Development of non-invasive flexible directional microwave ablation for central lung cancer: a simulation study.

Transbronchial microwave ablation (MWA) with flexible antennas has gradually become an attractive alternative to percutaneous MWA for lung cancer due to its characteristic of non-invasiveness. However, flexible antennas for the precision ablation of lung tumors that are adjacent to critical bronchial structures are still not available. In this study, a non-invasive flexible directional (FD) antenna for early stage central lung tumors surrounding the bronchia was proposed. A comprehensive numerical MWA model with the FD antenna was developed in a real human-sized left lung model. The structure of the antenna and the treatment protocol were optimized by a generic algorithm for the precision ablation of two cases of early stage central lung cancer (i.e. spherical-like and ellipsoidal tumors). The electromagnetic efficiency of the optimized antenna was also improved by implementing an optimizedπ-matching network for impedance matching. The results indicate that the electromagnetic energy of MWA can be restricted to a particular area for precision ablation of specific lung tumors using the FD antenna. This study contributes to the field of lung cancer management with MWA.

Optimal design of aperiodic tri-slot antennas for the conformal ablation of liver tumors using an experimentally validated MWA computer model.

OBJECTIVE: This study aims to demonstrate that the conformal microwave ablation (MWA) of liver tumors could be attained by optimizing the structure of an aperiodic tri-slot coaxial antenna, its insertion depth, and input power. METHODS: A computational MWA model with an aperiodic tri-slot coaxial antenna operating at the frequency of 2.45 GHz was built and validated by both an ex vivo and a pilot in vivo experiment with porcine healthy livers. The validated in vivo computational MWA model implemented with a liver tumor was then used as a testbed to investigate the conformal ablation of liver tumors. Five liver tumors in different sizes and shapes were investigated. A genetic algorithm optimization method (NSGA-II) was used to optimize the structure of antenna, insertion depth of antenna, and microwave antenna input power for the conformal ablation of liver tumors. RESULTS: The validation results showed that a good agreement in both the spatiotemporal temperature distribution and ablation zone was found between the computer model and the ex vivo experiments at both 45 W, 5 min and 60 W, 3 min treatments and the in vivo experiment at 45 W, 5 min treatment. The optimized simulation results confirmed that five cases of liver tumors in different sizes and shapes can be conformally ablated by optimizing the aperiodic tri-slot coaxial antenna, antenna insertion depth, and microwave antenna input power. CONCLUSION: This paper demonstrates that the aperiodic tri-slot coaxial antenna can be optimized with the insertion depth and input power for the conformal ablation of liver tumors, regardless the size and shape of liver tumors.

Preparation and Use of Decellularized Extracellular Matrix for Tissue Engineering.

The multidisciplinary fields of tissue engineering and regenerative medicine have the potential to revolutionize the practise of medicine through the abilities to repair, regenerate, or replace tissues and organs with functional engineered constructs. To this end, tissue engineering combines scaffolding materials with cells and biologically active molecules into constructs with the appropriate structures and properties for tissue/organ regeneration, where scaffolding materials and biomolecules are the keys to mimic the native extracellular matrix (ECM). For this, one emerging way is to decellularize the native ECM into the materials suitable for, directly or in combination with other materials, creating functional constructs. Over the past decade, decellularized ECM (or dECM) has greatly facilitated the advance of tissue engineering and regenerative medicine, while being challenged in many ways. This article reviews the recent development of dECM for tissue engineering and regenerative medicine, with a focus on the preparation of dECM along with its influence on cell culture, the modification of dECM for use as a scaffolding material, and the novel techniques and emerging trends in processing dECM into functional constructs. We highlight the success of dECM and constructs in the in vitro, in vivo, and clinical applications and further identify the key issues and challenges involved, along with a discussion of future research directions.

Radiofrequency ablation for liver tumors abutting complex blood vessel structures: treatment protocol optimization using response surface method and computer modeling.

OBJECTIVE: To achieve a result of a large tumor ablation volume with minimal thermal damage to the surrounding blood vessels by designing a few clinically-adjustable operating parameters in radiofrequency ablation (RFA) for liver tumors abutting complex vascular structures. METHODS: Response surface method (RSM) was employed to correlate the ablated tumor volume (Ra) and thermal damage to blood vessels (Dt) based on RFA operating parameters: ablation time, electrode position, and insertion angle. A coupled electric-thermal-fluid RFA computer model was created as the testbed for RSM to simulate RFA process. Then, an optimal RFA protocol for the two conflicting goals, namely (1) large tumor ablation and (2) small thermal damage to the surrounding blood vessels, has been achieved under a specific ablation environment. RESULTS: Linear regression analysis confirmed that the RFA protocol significantly affected Ra and Dt (the adjusted coefficient of determination Radj2 = 93.61% and 95.03%, respectively). For a proposed liver tumor scenario (liver tumor with a dimension of 4×3×2.9 cm3 abutting a complex vascular structure), an optimized RFA protocol was found based on the regression results in RSM. Compared with a reference RFA protocol, in which the electrode was centered in the tumor with a 12-min ablation time, the optimized RFA protocol has increased Ra  from 98.1% to 99.6% and decreased Dt from 4.1% to 0.4%, achieving nearly the complete ablation of proposed liver tumor and ignorable thermal damages to vessels. CONCLUSION: This work showed that it is possible to design a few clinically-adjustable operating parameters of RFA for achieving a large tumor ablation volume while minimizing thermal damage to the surrounding blood vessels.

Lethal Electric Field Thresholds for Cerebral Cells With Irreversible Electroporation and H-FIRE Protocols: An In Vitro Three-Dimensional Cell Model Study.

The lethal electric field (LEF) thresholds for three typical cerebral cells, including a malignant glioblastoma (GBM) cell line and two cell lines from the healthy blood-brain barrier (BBB), treated by irreversible electroporation (IRE) or high-frequency irreversible electroporation (H-FIRE) protocols were investigated in an in vitro three-dimensional (3D) cell model. A conventional IRE protocol (90 pulses, 1 Hz, and 100-µs pulse duration) and three novel H-FIRE protocols (1-3-1, 0.5-1-0.5, and 1-1-1) were used to treat the cerebral cells in both 3D single-cell and two-cell models. The electrical conductivity of the 3D cell model under different electric field strengths were characterized with the method of electrochemical impedance spectroscopy (EIS). Based on EIS, a numerical electrothermal model of electroporation was built for the determination of the LEF threshold with different protocols and temperature monitoring. Cell viability was assessed by fluorescence staining 6 h after the treatment. The results showed no thermal lethal effect on cells when these protocols were used. The LEF threshold for GBM cells was significantly lower than that of the healthy BBB cells. These results suggest the possibility of selective ablation of human cerebral GBM by IRE and H-FIRE treatments with no injury or reversible injury to healthy cells, and the potential use of IRE or H-FIRE for transient disruption of the BBB to allow chemotherapy to reach the tumor.

Modeling and ex vivo experimental validation of liver tissue carbonization with laser ablation.

OBJECTIVE: The purpose of this study was to model the process of liver tissue carbonization with laser ablation (LA). METHODS: A dynamic heat source model was proposed and combined with the light distribution model as well as bioheat transfer model to predict the development of tissue carbonization with laser ablation (LA) using an ex vivo porcine liver tissue model. An ex vivo laser ablation experiment with porcine liver tissues using a custom-made 1064 nm bare fiber was then used to verify the simulation results at 3, 5, and 7 W laser administrations for 5 min. The spatiotemporal temperature distribution was monitored by measuring the temperature changes at three points close the fiber during LA. Both the experiment and simulation of the temperature, tissue carbonization zone, and ablation zone were then compared. RESULTS: Four stages were recognized in the development of liver tissue carbonization during LA. The growth of the carbonization zone along the fiber axial and radial directions were different in the four stages. The carbonization zone along the fiber axial direction (L2) grew in the four stages with a sharp increase in the initial period and a minor increase in Stage 4. However, the change in the carbonization zone along the fiber radial direction (D2) increased dramatically (Stage 1) to a long-time plateau (Stages 2 and 3) followed by a slow growth in Stage 4. An acceptable agreement between the computer simulation and ex vivo experiment in the temperature changes at the three points was found at all three testing laser administrations. A similar result was also obtained for the dimensions of coagulation zone and ablation zone between the computer simulation and ex vivo experiment (carbonization zone: 2.99± 0.10 vs. 2.78 mm2, 67.39± 0.09 vs. 63.53 mm2, and 90.53± 0.11 vs. 85.15 mm2; ablation zone: 68.95± 0.28 vs. 65.29 mm2, 182.11± 0.24 vs. 213.81 mm2, and 244.80± 0.06 vs. 251.79 mm2 at 3, 5, and 7 W, respectively). CONCLUSION: This study demonstrates that the proposed dynamic heat source model combined with the light distribution model as well as bioheat transfer model can predict the development of liver tissue carbonization with an acceptable accuracy. This study contributes to an improved understanding of the LA process in the treatment of liver tumors.

Electroporation-Based Therapy for Brain Tumors: A Review.

Electroporation-based therapy (EBT), as a high-voltage-pulse technology has been prevalent with favorable clinical outcomes in the treatment of various solid tumors. This review paper aims to promote the clinical translation of EBT for brain tumors. First, we briefly introduced the mechanism of pore formation in a cell membrane activated by external electric fields using a single cell model. Then, we summarized and discussed the current in vitro and in vivo preclinical studies, in terms of (1) the safety and effectiveness of EBT for brain tumors in animal models, and (2) the blood-brain barrier (BBB) disruption induced by EBT. Two therapeutic effects could be achieved in EBT for brain tumors simultaneously, i.e., the tumor ablation induced by irreversible electroporation (IRE) and transient BBB disruption induced by reversible electroporation (RE). The BBB disruption could potentially improve the uptake of antitumor drugs thereby enhancing brain tumor treatment. The challenges that hinder the application of EBT in the treatment of human brain tumors are discussed in the review paper as well.

An in vivo study of a custom-made high-frequency irreversible electroporation generator on different tissues for clinically relevant ablation zones.

PURPOSE: To examine the ablation zone, muscle contractions, and temperature increases in both rabbit liver and kidney models in vivo for a custom-made high-frequency irreversible electroporation (H-FIRE) generator. MATERIALS AND METHODS: A total of 18 New Zealand white rabbits were used to investigate five H-FIRE protocols (n = 3 for each protocol) and an IRE protocol (n = 3) for the performance of the designed H-FIRE device in both liver and kidney tissues. The ablation zone was determined by using histological analysis 72 h after treatment. The extent of muscle contractions and temperature change during the application of pulse energy were measured by a commercial accelerometer attached to animals and fiber optic temperature probe inserted into organs with IRE electrodes, respectively. RESULTS: All H-FIRE protocols were able to generate visible ablation zones without muscle contractions, for both liver and kidney tissues. The area of ablation zone generated in H-FIRE pulse protocols (e.g., 0.3-1 µs, 2000 V, and 90-195 bursts) appears similar to that of IRE protocol (100 µs, 1000 V, and 90 pulses) in both liver and kidney tissues. No significant temperature increase was noticed except for the protocol with the highest pulse energy (e.g., 1 µs, 2000 V, and 180 bursts). CONCLUSION: Our work serves to complement the current H-FIRE pulse waveforms, which can be optimized to significantly improve the quality of ablation zone in terms of precision for liver and kidney tumors in clinical setting.

A review of antenna designs for percutaneous microwave ablation.

Microwave (MW) antenna is a key element in microwave ablation (MWA) treatments as the means that energy is delivered in a focused manner to the tumor and its surrounding area. The energy delivered results in a rise in temperature to a lethal level, resulting in cell death in the ablation zone. The delivery of energy and hence the success of MWA is closely dependent on the structure of the antennas. Therefore, three design criteria, such as expected ablation zone pattern, efficiency of energy delivery, and minimization of the diameter of the antennas have been the focus along the evolution of the MW antenna. To further improve the performance of MWA in the treatment of various tumors through inventing novel antennas, this article reviews the state-of-the-art and summarizes the development of MW antenna designs regarding the three design criteria.

Irreversible Electroporation Enhanced by Radiofrequency Ablation: An In Vitro and Computational Study in a 3D Liver Tumor Model.

In the present study, we used a computational and experimental study in a 3D liver tumor model (LTM) to explore the tumor ablation enhancement of irreversible electroporation (IRE) by pre-heating with radiofrequency ablation (RFA) and elucidate the mechanism whereby this enhancement occurs. Three ablation protocols, including IRE alone, RFA45 → IRE (with the pre-heating temperature of 45 °C), and RFA60 → IRE (with the pre-heating temperature of 60 °C) were investigated. Both the thermal conductivity and electrical conductivity of the 3D LTM were characterized with the change in the pre-heating temperature. The results showed, compared to IRE alone, a significant increase in the tumor ablation volume (19.59 [Formula: see text] 0.61 vs. 15.29 ± 0.61 mm3, p = 0.002 and 22.87 [Formula: see text] 0.35 vs. 15.29 ± 0.61 mm3, p < 0.001) was observed with both RFA45 → IRE and RFA60 → IRE, leading to a decrease in lethal electric filed strength (8 and 17%, correspondingly). The mechanism can be attributed to the change of cell microenvironment by pre-heating and/or a synergistic effect of RFA and IRE. The proposed enhancing method might contribute to the improvement of interventional oncology in the treatment of large tumors close to critical organs (e.g., large blood vessels and bile ducts).

What Information Are Patients Receiving from the Internet about the Operative and Nonoperative Management of Acute Appendicitis?

INTRODUCTION: Recent studies suggest that nonoperative management of appendicitis (NOMA) may be a reasonable option for managing uncomplicated acute appendicitis. We examined the Internet to see if patients are likely to find the information they need to make an informed decision between the 2 options. METHODS: A list of 29 search terms was established by a focus group and then entered into Google, resulting in 49 unique webpages, each reviewed by 3 reviewers. Consensus was obtained for bias (surgery, NOMA, or balanced), webpage type, JAMA score, reading grade, and DISCERN score, a measure of quality of written information for patients. RESULTS: Thirty of the 49 websites (61%) favored surgery, while 13 (27%) favored NOMA, and 6 sites (12%) provided balanced information. Twelve of 49 sites (24%) did not list NOMA as an option. The majority of patient-directed (11/12 = 92%) and physician-directed (7/9 = 78%) webpages favored surgery, whereas academic webpages presented a more balanced distribution. Academic and physician-directed webpages ranked higher than commercial and news webpages (median ranks 3 and 4 vs. 7.5 and 8). Only 8/49 sites (16%) mentioned that the presence of a fecalith predicts the failure of NOMA. Reading grades were almost all well above the recommended grade 8 level. CONCLUSION: Most of the webpages available on the Internet do not provide enough information, nor are they sufficiently understandable to allow most patients to make an informed decision about the current options for the management of acute appendicitis.

Protection of the transplant kidney during cold perfusion with doxycycline: proteomic analysis in a rat model.

BACKGROUND: It has been previously shown that doxycycline (Doxy) protects the kidney from preservation injury by inhibition of matrix metalloproteinase. However, the precise molecular mechanism involved in this protection from injury is not known. We used a pharmaco-proteomics approach to identify potential molecular targets associated with kidney preservation injury. METHODS: Rat kidneys were cold perfused with or without doxycycline (Doxy) for 22 h. Kidneys perfusates were analyzed for the presence of injury markers such as lactate dehydrogenase (LDH), and neutrophil-gelatinase associated lipocalin (NGAL). Proteins extracted from kidney tissue were analyzed by 2-dimensional gel electrophoresis. Proteins of interest were identified by mass spectrometry. RESULTS: Triosephosphate isomerase, PGM, dihydropteridine reductase-2, pyridine nucleotide-disulfide oxidoreductase, phosphotriesterase-related protein, and aminoacylase-1A were not affected by cold perfusion. Perfusion with Doxy increased their levels. N(G),N(G)-dimethylarginine dimethylaminohydrolase and phosphoglycerate kinase 1 were decreased after cold perfusion. Perfusion with Doxy led to an increase in their levels. CONCLUSIONS: This study revealed specific metabolic enzymes involved in preservation injury and in the mechanism whereby Doxy protects the kidney against injury during cold perfusion.

A Novel Method to Increase Tumor Ablation Zones With RFA by Injecting the Cationic Polymer Solution to Tissues: In Vivo and Computational Studies.

OBJECTIVE: This study aims to examine, for the first time, the introduction of cationic polymer solutions to improve radiofrequency ablation (RFA) in terms of a potentially enlarged ablation zone. METHODS: By using in vivo and computational RFA studies, two cationic polymers, Chitooligosaccharides (COS) and carboxymethyl chitosan (CMC), diluted in deionized water, were injected into tissues separately surrounding the RF bipolar electrode prior to power application. A total of 9 rabbits were used to 1) measure the increase in electrical conductivity of tissues injected with the cationic polymer solutions, and 2) explore the enhancement of the ablation performance in RFA trials. A computer model of RFA comprising a model of the solution diffusion with an RF thermal ablation model was also built, validated by the in vivo experiment, to quantitatively study the effect of cationic polymer solutions on ablation performances. RESULTS: Compared to the control group, the electrical conductivity of rabbit liver tissues was increased by 42.20% (0.282 ± 0.006 vs. 0.401 ± 0.048 S/m, P = 0.001) and 43.97% (0.282 ± 0.006 vs. 0.406 ± 0.042 S/m, P = 0.001) by injecting the COS and CMC solution at the concentration of 100 mg/mL into the tissues, denoted COSDW100 and CMCDW100, respectively. Consequently, the in vivo experiments show that the ablation zone was enlarged by 95% (47.6 ± 6.3 vs. 92.6 ± 11.5 mm2, P < 0.001) and 87% (47.6± 6.3 vs. 88.8 ± 9.6 mm2, P < 0.001) by COSDW100 and CMCDW100, respectively. The computer simulation shows that the ablation zone was enlarged by 71% (51.9 vs. 88.7 mm2) and 63% (51.9 vs. 84.7 mm2) by COSDW100 and CMCDW100, respectively. CONCLUSION: The injection of the cationic solution can greatly improve the performance of RFA treatment in terms of enlarging the ablation zone, which is due to the increase in the electrical conductivity of liver tissues surrounding the RF electrode. SIGNIFICANCE: This study contributes to the improvement of RFA in the treatment of large tumors.

Chemical Enhancement of Irreversible Electroporation: A Review and Future Suggestions.

Irreversible electroporation has raised great interest in the past decade as a means of destroying cancers in a way that does not involve heat. Irreversible electroporation is a novel ablation technology that uses short high-voltage electrical pulses to enhance the permeability of tumor cell membranes and generate irreversible nano-sized structural defects or pores, thus leading to cell death. Irreversible electroporation has many advantages over thermal therapies due to its nonthermal mechanism: (1) reduced risk of injury to surrounding organs and (2) no "heat-sink" effect due to nearby blood vessels. However, so far, it has been difficult for irreversible electroporation to completely ablate large tumors (eg, >3 cm in diameter). In order to overcome this problem, many preclinical and clinical studies have been performed to improve the efficacy of IRE in the treatment of large size of tumors through a chemical perspective. Due to the distribution of electric field, irreversible electroporation region, reversible electroporation region, and intact region can be found in the treatment of irreversible electroporation. Thus, 2 types of chemical enhancements of irreversible electroporation were discussed in the article, such as the reversible electroporation region enhanced and the irreversible electroporation region enhanced. Specifically, the state-of-the-art results regarding the following approaches that have the potential to be used in the enhancement of irreversible electroporation were systematically reviewed in the article, including (1) combination with cytotoxic drugs, (2) calcium electroporation, (3) modification of cell membrane, and (4) modification of the tumor cell microenvironment. In the end, we concluded with 4 issues that should be addressed in the future for improving irreversible electroporation further in a chemical way.

Pharmacological Protection of Kidney Grafts from Cold Perfusion-Induced Injury.

One of the greatest challenges facing the field of organ transplantation is the shortage of donor organs for transplantation. Renal transplantation increases quality of life and survival of patients suffering from end-stage renal disease. Although kidney transplantation has evolved greatly over the past few decades, a not insignificant amount of injury occurs to the kidney during recovery, preservation, and implantation and leads to the loss of function and loss of years of dialysis-free living for many patients. The use of kidneys from expanded criteria donors (ECD) and donation after circulatory determination of death (DCDD) has been adopted partly in response to the shortage of donor kidneys; however these kidneys are even more susceptible to ischemic injury. It has been shown that matrix metalloproteinases (MMPs) and reactive oxygen species (ROS) are involved in mechanisms of injury to the transplant kidney. There is also some evidence that inhibition of MMP activity and/or ROS production can protect the kidney from injury. We review possible pharmacological strategies for protection of kidney graft from injury during recovery, preservation, and implantation.

Direction of the Biopsy Needle in Ultrasound-Guided Renal Biopsy Impacts Specimen Adequacy and Risk of Bleeding.

INTRODUCTION: Although medical factors such as hypertension and coagulopathy have been identified that are associated with hemorrhage after renal biopsy, little is known about the role of technical factors. The purpose of our study was to examine the effects of biopsy needle direction on renal biopsy specimen adequacy and bleeding complications. METHODS: Two hundred and forty-two patients who had undergone ultrasound-guided renal biopsies were included. A printout of the ultrasound picture taken at the time of the biopsy was used to measure the biopsy angle ("angle of attack" [AOA]) and to determine if the biopsy needle was aimed at the upper or lower pole and if the medulla was targeted or avoided. RESULTS: Of the 3 groups of biopsy angle, an AOA of between 50°-70° yielded the most glomeruli per core (P = .001) and the fewest inadequate specimens (4% vs 15% for > 70°, and 9% for < 50°, P = .038). Biopsy directed at a pole vs an interpolar region resulted in fewer inadequate specimens (8% vs 23%, P = .005), while biopsies that were medulla-avoiding resulted in fewer inadequate specimens (5% vs 16%, P = .004) and markedly reduced bleeding complications (12% vs 46%, P < .001) compared to biopsies where the medulla was entered. DISCUSSION: An AOA of approximately 60°, aiming at the poles, and avoiding the medulla were each associated with fewer inadequate biopsies and bleeding complications. While biopsy of the medulla is necessary for some diagnoses, the increased bleeding risk emphasizes the need for communication between nephrologist, pathologist, and radiologist.

Tumor Ablation Enhancement by Combining Radiofrequency Ablation and Irreversible Electroporation: An In Vitro 3D Tumor Study.

We hypothesized and demonstrated for the first time that significant tumor ablation enhancement can be achieved by combining radiofrequency ablation (RFA) and irreversible electroporation (IRE) using a 3D cervical cancer cell model. Three RFA (43, 50, and 60 °C for 2 min) and IRE protocols (350, 700, and 1050 V/cm) were used to study the combining effect in the 3D tumor cell model. The in vitro experiment showed that both RFA enhanced IRE and IRE enhanced RFA can lead to a significant increase in the size of the ablation zone compared to IRE and RFA alone. It was also noted that the sequence of applying ablation energy (RFA â†’ RE or IRE â†’ RFA) affected the efficacy of tumor ablation enhancement. The electrical conductivity of 3D tumor was found to be increased after preliminary RFA or IRE treatment. This increase in tumor conductivity may explain the enhancement of tumor ablation. Another explanation might be that there is repeat injury to the transitional zone of the first treatment by the second one. The promising results achieved in the study can provide us useful clues about the treatment of large tumors abutting large vessels or bile ducts.

Transplant renal vein thrombosis in a recipient with aberrant venous anatomy.

Renal vein thrombosis in a transplanted kidney is an uncommon but critical complication that can result in graft loss if management is delayed. A 31-year-old male with known atresia of the inferior vena cava who received a deceased donor renal transplant 7 years previously presented to hospital with severe graft site pain and a week of nausea, vomiting, and chills. Serum creatinine was markedly elevated from baseline. Sonographic examination revealed external iliac vein thrombosis with extension of the thrombus into the transplant renal vein. Urgent angiographic administration of tissue plasminogen activator and suction thrombectomy was performed, then followed by heparin and clopidogrel post procedure. Within 24 h, his serum creatinine improved, and within 2 weeks returned to his baseline. He was started on lifelong warfarin anti-coagulation to reduce the risk of rethrombosis secondary to his uncorrectable aberrant venous anatomy. Due to the turbulent and sometimes reversed flow in the major veins, lifelong anticoagulation should be strongly considered for such transplant patients with recipient aberrancy of the large veins.

Development of a statistical model for cervical cancer cell death with irreversible electroporation in vitro.

PURPOSE: The aim of this study was to develop a statistical model for cell death by irreversible electroporation (IRE) and to show that the statistic model is more accurate than the electric field threshold model in the literature using cervical cancer cells in vitro. METHODS: HeLa cell line was cultured and treated with different IRE protocols in order to obtain data for modeling the statistical relationship between the cell death and pulse-setting parameters. In total, 340 in vitro experiments were performed with a commercial IRE pulse system, including a pulse generator and an electric cuvette. Trypan blue staining technique was used to evaluate cell death after 4 hours of incubation following IRE treatment. Peleg-Fermi model was used in the study to build the statistical relationship using the cell viability data obtained from the in vitro experiments. A finite element model of IRE for the electric field distribution was also built. Comparison of ablation zones between the statistical model and electric threshold model (drawn from the finite element model) was used to show the accuracy of the proposed statistical model in the description of the ablation zone and its applicability in different pulse-setting parameters. RESULTS: The statistical models describing the relationships between HeLa cell death and pulse length and the number of pulses, respectively, were built. The values of the curve fitting parameters were obtained using the Peleg-Fermi model for the treatment of cervical cancer with IRE. The difference in the ablation zone between the statistical model and the electric threshold model was also illustrated to show the accuracy of the proposed statistical model in the representation of ablation zone in IRE. CONCLUSIONS: This study concluded that: (1) the proposed statistical model accurately described the ablation zone of IRE with cervical cancer cells, and was more accurate compared with the electric field model; (2) the proposed statistical model was able to estimate the value of electric field threshold for the computer simulation of IRE in the treatment of cervical cancer; and (3) the proposed statistical model was able to express the change in ablation zone with the change in pulse-setting parameters.

Proteomic Analysis of Perfusate from Machine Cold Perfusion of Transplant Kidneys: Insights Into Protection from Injury.

BACKGROUND Machine cold perfusion is beneficial to the preservation of kidneys for transplantation. At the end of preservation, the perfusion solution contains many proteins. Using a proteomics approach, we searched for useful biomarkers and potential therapeutic targets in the perfusate. Our program is unique in that all transplant kidneys (even living donor kidneys, LKD) are placed on machine cold perfusion prior to transplantation. MATERIAL AND METHODS Perfusates from donation after neurological and circulatory determination of death (DNDD and DCDD respectively) and LKD were collected (n=41) and analyzed for LDH, neutrophil gelatinase-associated lipocalin (NGAL), and matrix metalloproteinase-2 (MMP-2) as markers of injury. Perfusate from each kidney was subjected to 2-dimensional gel electrophoresis, then analyzed using software to identify those spots which are significantly different between the 3 groups. Mass spectrometry was used to identify the proteins and their identity was confirmed with Western blot. RESULTS The highest levels of MMP-2, LDH, and NGAL were seen for the DCDD kidneys, followed by the DNDD kidneys and then LDK. Peroxiredoxin-2, NGAL, and alpha-1-antitrypsin were identified as significantly different between the different types of donor kidneys, and their role and possible therapeutic strategies are discussed. Collagen fragments, albumin, and immunoglobulin were also identified as possible byproducts of the injury and may be useful is assessing the degree of injury. CONCLUSIONS Comparison of the perfusates from the different types of kidneys has allowed us to identify proteins that will be useful in future research into reducing injury in transplant kidneys.