The hypo-osmotic swelling test: Is it a sperm vitality or a viability assay?

The first two editions of the World Health Organization laboratory manual described the determination of live spermatozoa by a dye exclusion method as a sperm "viability" test, whereas subsequent editions classified it as a "vitality" test, without providing an explanation for the reclassification. Additionally, the hypo-osmotic swelling (HOS) test, which assesses the functional integrity of the human sperm membrane, was placed in the same category as the dye exclusion test. Although the two terms might seem synonymous, the term "vitality" merely means "alive," whereas "viability" assesses qualities or physiological functions of a living entity.

Individual variation of the percentage of Y-chromosome bearing sperm content in human ejaculates.

The study aimed to determine the variation of Y-chromosome-bearing sperm content among individual ejaculates. A real-time polymerase chain reaction (qPCR) with unique primers was developed and used to calculate the percentage of Y-chromosome-bearing sperm in individual ejaculates from 50 randomly selected men. There was a significant difference in the overall mean ± SD between the proportion of Y-chromosome-bearing sperm and X-chromosome-bearing sperm (45.

A novel laboratory procedure to validate American Urological Association guideline on vasectomy success and to diagnose obstructive azoospermia.

The objective was to develop a laboratory procedure to validate American Urological Association (AUA) Guideline on vasectomy success when nonmotile spermatozoa are found in the post-vasectomy ejaculate. The neutral α-glucosidase (NAG) an epididymal protein assay modified to determine the activity at 30 and 90 min of incubation from 24 pre- and 47 post-vasectomy ejaculates. The difference between the two points in the relative activity was calculated and if the difference was nonsignificant will confirm vasectomy success.

Selecting the most competent sperm for assisted reproductive technologies.

This paper discusses the variety of effective sperm selection techniques that have been developed for use in assisted reproductive technologies. Available methods for isolating the competent sperm in an ejaculate are outlined, as well as techniques for selecting single sperm for use in intracytoplasmic sperm injection procedures. Case-specific methods for selecting the most competent sperm are discussed, with reference to the potential causes of male factor infertility and guidance for the embryologist based on the issues present for each couple seeking treatment.

Expression of a2 vacuolar ATPase in spermatozoa is associated with semen quality and chemokine-cytokine profiles in infertile men.

Background: A number of laboratory tests have been developed to determine properties of spermatozoa quality but few have been adopted into routine clinical use in place of the WHO semen analysis. We investigated whether Atp6v0a2 (a2 isoform of vacuolar ATPase) is associated with abnormal semen quality and changes in chemokine-cytokine profiles in infertile men.

Patients And Methods: Semen samples were collected from 35 healthy donors and 35 infertile men at the Andrology laboratory from August 2011 to June 2012.

The hypo-osmotic swelling test for evaluation of sperm membrane integrity.

A functional membrane is requisite for the fertilizing ability of spermatozoa, as it plays an integral role in sperm capacitation, acrosome reaction, and binding of the spermatozoon to the egg surface. The hypo-osmotic swelling (HOS) test evaluates the functional integrity of the sperm's plasma membrane and also serves as a useful indicator of fertility potential of sperm. The HOS test predicts membrane integrity by determining the ability of the sperm membrane to maintain equilibrium between the sperm cell and its environment.

Human chorionic gonadotropin from day 2 spent embryo culture media and its relationship to embryo development.

Objective: To detect hCG in spent embryo culture media at day 2 after intracytoplasmic sperm injection and to assess the relationship of hCG to embryo development.

Design: Experimental study.

Setting: Fertility center and clinical diagnostic laboratory.

Chromatin intact human sperm recovery is higher following glass wool column filtration as compared with density gradient centrifugation.

The sperm DNA quality as determined by chromatin integrity has been reported to be associated with in vivo and in vitro fertility. However, previous studies have evaluated preparation procedures to select motile, morphologically normal and mature spermatozoa, but not the spermatozoa with intact sperm chromatin. To determine which technique yields a population of spermatozoa with improved DNA quality, split ejaculate was processed with density gradient centrifugation (DGC) procedure and glass wool column filtration (GWF) procedure.

Is apolipoprotien E codon 112 polymorphisms associated with recurrent pregnancy loss?

Problem: To compare the prevalence of 112T>C point mutations among women experiencing RPL with fertile control women.

Method Of Study: Buccal swabs were obtained from 232 individuals: 136 with a history of >or=2 abortions, 37 with at least 2 live births and 59 with a history of deep vein thrombosis (DVT). DNA was extracted and PCR amplification of Apo E codons was performed.

Prevalence of antiphospholipid antibodies among women experiencing unexplained infertility and recurrent implantation failure.

The prevalences of antiphospholipid antibodies (APAs) among 1,325 women with a history of unexplained infertility and 676 women experiencing recurrent implantation failure were compared with 789 women experiencing recurrent pregnancy loss and 205 fertile control women. Eight percent and 9% of women with a history of unexplained infertility and recurrent implantation failure had more than one positive APA compared with 1.5% of fertile negative control women and 11% of positive control women experiencing recurrent pregnancy loss.

Are polymorphisms in the ACE and PAI-1 genes associated with recurrent spontaneous miscarriages?

Problem: To determine whether the ACE D/D genotype or the combination of PAI-1 4G/4G and ACE D/D genotypes may serve as a risk factor for recurrent pregnancy loss.

Method Of Study: Buccal swabs were obtained from 120 women experiencing recurrent pregnancy loss and from 84 fertile control women. DNA was extracted from the buccal swab samples using the Qiagen DNA Mini Kit (Qiagen), followed by multiplex polymerase chain reaction (PCR).

Correlation between neutral alpha-glucosidase activity and sperm DNA fragmentation.

To evaluate the association between neutral alpha-glucosidase (NAG) activity and sperm DNA fragmentation (DFI), ejaculates from 24 men undergoing evaluation for sperm DNA damage as a part of infertility assessment were analysed. The mean +/- SD and range for the semen quality of the 24 ejaculates are as follows: volume (3.1 +/- 1.

The relevance of neutral alpha-glucosidase activity in andrology.

To assess the usefulness of routine determination of neutral alpha-glucosidase (NAG) in andrology, 216 ejaculates were analyzed for NAG activity and semen quality. A correlation between NAG activity and semen volume and sperm concentration was determined; however, no correlation was observed between NAG activity and sperm motility or sperm morphology. The number of azoospermic ejaculates that had NAG activity below acceptable levels was significantly higher than the number of non-azoospermic ejaculates with similarly low NAG levels.

The association of Apoprotein E polymorphisms with recurrent pregnancy loss.

Problem: We have previously reported the role of polymorphisms of thrombogenic genes involved in coagulation and fibrinolysis as risk factors for recurrent pregnancy loss. Thrombophilia has been viewed as a multigenic disorder rather than a monogenetic clinical phenotype and Apo E has been shown to play an important role in lipid metabolism in pregnancy. As individuals carrying the E4 allele of the ApoE gene have the highest risk for thrombosis, we evaluated the frequency of the Apo E4 genotype among women suffering from recurrent pregnancy loss.

Thrombophilic gene polymorphisms are risk factors for unexplained infertility.

Earlier studies have shown inherited thrombophilia to be a risk factor for recurrent implantation failure, raising the question of whether this risk is related to the underlying cause of unexplained infertility or to the mechanisms involved in the implantation process. When nine thrombophilic gene polymorphisms were compared among 92 women with the diagnosis of unexplained infertility and 60 fertile control women, women with a history of unexplained infertility displayed a higher prevalence of MTHFR C677T polymorphisms than control women.

P53 tumor suppressor factor, plasminogen activator inhibitor, and vascular endothelial growth factor gene polymorphisms and recurrent implantation failure.

Objective: To examine whether a panel of gene polymorphisms, including p53 codon 72 tumor suppressor factor, plasminogen activator inhibitor 1 (PAI-1), and vascular endothelial growth factor (VEGF) -1154, are risk factors for implantation failure after IVF-ET.

Design: Case-control study.

Setting: Private laboratories and reproduction medical centers.

Comparison of thrombophilic gene mutations among patients experiencing recurrent miscarriage and deep vein thrombosis.

Problem: Inherited thrombophilia has been shown to be a risk factor for cardiovascular disease including deep venous thrombosis as well as reproductive disorders including recurrent pregnancy loss. We have previously reported three out of the 10 thrombophilic mutations studied, plasminogen activator inhibitor-1 (PAI-1) 4G/5G, factor XIII V34L, and homozygous MTHFR C667T, correlated significantly with recurrent pregnancy loss compared with controls. This study was undertaken to compare the frequencies of nine inherited thrombophilias among women with a history of recurrent pregnancy loss with individuals experiencing deep venous thrombosis and fertile controls.

Interleukin 1 receptor antagonist gene polymorphisms are not risk factors for recurrent pregnancy loss: evaluation of couples.

Problem: The interleukin-1 system has been implicated in pregnancy outcome. Fetal carriage of interleukin-1 receptor antagonist (IL-1Ra) specific alleles has been associated with adverse pregnancy outcomes including spontaneous abortion and pre-term labor. This study was undertaken to compare the frequency of IL-1RN 2 alleles among both male and female partners of couples experiencing recurrent pregnancy loss with that of fertile control couples.

Vascular endothelial growth factor gene polymorphism and implantation failure.

Implantation failure is the most frequent cause of lack of pregnancy after IVF and embryo transfer. Successful implantation requires the invading blastocyst to stimulate its own blood supply through angiogenesis. Vascular endothelial growth factor (VEGF) is the best-characterized regulator of angiogenesis.

Association of progesterone receptor polymorphisms with recurrent implantation failure after in vitro fertilization and embryo transfer.

Introduction: Progesterone is the hormone of pregnancy and is required for its initiation. The actions of progesterone are mediated by the progesterone receptor. Polymorphic variants of human progesterone receptor genes have been implicated in implantation failure.

Vascular endothelial growth factor gene polymorphisms and recurrent pregnancy loss.

Problem: To be successful, pregnancy must induce its own blood supply through angiogenesis and vascular endothelial growth factor (VEGF) is the best characterized regulator of angiogenesis and one polymorphism of the VEGF gene, -1154, has been suggested to be associated with recurrent spontaneous abortion. The aim of this study was to confirm or refute the relationship of VEGF -1154 to recurrent pregnancy loss (RPL).

Method Of Study: Buccal swabs were obtained from 152 women with history of two or more consecutive spontaneous abortions and 65 control women.

Cryopreservation of human sperm in a lecithin-supplemented freezing medium.

The use of egg yolk and serum albumin as additive diluents for human sperm cryopreservation is routine. But because both diluents are of animal origin, they potentially may introduce microbial agents to the sample. To reduce the risk of contamination, the cryoprotective property of phospholipids extracted from lecithin was evaluated and found to be effective when supplemented with dimethyl sulfoxide and glycerol.

The impact of male factor on recurrent pregnancy loss.

Purpose Of Review: The present paper reviews the current literature on the impact of male factor on recurrent pregnancy loss.

Recent Findings: Most clinicians focus their evaluation of recurrent pregnancy loss on the female, without much, if any, consideration of the other half of the couple - the male. Yet, the male contributes one-half of the genes for the embryo.

Discordance among blastomeres renders preimplantation genetic diagnosis for aneuploidy ineffective.

Purpose: To investigate the contribution of discordance among blastomeres from the same embryo in the interpretation of blastomeres biopsied from day 3 embryos.

Methods: 228 IVF embryos had two blastomeres removed and fluorescent in situ hybridization (FISH) was used to detect aneuploidy of chromosomes 13, 15, 16, 18, 21, 22, X and Y. Of the 228 embryos, 102 had complete FISH results for both blastomeres.