Basal and post-ischemic vascular compliance in children/adolescents born small for gestational age.

BACKGROUND: Intrauterine growth restriction plays a powerful role in influencing later susceptibility to certain chronic diseases, such as hypertension. Endothelial dysfunction and arterial stiffness are early events in the development of cardiovascular diseases (CVDs). We have studied vascular compliance in small for gestational age (SGA) children/adolescents in comparison with that in appropriate for gestational age (AGA) subjects. METHODS: We monitored blood pressure, vascular resistance and compliance in 82 children-adolescents (52 SGA, 30 AGA), by means of pulse wave analysis (CR 2000 HDI) at the radial level, before and after 3 min of ischemic stress at the brachial level. RESULTS: In the children/adolescents born SGA we found a significant increase in systolic and diastolic blood pressure and vascular resistance in the basal condition; the large and small vessels were stiffer. After ischemia we observed an increased vascular response in the SGA children/adolescents: there was a great diminution of systolic and diastolic blood pressure and a larger increase of the elasticity of the conduit and resistance vessels. CONCLUSIONS: These data show that the SGA group presented some early signs of arterial wall functional disorders. More pediatric data are needed for the evaluation by non-invasive techniques of vascular function in children-adolescents at risk of CVD.

Distribution of α- and δ-tocopherols in seminal plasma and sperm fractions of men with normal and abnormal semen parameters.

The aim of this study was to investigate the presence of the different isoforms of tocopherol (T) in seminal plasma (P) and in the sperm fractions of individuals with abnormal (group 1) and normal (group 2) sperm parameters; the relationships between these isoforms and conventional sperm parameters were also explored. Two vitamin E homologues, α-T and δ-T, were identified in the semen of all participants. Although α-T and δ-T concentrations were similar in the semen of the 2 groups, group 1 showed a lower α-T ratio (S/P) (0.90 vs. 1.20, P < .001) and δ-T ratio (0.86 vs 1.13, P = .007) than group 2. In addition, both T ratios were correlated with the percentage of viable cells, detected by eosin staining. These results suggested that α-T and δ-T are not homogeneously distributed in the semen fractions; in normal semen they are more concentrated in the sperm membrane, whereas in abnormal semen the damaged sperm cells may release both Ts in the plasma. To verify whether sperm membrane breakage could alter α-T and δ-T distribution between the seminal plasma and the spermatozoa, normal sperm samples were sonicated; after sonication a consistent sperm plasma membrane fragmentation, highlighted by transmission electron microscopy, and a concomitant release of α-T and δ-T were observed. In conclusion, the Ts coupled directly with the sperm membrane seem to play the main protective role in the semen, and the release of α-T and δ-T in the P fraction is probably an index of lower antioxidant power and sperm quality.

Early identification of the risk for free radical-related diseases in preterm newborns.

BACKGROUND: Despite recent advances in preterm newborns healthcare, perinatal pathologies and disabilities are increasing. Oxidative stress (OS) is determinant for the onset of an unbalance between free radicals (FRs) production and antioxidant systems which plays a key role in pathogenesis of pathologies such as retinopathy of prematurity (ROP), bronchopulmonary dysplasia (BPD), necrotizing enterocolitis (NEC), intraventricular hemorrhage (IVH), grouped as 'free radical-related diseases' (FRD). AIM: This study tests the hypothesis that OS markers levels in cord blood may predict the onset of FRD pathologies. PATIENTS AND METHODS: 168 preterm newborns of GA: 24-32weeks (28.09+/-1.99); and BW: 470-2480 gr (1358.11+/-454.09) were consecutively recruited. Markers of potential OS risk (non-protein bound iron, NPBI; basal superoxide anion, BSA; under stimulation superoxide anion, USSA) and markers of OS-related damage (total hydroperoxides, TH; advanced oxidation protein products, AOPP) were assessed in cord blood. Associations between FRD onset and OS markers were checked through inferential analysis (univariate logistic regression). RESULTS: The development of FRD was significantly associated to high cord blood levels of TH, AOPP and NPBI (respectively p=0.000, OR=1.025, 95%CI=1.013-1.038; p=0.014, OR=1.092, 95%CI=1.018-1.172; p=0.007, OR=1.26995%CI=1.066-1.511). CONCLUSIONS: Elevated levels of TH, AOPP and, above all, NPBI, in cord blood are associated with increased risk for FRD. OS markers allow the early identification of infants at risk for FRD because of perinatal oxidant exposure. This can be useful in devising strategies to prevent or ameliorate perinatal outcome.

Birefringence characteristics in sperm heads allow for the selection of reacted spermatozoa for intracytoplasmic sperm injection.

OBJECTIVE: To verify clinical outcome after injection of spermatozoa that have undergone the acrosome reaction (reacted spermatozoa) vs. those still having an intact acrosome (nonreacted spermatozoa). DESIGN: Prospective, randomized study. SETTING: Reproductive Medicine Unit, Italian Society for the Study of Reproductive Medicine, Bologna, Italy. PATIENT(S): According to a prospective randomization including 71 couples with severe male factor infertility, intracytoplasmic sperm injection (ICSI) was performed under polarized light that permitted analysis of the pattern of birefringence in the sperm head. Twenty-three patients had their oocytes injected with reacted spermatozoa, 26 patient's oocytes were injected with nonreacted spermatozoa, and in 22 patients both reacted and nonreacted spermatozoa were injected. INTERVENTION(S): Intracytoplasmic sperm injection was performed under polarized light to selectively inject acrosome-reacted and acrosome-nonreacted spermatozoa. MAIN OUTCOME MEASURE(S): Rates of fertilization, cleavage, pregnancy, implantation, and ongoing implantation. RESULT(S): There was no effect on the fertilizing capacity and embryo development of either type of sperm, whereas the implantation rate was higher in oocytes injected with reacted spermatozoa (39.0%) vs. those injected with nonreacted spermatozoa (8.6%). The implantation rate was 24.4% in the group injected with both reacted and nonreacted spermatozoa. The delivery rate per cycle followed the same trend. CONCLUSION(S): Spermatozoa that have undergone the acrosome reaction seem to be more prone to supporting the development of viable ICSI embryos.

Infection by CagA-positive Helicobacter pylori strains may contribute to alter the sperm quality of men with fertility disorders and increase the systemic levels of TNF-alpha.

This study was aimed to address the possibility that Helicobacter pylori infection may play a detrimental role in semen quality of men with idiopathic infertility. Infection by H. pylori and by strains expressing CagA was determined in 80 male infertile patients by Western blotting and ELISA. Semen analysis was performed by light microscopy and transmission electron microscopy quantitatively elaborated (fertility index, immaturity, necrosis, and apoptosis percentages). Systemic levels of IL-6 and TNF-alpha were evaluated. Infertile patients infected with H. pylori showed a low sperm quality respective to uninfected patients. Particularly, in CagA-positive patients we observed a significant reduction in sperm motility and in the fertility index, while apoptosis and necrosis were increased. In these patients, the means of systemic TNF-alpha levels were higher than those of uninfected patients. The negative influence of CagA-positive H. pylori infection on sperm quality may help to understand the role of chronic infections in reproductive disorders.

The presence of bacteria species in semen and sperm quality.

PURPOSE: To verify the prevalence of semen bacterial contamination and whether the contamination could decrease sperm quality. METHODS: Spermiogram, semen culture, and sperm transmission electron microscopy (TEM) analysis were performed. TEM data were elaborated using a mathematical formula that calculates a fertility index (FI)--able to define patients as fertile or infertile--and the percentage of sperm apoptosis, immaturity and necrosis. We aligned the amino acid sequence of beta-tubulin with protein of the most frequent species isolated from semen. RESULTS: Patients were divided according to the contaminating species; in each group, we observed fertile individuals, in whom the semen quality was similar to that of controls and infertile men whose sperm quality was significantly decreased, in terms of motility, FI, apoptosis and necrosis. Partial homology between beta-tubulin and bacterial proteins was observed. CONCLUSION: Sperm bacterial contamination is quite frequent and could contribute to the deterioration of the sperm quality of infertile men.

Effect of emotional stress on sperm quality.

BACKGROUND & OBJECTIVE: Emotional stress plays a detrimental role on fertility. In this study male patients with idiopathic infertility were selected after evaluation of psychological stress to evaluate a positive effect of a stress therapy on their semen quality. METHODS: A total of 20 patients with infertility were enrolled in the study and randomly divided in two groups. Ejaculates were examined by light and transmission electron microscopy (TEM). Meiotic segregation was also investigated by fluorescence in situ hybridization (FISH). Ten patients were treated with Conveyer of Modulating Radiance (CRM) therapy and sperm characteristics and meiotic segregation were evaluated again three months at the end of treatment. RESULTS: TEM data showed that, among sperm pathologies, necrosis and apoptosis were higher and the number of "healthy" sperm was significantly reduced in both groups of stressed men compared to reference values. The number of "healthy" sperm was significantly higher in the treated group after therapy, indicating a recovery of sperm quality, although no significant decrease in sperm pathologies was observed. FISH analysis showed that the mean frequencies of sex chromosomes disomies and diploidies significantly decreased after stress therapy. INTERPRETATION & CONCLUSION: The effects induced by stress also seem to include meiotic and structural alterations in sperm cells. The spermatogenic process was improved after a cycle of CRM therapy indicating that stress is an additional risk factor for idiopathic infertility.

Microsurgery for recurrent lumbar disk herniation at the same level and side: do patients fare worse? Experience with 95 consecutive cases.

BACKGROUND: In reviewing our experience with reoperation of RLDH, our aim was mainly to determine whether patients fared worse than after primary surgery. We found no uniform answers to this question in the literature. METHODS: The data of 95 patients (29 women and 66 men) who underwent reoperation for RLDH at the same level and side were analyzed retrospectively. Forty-two patients underwent the first operation in our clinic (recurrence rate, 2.6% of 1586 cases). Gadolinium-enhanced MRI was performed in all patients. Main clinical data of patients, pain-free interval, operation time, surgical complications, duration of hospital stay, and clinical improvement rate were recorded. RESULTS: The mean pain-free interval was 55 months (range, 3-120 months). Levels of recurrent herniation were L4 through L5 and L5 through S1 (65% and 35% of cases, respectively). Revision surgery lasted longer on average than the previous diskectomy (P < .01) and was complicated by dural tear in 4 cases (4.2% vs 0.9% during primary diskectomy, P < .05). There were no significant differences between revision and previous surgery in terms of hospital stay. However, rates of excellent/good outcomes were significantly less for RLDH (89% vs 95%, P < .05); and the percentage of poor results was higher (2% vs 0.5%, P < .05). Age, sex, smoking, profession, trauma, level and degree of herniation, and pain-free interval were not correlated with clinical outcome. CONCLUSION: Conventional microsurgery for RLDH showed lightly but significantly worse results than those of primary microdiskectomy. Patients contemplating reoperation should be informed of this fact and of the risk of dural tear and prolonged operation time.

Cryptorchidism and semen quality: a TEM and molecular study.

Cryptorchidism is a pathological condition defined as the failure of the testis to descend into the scrotum, the location of the cryptorchid testis can be in the inguinal canal or in the prescrotal and abdominal area, sometimes resulting in atrophic seminiferous tubules. The aim of this study was to analyze semen quality of men who underwent orchidopexy for unilateral or bilateral cryptorchidism during childhood. Semen quality was investigated by light microscopy to evaluate sperm concentration and motility. Sperm morphology was performed by transmission electron microscope (TEM), and the data were mathematically elaborated. The presence of Y microdeletions was investigated by polymerase chain reaction. The effect of cryptorchidism on meiosis was explored by fluorescence in situ hybridization (FISH). The incidence of azoospermia was higher in the group with bilateral compared with unilateral cryptorchidism, and semen parameters were better in the unilateral group. Sperm pathologies detected by TEM indicated a severe deterioration of sperm quality in both groups. Necrosis and apoptosis appeared to be the most frequent pathologies, and their values reached statistical significance compared with those from fertile controls. The presence of chromosome Y microdeletions in patients with cryptorchidism and severe spermatogenetic defects is controversial. No microdeletions were found in this study. FISH values indicated that the mean percentage of gonosome disomies and diploidies were generally out of normal range, indicating a severe disturbance of meiotic segregation. The effects induced by cryptorchidism resolved in childhood seem to include a spermatogenetic impairment, leading to recommendation of detailed ultrastructural and chromosomal sperm analyses before undertaking assisted reproductive techniques.

Studies on varicocele III: ultrastructural sperm evaluation and 18, X and Y aneuploidies.

The idea that varicocele plays a detrimental role in fertility is supported by the presence of a higher frequency of affected men among the infertile population than among men with normal semen parameters. In this research we examined ejaculates from a large group of selected men affected by varicocele by light and electron microscopy. The effect of varicocele on chromosome meiotic segregation was investigated by fluorescence in situ hybridization (FISH). The potential benefits of varicocelectomy on sperm quality were evaluated by analyzing sperm characteristics before and after surgical correction of varicocele. Transmission electron microscopy (TEM) analysis, elaborated previously, showed that the incidence of immaturity, apoptosis, and necrosis was higher in the varicocele group than in controls. FISH analysis performed on sperm nuclei from selected patients with varicocele showed that the mean frequencies of disomies and diploidies were generally out of the normal range, indicating a severe disturbance in meiotic segregation. Sperm characteristics evaluated before and after varicocele repair showed a general improvement. As a consequence, the varicocele seem to affect sperm morphology and function concomitantly with meiotic segregation derangement. In consideration of these data, we suggest that TEM and FISH analyses should be performed for all varicocele patients.

Infertile spermatozoa in a human carrier of robertsonian translocation 14;22.

OBJECTIVE: To present the ultrastructural, functional, and chromosomal analyses of spermatozoa from an infertile man with normal phenotype and chromosomal translocation 14;22. DESIGN: Case report. SETTING: Regional Reference Center for Male Infertility in Siena, Italy. PATIENT(S): A 36-year-old man with primary infertility for 3 years and his parents. INTERVENTION(S): Family history and lymphocytic karyotypes, physical and hormonal assays, and semen analysis. MAIN OUTCOME MEASURE(S): Morphological sperm evaluation was performed by light, fluorescent, and electron microscopy; chromosomal constitution was examined by the fluorescence in situ hybridization (FISH) technique. The penetration ability of spermatozoa was checked by the hamster test. RESULT(S): The spermatozoa of the patient showed unusual ultrastructural defects. The nuclei were large, spheroidal, and generally uncondensed; the acrosomes were frequently absent or reduced; and the axonemes were often devoid of dynein arms or central singlet tubules. These characteristics are related to immaturity. The lymphocytic karyotype revealed a robertsonian translocation 14;22 in the sterile patient and his mother. FISH sperm analysis demonstrated a high frequency of diploidy for the chromosome 18,XY. The hamster penetration test gave negative results. CONCLUSION(S): The unusual structural sperm immaturity is associated with the translocation 14;22. This chromosomal anomaly may therefore negatively influence the spermatogenesis; an interchromosomal effect on meiosis segregation is also suggested.

Insulin-dependent diabetes in men is associated with hypothalamo-pituitary derangement and with impairment in semen quality.

BACKGROUND: The objective of the study was to investigate the hypothalamo-pituitary-testicular axis and sperm structure at the transmission electron microscope (TEM) level in men affected by insulin-dependent diabetes. METHODS: Twenty-two diabetic men and 24 controls were recruited. GnRH (100 micro g) was administered and FSH- and LH-induced secretion was evaluated. Semen samples were collected and sperm concentration and motility were determined using a Makler chamber. Ejaculated sperm were fixed and observed with a TEM. RESULTS: The response of gonadotrophins to GnRH was significantly lower in diabetics than in control men. Sperm motility was also significantly lower. At the electron microscope level, sperm from diabetics exhibited a higher percentage of immaturity- and apoptosis-related defects than sperm from controls. CONCLUSIONS: The reduced response of gonadotrophins to GnRH in diabetic men may indicate a decreased acute releasable pool of pituitary gonadotrophins. The results of TEM examination showed that sperm from men with diabetes presented severe structural defects in comparison with sperm from controls. It is possible that the reproductive impairment recognized in men with diabetes could be the result of interference by the disease on the hypothalamo-pituitary-testicular axis at multiple levels, as indicated by the reduced gonadotrophin response to appropriate stimuli and by the abnormal ultrastructure of ejaculated sperm. The defective spermatogenesis may be the consequence of a direct testicular effect of the disease.