Domenico Milardi

Male fertility and reduction in semen parameters: A single tertiary-care center experience

Background. Infertility is both a clinical and a public problem, affecting the life of the couple, the healthcare services, and social environment. Standard semen analysis is the surrogate measure of male fertility in clinical practice. Objective. To provide information about the relationship between semen parameters and spontaneous conception. Methods. We evaluated retrospectively 453 pregnancies that occurred among 2935 infertile couples evaluated at an infertility clinic of a tertiary-care university hospital, between 2004 and 2009. Results. Normal semen analysis was present only in 158 patients; 295 subfertile patients showed alterations in at least one seminal parameter. A reduction in all seminal parameters was observed in 41 patients. Etiological causes of male infertility were identified in 314 patients. Conclusion. Our data highlights the possibility of a spontaneous conception with semen parameters below WHO reference values. Therefore, we support the importance of defining reference values on a population of fertile men. Finally, we analyzed the related ethical issues.

Proteomic approach in the identification of fertility pattern in seminal plasma of fertile men

OBJECTIVE To identify a panel of common seminal proteins in human seminal plasma by fertile men that might be involved in successful reproduction. DESIGN Experimental study. SETTING University hospital. PATIENT(S) Five fertile men who conceived within 3 months before the start of the study. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Proteomic analysis performed by an Ultimate 3000 Nano/Micro-HPLC apparatus equipped with an FLM-3000-Flow manager module and coupled with an LTQ Orbitrap XL hybrid mass spectrometer; gene ontology analysis. RESULT(S) From 919 to 1,487 unique proteins were identified per individual subject sample. Among these proteins, 83 proteins were present in all samples, including some proteins that might be involved in male fertility, such as semenogelin I, semenogelin II, olfactory receptor 5R1, lactoferrin, hCAP18, spindling, and clusterin. The gene ontology annotation analysis provided further information in describing common pattern in male fertility. CONCLUSION(S) The identification of common seminal plasma proteome in fertile men could provide better insight into the physiology of male fertility and might identify novel markers of male infertility.

Relationship between sperm cell ubiquinone and seminal parameters in subjects with and without varicocele

Summary. In a previous paper it was demonstrated that Coenzyme Q10, a lipidic molecule with important antioxidant properties, is present at remarkable levels in human seminal fluid, and shows a direct correlation with seminal parameters (sperm count and motility). In patients with varicocele, on the contrary, correlation with sperm motility was lacking and a higher proportion of Coenzyme Q10 was found in seminal plasma. In the present study, the levels of Coenzyme Q10 in the cell pellet of spermatozoa, obtained after centrifugation of semen, were evaluated. In nonvaricocele subjects it was observed that a higher concentration of Coenzyme Q 10 (expressed as ng of the molecule per million of cells) was present in the spermatozoa of oligospermic and asthenospermic patients (sperm count <20*106 spermatozoa ml−1, sperm motility <40%). This relationship was not observed in varicocele subjects, who also showed slightly lower intracellular absolute values of the coenzyme.

Proteomics of human seminal plasma: Identification of biomarker candidates for fertility and infertility and the evolution of technology

Proteomics is a research area that has developed rapidly in the last decade. It studies the large‐scale characterization of the full protein components of a cell, a tissue, or a biological fluid. In the last decade, clinical proteomics has developed new technology and bioinformatics useful in identifying molecular markers of pathology; the next decade might be the era of proteomics. Seminal plasma (SP) represents a good sample for proteomic analysis in the evaluation of male fertility/infertility. SP is an acellular fluid conglomerate, comprised of contributions from the epididymis and accessory sexual glands. Human SP contains many proteins that are important to the successful fertilization of the oocyte by the spermatozoa. Proteomic studies have identified numerous seminal‐specific proteins, and recent reports have provided a further understanding of their function with respect to male fertility. Upon further validation, these proteins may be useful in the clinical distinction between fertility and infertility. This article reviews the proteomic methods, such as one dimensional polyacrylamide gel electrophoresis (1D–PAGE), two‐dimensional polyacrylamide gel electrophoresis (2D–PAGE), and mass spectrometry (MS), employed to detect human SP markers involved in fertility and infertility. As such, proteomic studies will help the development of new techniques to identify novel biomarkers for a better clinical diagnosis and treatment of male infertility. Mol. Reprod. Dev. 80: 350–357, 2013.

Effects of ghrelin administration on endocrine and metabolic parameters in obese women with polycystic ovary syndrome

Introduction: The novel peptide ghrelin displays multiple endocrine and non-endocrine actions. Its strong GH-releasing activity in humans has long been recognized. However, in obesity, ghrelin administration induces a blunted GH secretion, enhances glucose and reduces insulin levels. The effects of ghrelin administration have not been investigated in polycystic ovary syndrome (PCOS), which can be associated with obesity, hyperinsulinism, and GH hyposecretion. Leptin is a mediator for energy balance opposed to ghrelin; both of them are supposed to act as regulators of reproductive functions. Aim of the study: Evaluate the endocrine and metabolic response to ghrelin administration in PCOS obese patients compared to body mass index (BMI)-matched and normal weight women. Materials and methods: Nine obese PCOS patients (BMI: 35.4±1.2 kg/m2) (OB PCOS), 6 obese controls (BMI: 38.4±1.1 kg/m2) (Ob), and 6 normal-weight women (BMI: 23±0.6 kg/m2) (NW) were enrolled in the study. In all patients we performed: 1) basal hormonal evaluation including FSH, LH, estradiol, testosterone, androstenedione, DHEAS, SHBG, 17-hydroxyprogesterone (17OHP), IGF-I, free T3 (FT3), free T4 (FT4) and ghrelin levels; 2) metabolic evaluation as follows: concentration of non-esterified fatty acid (NEFA) and oral glucose tolerance test (OGTT) (75 g); homeostasis model assessment (HOMA); glucose and insulin response to ghrelin administration (1 µg/kg); 3) measurement of GH, PRL, TSH, and leptin levels after infusion of ghrelin. Results: Administration of ghrelin increased glucose and reduced insulin levels in both Ob and OB PCOS. Moreover, ghrelin enhanced GH and PRL levels in all groups but it did not modify TSH and leptin levels. GH peak and area under the curve (AUC) in OB PCOS and Ob were lower than controls (p<0.05). Similar PRL peak and AUC values were observed in all groups. Conclusions: In both obese and PCOS obese patients, leptin levels are not influenced by ghrelin administration. Moreover, the GH response after ghrelin administration is blunted. However, ghrelin exerts glucose-enhancing and insulin-lowering effects, the latter absent in NW.

The effect of treatment with growth hormone on fertility outcome in eugonadal women with growth hormone deficiency: report of four cases and review of the literature.

OBJECTIVE To highlight the clinical role of standard GH replacement treatment on fertility and pregnancy outcomes in four infertile eugonadal women with GH deficiency (GHD). DESIGN Case report. SETTING Department of endocrinology and infertility clinic, tertiary-care university hospital. PATIENT(S) Four normogonadotrophic, normoprolactinemic patients with long-standing infertility, affected by GHD. In two patients (aged 30 and 34 years) GHD was diagnosed after a brain injury. The third patient (age 30 years) had a primary empty sella, documented by magnetic resonance imaging of the pituitary. The last patient (age 28 years) underwent transsphenoidal surgery for Ratkes cyst. The LH and FSH responses to GnRH were normal in all four patients. Two of the four patients also had secondary hypoadrenalism and hypothyroidism. INTERVENTION(S) Patients received recombinant human GH replacement therapy (0.9-1.8 mg/week) for 6-12 months until pregnancy was first indicated by biochemical markers (beta-hCG) and later confirmed by transvaginal sonography. The GH therapy was discontinued after confirmation of pregnancy. MAIN OUTCOME MEASURE(S) Pregnancy. RESULT(S) All patients remained off treatment throughout pregnancy; they had uneventful pregnancies and term deliveries. The babies were healthy and normal in terms of length and weight. CONCLUSION(S) Our case studies confirm the important clinical role of the GH-insulin-like growth factor I system in oocyte fertilization and the beginning of pregnancy in a selected population of eugonadotrophic infertile women.

Increased estradiol levels in venous occlusive disorder: a possible functional mechanism of venous leakage.

Venous insufficiency of the corpora cavernosa is the second most common cause of erectile dysfunction (ED). A functional insufficiency of the venous system has been hypothesised, but the cause is still unclear. To evaluate a possible endocrine mechanism, we have studied hormone profile in a group of nine patients with pure venous-leakage (VL) compared with a control group of 15 patients with ED of different origin. Prolactin, testosterone and gonadotropin levels did not differ among the two groups, while estradiol (E2) plasma concentration was significantly higher in VL patients compared to controls. Our data support the hypothesis that the steroid environment, in particular estradiol level, can influence venous vascular tone (via VEGF or NO), thus affecting venous leakage dysfunction. This point can explain a possible link between the high estradiol levels and a functional insufficiency of the venous system in ED.

The treatment of neuroendocrine tumors with long-acting somatostatin analogs: A single center experience with lanreotide autogel

The aim of this retrospective study was to evaluate the efficacy, safety, and tolerability of lanreotide autogel given to metastatic well-differentiated (WD) neuroendocrine tumors (NET) patients observed in our Institute between 2005 and 2008. Patients with metastatic NET referred to our tertiary referral center were given lanreotide autogel 120 mg/month by deep sc injection for a period of at least 24 months. The efficacy was evaluated by the relief of disease symptoms, behavior of tumor markers and response rate in terms of time to tumor progression. Safety and tolerability were evaluated by assessing the onset of adverse events and treatment feasibility. Twenty-three patients (13 males), median age 62 yr (range 32–87) were considered for the study. All patients were affected by WD metastatic NET and had tumor progression in the last 6 months before the enrolment in the study. Median duration of response was 28 months (range 6–50 months). Fourteen patients (60.9%) showed flushing and diarrhea which improved by 85.7% and 55.6%, respectively, bronchoconstrinction and abdominal pain also ameliorated. A complete, partial or no-changed response in the tumor markers behavior was observed, respectively, in 42.9%, 22.9%, and 17.1% of cases. According to RECIST (Response Evaluation Criteria In Solid Tumors) criteria (version 1.1), there were 2 partial regression (8.7%) and 15 stable disease (65.3%); 6 patients (26.0%) progressed. No patient complained from any severe adverse reaction. The results of our study suggest that lanreotide autogel is effective in the symptoms, biochemical markers, and tumor progression control of WD metastatic NET and confirm that the treatment is well tolerated.

Increased Total Antioxidant Capacity in Seminal Plasma of Varicocele Patients: A Multivariate Analysis

To investigate seminal antioxidant systems and their correlation with hormonal pattern in varicocele patients, we studied 33 varicocele (VAR) patients (12 oligozoospermic, 21 normozoospermic) and 34 non-VARs (10 idiopathic oligozoospermic, 24 normozoospermic). Non-enzymatic total antioxidant capacity (TAC) was measured using H2O2-metamyoglobin, which generates the radical form, spectroscopically detectable, of the chromogen 2,2′,-azinobis-3-ethylbenzothiazoline-6-sulphonate (ABTS): time till appearance (Lag) of ABTS·− signifies antioxidant concentration. Lag was significantly longer in VARs than controls suggesting ineffective utilization of antioxidants. A significant direct correlation (r = 0.65, p < 0.01) of Lag with sperm count was observed in non-VARs, while in VARs it was inverted, as well as with hematic FSH levels. A multivariate analysis including FSH, Lag, progressive spermatozoa, oligozoospermia and varicocele indicated a strong inverse correlation between FSH and motility (r2 = 0.31, p > F = 0.0007), not modified by Lag (r2 = 0.31, p > F = 0.002). Their inverse correlation with Lag may suggest that higher FSH levels, improving sperm antioxidant efficiency, counterbalance varicocele-induced dyspermia.

Soluble urokinase-type plasminogen activator receptor as a putative marker of male accessory gland inflammation.

The association between male accessory gland infection/inflammation (MAGI) and infertility is well‐known in clinical practice. Standard semen analysis, leukocytospermia, and microbiological tests are often not enough accurate for a diagnosis. A large amount of biochemical parameters in seminal plasma have been suggested as inflammation markers, however, there is not yet a sensitive and specific biomarker that accurately identifies MAGI. We investigated the presence of soluble urokinase‐type plasminogen activator receptor (suPAR), known marker of systemic inflammation, in the seminal plasma to evaluate its possible involvement in urogenital tract inflammation. On the basis of andrological evaluation, including spermiogram and ultrasound findings, we selected 76 patients with MAGI and 30 healthy men as control group. Patients were classified according to the results of the semen culture in group A (n = 28) presenting a bacterial MAGI and group B (n = 48) with abacterial MAGI. C‐reactive protein (CRP), total protein (TP), procalcitonin (PCT), leukocytes peroxidase (LP), and suPAR concentrations were assayed on seminal plasma. Spermiogram parameters were significantly lower in the patients with MAGI than in controls. CRP, TP, PCT, and LP did not differ in MAGI vs. controls. suPAR was detectable in all semen samples; it was significantly increased in A and B groups (86.6 ± 30.7 ng/mL vs. 39.7 ± 17.2 ng/mL) with an inverse correlation with sperm parameters. We selected by receiver operating characteristic curve a suPAR cut‐off value of 55.3 ng/mL as a diagnostic threshold for the diagnosis of MAGI. We report in this study the first evidence of suPAR presence in seminal plasma, focusing on its interesting role as reliable and sensitive marker of inflammation for the differential diagnosis of MAGI.