Arcangelo Barbonetti

Low birth weight and later development of insulin resistance and biochemical/clinical features of polycystic ovary syndrome

Reduced insulin sensitivity in adult life has been reported in subjects born at term small for gestational age (SGA) and in those born prematurely with very low birth weight (LBW) (<1,500 g). We assessed whether LBW (<2,500 g) young women, irrespective of whether they were born SGA or adequate for gestational age (premature AGA), exhibited a reduction in insulin sensitivity through a prospective historical design. The risk of developing biochemical and clinical features of polycystic ovary syndrome was also investigated. The study population included 35 LBW women (19 SGA [BW range, 1,000-2,400 g] and 16 premature AGA [BW range, 1,700-2,440 g]) aged 21.8 +/- 1.8 years and 35 term AGA controls, of similar age, recruited from a neonatal registry. All women underwent clinical, ultrasonographic, hormonal, and metabolic evaluations, including the composite insulin sensitivity index. Women under hormonal contraception (21.4%) were excluded from hormonal and metabolic analyses. Composite insulin sensitivity index was significantly lower in LBW women even when the 2 LBW subgroups, SGA and premature AGA, were analyzed separately (4.4 +/- 2.2 and 4.0 +/- 1.7, respectively) than in controls (6.9 +/- 4.4). The LBW women showed a significantly higher incidence proportion of irregular menses (14/35 [40%] vs 2/35 [5.7%]) and a significantly higher free androgen index (5.8 +/- 3.5 vs 3.9 +/- 3.2). They also showed a nonsignificantly higher proportion of hirsutism, acne, and polycystic ovaries. In conclusion, LBW (<2,500 g) young women, irrespective of whether they were SGA and premature AGA, exhibited a reduction in insulin sensitivity as compared with born at term AGA women. Furthermore, they exhibited an increased risk of developing clinical and biochemical features of polycystic ovary syndrome.

Erectile Dysfunction is the Main Determinant of Psychological Distress in Men with Spinal Cord Injury

INTRODUCTION The weight of erectile dysfunction (ED) among the various determinants of psychological distress in men with spinal cord injury (SCI) remains to be clarified. AIM The aim of this article was to evaluate psychological distress features in SCI men with or without ED. METHODS Forty consecutive patients with neurologically stable SCI were included in the study. Functional independence (FI) was assessed by Barthel Index (BI), which was divided into global score (questions 1-10) and bowel/bladder subscore (questions 5 and 6). Erectile function was evaluated with Sexual Health Inventory for Men (SHIM). MAIN OUTCOME MEASURES Psychological distress was assessed with the Symptom Checklist-90-revised (SCL-90-R), scoring nine primary dimensions and their combination as Global Severity Index, a global index of psychological distress. RESULTS All SCL-90-R scores and the percentage of patients with scores >75th percentile of the entire study population were significantly higher in the group with ED (N=21) than without ED (N=19). Most of SCL-90-R subscales were inversely correlated with SHIM score. ED was exhibited by a high proportion (84%) of men with thoracolumbar lesions but by no patients with cervical lesions. Men with cervical lesions exhibited significantly lower SCL-90-R scores than those with thoracolumbar lesions, in spite of lower FI. However, the thoracolumbar group also reported a more severe bowel/bladder dysfunction. At multivariate logistic regression analysis, ED score significantly explained the variance of most of SCL-90-R dimension scores, whereas no association was revealed between global BI and any score of SCL-90-R dimensions. Bowel/bladder BI explained only to a very low extent the variance of depressive symptoms. CONCLUSIONS Healthcare providers should be aware of the importance of managing ED in spinal cord-injured men, as it represents a major determinant of their psychological distress, independently of the degree of FI impairment.

Dynamics of the Global Tyrosine Phosphorylation During Capacitation and Acquisition of the Ability to Fuse with Oocytes in Human Spermatozoa

Abstract Phosphorylation of tyrosine residues in cellular proteins represents a major event during sperm capacitaton, but its relationship with the acquisition of sperm-fertilizing ability is still unclear. In this study we explored the relationship between the kinetics of the global tyrosine phosphorylation, monitored with a flow cytometric assay, and the acquisition of the human sperm ability to fuse with oocytes, evaluated with the progesterone-enhanced hamster egg penetration test. Sperm tyrosine phosphorylation appeared to be an early event in the capacitation process, with a 3.6-fold mean increase within 1 h of capacitation, but at this time sperm-oocyte fusion was extremely poor compared with that observed at 5 h of capacitation. Capacitation in calcium-free medium produced a 2-fold mean increase in tyrosine phosphorylation compared with that seen in complete capacitation medium both at 1 h and 5 h of capacitation, whereas sperm-oocyte fusion significantly increased only at 1 h, remaining unchanged at 5 h of capacitation. The cAMP analog, N,2-O-dibutyryladenosine 3′,5′-cyclic monophosphate (dbcAMP), prevented the inhibitory effect of seminal plasma on tyrosine phosphorylation but not on sperm-oocyte fusion. In conclusion, these results suggest that the acquisition of sperm-fertilizing ability is always associated with an increase of the global tyrosine phosphorylation, but tyrosine phosphorylation does not necessarily reflect the acquisition of the sperm-fertilizing ability. Flow cytometry assay, a reliable technique to quickly quantify the global levels of the human sperm tyrosine phosphorylation, could be useful for a further elucidation of the biological meaning of this process, with the perspective of its clinical use as a measure of the sperm-fertilizing potential.

Energetic Metabolism and Human Sperm Motility: Impact of CB1 Receptor Activation

It has been reported that the endocannabinoid anandamide (AEA) exerts an adverse effect on human sperm motility, which has been ascribed to inhibition of mitochondrial activity. This seems to be at variance with evidence suggesting a major role of glycolysis in supplying ATP for sperm motility; furthermore, the role of AEA-binding receptors in mediating mitochondrial inhibition has not yet been explored. In this study, human sperm exposure to Met-AEA (methanandamide, nonhydrolyzable analog of AEA) in the micromolar range significantly decreased mitochondrial transmembrane potential (ΔΨm), similarly to rotenone, mitochondrial complex I inhibitor. The effect of Met-AEA (1 μm) was prevented by SR141716, CB(1) cannabinoid receptor antagonist, but not by SR144528, CB(2) antagonist, nor by iodoresiniferatoxin, vanilloid receptor antagonist. The effect of Met-AEA did not involve activation of caspase-9 or caspase-3 and was reverted by washing. In the presence of glucose, sperm exposure either to Met-AEA up to 1 μm or to rotenone for up to 18 h did not affect sperm motility. At higher doses Met-AEA produced a CB(1)-independent poisoning of spermatozoa, reducing their viability. Under glycolysis blockage, 1 μm Met-AEA, similarly to rotenone, dramatically abolished sperm motility, an effect that was prevented by SR1 and reverted by washing. In conclusion, CB(1) activation induced a nonapoptotic decrease of ΔΨm, the detrimental reflection on sperm motility of which could be revealed only under glycolysis blockage, unless very high doses of Met-AEA, producing CB(1)-independent sperm toxicity, were used. The effects of CB(1) activation reported here contribute to elucidate the relationship between energetic metabolism and human sperm motility.

Soluble products of Escherichia coli induce mitochondrial dysfunction-related sperm membrane lipid peroxidation which is prevented by lactobacilli.

Unidentified soluble factors secreted by E. coli, a frequently isolated microorganism in genitourinary infections, have been reported to inhibit mitochondrial membrane potential (ΔΨm), motility and vitality of human spermatozoa. Here we explore the mechanisms involved in the adverse impact of E. coli on sperm motility, focusing mainly on sperm mitochondrial function and possible membrane damage induced by mitochondrial-generated reactive oxygen species (ROS). Furthermore, as lactobacilli, which dominate the vaginal ecosystem of healthy women, have been shown to exert anti-oxidant protective effects on spermatozoa, we also evaluated whether soluble products from these microorganisms could protect spermatozoa against the effects of E. coli. We assessed motility (by computer-aided semen analysis), ΔΨm (with JC-1 dye by flow cytometry), mitochondrial ROS generation (with MitoSOX red dye by flow cytometry) and membrane lipid-peroxidation (with the fluorophore BODIPY C11 by flow cytometry) of sperm suspensions exposed to E. coli in the presence and in the absence of a combination of 3 selected strains of lactobacilli (L. brevis, L. salivarius, L. plantarum). A Transwell system was used to avoid direct contact between spermatozoa and microorganisms. Soluble products of E. coli induced ΔΨm loss, mitochondrial generation of ROS and membrane lipid-peroxidation, resulting in motility loss. Soluble factors of lactobacilli prevented membrane lipid-peroxidation of E. coli-exposed spermatozoa, thus preserving their motility. In conclusion, sperm motility loss by soluble products of E. coli reflects a mitochondrial dysfunction-related membrane lipid-peroxidation. Lactobacilli could protect spermatozoa in the presence of vaginal disorders, by preventing ROS-induced membrane damage.

Effect of vaginal probiotic lactobacilli on in vitro–induced sperm lipid peroxidation and its impact on sperm motility and viability

A combination of three selected strains of lactobacilli (Lactobacillus brevis [CD2], L. salivarius [FV2], and L. plantarum [FV9]), whose effectiveness in treating bacterial vaginosis in the form of vaginal tablets has been reported recently, prevented sperm lipid peroxidation that was induced in vitro by a ferrous ion promoter, thus preserving sperm motility and viability. This finding suggests the potential of vaginal probiotic lactobacilli for protecting human spermatozoa from radical oxygen species in the presence of vaginal disorders, thereby improving the fertilization potential of the female host.

Ultrasonographic determination of caput epididymis diameter is strongly predictive of obstruction in the genital tract in azoospermic men with normal serum FSH

The relationship between epididymis ultrasonography (US) and infertility is poorly defined probably owing to lack of objective and reproducible criteria of US evaluation. Here, we evaluated US size of testes, caput and of corpus epididymis in infertile men: 165 with total sperm count ≥39 × 106, 187 with total sperm count <39 × 106 and 75 azoospermic men. Blood levels of follicle stimulating hormone (FSH) and of total testosterone were also evaluated. US measures obtained using a high‐frequency (12 MHz) linear array transducer, included the mean value of bilateral testicular volumes (mL) (Testes‐M), of bilateral longitudinal diameter of caput epididymis (mm) (Caput‐M) and of the bilateral antero‐posterior diameter of the corpus measured on a longitudinal scan (mm) (Corpus‐M). Testicular histology of azoospermic men was obtained and the percentage of seminiferous tubules with elongated spermatids (%T) was used to classify cases with normal spermatogenesis (obstructive azoospermia) (n = 17; %T ≥ 80), or with deranged spermatogenesis (n = 58; %T ≤ 33). Caput‐M was correlated with Testes‐M (p = 0.0003; r = 0.17) and with FSH serum levels (p = 0.024; r = −0.14) but not with semen parameters. Caput‐M but not Corpus‐M values resulted greater in obstructive azoospermia compared with other groups, but difference was not significant. Cut‐off values of Testes‐M, Caput‐M and of FSH correctly classified cases of obstructive azoospermia (AUC > 0.5). A patient with FSH < 7.8 IU/mL had a 63.6% chance (CI 40.1–83.2%) of being affected by obstructive azoospermia. US Caput‐M ≥10.85 mm, which represented the cut‐off value with the highest combination of sensitivity (58.8%, CI 32.9–81.6%) and specificity (91.4%, CI 81.0–97.1%) applied in cases with FSH < 7.8 IU/mL increased the probability for obstructive azoospermia from 63.6% up to 92.3% (CI 76.5–98.8%). US evaluation of the caput epididymis diameter helped in predicting the obstructive origin of azoospermia when FSH was not increased, whereas it was not relevant in non‐azoospermic men.

Involvement of mitochondrial dysfunction in the adverse effect exerted by seminal plasma from men with spinal cord injury on sperm motility

The aetiology of severe asthenozoospermia in men with spinal cord injury includes an adverse impact of seminal plasma (SP) on sperm motility. In this study we investigated the effect exerted by SP from men with SCI on donor sperm mitochondrial activity and its reflection on motility. Donor spermatozoa were exposed (1 h) to SP from 22 ejaculates of men with SCI. Only SP from samples exhibiting both a low fructose level and an inhibitory effect on mitochondrial membrane potential (ΔΨm), assessed at flow cytometry with JC‐1, affected donor sperm motility when evaluated 1 h after co‐incubation. This effect was reverted by washing from SP and sperm re‐suspension in medium containing glucose, in spite of persistently depressed ΔΨm. In the same samples, sperm motility and vitality dramatically decreased when evaluated 6 h after washing and re‐suspension in the glucose‐containing medium. Seminal plasmas which induced a disruption of ΔΨm, also enhanced a mitochondrial ROS generation, as assessed by MitoSOX red. The enhanced mitochondrial ROS generation was associated with a late induction of sperm membrane lipid peroxidation, as assessed by BODIPY C11, when evaluated at 6 h, but not at 1 h, after washing from SP. Furthermore, activation of caspase‐9 and caspase‐3 accompanied the loss of ΔΨm. In conclusion, a double energetic blockage (glycolysis and mitochondrial respiration) can represent a metabolic determinant of the early adverse effect exerted by SP from men with SCI on sperm motility. Mitochondrial dysfunction‐related oxidative/apoptotic mechanisms can account for later consequences on sperm motility/vitality.

Intrauterine insemination with or without mild ovarian stimulation in couples with male subfertility due to oligo/astheno- and/or teratozoospermia or antisperm antibodies: a prospective cross-over trial.

Seventy-three couples with male subfertility, which was due to oligo/astheno- and/or teratozoospermia (n = 63) or antisperm antibodies (n = 10), were randomly assigned to sequential timed natural intercourse, intrauterine insemination (IUI) and IUI + mild ovarian hyperstimulation. From the analysis of 384 observed cycles, IUI was shown to be effective in oligo/asthenozoospermia without severe teratozoospermia, when it was associated with moderate multifollicular induction, and in male immunologic subfertility, IUI was highly effective in nonstimulated cycles also.

RANTES and human sperm fertilizing ability: effect on acrosome reaction and sperm/oocyte fusion.

Beta-chemokine, regulated on activation and normally T-cell expressed and presumably secreted (RANTES), is present in both the male and female genital tract fluids where its levels increase in diseases related to infertility, such as endometriosis and male genital tract infections. beta-Chemokine receptors (CCR3 and CCR5) are expressed on freshly ejaculated human sperm cells and a sperm chemoattractant effect for RANTES has been reported. No information exists on other possible roles of RANTES on sperm functions involved in the fertilization process. In the present study, the exposure of sperm suspensions to high concentrations of the chemokine, comparable to those observed in inflammatory diseases, significantly decreased the stimulatory effect exerted by progesterone on sperm/oocyte fusion, evaluated by means of the hamster egg penetration test. Accordingly, a large proportion of spermatozoa preincubated under capacitating conditions with high concentrations of RANTES underwent a premature acrosome reaction (AR) that prevented subsequent progesterone-induced AR. Finally, sperm samples exposed to the same high levels of chemokine showed a significant increase in the intracellular levels of cAMP, which is involved in capacitation and AR dynamics. These results indicate a negative interference of high levels of RANTES on the sperm fertilizing ability, thereby suggesting a potential contribution of this chemokine to subfertility associated with endometriosis and genital tract inflammatory diseases.