LH (Trp8Arg/Ile15Thr), LHR (insLQ) and FSHR (Asn680Ser) polymorphisms genotypic prevalence in women with endometriosis and infertility.

PURPOSE: To verify if polymorphisms of LH (Trp8Arg/Ile15Thr), LH receptor (insLQ), and FSH receptor (Asn680Ser) are associated with endometriosis and infertility. METHODS: This is a prospective case-control study. Sixty-seven patients with endometriosis and infertility (study group) and 65 healthy fertile patients (control group) were enrolled in the study between July 2010 and July 2013. All patients had their endometriosis diagnosis made or excluded by laparoscopic surgery; study group was submitted to the surgery for infertility investigation and control group for tubal ligation. Day-3 serum hormones were collected from all patients. Analysis of nucleotide mutations for LH polymorphisms (Trp8Arg and Ile15Thr), LHR polymorphism (insLQ), and FSHR polymorphism (Asn680Ser) were performed by PCR. RESULTS: Day-3 FSH, estradiol and LH serum levels were not different between the groups, while CA-125 was higher in patients with endometriosis and infertility. All polymorphisms studied were in Hardy-Weinberg equilibrium. The prevalence of insLQ was significantly higher in patients with endometriosis and infertility (P = 0.005). Allele occurrence in control group was 0.10 versus 0.25 in infertile endometriosis group (P = 0.001). There was no difference regarding Trp8Arg/Ile15Thr (P > 0.05) and Asn680Ser (P > 0.05) prevalence between groups. CONCLUSION: This is the first time that prevalence of insLQ was shown to be higher in patients with endometriosis and infertility than in healthy fertile patients. There was no difference in LH and FSHR polymorphisms' prevalence between groups.

Gut inflammation and expression of ICC in a fetal lamb model of fetoscopic intervention for gastroschisis.

BACKGROUND: The pathogenesis of intestinal dysmotility in gastroschisis is not completely understood. Peel formation and disorganization of interstitial Cajal cells (ICC) have been proposed in humans. The aim of this study was to evaluate the impact of prenatal coverage of gastroschisis on gut inflammation and expression of ICC in a fetal lamb model. METHODS: Twenty-one German blackhead sheep with an abdominal wall defect that was created fetoscopically on day 77 of 145 days gestation were used in this study. Intrauterine surgery with the aim to cover the defect was performed 3 weeks later; two fetuses were covered completely, 5 partially and 11 remained uncovered. Three fetuses without gastroschisis were used as controls. All fetuses were retrieved by cesarean section at day 135. Samples of the small intestine were stained with hematoxylin and eosin for histologic analysis of peel formation and serosal and muscular thickness. For ICC detection, immunohistochemistry using anti-CD117 (c-Kit) antibody was used. RESULTS: In all samples with exposure to amniotic fluid, peel formation and significantly decreased ICC were found. Complete coverage reduced peel formation and disorganization of ICC compared to uncovered animals almost to the level of controls. CONCLUSIONS: Peel formation and ICC derangement were significantly reduced by prenatal coverage of gastroschisis. Moreover, this animal model mimics the histopathological bowel changes as seen in human gastroschisis and may, therefore, be used for further research on the pathophysiology and fetal therapy of this malformation.

Abdominal wall incision with or without exteriorization of bowel: results from a fetal lamb model for the embryogenesis of gastroschisis.

INTRODUCTION: The embryogenesis of gastroschisis is not completely understood. The aim of our study was to evaluate the impact of a simple abdominal wall defect versus a defect including eviscerated intestine or omentum for the development of gastroschisis in a fetal lamb model. MATERIAL AND METHODS: At mid-gestation (day 77) an abdominal wall defect was fetoscopically created with three different approaches in 19 German blackhead sheep. The intestine was eviscerated in 7 fetuses (group 1). The peritoneum was incised and a patch of the omentum pulled through the incision in 5 fetuses (group 2). In 7 fetuses (group 3) the skin and rectus muscle were incised until the peritoneum was visible. In this group, no abdominal contents were exteriorized and the peritoneum was left intact. A second fetoscopic procedure was performed 21 days later, assessing the condition and extension of eviscerated bowel. The fetus was retrieved by Cesarean section on day 132 and evaluated. RESULTS: The second fetoscopy acting as a control for the creation of gastroschisis demonstrated eviscerated and inflamed intestine in all 3 groups. The amount of eviscerated intestine did not appear to depend on the size of the defect nor on its duration. DISCUSSION: It appears that a simple incision of the abdominal wall with intact peritoneum is sufficient for the development of gastroschisis in a fetal sheep model. This finding may improve the understanding of the etiology of gastroschisis.

Fetoscopic management of gastroschisis in a lamb model.

BACKGROUND: Gastroschisis is a malformation consisting of an abdominal wall defect with eviscerated bowel. Its standard treatment is postnatal repositioning or temporary prosthetic bag placement. The aim of our study is to evaluate the feasibility of its fetoscopic management in a lamb model. METHODS: At mid-gestation (day 75), gastroschisis was created in eight German blackhead sheep. A second fetoscopic procedure was performed on day 105, assessing the viability, extension, and potential for repositioning of eviscerated bowel. The fetus was retrieved by Cesarean section on day 132 and evaluated. RESULTS: In six fetuses gastroschisis could be successfully created and assessed with fetoscopy. Two fetuses were lost due to technical complications. Primary repair by repositioning the intestine into the abdomen was not possible because it grew into an inflammatory conglomerate too large for the small fetal abdominal cavity. CONCLUSIONS: Although technically demanding, we were able to produce and reassess six cases of gastroschisis by fetoscopy. As primary repositioning appears unfavorable, fetoscopic prosthetic bag placement may become an alternative.

Does the degree of hypothalamic-pituitary-ovarian recovery after oral contraceptive pills affect outcomes of IVF/ICSI cycles receiving GnRH-antagonist adjuvant therapy in women over 35 years of age?

PURPOSE: To evaluate if the degree of recovery of serum gonadotropins after oral contraceptive pills (OCP) pretreatment has an impact on ovarian response in GnRH-antagonist IVF cycles in women of advanced maternal age. METHODS: In this retrospective cohort study, we included 98 women 35-42 years undergoing their first IVF cycle receiving gonadotropins and a fixed GnRH-antagonist adjuvant protocol. Data analysis was carried out according to changes in serum FSH, LH and estradiol (E(2)) levels (basal and post-OCP) divided in quartiles, and also according to absolute levels. The main outcomes were peak serum E(2), number of mature oocytes retrieved, length of stimulation, and amount of gonadotropins used. RESULTS: By quartile analysis, patients with the highest levels of serum gonadotropins suppression and also patients with gonadotropin rebound needed larger amounts of LH during the treatment. On the other hand, women with absolute suppression of FSH/LH had increased length of stimulation. CONCLUSIONS: The results of this study provide data that assist in clinical management. Gonadotropin serum levels after OCP treatment provide information for optimization of supplementation with LH in GnRH-antagonist cycles in women over age 35.

Alterations in dendritic cell function in aged mice: potential implications for immunotherapy design.

It is known that immune system functions decrease with age, and that adaptive immune responses, especially CD4+ T cell function, seem to be the main affected point in immunity with aging. Dendritic cells (DC) are the major antigen presenting cell (APC), and at least part of the defects observed in adaptive immunity of aged individuals could be due to diminished potential of bone marrow to generate new DC, or defects in DC function. In this study, we investigated if the ability of aged bone marrow (BM) to generate new DC in vitro, as well as aged BM-derived DC responses to lypopolysaccharide (LPS). Because DC are important tools in newly developing anti-tumor therapies, we also studied the ability of aged DC to phagocytose and present antigen from necrotic tumor cells. We found that aged BM generated fewer DC in vitro compared to young BM. While LPS-induced DC maturation is reduced in DC of aged mice, a high TNF-alpha production is observed in aged DC even without LPS stimulation. While phagocytosis of tumor cells is not affected by age, and DC derived from aged BM show a higher TNF-alpha production in response to phagocytosis, presentation of tumor antigens was decreased in aged DC. Because class II upregulation in response to phagocytosis was similar between aged and young DC, this could indicate an age associated processing defect in the exogenous pathway. These findings suggest that age of BM used to generate DC does not impair their phagocytic ability or TNF-alpha production, however leads to a decreased yield in mature DC, reduced response to LPS, and diminished antigen processing/presentation potential. Our results are relevant to optimization DC-based vaccine design for aged populations.

Alterations in expression of endometrial milk fat globule-EGF factor 8 (MFG-E8) and leukemia inhibitory factor (LIF) in patients with infertility and endometriosis.

OBJECTIVE: The aim of this study was to compare the endometrial expression of milk fat globule-EGF factor 8 (MFG-E8), its receptor integrin αvß3, and leukemia inhibitory factor (LIF) in patients with endometriosis and infertility and in healthy fertile patients during the window of implantation. METHODS: Five patients with peritoneal endometriosis and infertility (case group) and four healthy fertile patients (control group) were recruited. All patients were either diagnosed with or ruled out for endometriosis by laparoscopic surgery; the case group underwent surgery for infertility investigation and the control group for tubal ligation. Endometrial biopsies were performed in all patients during the window of implantation (LH+8 to LH+10), and then the samples were analyzed by immunochemistry for MFG-E8, integrin αvß3, and LIF. RESULTS: In patients with endometriosis and infertility, expression of MFG-E8 was significantly increased in the glandular epithelium when compared to healthy fertile patients (p<0.001). Moreover, LIF expression was lower in patients with endometriosis and infertility (p<0.05). Nevertheless, we found no difference in integrin αvß3 expression between the groups (p=0.084). CONCLUSION: This study showed for the first time that MFG-E8 expression is impaired in the endometrium of patients with endometriosis and infertility during the window of implantation. Moreover, LIF is also diminished in the endometrium of these patients as shown before.

Role for the endometrial epithelial protein MFG-E8 and its receptor integrin αvß3 in human implantation: results of an in vitro trophoblast attachment study using established human cell lines.

OBJECTIVE: To investigate the role of MFG-E8 and its receptor integrin αvß3 in the attachment of trophoblast cells to the endometrial epithelium. DESIGN: Experimental in vitro study. SETTING: Academic center. PATIENT(S): None. INTERVENTION(S): By using a well-differentiated endometrial adenocarcinoma cell line (Ishikawa cells) and choriocarcinoma human trophoblast cells (Jar cells), an in vitro assay mimicking human implantation was established. To investigate the impact of blocking MFG-E8 and integrin αvß3, we pretreated the cell lines with antibodies against those proteins at different concentrations before the attachment assay. MAIN OUTCOME MEASURE(S): Attachment rate of Jar spheroids to the epithelial cell monolayer. RESULT(S): Pretreatment of Ishikawa cells with anti-MFG-E8 antibody caused a dose-dependent and significant inhibition of attachment. On the other hand, pretreatment of Jar spheroids did not result in a significant effect on the attachment rate. Pretreatment of Ishikawa cells as well as Jar spheroids with anti-integrin αvß3 antibodies resulted in a dose-dependent, significant inhibition of attachment. CONCLUSION(S): This study showed that blocking MFG-E8 and its receptor integrin αvß3 in Ishikawa cells diminishes Jar spheroid attachment. Moreover, blocking integrin αvß3 in the trophoblastic cells also diminished their attachment to the Ishikawa monolayer.

Office hysteroscopy study in consecutive miscarriage patients.

OBJECTIVE: To assess the prevalence of uterine anatomical abnormalities found by office diagnostic hysteroscopy in a population of patients experiencing more than two consecutive miscarriages and compare the prevalence of uterine abnormalities between patients with two miscarriages and those with three or more consecutive miscarriages. METHODS: A cross-sectional study of 66 patients with two or more consecutive miscarriages diagnosis was conducted. Patients were divided into two groups: Group A (up to two miscarriages, 23 patients), and Group B (3 miscarriages, 43 patients). They underwent an outpatient diagnostic hysteroscopy study, with either congenital or acquired abnormalities of the uterine cavity being identified. RESULTS: Uterine changes were found in 22 (33.3%) patients, with 9 cases of congenital changes [arcuate uterus (4 cases), septate uterus (2 cases), and bicornuate uterus (1 case)], and 13 patients with acquired changes [intrauterine adhesions (7 cases), endometrial polyp (4 cases), and uterine leiomyoma (2 cases)]. No significant differences were found between the groups as regarding both acquired and congenital uterine changes. A positive correlation was found between anatomical changes on hysteroscopy and number of miscarriages (r = 0.31; p = 0.02). CONCLUSION: Patients with more than two miscarriages have a high prevalence of uterine cavity abnormalities diagnosed by hysteroscopy; however there are no differences in prevalence or distribution of these lesions related to the number of recurrent miscarriages.

Mycobacterium tuberculosis heat-shock protein 70 impairs maturation of dendritic cells from bone marrow precursors, induces interleukin-10 production and inhibits T-cell proliferation in vitro.

In different inflammatory disease models, heat-shock proteins (hsp) and hsp-derived peptides have been demonstrated to possess anti-inflammatory properties. While some studies have shown that hsp can directly interact with antigen-presenting cells, others report that bacterial hsp can induce specific T cells with regulatory phenotypes. Effective characterization of the immunomodulatory effects of hsp 70, however, has historically been confounded by lipopolysaccharide (LPS) contamination. In this study, we compared the effects of LPS-free Mycobacterial tuberculosis hsp 70 (TBhsp70) and its possible contaminants on dendritic cells (DC). We demonstrate herein that LPS-free TBhsp70 inhibits murine DC maturation in vitro, while LPS-contaminated TBhsp70 induces DC maturation. Mock recombinant preparations have no effect. In contrast to LPS, TBhsp70 does not induce tumour necrosis factor-alpha production by DC, but interleukin-10. In vivo, only LPS-contaminated TBhsp70 induces up-regulation of CD86 in splenic mature DC. Finally, TBhsp70 inhibited phytohaemagglutinin-induced T-cell proliferation. Our results support the hypothesis that TBhsp70 does not have inflammatory potential, but rather has immunosuppressive properties.

Histeroscopia ambulatorial em casos de abortamento consecutivo / Office hysteroscopy study in consecutive miscarriage patients

OBJETIVO: Avaliar a prevalência de alterações anatômicas uterinas diagnosticadas através da histeroscopia ambulatorial em uma população de pacientes com mais de dois abortamentos consecutivos. Comparar a prevalência de alterações uterinas entre as pacientes com dois abortos em relação as pacientes com três ou mais abortamentos de repetição. MÉTODOS: Foi realizado um estudo transversal em 66 pacientes com diagnóstico de dois ou mais abortamentos de repetição. As pacientes foram divididas em dois grupos: Grupo A (até dois abortamentos, 23 pacientes) e Grupo B (três ou mais abortamentos, 43 pacientes), sendo submetidas à histeroscopia diagnóstica ambulatorial em que foram identificadas alterações congênitas e adquiridas da cavidade uterina. RESULTADOS: Foram encontradas em 22 (33,3 por cento) pacientes alterações uterinas, sendo em nove casos alterações congênitas [útero arqueado (quatro casos), septo uterino (dois casos) e útero bicorno (um caso)] e em 13 pacientes alterações adquiridas [sinéquia (sete casos), pólipo endometrial (quatro casos) e mioma uterino (dois casos). Não houve diferença significativa entre grupos em relação às alterações uterinas adquiridas e congênitas. Foi encontrada uma correlação positiva entre alterações anatômicas na histeroscopia e número de abortamentos (r = 0,31; p = 0,02). CONCLUSÃO: As pacientes com mais de dois abortamentos apresentam uma alta prevalência de alterações uterinas diagnosticadas por histeroscopia. No entanto não há diferença na prevalência ou na distribuição das lesões em relação ao número de abortamentos.

OBJECTIVE: To assess the prevalence of uterine anatomical abnormalities found by office diagnostic hysteroscopy in a population of patients experiencing more than two consecutive miscarriages and compare the prevalence of uterine abnormalities between patients with two miscarriages and those with three or more consecutive miscarriages. METHODS: A cross-sectional study of 66 patients with two or more consecutive miscarriages diagnosis was conducted. Patients were divided into two groups: Group A (up to two miscarriages, 23 patients), and Group B (3 miscarriages, 43 patients). They underwent an outpatient diagnostic hysteroscopy study, with either congenital or acquired abnormalities of the uterine cavity being identified. RESULTS: Uterine changes were found in 22 (33.3 percent) patients, with 9 cases of congenital changes [arcuate uterus (4 cases), septate uterus (2 cases), and bicornuate uterus (1 case)], and 13 patients with acquired changes [intrauterine adhesions (7 cases), endometrial polyp (4 cases), and uterine leiomyoma (2 cases)]. No significant differences were found between the groups as regarding both acquired and congenital uterine changes. A positive correlation was found between anatomical changes on hysteroscopy and number of miscarriages (r = 0.31; p = 0.02). CONCLUSION: Patients with more than two miscarriages have a high prevalence of uterine cavity abnormalities diagnosed by hysteroscopy; however there are no differences in prevalence or distribution of these lesions related to the number of recurrent miscarriages.

Endometriose e infertilidade / Endometriosis and infertility

Os autores fazem uma revisão bibliográfica sobre endometriose associada à infertilidade com enfoque aos aspectos terapêuticos da doença.

Investigação de amenorréia / Inquiry of amenorrhea

Amenorréia é a ausência ou parada anormal de sangramento menstrual na menacme; para conduzir sua investigação, é importante descartar causas fisiológicas (gravidez e puerpério). além disso, deve-se conhecer as diversas situações patológicas que se manifestam como amenorréia, as quais podem ocorrer por disfunção do eixo hipotálamo-hipófise-gonadal o a anatomia do trato genital. nesse artigo serão revisadas as principais etiologias da amenorréia e sua investigação.