Category Archives: Human Reproduction

BACKGROUND

Endometriosis is a multifactorial gynecological disease characterized by the presence of functional endometrium-like tissue in ectopic sites. Several studies have focused on elucidating the immunological, endocrine, environmental and genetic factors involved in endometriosis. However, its pathogenesis is still unclear.

METHODS

High-resolution comparative genomic hybridization was applied to screen for genomic imbalances in laser microdissected stromal and epithelial cells from 20 endometriotic lesions and three samples of eutopic endometrium derived from eight patients. The expression of seven stemness-related markers (CD9, CD13, CD24, CD34, CD133, CD117/c-Kit and Oct-4) in endometrial tissue samples was evaluated by immunohistochemistry.

RESULTS

Samples of eutopic endometrium showed normal genomic profiles. In ectopic tissues, an average of 68 genomic imbalances was detected per sample. DNA losses were more frequently detected and involved mainly 3p, 5q, 7p, 9p, 11q, 16q, 18q and 19q. Many of the genomic imbalances detected were common to endometriotic stroma and epithelia and also among different endometriotic sites from the same patient. These findings suggested a clonal origin of the endometriotic cells and the putative involvement of stem cells. Positive immunostaining for CD9, CD34, c-Kit and Oct-4 markers was detected in isolated epithelial and/or stromal cells in eutopic and ectopic endometrium in the majority of cases.

CONCLUSIONS

The presence of shared genomic alterations in stromal and epithelial cells from different anatomical sites of the same patient and the expression of stemness-related markers suggested that endometriosis arises as a clonal proliferation with the putative involvement of stem cells.

Source:
http://humrep.oxfordjournals.org/cgi/content/short/27/11/3187?rss=1

STUDY QUESTION

How is the expression of nectins and nectin-like molecules (Necls) detected by immunostaining altered by endometriosis?

SUMMARY ANSWER

Our results suggest that Nectin-1, -3, -4 and Necl-2 may contribute to the pathogenesis of endometriosis. Immunostaining of nectins and Necls varies according to the anatomical location of endometriosis.

WHAT IS KNOWN AND WHAT THIS PAPER ADDS

Nectin and Necl molecules are immunoglobulin-like cell adhesion molecules involved in apoptosis, cell proliferation and in metastases. Previous studies have demonstrated the involvement of adhesion molecules in the development of endometriotic lesions but no data exist on immunostaining of nectins and Necls molecules in endometriosis.

DESIGN, PARTICIPANTS AND SETTING

This retrospective study was conducted in a tertiary-care hospital (Tenon Hospital, Paris, France). Samples were collected from 55 women undergoing endometrial biopsy or surgery for endometriosis and 20 controls having hysterectomy or endometrial biopsy for other reasons; multiple samples were collected from 15 women. We studied the immunostaining of Nectin-1, -3, -4 and Necl-2 in secretory and proliferative endometrium from women with (n = 20) or without endometriosis (i.e. control group, n = 20), and in peritoneal (n = 20), ovarian (n = 20) and colorectal endometriosis (n = 20).

MAIN RESULTS

Semi-quantitative immunostaining demonstrated that (1) Necl-2 staining was stronger in all types of endometriotic lesions than in the eutopic endometrium from patients with endometriosis (P < 0.0125) and in ovarian endometriotic cysts compared with other locations (P < 0.001); (2) Nectin-3 staining was stronger in the eutopic endometrium of patients with endometriosis compared with controls (P = 0.03) and in all endometriotic lesions compared with the eutopic endometrium from patients with endometriosis (P < 0.0125); (3) Nectin-4, staining was stronger in the eutopic endometrium of patients with endometriosis compared with controls (P = 0.04) and (4) Nectin-1 staining was significantly increased in colorectal endometriosis compared with other locations (P = 0.004).

BIAS, CONFOUNDING AND OTHER REASONS FOR CAUTION

We did not assess the pattern of expression in endometriosis of all nectins and Necl molecules. Indeed, Necl-5 is implicated in many pathophysiological processes such as cell movement and proliferation with potential relevance to endometriosis.

GENERALISABILITY TO OTHER POPULATIONS

At present, few data on implication of nectins and Necl molecules in endometriosis exist. Hence, our results should be confirmed by further quantitative studies at protein or RNA levels.

STUDY FUNDING/COMPETING INTEREST(S)

No funding source. All the authors declare no conflict of interest.

Source:
http://humrep.oxfordjournals.org/cgi/content/short/27/11/3179?rss=1

STUDY QUESTION

How patient-centered are two included specialized endometriosis clinics relative to each other and how can they improve the patient-centeredness of their care?

SUMMARY ANSWER

The validated ENDOCARE questionnaire (ECQ) reliably concluded that the adjusted overall patient-centeredness did not differ between the clinics, that each clinic was significantly more patient-centered for 2 out of 10 dimensions of patient-centered endometriosis care and that clinics 1 and 2 had to improve 8 and 13 specific care aspects, respectively.

WHAT IS KNOWN ALREADY

Patient-centered endometriosis care is essential to high-quality care and is defined by 10 dimensions. The ECQ was developed, validated and proved to be reliable in a European setting of self-reported endometriosis patients but had not yet been used at a clinic level for quality management.

STUDY DESIGN, SIZE, DURATION

A cross-sectional survey was disseminated in 2011 to all 514 women diagnosed with endometriosis during a laparoscopy indicated for pain and/or infertility during a retrospective 2-year period (2009–2010) in two university clinics from two different European countries.

PARTICIPANTS/MATERIALS, SETTING, METHODS

In total 337 patients completed the ECQ (216 and 121 per clinic). Respondents had a mean age of 34.3 years. Three in four reported a surgical diagnosis of moderate or severe endometriosis and the majority reported surgical treatment by a multidisciplinary team. The ECQ assessed the 10 dimensions of patient-centeredness, more specifically whether the health-care performance, as perceived by patients, measured up to what is important to patients in general.

MAIN RESULTS

The ECQ was completed by 337 respondents (response rate = 65.6%). Reliability and validity of the ECQ for use on clinic level were confirmed. Clinics did not differ in overall mean importance scores; importance rankings of the ECQ dimensions were almost identical. The overall patient-centeredness scores (PCS), adjusted for education level, did not discriminate between the clinics. However, the adjusted PCS for the dimensions ‘clinic staff’ and ‘technical skills’ were significantly better in clinic 1, whereas the dimensions ‘physical comfort’ and ‘access to care’ were significantly better in clinic 2. There were 8 (clinic 1) and 13 (clinic 2) targets identified for joint and cross-clinic improvement.

LIMITATIONS, REASONS FOR CAUTION

Response rates were relatively high. Recall bias was the most important limitation and research in more clinics is needed to define the statistical discriminative value of the ECQ.

WIDER IMPLICATIONS OF THE FINDINGS

European endometriosis clinics can use the validated ECQ for reliable assessment of their ‘patient-centeredness’, for comparison with others and for setting specific targets to improve the patient-centeredness of their endometriosis care, to plan interventions, and to evaluate their effectiveness.

STUDY FUNDING/COMPETING INTEREST

This work was funded by KU Leuven and European Network of Endometriosis (ENE), supported by the European Commission (Public Health Executive Agency). No competing interests are declared.

Source:
http://humrep.oxfordjournals.org/cgi/content/short/27/11/3168?rss=1

STUDY QUESTION

Is decorin (DCN), a putative modulator of growth factor (GF) signaling, expressed in the primate ovary and does it play a role in ovarian biology?

SUMMARY ANSWER

DCN expression in the theca, the corpus luteum (CL), its presence in the follicular fluid (FF) and its actions revealed in human IVF-derived granulosa cells (GCs), suggest that it plays multiple roles in the ovary including folliculogenesis, ovulation and survival of the CL.

WHAT IS KNOWN ALREADY

DCN is a secreted proteoglycan, which has a structural role in the extracellular matrix (ECM) and also interferes with the signaling of multiple GF/GF receptors (GFRs). However, DCN expression and action in the primate ovary has yet to be determined.

STUDY DESIGN, SIZE, DURATION

Archival human and monkey ovarian samples were analyzed. Studies were conducted using FF and GC samples collected from IVF patients.

PARTICIPANTS/MATERIALS, SETTING, METHODS

Immunohistochemistry, western blotting, RT–PCR, quantitative RT–PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA) studies were complemented by cellular studies, including the measurements of intracellular Ca²⁺, reactive oxygen species (ROS), epidermal GF receptor (EGFR) phosphorylation by DCN and caspase activity.

MAIN RESULTS AND THE ROLE OF CHANCE

Immunohistochemistry revealed strong DCN staining in the connective tissue and follicular thecal compartments, but not in GCs of pre-antral and antral follicles. Pre-ovulatory follicles could not be studied, but DCN was associated with connective tissue of CL samples and the cytoplasm of luteal cells. DCN expression in monkey CL doubled (P < 0.05) towards the end of the luteal lifespan. DCN was found in human FF obtained from IVF patients (mean: 12.9 ng/ml; n = 20) as determined by ELISA. DCN mRNA and/or protein were detected in freshly isolated and cultured, luteinized human GCs. In the latter, exogenous human recombinant DCN increased intracellular Ca²⁺ levels and induced the production of ROS in a concentration-dependent manner. DCN, like epidermal GF, phosphorylated EGFR significantly (P < 0.05) and reduced the activity of caspase 3/7 in cultured GCs. The data indicate the expression of DCN in the theca of growing follicles, in FF of ovulatory follicles and in the CL. Therefore, DCN may exert paracrine actions via GF/GFR systems in multiple ovarian compartments.

LIMITATIONS, REASONS FOR CAUTION

Functional studies were performed in cultures of human luteinized GCs, which are an apt model but may not fully mirror the pre-ovulatory GC compartment or the CL. Other human ovarian cells, including the thecal cells, were not available.

WIDER IMPLICATIONS OF THE FINDINGS

In accordance with its evolving roles in other organs, ovarian DCN is an ECM-associated component, which acts as a multifunctional regulator of GF signaling in the primate ovary. DCN may thus be involved in folliculogenesis, ovulation and the regulation of the CL survival in primates.

STUDY FUNDING/COMPETING INTEREST(S)

This study was supported by Deutsche Forschungsgemeinschaft (DFG) MA1080/17-3 and in part DFG MA1080/21-1 (to AM), NIH grants HD24870 (S.R.O. and R.L.S.), the Eunice Kennedy Shriver NICHD/NIH through cooperative agreement HD18185 as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research (S.R.O.) and 8P51OD011092-53 for the operation of the Oregon National Primate Research Center (G.A.D., J.D.H., S.R.O. and R.L.S).

Source:
http://humrep.oxfordjournals.org/cgi/content/short/27/11/3249?rss=1

BACKGROUND

Mammalian spermatogenesis is a process that involves a complex expression program in both somatic and germ cells present in the male gonad. A number of studies have attempted to define the transcriptome of male meiosis and gametogenesis in rodents and primates. Few human transcripts, however, have been associated with testicular somatic cells and germ cells at different post-natal developmental stages and little is known about their level of germline-specificity compared with non-testicular tissues.

METHODS

We quantified human transcripts using GeneChips and a total of 47 biopsies from prepubertal children diagnosed with undescended testis, infertile adult patients whose spermatogenesis is arrested at consecutive stages and fertile control individuals. These results were integrated with data from enriched normal germ cells, non-testicular expression data, phenotype information, predicted regulatory DNA-binding motifs and interactome data.

RESULTS

Among 3580 genes for which we found differential transcript concentrations in somatic and germ cells present in human testis, 933 were undetectable in 45 embryonic and adult non-testicular tissues, including many that were corroborated at protein level by published gene annotation data and histological high-throughput protein immunodetection assays. Using motif enrichment analyses, we identified regulatory promoter elements likely involved in germline development. Finally, we constructed a regulatory disease network for human fertility by integrating expression signals, interactome information, phenotypes and functional annotation data.

CONCLUSIONS

Our results provide broad insight into the post-natal human testicular transcriptome at the level of cell populations and in a global somatic tissular context. Furthermore, they yield clues for genetic causes of male infertility and will facilitate the identification of novel cancer/testis genes as targets for cancer immunotherapies.

Source:
http://humrep.oxfordjournals.org/cgi/content/short/27/11/3233?rss=1

BACKGROUND

Little is known about the long-term course taken in life by couples who had undergone medically assisted reproduction (MAR). The aim of this study was to find out in a large sample whether, in comparison with parents, involuntarily childless couples have a different subjective perception of overall and specific quality of life over a period of >10 years.

METHODS

Between 1994 and 1997, 564 couples participated in the initial Heidelberg Fertility Consultation Service study of psychosocial aspects of infertility. In March 2008, a follow-up questionnaire was sent to all of these couples. Both partners were asked about the current status of their desire for a child and their satisfaction with life, their self-esteem, partnership, sexuality and career, as well as their current attitude towards the MAR they had undergone and experience of the process.

RESULTS

The final sample consisted of 148 couples and 60 women (response rate: 41% of the women and 31% of the men contacted). Fifty-nine percent of the women had at least one genetically related child, 11% had a foster or adopted child and 30% remained childless. Comparisons of psychological variables between parents and childless couples were done for the 148 couples only. Post-MAR parents indicated significantly higher self-esteem than childless couples (P < 0.01) and were more inclined to go through the infertility treatment again than childless couples (P < 0.001 for women, P < 0.05 for men). Positive aspects of infertility were seen more often by childless couples than by parents (P < 0.001). Childless women reported more occupational satisfaction than mothers (P < 0.01), while no such difference was identified in the male partners. Concerning overall life satisfaction, satisfaction with friendships and the partnership, and sexual satisfaction there were no statistically significant differences between childless women/men and mothers/fathers.

CONCLUSIONS

Overall, our 10-year follow-up survey indicated good psychological adjustment both in childless couples and in post-MAR parents. A decline of sexual satisfaction in childless couples (often reported in the literature) was not observed in this large sample. Quality of life in the long-term can safely be said to be high, both in the definitively childless couples and the post-MAR parents. These findings should be integrated into the information and counselling for would-be parents prior to infertility treatment. A major limitation of this study is that the majority of women and men from the initial study did not respond in our follow-up study.

Source:
http://humrep.oxfordjournals.org/cgi/content/short/27/11/3226?rss=1