a 413.6 nm y otro a 562 nm, dos de estos ovocitos fueron clasificados como atrésicos y los restantes como preovulatorio. En este estudio no se encontró correlación entre los niveles de proteínas, estradiol y progesterona con la morfología del COCC. Se concluye que la presencia de estos picos de máxima absorbancia pueden ser de gran utilidad clínica para corroborar el estado de madurez del ovocito, sobre todo en los casos de GIFT en donde la transferencia de los mismos es inmediata, y se puede elegir con mayor seguridad los ovocitos maduros

Localization and detection of ovarian follicular fluid protein in follicles of human ovaries.

Human ovarian follicular fluid protein has been partially purified and the active fraction designated as hGF2. Using specific polyclonal antiserum to hGF2, it was observed to be localized immunohistochemically in the granulosa cells of medium but not large follicles of human ovary. The hGF2 levels were estimated by ELISA in serum and follicular fluid of 10 gonadotropin-stimulated women recruited for IVF-ET programme. The results revealed a 3-fold increase in the concentration of hGF2 in follicular fluid compared to that in serum of these patients. These data indicate that the protein is secreted by granulosa cells and plays an important role in the regulation of follicular maturation and ovulation.