Gene expression alterations in cryptorchid males using spermatozoal microarray analysis.

OBJECTIVE: To identify differential spermatozoal gene expression patterns using microarray between controls and patients with cryptorchidism. DESIGN: Prospective laboratory analysis. SETTING: Pediatric urology clinic and laboratory of an academic hospital in the United States. PATIENT(S): Ten control patients and 12 cryptorchid males (8 unilateral, 4 bilateral). INTERVENTION(S): Ejaculates were collected and motile sperm were isolated by Percoll centrifugation, total RNA was extracted and verified using sperm-specific reverse transcriptase-polymerase chain reaction (RT-PCR). Biotin-labeled amplified RNA was hybridized to Affymetrix Human Genome Focus arrays. Differentially expressed genes were identified using permutation t-test. MAIN OUTCOME MEASURE(S): Semen analyses and differential spermatozoal gene expression patterns measured by microarray. RESULT(S): Mean semen volume was not different between control and patients with cryptorchidism. Mean sperm density was significantly decreased between control, unilateral, and bilateral cryptorchid samples (110 vs. 87 vs. 16 million/mL). From the microarray expression data, we identified 43 genes differentially expressed between the two groups. Thirty-eight genes were significantly underexpressed in the cryptorchid samples including many transcriptional factors (cul3, prm1, hspcd35) and a testis-specific cell-adhesion gene (tpx-1) involved in germ cell maturation and sperm tail formation. An antiapoptotic gene (TNFAIP3) was highly overexpressed in the cryptorchid samples. CONCLUSION(S): Gene expression profiles offer insight into the diverse alterations that occur in cryptorchidism. The observed changes in spermatozoal expression of transcriptional and antiapoptotic genes may result in poor seminal parameters in formerly cryptorchid males.

Intracytoplasmic sperm injection (ICSI) with transmission of a ring(Y) chromosome and ovotesticular disorder of sex development in offspring.

We present a newborn infant with ovotesticular disorder of sex development and sex chromosome mosaicism with a supernumerary ring(Y), and a normal female cell line (47,XXr(Y)[10]/46,XX[40]. The ring (Y) was inherited from the child's father, and was transmitted following assisted reproductive technology and intracytoplasmic sperm injection (ICSI). The father presented with infertility and oligospermia, but cytogenetic analysis had not been carried out as part of the infertility workup. The Y containing cell line had not been seen on amniocentesis, which had shown a 46,XX apparently normal female karyotype in all cells studied. Molecular analysis using polymorphic probes from the X chromosome demonstrated that the 47,XXr(Y) cell line in the child was consistent with inheritance from the father, following meiosis I paternal non-disjunction. This report underscores the need to obtain chromosome analysis in couples with infertility who undergo assisted reproduction.

Partial bladder outlet obstruction selectively abolishes protein kinase C induced contraction of rabbit detrusor smooth muscle.

PURPOSE: Despite the acute onset, partial bladder outlet obstruction in the rabbit induces detrusor remodeling similar to that in men with benign prostatic hyperplasia in terms of its impact on structural and functional alterations in smooth muscle. We determined if partial bladder outlet obstruction induced remodeling alters the protein kinase C signaling pathway that leads to contraction. MATERIALS AND METHODS: Smooth muscle from control animals and those subjected to 2 weeks of partial bladder outlet obstruction were mounted for isometric force recording, measurement of myosin light chain phosphorylation and levels of adducin phosphorylation. Bladder muscle strips were stimulated by phorbol dibutyrate or carbachol in the presence and absence of bisindolylmaleimide-1. RESULTS: Smooth muscle strips from animals subjected to partial bladder outlet obstruction showed little to no increase in stress in response to phorbol dibutyrate and no increase in myosin light chain phosphorylation levels. Muscle strips from control animals produced a robust contraction with concomitant increases in myosin light chain phosphorylation. Inhibition of protein kinase C by bisindolylmaleimide-1 significantly depressed carbachol induced contractions of muscle strips from control animals but it had no effect on carbachol induced contractions of muscle strips from outlet obstructed animals. Phorbol dibutyrate increased phospho-adducin levels in muscle strips from the 2 animal sources, suggesting that protein kinase C could be activated. CONCLUSIONS: We propose that partial bladder outlet obstruction does not alter protein kinase C activation, but rather abolishes or uncouples the pathway(s) downstream of protein kinase C, leading to contraction. Loss of this pathway may contribute to the loss of normal voiding behavior and the resultant decompensated state.

Activation of the calcineurin pathway is associated with detrusor decompensation: a potential therapeutic target.

PURPOSE: We hypothesized that the calcineurin pathway mediated some of the complex remodeling process that allows a bladder subjected to partial outlet obstruction to adapt to its new workload. Atrial natriuretic factor mRNA expression served as a marker of calcineurin activation. MATERIALS AND METHODS: A total of 16 New Zealand White rabbits underwent surgical creation of partial outlet obstruction, followed by randomization to receive cyclosporin A (20 mg/kg intramuscularly twice daily) or no additional treatment for 14 days. Three animals underwent 2 weeks of partial bladder outlet obstruction followed by bladder biopsy and the reversal of obstruction. RESULTS: Atrial natriuretic factor expression was seen only in bladders with severe hypertrophy and it disappeared with the reversal of outlet obstruction. Cyclosporin A treatment resulted in a decrease in atrial natriuretic factor mRNA expression (p <0.05) and a marked shift in myosin heavy chain A-to-B ratios toward normal (p <0.01) and an increase in smooth muscle cross sectional area (p <0.05). Bladder mass decreased 40% but did not attain statistical significance (p = 0.08). CONCLUSIONS: The calcineurin pathway has a significant role in bladder wall hypertrophy following partial outlet obstruction. Bladder hypertrophy could not be fully prevented by cyclosporin A, suggesting that multiple signaling pathways are involved in this pathophysiology. The expression of myosin heavy chain AB isoforms is regulated in part by the calcineurin pathway.

Urethral strictures incident to bicycle motocross racing.

A dramatic shift from traditional team to alternative or "extreme" sports has given rise to a new generation of nontraditional athletes and sports-related injuries in the pediatric population. We present a case of 2 brothers who developed urethral strictures believed incident to BMX racing. We address current demographics and the general presentation and course of treatment to aid both the pediatric urologist and the general practitioner in prompt and proper diagnosis.

The physical characteristics of young males with varicocele.

OBJECTIVE: To determine if there is an association with habitus in young males with varicocele, as adolescent boys with varicoceles appear to be mostly taller and leaner than age-matched controls. PATIENTS AND METHODS: Retrospectively reviewing our records we obtained the height and weight of 43 consecutive males (mean age 14.3 years, range 11-19) under long-term follow-up for varicocele. The body mass index (BMI), heights and weights were compared with values from the respective growth charts for boys aged 2-20 years (Center for Disease Control and Prevention), and the statistical significance of differences determined using the chi-square test. RESULTS: The height and weight distributions of patients with varicocele indicated a significant deviation from normal in the 25-95th percentiles for stature and in the 25-75th for weight (P < 0.05). Deviations in BMI were insignificantly different from normal at each percentile. CONCLUSION: These results indicate that patients with varicocele are significantly taller and heavier than age-matched controls. Future studies to address the key areas identified in this study will help to further assess the distribution of the incidence of varicocele in closely defined subsets of adolescent growth and development, which may provide some insight into the cause of varicoceles.