Spermatogenesis in Hodgkin's lymphoma patients: a retrospective study of semen quality before and after different chemotherapy regimens.

STUDY QUESTION: Is spermatogenesis impairment caused by Hodgkin's lymphoma (HL) itself or by the various treatments? SUMMARY ANSWER: HL is not itself the main cause of impaired spermatogenesis, which is instead affected by the treatment; the extent of impairment depends on the type of treatment and the number of cycles. WHAT IS KNOWN ALREADY: Data in the literature are contradictory, although most studies found poor semen quality in HL patients prior to treatment. The impact of therapy on spermatogenesis depends on the type of treatment, but the time needed to recover testicular function following treatment with chemotherapeutic agents inducing azoospermia is unknown. STUDY DESIGN, SIZE, DURATION: In a retrospective study, the semen parameters of 519 patients (504 with sperm and 15 who were azoospermic) were investigated.HL patients were analysed before therapy. A longitudinal study was also conducted of semen quality in 202 patients pre- and post-ABVD (doxorubicin, bleomycin, vinblastine and dacarbazine) at T0 (baseline) and 6 (T6), 12 (T12) and 24 (T24) months after the end of treatment, and of 42 patients pre- and post-BEACOPP (bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, prednisone), COPP/ABVD (cyclophosphamide, vincristine, procarbazine, prednisone, doxorubicin, bleomycin, vinblastine and dacarbazine), OPP/ABVD (vincristine, procarbazine, prednisone, doxorubicin, bleomycin, vinblastine and dacarbazine) or MOPP (mechlorethamine, vincristine, procarbazine and prednisone) and inguinal radiotherapy at different observation times (from T0 to 16 years after treatment). PARTICIPANTS/MATERIALS, SETTING, METHODS: Semen parameters were examined according to World Health Organization 2010 criteria, evaluating sperm concentration, total sperm number, progressive motility and morphology. MAIN RESULTS AND THE ROLE OF CHANCE: Our data, which pertain to the largest caseload reported to date, indicate that 75% of HL patients are normozoospermic prior to treatment. The results from the HL patients studied pre- and post-therapy demonstrate that spermatogenesis recovery depends on the therapeutic regimen used. After ABVD, there was a statistically significant decrease in sperm concentration and total sperm number at T6 and T12 (P < 0.001; P < 0.01, respectively). There was a significant drop in progressive motility (P < 0.001) and a significant increase in abnormal forms (P < 0.01) at T6. The differences in sperm concentration, total sperm number and abnormal forms at T0 and T24 were not statistically significant, indicating that sperm quality had returned to pre-therapy values. The most interesting data in terms of patient management arise from the study of azoospermia induced by other chemotherapeutic agents. A high number of BEACOPP, COPP/ABVD, OPP/ABVD or MOPP cycles (≥6) induced a permanent absence of sperm in the seminal fluid, while even following a low number of cycles (<6), spermatogenesis only recovered after 3-5 years and semen quality was highly impaired. LIMITATIONS, REASONS FOR CAUTION: The study type (retrospective) and the low caseload and varying time of the follow-up do not permit any firm conclusions to be drawn about the recovery of spermatogenesis after BEACOPP or other combined therapies, or the identification of any risk factors for testicular function in treated patients. WIDER IMPLICATIONS OF THE FINDINGS: The pretreatment semen parameters of HL patients in this study were better than some results reported in the literature, with a higher percentage of normozoospermic patients. Strengths of this study were the large caseload of HL patients and a high degree of consistency in semen analysis, as all parameters were assessed in the same laboratory. Following the azoospermia induced by different chemotherapeutic protocols, spermatogenesis may take several years to recover. Awareness of this issue will enable oncologists to better inform patients about the possibility of recovering fertility post-treatment and also demonstrates the importance of semen cryobanking before beginning any cancer treatment. STUDY FUNDING/COMPETING INTERESTS: Supported by a grant from the Italian Ministry of Education and Research (MIUR-PRIN) and the University of Rome 'La Sapienza' Faculty of Medicine. The authors have no conflicts of interest.

Acute endothelial response to testosterone gel administration in men with severe hypogonadism and its relationship to androgen receptor polymorphism: a pilot study.

PURPOSE: Testosterone (T) exerts different effects on the cardiovascular system. Despite this knowledge, the acute vascular effect of androgen remains still poorly understood. METHODS: We investigated the acute effects of T on vascular function in ten men (18-40 years age) with hypogonadism and severe hypotestosteronemia [serum total testosterone (TT) = 0.6 ± 0.3 ng/mL]. In a 4-day double-blind, randomized, placebo-controlled crossover study, we administered 80 mg daily dose of transdermal-T gel (TG) and evaluated endothelial variations with Endopat2000 (reactive hyperemia index, RHI and the augmentation index, AI); also, CAG repeat polymorphism in exon 1 of the androgen receptor gene was investigated. RESULTS: After TG administration, RHI significantly improved at 4 h (p < 0.05), while AI improvement was recorded at 4 and 96 h, also when adjusted for heart rate (AI@75; p < 0.01 and p < 0.001, respectively). Direct relationships between ΔT, ΔDHT and ΔRHI variations (r = 0.37, p < 0.01; r = 0.17, p < 0.05, respectively) as well as between "CAG repeats" length and ΔLnRHI at 96 h (p < 0.03, r (2) = 0.47) were found. An inverse relationship between ΔT and ΔAI (p < 0.01, r = -0.35) and ΔAI@75 (p < 0.01, r = -0.38) were found. CONCLUSION: Administration of TG causes an acute vasodilation and improves arterial stiffness probably due to non-genomic actions of T. Endothelial vasodilatory response was more pronounced depending on higher plasma TT and DHT levels attained. Clinical implications in elderly frail populations are discussed.

Exposure to polychlorinated biphenyls and hexachlorobenzene, semen quality and testicular cancer risk.

PURPOSE: We carried out a case-control study to investigate the possible role of occupational and environmental exposure to endocrine disruptors in the onset of testicular cancer (TC). METHODS: We evaluated 125 TC patients and 103 controls. Seminal fluid examination and organochlorine analysis were performed in all subjects. Cases and controls were also interviewed using a structured questionnaire to collect demographic information, residence, andrological medical history and dietary information. RESULTS: We found that a higher level of reproductive tract birth defects was associated with a higher risk of TC. With regard to diet, cases reported a higher consumption of milk and dairy products than controls. Overall, there was a statistically significant increase in TC risk in cases with detectable values of total polychlorinated organic compounds against controls (14.4 vs. 1.0 %; p < 0.001). TC patients with detectable levels of organochlorines had lower mean semen parameters than those with undetectable levels, although this difference was not statistically significant. CONCLUSION: The International Agency for Research on Cancer recently included dioxin-like polychlorinated biphenyls (PCBs) in Group 1 of known human carcinogens. Our study confirmed and identified various risk factors for testicular cancer: cryptorchidism, consumption of milk and dairy products, parents' occupation and serum concentration of hexachlorobenzene and PCBs and, for the first time, we showed the correlation between semen quality and the serum concentration of these pollutants.

Hormone and genetic study in male to female transsexual patients.

BACKGROUND: Data of the literature demonstrated controversial results of a correlation between transsexualism and genetic mutations. AIM: To evaluate the hormone and gene profile of male-female (M-F) transsexual. SUBJECTS AND METHODS: Thirty M-F transsexuals aged 24-39. Seventeen had already undergone sex reassignment surgery, 13 were awaiting. All subjects had been undergoing estrogen and antiandrogen therapy. We studied hormones of the hypothalamus- pituitary-testicular axis, thyroid and adrenal profile, GH basal and after GHRH stimulation, IGF-I. The gene study analyzed SRY, AR, DAX1, SOX9, AZF region of the Y chromosome. RESULTS: Pre-surgery subjects had elevated PRL, reduced testosterone and gonadotropins. Post-surgery subjects showed reduced androgens, a marked increase in LH and FSH and normal PRL. Cortisol and ACTH were similar to reference values in pre- and post-surgery patients. There was a marked increase in the baseline and post-stimulation GH values in 6 of the 13 pre-surgery patients, peaking at T15. IGF-I was similar to reference values in both groups except for one post-surgery patient, whose level was below the normal range. There were no polymorphisms in the amplified gene region for SOX9, and a single nucleotide synonimous polymorphism for DAX1. No statistically significant differences were seen in the mean of CAG repeats between controls and transsexual subjects. SRY gene was present in all subjects. Qualitative analysis of the AZFa, AZFb, and AZFc regions did not reveal any microdeletions in any subject. CONCLUSIONS: This gender disorder does not seem to be associated with any molecular mutations of some of the main genes involved in sexual differentiation.

Are apolipoprotein E alleles correlated with semen quality?

Apolipoproteins have a unique role in lipoprotein metabolism regulation, aiding lipid transport and acting as a cofactor of the enzymes involved in metabolism. There are three co-dominant alleles, APOE*2, APOE*3 and APOE*4, which encode three protein isoforms, apoE2, apoE3 and apoE4. APOE*3 is the most frequent in all populations thus far investigated, ranging from 50 to 90%. Some studies have tried to resolve a genetic 'dilemma' by evaluating the cause of the frequency and survival of the three alleles. Genetic drift, migration or natural selection could explain the current distribution of APOE gene frequencies worldwide. If APOE*4 is the ancestral allele, APOE*3 must have offered a considerable selective advantage, perhaps consisting of a positive effect during the reproductive period. Given this, there is a need to understand if APOE gene polymorphism might affect reproductive capacity. Few studies have been conducted in this area, and they generally correlate APOE polymorphism with reproductive efficiency in terms of number of children. The aim of our study was to look for correlations between APOE polymorphism in humans and semen quality, to establish if APOE genotypes have any demonstrable effect on spermatogenesis. In conclusion, our data show that APOE polymorphism is not associated with semen quality, as it is present to a similar extent in both normal and impaired or absent spermatogenesis. This demonstrates once again that the use of number of children as an index of fertility is not indicative of real male reproductive capacity.

Protamine-1 and -2 polymorphisms and gene expression in male infertility: an Italian study.

BACKGROUND: Correct histone/protamine replacement is an important stage in chromatin condensation during spermiogenesis in humans. There are two types of protamines: protamine 1 (P1) and the protamine 2 family (P2, P3, and P4), coded by the genes PRM1 and PRM2. AIM: We analyze the sequences and gene expression of PRM1 and PRM2 and their relationship with defective spermatogenesis. MATERIALS AND METHODS: Sequence analysis was carried out on 163 patients attending our laboratory for analysis of seminal fluid. Patients were divided into three groups: normozoospermic (53), teratozoospermic (60), and azoospermic (50). Gene expression was analyzed in seven patients with azoospermia and one with cryptozoospermia. RESULTS: Seven single nuclotide polymorphisms (SNP) were identified: G54A, G102T and C230A for PRM1, and C246T, G288C, G298C and C373A for PRM2. For C230A, the CA genotype was present in 38% of teratozoospermic vs 55% of normozoospermic and 64% of azoospermic patients; for C373A, CA was found in 37% of teratozoospermic vs 47% of normozoospermic and 64% of azoospermic patients. In contrast, for G298C, GC was more common in the teratozoospermic (63%) than in the normozoospermic (49%) or azoospermic (48%) groups. These differences could suggest a greater susceptibility of these patients to abnormal sperm morphology. In five patients the levels of transcripts were reduced with respect to the control. CONCLUSION: These data suggest that premeiotic arrest is associated with extremely reduced protamine expression. New studies of both PRM1 and PRM2 and their mRNA expression could help us better understand the molecular mechanisms underlying the protamine transcription and translation processes.

Effects of a dietary supplement on chronic pelvic pain syndrome (Category IIIA), leucocytospermia and semen parameters.

Chronic prostatitis (CP) is one of the most common male urogenital diseases and a significant public health problem in industrialised countries. It is associated with a low quality of life and significant expense. Given the poor results achieved with antibiotics, scientific interest has turned to the use of natural substances with a known activity on prostate function. The aim of our study was to evaluate the effect of a new dietary supplement containing lycopene, epigallocatechin gallate, ellagic acid, selenium and zinc on semen parameters and on leucocyte concentration in seminal fluid and expressed prostate secretion (EPS) in patients with CP without infection [National Institute of Health (NIH) Category IIIA], in comparison with a control group with the same condition who did not undergo any treatment during the study period. Our data showed a statistically significant reduction in inflammatory parameters (leucocytes in seminal fluid and EPS) and a statistically significant improvement in progressive sperm motility and sperm morphology in patients treated with the supplement in comparison with the untreated group. Improvements were also seen in the pain score of the NIH-Chronic Prostatitis Symptom Index (CPSI), confirming that the reduced inflammation also resulted in a reduction in pain.

Semen quality in men with Y chromosome aberrations.

Infertile males sometimes bear structurally balanced chromosome aberrations, such as translocations and inversions, which involve both autosomes and sex chromosomes. The aim of this study was to evaluate genotype-phenotype correlations in a sample of infertile men with various types of Y chromosome abnormalities. In particular, we examined the effect of (i) balanced structural aberrations such as translocations between sex chromosomes and autosomes; (ii) unbalanced structural aberrations such as deletions or isodicentrics, both [idic(Yp)] and [idic(Yq)]. We studied 13 subjects bearing Y chromosome aberrations. Each patient underwent seminal fluid examination, andrological inspection, hormone study, testicular ultrasound, conventional and molecular cytogenetic analysis and study of Y chromosome microdeletions. Comparison of genotype and sperm phenotype in infertile patients with various Y chromosome aberrations revealed the key role of meiotic pairing defects in arresting spermatogenesis, both in the presence and in the absence of azoospermic factor microdeletions and cell mosaicism. The failure of meiosis and, in consequence, spermatogenesis may be a result of the failure to inactivate the X chromosome in the meiotic prophase, which is necessary for normal male spermatogenesis to take place.

Sumo1-ylation of human spermatozoa and its relationship with semen quality.

Sumoylation is a post-translational modification involved in the regulation of several cell functions. Recent studies suggest its involvement in spermatogenesis, but occurrence and function of SUMO (small ubiquitin-like modifier) in mature spermatozoa remain unknown. We report the occurrence of several SUMO1-conjugated proteins, in a range of 20-85 kDa, in ejaculated spermatozoa. By cytofluorimetric analysis, we evaluated the percentage of SUMO1-positive spermatozoa in 58 subjects undergoing semen analysis in our laboratory and correlated the obtained values with semen parameters. We found that the percentage of SUMO1-positive spermatozoa was inversely correlated with total (r = -0.35, p < 0.01) and progressive motility (r = -0.29, p < 0.05). Such correlations become stricter when only asthenospermic subjects were included in the analysis (r = -0.58, p = 0.01 for progressive motility, n = 17) and were lost in non-asthenospermic subjects. By immunofluorescence and immunoconfocal fluorescence, we demonstrated that SUMO1 is mainly located in the nucleus and, occasionally, in the midpiece of spermatozoa. Immunoelectron microscopy as well as a long permeabilization protocol demonstrated a massive localization of SUMO-1 in the nucleus. By using a fluorescent probe to distinguish dead/live cells, we show that SUMO1 is mainly present in live spermatozoa. In conclusion, sumoylation of human spermatozoa may be involved in the regulation of motility.

Strategies and advantages of early diagnosis in Klinefelter's syndrome.

Nearly 70 years after its description, Klinefelter's syndrome (KS) remains a largely undiagnosed condition. In addition to its typical characteristics of increased follicle-stimulating hormone secretion and small and firm testes, the syndrome presents an extremely wide spectrum of phenotypes. This could be explained by the possible presence of chromosomal mosaicism, androgen receptor polymorphisms and related heterogeneous endocrine abnormalities. The varied but relatively mild physical abnormalities also explain why many patients do not receive clinical attention until adulthood, when they seek medical advice on small testes or infertility. Diagnosis is also hindered by the low awareness of the disease among health professionals. This paper aims to review the possible signs of KS at different stages of life that could help achieve an early (or at least earlier) diagnosis. It has been demonstrated that the early diagnosis of KS improves patients' quality of life and enables better medical treatment. To achieve this, it is crucial to increase both medical and general awareness of the disease, including through use of the media and patients' associations.

Efficacy of recombinant human follicle stimulating hormone at low doses in inducing spermatogenesis and fertility in hypogonadotropic hypogonadism.

BACKGROUND: Recombinant-FSH (rFSH) added to hCG at dose of 450 IU weekly is effective in inducing spermatogenesis in patients with hypogonadotropic hypogonadism (HH), but there are no data on the use of lower doses. AIM: This observational retrospective study evaluated whether 150-225 IU of rFSH weekly were able to induce spermatogenesis in HH men who failed to start it with hCG alone. SUBJECTS AND METHODS: Thirty-four patients with pre-pubertal onset HH (20-44 yr old) without adverse fertility factors were considered for this study. After hCG pre-treatment they received also either rFSH (Group 1) or highly purified urinary FSH (hpFSH) (Group 2) 75 IU sc 2 or 3 times weekly. Semen analysis was performed every 3 months during pre-treatment and the 1st yr of combined therapy. Patients were also invited to refer pregnancies in their partners during the subsequent 12 months. RESULTS: Total sperm count/ejaculate did not show significant difference between 2 groups, while a significantly higher forward motility was observed in Group 1 (p<0.05). The median times to achieve sperm output thresholds (first sperm appearance, sperm concentration >1.5 or >5 mil/ml) were significantly lower in Group 1 (p<0.04, 0.03, and 0.001, respectively). A tendency to a shorter time to pregnancy was shown in partners of Group 1. CONCLUSIONS: Our data indicate that lower rFSH week dose than that so far used was able to induce potentially fertilizing sperm output in HH men previously treated with hCG. The rFSH effects are comparable to those of hpFSH but with a trend to a faster outcome achievement.

Expression of Fas and Fas ligand in human testicular germ cell tumours.

In the present study, we analysed the expression of Fas ligand (FasL) and its cognate receptor Fas in 14 seminomatous testicular germ cell tumours (TGCT) and six normal testicular tissues obtained following orchiectomy. Tissue samples have been processed to prepare either total RNA or protein extracts or fixed and embedded in paraffin for immunohistochemistry (IHC) experiments. Quantitative RT-PCR experiments demonstrated in TGCT a significant (p < 0.01) increase of the FasL mRNA expression of 21.1 +/- 5.4 fold, with respect to normal tissues. On the contrary, in the same cancer tissues, the levels of Fas mRNA were significantly (p < 0.01) reduced to 0.27 +/- 0.06 fold. These observations were confirmed in western blot experiments showing a significant increase of FasL and a concomitant decrease of Fas proteins in testicular cancer tissues, with respect to normal testis. Moreover, IHC experiments showed a strong FasL immuno-reactivity in six out of eight TGCT samples analysed, while Fas immuno-positivity was found in cancer cells of only two TGCT tissues. In addition, in all tumour samples, infiltrating lymphocytes were Fas positive. However, no correlation could be observed between Fas or FasL mRNA variations and clinical parameters such as patient's age, TNM stage or tumour size. We also compared the serum levels of soluble FasL (sFasL) of 15 patients affected by seminomatous TGCT, of four patients with non-seminomatous TGCT and six age-matched healthy males. No significant differences in sFasL serum level could be identified. In conclusion, our data demonstrated that the majority of seminomas are characterized by an increased expression of FasL and a concomitant reduction of Fas, with respect to human normal testis, and that sFasL serum level is not a tumour marker for patients affected by TGCT.

Epidemiology, diagnosis, and treatment of male hypogonadotropic hypogonadism.

Hypogonadotropic hypogonadism (HH), or secondary hypogonadism, is a clinical condition due to an impairment of the pituitary function, characterized by low testosterone plasma levels associated with normal or low FSH and LH plasma levels. An impairment of gonadotropin secretion and, therefore, a reduced efficiency of spermatogenesis was reported to be frequently associated to conditions different from the classical causes of secondary hypogonadism. These conditions (metabolic, endocrine and eating disorders, physical exercise etc.) have been associated with a non-classical form of HH that could be called "functional" HH (FHH). FHH differs from the classical one by the evidence that gonadotropin levels are in the low-normal range, but are inadequate for the testosterone levels, that often are also in the low-normal range. This commentary aims at reviewing knowledge on the forms of male HH in order to indicate and discuss clinical context, diagnostic and therapeutic approach in the less known non-classical form, i.e. FHH.

Testicular function in boys previously treated with recombinant-human growth hormone for non-growth hormone-deficient short stature.

Data on the effects of recombinant human GH (hGH) therapy during male puberty on future testis function are still inconclusive. The aim of this study was to investigate the long-term effects of recombinant hGH treatment on reproductive function in non-GH-deficient short stature boys. Eight boys with non-GH-deficient short stature, affected by constitutional delay of puberty or idiopathic short stature, were retrospectively studied after recombinant-hGH treatment to verify gonadal development, hormone production and semen quality. Auxological data, endocrinological/ andrological parameters and laboratory evaluation (GH, IGF-I, FSH, LH, testosterone, inhibin B) were assessed before treatment; after completion of pubertal development, the same parameters plus SHBG levels were evaluated and a seminal fluid examination was conducted (ejaculate volume, pH, sperm concentration, total sperm count, forward and total motility, morphology). All patients showed normal testicular volume at the final pubertal stage, with regular androgenization. Hormonal levels were within the normal adult range in all boys. Considering the immature reproductive system of these patients in comparison with adults, semen parameters (sperm count, motility, and morphology) were within almost normal limits, except in one patient. Although patients showed the wide fluctuation of semen values frequently observed at the end of puberty, the hypophysis-gonadal axis hormones were in the normal range in all adolescents. Pathological measurements of some seminal parameters were found in one patient only. This study suggests that recombinant hGH treatment has no detrimental effects on the development and maturation of male gonadal function in non- GH deficient short stature young patients.

Testicular cancer and sperm DNA damage: short- and long-term effects of antineoplastic treatment.

The aim of this study was to investigate sperm DNA damage induced by chemo- and radiotherapy in patients with testicular cancer to provide data on the extent and persistence of nuclear damage that might affect individual reproductive potential. We evaluated pre- and post-antineoplastic treatment sperm DNA integrity, expressed as DNA Fragmentation Index (DFI), in a large caseload of testicular cancer patients by sperm chromatin structure assay. The mean total DFI for all patients at T0 was 18.0 ± 12.5%. Sperm chromatin profile was markedly impaired at T3 (27.7 ± 17.4%) and T6 (23.2 ± 15.3%), improving considerably at T12 and T24 (14.0 ± 8.9% and 14.4 ± 10.3%). After chemotherapy, we found a marked increase in DFI at T3 and T6 and a significant reduction at T12 and T24 in comparison with the baseline. In contrast, DFI increased at T3 and T6 after radiotherapy but the subsequent reduction was far less marked, reaching baseline values at T12 and T24. Finally, post-treatment DNA damage was not age or histotype dependent, but was more marked in the advanced stage of cancer. In this study, we showed that the chromatin profile may be affected in the months immediately following the end of the treatment, improving after 12-24 months. Our results thus indicate that post-treatment DNA damage is influenced both by the type and intensity of the therapy and by the pathological and clinical stage of the disease.

Comparison between primary sex ratio in spermatozoa of bulls and secondary sex ratio in the deriving offspring.

The objectives of the present work were to compare the primary sex ratio in sperm with the secondary sex ratio recorded in the offspring produced by artificial insemination (AI) with the same sperm and assess whether the primary sex ratio is influenced by sperm survival and motility after thawing. Calving data of 98 Holstein Friesian bulls used in AI were collected during 4 years, and commercial semen of the same bulls was analyzed immediately after thawing and after swim-up using a real-time polymerase chain reaction method developed and validated in our laboratory. Calving data relative to single bulls did not reveal any significant deviation between genders from the theoretical 1:1 for none of the bulls, being the mean values of male and female calves born 52.1 ± 2.80% and 47.9 ± 2.71%, respectively. Thereafter, calving events of bulls were classified and analyzed according to four classes of years: 2009 (n = 13,261), 2010 (n = 21,551), 2011 (n = 24,218), and 2012 (n = 41,726), and seasons categorized as winter, spring, summer, and fall. When data aggregated per years were analyzed, the difference between the two sexes was significant (P < 0.005) in favor of the male gender, whereas no influence of the season was evidenced. Real-time polymerase chain reaction did not evidence any difference between the mean values of frequency of Y chromosome-bearing sperm detected in three sperm batches of the same bulls analyzed immediately after thawing (51.1 ± 2.1), nor a difference with respect to the theoretical 1:1 ratio was reported after sperm analysis of one batch of sperm of the bulls analyzed after swim-up and immediately after thawing (50.1 ± 2.1 and 49.8 ± 1.8, respectively). The results are consistent with the observation of the farmers who often report a skewed sex ratio of the calves being born with AI in favor of the male gender. However, we have not evidenced differences in the primary sex ratio with respect to the theoretical 1:1 ratio both at thawing and after swim-up, thus demonstrating that the freezing procedure itself does not impact selectively on the survival of the X or Y chromosome-bearing sperm. Therefore, we hypothesize that the difference between genders observed after AI is more likely due to the events occurring after fertilization, which can comprise an impaired function of the X- or Y-bearing sperm with consequences on embryo development or a maternal influence.

Androgen receptor polymorphisms and testicular cancer risk.

Testicular cancer (TC) is currently the most common malignant solid tumour in Caucasian males aged 15-39 years. Epidemiological evidence suggests that its onset may be due to an imbalance in the action of steroidal sex hormones and their receptors. A faulty androgen receptor signalling pathway can, in fact, cause various male reproductive disorders. The androgen receptor (AR) gene has two polymorphic segments consisting of CAG and GGC repeats. The length of CAG repeats has been shown to affect the regulation of AR activity. In our study, we used fragment analysis to evaluate the AR gene repeats of 302 TC patients and 322 controls, to establish if there is any association between repeat number and TC. This study of the largest Italian caseload investigated to date highlighted three particularly significant aspects. First, a CAG repeat number of ≥25 may be considered a risk factor for the onset of TC, given its greater frequency in patients in comparison with controls. This difference became significant for the non-seminoma group. Second, men with CAG repeats below 21 or above 24 were found to have a, respectively, 50 and 76% higher risk of TC than those with CAG 21-24, suggesting that these too can be considered a risk factor for TC. Finally, stage II patients were more likely to have a CAG repeat number <21 or >24 than stage I patients.

Lipoperoxidation damage of spermatozoa polyunsaturated fatty acids (PUFA): scavenger mechanisms and possible scavenger therapies.

The lipid metabolism in sperm cells is important both as one of the main sources for energy production and for cell structure. The double leaflets of the membrane should be considered not simply as a passive lipid film, but as a very specialized structure. The complete maturation of the sperm cell membrane is attained after testicular lipid biosynthetic processes and after passage through the epididymis. A special composition of membrane phospholipids, rich in polyunsaturated fatty acids (PUFA), and the different composition of sperm and immature germ cell membrane are described and discussed. Testis germ cells as well as epididymal maturing spermatozoa are endowed with enzymatic and non-enzymatic scavenger systems to prevent lipoperoxidative damage. Catalase, superoxide dismutase and GSH-dependent oxidoreductases are present in variable amounts in the different developmental stages. Phospholipid hydroperoxide GSH peroxidase (PHGPx) activity and alpha tochopherol of epididymal spermatozoa are considered in detail. Their distribution and roles in caput and cauda epididymal sperm cells are discussed. Seminal plasma also has a highly specialized scavenger system that defends the sperm membrane against lipoperoxidation and the degree of PUFA insaturation acts to achieve the same goal. Systemic predisposition and a number of pathologies can lead to an anti-oxidant/pro-oxidant disequilibrium. Scavengers, such as GSH, can be used to treat these cases as they can restore the physiological constitution of PUFA in the cell membrane. The results of GSH therapy are presented and discussed.

Sperm parameters after early left varicocele treatment.

OBJECTIVE: We evaluated whether early treatment for left varicocele can improve sperm parameters. DESIGN: Patient follow-up and comparison with control groups. SETTING: Surgical department of a Children's Hospital and a university infertility-care center. PATIENT(S): Nineteen patients who underwent surgery for left varicocele as adolescents (mean +/- SD, 14.11 +/- 1.59 years) were followed up as adults (mean +/- SD, 19.37 +/- 1.16 years). MAIN OUTCOME MEASURE(S): A testicular examination was performed and varicocele staging was done at the time of surgery. Andrologic examination and semen analysis were done 2-8 years later. Semen results were compared with those of two age-matched control groups of 19 healthy, randomly selected subjects and 19 unoperated patients suffering from left varicocele of the same degree. RESULT(S): The mean values of sperm parameters in the patient study group were similar to those of the healthy controls except that the sperm concentration was significantly lower. However, sperm motility and morphology mean values in the patient study group were significantly higher than those of the unoperated patients. CONCLUSION(S): This study indicates that surgery in adolescence can be useful, possibly because the testis is developing at that age. Large-scale, double-blind controlled trials on early treatment of left varicocele are needed to identify possible markers of disease severity.

Follicular fluid and human granulosa cell cultures: influence on sperm kinetic parameters, hyperactivation, and acrosome reaction.

OBJECTIVE: To evaluate the influence of human granulosa cell (GC) cultures and follicular fluid (FF) on sperm kinetic parameters, hyperactivation, and the acrosome reaction compared with the influence of human tubal fluid (HTF) and Ham's F-10 medium. DESIGN: Sperm kinetic parameters, hyperactivation, and the acrosome reaction were evaluated after 6 hours of incubation in HTF, Ham's F-10 medium, FF, and GC cultures. SETTING: Infertility and In Vitro Fertilization Centre, Reproductive Endocrinology Unit, Institute of Obstetrics and Gynaecology. PATIENT(S): Sixteen normal semen samples. INTERVENTION(S): Sperm kinetic parameters and hyperactivation were analyzed using an automated videomicrography system, the acrosome reaction was performed using a triple-stain technique, and progesterone and 17OH-progesterone levels were measured with the use of commercially available kits. MAIN OUTCOME MEASURE(S): Sperm kinetic parameters, hyperactivation, acrosome reaction. RESULT(S): The percentage of motile sperm, the mean curvilinear velocity, and the mean of the maximum amplitude of lateral head movement were increased significantly after 6 hours of incubation in FF or GC cultures compared with incubation in HTF or Ham's F-10 medium, whereas the mean linearity was decreased significantly. Follicular fluid and GC cultures significantly increased hyperactivation and the acrosome reaction compared with the values obtained using HTF and Ham's F-10 medium. Progesterone and 17OH-progesterone levels were increased significantly after incubation in FF and GC cultures compared with HTF and Ham's F-10 medium. CONCLUSION(S): Follicular fluid and GC cultures increase sperm motility parameters, hyperactivation, and the acrosome reaction. This effect may be related to GC detoxification of the microenvironement or GC secretion of peptides, glycoproteins, growth factors (insulin-like growth factors 1 and 2), or steroids (progesterone and 17OH-progesterone).