An in silico model using prognostic genetic factors for ovarian response in controlled ovarian stimulation: A systematic review.

PURPOSE: To study the use of in silica model to better understand and propose new markers of ovarian response to controlled ovarian stimulation before IVF. METHODS: A systematic review and in silica model using bioinformatics. After the selection of 103 papers from a systematic review process, we performed a GRADE qualification of all included papers for evidence-based quality evaluation. We included 57 genes in the silica model using a functional protein network interaction. Moreover, the construction of protein-protein interaction network was done importing these results to Cytoscape. Therefore, a cluster analysis using MCODE was done, which was exported to a plugin BINGO to determine Gene Ontology. A p value of < 0.05 was considered significant, using a Bonferroni correction test. RESULTS: In silica model was robust, presenting an ovulation-related gene network with 87 nodes (genes) and 348 edges (interactions between the genes). Related to the network centralities, the network has a betweenness mean value = 102.54; closeness mean = 0.007; and degree mean = 8.0. Moreover, the gene with a higher betweenness was PTPN1. Genes with the higher closeness were SRD5A1 and HSD17B3, and the gene with the lowest closeness was GDF9. Finally, the gene with a higher degree value was UBB; this gene participates in the regulation of TP53 activity pathway. CONCLUSIONS: This systematic review demonstrated that we cannot use any genetic marker before controlled ovarian stimulation for IVF. Moreover, in silica model is a useful tool for understanding and finding new markers for an IVF individualization. PROSPERO: CRD42020197185.

Evaluation of three-dimensional SonoAVC ultrasound for antral follicle count in infertile women: its agreement with conventional two-dimensional ultrasound and serum levels of anti-Müllerian hormone.

BACKGROUND: Several studies have reported a correlation between antral follicle count by conventional 2D transvaginal sonography and serum anti-Müllerian hormone levels. However, few studies have investigated the effectiveness of 3D SonoAVC transvaginal ultrasound technology, particularly in infertile women. Therefore, this study aims to evaluate the usefulness of three-dimensional (3D) SonoAVC transvaginal ultrasound technology for antral follicle count and its correlation to conventional two-dimensional (2D) transvaginal ultrasound and serum levels of anti-Müllerian hormone in infertile women. METHODS: This cross-sectional study included 42 infertile women with age lower than 40 years that underwent treatment at a private fertility clinic between June and December 2015. Patient data included age, body mass index and cause of infertility. On cycle day 3 the following hormone levels were measured: serum levels of anti-Müllerian hormone, follicle-stimulating hormone, cancer antigen 125, prolactin, thyroid-stimulating hormone and oestradiol; the number of antral follicles was counted as well. The scanning were performed through 2D and 3D technology transvaginal ultrasound. RESULTS: Using a Bland-Altman test we demonstrated that both technologies are quite equivalent. However, antral follicle count is higher using 3D ultrasound technology compared to 2D technology (p < 0.001; Wilcoxon test), this finding is mainly remarkable in ovaries with more than 20 antral follicles. Moreover, the mean time required for manual 2D ultrasound and 3D SonoAVC measurements were 275 ± 109 and 103 ± 57 s, respectively (p < 0.001). Serum AMH concentration correlated to the total number of early antral follicles (correlation coefficients = 0.678 and 0.612; p < 0.001 by 2D ultrasound and 3D SonoAVC, respectively; Spearman's correlation test). CONCLUSIONS: Antral follicle count is better estimated using 3D ultrasound compared to 2D technology. A great advantage of 3D SonoAVC was less time required for an examination and the visual advantage when it need to count more than 20 follicles. TRIAL REGISTRATION: CAAE: 35141114.4.0000.5327 . Registered 10 June 2015.

Follicular-fluid anti-Müllerian hormone concentration is similar in patients with endometriosis compared with non-endometriotic patients.

This cross-sectional prospective study assessed follicular-fluid anti-Müllerian hormone (AMH) concentrations in infertile patients with mild/minimal endometriosis during natural IVF. Thirty-two women participated in the study. Patients were divided into two groups: tubal obstruction without endometriosis (control group) and minimal/mild endometriosis (study group). All patients underwent laparoscopy for assessment of infertility; at the same time, any foci of endometriosis found were cauterized. AMH concentration was measured in the follicular fluid of a single follicle when it achieved pre-ovulatory maturation by ultrasensitive enzyme-linked immunosorbent assay. Likewise, AMH, FSH and inhibin B content in serum was also measured. Age (30 ± 1.3 and 32 ± 0.8 years) and body mass index (22 ± 0.6 and 22 ± 0.5 kg/m(2)), day-3 antral follicle count (11.3 ± 1.7 and 10.7 ± 1.5), serum FSH concentrations (5.4 ± 0.6 and 5.0 ± 0.3 IU/ml) and follicular-fluid AMH concentrations (1.8 ± 0.3 and 1.5 ± 0.1 ng/ml, study and control group, respectively; mean difference 0.33, 95% CI -0.21 to 0.88) were similar in both groups. This study shows that infertile patients with minimal/mild endometriosis have a similar concentration of AMH in the follicular fluid after natural IVF as compared with control subjects.

Variability of endometrial glandular opening count in infertile patients prior to first IVF treatment.

The aim of the present study was to evaluate the number of endometrial glandular openings, using previously reported software that provides an objective count, and to assess the variability of this parameter during the luteal phase in a population of women who had no hormonal abnormalities presenting with tubal infertility or male factor infertility. A cross-sectional study was performed comprising 561 patients selected for a diagnostic hysteroscopy for the investigation of infertility. Hysteroscopy was performed during the mid-secretory phase prior to the first IVF treatment cycle. A total of 561 image frames from all patients were analysed. All images were automatically selected by the software, which also evaluated the number of endometrial glandular openings. The mean +/- SD glandular opening count was 53.2 +/- 30 (range 4-158). The analysis of variation showed a significant difference (P = 0.001) among all video frames. In conclusion, endometrial glandular opening count, as measured by the method described, can be used in investigations during the luteal phase. Although a lack of pattern was observed in endometrial maturation, this feature should be explored further in this subgroup of patients.

Sperm recovery prediction in azoospermic patients using Doppler ultrasonography.

PURPOSE: To evaluate power Doppler ultrasonography to predict sperm recovery in azoospermic patients. METHODS: Color Doppler and power Doppler ultrasonography of testis were performed in 38 patients before testicular sperm extraction. Analysis of blood flow included the pulsatility and resistance index of intratesticular vessels and testicular artery, and power Doppler of testis. The results of power Doppler of testis were classified into three categories: 0, no vessels found; 1, one to three vessels; 2, more than three vessels found. RESULTS: Power Doppler of both testis showed a significant difference between obstructive azoospermia and non-obstructive azoospermia (Fisher's exact test - P = 0.02), and between the groups with and without sperm recovery (Fisher's exact test - P = 0.001). Doppler indices of intratesticular vessels and testicular artery were similar between the groups. CONCLUSIONS: Testicular Power Doppler assessment showed that patients with obstructive azoospermia have better blood flow than patients with non-obstructive azoospermia, and power Doppler is able to predict sperm recovery in azoospermic patients.

Vaginal versus oral E(2) administration: effects on endometrial thickness, uterine perfusion, and contractility.

OBJECTIVE: To compare the effects of vaginal or oral E(2) administration on endometrial thickness, uterine perfusion, and contractility. DESIGN: Prospective, randomized, crossover study. SETTING: Assisted Reproduction Unit, Clamart, France. PATIENT(S): Thirty-nine infertile women undergoing 78 E(2)/P cycles. INTERVENTION(S): Women received micronized 17beta-E(2), 2 mg/day orally (cycle days 1 to 28) and P, 300 mg/day vaginally (cycle days 15 to 28). After a menstrual cycle washout interval, women received a similar treatment except that 17beta-E(2) was administered vaginally. MAIN OUTCOME MEASURE(S): Endometrial thickness, mean uterine artery pulsatility index, endometrial blood flow, and uterine contraction frequency assessed in ultrasound scans on cycle days 14 and 18. RESULT(S): On day 14, the endometrium was thicker (8.7 +/- 0.6 vs. 7.1 +/- 0.3 mm, P< .0001), pulsatility index values were lower (2.4 +/- 0.1 vs. 3.0 +/- 0.2, P< .0002), and endometrial blood flow tended to be increased in the vaginal E(2) cycles as compared to the oral E(2) cycles. On day 18, similar differences remained. However, P-induced decrease in contraction frequency was slighter in vaginal E(2) cycles (33% vs. 18%, P< .0003). CONCLUSION(S): Vaginal E(2) administration improves endometrial proliferation and uterine perfusion, presumably because of combined local and systemic effects, but may interfere with P-induced uterine relaxation.

ATP diphosphohydrolase activity in synaptosomes from cerebral cortex of rats subjected to chemically induced phenylketonuria.

ATP diphosphohydrolase (apyrase) (EC 3.6.1.5) activity was measured in synaptosomes from cerebral cortex of Wistar rats of both sexes subjected to experimental phenylketonuria, i.e., chemical hyperphenylalaninemia induced by subcutaneous administration of 5.2 mumol phenylalanine/g body weight (twice a day) plus 0.9 mumol p-chlorophenylalanine/g body weight (once a day). ATP diphosphohydrolase specific activity (nmol Pi min-1 mg protein-1) of synaptosomes was significantly decreased compared to controls for both ATP (from 147.6 to 129.9) and ADP (from 70.2 to 63.1) hydrolysis one hour after single administration of the drugs to 35-day old rats. Chronic treatment was performed from the 6th to the 28th postpartum day. The enzyme specific activity of synaptosomes was measured one week after the last administration of the drugs and was significantly reduced compared to controls for both ATP (from 164.1 to 150.2) and ADP (from 76.3 to 62.1) hydrolysis. The in vitro effects of the drugs on the synaptosome enzyme specific activity were also investigated. Phenylalanine alone or associated with p-chlorophenylalanine significantly reduced enzyme specific activity for both ATP (from 150.2 to 136.0) and ADP (from 70.5 to 59.3) nucleotides as substrates. Since ADP diphosphohydrolase seems to play an important role in neurotransmission, these findings may be related to the neurological dysfunction characteristic of human phenylketonuria.

Predictive factors for motile sperm recovery using testicular biopsy in nonobstructive azoospermic patients.

OBJECTIVE: The objective of the present study was to assess the predictive value of hormone values, histological analysis for the finding of motile spermatozoa on testicular biopsy in nonobstructive azoospermia. METHODS: This cross-sectional study assessed serum follicle-stimulating hormone (FSH), luteinizing hormone, prolactin and total testosterone values in 50 patients undergoing testicular biopsy prior to ICSI. P < 0.05 was considered significant. RESULTS: Recovery was successful in 28 cases, and motile spermatozoa were retrieved in 7. Significant differences were detected between presence of motile spermatozoa and absence of spermatozoa in terms of FSH values (P = 0.003, one-way ANOVA). The other variables did not present statistical differences. A receiver operating characteristic curve showed that FSH levels below 17.00 IU/L were predictive of motile spermatozoa recovery. CONCLUSIONS: FSH values can serve as a predictive factor for the recovery of motile spermatozoa using biopsy in azoospermic patients. On the other hand, histological analysis and other hormone values were not helpful in the prediction of motile sperm.

[Hysterosonosalpingography and hysterosalpingography in the diagnosis of tubal patency in infertility patients]. / Histerossonossalpingografia e histerossalpingografia no diagnóstico de permeabilidade tubária em pacientes inférteis.

OBJECTIVES: To compare the hysterosonosalpingography(HSS) to the hysterosalpingography(HSG) in the evaluation of the tubal factor in infertility patients. MATERIAL AND METHODS: A transversal study with 30 patients in infertility investigation, with age minor than 38 years was performed. The patients were submitted to HSS, HSG and laparoscopy (LPC) in the first cycle menstrual phase. The evaluation of tubal patency of HSS and HSG, with LPC like gold-standard was compared. It was calculated the negative predictive value (PV-) of the exams. The HSS used Ecovist like contrast agent, the HSG used water soluble contrast media and the LPC used tubal insufflation with methylene blue. It was considered significative a p< 0.05 RESULTS: The final sample was of 26 patients (4 leave the investigation). The mean age was of 30,6 years. The HSS showed tubal patency in at least one of the tubes in 24 patients (92,3%). The HSG showed tubal patency in one of the tubes in 25 patients (96,2%), and the LPC in 25 patients (96,2%). The PV- of the HSS was of 92% and the PV- of HSG was of 100%. The differences were not statistically significant (p=0.996, Yates test). CONCLUSIONS: The HSS is an alternative method in the evaluation of tubal patency in infertility patients.

TP53 PIN3 and PEX4 polymorphisms and infertility associated with endometriosis or with post-in vitro fertilization implantation failure.

p53 has a crucial role in human fertility by regulating the expression of leukemia inhibitory factor (LIF), a secreted cytokine critical for blastocyst implantation. To examine whether TP53 polymorphisms may be involved with in vitro fertilization (IVF) failure and endometriosis (END), we have assessed the associations between TP53 polymorphism in intron 2 (PIN2; G/C, intron 2), PIN3 (one (N, non-duplicated) or two (D, duplicated) repeats of a 16-bp motif, intron 3) and polymorphism in exon 4 (PEX4; C/G, p.P72R, exon 4) in 98 women with END and 115 women with post-IVF failure. In addition, 134 fertile women and 300 women unselected with respect to fertility-related features were assessed. TP53 polymorphisms and haplotypes were identified by amplification refractory mutation system polymerase chain reaction. TP53 PIN3 and PEX4 were associated with both END (P=0.042 and P=0.007, respectively) and IVF (P=0.004 and P=0.009, respectively) when compared with women both selected and unselected for fertility-related features. Haplotypes D-C and N-C were related to higher risk for END (P=0.002, P=0.001, respectively) and failure of IVF (P=0.018 and P=0.002, respectively) when compared with the Fertile group. These results support that specific TP53 haplotypes are associated with an increased risk of END-associated infertility and with post-IVF failure.

Peritoneal and serum interleukin-18 levels are not increased in women with minimum or mild endometriosis.

Interleukin-18 (IL-18) is a cytokine that belongs to the IL-1 family. Endometriosis is strongly associated with sub-fertility, and affects about 15% of women of reproductive age. IL-18 may favor the progression of endometriosis. The objective of the present study was to determine the concentration of IL-18 in the serum and peritoneal fluid of infertile women with endometriosis. Forty infertile and 25 fertile women were screened in a teaching hospital. Thirty-four infertile patients with minimal or mild endometriosis and 22 fertile controls were enrolled in the study. The primary outcome was the estimate of IL-18 levels and the secondary outcome was the correlation between serum and peritoneal levels of IL-18. There were no differences between the two groups regarding age, body mass index and levels of peritoneal fluid IL-18 (mean +/- SD): 290.85 +/- 173.02 pg/mL for infertile women vs 374.21 +/- 330.15 pg/mL for controls; or serum IL-18: 391.07 +/- 119.71 pg/mL for infertile women vs 373.42 +/- 129.11 pg/mL for controls. However, a positive association was found between serum and peritoneal IL-18 levels in patients with endometriosis: r = 0.794, P = 0.0001. All measurements were carried out at the same time by the Human IL-18 Immuno Assay ELISA kit (MBL Co. Ltd., Japan). The present study did not find evidence supporting the hypothesis that IL-18 levels are associated with infertility in women with minimal and mild endometriosis, although a positive correlation was detected in these women between peritoneal and serum levels of IL-18.

Serum anti-mullerian hormone levels and follicular cohort characteristics after pituitary suppression in the late luteal phase with oral contraceptive pills.

BACKGROUND: Anti-mullerian hormone (AMH) is a marker of ovarian function and reserve and reflects the number and size of antral follicles. The objective of this study was to evaluate the effect of FSH suppression on AMH levels, during the late luteal phase of human menstrual cycle, with the use of oral contraceptives pills (OCP). METHODS: Twenty normovulatory infertile women were included in the study. On the third day of a spontaneous menstrual cycle, the patients were submitted to a transvaginal ultrasound examination and blood sample collection. From the 20th day of this menstrual cycle, the patients took daily OCP, containing 0.030 mg of ethinyl-estradiol plus 0.15 mg of desogestrel. On the third day of the following cycle, the measurements were repeated. RESULTS: After OCP use, the levels of FSH and estradiol were significantly reduced (P < 0.001). The number of antral follicles measured on both occasions did not differ, although after OCP use, the follicles presented significantly lower diameters (mean 4.4 + 1.7 mm before OCP versus 3.5 + 1.2 mm after OCP P < 0.001). The levels of AMH were significantly reduced after pituitary suppression, with a median (inter-quartile range) of 3.02 ng/mL (1.21-6.39) before OCP and 2.22 ng/mL (0.9-3.11) after OCP, P = 0.04. CONCLUSIONS: The short administration of OCP in late luteal phase caused suppression of FSH secretion during the cycle transition, leading to a more homogeneous follicular cohort. The lower AMH levels observed, although simultaneous with FSH suppression, were probably not a direct effect of the reduced FSH levels, but were more likely a consequence of the lower production by the arrested follicular cohort.

Luteal phase oestradiol administration in ovarian stimulation cycles with GnRH antagonist is comparable to the GnRH agonist (long) protocol.

PURPOSE: The purpose of the study was to compare the effectiveness of GnRH antagonist with luteal phase estradiol administration to GnRH agonist cycles, long protocol. METHODS: 55 IVF-ICSI patients received oestradiol in the luteal phase of the cycle, before a cycle with GnRH antagonist. Fifty-five patients submitted to IVF-ICSI with the use of agonist were allocated, age matched, as a control group (historical control). The primary outcome was the number of retrieved oocytes. RESULTS: Patients were similar in terms of clinical characteristics. No differences were found in the number of oocytes retrieved (study group, 8.1 +/- 4.7; control group, 7.4 +/- 4.5) or in oocyte quality. CONCLUSIONS: We clearly demonstrated that the effectiveness of GnRH antagonist when combined with luteal phase estradiol is comparable to GnRH agonist cycles.

Subendometrial power Doppler quantification: a new classification proposal.

Fifty-two women with regular menses were enrolled in the study. The patients were not allowed to use non-steroidal anti-inflammatory drugs within 24 h of any examination. All patients were examined during the mid-luteal phase (6-9 days after ovulation, according to previous ultrasound record). Power Doppler energy levels were classified into five categories according to the per cent area of sub-endometrial signal: I (<10%), II (10-25%), III (25-50%), IV (50-75%) and V (>75%). The colour Doppler signal was considered positive when it reached at least the endometrial basal layer. The picture of the endometrium was analysed and the regions of interest were identified and marked for further analysis. Each recorded image was then independently evaluated and classified by three blinded observers. According to the power Doppler classification, age, body mass index (BMI) and endometrial thickness were analysed, and no significant differences were observed among them. The Kappa test (0.70) demonstrated an excellent agreement among examiners (P = 0.0001). This study has validated a very simple and cost-effective classification for sub-endometrial vascularization. This method of quantification may potentially be of use, and its relevance to clinical practice should be explored.

Insulin-like growth factor-1 and insulin-like growth factor binding protein-1 and 3 in the follicular fluid of infertile patients submitted to in vitro fertilization.

PURPOSE: In the present article we propose to evaluate IGF-1, IGFBP-1 and 3 in the follicular fluid of infertile patients submitted to in vitro fertilization. METHODS: We performed a case-control study with 53 infertile patients submitted to the first in vitro fertilization attempt. We compared their follicular fluid concentration of IGF-1, IGFBP-1 and IGFBP-3 between the patients who became pregnant (n = 11) versus those nonpregnant (n = 42). RESULTS: The clinical characteristics of patients from the two groups were similar in terms of age and body mass index. Data related to the analysis of ovulation induction was not different regarding length of induction in days, number of retrieved oocytes, fertilization rate, and number of transferred embryos. Furthermore, the number of FSH units required for ovarian induction was also similar between the studied groups. IGF-1 and IGFBP-1 were not significantly different between the groups (p > 0.05). However, those patients that became pregnant presented a lower follicular fluid concentration of IGFBP-3, 2237.10 +/- 582.73 pg/ml and 2657.64 +/- 584.15 ng/ml, respectively (p = 0.038). CONCLUSIONS: We demonstrated an association of a lower follicular fluid IGFBP-3 in individuals that became pregnant compared to subjects that did not after in vitro fertilization.

The use of GnRH antagonists in ovarian stimulation.

GnRH antagonists induce a rapid decrease in LH and FSH, preventing and interrupting LH surges. Their properties do not require a desensitization period, and this allows their use in the late follicular phase. GnRH antagonists could replace GnRH agonists in controlled ovarian stimulation without their side-effects and their long desensitization period. Two protocols for assisted reproduction technology (ART) cycles were designed: the single-dose protocol allies simplicity and efficacy, while the multiple-dose protocol is efficient and could reduce monitoring of the cycle, though compliance is mandatory. A review of the available literature on GnRH antagonists in ART cycles is presented, focusing on phase III controlled trials and ART results. Both protocols using GnRH antagonists were associated with the need for a smaller dose of gonadotrophin, a shorter stimulation period and a lower incidence of ovarian hyperstimulation syndrome (OHSS), albeit with statistically comparable pregnancy rates. A trend is observed in all studies showing a lower pregnancy rates in GnRH antagonist cycles as compared with GnRH agonist cycles. The role of the lower number of embryos, and the potential adverse effects of GnRH antagonists on endometrium or follicle must be studied. More cycles using GnRH antagonists are necessary to confirm their equivalent pregnancy rates. There is room for improvement in both protocols with regard to scheduling, antagonist dose level and the timing of its administration. Until further studies have been conducted, luteal support seems to remain mandatory. Perinatal outcome appears similar to that with other stimulation regimens. Triggering of ovulation can be obtained with GnRH agonist for patients at risk of OHSS. With regard to GnRH antagonists, questions remain regarding pregnancy rates, the indications of their use in patients with polycystic ovary syndrome or poor responders, and in ovarian stimulation outside IVF.

Insulin-like growth factor (IGF)-1 and IGF binding protein-1 and -3 in the follicular fluid of infertile patients with endometriosis.

BACKGROUND: Endometriosis is associated with pituitary-ovarian axis dysfunction. The study of the follicular fluid in patients with endometriosis is important to elucidate the pathophysiological mechanism of this disease. The objective of this present paper was to analyse the dosages of insulin-like growth factor-1 (IGF-1) and IGF binding protein-1 and 3 (IGFBP-1 and IGFBP-3) in the follicular fluid environment of infertile patients with endometriosis. METHODS: A total of 41 infertile patients undergoing IVF between January 1999 and January 2000 participated in the cross-sectional prospective study. Patients were divided into three groups: group I, minimal/mild endometriosis (n = 12); group II, moderate/severe endometriosis (n = 10); and group III, tubal obstruction (n = 19). The ultra-short protocol was used in association with recombinant FSH for ovulation induction. Follicular fluid analysis was performed using radioimmunoassay with specific kits. RESULTS: Follicular fluid IGF-1 and IGFBP-3 levels were not significantly different among the groups; however, follicular fluid IGFBP-1 levels were lower in those patients with moderate/severe endometriosis (P < 0.05). Comparison of ovulation induction time, number of recombinant FSH units, number of follicles, oocytes and embryos, and fertilization and gestation/cycle rates showed non-significant differences. CONCLUSION: Infertile patients with moderate/severe endometriosis, which is associated with ovulatory dysfunction, presented lower levels of IGFBP-1 in the follicular fluid when undergoing IVF.

Growth hormone and prolactin secretion after metoclopramide administration (DA2 receptor blockade) in fertile women.

In this report, we will describe the results of a cross-sectional study to assess PRL and GH secretion during the early follicular phase in 22 fertile patients after metoclopramide administration in order to achieve a dopaminergic DA2 receptor blockade. Blood samples were collected at - 15, 0, 15, 30, 45 and 60 minutes. PRL, GH, estradiol, IGF-I, TSH, glucose, and insulin were measured in the samples taken at - 15 and 0 minutes. The existence of a correlation between GH and PRL secretion was investigated. All patients presented normal serum levels of estradiol, prolactin, insulin, fasting glucose and IGF-I. Serum GH levels were not changed after metoclopramide infusion (p = 0.302), but there was a significant alteration in serum PRL (p = 0.0001) with the highest levels after 30 (mean: 237.20 ng/ml +/- 95.86) and 45 (mean: 211.80 ng/ml +/- 83.24) minutes. Serum GH levels did not correlate with serum PRL levels after the dopaminergic DA2 blockade. We conclude that GH secretion was not modulated by a direct effect of type 2 dopamine receptor.

Hyperprolactinemia and luteal insufficiency in infertile patients with mild and minimal endometriosis.

The objective of the present paper was to assess the presence of hormonal alterations in infertile women with stage I or II endometriosis (Group III, n = 20) compared to fertile women without endometriosis (Group I, n = 14) and to fertile women with endometriosis (Group II, n = 7). Serum levels of FSH, LH, estradiol, TSH, and PRL were measured between days 1 and 5 of the early follicular phase; in the luteal phase, three serum samples were collected for progesterone measurement, and endometrial biopsies were performed. Serum estradiol levels were lower (p = 0.035) in infertile patients with endometriosis than in fertile patients without endometriosis. Six infertile patients with endometriosis presented prolactin levels above 20 ng/ml. This was not observed in the other groups. Luteal insufficiency was more frequent in infertile patients with endometriosis (78.9%) than in fertile patients with (42.9%) or without endometriosis (0%). In a multiple logistic regression analysis, only the presence of endometriosis and infertility was significantly associated with luteal insufficiency. The serum levels of LH, FSH, and TSH were not significantly different among the groups. Luteal insufficiency and altered prolactin secretion were associated with endometriosis, and could be important mechanisms causing infertility in this group of patients.

Effect of phenylalanine and its metabolites on ATP diphosphohydrolase activity in synaptosomes from rat cerebral cortex.

The in vitro effects of phenylalanine and some of its metabolites on ATP diphosphohydrolase (apyrase, EC 3.6.1.5) activity in synaptosomes from rat cerebral cortex were investigated. The enzyme activity in synaptosomes from rats subjected to experimental hyperphenylalaninemia (alpha-methylphenylalanine plus phenylalanine) was also studied. In the in vitro studies, a biphasic effect of phenylalanine on both enzyme substrates (ATP and ADP) was observed, with maximal inhibition at 2.0 mM and maximal activation at 5.0 mM. Inhibition of the enzyme activity was not due to calcium chelation. Moreover, phenylpyruvate, when compared with phenylalanine showed opposite effects on the enzyme activity, suggesting that phenylalanine and phenylpyruvate bind to two different sites on the enzyme. The other tested phenylalanine metabolites phenyllactate, phenylacetate and phenylethylamine) had no effect on ATP diphosphohydrolase activity. In addition, we found that ATP diphosphohydrolase activity in synaptosomes from cerebral cortex of rats with chemically induced hyperphenylalaninemia was significantly enhanced by acute or chronic treatment. Since it is conceivable that ATPase-ADPase activities play an important role in neurotransmitter (ATP) metabolism, it is tempting to speculate that our results on the deleterious effects of phenylalanine and phenylpyruvate on ATP diphosphohydrolase activity may be related to the neurological dysfunction characteristics of naturally and chemically induced hyperphenylalaninemia.