DISSEMINATED INVASIVE ASPERGILLOSIS IN A PATIENT WITH ADRENOCORTICAL CARCINOMA.

OBJECTIVE: To describe the course and anatomo-pathologic findings in a patient with a cortisol-producing adrenocortical carcinoma (ACC) who presented with disseminated invasive aspergillosis (DIA). We also review the clinical characteristics of ACC and the pathogenesis of DIA in the setting of hypercortisolemia. METHODS: We describe the case of a 47-year-old man with a recent diagnosis of ACC associated with hypercortisolism, who presented with a large right basal ganglia hematoma and lesions on multiple organs thought to be metastatic. The patient deteriorated rapidly and unfortunately 6 days after presentation he died. An autopsy was performed which revealed DIA as the cause of death. RESULTS: DIA in ACC was reported for the first time in 1981. Since then, no other case has been reported in the literature associating these 2 conditions. The lesions this patient presented on images were difficult to differentiate from metastatic lesions. The lack of inflammatory features due to hypercortisolemia accounted for this. Both ACC and DIA have very high mortality rates, even when DIA is treated appropriately. CONCLUSION: Opportunistic fungal infections are common in hypercortisolemia, however disseminated and invasive forms are rare. Awareness of uncommon presentations of hypercortisolemia is important for early treatment of those patients with a chance of survival.

Renal subcapular saline injection during percutanoues renal cryoablation to prevent bowel injury.

INTRODUCTION: To evaluate the feasibility of percutaneous injection of saline in the renal subcapsular space to prevent bowel injury and histopathologic effects of bowel cryolesion during renal cryoablation in a porcine model. MATERIAL AND METHOD: Six pigs underwent percutaneous renal cryoablation with two freeze cycles in the lower pole of both kidneys. Six kidneys were injected with 10 ml saline into the renal subcapular space before cryoablation. The bowel was brought into contact with the edge of the ice ball with laparoscopic assistance during renal cryoablation, on the side with saline injection as well as on the control side. One of these animals was kept for survival follow-up and laparotomy for 7 days post cryoablation. The bowel cryolesion sites were observed and compared based on the presence or absence of renal subcapsular saline injection. RESULTS: The mean diameter of acute bowel injury with and without saline renal subcapsular injection was 7.25 +/- 1.26 and 14.5 +/- 0.58 mm, respectively. The influence of injecting a saline buffer was a significant decrease in the bowel cryolesion compared to controls (p = 0.0003). In addition, a pig kept for follow-up confirmed no bowel perforation after 7 days at a site that was cryolesioned on the side with renal subcapsular saline injection, but sustained bowel perforation in another segment lesioned by contact with a kidney without a saline injection. Gross and microscopic pathological examination was consistent with these interpretations. CONCLUSION: Preliminary results in a porcine model show that percutaneous renal subcapsular saline injection is a feasible and promising technique for preventing bowel complications of percutaneous image-guided renal cryoablation.

Thermal maps around two adjacent cryoprobes creating overlapping ablations in porcine liver, lung, and kidney.

PURPOSE: To determine cryoprobe spacing requirements in order to achieve overlapping ablation zones using the same ablation protocol in porcine liver, lung, and kidney. MATERIALS AND METHODS: Six female pigs underwent cryoablation of the liver, lung, and kidney. Two 2.4-mm cryoprobes were spaced 20-mm apart with seven 16-gauge thermometers placed linearly and in axis with the cryoprobes at 5-mm increments from one another. The placement of the thermometers was such that three were placed between the two probes and two were placed laterally to each probe. Simultaneous use of the cryoprobes, using 12- and 8-minute double-freeze cycles, was performed with intratissue temperature monitoring during the procedure. RESULTS: The center temperatures between the two cryoprobes in kidney, lung, and liver were -25.87( degrees )C +/- 1.91, -6.47(degrees )C +/- 3.94, and 0.48( degrees )C +/- 6.69, respectively. Dual 2.4-mm cryoprobes in our model achieved acute pathological complete coagulative necrosis zone at the center of the ablation zone between the cryoprobes only in the kidney tissue where a mean diameter of the acute complete coagulative necrosis zone was 39.6 mm +/- 0.76 mm. CONCLUSIONS: The critical temperature of -20 degrees C was not reached at the midpoint between the probes with the 20-mm spacing arrangement in the lung and liver. These results emphasize the need for individualized organ ablation treatment protocols.

Differences in ablation size in porcine kidney, liver, and lung after cryoablation using the same ablation protocol.

OBJECTIVE: The purpose of our study was to assess the variation in size of acute necrosis and the variation in thermal map measured during cryoablation in multiple organs using the same ablation protocol for each organ. MATERIAL AND METHODS: Eight female pigs underwent one cryoablation per organ of kidney, lung, and liver performed with open surgery with a 2.4-mm cryoprobe. A 12- and 8-minute double-freeze cycle was used. Intratissue temperatures were monitored using 16-gauge thermometers spaced at 5.0-mm increments from the cryoprobe. The comparison of results among tissues was performed using the multiple analysis of variance. The -20 degrees C thermal diameter was correlated with tissue damage. The kidneys, lungs, and liver were removed and examined histologically for a pathologic complete coagulative necrosis zone. RESULT: A single 2.4-mm cryoprobe had a mean ice ball diameter in kidney, lung, and liver of 38.5 +/- 4.7, 35.5 +/- 3.6, and 32.5 +/- 2.7 mm, respectively. A mean -20 degrees C thermal diameter was achieved at 24.07 +/- 1.38 mm in kidney, 12.76 +/- 3.0 mm in lung, and 8.8 +/- 3.7 mm in liver by means of regression analysis. The acute pathologic complete coagulative necrosis zone size was 21.0 +/- 1.56 mm (kidney), 11.6 +/- 1.48 mm (lung), and 8.0 +/- 1.20 mm (liver). CONCLUSION: The inherent characteristics of different organs manifest different ablation zone sizes during cryoablation despite the same ablation protocol being used. This information should be factored into planning for ablation procedures.

Detection of human papillomavirus-16 in fine-needle aspirates to determine tumor origin in patients with metastatic squamous cell carcinoma of the head and neck.

PURPOSE: Patients with head and neck squamous cell carcinoma (HNSCC) often clinically present with metastases to regional lymph nodes. Fine-needle aspiration of neck masses is routinely used to establish the presence of metastatic carcinoma and in turn to initiate a subsequent workup to determine the site of tumor origin. Human papillomavirus (HPV) 16 is an important etiologic agent for HNSCCs that arise from the oropharynx but less so for tumors from non-oropharyngeal sites. HPV16 detection thus provides a strategy for localizing an important subset of HNSCCs, but this approach has not been applied to fine-needle aspiration specimens. EXPERIMENTAL DESIGN: We did in situ hybridization for HPV16 on 77 consecutive aspirated neck masses diagnosed as metastatic squamous cell carcinoma. P16 immunohistochemistry was also done because p16 overexpression may serve as a surrogate marker of HPV-associated HNSCC. RESULTS: HPV16 was detected in 13 of the 77 (17%) aspirates. By site of origin, HPV16 was detected in 10 of 19 metastases from the oropharynx but in none of 46 metastases from other sites (53% versus 0%; P < 0.0001). HPV16 was not detected in 2 branchial cleft cysts misdiagnosed as metastatic squamous cell carcinoma, but it was detected in 3 of 10 metastases from occult primary tumors. P16 expression was associated with the presence of HPV16: 12 of 13 HPV16-positive metastases exhibited p16 expression, whereas only 4 of 62 HPV16-negative metastases were p16 positive (92% versus 6%; P < 0.0001). P16 expression also correlated with site of tumor origin: 13 of 19 oropharyngeal metastases were p16 positive, whereas only 1 of 46 non-oropharyngeal metastases was p16 positive (68% versus 2%; P < 0.0001). CONCLUSIONS: HPV16 status can be determined in tumor cells aspirated from the necks of patients with metastatic HNSCC. Its presence is a reliable indicator of origin from the oropharynx.

Prophylactic gelatin sponge tract injection to prevent bleeding after percutaneous renal cryoablation in a swine model.

PURPOSE: Achievement of hemostasis can be a challenge during percutaneous renal cryoablation (PRC). This study describes the use of a porcine model to test the ability of gelatin sponge injection into the tract to limit bleeding at the site of cryoprobe puncture. MATERIALS AND METHODS: A total of nine swine (18 kidneys) underwent bilateral ultrasound-guided PRC with double freeze/thaw cycle protocol with use of a 2.4-mm cryoprobe. The cryoablation location and protocol were applied identically to both kidneys in each pig; however, only one side received gelatin sponge injection after cryoablation through a coaxial sheath (3 mm). After removal of the sheath, a midline laparotomy incision was performed and sponges were placed around the kidneys. Blood loss was measured by calculating the change in weight of the sponges after 30 minutes of absorption time. Acute blood loss was compared between the two groups. The kidneys treated with gelatin sponge were removed and grossly and histologically examined to identify the gelatin sponge at the puncture cryolesion. RESULTS: The use of gelatin sponge resulted in significantly less blood loss (mean, 8.24 mg +/- 4.68) compared with the control kidneys (mean, 20.24 mg +/- 8.14; P = .001). Gross and histopathologic results confirmed that the gelatin sponge was in the cryoablation puncture sites. Mean diameters of cryoablation ice balls on the gelatin sponge and control sides were 3.9 cm +/- 0.2 and 3.8 cm +/- 0.4, respectively. The lesions were not significantly different between sides. CONCLUSION: Percutaneous tract injection of gelatin sponge appears promising as a method to decrease acute blood loss from PRC.

Percutaneous bladder cryoablation in porcine model.

PURPOSE: To demonstrate the morphologic changes of full-thickness bladder cryoablation utilizing contemporary percutaneous technology. MATERIALS AND METHODS: Cryoablation of the bladder wall was conducted in 24 pigs. The bladders were exposed laparoscopically and inspected cystoscopically. The animals underwent either extravesical (serosal) or transvesical (mucosal) approaches for the creation of the cryoablation lesion. Single or double freeze/thaw cycles were applied, and no bladder drainage was used. The bladder was evaluated for perforation, and histologic examination was undertaken to assess the extent of acute, subacute (2 weeks), or chronic (1 month) lesions. RESULTS: Cryoablation reliably produced a controlled transmural area of necrosis (both serosa and mucosa) by both techniques. The size and extent of the lesion were directly proportional to the duration of freezing and the type of cryoprobe used. The pathology report confirmed the full-thickness coagulative necrosis of muscle. Bladder perforation was not detected clinically or at autopsy. CONCLUSION: Cryoablation produces reliable zones of tissue destruction without bladder perforation. These preliminary data show the safety and feasibility for trials in the treatment of bladder tumors by laparoscopic, cystoscopic, or image-guided techniques.

A molecular classification of papillary renal cell carcinoma.

Despite the moderate incidence of papillary renal cell carcinoma (PRCC), there is a disproportionately limited understanding of its underlying genetic programs. There is no effective therapy for metastatic PRCC, and patients are often excluded from kidney cancer trials. A morphologic classification of PRCC into type 1 and 2 tumors has been recently proposed, but its biological relevance remains uncertain. We studied the gene expression profiles of 34 cases of PRCC using Affymetrix HGU133 Plus 2.0 arrays (54,675 probe sets) using both unsupervised and supervised analyses. Comparative genomic microarray analysis was used to infer cytogenetic aberrations, and pathways were ranked with a curated database. Expression of selected genes was validated by immunohistochemistry in 34 samples with 15 independent tumors. We identified two highly distinct molecular PRCC subclasses with morphologic correlation. The first class, with excellent survival, corresponded to three histologic subtypes: type 1, low-grade type 2, and mixed type 1/low-grade type 2 tumors. The second class, with poor survival, corresponded to high-grade type 2 tumors (n = 11). Dysregulation of G1-S and G2-M checkpoint genes were found in class 1 and 2 tumors, respectively, alongside characteristic chromosomal aberrations. We identified a seven-transcript predictor that classified samples on cross-validation with 97% accuracy. Immunohistochemistry confirmed high expression of cytokeratin 7 in class 1 tumors and of topoisomerase IIalpha in class 2 tumors. We report two molecular subclasses of PRCC, which are biologically and clinically distinct and may be readily distinguished in a clinical setting.

Metastatic breast carcinoma involving the thyroid gland diagnosed by fine-needle aspiration: a case report.

Secondary involvement of the thyroid gland from a remote primary malignancy is uncommon. The distinction of metastatic carcinoma (MC) or sarcoma from a primary thyroid malignancy is important because the treatment is different. We discuss a case of a 64-yr-old female with a history of breast carcinoma, who presented with pain and swelling in her neck 5 yrs after being diagnosed with breast cancer. She had undergone mastectomy with subsequent chemotherapy and radiation for infiltrating mammary carcinoma. During the 5-yr interval, she had been free of clinically evident metastatic disease. Subsequent work-up revealed two distinct nodules in the left lobe of her thyroid gland as well as a subcutaneous mass in her right shoulder. A fine-needle aspiration (FNA) of the larger thyroid nodule showed malignant epithelial cells with features consistent with breast carcinoma in a background of benign thyroid epithelial cells and colloid. The case was signed out as metastatic breast carcinoma. Subsequent FNA and biopsy of her right shoulder lesion also revealed metastatic breast carcinoma with similar morphology to the material in the thyroid FNA.

Molecular profiling and classification of sporadic renal cell carcinoma by quantitative methylation analysis.

PURPOSE: Preoperative histologic classification of solid renal masses remains limited with current technology. We determine the utility of molecular profiling based on quantitative methylation analysis for characterization of sporadic renal cell carcinoma. EXPERIMENTAL DESIGN: Primary renal cell carcinomas representing three different histologic subtypes were obtained from a total of 38 patients who underwent radical nephrectomy for suspected malignant disease. Genomic DNA was isolated from tumors and was subjected to sodium bisulfite modification. The normalized index of methylation (NIM) for each sample was determined by quantitative real-time methylation-specific PCR at 17 different gene promoters. Hierarchical cluster analysis was performed by using an unsupervised neural network with binary tree topology. RESULTS: The majority of gene promoters that were analyzed in this study demonstrated very low levels of methylation (NIM <1.0). The RASSF1A gene promoter, however, was methylated in 30 of 38 (79%) cases. The frequency of RASSF1A methylation in papillary, clear-cell, and oncocytoma subtypes was 100, 90, and 25%, respectively. The highest levels of RASSF1A methylation were observed in the papillary (mean NIM = 78.9) and clear-cell (mean NIM = 13.4) subtypes. The vast majority of oncocytomas were completely unmethylated, and none demonstrated >1% methylation (mean NIM = 0.11). Hierarchical cluster analysis based on quantitative methylation levels resulted in stratification of sporadic renal cell carcinomas into their discrete histologic subtypes. CONCLUSIONS: Classification of sporadic renal cell carcinomas into histologic subtypes can be accomplished via multigene quantitative methylation profiling. Validation of this approach and selection of appropriate methylation markers may ultimately lead to use of this technology in the preoperative assessment of suspicious renal masses.

Nonrandom distribution of aberrant promoter methylation of cancer-related genes in sporadic breast tumors.

PURPOSE: In an effort to additionally determine the global patterns of CpG island hypermethylation in sporadic breast cancer, we searched for aberrant promoter methylation at 10 gene loci in 54 primary breast cancer and 10 breast benign lesions. EXPERIMENTAL DESIGN: Genomic DNA sodium bisulfate converted from benign and malignant tissues was used as template in methyl-specific PCR for BRCA1, p16, ESR1, GSTP1, TRbeta1, RARbeta2, HIC1, APC, CCND2, and CDH1 genes. RESULTS: The majority of the breast cancer (85%) showed aberrant methylation in at least 1 of the loci tested with half of them displaying 3 or more methylated genes. The highest frequency of aberrant promoter methylation was found for HIC1 (48%) followed by ESR1 (46%), and CDH1 (39%). Similar methylation frequencies were detected for breast benign lesions with the exception of the CDH1 gene (P = 0.02). The analysis of methylation distribution indicates a statistically significant association between methylation of the ESR1 promoter, and methylation at CDH1, TRbeta1, GSTP1, and CCND2 loci (P < 0.03). Methylated status of the BRCA1 promoter was inversely correlated with methylation at the RARbeta2 locus (P < 0.03). CONCLUSIONS: Our results suggest a nonrandom distribution for promoter hypermethylation in sporadic breast cancer, with tumor subsets characterized by aberrant methylation of specific cancer-related genes. These breast cancer subgroups may represent separate biological entities with potential differences in sensitivity to therapy, occurrence of metastasis, and overall prognosis.

Bipolar needle electrocautery for laparoscopic partial nephrectomy without renal vascular occlusion in a porcine model.

OBJECTIVES: To report a novel method of accomplishing laparoscopic lower pole partial nephrectomy in an acute porcine model using a bipolar needle electrode without the need for renal arterial occlusion. METHODS: Six animals (12 renal units) underwent laparoscopic polar nephrectomy using the bipolar needle electrode. After complete laparoscopic mobilization of the lower pole of the kidney, the bipolar needle electrode was repeatedly inserted full-thickness into the renal parenchyma and applied transversely, creating regional ischemia to the entire lower pole without renal vascular occlusion. The specimen was then amputated using laparoscopic scissors. RESULTS: For the 12 laparoscopic partial nephrectomies, the mean operative time was 39 +/- 30 minutes, and the mean blood loss was 90 +/- 112 mL. Of the 12 cases, 10 (83%) were performed successfully with the bipolar needle electrocautery as the only source of hemostasis and without the need for ancillary hemostatic measures. Two of the procedures (17%) required temporary arterial control for hemostasis. For the successful procedures, the mean operative time was 29 +/- 4 minutes, and the mean blood loss was 48 +/- 11 mL. Histologic analysis of the specimens demonstrated coagulative necrosis between 2 and 4 mm from the line of the surgical incision. CONCLUSIONS: Bipolar needle electrocautery is a promising device that can be used to facilitate laparoscopic partial nephrectomy with minimal blood loss and without the need for renal arterial occlusion and warm ischemia. Additional studies are required to optimize the delivery parameters of this device.

Molecular analysis of peritoneal fluid in ovarian cancer patients.

To determine whether genetic abnormalities present in primary ovarian tumors can be used to detect cancer cells in peritoneal fluid, we tested 14 ovarian cancers and 1 benign tumor of the ovary for loss of heterozygosity (LOH) at chromosomal arms 13q, 17p, 17q, and 22q and for mutations in the p53 and K-ras genes. In each case, matched primary tumor, normal tissue, and peritoneal fluid were analyzed. The highest frequency of LOH was found on chromosomal arm 17p (42%), followed by chromosomal arm 17q (36%), 22q (30%), and 13q (21%). Identical alterations were detected in matched peritoneal fluid (either peritoneal wash or ascitic fluid) in 3 of the 8 patients with LOH in the tumor (38%). Direct sequence analysis detected p53 mutations in 3 of the 14 malignant tumors (21%) and no (0) K-ras mutations. Identical mutations were detected in matched peritoneal fluid from all 3 patients with p53 mutations. All 8 of the 14 (57%) malignant tumors that showed at least one genetic abnormality were serous adenocarcinoma and identical alterations were detected in 5 of the 8 (62%) matched peritoneal fluid samples. Our findings indicate that molecular abnormalities can be detected in peritoneal fluid from patients with ovarian cancer and may be used to complement current conventional diagnostic procedures for detection of primary ovarian cancer.

Histologic evolution of high-intensity focused ultrasound in rabbit muscle.

RATIONALE AND OBJECTIVES: The purpose of this study was to examine the histologic evolution over time of rabbit skeletal muscle thermally ablated with high-intensity focused ultrasound. The objectives included determining the extent and focality of damage created by this noninvasive, transcutaneous ablative technology. METHODS: Transcutaneous, thermal ablation with an external focused ultrasound transducer was applied to the paraspinous muscles of 19 rabbits. At varying times, up to 100 days after therapy, single sonications were examined histologically. RESULTS: Initially, only subtle staining changes were identified within lesions. In the chronic phase (day 51-100), the muscle was replaced or infiltrated by variable amounts of scar and fat similar to degenerative muscle disorders. Histologic changes were limited to the tissue within the intensity focus of the transducer and were not seen in intervening tissues. DISCUSSION: The current study took a systematic approach to study the long term, in vivo histologic effects of single HIFU lesions in a nonregenerative tissue. This experience in muscle tissue will provide a basis for understanding ultrasound effects for clinical applications such as treatment of uterine fibroids, cardiac tissue, and sarcomas.

High-intensity focused ultrasound ablation of the epididymis in a canine model: a potential alternative to vasectomy.

BACKGROUND AND PURPOSE: High-intensity focused ultrasound (HIFU) is a noninvasive technology capable of inducing thermal coagulative necrosis of subsurface structures without injuring intervening tissues. We have previously reported on the feasibility of HIFU vasectomy in a canine model. In this study, we evaluated the feasibility of HIFU ablation of the canine epididymis as an alternative to vasectomy. The epididymis may be a better target than the vas deferens because it is larger, more easily positioned in the HIFU focal zone, and more susceptible to occlusive injury at lower energy levels, thus reducing collateral damage. MATERIALS AND METHODS: A hand-held HIFU clamp was used to grasp the epididymides of anesthetized dogs (eight surgically exposed and six grasped transcutaneously). An ultrasound transducer in the clamp focused energy on a 1 x 3 x 8-mm target zone centered between the clamp jaws. Ultrasonic energy (6-19 W) was delivered to this target zone for various times (16-150 seconds). The vas, epididymis, and testis were harvested for histologic examination 2 weeks after ablation. RESULTS: Seven of the eight epididymides ablated after surgical exposure demonstrated histologic findings associated with occlusion (fibrosis, hemorrhage, and proximal duct dilatation). Five of the six epididymides ablated transcutaneously also demonstrated histologic evidence of occlusion. Skin burns were evident overlying three epididymides, and one testicular injury was noted adjacent to an ablated region of the epididymis. CONCLUSION: High-intensity focused ultrasound ablation of the epididymis causes injury and histologic changes associated with epididymal occlusion. Further investigations are under way to optimize ablation parameters and to confirm azoospermia with ejaculate studies. Refinement of this technology may provide a rapid noninvasive alternative to conventional vasectomy.