Anti-mullerian hormone is the best predictor of poor response in ICSI cycles of patients with endometriosis

Purpose: To correlate ovarian reserve (OR) markers with response in assisted reproduction techniques (ART) and determine their ability to predict poor response among patients with endometriosis (EDT). Methods: We evaluated ART cycles of 27 women with EDT and 50 with exclusive male factor. Basal follicle stimulating hormone (FSH) and anti-mullerian hormone (AMH) levels were determined. Ovarian response to gonadotropin stimulation was assessed and correlation coefficients calculated between the variables and reserve markers. Areas under the curve (AUC) determined ability of tests to predict poor response. Results: AMH was significantly correlated with response in both groups and it was the only marker with significant discriminative capacity to predict poor response among EDT (AUC = 0.842; 95% CI: 0.651-0.952) and control group (AUC = 0.869; 95% CI: 0.743-0.947). Conclusion: Infertile patients with endometriosis can benefit from the pre-therapeutic assessment of OR markers. However, regardless of disease presence, only AMH predicts poor response to stimulus.

Increased basal FSH levels as predictors of low-quality follicles in infertile women with endometriosis

Objective: To determine whether basal levels of follicle-stimulating hormone (FSH) and anti-mullerian hormone (AMH), antral follicle count (AFC), and the numbers of dominant follicles, oocytes, and mature oocytes retrieved after ovarian stimulation differed between infertile women with endometriosis and healthy women undergoing assisted reproduction techniques (ART). Method: Of 77 consecutive ART candidates, 27 were infertile and had endometriosis. A male factor caused the infertility of the other 50, who acted as controls. Results: The AMH and AFC levels were similar in the 2 groups. The FSH levels were higher (8.28 mIU/ML [range, 5.25-24.1 mIU/ML] vs 5.91 mIU/mL [range, 2.47-18.7 mIU/ML]; P<0.01) in the study group. And the numbers of retrieved (n = 5 [range, 0-12] vs n = 9 [range, 0-27]; P<0.05) and mature oocytes (n = 4 [range, 0-11] vs n = 5 [range, 0-16]; P<0.05) were less in the study group. Conclusion: Because AMH levels were unchanged, endometriosis seems not to damage the primordial pool of follicles and oocytes, but to lessen the quality of the ovarian response to the hCG injection. Basal FSH levels may be of value in predicting ART success in women with the disease. (C) 2010 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

Bispectral monitoring in dogs subjected to ovariohysterectomy and anesthetized with halothane, isoflurane or sevoflurane

Objective To assess the effect of halothane (H), isoflurane (I) or sevoflurane (S) on the bispectral index (BIS), and the effect of the addition of meperidine in dogs subjected to ovariohysterectomy. Study design Prospective, randomized, blinded, clinical trial. Animals Forty-eight female mixed-breed dogs, with weights varying from 10 to 25 kg. Methods All dogs were premedicated with acepromazine (A) (0.1 mg kg(-1) IM) or A and meperidine (M) (3 mg kg(-1) IM) and they were divided into six groups of eight animals (AH, AMH, AI, AMI, AS, and AMS). Fifteen minutes after premedication they were anesthetized with propofol (5 mg kg(-1) IV) and then orotracheally intubated. Anesthesia was maintained with halothane, isoflurane or sevoflurane, respectively. The BIS, E`(anest) variables were recorded at 15 minutes after administering pre-anesthetic medication (T0); 10 minutes of anesthesia maintenance (T1); right ovarian pedicle ligation (T2); muscle suturing (T3); skin suture (T4) and 10 minutes after terminating the inhalant anesthetic (T5), respectively. Results BIS values were decreased at all times when compared to the baseline values in all groups (p < 0.05). In the comparative assessment between groups, the values obtained at T0 and T1 were similar for all groups. At T2, the values in AMH were lower than those obtained in AI, AMI and AS (p < 0.05). At the same time significantly higher values were found for AI when compared to AMS (p < 0.01). There was a correlation between the bispectral index and the expired anesthetic fraction in all groups. Conclusions and clinical relevance Within groups given the same inhalant anesthetic the bispectral index was a good indicator for the degree of hypnosis in dogs, indicating a good correlation with the amount of anesthetic and the nociceptive stimulation. BIS was a less reliable indicator of relative anesthetic depth when comparing equipotent end-tidal concentrations between the three inhalants.

Novel genes regulated by Sonic Hedgehog in pluripotent mesenchymal cells

Sonic Hedgehog is a secreted morphogen involved in patterning a wide range of structures in the developing embryo. Disruption of the Hedgehog signalling cascade leads to a number of developmental disorders and plays a key role in the formation of a range of human cancers. The identification of genes regulated by Hedgehog is crucial to understanding how disruption of this pathway leads to neoplastic transformation. We have used a Sonic Hedgehog (Shh) responsive mouse cell line, C3H/10T1/2, to provide a model system for hedgehog target gene discovery. Following activation of cell cultures with Shh, RNA was used to interrogate microarrays to investigate downstream transcriptional consequences of hedgehog stimulation. As a result 11 target genes have been identified, seven of which are induced (Thrombomodulin, GILZ, BF-2, Nr4a1, IGF2, PMP22, LASP1) and four of which are repressed (SFRP-1, SFRP-2, Mip1-gamma, Amh) by Shh. These targets have a diverse range of putative functions and include transcriptional regulators and molecules known to be involved in regulating cell growth or apoptosis. The corroboration of genes previously implicated in hedgehog signalling, along with the finding of novel targets, demonstrates both the validity and power of the C3H/10T1/2 system for Shh target gene discovery.

Age-related normograms of serum antimüllerian hormone levels in a population of infertile women: a multicenter study.

Objective: To produce age-related normograms for serum antimullerian hormone (AMH) level in infertile women without polycystic ovaries (non-PCO).Design: Retrospective cohort analysis.Setting: Fifteen academic reproductive centers.Patient(s): A total of 3,871 infertile women.Intervention(s): Blood sampling for AMH level.Main Outcome Measure(s): Serum AMH levels and correlation between age and different percentiles of AMH.Result(s): Age-related normograms for the 3rd, 10th, 25th, 50th, 75th, 90th, and 97th percentiles of AMH were produced. We found that the curves of AMH by age for the 3rd to 50th percentiles fit the model and appearance of linear relation, whereas the curves of >75th percentiles fit cubic relation. There were significant differences in AMH and FSH levels and in antral follicle count (AFC) among women aged 24-33 years, 34-38 years, and >= 39 years. Multivariate stepwise linear regression analysis of FSH, age, AFC, and the type of AMH kit as predictors of AMH level shows that all variables are independently associated with AMH level, in the following order: AFC, FSH, type of AMH kit, and age.Conclusion(s): Age-related normograms in non-PCO infertile women for the 3rd to 97th percentiles were produced. These normograms could provide a reference guide for the clinician to consult women with infertility. However, future validation with longitudinal data is still needed. (Fertil Steril (R) 2011; 95: 2359-63. (C) 2011 by American Society for Reproductive Medicine.)

Origins and functional evolution of Y chromosomes across mammals.

Y chromosomes underlie sex determination in mammals, but their repeat-rich nature has hampered sequencing and associated evolutionary studies. Here we trace Y evolution across 15 representative mammals on the basis of high-throughput genome and transcriptome sequencing. We uncover three independent sex chromosome originations in mammals and birds (the outgroup). The original placental and marsupial (therian) Y, containing the sex-determining gene SRY, emerged in the therian ancestor approximately 180 million years ago, in parallel with the first of five monotreme Y chromosomes, carrying the probable sex-determining gene AMH. The avian W chromosome arose approximately 140 million years ago in the bird ancestor. The small Y/W gene repertoires, enriched in regulatory functions, were rapidly defined following stratification (recombination arrest) and erosion events and have remained considerably stable. Despite expression decreases in therians, Y/W genes show notable conservation of proto-sex chromosome expression patterns, although various Y genes evolved testis-specificities through differential regulatory decay. Thus, although some genes evolved novel functions through spatial/temporal expression shifts, most Y genes probably endured, at least initially, because of dosage constraints.

Undetectable serum anti-Müllerian hormone levels and occurrence of ongoing pregnancy.

OBJECTIVE: To discuss, on the basis of the experience of two clinical cases and extensive literature review, the significance of extremely low levels of anti-Müllerian hormone (AMH), also known as Müllerian-inhibiting substance, in infertile women. DESIGN: Case report. SETTING: University-based infertility clinic at a medical center in Switzerland. PATIENT(S): Two women, 29 and 41 years of age and with a 2- and 4-year history of secondary infertility, respectively. INTERVENTION(S): Clinical, radiological, and biological investigation of infertility, including repeated measurements of the serum AMH with serial ELISA assays. MAIN OUTCOME MEASURE(S): Levels of AMH and development of ongoing pregnancy. RESULT(S): Both women had a spontaneous ongoing pregnancy despite undetectable AMH levels. CONCLUSION(S): Although it is helpful for day-to-day management of infertile patients, the predictive value of AMH for the occurrence of a spontaneous ongoing pregnancy has limits.

Serum antimüllerian hormone levels remain stable through the menstrual cycle and after oral or vaginal administration of synthetic sex steroids

Rapport de synthèse : Objectif de l'étude : étudier si l'administration orale ou vaginale d'hormones contraceptives influence les concentrations sériques d'hormone antimüllérienne (AMH). Design : essai prospectif chez des femmes recrutées par annonce. Les femmes désireuses d'avoir une contraception ont été randomisées entre une contraception orale et une contraception vaginale. Celles qui ne souhaitaient pas de contraception ont été incluses dans le groupe témoin. Cadre de l'étude : unité de médecine de la reproduction d'un hôpital universitaire. Patientes : vingt-quatre jeunes femmes en bonne santé avec des cycles menstruels réguliers qui n'avaient pas utilisé de contraception hormonale pendant les trois mois précédant l'étude. Intervention : contraception orale ou vaginale du 5ème au 25ème jour du cycle menstruel dans les groupes contraception versus pas de contraception dans le groupe témoin. Mesure d'issue : variations inter et intra-cycle des concentrations sériques d'AMH dans les trois groupes: groupe témoin en cycle spontané et groupes sous contraception oestroprogestative orale ou vaginale. Résultats : les fluctuations d'AMH observées pendant le cycle menstruel (variations intra-cycle) restent dans les valeurs des variations entre deux cycles (variations inter-cycles) tant chez les femmes en cycle spontané que chez les femmes sous contraception orale ou vaginale. Conclusions : nos résultats confirment que les concentrations sériques d'AMH restent stables pendant le cycle menstruel et indiquent qu'elles ne sont pas influencées par l'administration exogène de stéroïdes sexuels contraceptifs, que ce soit par voie orale ou vaginale.

Clinical uses of anti-Müllerian hormone assays : pitfalls and promises

Rapport de synthèse : OBJECTIF : étudier si les fluctuations sériques du taux d'hormone anti-müllérienne (AMH) sont liées à des différences méthodologiques entre les deux tests ELISA commercialement disponibles le kit de Beckman Coulter Immunotech (Fullerton, CA) et le kit de Diagnostic Systems Laboratories (Webster, TX). DESIGN : étude prospective au sein de deux services universitaires de médecine de la reproduction. POPULATION ETUDIEE : cent soixante-huit échantillons sanguins provenant de trois populations différentes ainsi que des dosages itératifs au cours du même cycle menstruel dans une population de dix volontaires. INTERVENTIONS : doubles mesures en aveugle des taux sériques d'AMH dans les 168 sera avec les deux kits commercialement disponibles. Sept dosages itératifs au cours du même cycle menstruel chez 10 volontaires normo-ovulantes. ANALYSE STATISTIQUE : régression linéaire pour l'étude de la corrélation entre les deux méthodes de dosage. Analyse des variances pour les dosages sériés en cours de cycle menstruel. RÉSULTATS : nous avons démontré une relation linéaire entre les deux méthodes, avec un coefficient de corrélation de 0,88 ainsi qu'une diminution faible mais statistiquement significative et concomitante avec l'ovulation du taux d'AMH sérique. CONCLUSION : les variations en cours de cycle menstruel rapportées par certains auteurs ne sont pas liées à des problèmes méthodologiques. La chute du taux d'AMH lors de l'ovulation est faible mais statistiquement significative. Les variations observées en cours de cycle restent toutefois inférieures aux variations intercycles et ne représentent de ce fait pas un obstacle au dosage de l'AMH à n'importe quel jour du cycle menstruel.

Statistically significant changes of antimüllerian hormone and inhibin levels during the physiologic menstrual cycle in reproductive age women.

OBJECTIVE: To define the dynamics of antimüllerian hormone (AMH) and inhibins during the physiologic menstrual cycle. DESIGN: Longitudinal study. SETTING: University hospital. PATIENT(S): 36 young, healthy, normal weight Caucasian women without medication. INTERVENTION(S): Normal ovulatory menstrual cycles were evaluated by regular blood sampling taken every other day and periovulatory every day. MAIN OUTCOME MEASURE(S): Serum concentrations of AMH, inhibin A and B, follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin, estradiol, progesterone, and free testosterone were measured in all blood samples. RESULT(S): Median AMH levels are statistically significantly higher in the late follicular compared with ovulation or the early luteal phase. There are statistically significant correlations between both AMH and FSH, and AMH and free testosterone in all cycle phases. Inhibin A increases strongly in the late follicular phase and peaks at day LH + 4. Inhibin B shows a broad midfollicular and a sharp early luteal peak, the difference being statistically significant between day LH + 4 and the earlier time points and between day LH + 2 and day LH. Although there is a negative association between inhibin A or B and the body mass index (BMI), there is no correlation between AMH and the BMI. CONCLUSION(S): Levels of AMH show a statistically significant change during the menstrual cycle and may influence the circulating gonadotropin and steroid hormone levels.

Entonación de las interrogativas absolutas del español meridional en habla espontánea

Este trabajo tiene como objetivos, en primer lugar, comprobar si los cuatro patrones interrogativos descritos para el castellano peninsular en pruebas de percepción se encuentran en el habla espontánea de los hablantes nativos de español peninsular meridional de Andalucía, Extremadura, Murcia, Castilla-La Mancha y Canarias; en segundo, saber si existen rasgos melódicos específicos de cada zona, y, finalmente, identificar los significados semántico-pragmáticos de cada patrón. Para llevar a cabo la investigación nos hemos basado en 186 preguntas absolutas, emitidas en un contexto de habla espontánea y siguiendo la metodología Análisis Melódico del Habla (AMH). Tras el análisis acústico, los resultados obtenidos han permitido: a) constatar la existencia de los cuatro patrones en las cinco zonas dialectales; b) determinar que, desde el punto de vista de la entonación lingüística, no parece que haya rasgos melódicos que distingan unas zonas de otras, sino una distinta representatividad de los patrones en cada una; y c) describir los significados sociopragmáticos apuntados por Escandell (1998, 2002), que nos permiten discriminar el patrón que se usa en cada contexto.

Sex Reversal in Zebrafish fancl Mutants is Caused by Tp53-Mediated Germ Cell Apoptosis

The molecular genetic mechanisms of sex determination are not known for most vertebrates, including zebrafish. We identified a mutation in the zebrafish fancl gene that causes homozygous mutants to develop as fertile males due to female-to-male sex reversal. Fancl is a member of the Fanconi Anemia/BRCA DNA repair pathway. Experiments showed that zebrafish fancl was expressed in developing germ cells in bipotential gonads at the critical time of sexual fate determination. Caspase-3 immunoassays revealed increased germ cell apoptosis in fancl mutants that compromised oocyte survival. In the absence of oocytes surviving through meiosis, somatic cells of mutant gonads did not maintain expression of the ovary gene cyp19a1a and did not down-regulate expression of the early testis gene amh; consequently, gonads masculinized and became testes. Remarkably, results showed that the introduction of a tp53 (p53) mutation into fancl mutants rescued the sex-reversal phenotype by reducing germ cell apoptosis and, thus, allowed fancl mutants to become fertile females. Our results show that Fancl function is not essential for spermatogonia and oogonia to become sperm or mature oocytes, but instead suggest that Fancl function is involved in the survival of developing oocytes through meiosis. This work reveals that Tp53-mediated germ cell apoptosis induces sex reversal after the mutation of a DNA-repair pathway gene by compromising the survival of oocytes and suggests the existence of an oocyte-derived signal that biases gonad fate towards the female developmental pathway and thereby controls zebrafish sex determination.

Estudo espectro eletroquímico de sistemas estruturados argila-corante

O comportamento eletroquímico e espectroeletroquímico do sistema montmorilonita-azul de metileno (MM-AM)/AMsolução foi investigado empregando-se a técnica de voltametria cíclica e de transiente, I x t (corrente versus tempo) sobre eletrodo de vidro ITO modificado pela deposição mecânica de filmes do compósito MM-AM. Os filmes foram preparados empregando três procedimentos: 1- na ausência de CTACl (surfactante, cloreto de cetiltrimetilamônio); 2- com incorporação do CTACl à região interlamelar; e 3- na presença do CTACl, porém, não incorporado à região interlamelar. A técnica espectrofotométrica foi empregada para a caracterização e para o estudo eletrocrômico dos filmes formados. Os resultados experimentais permitiram observar que: i- os filmes preparados com a incorporação prévia do AM a região interlamelar fornecem uma resposta eletroquímica mais eficiente com o aparecimento de um par de picos bem definido; ii- a adição do CTACl, de forma geral permite a obtenção de filmes mecanicamente mais estáveis; iii- com a inclusão prévia do CTACl na região interlamelar não ocorre o aparecimento da forma protonada do AM (AMH+); iv- na ausência da forma protonada o compósito não apresenta comportamento eletrocrômico.

Evidência da resposta folicular ao FSH pelo hormônio antimülleriano em mulheres ovulatórias

OBJETIVO: examinar a hipótese de que o nível sérico do hormônio antimülleriano (AMH) reflete a resposta dos folículos antrais à administração do FSH. MÉTODOS: estudo prospectivo, no qual foram incluídas 116 pacientes normo-ovulatórias inférteis submetidas à hiperestimulação ovariana controlada com agonista de GnRH e FSH. Depois de atingir a supressão pituitária e antes da administração de FSH (dia basal), o nível sérico de AMH foi mensurado. O número de folículos antrais foi determinado pela ultrassonografia no dia basal (folículos antrais precoces; 2 a 8 mm) e no dia da administração do hCG (dhCG; folículos pré-ovulatórios; >16 mm). A resposta folicular ao FSH foi determinada pela percentagem de folículos antrais precoces que atingiram os estágios pré-ovulatórios em resposta ao FSH (taxa de maturação). Foram estudadas as correlações do AMH com a idade das pacientes, número total de folículos antrais precoces e pré-ovulatórios, oócitos coletados, dose total de FSH na estimulação ovariana controlada e a taxa de maturação folicular. Para análise estatística, foram usados a regressão simples e o teste de Spearman's, com nível de significância de 5%. RESULTADOS: o nível sérico de AMH foi positivamente correlacionado com o número de folículos antrais precoces no dia basal (r=0,64; p<0,0001) e folículos pré-ovulatórios em dhCG (r=0,23; p=0,01). Excepcionalmente, o nível sérico de AMH foi negativamente correlacionado com a taxa de maturação (r=-0,24; p<0,008). CONCLUSÕES: o AMH atenua o desenvolvimento folicular em resposta à administração do FSH.

Hormônio anti-Mülleriano sérico para predição da resposta ovariana em ciclos de reprodução assistida

OBJETIVOS: Comparar as concentrações séricas do hormônio anti- Mülleriano (AMH) no sétimo dia de estimulação ovariana em pacientes boas e más respondedoras. MÉTODOS: Foram incluídas 19 mulheres com idade ≥35 anos, ciclos menstruais regulares e que foram submetidas à estimulação ovariana para reprodução assistida. Foram excluídas mulheres portadoras de endometriose ou síndrome dos ovários policísticos ou aquelas submetidas previamente à cirurgia ovariana. Foram coletadas amostras de sangue periférico no dia basal e no sétimo dia de estimulação para dosagens de AMH, hormônio folículo estimulante (FSH) e estradiol. Os níveis de AMH foram avaliados pelo método de enzimoimunoensaio (ELISA) e os níveis de FSH e estradiol foram avaliados por imunoquimioluminescência. Ao final do ciclo, as pacientes foram classificadas como normo (obtenção de quatro ou mais oócitos durante a captação) ou más respondedoras (obtenção de menos de quatro oócitos ou cancelamento do ciclo por má resposta) e analisadas comparativamente em relação aos níveis hormonais, duração da estimulação ovariana, número de folículos recrutados, embriões produzidos e transferidos através do teste t. A associação entre os níveis de AMH e os parâmetros acima também foi avaliada pelo teste de correlação de Spearman. RESULTADOS: Não houve diferença significativa entre os grupos para os níveis de AMH, FSH e estradiol no dia basal e no sétimo dia de estimulação ovariana, sendo observada correlação significativa entre os níveis de AMH do sétimo dia e a quantidade total de FSH exógeno utilizada (p=0,02). CONCLUSÕES: Os níveis de AMH obtidos no sétimo dia do ciclo de estimulação ovariana não parecem predizer o padrão de resposta ovariana, não sendo recomendadas as suas dosagens para esta finalidade.