Assessment of cell-free DNA and apoptosis in an oocyte microenvironment: promising biomarkers to predict intracytoplasmic sperm injection outcomes.

Cell-free DNA (cf-DNA) is defined as DNA fragments that are released into the body fluids from apoptosis or necrosis cells, including follicular fluid (FF), which can affect the microenvironment of the oocyte associated with infertility. We aimed to investigate a relationship between apoptosis of cumulus cells (CCs) and cf-DNA levels in FF and clinical outcomes of women undergoing intracytoplasmic sperm injection (ICSI). Therefore, 82 FF samples were collected, and the corresponding CCs were isolated for ICSI procedures. FF cf-DNA concentration was quantified using ALU-quantitative polymerase chain reaction (PCR), and CCs DNA fragmentation index (DFI) was evaluated by the terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labelling (TUNEL) method. We found that cf-DNA and DFI levels were significantly higher in FF and CCs samples related to the age of women ≥37 years compared with the age of women < 37 years. Moreover, in older and younger women, FF cf-DNA and CCs DFI levels were significantly lower when the anti-Müllerian hormone (AMH) level was > 1.1 ng/ml compared with when AMH ≤ 1.1 ng/ml. In addition, patients with a low number of retrieved oocytes ≤ 6 had significantly higher levels of CCs DFI and FF cf-DNA than women with a higher number of retrieved oocytes > 6. Additionally, we observed that higher levels of cf-DNA and DFI were associated with poor oocyte maturity and poor embryo quality. Finally, cf-DNA and DFI levels were significantly lower in pregnant women than in non-pregnant ones. We conclude that DFI and cf-DNA levels in the oocyte microenvironment could have potential use in evaluating oocyte and embryo developmental competence.

The Impact of Origanum vulgare Supplementation on Human Asthenozoospermic Sperm Parameter Quality.

Male infertility is a complex multifactorial disease and a real health problem; 50% of infertile men have identifiable causes detectable by fundamental sperm analysis. Numerous research studies have shown the possibility of treating abnormal semen samples with some drugs before artificial insemination, yet evidence of the drug's effectiveness remains minimal. In our previous work, we tested the effectiveness of some essential oils, such as eucalyptus (Eucalyptus globulus Labill.), oregano (Origanum vulgare L.), and sage (Salvia officinalis L.) on sperm parameters. The essential oil of oregano showed the best ameliorative effect. In present, we examined the effect of the essential oil of O. vulgare on the physiological parameters and the specific activity of certain antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), lipid peroxidation rate (MDA), and GAPDH and antioxidant and metabolic biomarkers, characterizing the quality of human sperm. The results showed that in vitro supplementation of oregano significantly improves the mobility and antioxidant activities, without harmful effects on the integrity of the sperm's DNA, and that the selected concentration of oregano EO is nontoxic and may be considered a therapeutic alternative to heal sperm motility problems in asthenozoospermic patients.

Women with extreme low AMH values could have in vitro fertilization success.

Circulating anti-müllerian hormone (AMH) and antral follicle count (AFC) are the best predictors of IVF outcomes. However, in extreme low AMH range especially for young patients, AMH prediction power loose its specificity to give real idea of pregnancy chance with IVF treatments and good prognosis of an extremely reduced ovarian reserve and expected poor response. Indeed, this retrospective study was conducted to evaluate IVF outcomes in patients following IVF-ICSI program with extremely low AMH levels (≤0.4 ng/ml; n = 390) compared to those presenting normal AMH range (1.3-2.6 ng/ml; n = 352) considered as control group. As expected, number of oocytes retrieved per patient, and embryological outcomes were significantly lower in the extremely low AMH levels group compared to control. Moreover, it was same trend concerning clinical outcomes but we have to note that even in extreme low AMH, patients could reach ineligible satisfying clinical pregnancy rate compared to control (17% vs 41%). For patients younger than 35 years, clinical pregnancy rate improved to 27%. Women with extreme low AMH values and especially younger ones, still have reasonable chances of achieving pregnancy, highlighting the default view of this category generally excluded from IVF program.

Luteal estradiol pretreatment of poor and normal responders during GnRH antagonist protocol.

Luteal estradiol pretreatment (LEP) to IVF protocols designed to improve follicle synchronization and retrieval of mature oocytes. We conducted a retrospective study including women undergoing IVF program who were given a course of 4 mg oral estradiol-17ß daily from day 20 of the same cycle until day 1 of their next cycle before starting an antagonist protocol, forming LEP-group but control-group started on day 3 a stimulation without pretreatment. A total is divided into 2 groups (poor (group 1, n = 148) and normal responders (group 2, n = 244)). Our findings show for group 1 a significant decrease in cancelation rate (3% vs 14%) and a significant improvement in clinical outcomes (clinical pregnancy per transfer and live birth rate respectively: 47% and 44% vs 12% and 11%). For group 2, this pretreatment could increase significantly the maturation rate (77% vs 68%). The rate of frozen embryos was improved in both groups: (group 1: 11% vs 2% and group 2: 53% vs 41%). LEP increases the frozen embryos rate whatever the nature of the ovarian response, but especially for normal responders it coordinates follicular recruitment increasing the maturation rate. In the case of poor responders, it affects positively clinical outcomes decreasing the canceled cycles.

The Effect of Supplementation with Some Essential Oils on the Mobility and the Vitality of Human Sperm.

The objective of this work is to study the improvement effect of some essential oils of sage (Salvia officinalis), oregano (Origanum vulgare), and eucalyptus (eucalyptus globulus) on the physiological parameters characterizing the quality of human sperm (mobility and vitality). We find natural biomolecules to improve sperm quality to increase the chances of success of very low in vitro fertilization (IVF) that stagnate around 20%. Sperm samples were mixed with different concentrations of essential oils. The effect of these essential oils on the motility and vitality of spermatozoa has been analyzed. The mobility was determined using a Computer Assisted Sperm Analysis (CASA). In the other side, the evaluation of sperm vitality was performed by staining eosin 2% and the microscopic examination is carried out via optical microscope. A drop of sperm will be mixed with a drop of eosin solution 2%, spread between the slip and coverslip, then allowed to air dry, and examined under a microscope. A significant improvement in the mobility and vitality of human spermatozoa has been noted with oregano. Eucalyptus after 10 min of exposure also significantly improves the mobility and vitality of the spermatozoa. Sage does not improve mobility for these incubation times but significantly improves vitality.

Paternal age: Negative impact on sperm genome decays and IVF outcomes after 40 years.

This study assessed sperm quality declining on relation to paternal age and its impact on in vitro fertilization (IVF) outcomes in order to estimate the APA (Advanced Paternal Age) cutoff. For this, 83 couples undergoing IVF treatment for male factor infertility were enrolled. The women age was ≤39 years, whereas the men were divided in two groups: APA (n = 41; age ≥ 40 years) and young (Y) (n = 42; age < 40 years). Conventional semen parameters (volume, concentration, motility, vitality, and morphology) were analyzed in the collected sperm samples. Furthermore, sperm genome decays (SGD) was assessed by TUNEL assay (DNA fragmentation), aniline blue staining (chromatin decondensation), and fluorescent in situ hybridization (aneuploidy). No significant difference was found concerning the conventional semen parameters between APA and Y groups. Conversely, SGD analysis showed increased DNA fragmentation; chromatin decondensation and sperm aneuploidy rates in the APA group (respectively, 41%, 43%, and 14% vs. 25%, 23%, and 4% in Y group). IVF outcomes also were affected by paternal age as indicated by the rates of cancelled embryo transfers, clinical pregnancy and miscarriage in the two groups APA and Y (29%, 17%, and 60% vs. 10%, 32%, and 42%). Finally, statistical analysis of the results suggests that the age of 40 should be considered as the APA cutoff during ART attempts.

Intrauterine insemination of cultured peripheral blood mononuclear cells prior to embryo transfer improves clinical outcome for patients with repeated implantation failures.

Implantation failure is a major limiting factor in assisted reproduction improvement. Dysfunction of embryo-maternal immuno-tolerance pathways may be responsible for repeated implantation failures. This fact is supported by immunotropic theory stipulating that maternal immune cells, essentially uterine CD56+ natural killer cells, are determinants of implantation success. In order to test this hypothesis, we applied endometrium immuno-modulation prior to fresh embryo transfer for patients with repeated implantation failures. Peripheral blood mononuclear cells were isolated from repeated implantation failure patients undergoing assisted reproductive technology cycles. On the day of ovulation induction, cells were isolated and then cultured for 3 days and transferred into the endometrium cavity prior to fresh embryo transfer. This immunotherapy was performed on 27 patients with repeated implantation failures and compared with another 27 patients who served as controls. Implantation and clinical pregnancy were increased significantly in the peripheral blood mononuclear cell test versus control (21.54, 44.44 vs. 8.62, 14.81%). This finding suggests a clear role for endometrium immuno-modulation and the inflammation process in implantation success. Our study showed the feasibility of intrauterine administration of autologous peripheral blood mononuclear cells as an effective therapy to improve clinical outcomes for patients with repeated implantation failures and who are undergoing in vitro fertilization cycles.

From global proteome profiling to single targeted molecules of follicular fluid and oocyte: contribution to embryo development and IVF outcome.

The development of in vitro fertilization (IVF) techniques for infertility management has led to the investigation of the proteome of follicular fluid and oocyte. In addition, different markers contributing to oocyte maturation and embryo development potential have been reported in the literature. Different techniques were utilized to analyze whole proteome or single protein markers in follicular fluid and oocytes, particularly in animal models. Data from several studies have generated large amounts of information, however, an ideal profile to predict the best oocytes and embryos suitable for implantation are still to be uncovered. The identification of such profiles and markers from follicular fluid, oocytes and endometrium should help scientists and clinicians develop better strategies to improving clinical outcome of IVF cycles.

Impact of sperm genome decay on Day-3 embryo chromosomal abnormalities from advanced-maternal-age patients.

Infertile male patients often exhibit unconventional semen parameters, including DNA fragmentation, chromatin dispersion, and aneuploidy-collectively referred to as sperm genome decay (SGD). We investigated the correlation of SGD to embryo chromosomal abnormalities and its effect on clinical pregnancy rates in patients with advanced maternal age (AMA) (>40 years) who were undergoing intracytoplasmic sperm injection-preimplantation genetic screening (ICSI-PGS). Three groups were assessed: patients with AMA and male partners with normal sperm (AMA-N); AMA patients and male partners presenting with SGD (AMA-SGD); and young fertile female patients and male partners with SGD (Y-SGD). We found a significant increase in embryonic chromosomal abnormalities-polyploidy, nullisomy, mosaicism, and chaotic anomaly rates-when semen parameters are altered (76% vs. 67% and 66% in AMA-SGD vs. AMA-N and Y-SGD groups, respectively). Statistical analysis showed a correlation between SGD and aneuploidies of embryonic chromosomes 13, 16, 21, X, and Y, as well as negative clinical outcomes. Incorporation of molecular sperm analyses should therefore significantly minimize the risk of transmission of chromosomal anomalies from spermatozoa to embryos, and may provide better predictors of pregnancy than conventional sperm analyses. We also demonstrated that an ICSI-PGS program should be implemented for SGD patients in order to limit transmission of chromosomal paternal anomalies and to improve clinical outcome.

Globozoospermia is mainly due to DPY19L2 deletion via non-allelic homologous recombination involving two recombination hotspots.

To date, mutations in two genes, SPATA16 and DPY19L2, have been identified as responsible for a severe teratozoospermia, namely globozoospermia. The two initial descriptions of the DPY19L2 deletion lead to a very different rate of occurrence of this mutation among globospermic patients. In order to better estimate the contribution of DPY19L2 in globozoospermia, we screened a larger cohort including 64 globozoospermic patients. Twenty of the new patients were homozygous for the DPY19L2 deletion, and 7 were compound heterozygous for both this deletion and a point mutation. We also identified four additional mutated patients. The final mutation load in our cohort is 66.7% (36 out of 54). Out of 36 mutated patients, 69.4% are homozygous deleted, 19.4% heterozygous composite and 11.1% showed a homozygous point mutation. The mechanism underlying the deletion is a non-allelic homologous recombination (NAHR) between the flanking low-copy repeats. Here, we characterized a total of nine breakpoints for the DPY19L2 NAHR-driven deletion that clustered in two recombination hotspots, both containing direct repeat elements (AluSq2 in hotspot 1, THE1B in hotspot 2). Globozoospermia can be considered as a new genomic disorder. This study confirms that DPY19L2 is the major gene responsible for globozoospermia and enlarges the spectrum of possible mutations in the gene. This is a major finding and should contribute to the development of an efficient molecular diagnosis strategy for globozoospermia.

Evaluation of Oxidative and Nitrosative Stress Markers Related To Inflammation in The Cumulus Cells and Follicular Fluid of Women Undergoing Intracytoplasmic Sperm Injection: A Prospective Study.

BACKGROUND: Oxidative/nitrosative stress in the oocyte microenvironment could have an impact on intracytoplasmic sperm injection (ICSI) outcomes. The presence of reactive oxygen species (ROS) and reactive nitrogen species (RNS) can stimulate the secretion of pro-inflammatory cytokines, leading to chronic inflammation and potentially affecting embryo as well as oocyte quality. This study aimed to examine the relationship of lipid peroxidation [measured by the malondialdehyde (MDA) assay] with protein carbonyl [measured by the 2,4 dinitrophenylhydrazine (DNPH) assay] levels in cumulus cells (CCs), as well as nitric oxide (NO), peroxynitrite (ONOO-), and C-reactive protein (CRP) levels in follicular fluid (FF). The potential relationship of these levels with ICSI outcome was also evaluated. MATERIALS AND METHODS: In this prospective study, 63 FF samples and their corresponding CCs were collected for ICSI procedures. Spectrophotometry was used to assess levels of DNPH, MDA, NO, and ONOO-. CRP levels were evaluated using an immunoturbidimetric assay. RESULTS: The patients under 37 years with normal ovarian reserve had significantly lower levels of MDA, DNPH, NO, ONOO-, and CRP compared to those over 37 years. Additionally, we observed higher levels of MDA, DNPH, NO, ONOO-, and CRP in the group with an oocyte maturity rate of less than 60%. No significant difference was observed between the DNPH levels and factors such as infertility duration, embryo quality, pregnancy, or the number of retrieved oocytes. A higher level of MDA, NO, ONOO-, and CRP was found to be significantly related to the lower number of retrieved oocytes, longer periods of infertility, poor embryo quality, and negative pregnancy outcomes. CONCLUSION: Oxidative/nitrosative stress, linking to inflammation in the oocyte microenvironment, can be considered as a potentially useful biomarker for assessing the development and competence of oocytes and embryos and predicting ICSI outcomes.

Genome and Epigenome Disorders and Male Infertility: Feedback from 15 Years of Clinical and Research Experience.

Infertility affects around 20% of couples of reproductive age; however, in some societies, as many as one-third of couples are unable to conceive. Different factors contribute to the decline of male fertility, such us environmental and professional exposure to endocrine disruptors, oxidative stress, and life habits with the risk of de novo epigenetics dysregulation. Since the fantastic development of new "omes and omics" technologies, the contribution of inherited or de novo genomes and epigenome disorders to male infertility have been further elucidated. Many other techniques have become available to andrology laboratories for the investigation of genome and epigenome integrity and the maturation and the competency of spermatozoa. All these new methods of assessment are highlighting the importance of genetics and epigenetics investigation for assisted reproduction pathology and for supporting professionals in counselling patients and proposing different management strategies for male infertility. This aims to improve clinical outcomes while minimizing the risk of genetics or health problems at birth.

The Addition of Opuntia ficus-indica Ethanolic Extract to a Skimmed Milk-Based Extender Impacts Ram Sperm Quality.

Recently, researchers have focused on the use of natural antioxidants to improve semen quality as a key element for successful artificial insemination. In this context, the first aim of this study was to determine the antioxidant activity and composition (minerals, vitamins, and sugars) of Opuntia ficus-indica cladode ethanolic extract (ETHEX). A further purpose of the study was to investigate the effect of ETHEX supplementation on the quality of liquid ram semen extended with skim milk (SM) at 5°C. The antioxidant activity of ETHEX was studied using free radical 1, 1-diphenyl-2-picrylhydrazyl (DPPH•) assay. The mineral composition and the sugar and vitamin contents of ETHEX were determined using an inductively coupled plasma optical emission spectrometry (ICP-OES) and HPLC-DAD-RID analytical instruments. As a second part, semen was collected from five Boujaâd rams with an artificial vagina. The ejaculates with more than 70% motility were pooled, extended with skim milk (SM) extender without (control) or supplemented with 1-8% of ETHEX (37°C; 0.8 × 109 sperm/mL). Sperm quality parameters were assessed at 8, 24, 48, and 72 h. The results showed that ETHEX had a higher antioxidant activity compared to those of ascorbic acid and butylated hydroxytoluene (BHT). Furthermore, ETHEX contains a considerable amount of minerals, vitamins, and sugars. The inclusion of 1 or 2% ETHEX in SM increased the sperm motility, viability, and membrane integrity and decreased the abnormality of spontaneous and catalyzed lipids peroxidation (p < 0.05) up to 72 h. In addition, semen diluted with 1 and 2% ETHEX decreased the level of DNA fragmentation compared to the control group (p < 0.05). In conclusion, the ETHEX could be recommended to improve the quality of liquid ram spermatozoa. However, its effects on artificial insemination should be further studied.

Effect of Origanum vulgare Essential Oil Supplementation on the Advanced Parameters of Mobility and on the Integrity of Human Sperm DNA.

The reduced sperm mobility is one of the most important causes of male infertility. Several reports have indicated that the treatment of subnormal sperm samples with certain agents prior to artificial insemination significantly improves the fertilizing potential of sperm. We have among others some stimulants such as pentoxifylline, relaxin, prostaglandin E, and diltiazem. In our precedent work, we had tested the effect of supplementation with three essential oils, namely, sage (Salvia officinalis), oregano (Origanum vulgare), and eucalyptus (Eucalyptus globulus), on sperm cell mobility and vitality. Oregano oil had shown interesting biological properties by giving the best values of progressive mobility and vitality. In this study, we aim to verify the effect of oregano oil supplementation on the advanced parameters of mobility and on the integrity of the sperm DNA of 25 male infertile volunteers. Our results showed that oregano oil over an incubation period of 5 to 10 min of exposure significantly improves the advanced parameters of mobility, namely, curvilinear velocity (VCL), linear velocity (VSL), the mean velocity of the path (VAP), and the amplitude of the displacement (ALH). The effect of the increase in the VCL decreased the linearity (LIN), the mean line (STR), and the mean wobble (WOB). Oregano oil at 5 min had no significant effect on the DNA fragmentation index (DFI) and sperm decondensation index (SDI). However, at 10 min, it had a significant effect on both DFI and SDI. The analysis of our results showed that this plant oil rich in terpenoids and phenolic antioxidants could be a quite good in vitro additive with high potential for the world of medically assisted reproduction.

Follicular fluid and supernatant from cultured cumulus-granulosa cells improve in vitro maturation in patients with polycystic ovarian syndrome.

OBJECTIVE: To study the effectiveness of a new in vitro maturation (IVM) approach based on heterologous follicular fluid (HFF) and supernatant of cumulus-granulosa cells (CGCs) mimicking the intact follicular microenvironment to rescue immature denuded oocytes (IDOs) of patients with polycystic ovary syndrome (PCOS) whose IVM or IVF outcomes remain poor. DESIGN: Randomized controlled trial. SETTING: University-affiliated private center. PATIENT(S): One hundred fifty-nine IDOs were obtained from 47 patients with PCOS. First, a simple IVM system (S-IVM; 40 IDOs; control group) was compared with different protocols based on the addition of autologous follicular fluid (AFF-IVM; 44 IDOs), HFF (HFF-IVM; 42 IDO), or HFF with CGC isolated from seven women without PCOS and presenting 100% in vivo oocyte maturation (HFF/CGC-IVM; 33 IDOs). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): IVM outcomes were compared among the four groups (S-IVM, AFF-IVM, HFF-IVM, HFF/CGC-IVM); then the vitro and in vivo maturation results (from controlled ovarian stimulation of PCOS patients) were compared for each group. RESULT(S): The HFF/CGC-IVM method gave the best yield of developed blastocysts per IDO compared with S-IVM, AFF-IVM, and HFF-IVM (27% vs. 2%, 2%, and 12%, respectively). The IVM rate with the HFF/CGC-IVM method was even higher than that compared with the in vivo maturation rate (79% vs. 42%), with significant improvement in the cleavage rate (71% vs. 61%). CONCLUSION(S): This adapted IVM system could be used to reach an acceptable result in meiotic competence and competent metaphase II oocytes capable of developing into intact embryos after fertilization and before transfer.

AZF microdeletions and partial deletions of AZFc region on the Y chromosome in Moroccan men.

AIM: To evaluate for the first time the frequency of Y chromosome microdeletions and the occurrence of the partial deletions of AZFc region in Moroccan men, and to discuss the clinical significance of AZF deletions. METHODS: We screened Y chromosome microdeletions and partial deletions of the AZFc region of a consecutive group of infertile men (n = 149) and controls (100 fertile men, 76 normospermic men). AZFa, AZFb, AZFc and partial deletions of the AZFc region were analyzed by polymerase chain reaction (PCR) according to established protocols. RESULTS: Among the 127 infertile men screened for microdeletion, four subjects were found to have microdeletions: two AZFc deletions and two AZFb+AZFc deletions. All the deletions were found only in azoospermic subjects (4/48, 8.33%). The overall AZFc deletion frequency was low (4/127, 3.15%). AZF microdeletions were not observed in either oligoasthenoteratozoospermia (OATS) or the control. Partial deletions of AZFc (gr/gr) were observed in a total of 7 of the 149 infertile men (4.70%) and 7 partial AZFc deletions (gr/gr) were found in the control group (7/176, 3.98%). In addition, two b2/b3 deletions were identified in two azoospermic subjects (2/149, 1.34%) but not in the control group. CONCLUSION: Our results suggest that the frequency of Y chromosome AZF microdeletions is elevated in individuals with severe spermatogenic failure and that gr/gr deletions are not associated with spermatogenic failure.

Emerging diagnostic, prognostic and therapeutic biomarkers for ovarian cancer.

BACKGROUND: In spite of various treatment options currently available, ovarian cancer (OC) still remains a leading cause of death in women world-wide. Diagnosis at an early stage is one of the most important factors that determines survival. Current clinical diagnostic tools have, however, a limited efficacy in early OC detection. Therefore, there is a critical need for new (early) diagnostic biomarkers and tools. Through advances in genomic, proteomic and metabolomic techniques, several novel molecular OC biomarkers have recently been identified. These biomarkers are currently subject to validation. In addition, integration of genomic, proteomic and metabolomic data, in conjunction with epidemiologic and clinical data, is considered essential for obtaining useful results. Interesting recent work has already shown that specific diagnostic biomarkers, such as BRCA mutations, may have profound therapeutic implications. Here, we review the current state of OC research through literature and database searches, with a focus on various recently identified biomarkers via different technologies for the (early) diagnosis, prognosis and treatment of OC. CONCLUSIONS: Multi-biomarker panels accompanied by a meticulous determination of their sensitivity and specificity, as well their validation, using multivariate analyses will be critical for its clinical application, including early OC detection and tailor-made OC treatment.

Effect of argan oil on liquid storage of ram semen in Tris or skim milk based extenders.

Due to its high antioxidant content, the argan oil could play a beneficial role in liquid storage of ram semen. The aim of this study was to investigate effects of different concentration of argan oil (ARO) on spermatologic parameters, lipid peroxidation and DNA fragmentation during liquid storage of ram semen until 48 h. Also effects of extenders and temperature on same parameters were assessed. For these aims, semen samples were collected from Boujaâd rams, extended with Tris egg yolk or skim milk extenders without (control) or supplemented with different concentrations of ARO (1%, 2%, 5% and 10% v/v) at a final concentration of 0.8 × 10(9) sperm/mL and stored until 48 h at 5 °C or 15 °C. The sperm quality assessments were performed at different intervals during storage (0, 8, 24 and 48 h). Sperm progressive motility started to decrease after 8h of storage in all temperatures--extenders combinations and dropped steadily during the 8-48 h interval. However, sperm viability, progressive motility and membrane integrity were markedly higher in ARO groups (especially in 1% in Tris and 5% in skim milk) until 24h and 48 h storage at both temperatures compared to controls. The argan oil also decreased the level of spontaneous and induced malondialdehyde (MDA) and the sperm DNA fragmentation until 48 h storage. In conclusion, it was determined that addition of argan oil to conventional extenders may improve the quality of ram semen during liquid storage in different temperatures.

Mitochondria: participation to infertility as source of energy and cause of senescence.

Mitochondria is a powerhouse organelle involved in ATP synthesis, calcium signaling, reactive oxygen species (ROS) by oxidative stress production, cell cycle arrest via apoptosis and sex steroid hormones biosynthesis. Improvement of sperm parameters such as motility, capacitation, acrosome reaction, and oocyte interaction, involve regulation of ROS levels by the mitochondria. In human, the relation between the quantitative level of mitochondrial DNA (mtDNA), oocyte cytoplasm maturation and fertilization potential, is not clear. It has been hypothesized that oocytes without sufficient wild type mtDNA and therefore able to generate ATP, would not normally be ovulated. This is reflected in the low numbers of mtDNA observed in degenerate oocytes obtained through super ovulation protocols during assisted reproductive technology programs. Different theories place mitochondria in a central role of oxidative damage to cells and tissues related to infertility declining and aging. Mitochondria-dependent apoptosis seems to be responsible for the pre and post-natal decline in germ cells, embryo development, implantation failure, and miscarriages.

Prevalence of the Aurora kinase C c.144delC mutation in infertile Moroccan men.

OBJECTIVE: To evaluate the carrier frequency of the pathogenic c.144delC mutation in AURKC gene and the contribution of this mutation in male infertility in a Moroccan population. DESIGN: Sanger sequencing of exon 3 in AURKC gene in infertile and control patients in Morocco. SETTING: Research institute. PATIENT(S): A total of 326 idiopathic infertile patients, and 450 age-related men. INTERVENTION(S): The incidence of AURKC c.144delC mutation was determined in men with unexplained spermatogenic failure and a control cohort of normospermic fertile men. MAIN OUTCOME MEASURE(S): Genomic DNA was extracted from peripheral blood lymphocytes and the screening of the c.144delC mutation in AURKC gene performed by polymerase chain reaction and sequencing. RESULT(S): The c.144delC mutation in AURKC gene was found in patients at homozygous and heterozygous states, with an allelic frequency of 2.14%, whereas in controls this mutation was found only in the heterozygous state, with lower frequency (1%). Homozygous patients were characterized by macrocephalic and multiflagellar spermatozoa. CONCLUSION(S): Our data indicate that the AURKC c.144delC mutation has a relatively high carrier frequency in the Moroccan population; thus, we recommend screening for this deletion in infertile men with a high percentage of large-headed and multiflagellar spermatozoa.