Vytautas Bilius

Intravesical cidofovir to treat BK virus‐associated hemorrhagic cystitis in children after hematopoietic stem cell transplantation

HC related to BK virus replication might be a severe complication following allogeneic HSCT. There are no clearly defined treatment guidelines in pediatric population. The data on the effectiveness of ICI to manage severe bleeding in children are very limited. We report our experience of intravesical cidofovir in four children, 6–15 yr of age, to manage grade III–IV BK virus‐associated HC. Three of four children had high CSA serum level prior to developing cystitis. Intravesical instillations of cidofovir resulted only in temporal relief of bleeding. After immune suppression was withdrawn or tapered, intravesical instillations of formalin solution had to be undertaken to abort severe bleeding. We concluded that intravesical cidofovir alone did not appear to be sufficiently effective in case of severe HC, necessitating complimentary procedures to stop macrohematuria.

Gene Expression Changes Underlying Idiopathic Central Hypogonadism in Cryptorchidism with Defective Mini-Puberty

The whole genome RNA profiling of testicular biopsies by DNA strand-specific RNA sequencing was examined to determine a potential causative role of isolated congenital cryptorchidism in azoospermia and/or infertility in the context of our previously published GeneChip data. Cryptorchid patients, aged 7 months to 5 years and otherwise healthy, were enrolled in this prospective study. During surgery, testicular tissue biopsies were obtained for histological examination and RNA sequencing. Fifteen patients were selected based on the histological results and were divided into 2 groups. Seven were classified as belonging to the high infertility risk (HIR) and 8 to the low infertility risk (LIR) group. Cryptorchid boys in the HIR group lacked transformation of gonocytes into Ad spermatogonia due to impaired mini-puberty. This group of patients will be infertile despite successful surgery. The new important finding was a decreased PROK2, CHD7, FGFR1, and SPRY4 gene expression in the HIR group. Furthermore, identification of multiple differences in gene expression between HIR and LIR groups underscores the importance of an intact hypothalamic-pituitary-gonadal axis for fertility development. Our RNA profiling data strongly support the theory that in the HIR group of cryptorchid boys insufficient PROK2/CHD7/FGFR1/SPRY4 gene expression induces deficient LH secretion, resulting in impaired mini-puberty and infertility. We therefore recommend hormonal treatment for this cohort of cryptorchid boys with defective mini-puberty following a seemingly successful orchidopexy.

GnRHa Treatment of Cryptorchid Boys Affects Genes Involved in Hormonal Control of the HPG Axis and Fertility

The gonadotropin-releasing hormone agonist (GnRHa; Buserelin) rescues fertility during adulthood in the majority of high infertility risk cryptorchid boys presenting with defective mini-puberty. However, the molecular events governing this effect are not understood. We report the outcome of an RNA profiling analysis of testicular biopsies from 4 operated patients who were treated with GnRHa for 6 months versus 3 operated controls who were not treated. GnRHa induces a significant transcriptional response, including protein-coding genes involved in pituitary development, the hypothalamic-pituitary-gonadal axis, and testosterone synthesis. Furthermore, we observed an increased abundance of long noncoding RNAs (lncRNAs) participating in epigenetic processes, including AIRN, FENDRR, XIST, and HOTAIR. These data are consistent with the hypothesis that hypogonadotropic hypogonadism in boys with altered mini-puberty is the consequence of a profoundly altered gene expression program involving protein-coding genes and lncRNAs. Our results point to molecular mechanisms that underlie the ability of GnRHa to rescue fertility.

Incidence of High Infertility Risk among Unilateral Cryptorchid Boys

Background: Increasing evidence of progressive damage to germ cell development in boys with cryptorchidism suggests recommending surgery until one year of age. However, despite early and successful orchidopexy, cryptorchid boys with impaired mini-puberty will suffer from infertility. We reviewed changes in the timing of surgery during the past decade and the incidence of unilateral cryptorchid boys with defective mini-puberty. Methods: Medical registries were reviewed for all patients who were operated on for cryptorchidism at the main pediatric urological center of the country. The ages of surgery in cases of unilateral cryptorchidism were compared between the years 2000-2001 and 2012-2013. A high risk of infertility was considered when no Ad spermatogonia were found. Two groups were compared: group I - operated on until the age of 1.5 years and group II - older than 1.5 years. Results: The average age at operation decreased from 5.3 to 4.1 years. Forty-six biopsies in boys with unilateral cryptorchidism were made during orchidopexy on undescended testicles. Overall, 44% in group I and 50 % in group II (p > 0.05) had no Ad spermatogonia. Conclusions: The average age of operation for cryptorchidism has decreased, but remains far above the recommended age. The high prevalence of histologically proven risk of infertility underscores the necessity of more education regarding the importance of earlier surgery and the research on hormonal prevention of infertility.

DMRTC2, PAX7, BRACHYURY/T and TERT Are Implicated in Male Germ Cell Development Following Curative Hormone Treatment for Cryptorchidism-Induced Infertility

Defective mini-puberty results in insufficient testosterone secretion that impairs the differentiation of gonocytes into dark-type (Ad) spermatogonia. The differentiation of gonocytes into Ad spermatogonia can be induced by administration of the gonadotropin-releasing hormone agonist, GnRHa (Buserelin, INN)). Nothing is known about the mechanism that underlies successful GnRHa treatment in the germ cells. Using RNA-sequencing of testicular biopsies, we recently examined RNA profiles of testes with and without GnRHa treatment. Here, we focused on the expression patterns of known gene markers for gonocytes and spermatogonia, and found that DMRTC2, PAX7, BRACHYURY/T, and TERT were associated with defective mini-puberty and were responsive to GnRHa. These results indicate novel testosterone-dependent genes and provide valuable insight into the transcriptional response to both defective mini-puberty and curative GnRHa treatment, which prevents infertility in man with one or both undescended (cryptorchid) testes.

Genes Involved in Long-Term Memory Are Expressed in Testis of Cryptorchid Boys and Respond to GnRHa Treatment

It has been known for many years that boys with unilateral or bilateral undescended testis (cryptorchidism) tend to have a low IQ, and those who belong to the high infertility risk (HIR) group perform less well at school than low infertility risk (LIR) patients. However, the molecular biological processes underlying this phenomenon are not understood. In this study, we report the outcome of testicular RNA profiling for genes involved in long-term memory formation. We analyzed the histology and the transcriptome of testicular biopsies from bilateral HIR cryptorchid boys, comparing those who received GnRHa treatment for 6 months after the first surgery with those who did not receive GnRHa before the second surgery. We found that GnRHa treatment alters the testicular mRNA levels of neuronal genes that are involved in long-term memory and testosterone synthesis. These data highlight a possible molecular link between cryptorchidism, impaired mini-puberty, and diminished cognitive functions. Our results are consistent with the hypothesis that hypogonadotropic hypogonadism in cryptorchid boys with altered mini-puberty may affect neuronal genes important for memory and learning, which could help explaining the negative correlation between cryptorchidism and intellectual abilities.

Infertility indicators in the biopsies from undescended testicles in boys of different ages

Objectives. To compare the risk of infertility in boys operated for cryptorchidism at different ages. Patients and methods. Thirty patients with unilateral undescended testis underwent orchidopexy and were subjected to ipsilateral testicular biopsy from 2010 to 2012 in department of Pediatric Urology, Children‘s Hospital, Affiliate of Vilnius University Hospital Santariskiu klinikos. Biopsies were fixed in 2% glutaraldehyde, stained with toluidine blue, sectioned and analyzed by lightmicroscopy at 400× magnification. The number of Ad spermatogonia was assessed. Patients were classified into 2 groups according to their age: I–<2 years of age, II – ≥2 years of age. There were 14 boys in group I and 16 boys in group II. Median age in group I was 1.47 year, and 6.91 years in group II. Results. Adult dark spermatogonia was found in the biopsies of 14 (46.6%) patients with a count 0.09-0.86 Adult dark spermatogonia per tubule. Adult dark spermatogonia were absent in 16 (53.3%) biopsies of cryptorchid boys: 5 in group I, and 11 in group II. Patients with no Ad spermatogonia had a lower tubular density. Conclusions. The best results of testicular histology were in group 1. These boys had lower risk of infertility . Data of our biopsies support the idea of progressive domage to germ cell development with increasing age. This is alarming and emphasizes the need of improvement for the treatment of cryptorchidism and children examinations.

Analysis of Trends in Hypospadias Treatment from 1991 to 2010

Objective: to compare the choice of operative technique and the rate of reoperations after the first hypospadias repair at our department during the last two decades. Materials and methods: medical records of all patients operated for hypospadias during the period of 1991 – 2010 were reviewed. We included patients with distal, penile and proximal hypospadias. The choice of operating technique, the amount of reoperations and the age at first operation were compared for last two decades (1991 – 2000 and 2001 - 2010). Results: a total of 965 patients met the inclusion criteria. 60% of patients had no reoperations, 27,7% had one and 12,3% had two or more reoperations during the period of 1991 - 2000. 73% had no reoperations, 20,4% had one and 6,5% had two or more reoperations during the period of 2001 - 2010. We analyzed the mean age at first operation: during the first decade it was 5,4 years and 4,1 – during the second. Conclusions: Complication rate after hypospadias repair at our department is decreasing but remains relatively high. Long term follow-up and the analyzis of results are necessary in order to improve our results.

Early Surgical Complications after Renal Transplantation in Children

Objective. To assess early surgical complications after renal transplantation in children treated in Pediatric Department of Children’s Hospital, the Affiliate of Vilnius University Hospital Santariskiu Klinikos after renal transplantation and determine their time of origin, symptoms and to compare the result with global literature data. Materials and methods. We retrospectively analyzed 38 children’s after 42 kidney transplantation case histories and searched for early surgical complications during first 3 month after renal transplantation. We searched for kidney vascular thrombosis, urine leak, ureteral stenosis, lymphocele, wound infection and renal artery stenosis. Patients’ age, time, when complication developed , signs, treatment and outcome results were assessed. Results. Kidney vascular thrombosis was found in 3 cases (7.3%), hydronephrosis due to ureteral stenosis in 4 cases (9.8%), 3 of them treated conservative, and 1 – by internal stent, wound infection in 2 cases (4.9%), and renal artery stenosis in 1 case (2.4%). In all cases after renal vascular trombosis the transplant was lost. Conclusion. The frequency of surgical complications after renal transplantation in children is less than refers medical literature, but this may be due to small number of transplantations. Nevertheless, surgical complications remain a major cause of graft loss in children‘s kidney transplantation in the early period after transplantation. Article in Lithuanian