Probing human sperm metabolism using 13C-magnetic resonance spectroscopy.

STUDY QUESTION: Can 13C-Magnetic Resonance Spectroscopy (MRS) of selected metabolites provide useful information about human sperm metabolism and how glycolysis or oxidative phosphorylation are used by different sperm populations? SUMMARY ANSWER: Sperm populations, prepared by density gradient centrifugation (DGC) and incubated with either 13Cu-glucose, 13Cu-fructose or 13C1-pyruvate, showed consistent evidence of metabolism generating principally lactate and more intermittently bicarbonate, and significantly more lactate was produced from 13Cu-glucose by vital or motile sperm recovered from the 40/80% interface compared to those from the pellet, which could not be accounted for by differences in the non-sperm cells present. WHAT IS KNOWN ALREADY: Previous studies have focused on CO2 or other specific metabolite production by human sperm and there remains considerable debate about whether glycolysis and/or oxidative phosphorylation is the more important pathway for ATP production in sperm. STUDY DESIGN, SIZE, DURATION: Sperm populations were prepared by DGC and subjected to 13C-MRS to answer the following questions. (i) Is it possible to detect human sperm metabolism of 13C substrates implicated in energy generation? (ii) What are the kinetics of such reactions? (iii) Do different sperm populations (e.g. '80%' pellet sperm and '40%' interface sperm) utilise substrates in the same way? Semen samples from 97 men were used in these experiments; 52 were used in parallel for aims (i) and (ii) and 45 were used for aim (iii). PARTICIPANTS/MATERIALS, SETTING, METHODS: Sperm populations were prepared from ejaculates of healthy men using a Percoll/Phosphate Buffered Saline (PBS) DGC and then incubated with a range of 13C-labelled substrates (13Cu-glucose, 13Cu-fructose, 13C1-pyruvate, 13C1-butyrate, 13C3-lactate, 13C2,4-D-3-hydroxybutyrate, 13C5-l-glutamate, 13C1,2-glycine or 13Cu-galactose) along with penicillin/streptomycin antibiotic at 37°C for 4 h, 24 h or over 48 h for an estimated rate constant. Sperm concentration, vitality and motility were measured and, for a subset of experiments, non-sperm cell concentration was determined. A 9.4 T magnetic resonance spectrometer was used to acquire 1D 13C, inverse gated 1H decoupled, MRS spectra. Spectrum processing was carried out using spectrometer software and Matlab scripts to determine peak integrals for each spectrum. MAIN RESULTS AND THE ROLE OF CHANCE: 13Cu-glucose, 13Cu-fructose and 13C1-pyruvate were consistently converted into lactate and, to a lesser extent, bicarbonate. There was a significant correlation between sperm concentration and lactate peak size for 13Cu-glucose and 13Cu-fructose, which was not observed for 13C1-pyruvate. The lactate peak did not correlate with the non-sperm cell concentration up to 6.9 × 106/ml. The concentration of 13Cu-glucose, 13Cu-fructose or 13C1-pyruvate (1.8, 3.6, 7.2 or 14.4 mM) had no influence on the size of the observed lactate peak over a 4 h incubation. The rate of conversion of 13C1-pyruvate to lactate was approximately three times faster than for 13Cu-glucose or 13Cu-fructose which were not significantly different from each other. After incubating for 4 h, the utilisation of 13Cu-glucose, 13Cu-fructose or 13C1-pyruvate by sperm from the '40%' interface of the DGC was no different from those from the pellet when normalised to total sperm concentration. However, after normalising by either the vital or motile sperm concentration, there was a significant increase in conversion of 13Cu-glucose to lactate by '40%' interface sperm compared to pellet sperm (Vital = 3.3 ± 0.30 × 106 vs 2.0 ± 0.21 × 106; P = 0.0049; Motile = 7.0 ± 0.75 × 106 vs 4.8 ± 0.13 × 106; P = 0.0032. Mann-Whitney test P < 0.0055 taken as statistically significant). No significant differences were observed for 13Cu-fructose or 13C1-pyruvate. LARGE SCALE DATA: Not applicable. LIMITATIONS, REASONS FOR CAUTION: Only 13C labelled metabolites that accumulate to a sufficiently high concentration can be observed by 13C MRS. For this reason, intermediary molecules in the metabolic chain are difficult to observe without trapping the molecule at a particular step using inhibitors. Non-sperm cell concentration was typical of the general population and no link was found between these cells and the magnitude of the 13C-lactate peak. However, it is possible that higher concentrations than the maximum observed (6.9 × 106/ml) may contribute to exogenous substrate metabolism in other experiments. WIDER IMPLICATIONS OF THE FINDINGS: 13C-MRS can provide information on the underlying metabolism of multiple pathways in live sperm. Dysfunction in sperm metabolism, as a result of either impaired enzymes of lack of metabolisable substrate, could be detected in sperm by a non-destructive assay, potentially offering new treatment options to improve overall sperm quality and outcomes for reproduction. STUDY FUNDING AND COMPETING INTERESTS: This work was supported by the Medical Research Council Grant MR/M010473/1. The authors declare no conflicts of interest.

Effects of Chlamydia trachomatis infection on sperm chromatin condensation and DNA integrity.

The present study was performed to investigate the relation of Chlamydia trachomatis infection to sperm chromatin/DNA integrity in a population of infertile men (male partner of infertile couples) from Iran. Blood, semen and first-void urine samples were obtained from 250 infertile men. Data were analysed with regard to the results of (i) serological analysis for specific antibodies to C. trachomatis in serum; (ii) the presence of C. trachomatis and DNA in first-void urine; and (iii) in the semen sample of the male partner, in addition to sperm analysis, four different tests (aniline blue, chromomycin A3, acridine orange and TUNEL) were used to detect sperm chromatin and DNA abnormalities. The main conclusions of the results were: (i) no evidence of C. trachomatis infection in semen samples was found; (ii) sperm DNA fragmentation and chromatin studies were not correlated with C. trachomatis diagnosis; (iii) the percentage of DNA fragmentation is positively correlated with the percentage of immotile sperm but negatively with semen volume, normal morphology; and (iv) in sperm chromatin evaluations, only the percentage of chromatin protamination was related to male age.

Classification systems in Gestational trophoblastic neoplasia - Sentiment or evidenced based?

The classification system for Gestational trophoblastic neoplasia (GTN) has proved a controversial topic for over 100years. Numerous systems simultaneously existed in different countries, with three main rival classifications gaining popularity, namely histological, anatomical and clinical prognostic systems. Until 2000, prior to the combination of the FIGO and WHO classifications, there was no worldwide consensus on the optimal classification system, largely due to a lack of high quality data proving the merit of one system over another. Remarkably, a validated, prospectively tested classification system is yet to be conducted. Over time, increasing criticisms have emerged regarding the currently adopted combined FIGO/WHO classification system, and its ability to identify patients most likely to develop primary chemotherapy resistance or disease relapse. This is particularly pertinent for patients with low-risk disease, whereby one in three patients are resistant to first line therapy, rising to four out of five women who score 5 or 6. This review aims to examine the historical basis of the GTN classification systems and critically appraise the evidence on which they were based. This culminates in a critique of the current FIGO/WHO prognostic system and discussion surrounding clinical preference versus evidence based practice.

1H Magnetic Resonance Spectroscopy of live human sperm.

STUDY QUESTION: Can 1H Magnetic Resonance Spectroscopy (MRS) be used to obtain information about the molecules and metabolites in live human spermatozoa? SUMMARY ANSWER: Percoll-based density gradient centrifugation (DGC) followed by a further two washing steps, yielded enough sperm with minimal contamination (<0.01%) from seminal fluid to permit effective MRS which detected significant differences (P < 0.05) in the choline/glycerophosphocholine (GPC), lipid and lactate regions of the 1H MRS spectrum between sperm in the pellet and those from the 40%/80% interface. WHAT IS KNOWN ALREADY: Current methods to examine sperm are either limited in their value (e.g. semen analysis) or are destructive (e.g. immunohistochemistry, sperm DNA testing). A few studies have previously used MRS to examine sperm, but these have either looked at seminal plasma from men with different ejaculate qualities or at the molecules present in pooled samples of lyophilized sperm. STUDY DESIGN, SAMPLES/MATERIALS, METHODS: Sperm suspended in phosphate buffered saline (PBS) at 37°C were examined by 1H MRS scanning using a 1H excitation-sculpting solvent suppression sequence after recovery from fresh ejaculates by one of three different methods: (i) simple centrifugation; (ii) DGC with one wash; or (iii) DGC with two washes. In the case of DGC, sperm were collected both from the pellet ('80%' sperm) and the 40/80 interface ('40%' sperm). Spectrum processing was carried out using custom Matlab scripts to determine; the degree of seminal plasma/Percoll contamination, the minimum sperm concentration for 1H MRS detection and differences between the 1H MRS spectra of '40%' and '80%' sperm. MAIN RESULTS AND THE ROLE OF CHANCE: DGC with two washes minimized the 1H MRS peak intensity for both seminal plasma and Percoll/PBS solution contamination while retaining sperm specific peaks. For the MRS scanner used in this study, the minimum sperm concentration required to produce a choline/GPC 1H MRS peak greater than 3:1 signal to noise ratio (SNR) was estimated at ~3 × 106/ml. The choline/GPC and lactate/lipid regions of the 1H spectrum were significantly different by two-way ANOVA analysis (P < 0.0001; n = 20). ROC curve analysis of these region showed significant ability to distinguish between the two sperm populations: choline/GPC ROC AUC = 0.65-0.67, lactate/lipid ROC AUC = 0.86-0.87. LIMITATIONS, REASONS FOR CAUTION: Only 3-4 semen samples were used to assess the efficacy of each sperm washing protocol that were examined. The estimated minimum sperm concentration required for MRS is specific to the hardware used in our study and may be different in other spectrometers. Spectrum binning is a low resolution analysis method that sums MRS peaks within a chemical shift range. This can obscure the identity of which metabolite(s) are responsible for differences between sperm populations. Further work is required to determine the relative contribution of somatic cells to the MRS spectrum from the '40%' and '80%' sperm. WIDER IMPLICATIONS OF THE FINDINGS: 1H MRS can provide information about the molecules present in live human sperm and may therefore permit the study of the underlying functional biology or metabolomics of live sperm. Given the relatively low concentration of sperm required to obtain a suitable MRS signal (~3 × 106/ml), this could be carried out on sperm from men with oligo-, astheno- or teratozoospermia. This may lead to the development of new diagnostic tests or ultimately novel treatments for male factor infertility. STUDY FUNDING AND COMPETING INTEREST(S): This work was supported by the Medical Research Council Grant MR/M010473/1. The authors declare no conflicts of interest.

Relationship between Chlamydia trachomatis and Mycoplasma genitalium infection and pregnancy rate and outcome in Iranian infertile couples.

The study was performed to investigate the prevalence of Chlamydia trachomatis and Mycoplasma genitalium in a population of infertile couples from Iran and how this relates to tubal factor infertility, pregnancy rate and outcome of pregnancy. Blood, semen and first-void urine samples were obtained from 250 infertile couples and 250 fertile women as a control. Infertile couples were followed up after 24 months to determine diagnosis, referral for assisted conception, any pregnancy and pregnancy outcome. Data were analysed with regard to the results of (i) serological analysis for specific antibodies to C. trachomatis in serum; (ii) the presence of C. trachomatis and M. genitaliumDNA in first-void urine; and (iii) in a semen sample of the male partner. Prevalence of C. trachomatis in our study population was comparable to other studies using similar methods and test specimens. No evidence of M. genitalium infection was found. Detection of C. trachomatis in one partner rarely correlated with infection in the other. The risk of tubal factor infertility and the probability of pregnancy and pregnancy outcome were unrelated to the results of serological tests for C. trachomatis antibodies or the presence of C. trachomatisDNA in first-void urine of both partners and in a semen sample provided by the male.

Semen inflammatory markers and Chlamydia trachomatis infection in male partners of infertile couples.

Previous studies have given conflicting results about the effect of generally infection and Chlamydia trachomatis on seminal ILs and semen parameters. The aim of this study was to investigate the relationship between semen quality and the level of seminal interleukins (ILs) in infertile couples with C. trachomatis. Blood, first void urine (FVU) and semen were obtained from 250 infertile men who had failed to conceive after 12 months of trying. Serological analysis for specific IgA, IgM and IgG antibodies to C. trachomatis in serum, the presence of C. trachomatis in FVU and semen sample and semen analysis were carried out. The main results are as follows: (i) elevated IL-6 and IL-8 are observed in C. trachomatis-positive men, but this is not significant and it varies by diagnostic method; and (ii) IL-6 and IL-8 levels were correlated with each other and the concentration of leucocytes, but IL-8 was correlated with semen volume and patient's age. This study showed that men with such an infection in FVU samples (PCR positive) had only lower semen volume compared with men without infection.

Modifiable and non-modifiable risk factors for poor sperm morphology.

STUDY QUESTION: Are common lifestyle factors associated with poor sperm morphology? SUMMARY ANSWER: Common lifestyle choices make little contribution to the risk of poor sperm morphology. WHAT IS KNOWN ALREADY: Although many studies have claimed that men's lifestyle can affect sperm morphology, the evidence is weak with studies often underpowered and poorly controlled. STUDY DESIGN, SIZE, DURATION: Unmatched case-referent study with 318 cases and 1652 referents. Cases had poor sperm morphology (<4% normal forms based on 200 sperm assessed). Exposures included self-reported exposures to alcohol, tobacco, recreational drugs as well as occupational and other factors. PARTICIPANTS/MATERIALS, SETTING, METHODS: Eligible men, aged 18 years or above, were part of a couple who had been attempting conception without success following at least 12 months of unprotected intercourse and also had no knowledge of any semen analysis before being enrolled. They were recruited from 14 fertility clinics across the UK during a 37-month period from 1 January 1999. MAIN RESULTS AND THE ROLE OF CHANCE: Risk factors for poor sperm morphology, after adjustment for centre and other risk factors, included: (i) sample production in summer [odds ratio (OR) = 1.99, 95% confidence interval (CI) 1.43-2.72]; and (ii) use of cannabis in the 3 months prior to sample collection in men aged ≤30 years (OR = 1.94, 95% CI 1.05-3.60). Men who produced a sample after 6 days abstinence were less likely to be a case (OR = 0.64, 95% CI 0.43-0.95). No significant association was found with body mass index, type of underwear, smoking or alcohol consumption or having a history of mumps. This suggests that an individual's lifestyle has very little impact on sperm morphology and that delaying assisted conception to make changes to lifestyle is unlikely to enhance conception. LIMITATIONS, REASONS FOR CAUTION: Data were collected blind to outcome and so exposure information should not have been subject to reporting bias. Less than half the men attending the various clinics met the study eligibility criteria and among those who did, two out of five did not participate. It is not known whether any of those who refused to take part did so because they had a lifestyle which they did not want subjected to investigation. Although the power of the study was sufficient to draw conclusions about common lifestyle choices, this is not the case for exposures that were rare or poorly reported. WIDER IMPLICATIONS OF THE FINDINGS: All participating clinics saw patients at no cost (under the UK National Health Service) and the study population may differ from those in countries without such provision. Even within the UK, low-income couples may choose not to undertake any investigation believing that they would subsequently be unable to afford treatment. Since a computer performed the measurements of sperm morphology, these results may not be comparable with studies where sperm morphology was assessed by other methods. STUDY FUNDING/COMPETING INTERESTS: The study was funded by the UK Health and Safety Executive, the UK Department of Environment, Transport and the Regions, the UK Department of Health (Grant Code DoH 1216760) and the European Chemical Industry Council (grant code EMSG19). No competing interests declared.

Sperm protection in the male reproductive tract by Toll-like receptors.

Sperm function can be affected by infection. Our understanding of innate immune system molecular mechanisms has been expanded, by the discovery of 'Toll-like receptors' (TLRs). It seems that these receptors could play a critical role in the protection of spermatozoa. This study seeks to examine the presence and distribution of TLRs in different parts of the human male reproductive tract and spermatozoa. So, TLR gene expression was examined by RT-PCR. Quantitative real-time PCR (Q-PCR) analysis used to compare the expression of TLRs in all sections of the male reproductive tract and TLRs 2, 3 and 4 in testicular sperm extraction (TESE) samples, which contained spermatozoa (TESE+) and those that did not (TESE-). Results showed that all TLR genes were expressed in different parts of the human male reproductive tract and spermatozoa. Moreover, Q-PCR indicated that the relative expression of TLRs did not significantly change in different parts of the male reproductive tract but this technique has shown only relative TLR2 expression in TESE- is lower than TESE+ samples. It could be concluded that TLRs may provide a broad spectrum of protection from infection in the male reproductive tract. Furthermore, TLRs may influence on the developmental process during spermatogenesis.

Discussing fertility preservation at the time of cancer diagnosis: dissatisfaction of young females.

BACKGROUND: Sperm banking is widely available for post-pubertal male cancer patients but options for females remain limited. Anecdotal evidence suggests that fertility issues may be inadequately discussed with females. To understand the experience of both sexes in the UK, surveys of young cancer survivors were performed 7 years apart. PROCEDURE: Data were collected from young cancer survivors aged over 13 years at diagnosis, attending support group conferences held in 2004 and 2011. Data were collected anonymously using remote handsets in response to questions projected on the screen during plenary sessions. RESULTS: A total of 81 female and 69 males responded in 2004, and 69 females and 71 males in 2011. In both years, most males reported fertility discussions taking place before treatment started and they were generally satisfied with it. However in both years, fewer females recall a discussion about fertility and they were generally less satisfied. Although in 2011 more females reported a fertility discussion prior to the beginning of treatment, they were no more satisfied than the females in 2004 whose fertility discussion were more likely to take place after treatment had started. CONCLUSIONS: Whilst male cancer survivors in the UK are generally satisfied about the frequency and timing of discussions about fertility, females are not. Although in 2011 fertility discussions with females more often took place before treatment began, they were no more satisfied than females in 2004. This may reflect the approach by professionals or the absence of effective fertility preservation strategies for them.

Monitoring fertility (semen analysis) by cancer survivors who banked sperm prior to cancer treatment.

STUDY QUESTION: What medical and psychological variables predict why men with banked sperm do not return for semen analysis after their cancer treatment has ended? SUMMARY ANSWER: Men who decline the offer of semen analysis are less likely to have reported adverse side effects during cancer treatment, and have a more negative experience of banking sperm and a more negative attitude towards disposal of their stored semen than those who attend. WHAT IS KNOWN ALREADY: Previous authors have noted that male cancer survivors seem reluctant to have their fertility tested after their treatment has ended. Moreover, the utilization rates of banked sperm are very low (<10%) and the majority of samples are kept for many years without being used. STUDY DESIGN, SIZE AND DURATION: A cross-sectional study of 499 cancer survivors who were sent a questionnaire about their views on sperm banking, fertility and post-treatment semen analysis between April 2008 and December 2010. PARTICIPANTS AND SETTING: Men (aged 18-55 years) who had banked sperm in Sheffield and Nottingham (UK) prior to gonadotoxic treatment for cancer more than 5 years previously. MAIN RESULTS AND THE ROLE OF CHANCE: Completed questionnaires were received from 193 men (38.7% response rate) whose samples had been banked for 9.18 ± 3.70 years (range = 4.94-26.21) and whose current age was 35.08 ± 7.08 years (range = 21.58-54.34; mean ± SD). One-third (35.8%) had never attended for semen analysis. In multivariate analysis, the odds of not attending for semen analysis were significantly greater among men who did not experience adverse treatment side effects [odds ratio (OR) = 5.72, 95% confidence interval (CI) = 2.10-15.56], who reported a more negative experience of banking sperm (OR = 1.82, 95% CI = 1.17-2.82) and a more negative attitude to disposal of their stored semen (OR = 1.56, 95% CI = 1.01-2.42). LIMITATIONS AND REASONS FOR CAUTION: Only 38.7% of those eligible agreed to take part. We do not know the characteristics of men who declined to take part, if they agreed to attend semen analysis without completing the questionnaire or whether they had chosen to have semen analysis performed elsewhere (e.g. private sector). Some of the measures used (e.g. experience of banking sperm) relied on men's recall of events many years previously. WIDER IMPLICATIONS OF THE FINDINGS: New strategies are required to encourage these men to engage with fertility monitoring programmes if sperm banks are to be used cost-effectively and men are to be given appropriate fertility advice. STUDY FUNDING AND COMPETING INTERESTS: This paper was supported by funding from Cancer Research-UK to C.E., A.A.P. and R.R. (C481/A8141). The views expressed are those of the authors. No competing interests declared.

Finding out about sperm banking: what information is available online for men diagnosed with cancer?

Sperm banking is routinely offered to men where there is a risk of infertility following cancer treatment but uptake is lower than expected. Since these men may turn to the internet for information, we used the search engine www.google.com to identify the material available about sperm banking and fertility preservation options. Sixty-six resources (NHS/Private Clinic, Charity, Press Releases, General and Forums/Blogs) fulfilled the criteria for inclusion and were examined for quality including readability, layout and content. The most frequently reported information related to: (1) effects of cancer treatment on fertility (77.3%); (2) reasons to bank sperm (69.7%); and (3) fertility recovery after treatment (57.6%). Information about maintaining contact with the sperm bank (18.2%) and disposal of banked samples (10.6%) was less often included. The quality of information available on the Internet about sperm banking was variable. The readability of all resources was assessed as 'fairly difficult', i.e. reading skills required were too complex for the average member of the public to understand. Furthermore, visual presentation of material (e.g. lay out) did not facilitate easy reading. More attention should be given to information about longer-term issues, such as fertility recovery and the use or disposal of banked sperm.

Modifiable and non-modifiable risk factors for poor semen quality: a case-referent study.

STUDY QUESTION: Are common lifestyle factors associated with low-motile sperm concentration (MSC)? SUMMARY ANSWER: Common lifestyle choices make little contribution to the risk of low MSC. WHAT IS KNOWN AND WHAT THIS PAPER ADDS: Reviews of male subfertility often highlight how aspects of men's adult lifestyle can significantly increase their risk of subfertility but the strength of supporting evidence is weak. In this study, although low MSC was associated with a history of testicular surgery, being in manual work, not wearing loose underwear and black ethnicity, no relation was found to consumption of alcohol, use of tobacco or recreational drugs or high body mass index (BMI). These results suggest that delaying assisted conception to make changes to lifestyle is unlikely to enhance conception. DESIGN: Unmatched case-referent study with 939 cases and 1310 referents. Cases had a low-MSC relative to the time since last ejaculation (<12 × 10(6) for 3 days of abstinence). Exposures included self-reported exposures to alcohol, tobacco, recreational drugs as well as occupational and other factors. PARTICIPANTS AND SETTING: Eligible men, aged 18 or above, were part of a couple who had been attempting conception without success following at least 12 months of unprotected intercourse and also had no knowledge of any semen analysis. They were recruited from 14 fertility clinics across the UK during a 37-month period from 1 January 1999. MAIN RESULTS AND THE ROLE OF CHANCE: Risk factors for low MSC, after adjustment for centre and confounding factors, included a history of testicular surgery [odds ratio = 2.39, 95% confidence interval (CI): 1.75, 3.28], being in manual work [odds ratio (OR) = 1.28, 95% CI: 1.07, 1.53] or not working (OR = 1.78, 95% CI: 1.22, 2.59) and having black ethnicity (OR = 1.99, 95% CI: 1.10, 3.63). Conversely, men who wore boxer shorts (OR = 0.76, 95% CI: 0.64, 0.92) or who had a previous conception (OR = 0.71, 95% CI: 0.60, 0.85) were less likely to be a case. No significant association was found with smoking and alcohol consumption, the use of recreational drugs, a high BMI or having a history of mumps or fever. BIAS, CONFOUNDING AND OTHER REASONS FOR CAUTION: Data were collected blind to outcome, and exposure information should not have been subject to reporting bias. Among men attending the various clinics less than half met the study eligibility criteria and among those who did, two out of five were not recruited. It is not known whether any of those who refused to take part did so because they had a lifestyle they did not want subjected to investigation. Although the power of the study was sufficient to draw conclusions about common lifestyle choices, it cannot comment on exposures that are perhaps rare and poorly reported: the finding that use of street drugs was unrelated to low MSC cannot be assumed to apply to all such drugs and all patterns of use. The case definition did not consider sperm morphology or sperm DNA integrity. GENERALIZABILITY TO OTHER POPULATIONS: All participating clinics saw patients at no cost (under the UK National Health Service) and the study population may differ from those in countries without such provision. Even within the UK, low-income couples may choose not to undertake any investigation believing that they would subsequently be unable to afford treatment.

The legacy of sperm banking: how fertility monitoring and disposal of sperm are linked with views of cancer treatment.

BACKGROUND: Sperm banking is recommended for all men before cancer treatment, which carries a risk of long-term gonadal damage. However, relatively few men take up the offer. Among them, few attend for fertility monitoring or agree to sperm disposal where fertility recovers. Sperm banks are therefore burdened by long-term storage of samples that may not be needed for conception, with implications for healthcare resources. The aims here were to determine the views of men regarding personal benefits of sperm banking, and the advantages and disadvantages of fertility monitoring and disposal in the longer term. METHODS: Semi-structured interviews were conducted with 19 men who were diagnosed with cancer and had banked sperm at least 5 years previously. Men were asked to recall their experiences from diagnosis to the present time, focusing on the consequences for their fertility. Interviews were transcribed and analysed using Interpretative Phenomenological Analysis. RESULTS: Results are discussed in relation to decisions surrounding banking sperm, fertility monitoring and attitudes to disposal of banked sperm. Complex attitudes were identified, with men's views reflecting their understanding of their current and future fertility and the possible trajectory of cancer itself. Men are overwhelmed by information on diagnosis and fail to understand the implications of cancer treatment for their future fertility. CONCLUSIONS: On diagnosis, men are given large amounts of information about cancer and treatment but fail to understand the longer-term implications of sperm banking. These implications need to be specifically addressed at subsequent appointments in order to optimize fertility monitoring and timely disposal of sperm samples.

Crossing borders for fertility treatment: motivations, destinations and outcomes of UK fertility travellers.

BACKGROUND: There are few systematic studies of the incidence of cross-border fertility care and even fewer reports of qualitative research with those undertaking treatment outside their country of origin. This paper reports findings from a qualitative study of UK residents with experience of cross-border care: the socio-demographic characteristics of UK travellers; their reasons for seeking treatment abroad; the treatments they sought; the destinations they chose and the outcomes of their treatment. METHODS: Data regarding cross-border fertility treatment were collected from a purposive sample of 51 people by means of in-depth, semi-structured interviews between May 2009 and June 2010. Data were analysed using a systematic thematic coding method and also subjected to quantitative translation. RESULTS: Patient motivations for travelling abroad are complex. A desire for timely and affordable treatment with donor gametes was evident in a high number of cases (71%). However, most people gave several reasons, including: the cost of UK treatment; higher success rates abroad; treatment in a less stressful environment and dissatisfaction with UK treatment. People travelled to 13 different countries, the most popular being Spain and the Czech Republic. Most organized their own treatment and travel. The mean age of women seeking treatment was 38.8 years (range 29-46 years) and the multiple pregnancy rate was 19%. CONCLUSIONS: UK residents have diverse reasons for, and approaches to, seeking overseas treatment and do not conform to media stereotypes. Further research is needed to explore implications of cross-border treatment for donors, offspring and healthcare systems.

The value of testing semen for Chlamydia trachomatis in men of infertile couples.

Chlamydia trachomatis is an important bacterial cause of infertility. In men, the recommended specimen for diagnosing chlamydial infection is urine, with little or no emphasis placed on testing semen. A systematic review of the literature was conducted to search for studies in which men undergoing investigations for infertility produced both samples of urine and semen that were tested concurrently for C. trachomatis. An analysis of these studies was then performed. From 322 papers identified from the US National Library of Medicine PubMed database, 14 were selected for a detailed review and 11 were analysed further. Overall, the size of the study groups was only modest and differences between the studies included variation in geography and test methodology, especially whether commercial testing systems had been used. There was also a lack of consistency with regard to including men with azoospermia. Patients were typically 30-35 years old and the median duration of infertility was about 4 years. Of those patients positive for C. trachomatis in the 11 studies, 56% could be detected in both semen and urine, 20% only in urine and 23% only in semen. Deficiencies in existing studies would not allow for a meta-analysis, emphasizing the need for further research in this area for which a number of recommendations are made.

Environmental and lifestyle factors associated with sperm DNA damage.

Semen quality in the adult male can be affected by a number of environmental and lifestyle factors. However, most studies to date have only considered how sperm concentration, motility and morphology are affected by such exposures. More recent studies, outlined in this review, have begun to examine the effect of environmental and lifestyle factors on sperm DNA integrity. Important factors appear to include: (i) Physical agents such as radiation and heat; (ii) Chemical agents such as cigarette smoke, airborne pollutants, and chemotherapeutic drugs; and (iii) Biological factors including sexually transmitted infections, increasing male age, elevated body mass index and medical conditions such as insulin dependent diabetes. A number of hypotheses have been proposed to account for how sperm DNA might be damaged by these factors. There is, however, uncertainty about the most appropriate laboratory test(s) to identify and quantify such DNA damage and no convincing evidence on possible therapeutic measures.

Mannose-binding lectin is present in human semen and modulates cellular adhesion of Neisseria gonorrhoeae in vitro.

Mannose-binding lectin (MBL) is an innate immune molecule present in blood and some mucosal tissues, which can influence microbial attachment and inflammatory responses of host cells during infection. In this study MBL was found to be present at a low concentration in semen samples in the range 1.2-24.9 ng/ml. Co-incubation of bacteria with semen resulted in the binding of MBL to the bacterial surface. Neisseria gonorrhoeae is a common cause of genitourinary infection. MBL bound to N. gonorrhoeae with strain-to-strain variation in the intensity of binding and nature of the bacterial receptor. Pretreatment with MBL concentrations similar to those found in human serum modulated the adhesion of N. gonorrhoeae strain FA1090 but not strain MS11 to epithelial cells. This effect was dose-dependent. This work demonstrates that MBL is present in human semen and modifies cellular responses to N. gonorrhoeae in a concentration-dependent manner.

Fertility issues in survivors from adolescent cancers.

Infertility is a common and distressing late-effect of cancer treatment. Whist sperm banking for post-pubertal males and embryo freezing for women (who are in a stable relationship at the time of treatment) are highly successful fertility preservation strategies, for females without a partner (including young and pre-pubescent girls) and pre-pubescent boys (or azoospermic men), there remain no effective approaches. Whilst the biological effects of cancer treatments on the reproductive system are well described, there are few data on the relative incidence of infertility (failure to conceive after one year of trying) in cancer survivors. This makes it difficult to advise survivors about their future fertility prospects. Whilst some will undoubtedly conceive naturally with their partner, others will require assisted conception treatment of which in vitro fertilisation (IVF) and intra-cytoplasmic sperm injection (ICSI) are the most common. Pregnancy outcomes of cancer survivors are generally good, although there is increased risk of pre-term birth and low birth-weight in the offspring of women who have received pelvic irradiation. There is no increased incidence of genetic disease or cancer incidence in the offspring of cancer survivors. Current research directions are focussing on alternative fertility preservation strategies including in vitro maturation techniques, xenotransplantation and the development of technology to create artificial gametes in the laboratory. Finally, although the reproductive techniques discussed are highly effective, country specific differences in the legal framework means that cancer survivors may be denied access to certain treatments (e.g. embryo cryopreservation) because they are forbidden by specific national legislation.

Relationship between the fertile period and sperm transport in the bitch.

Following ejaculation into the vagina, dog sperm remain functionally competent for many days, acquire the ability to fertilize, and are delivered to an appropriate site within the uterine tube synchronously with the appearance of fertile oocytes. The mechanisms involved in regulating this system are complex, and allow for sperm storage within the female reproductive tract. The aim of this paper is to review current knowledge of the transportation of sperm and their biology within the reproductive tract of the bitch.