L. Ralph Rohr

Commonly occurring loss and mutation of the MXI1 gene in prostate cancer.

One of the most common chromosomal abnormalities in prostate cancer involves loss of 10q22–qter. Rarely, a smaller deletion, involving 10q24–q25, has been observed, suggesting the presence of a tumor suppressor gene at this site. We previously demonstrated that the MXI1 gene maps to 10q24–q25 and is mutated in some tumors with cytogenetically detectable deletions of this locus. MXI1 encodes a basic‐helix–loop–helix protein that suppresses the transcriptional activity of the MYC oncoprotein by competing for the common dimerization partner, MAX, and binding to identical DNA sites. Because more than 90% of prostate tumors contain no cytogenetic abnormality of 10q, the relevance of MXI1 loss and/or mutation to the vast majority of cases remains unclear. We prospectively evaluated prostate tumors for loss of MXI1 by fluorescence in situ hybridization (FISH) and cytogenetic techniques. Twenty‐one of 40 tumors (53%) demonstrated loss of a single MXI1 allele as determined by FISH. Ten cases with cytogenetically normal 10qs, but with FISH‐documented deletion of MXI1, were examined at the molecular level, and eight mutations were identified, albeit at low frequency. Five of the mutant proteins were unable to bind DNA in association with MAX. We conclude that MXI1 gene loss in prostate cancer is common and most frequently involves a cytogenetically undetectable deletion. Genes Chromosomes Cancer 22:295–304, 1998.

Renal cryoablation in a canine model

OBJECTIVES To assess the potential safety and utility of cryoablation for treatment of selected renal tumors in a canine model. METHODS Ultrasound and direct physical measurements (depth and width) of five cryolesions were compared. Cryolesions were examined histologically in 6 animals, which were killed at 4 hours, 2 days, 1 week, 3 weeks, 6 weeks, and 12 weeks. Mortality/morbidity was assessed in 12 animals over a 1-month interval, where 6 animals received small (approximately 2 cm) cryolesions and 6 animals received large (one third to one half of kidney) cryolesions. Laparoscopic cryoablation was performed in 2 animals. RESULTS A statistically significant association of physical and ultrasound dimensions was observed (correlation coefficient R = 0.9295; P = 0.0001). Histologic studies in animals killed up to 1 week after cryoablation revealed complete coagulative necrosis within the cryolesion. The boundary transition from normal to complete tissue necrosis occurred in 1 to 2 mm. Animals killed 3 weeks to 3 months after cryoablation revealed progressive organization with granulation tissue, chronic inflammation, hemosiderosis, fibrosis, and contraction of the cryolesion with parenchymal loss. Untreated renal tissue was histologically normal in all kidneys. No mortality or morbidity was detected in the 12 animals followed for 30 days regardless of the size of the cryolesion. Laparoscopic cryoablation was performed successfully in 2 animals without modification of standard laparoscopic methods. CONCLUSIONS Sonographic, histologic, and laparoscopic data in a canine model suggest that cryoablation may be a safe, feasible, and useful method for treatment of selected renal neoplasms.

A Comparison of Immunohistochemical Stain Quality in Conventional and Rapid Microwave Processed Tissues

Same-day turnaround of pathology specimens is desirable in this era of managed care, and rapid microwave tissue processing produces histologic features of a quality equivalent to overnight processing. We studied whether microwave-assisted rapid tissue processing adversely affects the quality of immunohistochemical staining. We selected 30 specimens (20 neoplastic and 10 nonneoplastic) from our routine surgical pathology workload. Paired large tissue blocks were made from each specimen type, one for microwave-assisted rapid processing and one for conventional processing. Two microarrays of 60 punches each were made from the donor blocks. The microarray blocks were examined for intensity and extent of staining by 44 commonly used antibodies. Slides were reviewed independently by 2 pathologists blinded to the type of processing used. In 5,280 tissue punches examined, we found a high degree of concordance in quality, as measured by intensity and extent of immunohistochemical staining, between microwave and routinely processed tissues. Our study demonstrates that quality of immunohistochemical staining is similar between rapid microwave and conventional processing. The potential need for immunohistochemical analysis is not a contraindication for microwave-assisted rapid tissue processing.

Chromosome 7 Abnormalities in Prostate Cancer Detected by Dual-Color Fluorescence In Situ Hybridization

Aneusomy of chromosome 7 and loss at 7q (especially 7q31.1) have been reported in prostate cancer. To further investigate abnormalities of 7q and the relationship with whole chromosome 7 changes, we have conducted a dual-color fluorescence in situ hybridization (FISH) analysis on isolated nuclei from 28 primary prostate cancers. A pericentromeric probe for chromosome 7, five newly isolated sequence-specific bacterial artificial chromosome (BAC) probes from 7q31.1, and one BAC for the epidermal growth factor receptor (EGFR) gene at 7p12 were used in dual color hybridizations. Pericentromeric probes for chromosomes X and 4 were also used as controls. Sixteen (57.1%) of the 28 tumors showed clonal aberrations. Nine of them were trisomy 7 and four were hypertetrasomy for chromosome 7. Deletions at 7q31.1 were found in two of the high grade tumors. With the exception of these two cases, all other cases showed concordant results using all probes. These findings confirm previous studies that aneusomy of 7 is associated with prostate cancer progression, and there may be a tumor suppressor gene (TSG) at 7q31.1 which is associated with tumor progression. In addition, our study indicates: (1) the deletion pattern of individual nuclei infers that deletions at 7q31.1 precede reduplications of chromosome 7; and (2) the amplification of EGFR was not detected at the DNA level, suggesting that activation of this oncogene may play a minor role in prostate cancer.

Microdissection, DOP-PCR, and comparative genomic hybridization of paraffin-embedded familial prostate cancers.

There is a clear genetic component to prostate cancer susceptibility. Regions reported to be linked to prostate cancer include 1q24-25 (HPC-1), 1q42.2-43, and Xq27-28. There is limited genetic information on familial prostate tumors. We used the Utah Population Database to identify familial prostate cancer cases and selected 35 cases from high-risk families. Tissue blocks containing discernable tumor were available from 19 cases; 13 of these yielded adequate specimens for analysis. Six cases came from families with linkage to HPC-1, 3 were known to have linkage to Xq27-28, and 4 had no linkage to a known locus; 7 cases were analyzed from patients who showed no known linkage (sporadic tumors) as controls. These paraffin-embedded tumors were laser microdissected, degenerate oligonucleotide (DOP)-amplified, and labeled for fluorescence detection by comparative genomic hybridization (CGH). Loss of 7q, 10q, and 16q and gain of 8q were common abnormalities present in both familial and sporadic tumors. Distinctive abnormalities included loss of 3p12-3p22 in 3 of 6 HPC-7-linked cases and in 2 of 3 X-linked cases and gain of 6q11-6q21 in 2 each of HPC-1 and X-linked tumors. In conclusion, laser microdissection, DOP-PCR, and CGH is a feasible method for analysis of paraffin-embedded prostate tumors. This study provides preliminary data suggesting that familial prostate cancer harbors some unique genetic changes when compared with sporadic prostate tumors.

Resorption Rate and Biocompatibility Characteristics of Two Polyester Ventilation Tubes in a Guinea Pig Model

OBJECTIVES: Determine the resorption rate and biocompatibility characteristics of 2 polyester ventilation tubes, and to determine whether soap and water exposure accelerates polyester tube degradation. STUDY DESIGN AND SETTING: 50/50 poly (D, L-lactide-co-glycolide; PLGA-50) and poly (L-lactide; PLA) polymers were placed into the tympanic membranes of Hartley pigmented guinea pigs. Integrity of the tubes was determined by weekly otoscopic examination. Biocompatibility was assessed by comparing auditory brainstem response (ABR) thresholds and by examining tympanic membrane changes following tube resorption. Shah minigrommet ventilation tubes were used as controls. In the second portion of this study, implanted PLGA-50 and PLA tubes were exposed weekly to a mixture of soap and water (1:5) until complete resorption was observed. Biocompatibility was assessed by periodic ABR testing and tympanic membrane examination. RESULTS: The PLA tubes remained in the tympanic membrane for a longer period (63.2 ± 19.3 days) than the PLGA-50 (18.8 ± 8.1 days). The tympanic membrane and resorbable tube interface demonstrated equivalent findings for auditory thresholds and tissue histopathology at the implant site compared to nonresorbable controls. The resorption behavior was not altered by exposure to soap and water. Tympanic membranes of all animals following tube degradation and soap water exposure were intact with minimal scarring and no signs of persistent foreign body response. The histological analysis showed that implantation of resorbable tubes was not accompanied by secondary infection with otorrhea through the tube, did not result in a permanent perforation or dislocation of the tube into the middle ear cavity, and was not followed by excess tympanosclerosis or localized or diffuse membrane atrophy. CONCLUSIONS AND SIGNIFICANCE: Resorbable polyester pressure equalization tubes demonstrate predictable resorption behavior and similar bio-compatibility characteristics when compared with nonresorbable Shah minigrommet ventilation tubes. Exposure to soap water does not accelerate polyester tube degradation nor change the host tissue response during the indwelling period or after complete resorption. The data suggests that resorbable ventilation tubes are substantially equivalent to other FDA-approved tympanostomy devices with regard to safety and biocompatibility in the guinea pig model examined and may provide improved clinical performance by combining this approach with sustained release technology. EBM rating: B-2.