Histological Necrosis Research Articles

La phonomyographie comme méthode non invasive de détection et de suivi du syndrome de loge – évaluation sur un modèle de syndrome de loge chez le rat

Introduction Phonomyography (PMG) is a noninvasive acoustic technique by which microphones placed on the skin surface detect the low-frequency sounds emitted by muscle upon contraction. Presently, there is no reliable noninvasive continuous monitoring technique for the detection of acute compartment syndrome (ACS), a traumatic condition seen in military and civilian trauma victims. The phase 1 of this study (ischemia induction of 15 adult rat legs through direct iliac vessel clamping) has already shown promising results with decrease in the signal amplitude of PMG proportional to ischemia duration. The goal of the phase 2 is to confirm the ability of PMG to detect early changes in muscle contraction during ACS in a rat model. Material Adult Sprague rats were used for the phase 2 of the experiment. Methods In 14 adult Sprague rats, after general anesthesia induction, an acute compartment syndrome was induced by infusion of a physiologic saline solution into the left posterior calf compartment (direct ACS induction model), while the right posterior calf compartment was used as a control. At intervals of 10 minutes, for a maximum of 6 hours, the sciatic nerve of each leg was stimulated with a nerve stimulator, and the PMG acoustic signal recorded. ACS duration was different for each group of 3 rats (30 min, 1 h, 2 h, 4 h and 6 h). At the end of the procedure, compartment was released with fasciotomy and rats awakened after skin closure. At day 4, calf muscle biopsies were harvested and sent to pathology. Results We were able to get data for 14 rats (one rat in the 6 h group died at the end of the procedure). The PMG showed a decrease in the amplitude of the low-frequency signal emitted from injured muscle of the ACS limb, which correlated with the duration of muscle and nerve injury ischemia and histologic necrosis. At time t = 4 h, there was a statistically significant ( P n = 5 + P = 0.043). Discussion These findings further confirm the results obtained during the phase 1 (ischemia induction through iliac vessel clamping) and the potential of PMG as a continuous noninvasive detection method for ACS, by showing an alteration in the acoustic signal emitted by the muscles of an injured ischemic limb. Conclusion PMG is a promising device (patented) for continuous noninvasive monitoring of suspected ACS. The next step is to evaluate its accuracy with patient operated on for leg fracture, compared to direct pressure measurement.

Prognostic Value of 18-Fluorodeoxyglucose-Positron Emission Tomography After Transarterial Chemoembolization in Patients With Hepatocellular Carcinoma Undergoing Orthotopic Liver Transplantation

IntroductionThe aim of this study was to analyze the correlation between 18-FDG positron emission tomography (PET)/computed tomography (CT), histological necrosis, and prognosis after transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) patients undergoing orthotopic liver transplantation (OLT). PatientsFrom January 2007 through December 2013, 81 patients with HCC and liver cirrhosis were included in our liver transplantation program. For this study we selected patients who underwent 18-FDG PET/CT pre-TACE and post-TACE. All patients underwent liver transplantation within 8 weeks after PET/CT. ResultsTwenty patients with a median age of 58 years (range, 46–69 years) underwent an 18-FDG PET/CT before and after TACE. The median Standardized Uptake Value (SUV) before TACE was 3.8 (range, 2.6–8.7), with a median post-TACE SUV of 0% (range, 0–4). Among patients whose post-TACE SUV decreased to <3, >70% necrosis was observed upon study of a hepatectomy sample, with a survival rate of 100% and 80% at 1 and 3 years, respectively. ConclusionIn conclusion, performance of an 18-FDG PET/CT before and after TACE with comparison of SUV values among patients with HCC awaiting OLT provided valuable information regarding the effectiveness of TACE.

Anti-inflammatory, Antithrombotic and Cardiac Remodeling Preventive Effects of Eugenol in Isoproterenol-Induced Myocardial Infarction in Wistar Rat.

This study aimed to evaluate the antithrombotic, anti-inflammatory and anti-cardiac remodeling properties of eugenol in isoproterenol-induced myocardial infarction in rats. Male Wistar rats were randomly divided into four groups, control, iso [100mg/kg body weight was injected subcutaneously into rats at an interval of 24h for 2days (6th and 7th day) to induce MI] and pretreated animals with clopidogrel (0.2mg/kg) and eugenol (50mg/kg) orally for 7days and intoxicated with isoproterenol (Iso+Clop) and (Iso+EG) groups. Isoproterenol-induced myocardial infarcted rats showed notable changes in the ECG pattern, increase in heart weight index, deterioration in the hemodynamic function and rise in plasma level of troponin-T, CK-MB and LDH and ALT by 316, 74, 172 and 45%, respectively, with histological myocardium necrosis and cells inflammatory infiltration. In addition, significant increases in plasma levels of inflammatory biomarkers such as fibrinogen, α1, α2, β1, β2 and γ globulins with decrease level of albumin were observed in infarcted rats as compared to normal ones. Else, the angiotensin-converting enzyme (ACE) activity in plasma, kidney and heart of the isoproterenol-induced rats was significantly increased by 34, 47 and 93%, respectively, as compared to normal group. However, the administration of eugenol induced a clear improvement in cardiac biomarkers injury, reduced inflammatory mediators proteins, increased heart activities of superoxide dismutase and glutathione peroxidase with reduce in thiobarbituric acid-reactive substances content and inhibition of ventricular remodeling process through inhibition of ACE activity. Overall, eugenol evidences high preventive effects from cardiac remodeling process.

Imaging the Met Receptor Tyrosine Kinase (Met) and Assessing Tumor Responses to a Met Tyrosine Kinase Inhibitor in Human Xenograft Mouse Models with a [ 99m Tc] (AH-113018) or CY 5** (AH-112543) Labeled Peptide

Developing an imaging agent targeting the hepatocyte growth factor receptor protein (Met) status of cancerous lesions would aid in the diagnosis and monitoring of Met-targeted tyrosine kinase inhibitors (TKIs). A peptide targeting Met labeled with [(99m)Tc] had high affinity in vitro (Kd = 3.3 nM) and detected relative changes in Met in human cancer cell lines. In vivo [(99m)Tc]-Met peptide (AH-113018) was retained in Met-expressing tumors, and high-expressing Met tumors (MKN-45) were easily visualized and quantitated using single-photon emission computed tomography or optical imaging. In further studies, MKN-45 mouse xenografts treated with PHA 665752 (Met TKI) or vehicle were monitored weekly for tumor responses by [(99m)Tc]-Met peptide imaging and measurement of tumor volumes. Tumor uptake of [(99m)Tc]-Met peptide was significantly decreased as early as 1 week after PHA 665752 treatment, corresponding to decreases in tumor volumes. These results were comparable to Cy5**-Met peptide (AH-112543) fluorescence imaging using the same treatment model. [(99m)Tc] or Cy5**-Met peptide tumor uptake was further validated by histologic (necrosis, apoptosis) and immunoassay (total Met, p Met, and plasma shed Met) assessments in imaged and nonimaged cohorts. These data suggest that [(99m)Tc] or Cy5**-Met peptide imaging may have clinical diagnostic, prognostic, and therapeutic monitoring applications.

AB090. Establishment of prediction model of progression after radical nephrectomy or partial nephrectomy for Chinese patients with clear cell renal cell carcinoma

ObjectiveTo develop an algorithm to predict progression to relapse or metastases after radical or partial nephrectomy for Chinese patients with localized clear cell RCC (ccRCC), so as to guide the postoperative treatment.MethodsThe clinical and pathological features and prognosis of 1,034 localized ccRCC patients between January 2006 and December 2013 in the second hospital of Tianjin Medical University was analyzed retrospectively. Univariate comparisons of survival analysis used the Kaplan-Meier method. COX regression model method was used in the multivariate analysis. Harrell’s concordance index was used to assess the prognostic accuracy of the new model.ResultsThe median follow-up was 39 months (range 4-109 months). Relapse or metastases occurred in 129 patients. The relapse-free survival (RFS) rates after 1 year, 3 years, 5 years, 7 years were 94.8%, 88.6%, 83.4% and 80.4% respectively. In these 1,034 ccRCC patients, 130 patients underwent partial nephrectomy and 904 patients underwent radical nephrectomy. There was no difference in RFS rates between group of partial nephrectomy and group of radical nephrectomy (P=0.061). Multivariate analysis showed that the features of age, tumor size, symptoms at presentation, preoperative platelet count, tumor stage [2010], Fuhrman grade, histologic tumor necrosis were independent predictors associated with RFS for ccRCC patients. A scoring algorithm to predict progression to relapse or metastases after patients underwent partial or radical nephrectomy for ccRCC was developed using the regression coefficients from the multivariate analysis. In this model, the score was calculated as 2 (for pT1b), 3 (for pT2), 4 (for pT3 and pT4), 1 (for age ≥58), 1 (for tumor size ≥5.5 cm), 1 (for symptoms at presentation), 2 (for platelet count ≥400×109/L), 1 (for grade 3), 3 (for grade 4), 1 (for histologic tumor necrosis) and 0 otherwise. The risk of progression to relapse or metastases was divided into three groups: low-risk group (score 0-3), intermediate-risk group (score 4-7) and high-risk group (score ≥8). The estimated 5-year RFS rates were 96.9%, 72.7% and 13.1% respectively in these three groups (P<0.001). Using this sample, the C-index of Leibovich model was 0.784. The C-index of the new model validation was 0.802.ConclusionsN patients with ccRCC, age, tumor size, presenting symptoms, preoperative platelet count, tumor stage [2010], Fuhrman grade and histologic tumor necrosis are significant independent predictors of RFS after surgery. Based on these indicators, we established a scoring algorithm that can be used to predict disease progression after surgery for Chinese patients with ccRCC.

Serum Adiponectin Predicts Cancer-specific Survival of Patients with Renal Cell Carcinoma

BackgroundPrediction of outcomes in patients with renal cell carcinoma (RCC) is crucial for clinical decision-making. The limited accuracy of conventional prognostic factors such as stage and grade may be increased by the use of biomarkers. ObjectiveTo evaluate the association of serum adiponectin and leptin and polymorphisms in the leptin and leptin receptor genes with RCC histopathology and prognosis. Design, setting, and participantsAdiponectin and leptin levels were measured in preoperative serum samples from 131 consecutive patients with sporadic unilateral RCC. The polymorphisms G–2548A (rs7799039) in the leptin gene (LEP) and Gln223Arg (Q223R, A668G, rs1137101) in the leptin receptor gene (LEPR) were genotyped in 233 patients. Outcome measurements and statistical analysisMultivariable associations with RCC-specific survival were analyzed using Cox models. Results and limitationsMedian preoperative serum adiponectin was 15.8μg/ml (interquartile range 10.0–23.1). Adiponectin was lower in patients with distant metastases (p=0.017) or histologic tumor necrosis (p=0.015). On multivariable analysis adjusted for the effects of variables in the Karakiewicz nomogram, each 1-μg/ml increase in adiponectin was associated with a 8% decrease in the hazard of death from RCC (hazard ratio 0.92, 95% confidence interval 0.86–0.98; p=0.007). The discrimination of the Karakiewicz nomogram increased by 0.6% on inclusion of adiponectin. Leptin levels, LEP G–2548A and LEPR Q223R were not associated with either RCC pathology or outcomes. Limitations include the retrospective study design, the low numbers of patients, and a lack of standardized follow-up. ConclusionsThis study suggests that lower preoperative serum adiponectin is associated with features of biologically aggressive RCC, metastasis, and survival. Patient summaryWe assessed the relationship between outcomes and blood levels of adiponectin and leptin and genetic changes in leptin and leptin receptor genes. We found that patients with lower adiponectin levels have more aggressive tumors and poorer survival.

Evaluation of static and dynamic MRI for assessing response of bone sarcomas to preoperative chemotherapy: Correlation with histological necrosis.

Objectives:Preoperative chemotherapy plays a key role in management of bone sarcomas. Postoperative evaluation of histological necrosis has been the gold standard method of assessing response to preoperative chemotherapy. This study was done to evaluate the efficacy of static and dynamic magnetic resonance imaging (MRI) for assessing response preoperatively.Materials and Methods:Our study included 14 patients (12 osteosarcomas and 2 malignant fibrous histiocytomas) with mean age of 21.8 years, treated with preoperative chemotherapy followed by surgery. They were evaluated with static and dynamic MRI twice, before starting chemotherapy and again prior to surgery. Change in tumor volume and slope of signal intensity - time curve were calculated and correlated with percentage of histological necrosis using Pearson correlation test.Results:The change in dynamic MRI slope was significant (P = 0.001). Also, ≥60% reduction in slope of the curve proved to be an indicator of good histological response [positive predictive value (PPV) =80%]. Change in tumor volume failed to show significant correlation (P = 0.071). Although it showed high negative predictive value (NPV = 85.7%), PPV was too low (PPV = 57.14%).Conclusions:Dynamic MRI correctly predicts histological necrosis after administration of preoperative chemotherapy to bone sarcomas. Hence, it can be used as a preoperative indicator of response to neoadjuvant chemotherapy. On the other hand, volumetric assessment by static MRI is not an effective predictor of histological necrosis. This study proves the superiority of dynamic contrast-enhanced study over volumetric study by MRI.

Intratumor cholesteryl ester accumulation is associated with human breast cancer proliferation and aggressive potential: a molecular and clinicopathological study.

BackgroundThe metabolic effect of intratumor cholesteryl ester (CE) in breast cancer remains poorly understood. The objective was to analyze the relationship between intratumor CE content and clinicopathological variables in human breast carcinomas.MethodsWe classified 30 breast carcinoma samples into three subgroups: 10 luminal-A tumors (ER+/PR+/Her2-), 10 Her-2 tumors (ER-/PR-/Her2+), and 10 triple negative (TN) tumors (ER-/PR-/Her2-). We analyzed intratumor neutral CE, free cholesterol (FC) and triglyceride (TG) content by thin layer chromatography after lipid extraction. RNA and protein levels of lipid metabolism and invasion mediators were analyzed by real time PCR and Western blot analysis.ResultsGroup-wise comparisons, linear regression and logistic regression models showed a close association between CE-rich tumors and higher histologic grade, Ki-67 and tumor necrosis. CE-rich tumors displayed higher mRNA and protein levels of low-density lipoprotein receptor (LDLR) and scavenger receptor class B member 1 (SCARB1). An increased expression of acetyl-Coenzyme A acetyltransferase 1 (ACAT1) in CE-rich tumors was also reported.ConclusionsIntratumor CE accumulation is intimately linked to proliferation and aggressive potential of breast cancer tumors. Our data support the link between intratumor CE content and poor clinical outcome and open the door to new antitumor interventions.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-015-1469-5) contains supplementary material, which is available to authorized users.

Prognostic factors and survival rate of osteosarcoma: A single-institution study.

Osteosarcoma is a highly malignant primary bone tumor. The study aim to evaluate the prognostic factors influencing the survival rate in our center. This was a retrospective cohort study of all patients treated between January 2005 and December 2010. We included 163 patients with an age range of 6-59 years (median = 19). The median follow-up was 47 months (range 36-84). The overall survival in patients who completed chemotherapy and surgery (n = 117) was 72% at 2 years and 44% at 5 years. Histologically, 99 (85%) had osteoblastic, 6 (5%) had chondroblastic and 3 (2.5%) had telangiectatic osteosarcoma. Limb salvage surgery was performed in 80 (49%) and 41 (25%) underwent amputation. However, 46 patients (28%) underwent no surgical intervention and incomplete chemotherapy. In total, 38/79 patients had a good chemotherapy response. There was a significantly better survival rate for limb salvage versus amputation. Independent prognostic factors for survival are compliance to treatment and presence of lung metastasis. The overall survival of osteosarcoma patients was influenced by the presence of pulmonary metastases and compliance to treatment. Histological subtype, different chemotherapy regimens and histological necrosis after chemotherapy did not significantly influence survival. The patients who did not complete treatment had significantly poorer survival.

Preoperative serum-gamma-glutamyltransferase (GGT) does not represent an independent prognostic factor in a European cohort of patients with non-metastatic renal cell carcinoma

AimsIncreasing evidence suggests that the serum-gamma-glutamyltransferase (GGT) might correlate with tumour development and growth rates in various human cancer types. Thus, we decided to investigate the potential prognostic impact of...

Application of IL-36 receptor antagonist weakens CCL20 expression and impairs recovery in the late phase of murine acetaminophen-induced liver injury.

Overdosing of the analgesic acetaminophen (APAP, paracetamol) is a major cause of acute liver injury. Whereas toxicity is initiated by hepatocyte necrosis, course of disease is regulated by mechanisms of innate immunity having the potential to serve in complex manner pathogenic or pro-regenerative functions. Interleukin (IL)-36γ has been identified as novel IL-1-like cytokine produced by and targeting epithelial (-like) tissues. Herein, we investigated IL-36γ in acute liver disease focusing on murine APAP-induced hepatotoxicity. Enhanced expression of hepatic IL-36γ and its prime downstream chemokine target CCL20 was detected upon liver injury. CCL20 expression coincided with the later regeneration phase of intoxication. Primary murine hepatocytes and human Huh7 hepatocellular carcinoma cells indeed displayed enhanced IL-36γ expression when exposed to inflammatory cytokines. Administration of IL-36 receptor antagonist (IL-36Ra) decreased hepatic CCL20 in APAP-treated mice. Unexpectedly, IL-36Ra likewise increased late phase hepatic injury as detected by augmented serum alanine aminotransferase activity and histological necrosis which suggests disturbed tissue recovery upon IL-36 blockage. Finally, we demonstrate induction of IL-36γ in inflamed livers of endotoxemic mice. Observations presented introduce IL-36γ as novel parameter in acute liver injury which may contribute to the decision between unleashed tissue damage and initiation of liver regeneration during late APAP toxicity.

Outcome prediction for patients with renal cell carcinoma.

Outcome assessment for renal cell carcinoma is somewhat controversial. Despite numerous studies, a very limited variety of features have been recognized as having prognostic significance in clinical practice. In this review, tumor features considered to be of importance in outcome prediction for surgically treated patients with the 3 most commonly encountered morphotypes of renal cell carcinoma (clear cell, papillary, and chromophobe renal cell carcinoma) are evaluated. In particular, we have focused upon histologic subtype, sarcomatoid and rhabdoid differentiation, TNM staging, primary tumor size, tumor grade, and the presence of histologic coagulative tumor necrosis. We have also examined the importance of these prognostic features in a variety of postoperative or outcome prediction models developed by several institutions.

The Preoperative AST/ALT (De Ritis) Ratio Represents a Poor Prognostic Factor in a Cohort of Patients with Nonmetastatic Renal Cell Carcinoma

Aminotransaminases, which are strongly involved in cellular metabolism and cancer cell turnover, represent easily measureable, potential blood based biomarkers. We evaluated the prognostic value of the preoperatively assessed AST/ALT (De Ritis) ratio on clinically meaningful end points in a large European cohort of patients with nonmetastatic renal cell carcinoma. We retrospectively evaluated clinicopathological data on 698 patients with nonmetastatic renal cell carcinoma operated on between 2005 and 2013 at a single tertiary academic center. The potential prognostic value of the AST/ALT ratio was analyzed using the Kaplan-Meier method, and univariate and multivariate Cox proportional regression models. The impact of the ratio on the predictive accuracy of the Leibovich prognosis score was determined by the Harrell c-index. An increased (1.26 or greater) preoperative AST/ALT ratio was statistically significantly associated with several well established prognostic factors, including pathological T stage, as well as with histological tumor necrosis (p <0.05). On multivariate analysis an increased preoperative AST/ALT ratio was an independent prognostic factor for metastasis-free survival (HR 1.61, 95% CI 1.25-2.07, p <0.001) and overall survival (HR 1.76, 95% CI 1.34-2.32, p <0.001). The Harrell c-index was 0.77 using the Leibovich prognosis score and 0.81 when AST/ALT was added. In our study cohort with nonmetastatic renal cell carcinoma the preoperatively assessed AST/ALT ratio represented an independent prognostic factor. This ratio might further improve the predictive accuracy of well established prognosis scores.

Radionécrose cérébrale après irradiation en conditions stéréotaxiques des berges opératoires : analyse de la littérature à partir de quatre cas

Stereotactic hypofractionated radiotherapy after resection of brain metastasis is an alternative to whole brain radiotherapy. A high dose per fraction is associated with a risk of radiation necrosis. We present four cases of confirmed histological radiation necrosis. Differentiating recurrent tumour from radiation necrosis in this scenario is challenging. An enhancing area in magnetic resonance imaging (MRI) with a “cut bell pepper” appearance may suggest radiation necrosis. Advanced imaging modalities such as perfusion MR imaging and positron emission tomography can be useful. Dosimetric predictors of the occurrence of radiation necrosis after stereotactic hypofractionated radiotherapy are poorly understood and require prospective studies on larger cohorts.

Local administration of zoledronic acid for giant cell tumor of bone.

Giant cell tumor of bone is a locally aggressive tumor with a high local recurrence rate. Several adjuvant therapies have been employed to reduce the recurrence rate, but their effectiveness remains controversial. The authors attempted local administration of zoledronic acid, a nitrogen-containing bisphosphonate that strongly inhibits bone resorption, as an adjuvant treatment for histologically proven giant cell tumor of bone in 5 patients at their institution. After biopsy, 4 patients were treated with local administration of zoledronic acid with artificial bone and 1 was treated with zoledronic acid without artificial bone. Histologic response to the treatment was evaluated with surgically resected specimens. The 4 patients treated with artificial bone showed local control, with histologic tumor necrosis rates of 90%, 90%, 50%, and 10%. Magnetic resonance imaging showed poor gadolinium enhancement, and histologic examination after local zoledronic acid treatment showed tumor necrosis. One patient without artificial bone showed no histologic tumor necrosis and had local recurrence in soft tissue 18 months after tumor resection. A 3-week waiting period between biopsy and zoledronic acid treatment appears reasonable from the histological study. Complication of this therapy was delayed wound healing and it occurred in 2 cases. Taken together, this case series suggests that local administration of zoledronic acid with artificial bone is a potential adjuvant therapy for giant cell tumor of bone. On the other hand, effective local administration of zoledronic acid requires some bone matrix, including artificial bone. Campanacci's grading is important for predicting the effect of local administration of zoledronic acid.

Evaluation of anti-tumor activity of Momordica cymbalaria fenzl

Background: The aim of the current study is to evaluate the anti-tumor activity of saponins isolated from the roots of Momordica cymbalaria (MC) against dimethylbenz[a]anthracene (DMBA) induced rats. Methods: A steroidal saponin MC (SMC) was isolated from MC fenzl and purified by preparative high-performance liquid chromatography. Breast cancer was induced in 50-day-old female Sprague-Dawley rats by injecting DMBA (6 mg/kg intravenous) in three doses on day 50, 54, and 57. The rats were randomized into four groups; control, DMBA, SMC (100 mg/kg), and tamoxifen (6.6 mg/kg) to DMBA breast cancer rats. The tumor size, volume, hormonal, antioxidant, and whole mount parameters were estimated. Results: Mean tumor size and volume, luteinizing hormone, and progesterone with superoxide dismutases, catalase, and glutathione levels increased significantly (p<0.001); serum estradiol, follicle stimulating hormone with lipid peroxidation decreased significantly (p<0.001) in DMBA-induced breast cancer and vice versa in SMC and tamoxifen. Terminal end buds, terminal ducts, alveolar buds, and lobules decreased significantly (p<0.001) in DMBA-induced breast cancer whereas increased significantly in SMC and tamoxifen. Histological necrosis and hemorrhage along with focal desmoplastic reaction in DMBA-induced breast cancer; ductile elongation and hyperplasia of both ducts and alveoli were prominent, with increased secretory activity in SMC group. The results confirmed the chemopreventive effect of SMC and tamoxifen in DMBA-induced breast cancer. Conclusions: The SMC exhibited anti-tumor activity against mammary cancer, which may be due to its anti-estrogenic, antioxidant activity.

Tumor necrosis is a strong predictor for recurrence in patients with pathological T1a renal cell carcinoma.

Patients with pT1aN0M0 renal cell carcinoma (RCC) generally have good prognosis, and recurrence is rare. However, metastasis develops postoperatively in a small number of patients with pT1aN0M0 RCC. The present study was undertaken to identify predictors for recurrence in patients with pT1aN0M0 RCC. We reviewed the clinicopathological factors of 133 patients with pT1aN0M0 RCC who underwent radical or partial nephrectomy at the Department of Urology, National Defense Medical College (Saitama, Japan). Clinicopathological factors, including age, gender, tumor size, histological subtype, tumor grade, microvascular invasion, histological tumor necrosis, C-reactive protein levels and performance status were reviewed. These factors were compared between patients with and without postoperative recurrence. Recurrence-free survival (RFS) and cause-specific survival (CSS) rates were calculated using the Kaplan-Meier method. Univariate and multivariate analyses were performed to determine independent factors predicting recurrence in patients with pT1aN0M0 RCC. The 5-year RFS and CSS rates were 97.2 and 99.1%, respectively. When clinicopathological factors were compared between patients with and without recurrence, tumor size (P=0.0390) and percentage of tumor necrosis (P<0.0001) were significantly different between groups. All patients with recurrence had primary lesions ≥3 cm. By univariate analysis, tumor size (P=0.0379) and the presence of tumor necrosis (P=0.0319) were significant predictors for recurrence; tumor necrosis was also an independent predictor for recurrence (P=0.0143). In patients with pT1b tumors ≤5 cm (recurrence rate, 16.8%; n=48), the percentage of tumor necrosis was significantly higher in patients with recurrence compared with those without (P=0.0261). This suggests that tumor necrosis may be an important predictor for recurrence in small RCCs. Although recurrence is rare in pT1a RCC, the presence of tumor necrosis may be an important predictor for recurrence. Particularly, patients presenting with pT1a RCC with histological tumor necrosis should undergo careful follow-up.

Abstract 4946: Developing anti-angiogenic therapies for human hepatocellular cancer (HCC)- studies of suntinib in the woodchuck model of hepatitis B related HCC

Abstract Introduction: Better animal models that recapitulate the liver milieu of human HCC are needed. The eastern woodchuck is an established model of human hepatitis B viral infection and spontaneously develops HCC in the context of chronic woodchuck hepatitis viral infection (WHV). The translational relevance of this model for developing anti-angiogenic therapies was evaluated using sunitinib (S), a potent oral, anti-angiogenic agent. Methods: Woodchucks were bred and inoculated at birth with titered infectious WHV pools obtained from chronic WHV carriers. By 12 months of age, the rate of chronic WHV infection was &amp;gt;60%. Carriers were followed by USG, upon developing HCC, 12 animals were randomized 1:1 to S or placebo (P) given once orally daily for 30 days. From a single treatment S PK study at 4 dose levels, n=3/group, simulations showed 12mg/kg daily was expected to be optimal for achieving steady state serum concentrations between 50- 100 ng/ml in woodchucks. Tumor size and blood flow were assessed using dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) before treatment and on day 28 using standardized protocols. At study completion or when animals were humanely euthanized, tumors and any other small nodules were fixed overnight in 10% buffered formalin. After standard processing and embedding in paraffin, 4 μm sections were prepared, deparaffinized, stained with hematoxylin-eosin (H&amp;E), studied with a Zeiss Axio Imager A1 microscope by a pathologist blinded to the treatment arm. Morphometric study of areas of necrosis were compared to the size of the tumor sections by counting view fields at medium objective magnification, and differences in percentages between the groups were compared by the Excel student's two tailed, two-sample unequal variance t-test. Results: The median age was 30 months. Four animals died during the course of the study (1P, 3 S) and were replaced. Median therapy duration was 28 days. Morphologically, portal hepatitis and pre-neoplastic lesions (foci of altered hepatocytes [FAH]) and larger areas of altered hepatocytes (AAH) were seen in all livers. Areas of tumor necrosis of varying size (20-100%) adjacent to terminal hepatic venules seen in both high grade and in rare, low grade tumor sections of S treated woodchucks were rare in P animals. The difference was highly significant (p= 6.0 E-13) between the two groups. By DCE-MRI, median tumor volume change was + 17% and -13% and necrotic tumor volume change was +22% and +190% in P and S treated animals respectively. However all MRI parameters (k trans, AUC90, median tumor volume) were not statistically significantly different between the groups. Conclusions: Angiogenic response across different stages of hepatocarcinogenesis can be studied in woodchucks. Histological necrosis in S treated woodchucks without significant DCE-MRI change and the toxicities seen with S have translational significance to human HCC. Citation Format: Alexander Pomakov, Ilia Toshkov, Sandra Buitrago, Leslie Curtin, Donald Trump, Candace Johnson, Edward Ashton, Bud Tennant, Renuka Iyer. Developing anti-angiogenic therapies for human hepatocellular cancer (HCC)- studies of suntinib in the woodchuck model of hepatitis B related HCC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4946. doi:10.1158/1538-7445.AM2014-4946

Pharmacokinetics and safety of intravitreal caspofungin.

Caspofungin exhibits potent antifungal activities against Candida and Aspergillus species. The elimination rate and retinal toxicity of caspofungin were determined in this study to assess its pharmacokinetics and safety in the treatment of fungal endophthalmitis. Intravitreal injections of 50 μg/0.1 ml of caspofungin were administered to rabbits. Levels of caspofungin in the vitreous and aqueous humors were determined using high-performance liquid chromatography (HPLC) at selected time intervals (10 min and 1, 2, 4, 8, 16, 24, and 48 h), and the half-lives were calculated. Eyes were intravitreally injected with caspofungin to obtain concentrations of 10 μg/ml, 50 μg/ml, 100 μg/ml, and 200 μg/ml. Electroretinograms were recorded 4 weeks after injections, and the injected eyes were examined histologically. The concentrations of intravitreal caspofungin at various time points exhibited an exponential decay with a half-life of 6.28 h. The mean vitreous concentration was 6.06 ± 1.76 μg/ml 1 h after intravitreal injection, and this declined to 0.47 ± 0.15 μg/ml at 24 h. The mean aqueous concentration showed undetectable levels at all time points. There were no statistical differences in scotopic a-wave and b-wave responses between control eyes and caspofungin-injected eyes. No focal necrosis or other abnormality in retinal histology was observed. Intravitreal caspofungin injection may be considered to be an alternative treatment for fungal endophthalmitis based on its antifungal activity, lower retinal toxicity, and lower elimination rate in the vitreous. More clinical data are needed to determine its potential role as primary therapy for fungal endophthalmitis.

Conditional knockout of proximal tubule mitofusin 2 accelerates recovery and improves survival after renal ischemia.

Proximal tubule (PT) cells are critical targets of acute ischemic injury. Elimination of the mitochondrial fusion protein mitofusin 2 (Mfn2) sensitizes PT cells to apoptosis in vitro. However, the role of PT Mfn2 in ischemic AKI in vivo is unknown. To test its role, we evaluated the effects of conditional KO of PT Mfn2 (cKO-PT-Mfn2) on animal survival after transient bilateral renal ischemia associated with severe AKI. Forty-eight hours after ischemia, 28% of control mice survived compared with 86% of cKO-PT-Mfn2 animals (P<0.001 versus control). Although no significant differences in histologic injury score, apoptosis, or necrosis were detected between genotypes, cKO-PT-Mfn2 kidneys exhibited a 3.5-fold increase in cell proliferation restricted to the intrarenal region with Mfn2 deletion. To identify the signals responsible for increased proliferation, primary PT cells with Mfn2 deficiency were subjected to stress by ATP depletion in vitro. Compared with normal Mfn2 expression, Mfn2 deficiency significantly increased PT cell proliferation and persistently activated extracellular signal-regulated kinase 1/2 (ERK1/2) during recovery from stress. Furthermore, stress and Mfn2 deficiency decreased the interaction between Mfn2 and Ras detected by immunoprecipitation, and purified Mfn2 dose-dependently decreased Ras activity in a cell-free assay. Ischemia in vivo also reduced the Mfn2-RAS interaction and increased both RAS and p-ERK1/2 activity in the renal cortical homogenates of cKO-PT-Mfn2 mice. Our results suggest that, in contrast to its proapoptotic effects in vitro, selective PT Mfn2 deficiency accelerates recovery of renal function and enhances animal survival after ischemic AKI in vivo, partly by increasing Ras-ERK-mediated cell proliferation.