Leucocyte subpopulations in the seminal plasma and their effects on fertilisation rates in an IVF cycle.

Controversy exists on the role of leucocytospermia on fertilisation rates and IVF outcomes. The aim of our study was to identify the effect of leucocytes and leucocyte subpopulations on fertilisation rates in an IVF cycle. A prospective comparative study of the leucocyte subpopulations of seminal fluid of partners of women attending an IVF cycle was conducted. The samples underwent immunocytochemical staining. The monoclonal antibodies used in this study include CD3, CD4, CD8 (T Cells), CD14 (monocytes/macrophages), CD16 (granulocytes), CD20 (B Cells), CD45 (Pan Leucocytes), CD56 (natural killer cells) and CD69 (activated T and B Cells). Of 21 patients who were recruited into the study, seven were identified as poor fertilisers (<35%) and 14 were identified as good fertilisers (>60%). Data were analysed with SPSS version 14. The total leucocyte counts (CD45) between the poor and good fertilisers were not statistically significant. The macrophages and the monocytes (CD14) were significantly elevated in the good fertilisers group in comparison with the poor fertilisers (P < 0.05). We also found that T cells (CD2, CD4, CD8) and CD14 (macrophages) correlated significantly (r = 0.47, P value < 0.01) with the fertilisation rate. Our study confirms that the presence of leucocytes does not adversely affect the fertilisation rates and the outcome of an IVF cycle. However, macrophages and the monocytes (CD14) were significantly elevated in the good fertilisers group. The increased phagocytic activity in these individuals might increase their fertilising potential by removing spermatozoa with abnormal morphology.

Does repeated electro-ejaculation improve sperm quality in spinal cord injured men?

STUDY DESIGN: Retrospective analysis. OBJECTIVES: To assess the effect of repeated electro-ejaculation on the sperm quality in spinal cord injured men. SETTING: Regional Spinal Injuries Unit, Southport, UK and Hewitt Center for Reproductive Medicine, Liverpool, UK. METHOD: Retrospective, observational study of men with spinal cord injuries undergoing repeated electro-ejaculation as a part of fertility treatment. RESULT: There was no improvement in the volume, sperm concentration, motility or the total motile count in the successive antegrade and retrograde samples following repeated electro-ejaculations. CONCLUSION: Electro-ejaculation is an invasive procedure and its use should be restricted to obtaining semen sample for carrying out assisted conception procedures only.

Growing concern over the safety of using mobile phones and male fertility.

There are growing concerns about the possible hazards of electromagnetic waves emitted by mobile phones on human health. One of the biggest concerns is their possible association with increased risk of cancer and their possible effects on cellular DNA. Electromagnetic waves can inflict their results through both thermal and non-thermal effects. There are many animal studies that show that electromagnetic waves have a wide range of damaging effects on the male reproductive system and sperm parameters. However, similar studies are quite limited in humans, and the results of animal studies should be interpreted with caution when considering their application to humans. Large controlled studies are required before confirming such possible effects on male fertility.

A case of rapid deterioration in sperm quality following spinal cord injury.

STUDY DESIGN: Case report. SETTING: Northwest Regional Spinal Injuries Unit, Southport, UK and Reproductive Medicine Unit, Liverpool Women's Hospital, Liverpool, UK. CASE REPORT: A 28-year-old man suffered from paraplegia on sustaining a fall. Fertility preservation was an important aspect of his treatment and electro-ejaculation and cryopreservation of sperm was possible within days of sustaining the fall. The sperm samples obtained subsequently showed a rapid decline in quality. Frozen sperm from the first normal ejaculate obtained soon after the injury was used for assisted reproduction and has resulted in an ongoing pregnancy. CONCLUSION: This case highlights the importance of collection and freezing of seminal fluid within 2 weeks of spinal cord injury (SCI), or earlier to improve fertility outcomes in spinal cord-injured men, as the clinical stability of the patient may allow.

Second to fourth digit ratio (2D:4D) and testosterone in men.

AIM: To investigate the relationship between 2D:4D and testosterone in men attending an infertility clinic and men drawn from the general population. METHODS: Data on 2D:4D and testosterone from two samples were collected: (1) 43 men attending an infertility clinic, and (2) 51 men drawn from the general population without regard to fertility. RESULTS: In sample (1) there were negative associations between 2D:4D and testicular function, and men with lower 2D:4D in their right compared to left hand had higher testosterone levels than men with higher 2D:4D in their right compared to left hand. Sample (2) showed no significant associations between 2D:4D or side differences in 2D:4D and testosterone. CONCLUSION: Adult levels of testosterone may be related to aspects of 2D:4D in samples which contain men with compromised testicular function, but not in men from normative samples. Associations between 2D:4D and fertility-associated traits probably arise from early organisational effects of testosterone rather than from activational effects of current testosterone.

Expression of androgen receptors in upper human fetal reproductive tract.

BACKGROUND: Androgens play a key role in human fetal development. All androgens act through a single intracellular androgen receptor (AR), which is encoded by a single copy gene on the X chromosome. ARs are expressed as early as 9 weeks in the epithelium and mesenchyme of the urogenital sinus, paramesonephric (Müllerian) and mesonephric (Wolffian) ducts. METHODS: Using immunohistochemistry, we investigated the distribution of ARs in the gonads and lower genital tracts of 54 human fetuses at 8-11 weeks of gestation. Gender was determined by PCR. RESULTS: The AR was expressed in a similar pattern in both male and female fetuses. There appears to be no difference in expression in the mesonephros or the mesonephric ducts when male and female pelvises were compared. Expression in the female paramesonephric duct was within the epithelium, whereas, in the male pelvises, expression was in the mesenchyme of the paramesonephric duct. When AR expression was compared in the ovary and testes, both gonads seem to express AR at 9 weeks, but this expression was extended into the 10th week of gestation in the male. CONCLUSION: The specific pattern of AR expression implies a key role in gonadal development. However, the pattern of staining was similar in the gonads at 8 and 9 weeks in both sexes, although staining persisted longer in the testis until the 10th week. AR expression, therefore, is not a key determinant of human gonadal differentiation.

Cystic fibrosis mutation screening before assisted reproduction.

Cystic fibrosis (CF) is the most common autosomal recessive disease in Caucasians, with a reported incidence of one in 2500. In 97-98% of men with CF, a congenital bilateral absence of the vas deferens (CBAVD) blocks the transport of spermatozoa from testicular or epididymal structures to the outer genital tract, resulting in azoospermia. Abnormalities in semen parameters have also been identified in males with CF. Around 10% of obstructive azoospermia is congenital and is due to mutations the CF gene. It is therefore suggested that men who have CBAVD should be considered for CF screening prior to intracytoplasmic sperm injection (ICSI) treatment. Men with azoospermia of unknown origin should also be considered for screening. Couples requesting surgical sperm retrieval/ICSI should therefore be advised and counselled to bear an elevated risk for CF in their offspring. Counselling of these couples is vitally important in order to help them understand and cope with the diagnosis.

Current practice in the management of vasectomy reversal and unobstructive azoospermia in Merseyside & North Wales: a questionnaire-based survey.

OBJECTIVE: To investigate the current incidence of vasectomy reversal procedures, the techniques used and which practitioners use them. PATIENTS AND METHODS: Using a questionnaire, 130 general surgeons and urologists practising in Merseyside and North Wales were surveyed. RESULTS: The response rate was 74%, with 24 urological surgeons and 14 general surgeons undertaking vasectomy reversal. Annually, urological surgeons carried out significantly more procedures than did general surgeons, at 8.5 and 5.3 (P = 0.029), respectively. They were also more likely to use double-layer closure and microsurgical techniques, whilst significantly less likely to use stents. Urologists reported significantly greater patency rates, at 76% and 52% (P = 0.017), respectively, with no significant differences in subsequent pregnancy rates (30% vs 25%). Only one practitioner checked tubal patency in the female partner before vasectomy reversal. CONCLUSIONS: The use of vasectomy reversal is a cost-effective treatment for men wanting paternity after vasectomy. The technique used by the clinician and proper audit of the results require close attention; it would also appear to be obvious that all the partners of men seeking a vasectomy reversal should have their fertility status established before reversal, something that is clearly not done at present.

Endometrial integrin expression in women undergoing IVF and ICSI: a comparison of the two groups and fertile controls.

BACKGROUND: Integrins are thought to play a vital role in implantation. Three integrins in particular (alpha(4)beta(1), alpha(v)beta(3) and alpha(1)beta(1)) are all present during the implantation window. Defects in their expression have been linked to tubal disease, unexplained infertility and endometriosis. Hence, a reduced endometrial integrin expression would be expected in women attending for IVF due to these causes of infertility when compared with those with male factor infertility attending for ICSI. METHODS: Women attending for IVF (n = 25) and ICSI (n = 25) treatment were recruited, and timed endometrial biopsies were taken during the 'implantation window' (cycle day 20-24). A group of fertile women (n = 15) attending for sterilization was used as controls. RESULTS: There was no significant difference in integrin expression between patients undergoing IVF or ICSI. Neither did these groups differ from the control group. CONCLUSIONS: The endometrium in patients undergoing ICSI treatment is sometimes thought to be more receptive, as the infertility might be due to a male factor. This study shows that there is no significant difference in integrin expression between patients attending for IVF or ICSI and the control group. These data add to the increasing uncertainty about the clinical value of assessing the endometrium with only one marker, in this case integrins.

Surgical sperm retrieval after previous vasectomy and failed reversal: clinical implications for in vitro fertilization.

OBJECTIVE: To investigate the effect of the interval between previous vasectomy reversal on retrieval rates of epididymal and testicular spermatozoa using percutaneous epididymal sperm aspiration (PESA), or testicular sperm extraction (TESE), and the subsequent reproductive potential of these gametes in intracytoplasmic sperm injection (ICSI) cycles. PATIENTS AND METHODS: Sixty-six consecutive sperm retrievals were considered in patients who were azoospermic after previous vasectomy, of whom 54 had had a previous failed reversal, the remainder deciding against a reversal. PESA and TESE retrieval rates were noted, as were the time since vasectomy and the interval between vasectomy and unsuccessful reversal. The presence of palpable epididymal cysts was noted, with their effect on sperm retrieval rates. Fertilization and pregnancy rates were analysed in subsequent ICSI cycles using freshly retrieved spermatozoa or frozen-thawed cryopreserved spermatozoa. RESULTS: All 66 patients had sperm retrieved successfully; the success rates for PESA were not significantly affected by previous failed reversal when compared with patients who had not had a reversal, at 14 of 54 (26%) vs five of 12 (P=0.3). The interval since vasectomy did not affect PESA retrieval rates but there was a significantly poorer retrieval rate for PESA in the presence of palpable epididymal cysts, at seven of 35 (20%) vs 12 of 23 (52%) (P=0.012). Fertilization rates were significantly lower using cryopreserved spermatozoa retrieved from either the epididymis or testis (50% vs 70%, P=0.007), although subsequent implantation and pregnancy rates were not significantly different. CONCLUSION: Surgical sperm retrieval is successful in all cases of azoospermia secondary to vasectomy, either by PESA or TESE. There are no clinical markers to indicate which patients will have successful PESA after vasectomy, although the presence of epididymal cysts is associated with significantly lower retrieval rates. The reduction in fertilising ability of cryopreserved spermatozoa does not affect clinical pregnancy rates in ICSI cycles.

The effect of gonadotrophic stimulation on integrin expression in the endometrium.

BACKGROUND: Despite many recent advances in IVF treatment implantation rates per embryo transfer rarely exceed 30%. Three integrins (alpha(1)beta(1),alpha(4)beta(1) and alpha(v)beta(3)) have been shown to be expressed in the endometrium in a cyclically dependent manner and are thought therefore to play a vital role in the process of implantation. METHODS: The effect of gonadotrophin stimulation on the expression of these three integrins within the endometrium was investigated by examining biopsies from oocyte donation patients and comparing them with fertile controls. RESULTS: A delay in the maturation of the glandular epithelium was found in the oocyte donation patients. There was also a reduction in the expression of all three integrins in the glandular epithelium and also a reduced expression of the alpha(v)beta(3) integrin in the luminal epithelium. CONCLUSIONS: As these integrins have been shown to be important in implantation their reduced expression after IVF treatment may have an adverse effect on pregnancy rates.

Fructose concentrations in seminal plasma from men with nonobstructive azoospermia.

Fructose is secreted from the seminal vesicles and the accessory sex glands. It is the major carbohydrate source in seminal plasma and essential for normal sperm motility. The objective of this prospective descriptive study was to measure fructose concentrations in seminal plasma from men with obstructive and non-obstructive azoospermia, as determined by serum FSH, and to compare these with fructose concentrations in seminal plasma from men with normal semen parameters. A higher median fructose concentration was observed in seminal plasma from men with azoospermia and a raised serum FSH (3.22 mg/ml) compared to that from men with azoospermia and normal serum FSH (2.30 mg/ml) (p < 0.0001) and to that from normal controls (2.50 mg/ml) (p < 0.0002). This is the first report of increased seminal plasma concentrations of fructose in samples from men with non-obstructive azoospermia. This is either due to a decreased fructose metabolism associated with the azoospermia or, more likely, an increased fructose production from the seminal vesicles.

Comparison of ovarian response in right and left ovaries in IVF patients.

BACKGROUND: Anatomical and cyclical physiological differences exist between right and left ovaries which may affect their function and response to ovulation induction. Although authors have compared right and left ovarian response during IVF for patients with a unilateral diseased or absent ovary, no study has examined the response of normal ovaries to gonadotrophin stimulation within the same patient. We wished to determine if there were any significant differences between right and left ovarian response in patients with healthy ovaries having standard IVF treatment. METHODS: We performed a prospective observational case--controlled study in 200 consecutive IVF patients. The main outcome measures were the number of oocytes retrieved, fertilization rates, grade of embryos produced, pregnancy rates and live birth rates. RESULTS: Comparison of right versus left ovary revealed: number of oocytes 4.9 versus 4.7, percentage fertilization 61.3 versus 62.5%, percentage of grade 1 embryos 81 versus 83%, chemical pregnancy rate 33 versus 47% and live birth rate 27 versus 32% (all not significant). CONCLUSIONS: We conclude that there are no statistical differences between right and left ovarian response in IVF patients with healthy ovaries.

Peritoneal fluid concentrations of interleukin-4 in relation to the presence of endometriosis, its stage and the phase of the menstrual cycle.

BACKGROUND: Interleukin-4 (IL-4) is a cytokine with both stimulatory and inhibitory effects on the inflammatory system such as macrophage inhibition and T-cell activation. It is known to regulate several monocyte functions, including inhibition of the synthesis of cytokines such as IL-1, IL-6 and TNF-alpha as well as potentiating IL-8. METHOD: In an attempt to clarify the association between IL-4 and endometriosis, we measured the concentration of IL-4 in the peritoneal fluid of 52 women; 24 with endometriosis and 28 with no endometriosis, controlling for the phase of the cycle and the stage of disease. RESULTS: There was no difference in the concentrations of IL-4 between women with (n=28) and without endometriosis (n=24). No difference was found between the IL-4 concentrations in women with different stages of endometriosis. Levels of IL-4 did not show a difference according to the phase of the cycle in either group. CONCLUSION: Our results indicate no association between peritoneal fluid levels of IL-4 and endometriosis and hence suggest that IL-4 is not involved in the pathogenesis of endometriosis.

Peritoneal fluid concentrations of interleukin-11 in women with endometriosis.

OBJECTIVE: To determine the peritoneal fluid concentrations of interleukin-11 (IL-11) in women with endometriosis as compared with the control group. DESIGN: A prospective, controlled study. SETTING: The obstetrics and gynecology department of a teaching hospital and a university immunology department. PATIENT(S): Sixty consecutive women undergoing laparoscopic surgery for benign gynecological indications. INTERVENTION(S): Peritoneal fluid was obtained during laparoscopy, and the concentration of IL-11 was measured. MAIN OUTCOME MEASURE(S): Concentration of IL-11 in correlation with the presence of endometriosis, its stage, and the phase of the menstrual cycle. RESULT(S): IL-11 was detectable in the peritoneal fluid of 64% of women tested. Concentrations of IL-11 showed no correlation with the presence of endometriosis, the American Fertility Society stage of the disease, or the phase of the menstrual cycle. CONCLUSION(S): We found no evidence to suggest that IL-11 is involved in the pathogenesis of pelvic endometriosis.

Cystic fibrosis in infertility: screening before assisted reproduction: opinion.

Cystic fibrosis (CF) is the most common autosomal recessive disease in Caucasians. In 97-98% of men with CF, bilateral congenital absence of the vas deferens (CBAVD) blocks the transport of spermatozoa resulting in azoospermia. Abnormalities in sperm parameters have also been identified in males with CF. To date, over 800 disease-causing mutations of the CF transmembrane conductance regulator (CFTR) gene have been identified (also called ABCC7). Current legislation suggests that prior to intracytoplasmic sperm injection (ICSI) treatment, men with CBAVD or unexplained oligozoospermia should be considered for screening. If the male is negative with routine screening then the female partner is not screened. This is fundamentally wrong because if the female is screened and is found to be CF positive on routine testing, her partner would then need the fullest possible investigation of the CFTR gene. It is ideal to screen both partners in cases of oligozoospermia. However, if the resources are stretched, then only the female needs to be routinely screened because if she is negative, then the couple's residual risk of having a CF or CBAVD child will be reduced to 1:960. Only when the female is found to be a carrier does the male partner need routine screening followed by full testing for known mutations.

Follicular fluid concentrations of interleukin-12 and interleukin-8 in IVF cycles.

OBJECTIVE: To investigate the role of interleukin-12 (IL-12) and IL-8 in the periovulatory follicular fluid during in vitro fertilization cycles. DESIGN: A prospective study. SETTING: Reproductive Medicine Unit, Liverpool Women's Hospital, United Kingdom. PATIENT(S): Women undergoing in vitro fertilization treatment. INTERVENTION(S): IL-8 and IL-12 concentrations in follicular fluid samples that had been collected during transvaginal oocyte retrieval were measured using an enzyme-linked immunosorbent assay (ELISA). Cytokine concentrations were correlated to fertilization rates and treatment outcome. MAIN OUTCOME MEASURE(S): Fertilization rates and ultrasonographic evidence of intrauterine pregnancy by 4 weeks after embryo transfer. RESULT(S): Failed fertilization in women with detectable IL-12 was significantly higher (45.5%) than in the IL-12 negative group (6.1%), P=.01. None of the women with detectable IL-12 achieved a pregnancy at the end of the treatment (P=.01). IL-8 was present in the follicular fluid of all women, and no difference in its concentrations was found between the pregnant and nonpregnant groups. No correlation was found between the follicular fluid concentrations of IL-8 and fertilization rates. CONCLUSION(S): The presence of IL-12 in the follicular fluid appears to be associated with a negative outcome in IVF treatment. Interleukin-8 appears to be an essential part of folliculogenesis, although its concentration is not associated with fertilization or implantation rates.

Role of mitotic control in spermatogenesis.

OBJECTIVE: To study the correlation between the incidence of sex chromosome aneuploidies in the somatic cells and spermatozoa in karyotypically normal infertile men and fertile donors. DESIGN: A prospective, phase two, controlled study. SETTING: A teaching Hospital Reproductive Medicine and Medical Genetics Units. PATIENT(S): Ten patients with idiopathic oligozoospermia and 10 sperm donors with proven fertility, all with a normal karyotype 46, XY. INTERVENTION(S): Multicolor fluorescence in situ hybridization (FISH) of peripheral blood lymphocytes and spermatozoa using a probe cocktail containing the alpha satellite DXZ1 for the X centromere, DYZ1 for the heterochromatic region of the long arm of the Y, and cosmids D21S259, D21S341, and D21S342 for Down syndrome critical region of chromosome 21. MAIN OUTCOME MEASURE(S): The incidence of chromosome X, Y, and 21 aneuploidies in peripheral lymphocytes and spermatozoa in both groups. RESULT(S): The incidence of aneuploidies related to chromosomes X, Y, and 21 were significantly higher in peripheral lymphocytes and spermatozoa of infertile men compared with donors. There was a positive correlation between the incidence of chromosome aneuploidies in the somatic cells and sperm in all men. CONCLUSION(S): These findings provide suggestive evidence for the importance of mitosis in spermatogenesis and the role of mitotic instability in unexplained oligozoospermia.

Assessment of testicular core temperatures using microwave thermography.

A new method for the reliable assessment of testicular core temperature using microwave technology is presented. This study was designed to assess whether alterations in testicular thermoregulation could be reliably demonstrated in patients with clinically apparent varicoceles (n = 36), in those with idiopathic male infertility (n = 52) and in fertile donors (n = 20) using this new microwave thermographic technique. The measurements obtained were found to be reliable and reproducible. Testicular core temperature measurements were significantly different between the groups (P < 0.001). Furthermore, there was a temperature gradient between the scrotal neck and the testicular core in all groups; testicular core temperatures were lower than scrotal neck temperatures. The magnitude of this temperature difference was also significantly different (P < 0.001) between the groups. Microwave testicular thermography is a new technique that is safe and accurate. Preliminary results suggest altered testicular thermoregulation in a group of patients with impaired spermatogenesis with and without varicocele. Testicular temperature profiles obtained by microwave thermography may be of value in the assessment of infertile men with or without a varicocele.