Validation-verification of a highly effective, practical human testicular tissue in vitro culture-cryopreservation procedure aimed to optimize pre-freeze and post-thaw motility.

PURPOSE: The aim of our paper was to validate a testicular biopsy procedure that simplifies handling, processing, and cryopreservation, while at the same time optimizes sperm motility before freezing and after thawing. METHODS: Two prospective studies were conducted to verify, optimize, and understand the virtues of pre-freeze testicular tissue IVC at different temperatures (21, 30, or 37 °C). Testicular tissue was obtained from clinical specimens designated for whole tissue cryopreservation (i.e., intact mass of tubules) and/or for fresh use in IVF-ICSI cycles. Whole testicular biopsy pieces (1-3 mm(3)) were diluted in glycerol containing freeze solutions, slow cooled to 4 °C and then rapidly frozen in LN2 vapor. Fresh and post-thaw testicular biopsy tissue were evaluated for changes in the quantity (%) and pattern of motility (I-IV: twitching to rapid progression, respectively) over a 1 week duration. The clinical effectiveness of IVC-cryopreserved whole testicular biopsy tissue was also validated analyzing fresh embryo transfers. RESULTS: More reliable recovery of motile testicular sperm was achieved using whole tissue freeze preservation combined with IVC (24-96 h) post-acquisition at an incubation temperature of 30 °C compared to ambient temperature (21 °C) or 37 °C. Up to 85 % of the pre-freeze motility was conserved post-thaw (+3 h) for easy ICSI selection. Sperm longevity was optimized to fresh tissue levels by implementing testicular biopsy sucrose dilution post-thaw. Favorable clinical outcomes were proven using frozen-thawed testicular biopsy sperm for ICSI. CONCLUSIONS: By employing minimal tissue manipulation, integrating pre-freeze IVC processing at 30 °C and the freezing of whole testicular biopsy tissue, we have reduced the labor and improved the efficacy of processing testicular tissue for freeze-preservation and subsequent ICSI use.

Circumflex artery ventricular fistula and pseudoaneurysm after mitral reoperation.

Prosthetic mitral valve reoperation complicated by atrioventricular groove pseudoaneurysm and circumflex ventricular fistula is presented. Ligation of the circumflex artery during mitral valve replacement is implicated after review of a previous cardiac angiogram.

Adenoviral-p53 gene transfer to orthotopic and peritoneal murine bladder cancer.

PURPOSE: This study was designed to examine the potential for adenoviral-mediated gene therapy in primary and metastatic bladder cancer. MATERIALS AND METHODS: Orthotopic and intraperitoneal bladder tumors were established after delivery of 1 x 10(6) MBT-2 cells into syngeneic mice. Gene transfer was accomplished via intravesical or intraperitoneal instillation by using an E-1 deleted adenovirus encoding LacZ or human p53. Successful tumor transduction was confirmed in tumor DNA and mRNA by polymerase chain reaction. Detection of recombinant gene product was detected by histochemical staining (X-gal) and Western blot. RESULTS: Palpable tumors developed 18 days following implantation. LacZ and p53 mRNA were present in tumor and adjacent normal tissue after bladder and intraperitoneal vector administration. Recombinant gene products were identified by histochemistry and Western blot. CONCLUSION: Bladder tumor-directed gene transfer using adenoviral vectors is an efficient and powerful tool for evaluating the adjuvant role of therapeutic gene products.

Use of a prefabricated tunica vaginalis fascia flap to reconstruct the tunica albuginea after recurrent penile prosthesis extrusion.

PURPOSE: Although a penile prosthesis usually perforates into the urethra, it can extrude through the glans or corporeal shaft. Various materials have been used to reconstruct tunica albuginea but no method of repair has been satisfactory in such difficult cases. Repair of the weakened tunica albuginea should ideally be performed with autogenous tissues. Inasmuch as the scarred tissue bed is inadequate to ensure graft survival and no local flaps are available for this purpose, prefabrication of a local flap has been designed. MATERIALS AND METHODS: We present 2 cases in which the distal corpus was reconstructed with a prefabricated tunica vaginalis fascia flap. The first stage involves grafting rectus fascia onto the external tunica vaginalis of the testicle. At the second stage the prefabricated tunica vaginalis fascia flap is transposed to the distal corpus, placing it as a buttress between the cylinder and urethra medially or between the cylinder and thin lateral and distal tunica albuginea. The flap also replaces part of the tunica albuginea. RESULTS: In both patients repair of the tunica albuginea was successful and each has a functioning inflatable penile prosthesis at 2 1/2 1 1/2 years postoperatively, respectively. CONCLUSIONS: Reconstruction of the weak tunica albuginea with a prefabricated tunica vaginalis fascia flap is an excellent procedure in these difficult cases.

Adenoviral-mediated gene transfer to bladder in vivo.

This study was designed to examine the potential for gene therapy in bladder in vivo using adenoviral vectors. Gene transfer to rat bladders was accomplished via direct intravesical instillation using a replication-defective adenoviral vector containing a marker gene encoding for Escherichia coli beta-galactosidase (beta-gal). Successful gene transfer was confirmed by analyzing bladder samples for DNA and RNA using polymerase chain reaction (PCR) with primers specific for beta-gal and adeno sequences, detecting beta-gal in full-thickness bladder wall using specific histochemical staining (X-gal) and documenting recombinant protein production. Bladder architecture was preserved, without evidence of distant spread of virus as assessed by PCR. Gene expression was evident for at least 7 days. In summary, bladder cells can be genetically altered using replication-deficient adenoviral vectors via simple intravesical instillation of vector. Introduction of exogenous genetic material is a potentially powerful therapeutic modality for immunomodulation of bladder neoplasms.