Research In Reproductive Medicine Research Articles

Trends in clinical reproductive medicine research: 10 years of growth

To study the most important metrics of publication in the field of reproductive medicine over the decade 2003-2012 to aid in discerning the clinical, social, and epidemiologic implications of this relatively new but rapidly emerging area in medical sciences. Bibliometric analysis of most-cited publications from Web of Science databases. Not applicable. None. None. Most productive and frequently cited investigators, institutions, and countries and specific areas of research, scientific collaborations, and comparison of the growth of reproductive medicine research compared with other areas of medical investigation such as obstetrics and gynecology and related science categories. We found that 90 investigators with more than 1,000 citations had jointly published 4,010 articles. A continued rise in the impact factor of reproductive medicine journals was seen. The number of publications in reproductive medicine grew more rapidly compared with other science categories. Presently 22% of highly cited articles in reproductive medicine research are published in journals belonging to science categories outside reproductive medicine. The most-cited study groups are situated in the Netherlands, Belgium, Spain, the United States, and the United Kingdom, and collaborative studies have been increasing. Reproductive medicine research and subsequent clinical development have attained scientific growth and maturity. High-quality research is increasingly being published in high-impact journals. The increase in (inter)national collaborations seems to be key to the field's success.

Current Status and Application of Gene Trapping

With the completion of genome sequencing on more and more organisms,post-genomic era has begun. Gene trapping,as a powerful tool in the functional-genomic era,has been widespreadly applicated in many fields of biomedicine research because of its great merits in cloning,finding novel genes and unraveling their biological functions. The principles,classifications,manipulation procedures,advantages and disadvantages of gene trap technique and its application in developmental biology,novel gene identification, stem cells differentiation,cancer and reproductive medicine research are discussed,which will provide helpful references for relevant researchers.

How are neonatal and maternal outcomes reported in randomised controlled trials (RCTs) in reproductive medicine?

How do randomised controlled trials (RCTs) in reproductive medicine report maternal and neonatal outcomes, specifically singleton live birth? Despite the widespread appeal to use singleton live birth as the outcome measure in subfertility trials, 80% of RCTs fail to do so, and fail to report on neonatal and maternal outcomes. The aim of reproductive medicine is to assist subfertile couples in their wish to have children. A decade ago it was proposed to use singleton live birth as the outcome measure. We assessed whether clinical research has followed this recommendation, and how neonatal/maternal outcomes are reported. A review of the published literature from 1 January 1966 to 31 December 2012 was performed using the Cochrane database. We compared the time periods before and after 2004; the year after ESHRE recommended the use of singleton live birth. We searched the Cochrane database for RCTs in reproductive medicine, and recorded the number of studies that used singleton live birth as the outcome measure. We also recorded the reporting neonatal and maternal outcomes. We identified 910 RCTs that reported on fertility treatments, of which 182 RCTs (20%) reported on singleton live birth [before 2004 96/518 (19%); after 2003 86/392 RCTs (22%)]. Singleton live birth was the primary outcome in 68 RCTs (7.4%). Only 44 RCTs (4.8%) reported on neonatal outcome, while 52 RCTs (5.7%) reported on maternal outcome. We only included Cochrane reviews, thus report here only on the higher quality studies. The actual reporting on maternal and neonatal outcome may even be lower when studies of lower quality are included. Although a decade ago singleton live birth was recommended as the outcome measure of reproductive medicine research, this has not been followed; currently most clinical research in reproductive medicine does not report beyond the occurrence of pregnancy. No funding was received for the study. The authors have no conflicts of interest to declare.

Requirement for Mass Spectrometry Sex Steroid Assays in the Journal of Clinical Endocrinology and Metabolism

The accurate measurement of hormones is the pivot of modern endocrinology. Although tracing its roots to ancient writings, the modern scientific discipline came into existence after a series of mid-20th century Nobel Prizewinning achievements in identifying and measuring hormones. Arguably the most decisive step was the invention of immunoassay in 1959 (1), originally for peptide hormones but extended a decade later to nonimmunogenic steroids (2). This armed the nascent discipline with revolutionary impact through the ability to measure virtually every hormone at orders of magnitude lower than had ever been possible before with earlier cumbersome, insensitive, whole-animal bioassays. Yet the pioneers of steroid immunoassay understood its limitations as that decade’s delay was consumed in finding the additional workarounds required to create valid formats for immunoassay of small, subimmunogenic molecules like steroids. These essentials comprised solvent extraction, chromatography, and structurally authentic tracers, a triplet of validity criteria. Employing these required skills originally confined steroid measurement to manual methods in specialized laboratories; however, in the 1980–1990s, the steeply increasing demand for steroid measurement in the clinic and laboratory drove assay simplification to adapt steroid immunoassays into popular one-step kits and multiplex assay platform formats. This commodification resulted in “direct” steroid immunoassays, which bypassed all the original triplet of validity criteria (most fundamentally extraction), sacrificing accuracy and specificity for throughput speed and lower cost. Although problems with validity (analytical specificity, accuracy) of direct immunoassays were soon identified and were reported over 25 years ago (3, 4), the fundamental significance of violating these validity criteria resurfaced prominently in the last decade as the accuracy of direct hormone assays came under more stringent scrutiny—and as a solution, affordable and accessible mass spectrometry (MS) steroid assays came into view. The method-specific bias and analytical nonspecificity of direct T immunoassays are worst at the low circulating T levels in children, women, and men with pathological, functional, or experimentally induced T deficiency (5–7) where such direct immunoassays have been famously likened to random number generation (8). By modern standards, putative findings using such suboptimal methods would require verification by more specific MS-based assays. This renders direct T immunoassays methodologically inadequate for high-quality clinical reproductive medicine research. Not surprisingly, aiming to measure circulating (picomolar) estradiol concentrations 10to 100-fold lower than circulating (nanomolar) T levels that render direct T immunoassays unreliable is clearly problematic. Thus, direct estradiol immunoassays also lack accuracy in children (9), men (10, 11), and postmenopausal (3, 12–15) or aromatase inhibitor-treated (16) women who all share comparably low circulating estradiol concentrations. It is beyond doubt now that direct immunoassays of sex steroids are methodologically inadequate for high-quality clinical research. MS-based steroid measurements have been available for decades, indeed as long as immunoassays (17). Although they always remained the reference method for steroid specificity and structure, MS-based steroid assays were always too expensive and unavailable for routine use in the laboratory to support clinical research because they required highly skilled operators using complex, difficult-

Glycoprotein 130 promotes human blastocyst development in vitro

To investigate the efficacy of leukemia inhibitory factor (LIF) and/or glycoprotein 130 (gp130) on in vitro growth of human embryos. Laboratory study. University hospital-based IVF clinic. A total of 164 frozen embryos that survived thawing were cultured in media supplemented with LIF and/or gp130 or control media. Morphological development was evaluated by light microscopy. Protein expression profiles of single blastocysts were evaluated using matrix-assisted laser desorption/ionization time of flight-based intact cell mass spectrometry. Embryo development and protein content. Addition of gp130 to culture media improved blastocyst formation (73% vs. 43%). Addition of LIF to the culture media did not improve embryo development. Protein fingerprint spectra were obtained that revealed significant intensity changes for multiple molecular species including thymosin beta-10, thymosin beta-4, histone H2A, histone H2B, histone H4, ubiquitin, ubiquitin-T, and acyl-CoA binding protein. Glycoprotein 130, but not LIF, seems to be beneficial for preimplantation embryo development, implicating a physiological role in regulating preimplantation development in humans and thus ought to be included in culture media designed for embryo culture to the blastocyst stage. Furthermore, these findings highlight the great potential of matrix-assisted laser desorption/ionization time of flight mass spectrometry and intact cell mass spectrometry as a versatile tool in reproductive medicine research.

Qualitative research in reproductive medicine: From description to action

Abstract Assisted reproduction (ART), particularly that performed using donated gametes, increases the prospect of healthy babies being delivered to increasing numbers of people striving for parenthood. The psychosocial, ethical and legislative issues related both to the donation and receipt of gametes are perceived as extraordinarily complicated. In 2009, a research project aimed at mapping the issues was drawn up and implemented in the Czech Republic. The project should have provided material for consultation purposes, for the work of ethical and legislative bodies, and for better interdisciplinary and international communication in reproductive medicine. Work on the project was affected by several unforeseen events, particularly by the drafting and adoption of a new law on ART (to which the project was initially to have contributed material once concluded). The article describes the dynamic and structural changes occurring within the project due to drafting of the bill as well as the changes and consequences resulting from other circumstances related to the topic researched.

Effect of follicle stimulating hormone on the secretion of anti-mullerian hormone in the granulosa cells in patients with polycystic ovarian syndrome

To study the effect of follicle stimulating hormone (FSH) on the secretion of anti-mullerian hormone (AMH) in granulosa cells in patients with polycystic ovarian syndrome (PCOS). From Aug. 2008 to Dec. 2009, 33 patients with PCOS in Reproductive Medicine Research Center, Sixth Affiliated Hospital of Sun Yat-sen University were collected from 8-10 mm antral follicles and classified into the following three groups: FSH-unstimulated granulosa cells (unstimulated group, n = 12), exogenous FSH-stimulated granulosa cells (exo-stimulated group, n = 12) and internal FSH-stimulated granulosa cells (int-stimulated group, n = 21). ELISA and real-time PCR were relatively used to measure protein and mRNA level of AMH. Luciferase reporting vector was constructed to detect the promoter activity of AMH. The levels of AMH secretion in PCOS granulosa cells were (11.4 ± 4.0) µg/L in unstimulated group, (7.9 ± 1.1) µg/L in exo-stimulated group and (5.6 ± 1.7) µg/L in int-stimulated group. Both the external and internal stimulation of FSH may suppress AMH secretion significantly (P < 0.05). The mRNA level of AMH in PCOS granulosa cells were 2.5 ± 1.2 in unstimulated group, which were higher than 1.5 ± 0.5 in exo-stimulated group and 1.1 ± 0.7 in int-stimulated group significantly (P < 0.05). The luciferase activity of AMH in PCOS granulosa cells were 11.5 ± 2.3 in unstimulated group, 8.7 ± 2.4 in exo-stimulated group and 6.8 ± 2.4 in int-stimulated group. The luciferase activity of AMH in unstimulated group was significantly higher than those in exo-stimulated and int-stimulated groups (P < 0.05). FSH may inhibit the excessive secretion of AMH and stimulate follicle growth in PCOS granulosa cells by suppressing activity and expression of promoter.

Barriers to conducting clinical research in reproductive medicine: Australia and the United Kingdom

There are several major barriers to research in reproductive medicine in the United Kingdom and Australia. In Australia these mainly reflect income disparity between research and private practice whereas in United Kingdom they reflect the consequences of the economic downturn.

Barriers to conducting clinical research in reproductive medicine around the world

We asked seven senior and accomplished investigators from around the world to share their experiences conducting research in reproductive medicine.

Barriers to conducting clinical research in reproductive medicine: Latin America

Societies in Latin America are not scientifically driven and therefore, the allocation of human and economic resources to research is meager, as a reflection of this as well as other cultural and economic realities.

Barriers to conducting clinical research in reproductive medicine: India

Research in India is evolving at a rapid pace. Ergo, we need faster approval processes and review timelines. Improvement in the quality of institutional review boards would also contribute to good research practices. Clinical research should be included as a part of the undergraduate medical and pharmacy curriculum, and leading hospitals and practitioners should openly endorse research.

Barriers to conducting clinical research in reproductive medicine: China

In China, our research personnel and projects are mainly funded by the central government, provincial government, and nongovernmental foundations. Occasionally, we do some multicenter clinical research under the solicitation of pharmaceutical companies.

Barriers to conducting clinical research in reproductive medicine: Egypt

There are many barriers to scientific research in Egypt, including lack of funding, lack of a scientific atmosphere, and "brain drain" of scientists who produce high quality research.

Barriers to conducting clinical research in reproductive medicine: United States of America

A decrease in National Institute of Health funding and an increase regulations are significant barriers in conducting research in the United States of America.

Barriers to conducting clinical research in reproductive medicine: Greece

In Greece, procedures for obtaining funding and/or state approval for a research project are lengthy, hinder collaboration between centers, and may render projects outdated.

Minireview: The Roles of Small RNA Pathways in Reproductive Medicine

The discovery of small noncoding RNA, including P-element-induced wimpy testis-interacting RNA, small interfering RNA, and microRNA, has energized research in reproductive medicine. In the two decades since the identification of small RNA, first in Caenorhabditis elegans and then in other animals, scientists in many disciplines have made significant progress in elucidating their biology. A powerful battery of tools, including knockout mice and small RNA mimics and antagonists, has facilitated investigation into the functional roles and therapeutic potential of these small RNA pathways. Current data indicate that small RNA play significant roles in normal development and physiology and pathological conditions of the reproductive tracts of females and males. Biologically plausible mRNA targets for these microRNA are aggressively being discovered. The next phase of research will focus on elucidating the clinical utility of small RNA-selective agonists and antagonists.

Orthotopic testicular transplantation in mice

A method of revascularized orthotopic testicular transplantation (OTT) was developed in mice. The left testis was selected as donor graft for the operation due to less variation in anatomy. There were three groups: 1) a control group (n=24), 2) a group of castrated mice (n=24), and 3) a group in which OTT (n=24) was performed. Morphologically, the transplanted testes showed active spermatogenesis and normal structure of epididymis at 4 and 5 weeks. The function of the transplants was examined by RIA at designed time points. LH, FSH, and testosterone showed return to normal levels at 4 weeks. To our knowledge, this is the first report of successful revascularized OTT in mice. The model may prove useful in research in reproductive medicine, especially using knockout and transgenic mice.

The future potential of cryopreservation for assisted reproduction

Cryopreservation of human gonadal tissue and oocytes has brought about new and exciting research in reproductive medicine, as well as new cryopreservation techniques that are dramatically improving post-thaw viability and freezing damage. The work done on gonadal tissues is aimed at improving the quality of life for infertile patients and for prepubertal patients undergoing gonadotoxic chemotherapy, patients for whom hormonal stimulation/IVF is not an option, and women without partners. Cryopreservation of mature oocytes is the best model for timing IVF. Vitrification is likely to benefit the field, and since 2005, implantation and pregnancy rates from vitrified oocytes have matched or eclipsed results from conventional methods, due to new cell-specific methods and formulas. Cryopreservation of immature oocytes leads to a new direction of egg banking in future. Preserving ovarian tissue for autografting is still promising and has resulted in folliculogenesis, resumed hormone production and live births in limited cases. The use of small cortical blocks, or mechanical/chemical digestion of ovarian tissue for isolation of follicles is a new direction for ovary preservation for reasons of cryoprotectant permeation and graft revascularization. Maturation of follicles in vitro has become more feasible with the use of alginate microencapsulation. Testicular tissue preservation has taken a sharp turn towards preservation of gonadal stem cells. Research into the mechanism for spermatogenesis points to the ability for male germ cells to resume spermatogenesis. The cryopreservation of minced testicular tissue for isolation of germ cells via chemical digestion has produced viable cells, however, structural damage that may be avoided by vitrification has been noted to the surrounding cell junctions and supporting cells.

Keeping clinicians in clinical research: the Clinical Research/Reproductive Scientist Training Program

In recent years the need for translational and clinical research has increased, whereas the number of physicians involved in clinical research has diminished. There is clearly a need for formalized academic training in the quantitative and methodologic principles of clinical research in reproductive medicine. The Clinical Research/Reproductive Scientist Training Program (CREST), a program supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), the Clinical Research Training Program (CRTP) at Duke University, and the American Society for Reproductive Medicine (ASRM), meets this existing need. In addition, this program is specifically designed for physicians in private or academic clinical practice in reproductive medicine. Innovative programs such as CREST encourage the practicing physician to engage in clinical research while maintaining an active role in clinical practice. Participants in the program receive didactic on-line training from the CRTP and attend intensive weekend seminars at the National Institutes of Health (NIH) as well as CREST seminars at the annual meeting of the ASRM. Successful participants in the program receive a Certificate in Clinical Research from the CRTP. The program's goal is to provide practicing physicians with the tools and research credentials that will facilitate collaborations with investigators involved in large clinical trials.

Mass spectrometry‐based proteomics in reproductive medicine

The emergence of powerful mass spectrometry-based proteomic techniques has added a new dimension to the field of biomedical research. Application of these high throughput methodologies in pregnancy-related pathology has contributed to the comprehension of the underlying pathophysiologies and the successful identification of relevant protein biomarkers that can potentially change early diagnosis and treatment of several medical conditions related to human pregnancy. Most of the existing research on human reproduction and gestation has focused on follicular fluid, cervical/vaginal fluid, and amniotic fluid. Although proteome technologies in reproductive medicine research are not as yet widely applied, characterization of the proteome of reproductive fluids can be expected to significantly improve maternal healthcare. This article aims to summarize the applications of mass spectrometry based technology on the most important and specific biological fluids related to reproduction and gestation.