Ischemic stroke alters immune cell niche and chemokine profile in mice independent of spontaneous bacterial infection.

INTRODUCTION: Stroke-associated pneumonia (SAP) is a major cause of mortality in patients who have suffered from severe ischemic stroke. Although multifactorial in nature, stroke-induced immunosuppression plays a key role in the development of SAP. Previous studies using a murine model of transient middle cerebral artery occlusion (tMCAO) have shown that focal ischemic stroke induction results in functional defects of lymphocytes in the spleen, thymus, and peripheral blood, leading to spontaneous bacterial infection in the lungs without inoculation. However, how ischemic stroke alters immune cell niche and the expression of cytokines and chemokines in the lungs has not been fully characterized. METHODS: Ischemic stroke was induced in mice by tMCAO. Immune cell profiles in the brain and the lungs at 24- and 72-hour time points were compared by flow cytometric analysis. Cytokine and chemokine expression in the lungs were determined by multiplex bead arrays. Tissue damage and bacterial burden in the lungs following tMCAO were evaluated. RESULTS: Ischemic stroke increases the percentage of alveolar macrophages, neutrophils, and CD11b+ dendritic cells, but reduces the percentage of CD4+ T cells, CD8+ T cells, B cells, natural killer cells, and eosinophils in the lungs. The alteration of immune cell niche in the lungs coincides with a significant reduction in the levels of multiple chemokines in the lungs, including CCL3, CCL4, CCL5, CCL17, CCL20, CCL22, CXCL5, CXCL9, and CXCL10. Spontaneous bacterial infection and tissue damage following tMCAO, however, were not observed. CONCLUSION: This is the first report to demonstrate a significant reduction of lymphocytes and multiple proinflammatory chemokines in the lungs following ischemic stroke in mice. These findings suggest that ischemic stroke directly impacts pulmonary immunity.

Trends in Bone Marrow Sampling and Core Biopsy Specimen Adequacy in the United States and Canada: A Multicenter Study.

OBJECTIVES: To assess bone marrow (BM) sampling in academic medical centers. METHODS: Data from 6,374 BM samples obtained in 32 centers in 2001 and 2011, including core length (CL), were analyzed. RESULTS: BM included a biopsy (BMB; 93%) specimen, aspirate (BMA; 92%) specimen, or both (83%). The median (SD) CL was 12 (8.5) mm, and evaluable marrow was 9 (7.6) mm. Tissue contraction due to processing was 15%. BMB specimens were longer in adults younger than 60 years, men, and bilateral, staging, and baseline samples. Only 4% of BMB and 2% of BMB/BMA samples were deemed inadequate for diagnosis. BM for plasma cell dyscrasias, nonphysician operators, and ancillary studies usage increased, while bilateral sampling decreased over the decade. BM-related quality assurance programs are infrequent. CONCLUSIONS: CL is shorter than recommended and varies with patient age and sex, clinical circumstances, and center experience. While pathologists render diagnoses on most cases irrespective of CL, BMB yield improvement is desirable.

Selective bladder denervation for overactive bladder (OAB) syndrome: From concept to healing outcomes using the ovine model.

AIMS: We evaluated a Selective Bladder Denervation (SBD) device, which uses radiofrequency ablation, for the treatment of overactive bladder syndrome in terms of its nerve denervation, ablation characteristics, and post-treatment healing. METHODS: Using the SBD device, eight fresh extirpated ovine bladder trigones were treated (90°C set point for 60 s) and nitroblue tetrazolium viability stained to characterize the ablation. In addition, 12 trigones were treated in vivo with three adjacent ablations and divided into survival cohorts: Day 7, Day 30, and Day 90 to assess the ablations and their associated healing. RESULTS: The ex vivo single trigone ablations had a 7.9 ± 0.9 mm width and 5.7 ± 1.0 mm thickness that involved the submucosa, detrusor muscle, adventitia, and vagina. Microscopic viability staining confirmed complete nerve necrosis within the targeted tissue. The in vivo Day 7 trigones supported the ex vivo ablation characteristics and showed up to minimal inflammation, granulation tissue, and collagen fibrosis. Day 30 trigones had essentially absent inflammation and granulation tissue with evolving collagen fibrosis at the ablation's periphery. Day 90 trigones had essentially absent acute inflammation, minimal chronic inflammation, essentially absent granulation tissue, and up to mild collagen fibrosis. No ureteral/urethral alterations, vesico-vaginal fistulas, or other complications were identified. CONCLUSIONS: The SBD device provided a targeted trigone ablation with resultant denervation. The tissue healing timeline followed that expected for a hyperthermic ablation and was characterized by a fibroproliferative healing response with limited inflammation and granulation tissue. The ablations did not impact the overlying bladder mucosal surface.

Metered Cryospray™: a novel uniform, controlled, and consistent in vivo application of liquid nitrogen cryogenic spray.

In this article, a novel cryotherapy approach using a uniform, controlled, and consistent in vivo application of liquid nitrogen (LN2) spray as a Metered Cryospray™ (MCS) process is described. Although MCS may be used for many potential clinical applications, this paper focuses on the development that led to the controlled and consistent delivery of radial LN2 cryogen spray in order to generate a uniform circumferential effect and how the amount of MCS can be adapted to specifically ablate targeted diseases within a patient's lumen such as an airway or esophagus.

Myeloid Sarcoma of the Bladder in the Setting of Refractory Anemia with Excess Blasts-2 (RAEB-2).

Myeloid sarcoma is an extramedullary tumor consisting of immature hematopoietic cells of granulocytic or monocytic differentiation. While rare, it can be seen in a variety of clinical settings and is most commonly associated with acute myeloid leukemia (AML), myelodysplastic syndromes (MDS) and myeloproliferative neoplasms (MPN). We present a rare case of myeloid sarcoma occurring in the bladder of a 56 year old male. Myeloid sarcoma may be difficult to recognize due to its rarity and clinical and morphologic similarity to many other conditions; however, swift diagnosis is necessary as it is considered equivalent to AML. Prognostic indicators for myeloid sarcoma have not been well established, but survival may be improved by undergoing chemotherapy designed to treat AML.

RepSox slows decay of CD34+ acute myeloid leukemia cells and decreases T cell immunoglobulin mucin-3 expression.

Despite initial response to therapy, most acute myeloid leukemia (AML) patients relapse. To eliminate relapse-causing leukemic stem/progenitor cells (LPCs), patient-specific immune therapies may be required. In vitro cellular engineering may require increasing the "stemness" or immunogenicity of tumor cells and activating or restoring cancer-impaired immune-effector and antigen-presenting cells. Leukapheresis samples provide the cells needed to engineer therapies: LPCs to be targeted, normal hematopoietic stem cells to be spared, and cancer-impaired immune cells to be repaired and activated. This study sought to advance development of LPC-targeted therapies by exploring nongenetic ways to slow the decay and to increase the immunogenicity of primary CD34(+) AML cells. CD34(+) AML cells generally displayed more colony-forming and aldehyde dehydrogenase activity than CD34(-) AML cells. Along with exposure to bone marrow stromal cells and low (1%-5%) oxygen, culture with RepSox (a reprogramming tool and inhibitor of transforming growth factor-ß receptor 1) consistently slowed decline of CD34(+) AML and myelodysplastic syndrome (MDS) cells. RepSox-treated AML cells displayed higher CD34, CXCL12, and MYC mRNA levels than dimethyl sulfoxide-treated controls. RepSox also accelerated loss of T cell immunoglobulin mucin-3 (Tim-3), an immune checkpoint receptor that impairs antitumor immunity, from the surface of AML and MDS cells. Our results suggest RepSox may reduce Tim-3 expression by inhibiting transforming growth factor-ß signaling and slow decay of CD34(+) AML cells by increasing CXCL12 and MYC, two factors that inhibit AML cell differentiation. By prolonging survival of CD34(+) AML cells and reducing Tim-3, RepSox may promote in vitro immune cell activation and advance development of LPC-targeted therapies.

Successful vapor-based endometrial ablation: in vivo peri-hysterectomy study.

STUDY OBJECTIVE: To evaluate the AEGEA vapor-based endometrial ablation system using an in vivo peri-hysterectomy model. DESIGN: Single-site feasibility study (Canadian Task Force classification II-2). SETTING: University medical center. PATIENTS: Nine women consented to undergo AEGEA endometrial ablation before previously scheduled abdominal hysterectomy to treat abnormal uterine bleeding. INTERVENTIONS: In vivo AEGEA endometrial ablation was performed using a 90-second vapor treatment cycle. After hysterectomy, the uteri were examined for the extent and location of endomyometrial ablation (macroscopic triphenyltetrazolium chloride staining) and fallopian tube injury (microscopic nitroblue tetrazolium staining). MEASUREMENTS AND MAIN RESULTS: The mean (SD) posttreatment measurements of the 9 uteri were as follows: weight, 143 (40) g; length, 10.3 (1.3 cm); thickness, 4.4 (0.6) cm; and width, 6.2 (0.7) cm. The endometrial thickness was 1.1 (0.7) mm. Three uteri had myomas that measured less than 2 cm; and 2 uteri demonstrated focal adenomyosis. No myometrial perforation or thermal serosal injury was identified. The median corpus, lower uterine cavity and bilateral cornua percentages of TTC-negative surface endometrial treatment were 100% (range: 100-100%), 100% (range: 80-100%), and 100% (range: 95-100%), respectively. The closest distance between the ablation and serosa was 11.5 (3.2) mm. No lower endocervical or exocervical thermal injury was identified. Minimal fallopian tube thermal injury was identified in 18% of interstitial segments evaluated, and measured 0.6 to 0.8 mm in maximal depth and extended to within 6.3 to 9.5 mm of the serosa. No thermal injury was identified in the extrauterine fallopian tube segments. CONCLUSION: The AEGEA vapor-based endometrial ablation system has the potential to provide excellent cavity coverage with full-thickness endometrial ablation. The study results further support an acceptable in vivo safety profile for future clinical efficacy trials.

AFAP1L1 is a novel adaptor protein of the AFAP family that interacts with cortactin and localizes to invadosomes.

The actin-filament associated protein (AFAP) family of adaptor proteins consists of three members: AFAP1, AFAP1L1, and AFAP1L2/XB130 with AFAP1 being the best described as a cSrc binding partner and actin cross-linking protein. A homology search of AFAP1 recently identified AFAP1L1 which has a similar sequence, domain structure and cellular localization; however, based upon sequence variations, AFAP1L1 is hypothesized to have unique functions that are distinct from AFAP1. While AFAP1 has the ability to bind to the SH3 domain of the nonreceptor tyrosine kinase cSrc via an N-terminal SH3 binding motif, it was unable to bind cortactin. However, the SH3 binding motif of AFAP1L1 was more efficient at interacting with the SH3 domain of cortactin and not cSrc. AFAP1L1 was shown by fluorescence microscopy to decorate actin filaments and move to punctate actin structures and colocalize with cortactin, consistent with localization to invadosomes. Upon overexpression in A7r5 cells, AFAP1L1 had the ability to induce podosome formation and move to podosomes without stimulation. Immunohistochemical analysis of AFAP1L1 in human tissues shows differential expression when contrasted with AFAP1 with localization of AFAP1L1 to unique sites in muscle and the dentate nucleus of the brain where AFAP1 was not detectable. We hypothesize AFAP1L1 may play a similar role to AFAP1 in affecting changes in actin filaments and bridging interactions with binding partners, but we hypothesize that AFAP1L1 may forge unique protein interactions in which AFAP1 is less efficient, and these interactions may allow AFAP1L1 to affect invadosome formation.

A Polymorphic Variant of AFAP-110 Enhances cSrc Activity.

Enhanced expression and activity of cSrc are associated with ovarian cancer progression. Generally, cSrc does not contain activating mutations; rather, its activity is increased in response to signals that affect a conformational change that releases its autoinhibition. In this report, we analyzed ovarian cancer tissues for the expression of a cSrc-activating protein, AFAP-110. AFAP-110 activates cSrc through a direct interaction that releases it from its autoinhibited conformation. Immunohistochemical analysis revealed a concomitant increase of AFAP-110 and cSrc in ovarian cancer tissues. An analysis of the AFAP-110 coding sequence revealed the presence of a nonsynonymous, single-nucleotide polymorphism that resulted in a change of Ser403 to Cys403. In cells that express enhanced levels of cSrc, AFAP-110(403C) directed the activation of cSrc and the formation of podosomes independently of input signals, in contrast to wild-type AFAP-110. We therefore propose that, under conditions of cSrc overexpression, the polymorphic variant of AFAP-110 promotes cSrc activation. Further, these data indicate amechanismby which an inherited genetic variation could influence ovarian cancer progression and could be used to predict the response to targeted therapy.

VE-cadherin and PECAM-1 enhance ALL migration across brain microvascular endothelial cell monolayers.

OBJECTIVE: Infiltration of the central nervous system (CNS) by leukemia is a problematic disease manifestation of acute lymphoblastic leukemia (ALL). The mechanisms by which leukocytes interact with human brain-derived microvasculature endothelial cells (HBMEC) and enter the CNS are largely derived from models of inflammation. However, our data indicate that ALL cells do not elicit an inflammatory phenotype by HBMEC. Our current investigation focuses on the contribution of the unique coexpression of vascular endothelial (VE)-cadherin and platelet endothelial cell adhesion molecule-1 (PECAM-1) by ALL in mediating leukemic cell interactions with HBMEC as an in vitro model of the blood-brain barrier. MATERIALS AND METHODS: Primary ALL and ALL cell lines were evaluated for VE-cadherin and PECAM-1 expression. Lentiviral-mediated transduction of VE-cadherin and PECAM-1 into REH cells and antibody neutralization of VE-cadherin and PECAM-1 in SUP-B15 cells was used to delineate the role of these two proteins in mediating ALL adhesion to, and migration through, HBMEC monolayers. RESULTS: Although cell line models indicate that VE-cadherin and PECAM-1 expression is found on the surface Philadelphia chromosome-positive ALL, evaluation of primary ALL demonstrates that VE-cadherin and PECAM-1 are expressed independent of Philadelphia status. Expression of VE-cadherin and PECAM-1 by ALL enhanced the adhesion of ALL to HBMEC, while expression of PECAM-1 enhanced ALL adhesion to, and migration through, HBMEC. CONCLUSIONS: Expression of VE-cadherin and PECAM-1 by ALL cells positions them to interact with HBMEC. By increasing our understanding of molecular mechanisms through which ALL cells gain entry into the CNS, new strategies may be designed to prevent leukemia cell entry into the CNS.

Greenlight photoselective 120-watt 532-nm lithium triborate laser vaporization prostatectomy in living canines.

PURPOSE: Efficiency, anatomic, and histopathologic outcomes of GreenLight HPS 120-W, 532-nm lithium triborate (LBO) laser photoselective vaporization of the prostate (PVP) in a survival model of living canines were studied and compared with the outcomes of the only benchmarked survival study of 60-W 532-nm potassium-titanyl-phosphate (KTP) laser PVP in living canines. MATERIALS AND METHODS: Twelve dogs underwent anterograde PVP with the 120-W LBO laser and, 4 each, were euthanized 3 hours (acute), 3 days (early), or 8 weeks (chronic) postoperatively. Laser energy and time were recorded. Prostates were sectioned, measured, and histologically analyzed after hematoxylin and eosin (H&E), triphenyltetrazolium chloride (TTC), or Gomori trichrome (GT) staining and compared with a normal control. RESULTS: LBO laser PVP at 120 W acutely created a 6.7 +/- 3.2 cm(3) cavity, hemostatically, and vaporized tissue 160% more efficiently (mean 1.3 cm(3)/min vs 0.5 cm(3)/min), 500% faster (mean 4.9 vs 29.1 min), and needed 121% less energy (mean 28.8 vs 63.6 kJ) than the 60-W KTP laser. Histologic staining with H&E and TTC demonstrated a coagulation zone of 1.5 +/- 0.3 mm for the 120-W LBO laser, comparable to the 1 to 2 mm for the 60-W KTP laser. H&E- and GT-stained, healed prostates at 8-weeks postoperatively showed reepithelialized cavities with minimal submucosal fibrosis compared with an identically stained normal and the benchmarked KTP laser PVP-treated prostates. CONCLUSION: Our in vivo canine survival study demonstrates GreenLight HPS 120-W, 532-nm LBO laser PVP has substantially more vaporization efficiency and speed, with equally favorable tissue interaction and healing vis-à-vis those benchmarked for the 532-nm wavelength by KTP laser PVP.

Comparison of four energy-based vascular sealing and cutting instruments: A porcine model.

AIM: To compare the safety and efficacy of four energy-based vascular sealing and cutting instruments. METHODS: Blood vessels of various types and diameters were harvested from four pigs using four instruments: Harmonic ACE (Ethicon Endo-Surgery, Cincinnati, OH), LigaSure V and LigaSure Atlas (Valleylab, Inc., Boulder, CO; a division of Tyco Healthcare), and EnSeal vessel fusion system (SurgRx, Inc. Redwood City, CA). The diameters of the vessels, speed and adequacy of the cutting and sealing process, and bursting pressures were compared. An additional set of specimens was sealed and left in situ for up to 4 h after which the vessels were harvested and histopathologically analyzed for the degree of thermal injury. RESULTS: The bursting pressures were significantly higher with EnSeal compared to all other instruments (p < 0.0001). The sealing process was significantly shorter with Harmonic ACE and significantly longer with LigaSure Atlas (p <0.0001). The mean seal width was larger with the LigaSure Atlas compared to the other instruments, and it was smaller with EnSeal and Harmonic ACE. Less radial adventitial collagen denaturation was present with EnSeal and LigaSure V than with the other two instruments; there were no significant differences in collagen denaturation although proximal thermal injury to the smooth muscle in the media of the vessel wall was less common with LigaSure Atlas than with the other instruments; however, the numbers were too small for statistical analysis. CONCLUSIONS: The bursting pressures with EnSeal were significantly higher than with all the other instruments. Harmonic ACE was the fastest sealing instrument and LigaSure Atlas was slowest. EnSeal created less radial thermal damage to the adventitial collagen of the vessels and LigaSure Atlas created less thermal damage to the media of the vessels. The clinical significance of these findings is unknown.

Biliary cystadenomas: hormone receptor expression and clinical management.

Biliary cystadenomas with mesenchymal stroma are neoplasms whose growth may be hormone sensitive. This study profiled the immunohistochemistry of these lesions to clarify the pathophysiology and define clinical management. Twelve patients with biliary cystadenomas were identified. Tissue was tested with a panel of probes including estrogen and progesterone receptors and compared to pancreatic and ovarian cystadenomas. Epithelial ER, PR, CD117, or SMA expression was negative in all three tumors. Epithelial CD10 expression was seen in 60% biliary, 75% pancreatic, and 0% ovarian tumors. Biliary cystadenoma stromal expression was ER+ (70%), PR+ (60%), CD10+ (40%), and c-kit+ (0%). Symptoms were seen in 92% patients. Percutaneous sclerotherapy and incomplete resection were associated with recurrence. Enucleation was the least morbid surgical technique. A role for hormonally mediated growth of biliary cystadenomas occurring through the stroma, rather than the epithelium, is suggested. Management remains complete surgical resection.

Molecular characterization of an 11q interstitial deletion in a patient with the clinical features of Jacobsen syndrome.

The 11q terminal deletion disorder or Jacobsen syndrome is a contiguous gene disorder. It is characterized by psychomotor retardation, cardiac defects, blood dyscrasias (Paris-Trousseau syndrome) and craniofacial anomalies. We report on a female patient with an approximately 10 Mb interstitial deletion with many of the features of Jacobsen syndrome: A congenital heart defect, dysmorphic features, developmental delay, and Paris-Trousseau syndrome. The karyotype of the patient is 46,XX,del(11)(q24.1q24.3). The interstitial deletion was confirmed using FISH probes for distal 11q, and the breakpoints were characterized by microarray analysis. This is the first molecularly characterized interstitial deletion in a patient with the clinical features of Jacobsen syndrome. The deletion includes FLI-1, but not JAM-3, which will help to determine the critical genes involved in this syndrome.

Posttransplant lymphoproliferative disorder in pancreas transplantation: a single-center experience.

BACKGROUND: Posttransplant lymphoproliferative disorder (PTLD) is a rare, serious complication of transplantation. The characteristics and associations of this disease in pancreas recipients have not been extensively studied. METHODS: From January 1988 through December 2002, 787 pancreas and 569 kidney-pancreas transplants were performed at our institution. Eighteen pancreas recipients developed polymorphic PTLD or malignant lymphoma. Data on clinical course, organ involvement, molecular characteristics, and association with immunosuppression and recent cytomegalovirus (CMV) infection were compiled from the institutional transplant database. Patient survival was compared to recipients of liver and kidney transplants at the same center by using Kaplan-Meier analysis. RESULTS: The 5-year cumulative incidence of PTLD in simultaneous pancreas-kidney, pancreas after kidney, and pancreas transplant alone recipients was 2.5%, 1.2%, and 1.0%, respectively (P = 0.23). A noticeably, but not significantly, higher cumulative incidence was seen in the more recent era since 1995 (2.1% vs. 0.9%, P = 0.15). PTLD in pancreas recipients carried a worse prognosis than in liver or kidney for recipients B-cell, early-onset, and Epstein Barr virus-positive lesions. PTLD was more aggressive in pancreas recipients, with a higher stage at presentation and a trend to more bone marrow involvement. There appeared to be a tendency toward association with recent CMV infection. Since 1995, PTLD recipients have had a lower exposure to antilymphocyte preparations (25 +/- 5 vs. 10 +/- 0.8)(P < 0.05). CONCLUSIONS: PTLD in pancreas recipients remains a rare but aggressive disease, and carries a worse prognosis in comparison to other transplant recipients. These heavily immunosuppressed patients, who often face multiple transplants, may be at greater risk; CMV infection may play an antecedent role.

A cryoinjury model using engineered tissue equivalents for cryosurgical applications.

Cryosurgery is emerging as a promising treatment modality for various cancers, but there are still challenges to be addressed to improve its efficacy. Two primary challenges are determining thermal injury thresholds for various types of cell/tissue, and understanding of the mechanisms of freezing induced cell/tissue injury within a cryolesion. To address these challenges, various model systems ranging from cell suspensions to three-dimensional in vivo tissues have been developed and used. However, these models are either oversimplifications of in vivo tissues or difficult to control and extract precise experimental conditions from. Therefore, a more readily controllable model system with tissue-like characteristics is needed. In this study, a cryoinjury model was developed using tissue engineering technology, and the capabilities of the model were demonstrated. Engineered tissue equivalents (TEs) were constructed by seeding and culturing cells in a type I collagen matrix. Two different cell lines were used in this study, AT-1 rat prostate tumor cells and LNCaP human prostate cancer cells. The constructed TEs underwent a freeze/thaw cycle imitating in vivo cryosurgery. Thermal conditions within TEs during freeze/thaw cycles were characterized, and the responses of TEs to these thermal conditions including freezing induced cellular injury and extracellular matrix damage were investigated at three different time points. The results illustrate the feasibility to establish thermal thresholds of cryoinjury for different cell/tissue types using the presently developed model, and its potential capabilities to study cell death mechanisms, cell proliferation or migration, and extracellular matrix structural damage after a freeze/thaw cycle.

Low blast count myeloid disorders with Auer rods: a clinicopathologic analysis of 9 cases.

Auer rods are a hallmark of acute myeloid leukemia but occasionally are seen in myelodysplastic syndromes (MDSs) or chronic myelomonocytic leukemia, rarely in cases with fewer than 5% blasts. The significance of this finding is unclear. We report 9 cases of this unusual phenomenon. All patients had cytopenias, isolated to a single lineage in 4. Circulating blasts were present in 8 cases (rare to 2.5%). Bone marrow blasts ranged from 0.4 to 4.9%; 1% to 32% of blasts contained Auer rods. There were variable degrees of dysplasia; 1 case closely mimicked refractory anemia with ringed sideroblasts. Cytogenetic studies in 8 cases showed clonal changes in 4. In 5 patients, acute myelogenous leukemia (AML) developed 6, 6, 5, 13, and 24 months after diagnosis; the patients subsequently died. Three patients died at 1, 1, and 8 months without progression to AML, and only 1 was alive at 10 months. MDSs with fewer than 5% blasts and Auer rods seem to be a heterogeneous group, but rapid progression to death or AML in most cases suggests that Auer rods signify an aggressive biology in MDSs with a low blast count.

Fatal hemorrhage from androgen-related hepatic adenoma after hematopoietic cell transplantation.

Fanconi anemia is a rare genetic disorder that leads to bone marrow failure. Hematopoietic cell transplantation (HCT) is currently the only treatment option with curative potential. When a suitable HLA-matched sibling donor is not available, patients are often treated with androgenic steroids before considering HCT. Such androgen treatments can lead to the development of hepatic adenomas, which usually regress upon stopping androgen therapy. A patient with Fanconi anemia is described who underwent an unrelated umbilical cord blood transplant with a history of a hepatic adenoma related to androgen therapy. No adenomas were detected on an ultrasound examination prior to HCT. Soon after HCT, he died due to sudden rupture and hemorrhage of a hepatic adenoma. This case illustrates the need for extra vigilance in the detection and management of hepatic adenomas in patients treated with androgens, especially prior to HCT.

Radiofrequency ablation causes 'thermal fixation' of hepatocellular carcinoma: a post-liver transplant histopathologic study.

Radiofrequency ablation (RFA) is increasingly used to treat hepatocellular carcinoma (HCC) in patients awaiting a liver transplant. Despite its increasing use, detailed histologic information is scarce regarding the nature of RFA-treated lesions. We identified four chronic hepatitis C patients who had RFA of their HCC before their liver transplant. For these four patients, we conducted a detailed histopathologic analysis of the treated lesions in their explanted livers. The five lesions included immediate (4 d) and long-term (14 months) post-RFA specimens. Of the five lesions, four were completely ablated. The one incompletely ablated lesion was also treated with chemoembolization. In the acute post-RFA period, a zone of interstitial hemorrhage occurred at the outer boundary of the lesion. Differing from classic tissue necrosis, the treated lesions all showed 'thermal fixation', with preserved tissue architecture and microscopic cellular detail. The cellular staining characteristics faded with time, but the treated tissue became brittle, resisted tissue breakdown, and generated a minimal wound healing response. At the periphery of the lesion, the fibrous septae of the cirrhotic liver and vascular structures appeared to demarcate or limit progression of the ablation front. A narrow hypocellular fibrous boundary with a focal 'foreign body' giant cell-type reaction developed around the edge of the ablation zone. Thus, RFA can produce immediate and complete thermal fixation of select lesions with an appropriate liver margin and can provide a satisfactory treatment option for select HCC patients before a liver transplant.

Cryothermic and hyperthermic treatments of human leiomyomata and adjacent myometrium and their implications for laparoscopic surgery.

STUDY OBJECTIVE: To evaluate the effects and feasibility of direct cryothermic and hyperthermic therapy on leiomyomata and adjacent myometrium, and to contribute to evidence-based treatment thresholds based on measurements of direct cell injury. DESIGN: Experimental study (Canadian Task Force classification II-2). SETTING: University hospital. SUBJECTS: Leiomyoma and myometrium tissue from 10 women undergoing total abdominal hysterectomy with or without bilateral salpingo-oophorectomy. INTERVENTION: In vitro cryothermic or hyperthermic therapy was performed with representative leiomyoma and myometrium tissue samples. Using a directional solidification stage to simulate cryothermic therapy, 10 leiomyoma and 6 myometrium specimens were cooled in vitro at a rate of -5 degrees C/minute to end temperatures of -20 degrees, -40 degrees, -60 degrees, and -80 degrees C with a 15-minute hold period and then rapidly thawed to 21 degrees C. Hyperthermic therapy was simulated using a preheated 45 degrees, 55 degrees, 60 degrees, 65 degrees, 70 degrees, 75 degrees, and 80 degrees C constant temperature copper heating block with a 10-minute treatment period. In conjunction with tissue culturing and control tissues, cell death was assessed with routine histology and viability dyes (ethidium homodimer/Hoechst). MEASUREMENTS AND MAIN RESULTS: In cryothermic results, leiomyomata cell death (LCD) increased from 12% to 27% by histology and 26% to 38% by viability dye assay over the thermal range from -20 degrees to -80 degrees C, respectively. Myometrial cell death (MCD) increased from 10% to 12% and 4% to 20% for the same measurements, respectively. Whereas MCD appeared relatively stable from -40 degrees to -80 degrees C, it was significantly less than LCD over this range (p <0.05). For hyperthermic results, LCD increased from 17% to 88% by histology with progressive temperature increase from 45 degrees to 80 degrees C, respectively. The MCD showed a similar increase from 16% to 91% by histology over this temperature range. Hyperthermic histology and dye assay results were similar for LCD and MCD. CONCLUSIONS: In comparison with myometrium, leiomyomata showed greater direct cryothermic and equal hyperthermic cell injury. Whereas cell death increased up to 70 degrees C and down to -80 degrees C, the interval increases in cell injury diminished with more extreme temperatures. In vivo studies of combined direct and ischemic vascular injury thresholds have yet to be performed, but direct LCD matrixes determined in this study will help provide guidelines for minimally invasive surgical techniques for the treatment of leiomyomata.