Seasonal trends in the prevalence of hypospadias: Aetiological implications.

The aim of the present study was to examine the seasonality of hypospadias in Greece in an attempt to elucidate the aetiology. All boys born between 1991-1998, who underwent hypospadias repair at 'Aghia Sophia' Children's Hospital, Athens (n=542) were analysed. All Greek live-born males during the same period (population at risk; m=421,175) served as the controls. Seasonality by month of birth was evaluated with specific statistical tools. Meteorological parameters were also analysed. All tests yielded significant results, suggesting a simple harmonic prevalence pattern (highest/lowest: autumn, peak in October/spring, trough in April). Therefore, the first trimester of hypospadiac gestations coincides more frequently with winter. Meteorological parameters varied seasonally (maximal sunlight; air temperature in summer/minimal in winter, maximal rainfall in winter/minimal in summer) and were strongly associated pairwise. Hypospadiac birth prevalence follows a simple harmonic seasonal pattern and is associated with that of cryptorchidism in Greece. The coincidence of the first or third trimester of a potentially genetically influenced gestation with winter could lead to the phenotypic expression of hypospadias or cryptorchidism, respectively. The potential role of a cyclic-varied androgen-production stimulator, such as human chorionic gonadotrophin may be speculated. The seasonality of a common environmental factor acting directly/indirectly may contribute to these patterns, and possibly to the common pathogenesis of these congenital malformations.

Genetic analysis of the human Insulin-like 3 gene: absence of mutations in a Greek paediatric cohort with testicular maldescent.

This study investigated the hypothesis that genetic alterations of the human insulin-like 3 (INSL3) gene are associated with testicular maldescent (TMD). Genomic DNA was extracted and amplified from peripheral blood samples of 170 unrelated children with all possible phenotypical expressions of TMD and 50 volunteers with normal external genitalia from the general paediatric population (controls). PCR-single strand conformation polymorphism analysis was used to screen INSL3 gene for genetic variants. For rapid screening of a detected nonsilent genetic alteration, restriction assay using endonuclease Eag I was further employed. Products were analysed on 2% agarose gel and restriction patterns were visualised by ethidium bromide. Differences in genotype and allelic distributions of nonsilent genetic alterations were evaluated between (i) patients-controls, (ii) familial-sporadic, (iii) bilateral-unilateral and (iv) intra-abdominal-inguinal cases of TMD. No mutations were detected. Three common INSL3 gene polymorphisms (27G>A, 126G>A, 178G>A) unrelated to any particular phenotype of TMD were detected both in patients and controls. These results indicate that INSL3 gene mutations are not a common cause of TMD in the human.

The (TAAAA)n polymorphism of sex hormone-binding globulin gene is not associated with testicular maldescent.

The aim of this family-based study was to investigate the potential association/genetic linkage of the (TAAAA)n polymorphism of sex hormone-binding globulin gene proximal promoter with testicular maldescent (TMD). Genomic DNA was extracted from the peripheral blood of 487 subjects (174 index families): (i) 180 children with all phenotypes of TMD, (ii) 307 parents (156 mothers and 151 fathers). Conventional polymerase chain reaction amplification products were electrophoresed on 10% nondenaturating polyacrylamide gel and visualised by silver staining. After excluding ambiguous parental-child trios and most cases of index families with missing parental genotypes, 429 individuals were left for analysis: 138 completely typed nuclear families (five included a second affected child) and five child-parent couples (one parent missing). Eight fathers presented history of TMD, that is, a total of 156 cases with TMD were analysed. Alleles were analysed with the affected family-based control method and logistic regression-based extension of the transmission disequilibrium test for multiallelic loci. (ΤΑΑΑΑ)n polymorphism analysis revealed six alleles based on repeat numbers (n=5-10). No association/genetic linkage between the (TAAAA)n polymorphism and TMD was detected. Other factors should be investigated to potentially explain the genetic predisposition that seems to exist in at least a subgroup of these patients.

Screening for Y chromosome microdeletions in childhood: lack of evidence for a direct association with testicular maldescent.

The aim of the study was to investigate the hypothesis that Y chromosome microdeletions are directly implicated in testicular maldescent. Genomic DNA was extracted from the peripheral blood of 292 subjects. This population consisted of (i) 180 children with all phenotypes of isolated (non-syndromic) testicular maldescent from 174 index families, (ii) affected adult relatives available (n = 12) and (iii) 100 unrelated children with normal external genitalia (controls). The sequence-tagged site primer set and the conditions of conventional polymerase chain reaction amplification were based on the current laboratory guidelines for molecular diagnosis of Y chromosome microdeletions recommended by the European Academy of Andrology and the European Molecular Genetics Quality Network. Two multiplex reactions were designed to screen the regions of azoospermic factors a, b and c. Each multiplex reaction included adequate internal and external amplification controls. Amplification products were submitted to electrophoresis on 2% agarose gel impregnated with ethidium bromide dye solution for 80 volt-h and visualised under ultraviolet light. No microdeletions were detected in any subject. These results indicate that Y chromosome microdeletions are not directly implicated in the pathogenesis of testicular maldescent. Other factors should be investigated to potentially explain the genetic predisposition that seems to exist in at least a subgroup of these patients.

Comparison of angiogenic activity after urethral reconstruction using free grafts and pedicle flap: an experimental study.

PURPOSE: An experimental study was undertaken in order to estimate the angiogenic activity in different free grafts and pedicle flap in urethral reconstruction in an animal model. METHODS: Twenty-eight white New Zealand rabbits were randomly divided into five groups (O, A, B, C and D). A ventral urethral defect was created in all groups. In the group O, (n = 4), a simple closure of the defect was performed. Free penile skin graft (group A, n = 6), buccal mucosal graft (group B, n = 6), bladder mucosal graft (group C, n = 6), and pedicle penile skin graft (group D, n = 6) were used to bridge the urethral defect as an onlay patch. The animals were euthanized on the 21st postoperative day. The angiogenic activity was assessed with immunohistochemistry, using the anti-CD31 MoAb and the alkaline phosphatase antialkaline phosphatase procedure. The native vascularity of penile skin as well as buccal and bladder mucosa was assessed in rabbits from group O (n = 3). Statistical analysis was performed using one-way ANOVA. RESULTS: The angiogenesis seen with a magnification of x 200 in groups O, A, B, C, and D was 34.1 +/- 4.1 (mean +/- SD), 61.7 +/- 6.4, 94.3 +/- 6.4, 91.5 +/- 7.2, and 30.8 +/- 5.2 vessels per optical field, respectively. There were statistically significant differences (p < 0.001) between group O and groups A, B, C and between group A and groups B, C, D, but not (p > 0.5) between groups B and C and groups O and D. The native vascularity of penile skin, buccal mucosa and bladder mucosa was 23.3 +/- 3.0, 24.6 +/- 3.7 and 17.0 +/- 2.6 vessels per optical field, respectively. CONCLUSION: Buccal and bladder mucosal grafts exhibit a higher angiogenic activity than free and pedicle penile skin flap when transplanted in urethral defects. As the buccal mucosal graft showed the higher angiogenic activity and its harvesting is easier than bladder mucosa, we propose that in urethral reconstruction surgery the use of this graft might offer more reliable results.

Annular pancreas in two consecutive siblings: an extremely rare case.

Annular pancreas is the rare congenital anomaly where the pancreas forms a full or incomplete ring around the second segment of the duodenum, causing various degrees of stenosis or atresia. It is estimated that it appears in 1 out of 12 000-15 000 births of living neonates and until now, in the literature, only 6 cases have been reported among individuals of the same family. We present the case of two siblings, a boy and a girl, with annular pancreas from consecutive pregnancies of the same couple. Both neonates had a prenatal diagnosis of duodenal obstruction and they underwent duodenoduodenal, proximal transverse to distal longitudinal anastomosis. Furthermore, the girl had a mobile ascending colon. Their postoperative condition was perfect. The case we are reporting is an addition to the other 6 cases of familial presentation of annular pancreas and is similar to one of them. In these families, a total of 16 persons present this congenital anomaly while 14 are seemingly healthy. Twelve of the affected persons are female and 4 male. In conclusion, it can be stated that female individuals seem to have a greater propensity to transmit the disease to their descendants, compared to males, suggesting the possible action of an autosomal recessive sex-influenced gene. The recording of such rare family cases should be encouraged, in order to fully recognize a possible type of inherited transmission.

Colostomy type in anorectal malformations: 10-years experience.

AIM: The aim of this study was to evaluate the influence of colostomy type on morbidity during the treatment of anorectal malformations. METHODS: Sixty-eight infants (male: female ratio 1.3:1) with anorectal malformations that required colostomy were treated in our clinics during the period 1991-2001. Of these patients, 26 had received a loop colostomy: 14 of these underwent posterior sagittal anorectoplasty (PSARP) at the age of 9-12 months (Group A), and 12 underwent PSARP at the age of 2-4 months (Group B). Forty-two infants received a separated-stomas colostomy and underwent PSARP at the age of 9-12 months (Group C). The incidence of complications among groups was compared using the 2 sided Fisher's exact test. RESULTS: Eight cases from group A were complicated with prolapse of the stomas, perianal wound infection, pull-through dehiscence, and anal fibrotic stricture. The only complication observed in groups B and C was perianal wound infection, which occurred in 1 case from each group. A statistically significant difference was observed in the incidence of complication between groups A and C (p<0.001) and between groups A and B (p=0.014). The results from groups B and C did not differ significantly (p=0.398). When the cases complicated with colostomy prolapse were removed from the statistical analysis, groups A and C still differed significantly (p=0.001) but groups A and B did not (p=0.069). CONCLUSIONS: As the incidence of complications increases with time after a loop colostomy, we encourage either an early corrective procedure or the modification into separated-stomas colostomy (SSC) before PSARP is performed for those cases that would involve definitive surgery in late infancy.

Spontaneous pneumomediastinum: is a chest X-ray sufficient?

Spontaneous pneumomediastinum (SPM) is an uncommon disease defined as a non-traumatic presence of free air in the mediastinum, without underlying disease. We present a 13-year- old boy who was previously in a perfect health, who was presented with subcutaneous cervical emphysema, dysphagia, chest and neck pain. The chest roentgenogram revealed the presence of subcutaneous emphysema without any other abnormal findings. A computer tomography (CT) scan was obtained, and this confirmed the existence of subcutaneous cervical emphysema and also the presence of pneumomediastinum. The child's progress was uneventful and after 6 days he was discharged from the hospital in excellent clinical condition. We propose that chest CT is useful, in less obvious cases of SPM, to detect the free air in the mediastinum and probably SPM is underdiagnosed in clinical practice in the young people.

Cryptorchidism: seasonal variations in Greece do not support the theory of light.

To examine seasonal trends of cryptorchidism in Greece, 583 males with true isolated cryptorchidism were analyzed. All 208 912 live-born boys born during the same period were used as a comparison group. Seasonality by month of birth was evaluated using both Edwards' model with adjusted frequencies and exact theta(i), and Walter-Elwood method with exact theta(i). Both tests resulted in consistent findings. The incidence of cryptorchid births in Greece follows a documented cyclic pattern of simple harmonic type with spring being the season of statistical predominance (peak in March with a second, almost equivalent, peak in May). In contrast, in autumn the incidence of cryptorchid births was considerably lower (trough in September). Given the fact that no significant differences in daylight length are found among seasons in Greece, the detection of a significant seasonal variation suggests that factors other than light are involved in the pathogenesis of cryptorchidism. Low environmental temperature is proposed as a causative factor negatively influencing the maternal hCG profiles and the inguinoscrotal phase of testicular descent. This is further supported by: (i) the similarity of our results to those reported by other European countries of different longitude and geographical width and (ii) our data showing significantly smaller maternal hCG profiles at the 26th week of gestation during winter compared with summer.

Sonographic features indicative of hydrostatic reducibility of intestinal intussusception in infancy and early childhood.

The aim of the study was to investigate the various sonographic patterns of intussusception, which may be indicative of its reducibility by hydrostatic reduction. Seventy-one infants and children clinically suspected of having intussusception were studied sonographically and given barium enemas. Sonographic patterns of intussusception were correlated with its reducibility. When the head of intussusception appeared as a target-like mass the hydrostatic reduction rate was 100%. When demonstrated as a doughnut-like mass, the intussusception reducibility depended on the thickness of the hypoechoic external ring of the "doughnut". With a thickness equal to, or less than, 7.2 mm the reduction rate was 100%, with a thickness ranged between 7.5 and 11.2 mm the reduction rate was 68.9% and with a thickness measured 14 to 14.2 mm a surgical resection of bowel was required. When a small amount of fluid appeared within the head of intussusception, like a crescent hypoechoic area, the hydrostatic reduction was unsuccessful, regardless of the sonographic pattern of intussusception (target- or doughnut-like mass). The presence of a small amount of free peritoneal fluid did not affect the reducibility of intussusception. In conclusion, sonographic patterns of intussusception identify good and poor prognostic features for the success of hydrostatic reduction.

Ooplasmic injection of elongating spermatids for the treatment of non-obstructive azoospermia.

We applied the technique of ooplasmic elongating spermatid injection to the treatment of non-obstructive azoospermia. Mature oocytes were injected with elongating spermatids isolated from testicular biopsy material obtained from 13 non-obstructed azoospermic men. Seventy-three oocytes were successfully injected with elongating spermatids and were then cultured for 36 h. At 13 h post-injection 68 oocytes were found to be activated and 52 of them were fertilized. Forty-one 2- to 4-cell stage embryos developed from normally fertilized oocytes were transferred. At least two embryos were transferred to each female partner. Two pregnancies were achieved. Elongating spermatid injection may have a role in the treatment of non-obstructive azoospermia.

Effects of seminal plasma from cigarette smokers on sperm viability and longevity.

OBJECTIVE: To evaluate the effects of cigarette smoking on the ability of seminal plasma (SP) to maintain sperm viability. DESIGN: Clinical randomized study. Spermatozoa from cigarette smoking or nonsmoking subjects were reconstituted in SP from smokers and nonsmokers and in modified Ham's F-10 medium, followed by sperm quality assessment during a 48-hour incubation period. SETTING: Andrology Institute of Lexington, Lexington, Kentucky. PATIENT(S): Twenty men who had been smoking cigarettes for longer than 3 years (30 cigarettes per day or more) and 20 nonsmokers participated in this study. MAIN OUTCOME MEASURE(S): Improvement in sperm viability by removal of SP--and associated detrimental factors present in the SP--from smoker subjects. RESULT(S): The results obtained indicate that the quality of spermatozoa obtained from nonsmokers was superior to that of smokers. The SP from the two patient groups had a definite effect on their respective sperm quality, i.e., beneficial effects for the nonsmokers, detrimental effects for the smokers. Exposure of spermatozoa from the nonsmokers to SP from the smokers resulted in a significant reduction in sperm viability. However, exposure of spermatozoa from the smokers to SP from the nonsmokers or to Ham's F-10 medium yielded significant improvements in sperm viability. CONCLUSION(S): The detrimental effects of smokers' SP on nonsmokers' spermatozoa was prominent and a rather unique phenomenon. The results generated in this study could be of clinical significance since removal of smokers' SP and subsequent reconstitution and incubation in physiological media seems to enhance the viability, longevity, and possibly the fertilizing ability of these spermatozoa for use in various assisted reproductive technologies.

Ooplasmic round spermatid nuclear injection procedures as an experimental treatment for nonobstructive azoospermia.

PURPOSE: Our objective was to apply ooplasmic round spermatid nuclear injections for the treatment of nonobstructive azoospermia. MATERIALS: Participants were nine azoospermic men who had previously undergone diagnostic testicular biopsy. Spermatogenetic arrest was diagnosed at the round spermatid stage (n = 6) or primary spermatocyte stage (n = 3). A second (therapeutic) testicular biopsy was performed and round spermatid nuclei were recovered from all the participants. RESULTS: Forty-nine mature oocytes were successfully injected with nuclei and then cultured for 72 hr. Twenty-four embryos were transferred to nine women. No pregnancy was achieved. CONCLUSIONS: Round spermatids can be recovered from therapeutic testicular biopsy material of men negative for round spermatids in previous routine diagnostic testicular biopsy specimens. Round spermatid nuclear injections may play a role in the treatment of nonobstructive azoospermia.

Bilateral effect of unilateral vasectomy on testicular testosterone biosynthesis.

Twelve immature male dogs underwent a left vasectomy (group A). An additional five underwent a sham operation (group B). Sixteen weeks after the surgery, the bilateral mean values for caudal epididymal sperm content, the percentage of motile spermatozoa, intratesticular testosterone concentration, and testicular secretion of androgen-binding protein (in vitro) were significantly lower in group A. The mean peripheral serum testosterone responses 3 hours after human chorionic gonadotropin stimulation (3,000 IU) were significantly lower in group A than in group B (6.3 ng/mL v 9.5 ng/mL). These findings indicate a bilateral deficiency in both Leydig and Sertoli cell secretory function in unilaterally vasectomized dogs, resulting in impaired bilateral spermatogenesis and sperm maturation. The authors suggest that unilateral injuries of the vas deferens during hernia operations in children may result in bilateral testicular dysfunction.