Density gradient centrifugation before or after magnetic-activated cell sorting: which technique is more useful for clinical sperm selection?

Purpose  

Although, at present, the selection of sperm prior to ICSI is based on motility and morphology, undetectable anomalies, and more importantly damaged DNA are overlooked. In this regard, novel sperm selection procedures have gained much interest. For instance, sperm has been selected by Magnetic-Activated Cell Sorting (MACS) based on early apoptotic marker, the externalization of phosphatidylserine (EPS). Review of the literature has revealed that the efficiency of this technique has been mainly evaluated post Density Gradient Centrifugation (DGC). Therefore, there is a need to prove the efficiency of this technique independent of DGC. In addition, considering the fact that DGC induces EPS due to capacitation and acrosome reaction, therefore, the role of MACS before DGC(MACS-DGC) and MACS after DGC (DGC-MACS) should be assessed.     

Annexin V magnetic-activated cell sorting versus swim-up for the selection of human sperm in ART: is the new approach better then the traditional one?

Purpose

To investigate whether the sperm fertilizing potential can be improved by selecting a non-apoptotic fraction using magnetic activated cell sorting (MACS), and to compare the results with the conventional swim-up method.

Methods

Twenty five male patients attending the andrology laboratory for sperm DNA fragmentation analysis. The sperm were prepared by density gradient centrifugation (DGC) and subsequently divided into three aliquots. The first was further separated into Annexin V-negative (non-apoptotic) fraction using MACS, the second was further processed by swim-up, while the third was left unseparated as a control. The impact of the combination of DGC with the two sperm preparation techniques on sperm quality was evaluated by comparing ‘rapid progressive’ motility, normal morphology according to Tygerberg’s strict criteria and DNA integrity (by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling [TUNEL]) for each aliquot.

Results

Sperm preparation that combines DGC with conventional swim-up method can provide sperm of higher quality in terms of motility, morphology and extent of DNA fragmentation compared to the Annexin V-negative (non-apoptotic) fraction derived from the combination of DGC with MACS.

Conclusions

Integrating MACS as a part of sperm preparation technique will not improve sperm fertilizing potential to the same extent as the traditional swim-up separation procedure.     

X-Y sperm selection: fact or fiction?

Selecting the gender of offspring has given rise to various and sometimes amusing stories. But regardless of which prefertilisation technique is used to influence the sex ratio of offspring it must fulfill certain criteria. First of all it must achieve a complete separation of the X and Y bearing sperm in sufficient quantities. Secondly sperm must be viable after separation and capable of fertilising. Sex preselection methods can be divided into two general groups which either separate spermatozoa on the basis of subtle physical or kinetic features or those which rely on distinctive nuclear characteristics unique either to X or Y chromosome bearing sperm. These, in turn, can be divided into in vivo methods designed to produce optimal conditions for fertilisation by either the X or Y bearing sperm, or in vitro sperm separation methods designed to separate X or Y bearing sperm. According to all published data, the different separation techniques have been shown not to be very effective. Only sex selection of spermatozoa by chromatin differences (cell sorting by flow cytometry) has demonstrated a significant enrichment of the X bearing sperm.     

Does endothelial cell activation occur with intrauterine growth restriction?

It is possible that in fetal growth restriction without pre-eclampsia endothelial cell activation does not occur. This might be either because there is no release of 'factor X' or because of maternal resistance to its effects. To test this hypothesis, we took blood samples from 26 women with pre-eclampsia (without fetal growth restriction), 13 women with fetal growth restriction (without pre-eclampsia) and 24 normal pregnant controls, and measured the circulating levels of three markers of endothelial cell activation (soluble VCAM, ICAM and E-selectin) and three cytokines [tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and -8 (IL-8)]. The levels of the markers of endothelial cell activation were raised in both pre-eclampsia and fetal growth restriction pregnancies compared with controls; however, the levels of TNF-α, IL-6 and IL-8 were significantly raised in pregnancies complicated by pre-eclampsia, but not in fetal growth restriction, compared with controls. These data show that endothelial cell activation is common to both pre-eclampsia and fetal growth restriction, but that the circulating levels of cytokines are elevated only in pre-eclampsia. Thus, it seems likely that endothelial cell activation is a consequence of a failure of trophoblast invasion and that a further step is required, possibly involving cytokine release, for the expression of the full clinical picture of pre-eclampsia.     

Does the timing of mock embryo transfer trial improve implantation in intracytoplasmic sperm injection cycles?

Aim.?To determine if endometrial trauma during embryo transfer trials (ETTs) improves implantation and clinical pregnancy rates in intracytoplasmic sperm injection treatment cycles.

Patients.?One-hundred fifty women undergoing their first IVF treatment were included in a prospective randomised study in a University hospital clinic. ETTs were performed either on day 21 of the previous cycle, or on day 6 of the controlled ovarian hyperstimulatin (COH) cycle, or conducted at least two cycles before COH cycle. Clinical pregnancy and implantation rates were compared between the groups.

Results.?There was no difference between the groups in terms of clinical pregnancy or implantation rates.

Conclusion.?Endometrial trauma by ETTs performed either during the preceeding cycle or on day 6 of the COH cycle does not improve pregnancy rates.     

Does training assist medical laboratory scientists with better evaluation of sperm morphology?

This study aimed to evaluate the quality of sperm morphology evaluation skills of 16 technologists who are responsible for semen analyses in their institution. The 1-day workshop was held at the Centre for Research in Reproductive Health in Sagamu, Nigeria. Participants were requested to complete a questionnaire that provided information regarding their experience and training as technologists in their institution. The workshop consisted of 4 sections, namely (i) pre-training test, (ii) lectures on morphometric characteristics and details of normal and abnormal sperm (iii) a laboratory-based hands-on, post training test and (iv) a consensus training session. The findings of the questionnaire indicated that none of the participants had any previous formal training and that all participants had very little knowledge about the morphological appearance of normal spermatozoa. These findings were supported by the results obtained during the pre-training session that showed a mean percentage difference of 24% between the reference and participant's values. These somewhat alarming results highlight the need for training facilities for technologists who work in the clinical diagnostic field of reproductive science.     

Does hyaluronan improve embryo implantation?

PURPOSE OF REVIEW: Taking into consideration the increasing interest on hyaluronan and its biological as well as physiological properties, this review will focus on the role of this molecule in human embryo implantation. RECENT FINDINGS: Several studies have been performed up to date in order to assess whether the addition of hyaluronan in the human embryo culture system can improve pregnancy and implantation rates, including one retrospective and six randomized controlled trials. On the one hand, four of those studies showed significant increase in clinical pregnancy and/or implantation rates after using embryo transfer medium containing high concentration of hyaluronan. On the other hand, three studies did not demonstrate any significant improvement in clinical pregnancy and implantation rates. However, regardless of statistical significance, almost all studies demonstrate higher pregnancy and implantation rates after using embryo transfer medium containing high concentration of hyaluronan. SUMMARY: Up to date, the results regarding the role of hyaluronan in human embryo implantation are still conflicting and, thus, further prospective randomized clinical trials are necessary to draw solid conclusions.     

Does sperm origin affect embryo morphokinetic parameters?

Purpose

The purpose of our study was to use time-lapse in order to evaluate the impact of sperm origin (fresh ejaculate or surgically retrieved) on embryo morphokinetic parameters and clinical outcome in intracytoplasmic sperm injection (ICSI) cycles.

Methods

This retrospective monocentric study was conducted in 485 unselected couples undergoing 604 ICSI cycles with embryo culture in the Embryoscope®. Among them, 445 couples underwent ICSI cycle with fresh ejaculated sperm and 40 with surgically retrieved sperm (26 with testicular sperm and 14 with epididymal sperm). Embryo morphokinetic parameters and clinical cycle outcome were compared between fresh ejaculated sperm and surgically retrieved sperm. A subgroup analysis was also conducted between testicular and epididymal sperm ICSI cycles.

Results

Clinical outcome was comparable between groups according to sperm origin. Although most early morphokinetic parameters were comparable between ejaculated and surgical sperm groups, a few parameters were significantly different between both groups, but with a considerable overlap in their distribution. Late cellular events occurred significantly later in the surgical sperm group than in the ejaculated sperm group.

Conclusions

Morphokinetic analysis did not allow us to identify clinically relevant differences between fresh ejaculate and surgically retrieved sperm groups. Further studies are needed, especially concerning the relationship between sperm origin and late morphokinetic parameters, such as blastocyst development.     

Does pelvic mesh treated with phosphorylcholine improve outcomes? An early experience

Objectives

Implantable devices treated with phosphorylcholine (PC) have been successfully used in cardiac, ophthalmic, and other applications. This surface modification has resulted in a reduction in the host inflammatory responses. This pilot study tested the safety and efficacy of PC treated polypropylene mesh grafts implanted for the treatment of pelvic organ prolapse.

Study design

Surgeons from five U.S. sites collected data on subjects implanted with Perigee IntePro Lite + PC. Pre-procedure data collected included demographics and prolapse severity. At follow-up, subjects were assessed for anatomical outcomes (success ≤ stage I POPQ or Baden Walker), symptomatic improvement, and complications, particularly mesh exposure.

Results

A total of 40 subjects were enrolled with 80% (32/40) of them completing at least 5–7 months of follow-up. Mean patient age was 60 years (range 36–78 years) and the mean BMI was 28 (range 20–40). There were no cases of mesh exposure/extrusion or granuloma formation. The anatomical success rate was 100% at 5–7 months (32/32).

Conclusions

This is the first publication on pelvic mesh treated with PC. There were no adverse events attributed to this surface modification. However, as the numbers are small, the results are not statistically significant. PC surface modification of pelvic mesh shows promise in its application for the reduction of mesh related complications.     

Does adjuvant chemotherapy improve survival for women with early-stage uterine leiomyosarcoma?

Objectives

To examine whether adjuvant therapy after primary surgery for treatment of early-stage uterine leiomyosarcoma (LMS) improves recurrence and survival rates.

Methods

A multisite, retrospective study of women diagnosed with stage I–II high grade LMS from 1990–2010 was performed. All patients (pts) underwent primary surgery followed by observation (OBS), radiotherapy (RT), or chemotherapy (CT) postoperatively.

Results

One hundred eight patients were identified with long-term follow-up; 94 pts (87.0%) had stage I and 14 (13.0%) had stage II disease. The mean patient age was 55.4 years and mean BMI was 28.0. Thirty-four (31.5%) patients underwent OBS, 35 (32.4%) received RT, and 39 (36.1%) received chemotherapy. After a median follow-up of 41.8 months, a recurrence was diagnosed in 70.8%. Recurrence was evident in 25/34 (73.5%) OBS, 23/35 (65.7%) RT, and 28/39 (71.8%) of CT cohorts and was not different based on treatment (p = 0.413). However, extra-pelvic recurrences were significantly higher in the RT (95.2%) than in the OBS (60%) or CT (64.3%) cohorts (p = 0.012). Additionally, recurrences were more likely to be successfully treated or palliated in those who initially received CT (p = 0.031). On multivariate analysis, stage (p < 0.001) and chemotherapy (p = 0.045) were associated with overall survival.

Conclusions

Women with early-stage, high grade uterine LMS experience high recurrence rates and poor survival outcomes, irrespective of adjuvant therapy. These rates are higher than previously reported in the literature. Although women treated with CT had similar recurrence rates as those treated with OBS or RT, treatment with adjuvant chemotherapy may decrease the risk of extra-pelvic recurrence and improve survival.     

Preconditioning of sperm with sublethal nitrosative stress: a novel approach to improve frozen–thawed sperm function

Salpingectomy is the most widely used treatment for hydrosalpinx. The effect of salpingectomy on the stimulation response during subsequent IVF treatment, however, remains unclear. The aim of this systematic review was to evaluate the ovarian response and pregnancy outcome of IVF treatment carried out after salpingectomy compared with other pre-IVF treatment options for hydrosalpinx. We conducted a literature search using PubMed, Ovid MEDLINE, Google Scholar, ClinicalTrials.gov and the Cochrane Central Register of Controlled Trials. Five randomized studies and nine observational studies were included in the systematic review and evaluated using Cochrane Collaboration's tool for randomized, Newcastle–Ottawa scale for observational studies and GRADE guidelines for certainty of evidence assessment. The mean number of retrieved oocytes was similar between the groups in randomized (mean difference [MD] = –0.03, 95% CI –0.75 to 0.70) and observational studies (MD = –0.15, 95% CI –2.32 to 2.02). Live birth (RR 1.59, 95% CI 1.17 to 2.16), clinical pregnancy (RR 1.27, 95% CI 1.02 to 1.57) and implantation rates (RR 1.55, 95% CI 1.16 to 2.08) were higher in the salpingectomy group in randomized studies. The present systematic review and meta-analysis showed that salpingectomy does not impair the ovarian response during subsequent IVF treatment.     

An artificial sperm – next year or never?

The idea that a sperm or egg might be formed outside of a gonad is perhaps as bizarre today as human assisted conception and test-tube babies were 30–40 years ago and has received a similar amount of media copy. Here, we review briefly the derivation of mammalian germ cells and sperm cells from embryonic stem cells in?vitro, ask whether these methods are viable and could provide therapies in the future, and discuss what constitutes an ‘artificial’ sperm.     

Does early morphology provide additional selection power to blastocyst selection for transfer?

Routine early developmental parameters are widely used in IVF centres to evaluate embryo development and fresh single-blastocyst transfer currently seems superior to single-embryo transfer. Would early morphological parameters help to choose the single blastocyst to be transferred, thereby improving the chances of implantation and live birth rate? This prospective observational study analysed the individual outcomes of 2617 embryos from 511 IVF couples scheduled for a single-blastocyst transfer. Embryo and blastocyst scores were constructed. There was a clear relationship between the kinetics and morphology of blastocysts and further implantation and live birth rate. There was a limited predictive value of embryo score with regard to blastocyst development and growth kinetics. Implanted and non-implanted blastocysts showed similar embryo scores. Thus usual morphological parameters on days 1 and 2 seem to have no additional value in indicating the right blastocyst to transfer. Non-invasive approaches might be helpful to increase the chances of implantation in the future.Routine early developmental parameters are widely used in IVF centres to evaluate embryo development, and fresh single-blastocyst transfer currently seems superior to single-embryo transfer. Would early morphological parameters help choose the single blastocyst to be transferred and thereby improve the chances of implantation and live birth rate? In this prospective observational study, we analysed the individual outcomes of 2617 embryos from 511 IVF couples scheduled for a single-blastocyst transfer. Embryo and blastocyst scores were constructed. There was a clear relationship between the kinetics and morphology of blastocysts and further implantation and live birth rate. There was a limited predictive value of embryo score with regard to blastocyst development and growth kinetics. Implanted and non-implanted blastocysts showed similar embryo scores. Thus usual morphological parameters on days 1 and 2 seem to have no additional value in indicating the right blastocyst to implant. Non-invasive approaches might be helpful to increase the chances of implantation in the future.     

Does antenatal physical therapy for pregnant women with low back pain or pelvic pain improve functional outcomes? A systematic review

Objective. A systematic review was undertaken to update the understanding of the available evidence for antenatal physical therapy interventions for low back or pelvic pain in pregnant women to improve functional outcomes when compared with other treatments or no treatment. Data Sources. Seven electronic databases were systematically searched and supplemented by hand searching through reference lists. Methods of study selection. Two reviewers independently selected trials for inclusion and independently assessed the internal validity of the included trials using the Clinical Appraisal Skills Program tool. Results. Four trials with 566 participants were identified that met the inclusion criteria. The validity of the trials was moderate. Exercise, pelvic support garments and acupuncture were found to improve functional outcomes in pregnant women with low back or pelvic pain. No meta-analysis was performed because of the heterogeneity of functional outcome measures. Conclusions. While there is some evidence that physical therapy using exercise, acupuncture and pelvic supports may be useful, further research needs to consider other treatment modalities used by physical therapists and establish an appropriate, reliable and valid functional outcome measure to assess low back and pelvic pain in pregnancy.     

Does bariatric surgery improve ovarian stimulation characteristics,oocyte yield,or embryo quality?

Background

Obesity is a major global health concern associated with multiple co-morbidities. Bariatric surgery has been considered a good treatment option in cases of morbid obesity. This preliminary study aims to investigate the effect of bariatric surgery on ovarian stimulation characteristics and IVF treatment cycle outcome.

Methods

A retrospective study that was performed in a tertiary, university-affiliated medical center and included all patients who underwent IVF treatment both before and after bariatric surgery. Data on ovarian stimulation variables of IVF treatment cycle prior and following the bariatric surgery were reviewed and compared.

Results

From January 2005 to June 2014, seven women fulfilled the inclusion criteria. After the operation, BMI was significantly reduced (mean?±?SD) (43.1?±?3.3 vs. 29.6?±?7.33, p?=?0.018), as was the number of gonadotropin ampoules required during stimulation (69.3?±?10.5 vs. 44.5?±?17, p?=?0.043). No between-cycle differences were observed in peak estradiol level, the number of oocytes retrieved, and percentage of mature oocytes.

Conclusions

To the best of our knowledge, this preliminary case series is the first comparison of IVF cycle characteristics prior to and following bariatric surgery. The operation seems to reduce treatment costs without affecting oocyte or embryo quality. Further large studies are required to establish the surgery’s effect on IVF outcome among infertile women.