Non-thermal treatment of endometriosis

Abstract

A device, system and methods for the non-thermal treatment of endometriosis.

Description

Arnold & Porter Ref. P35370WO00 NON-THERMAL TREATMENT OF ENDOMETRIOSIS CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No.63 / 586,190, filed September 28, 2023, and U.S. Provisional Application No.63 / 697,881, filed September 23, 2024, both of which are incorporated by reference herein in their entireties. FIELD OF THE INVENTION

[0002] The present disclosure relates to non-thermal treatments of endometriosis and more particularly to methods and devices for electric field treatment and removal of endometrial lesions in a subject in need thereof. BACKGROUND OF THE INVENTION

[0003] Endometriosis is a condition caused by the growth of endometrial cells outside of their normal environment, the uterus. For this condition, endometrial cells, which typically thicken and break-down in the uterus during a female’s menstrual cycle, are found in other tissues, for example in the ovaries, fallopian tubes and pelvis. These cells, although not in the uterus, continue to function (thicken and then break-down) and respond to hormonal stimulus as if they are within the uterus, resulting in pain and other symptoms.

[0004] It is estimated that about 3 million new cases of endometriosis occur each year, particularly in women between the ages of 25 and 40, women who have a family history of endometriosis without children, and women who suffer unusual menstrual cycles.

[0005] Endometriosis can cause pelvic cramping and pain, lower back pain, abdominal pain, excessive menstrual bleeding, fatigue, possible problems with fertility, and has been associated with a higher likelihood of developing ovarian cancer. These symptoms of the condition can be fairly mild in nature, or be more severe in nature, resulting in treatment options that range from over-the-counter pain medications to surgical intervention.

[0006] Conventional treatment options, as noted above, depend on the severity of the endometriosis and the patient’s symptoms. A number of non-invasive treatments depend on interrupting the natural hormonal cycle of the effected woman, including hormonal therapies, GnRH medications, oral contraceptives, and the like. These treatments limit the natural process of ovulation and thereby prevent the break-down of the endometrial tissue outside the uterus (the source of much of the pain). In more severe cases, a surgical intervention is necessitated for removal of endometrial tissue, particularly where the tissue has caused a significant lesion. In theArnold & Porter Ref. P35370WO00 most severe cases, hysterectomy with or without bilateral salpingo-oophorectomy is called for, thereby removing some, if not all of the offending tissue. More typically, endometrial lesions are removed through the use of laser ablation, where the offending tissue is heated to removal and the surrounding tissue and blood vessels sealed.

[0007] These thermal ablation surgical solutions often result in heat induced damage to the surrounding healthy tissue, which then leaves scarring / fibrosis where the lesion has been removed. These heat induced damages can have a negative impact on the subject’s fertility, particularly if the treatment involves the ovaries.

[0008] The present disclosure provides alternative devices and treatment methods to treat endometrial lesion with reduced damage. SUMMARY OF THE INVENTION

[0009] The present disclosure provides for, and includes, a device for non-thermal treatment of one or more lesions in a subject in need thereof comprising: a generator 110 comprising a first output connector and a second output connector; an applicator 200 comprising a first electrode 210 and a second electrode 220; a dispersive pad 300; and a switch 410 in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator.

[0010] The present disclosure also provides for, and includes, a device for an application of a non-thermal treatment dose comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a first electrode and a second electrode; a first switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply a therapeutic electric field of a first mode through the applicator and a second position to apply a therapeutic electric field of a second mode through the applicator; a dispersive pad in an electrical connection with the second output connector; a sensor configured to receive a first signal before the application of a non-thermal treatment dose and a second signal after the application of the non-thermal treatment dose; a processor electronically coupled to the switch and the sensor, a non-transitory computer readable medium electronically coupled to the processor, wherein the non-transitory computer readable medium comprises instructions stored thereon that, when executed on the processor, perform the steps of receiving the first signal from the sensor and the second signal from the sensor; and determining a difference between the first signal and the second signal, comparing the difference against a predetermined threshold; and instructing the application to end if the difference exceeds the predetermined threshold.Arnold & Porter Ref. P35370WO00

[0011] The present disclosure also provides for and includes a device for an application of a non-thermal treatment dose comprising: a generator 110 comprising a first output connector and a second output connector; an applicator 200 comprising a first electrode 210 in a first electrical connection with the first output and a second electrode 220 in a second electrical connection with the second output; a switch in a first electrical connection with the first output connector and in a second electrical connection with the first electrode, wherein the switch in a first position is configured to apply a therapeutic electric field through the first electrode, and wherein the switch in a second position is configured to disrupt the first electrical connection to the first output connector and the second electrical connection with the first electrode; a sensor configured to receive a first signal before the application of a non-thermal treatment dose and a second signal after the application of the non-thermal treatment dose; a processor electronically coupled to the switch and the sensor; a non-transitory computer readable medium electronically coupled to the processor, wherein the non-transitory computer readable medium comprises instructions stored thereon that, when executed on the processor, perform the steps of receiving the first signal from the sensor and the second signal from the sensor; and determining a difference between the first signal and the second signal, comparing the difference against a predetermined threshold; and instructing the switch to configure to the second position. In an aspect, applicator 200 is an articulating tip applicator 201.

[0012] The present disclosure also provides for, and includes, a device for non-thermal treatment comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a first electrode and a second electrode; a dispersive pad; and a first switch 410 in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator; a second switch 420 upstream of the first switch and in a first electrical connection with the first output connector and in a second electrical connection with the first electrode, wherein the second switch is configured to disrupt the first electrical connection to the first output connector and the second electrical connection with the first electrode; a sensor configured to recognize a signal; a processor electronically coupled to the switch and configured to receive a second signal from the sensor; and a non-transitory computer readable medium electronically coupled to the processor, wherein the non-transitory computer readable medium comprises instructions stored thereon activated, after the receipt of the signal, to perform the steps of disconnecting the electrical connection between the first electrical connection and the first output.Arnold & Porter Ref. P35370WO00

[0013] Moreover, the present disclosure also provides for, and includes, a method of non-thermal treatment of a lesion in a subject in need thereof comprising: identifying the location, size and volume of the lesion in the subject; determining an electric field useful in the treatment of the identified lesion; and applying the electric field to the lesion for an appropriate time and appropriate voltage using a device comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a first electrode and a second electrode; a dispersive pad; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator.

[0014] A further aspect of the present disclosure is a method for non-thermal treatment of a lesion in a subject in need thereof comprising: determining an electric field useful in the treatment of the lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the lesion with a device comprising: a generator comprising a first output and a second output; an applicator comprising a first electrode and a second electrode; a dispersive pad; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply the electric field of the first mode through the applicator and a second position to apply the electric field of the second mode through the applicator, wherein the electric field is within a treatment area in the subject in need thereof.

[0015] An aspect of the present disclosure provides for, and includes, a method of causing controlled cell necrosis in an endometrium of a subject in need thereof comprising: determining an electric field useful in the treatment of an endometrial lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the endometrial lesion with a device comprising: a generator comprising a first output and a second output, an applicator comprising a first electrode and a second electrode; a dispersive pad; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply the electric field of the first mode through the applicator and a second position to apply the electric field of the second mode through the applicator, wherein the electric field of the first mode or the electric field of the second mode is within a treatment area of the subject in need thereof and causes cell necrosis within the treatment area.

[0016] A further aspect of the present disclosure provides for, and includes, an articulating tip applicator 202 comprising: a shaft comprising: a distal end comprising a first electrode 210 comprises a first stainless steel rod 212, a distal end 213, and a proximal end 214; a second electrode 220 comprises a second stainless steel tube 222, a distal end 223, and a proximal endArnold & Porter Ref. P35370WO00 224, and an insulating layer; a flexible section; and a proximal end comprising mechanical levers connected to the flexible section with cables, wherein the flexible section is situated between the proximal end and the distal end, and wherein the first electrode 210 and the second electrode 220 are concentric, and the proximal end 214 of the first electrode and the distal end 223 of the second electrode are separated by a distance in length 230w. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Some aspects of the disclosure are herein described, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and are for purposes of illustrative discussion of embodiments of the disclosure. In this regard, the description, taken with the drawings, makes apparent to those skilled in the art how aspects of the disclosure may be practiced.

[0018] FIG.1A illustrates an exemplary embodiment of a non-thermal treatment system of the present disclosure;

[0019] FIG.1B illustrates an exemplary embodiment of a non-thermal treatment system of the present disclosure;

[0020] FIG.2A illustrates an exemplary embodiment of an applicator 200 for use in a non- thermal treatment system of the present disclosure;

[0021] FIG.2B illustrates an exemplary embodiment of a clamping probe 201 for use in a non- thermal treatment system of the present disclosure;

[0022] FIG.3A illustrates an exemplary embodiment of an articulating tip applicator 202 in a retracted position, for use in a non-thermal treatment system of the present disclosure;

[0023] FIG.3B illustrates an exemplary embodiment of articulating tip applicator 202 in extended position, for use in a non-thermal treatment system of the present disclosure;

[0024] FIG.4A illustrates an exemplary embodiment of a catheter (top) for use in a non- thermal treatment system (bottom) in bipolar mode of the present disclosure;

[0025] FIG.4B illustrates an exemplary embodiment of a catheter (top) for use in a non-thermal treatment system in monopolar mode 520 of the present disclosure;

[0026] FIG.5 depicts an exemplary waveform view of a non-thermal treatment system of the present disclosure;

[0027] FIG.6 depicts another exemplary waveform view of a non-thermal treatment system of the present disclosure;

[0028] FIG.7A depict yet another exemplary waveform view of a non-thermal treatment system of the present disclosure;Arnold & Porter Ref. P35370WO00

[0029] FIG.7B depicts yet another exemplary waveform view of a non-thermal treatment system of the present disclosure;

[0030] FIG.8 depicts an exemplary embodiment of a clamping probe 201 in treatment of the present disclosure;

[0031] FIG.9 depicts an exemplary waveform and dose definition of the present disclosure;

[0032] FIGS.10A is a graph showing IRE resistance data including recorded averages of resistance at start of each dose;

[0033] FIG.10B is a graph showing changes in IRE resistance;

[0034] FIG.11 is a graph showing voltage droop data including an average voltage at start and end;

[0035] FIGS.12A illustrates exemplary embodiments of geometry used in computer simulation of the present disclosure;

[0036] FIG.12B illustrates exemplary embodiments of geometry used in computer simulation of the present disclosure;

[0037] FIG.13 is a graph showing comparison of recorded and simulated currents;

[0038] FIG.14 illustrates exemplary embodiments of simulation data at edges of treatment zone based on electric field magnitude.

[0039] FIG.15 depicts an image from a pathology report of tissue of low-dose bowel applications;

[0040] FIG.16 depicts an image from a pathology report of tissue of high-dose bowel applications;

[0041] FIG.17 depicts another pictorial view of low-dose bowel applications from pathology report identifying adjacent normal tissue;

[0042] FIG.18A is an exemplary embodiment of an experimental apparatus for determining the therapeutic signature;

[0043] FIG.18B is an exemplary result from treatment zones in a potato after 24 hours;

[0044] FIGS.19A depicts an exemplary embodiment of an applicator 200 in a non-thermal treatment system of the present disclosure;

[0045] FIG.19B depict an exemplary embodiment of treatment with a potato using articulating tip applicator 202;

[0046] FIG.20 is a graph showing magnitude of the potato before and after IRE treatments;

[0047] FIG.21 is a graph showing phase of the potato before and after IRE treatments;

[0048] FIG.22 is a graph showing linear regression plots to quantify the change;

[0049] FIG.23 is a graph showing linear regression plots to quantify the change;Arnold & Porter Ref. P35370WO00

[0050] FIG.24 is a graph showing a plot of magnitude and phase with the catheter in tap water;

[0051] FIG.25A illustrates a Cole-Cole equivalent circuit model.

[0052] FIG.25B illustrates magnitude and phase of the Impedance as reported by Zhao, Y., et al., “Ablation outcome of irreversible electroporation on potato monitored by impedance spectrum under multi-electrode system.” Biomedical Engineering Online, 126 (2018).

[0053] FIG.26 depicts an example of a process for testing a non-thermal treatment system according to one embodiment of the present disclosure;

[0054] FIGS.27A, 27B, 27C, and 27D depict applicators along with the tissues treated. (A) Large inner electrode, articulating tip applicator 202 (or Applicator 06Sep2023A); (B) Smaller inner electrode, articulating tip applicator 202 (or Applicator 08Sep2023A); (C) depicts treatment of the peritoneum; and (D) depicts treatment of the bowel (jejunum).

[0055] FIG.28 is a graph showing magnitude plots of the larger electrode applicator, Articulating tip applicator 202 on peritoneum, before (solid lines) and after (dashed lines) IRE application.

[0056] FIG.29 is a graph showing various phase plots of articulating tip applicator 202 on peritoneum, before (solid lines) and after (dashed lines) IRE application.

[0057] FIG.30 is a graph showing magnitude difference after IRE treatment of jejunum using the large electrode;

[0058] FIG.31 is a graph showing phase difference after IRE treatment of the jejunum with large electrode;

[0059] FIG.32 is a graph showing magnitude difference after IRE treatment of the jejunum with a small electrode;

[0060] FIG.33 is a graph showing phase difference after IRE treatment of the jejunum using small electrode;

[0061] FIG.34 shows an image of a jejunum treated with medium dose (apps 15 and 16) and high (apps 17 and 18) after treatment;

[0062] FIG.35 is a graph showing a plot of calculated IRE resistances with the technique shown on the inset image;

[0063] FIG.36 is a graph showing the magnitude difference before and after IRE treatment of peritoneum with a large electrode over the frequency range;

[0064] FIG.37 is a graph of the magnitude difference before and after IRE treatment for large electrode jejunum over the frequency range;

[0065] FIG.38 is a graph of the magnitude difference before and after IRE treatment of jejunum with a small electrode over the frequency range;Arnold & Porter Ref. P35370WO00

[0066] FIG.39 is a graph of plots of the phase difference before and after IRE treatment of peritoneum with the large electrode peritoneum over the frequency range;

[0067] FIG.40 depicts pictorial view of plots of the phase difference before and after IRE treatment of jejunum with the large electrode over the frequency range;

[0068] FIG.41 is a graph showing the magnitude difference before and after IRE treatment of jejunum with the small electrode jejunum over the frequency range;

[0069] FIGS.42A−42B depict images of postmortem porcine uterine horn treated with 800 V, 30 groups (A) before and (B) after treatment;

[0070] FIG.43 is a graph showing magnitude of small electrode on porcine postmortem uterine horn;

[0071] FIG.44 is a graph showing phase of a small electrode on porcine postmortem uterine horn;

[0072] FIGS.45A−45C illustrate an exemplary embodiment of an alternative non-thermal treatment system of the present disclosure shown as a three dimensional schematic of applicator 1200 (FIG.45A), a two-dimensional schematic of the side of applicator 1200 (FIG.45B), and a two-dimensional schematic of the top of applicator 1200 (FIG.45C);

[0073] FIG.46 illustrates an exemplary embodiment of computer simulations of an IRE zone created by using a non-thermal treatment system of the present disclosure shown as IRE zone created by catheter application (FIGS.46A, 46B, and 46C), a two-dimensional schematic of the top of applicator 1200 (FIG.46D), and a two-dimensional schematic of the side of applicator 1200 (FIG.46E);

[0074] FIG.47A−47D illustrates an exemplary embodiment of computer simulations of IRE zones created using a non-thermal treatment system of the present disclosure including applicator 1200;

[0075] FIG.48A−48C illustrates an exemplary embodiment of an opening 1207 of applicator 1200 for use in a non-thermal treatment system of the present disclosure;

[0076] FIGS.49A−49E illustrate an exemplary embodiment of various components of applicator 1200 for use in an alternative non-thermal treatment system of the present disclosure;

[0077] FIG.50A illustrates an exemplary embodiment of an applicator 1200 tip for use in an alternative non-thermal treatment system (bottom) in bipolar mode of the present disclosure;

[0078] FIG.50B illustrates an exemplary embodiment of an applicator 1200 tip for use in an alternative non-thermal treatment system in monopolar mode of the present disclosure;

[0079] FIG.51 depicts an exemplary embodiment of bipolar articulating tip applicator 202 and bipolar clamping probe in treatment according to the present disclosure;Arnold & Porter Ref. P35370WO00

[0080] FIG.52 is a graph showing magnitude plots of an exemplary bipolar clamping probe applicator (Clamp6) on ureter of the present disclosure;

[0081] FIG.53 is a graph showing phase plots of an exemplary bipolar clamping probe applicator (Clamp6) on ureter of the present disclosure;

[0082] FIG.54 is a graph showing a plot of calculated IRE resistances with the technique shown in the inset image;

[0083] FIG.55 depicts simulated electric field of non-thermal treatment system in a) peritoneal tissue and b) ureter;

[0084] FIG.56 is a graph showing the magnitude difference before and after IRE treatment of ureter over the frequency range;

[0085] FIG.57 is a graph showing the phase difference before and after IRE treatment of ureter over the frequency range;

[0086] FIG.58 depicts an exemplary embodiment of a bipolar clamp on post mortem porcine uterine horn of the present disclosure;

[0087] FIG.59 depicts an exemplary embodiment of a bipolar clamping probe on post mortem porcine uterine horn of the present disclosure

[0088] FIG.60 is a graph showing magnitude plots of clamping probe on post mortem porcine uterine horn of the present disclosure;

[0089] FIG.61 is a graph showing phase plots of clamping probes on post mortem porcine uterine horn of the present disclosure;

[0090] FIG.62 depicts an image of treatment of postmortem small bowel using articulating tip applicator 202;

[0091] FIGS.63−64 are various graphs showing magnitude plots before and after treatment on small bowel and linear curve fit of the present disclosure;

[0092] FIGS.65−66 are various graphs showing phase plots before and after treatment on small bowel and linear curve fit of the present disclosure;

[0093] FIGS.67−68 are various graphs showing magnitude plots before and after treatment on small bowel and linear curve fit of the present disclosure;

[0094] FIGS.69−70 are various graphs showing phase plots before and after treatment on small bowel and linear curve fit of the present disclosure;

[0095] FIG.71 depicts a comparison of slopes in linear curve fit of the present disclosure;

[0096] FIG.72 is an exemplary embodiment of an experimental setup and apparatus to record measurements before and after IRE treatment;Arnold & Porter Ref. P35370WO00

[0097] FIG.73 depicts applicators along with the tissues treated according to one embodiment of the present disclosure;

[0098] FIG.74 is a graph showing phase plots according to one embodiment of the present disclosure;

[0099] FIG.75 depicts a comparison of slopes after IRE treatment of the present disclosure;

[0100] FIG.76 depicts an image of application according to one embodiment of the present disclosure;

[0101] FIG.77 is a graph showing phase plots according to one embodiment of the present disclosure;

[0102] FIGS.78−79 are graphs showing magnitude plots according to one embodiment of the present disclosure.

[0103] FIGS.80−81 depict device and settings used for postmortem porcine small bowel experiments of the present disclosure;

[0104] FIGS.82−83 depict plots of fitted curve of the phase before and after IRE treatment of the present disclosure;

[0105] FIG.84 depicts difference in phase slope after IRE treatment of the present disclosure;

[0106] FIG.85 depicts images of lesions after treatment according to one embodiment of the present disclosure;

[0107] FIG.86A−86B is an exemplary embodiment of an experimental setup and apparatus according to one embodiment of the present disclosure;

[0108] FIG.87A−87C depicts images of distal end of three bipolar clamps of an exemplary embodiment;

[0109] FIG.88A−88C depicts images of each device clamped onto tissue according to one embodiment of the present disclosure;

[0110] FIGS.89−90 are graphs showing slopes of phase before and after using BiClamp2 device according to one embodiment of the present disclosure;

[0111] FIG.91 is a graph showing difference for each set of plots in FIGS.89-90;

[0112] FIGS.92−93 are graphs showing slopes of phase before and after using Maryland1 device according to one embodiment of the present disclosure;

[0113] FIG.94 is a graph showing difference for each set of plots in FIGS.92-93;

[0114] FIG.95A−95B depicts treatment zone of BiClamp5 and Maryland of an exemplary embodiment;

[0115] FIGS.96−97 are graphs showing slopes of phase before and after for using BiClamp2 device according to one embodiment of the present disclosure;Arnold & Porter Ref. P35370WO00

[0116] FIG.98 is a graph showing difference for each set of plots in FIGS.96-97;

[0117] FIG.99A−99C depicts treatment zones with thermal damage for Clamp2 of an exemplary embodiment;

[0118] FIG.100 depicts phase shift between IRE voltage and IRE current for clamp probe and articulating tip applicator 202 according to one embodiment of the present disclosure;

[0119] FIG.101 illustrates an exemplary embodiment of various components according to one embodiment of the present disclosure;

[0120] FIG.102 is a graph showing slope of a phase of clamped tissue before and after the IRE application;

[0121] FIG.103 illustrates difference in a fitted line of a phase slope;

[0122] FIG.104 plots difference of a phase slope fitted curve for 4 BiClamp8 applications;

[0123] FIG.105 depicts treatment zones of an exemplary embodiment for BiClamp7 device;

[0124] FIG.106 depicts treatment zones of an exemplary embodiment for BiClamp8 device;

[0125] FIGS.107 depicts a phase of the clamped tissue before and after the IRE application for according to one embodiment of the present disclosure;

[0126] FIG 108 depicts a phase of the clamped tissue before and after the IRE application for according to one embodiment of the present disclosure;

[0127] FIG 109 depicts a phase of the clamped tissue before and after the IRE application for according to one embodiment of the present disclosure;

[0128] FIG 110 depicts a phase of the clamped tissue before and after the IRE application for according to one embodiment of the present disclosure;

[0129] FIG.111 depicts treatment zones of an exemplary embodiment for BiClamp7 device; and

[0130] FIG.112 depicts treatment zones of an exemplary embodiment for BiClamp8 device.

[0131] Corresponding reference characters indicate corresponding parts throughout the several views. The examples set out herein illustrate embodiments of the present disclosure but should not be construed as limiting the scope of the present disclosure in any manner. DETAILED DESCRIPTION

[0132] Aspects in accordance with the present disclosure include devices and methods for non- thermal treatment of endometriosis. Endometriosis is a condition where endometrial tissue grows and proliferates outside its normal environment, the uterus. As such, endometriosis is characterized by endometrial tissue located, for example, in the ovaries, fallopian tubes, and tissues that line the pelvis. In extreme cases, the endometrial tissue can be found outside the woman’s pelvis, althoughArnold & Porter Ref. P35370WO00 this is fairly rare. Although not found in the uterus, the endometrial tissue is still regulated by a woman’s normal monthly hormonal cycle, ending with the breakdown of the tissue. Results of the endometrial tissue break-down outside the uterus can result in inflammation, swelling, and scarring to the surrounding and underlying tissue, all of which can result in pain, bloating, bleeding, and additional symptoms. In cases where the ovaries are involved, infertility can also result, as well as an increased likelihood of developing ovarian cancer.

[0133] Approximately 2-10% of women between the ages of 25 and 40 have endometriosis. Conventional therapeutics for these women can include analgesics and pain relievers. In more difficult cases, laparoscopy is used to thermally remove the endometrial lesion and in very difficult cases, a hysterectomy (removal of the uterus, fallopian tubes and ovaries) is performed. Where laparoscopy is the primary treatment, the removal results in destruction of the endometrial cells and matrix, with subsequent fibrosis and scarring of the damaged area. The present disclosure provides for and includes non-thermal treatment system 100 and methods of using the non-thermal treatment system and the included applicators for the removal of endometrial lesions with minimal or absence of endothermal damage. In an aspect, electrical fields of appropriate strength are used to introduce electroporation of cell membranes in treatment zones, thereby causing necrosis of the endometrial cells but leaving the intervening matrix intact for the treatment of a patient in need thereof.

[0134] Without limiting to theory, aspects of the present disclosed system and methods deliver an electric field sufficient in size and strength to cause electroporation of the treated cells, but not to result in damage to the connective tissue that surround and separate the endometrial cells. This method of delivering an electric field, in the absence of thermal ablation, causes necrosis of the endometrial cells but limits scarring, major benefits of the present disclosure.

[0135] Referring to FIG.1A, a non-thermal treatment system 100 of the present disclosure is shown. The non-thermal treatment system 100 comprising a device for non-thermal treatment of one or more lesions of a patient. In an aspect of the present disclosure, a non-thermal treatment system 100 comprises a generator 110, an applicator 200, a dispersive pad 300, and a switch 400 in electrical connection with the generator 110. In an aspect of the present disclosure, the dispersive pad 300 is detachable from the non-thermal treatment system 100. In an aspect of the present disclosure, a non-thermal treatment system 100 comprising a generator 110, an applicator 200, a dispersive pad 300, a first switch 410 in electrical connection with the generator 110, and a second switch 420 upstream of switch 410.

[0136] In an aspect of the present disclosure, the non-thermal treatment system comprises one or more switches 400. In an aspect, the non-thermal treatment system comprises a first switch 410 configured to change between a first position to apply an electric field of a first mode through theArnold & Porter Ref. P35370WO00 applicator and a second position to apply an electric field of a second mode through the applicator. In an aspect, switch 410 is a mechanical switch. In an aspect, switch 410 is an electronic switch. In an aspect, switch 410 is a latch switch. In an aspect, switch 410 is a momentary switch. In a further aspect, switch 410 is a double pole double throw switch, a single pole double throw switch, or a changeover switch. In an aspect of the present disclosure, the non-thermal treatment system comprises a second switch 420 upstream of switch 410 and configured to disrupt or create an electrical connection to the generator and an electrical connection to the applicator to turn the non- thermal treatment system off or on, respectively, as shown in FIG.1B. In an aspect, switch 420 is a mechanical switch. In an aspect, switch 420 is an electronic switch. In an aspect, switch 420 is a latch switch. In an aspect, switch 420 is a momentary switch. In a further aspect, switch 420 is a double pole double throw switch, a single pole double throw switch, or a changeover switch. In an aspect of the present disclosure, the non-thermal treatment system comprises switch 410, switch 420, or both switch 410 and switch 420. In an aspect of the present disclosure, switch 410 is configured to disrupt the electrical connection to the generator and an electrical connection to the applicator to turn the non-thermal treatment system off.

[0137] In an aspect, the present disclosure provides for and includes an applicator comprising a first electrode 210 and a second electrode 220, as depicted in FIGS.1A and 1B, which can be used in a bipolar or monopolar mode for the application of an electric field. In one aspect, the applicator is a clamping probe 201. Referring to FIG.2B, clamping probe 201 can be applied as a clamp with one prong of the forceps as a first electrode 210 and the other prong of the forceps as a second electrode 220. In an aspect, the clamping probe are utilized as a blunt tip to apply the energy from the generator to a surgical site in a monopolar mode. In an aspect, the clamping probe 201 is used in monopolar mode with a dispersive pad applied to a patient in need thereof to complete the circuit and to ablate endometriosis lesions. In an aspect, the clamping probe is used in bipolar mode, with each electrode connected to the two outputs of the generator, to grasp the tissue and apply the energy between the first and second electrodes to treat a patient in need thereof. In an aspect, the two prongs of clamping probe 201 have an inner jagged edge and smooth outer edge. Without limiting the disclosure, an exemplary embodiment of clamping probe 201 is shown in FIGS.87A and 88A (“BiClamp5”).

[0138] In an aspect of the specification, applicator 200 is a clamping probe 301. In an aspect, the clamping probe 301 has bilateral curved clamps narrowing in width. Referring to FIG.88B, clamping probe 301 can be applied as a clamp with one prong of the clamp as a first electrode 310 and the other prong of the clamp as a second electrode 320. In an aspect, the clamping probe are utilized as a blunt tip to apply the energy from the generator to a surgical site in a monopolar mode.Arnold & Porter Ref. P35370WO00 In an aspect, the clamping probe 301 is used in monopolar mode with a dispersive pad applied to a patient in need thereof to complete the circuit and to ablate endometriosis lesions. In an aspect, the clamping probe is used in bipolar mode, with each electrode connected to the two outputs of the generator, to grasp the tissue and apply the energy between the first and second electrodes to treat a patient in need thereof. In an aspect, the two prongs of clamping probe 301 have an inner jagged edge and smooth outer edge. Without limiting the disclosure, an exemplary embodiment of clamping probe 301 is shown in FIGS.87B and 88B (“Maryland1”).

[0139] In an aspect of the present disclosure, applicator 200 is a clamping probe 401. In an aspect, clamping probe 401 comprises a first clamp with a jagged edge and a second clamp with a smooth edge. In an aspect, clamping probe 401 comprises a first clamp with a jagged edge and a second clamp with a smooth edge that is curved at the tip. In an aspect, the first clamp and second clamp of clamping probe 401 are open in the center of both clamps, and the second clamp has a jagged cross section (e.g., FIG.80). Referring to FIG.87C, clamping probe 401 can be applied as a clamp with one prong of the clamp as a first electrode 410 and the other prong of the clamp as a second electrode 420. In an aspect, the clamping probe is utilized as a blunt tip to apply the energy from the generator to a surgical site in a monopolar mode. In an aspect, the clamping probe 401 is used in monopolar mode with a dispersive pad applied to a patient in need thereof to complete the circuit and to ablate endometriosis lesions. In an aspect, the clamping probe is used in bipolar mode, with each electrode connected to the two outputs of the generator, to grasp the tissue and apply the energy between the first and second electrodes to treat a patient in need thereof. Without limiting the disclosure, an exemplary embodiment of clamping probe 401 is shown in FIG.87C (“BiClamp2”).

[0140] In an aspect, the present disclosure provides for and includes the articulating tip applicator 202. Referring to FIGS.2A, 2B, 3A, and 3B, an articulating tip applicator 202 (also referred to as an articulating tip applicator) comprises a shaft comprising: a distal end, a flexible section, and a proximal end. In an aspect, the distal end comprises a first electrode 210 comprising a first stainless steel rod 212, a distal end 213, and a proximal end 214; a second electrode 220 comprises a second stainless steel tube 222, a distal end 223, and a proximal end 224; an insulating layer 250; and a flexible section 240. In an aspect, the flexible section is situated between the proximal and distal ends of the shaft. In an aspect, the proximal end of the shaft comprises mechanical levers connected to the flexible section with cables. In an aspect, the first electrode 210 and the second electrode 220 are concentric, and the proximal end 214 of the first electrode and the distal end 223 of the second electrode are separated by a distance in length 230w. In an aspect of the present disclosure, the insulating layer is not exposed when the device is in monopolar mode.Arnold & Porter Ref. P35370WO00 In an aspect, the exposed insulating layer equals the distance in length between the proximal end of the first electrode and the distal end of the second electrode 230w.

[0141] As depicted in FIG.3A, in one aspect, first electrode 210 and second electrode 220 have a cylindrical shape. In an aspect, first electrode 210 and second electrode 220 have a frustoconical shape. In a further aspect, first electrode 210 and second electrode 220 have a cylindrical shape, a frustoconical shape or any other shape without departing from the scope of the present disclosure. In another aspect of the present disclosure, a stainless steel tube 222 is a stainless steel hollow rod 222.

[0142] In an aspect of the present disclosure, the distance in length 230w between the proximal end 214 of the first electrode and the distal end 223 of the second electrode is adjustable. In an aspect, the distance in length 230w manually adjustable. In an aspect, the distance in length 230w is electronically adjustable. In an aspect, the distance in length is adjustable based on a feedback sensor as described in the present disclosure. For example, FIG.3A depicts an embodiment where 230w is shorter compared to 230w in FIG.3B. In an aspect, the distance in length 230w is between 0.75 mm and 2.25 mm. In an aspect, the distance in length 230w is at least 0.25 mm, at least 0.5 mm, at least 0.75 mm, at least 1 mm, at least 1.25 mm, at least 1.5 mm, at least 1.75 mm, at least 2 mm, at least 2.25 mm, or at least 2.5 mm. In an aspect, the distance in length 230w is between 0.25 and 2.5 mm or between 0.5 and 2 mm.

[0143] In one aspect, a first outer diameter 216 of the first electrode 210 is between 3 mm and 9 mm. In an aspect, the first outer diameter 216 of the first electrode 210 is at least 3 mm, at least 4 mm, at least 5 mm, at least 6 mm, at least 7 mm, or at least 8 mm. In an aspect, the first outer diameter 216 is between 4 and 8 mm or between 5 and 7 mm. In an aspect, the first outer diameter 216 of the first electrode 210 is 6 mm.

[0144] In other aspects, a second outer diameter 226 of the second electrode 220 is between 1 mm and 5 mm. In an aspect, the second outer diameter 226 of the second electrode 220 is at least 1 mm, at least 2 mm, at least 3 mm, or at least 4 mm. In a further aspect, the first outer diameter 226 is between 1 and 4 mm or between 2 and 4 mm. In an aspect, the second outer diameter 226 of the second electrode 220 is 3 mm.

[0145] In aspects of the present disclosure, articulating tip applicator 202 comprises a first stainless steel rod 212 surrounded by a first insulating layer 212i between the first electrode 210 and the second electrode 220, and a second stainless steel tube 222 surrounded by a second insulating layer 222i. As depicted in FIGS.3A and 3B, in an embodiment, first electrode 210 comprises the first stainless steel rod 212 that extends towards a distal end of the articulating tip applicator 202. For example, in aspects of the present disclosure, as depicted in FIGS.3A and 3B,Arnold & Porter Ref. P35370WO00 first electrode 210 is configured to telescopingly extend away from second electrode 220 along an axis (“A” in FIG.2A) towards a distal end of the articulating tip applicator 202. In an aspect, first electrode 210 may be slidably movable with respect to second electrode 220 between a retracted position (shown in FIG.3A) and an extended position (shown in FIG.3B). For example, a distance in length 230w between the proximal end of the first electrode 214 and the distal end 223 of the second electrode may be selected from the group consisting of between 0.75 mm to 2.25 mm, at least 0.75 mm, and at most 2.25 mm.

[0146] FIGS.4A and 4B depict simulated surface electric potential graphs (in Volts) of non- thermal treatment system in two different modes, i.e., bipolar mode 510 and monopolar mode 520. As depicted in FIG.4A, in an aspect, switch 400 is configured to change electrical connection with the generator 114 when supplying a different catheter voltage. In an aspect, the switch is capable of changing positions between a first position (shown in FIG.4A) to apply an electric field of a first mode 510 through the applicator and a second position (shown in FIG.1) to apply an electric field of a second mode 520 through the applicator. For example, switch 400 is capable of changing catheter voltage between bipolar mode 510 and monopolar mode 520 through a generator GUI 530. In an aspect, bipolar mode 510 may be advantageous for treatment with precise application of IRE dose. Since strength of electric field decreases rapidly as tissue positions away from two electrodes, bipolar mode comprises a safer mode when the treatment is near critical structures. Additionally, a maximum voltage is limited to prevent dielectric breakdown of the insufflating gas (for laparoscopy) or air (open).

[0147] In a bipolar mode, as depicted in FIG.4A, a surface electric potential between the first electrode 210 and the second electrode 220 comprises a range between 500 and 800 V. In an aspect, the surface electric potential between the first electrode 210 and the second electrode 220 is at least 500 V, at least 600 V, at least 700, or at least 800. In a further aspect, the surface electric potential of the second electrode 220 is between 600 and 700 V.

[0148] In a monopolar mode, as depicted in FIG.4B, a surface electric potential between the first electrode 210 (or the second electrode 220) and the dispersive pad 300 comprises a range between 500 and 2000 V. In an aspect, the surface electric potential between the first electrode 210 (or the second electrode 220) and the dispersive pad 300 is at least 500 V, at least 1000 V, at least 1500, or at least 2000. In a further aspect, the surface electric potential of the second electrode 220 is between 500 and 1500 V.

[0149] Compared to the bipolar mode, monopolar mode may produce lesions that are deeper since the voltage is not limited by dielectric breakdown. Particularly, the dispersive pad 300 (electrode) may be placed on patient’s skin and includes much larger surface area than combinationArnold & Porter Ref. P35370WO00 of first electrode’s 210 surface area and second electrode’s 220 surface area. A dispersive pad is an electrode with a relatively large surface area which is positioned on the patient as it completes the circuit from an active electrode (e.g., first or second electrode 210, 220). The dispersive pad allows the high frequency current to flow back with a low current intensity thereby preventing any physical effects, such as undesired burns.

[0150] In some aspects, monopolar pad is replaced by a large surface electrode such as a conductive surface on a uterine manipulator during a laparoscopic procedure.

[0151] FIGS.5-7 depict an exemplary electrical voltage / current profile for an activating, resting and / or modulating impulse applied to lesions (e.g. endometrial lesions) in accordance with an embodiment of the present disclosure. As shown in FIG.5, current 501 and voltage 502 refer to those that are produced within the patient by the electrodes. As shown in FIGS.5-7, a suitable electrical voltage / current profile 500 for the modulating impulses to lesions is achieved using pulse generator 110 (See FIG.1). The pulse generator uses a power source 110 and a control unit comprising, for instance, a processor, a clock, a memory, or the like, to produce a pulse pattern 505 to the electrodes 210, 220 (in FIG.1) that deliver the modulating impulse 504 to the lesions. The parameters of the modulation signal, such as the frequency, amplitude, duty cycle, pulse width, pulse shape, etc., are programmable. An external communication device may modify the pulse generator programming to facilitate treatment.

[0152] FIG.6 depicts an oscilloscope recording of voltage and current for a short application. For example, as shown in FIG.6, when activated by a switch such as a foot switch, a burst 525 includes 4 packets (each packet 522), and groups of packets are applied twice. In an aspect, the burst 525 is followed by a period with no signal, dead time D (the inter-burst period). The dead time D is between groups of bursts. The dead time is between 0.1 and 1 seconds. In a further aspect, a foot switch turns ON and OFF the pattern of a burst. In an aspect, pattern of a burst followed by dead time D (i.e., inter-burst period) repeats itself with a period of T.

[0153] As depicted in FIG.6, bursts of wave – between a quasi-square wave to sinusoidal – pulses are determined to be a particular treatment waveform. Each packet 522 has one or more quasi-square wave to sinusoidal cycles (pulses) 504 with a period of tau (τ). For example, each packet 522 has a 10 cycles per packet, i.e., M=10. Each burst 525 has one or more sinusoidal packets 522 with a rest period I. For example, each burst 525 has 4 sinusoidal packets 522, i.e., N=4. The rest period(r) is between 0.001 and 0.01 seconds.

[0154] FIG.7 depicts another oscilloscope recording of voltage and current for a long application. For example, as shown in FIG.7, when activated by a switch such as a foot switch,Arnold & Porter Ref. P35370WO00 and various groups of packets are applied with a burst 525 that includes 4 packets with no signal, dead time D (the inter-burst period).

[0155] In an aspect of the present disclosure, a generator is capable of applying an electric field to an endometrial lesion through one or more electrodes. In an aspect, a voltage from the generator is between 500 and 3,000 volts for treatment of a lesion. In an aspect, the voltage from the generator is at least 500 V, at least 1000 V, at least 1500 V, at least 2000 V, at least 2500 V, or at least 3000 V. In a further aspect, the voltage from the generator is between 1000 and 2500 V or between 1500 and 2000 V. In an aspect, a pulse from the generator is between 25 and 200 µs. In an aspect, the pulse from the generator is at least 25 µs, at least 50 µs, at least 100 µs, at least 150 µs, or at least 200 µs. In a further aspect, the pulse from the generator is between 50 and 175 µs or between 75 and 150 µs. FIG.7B is an example of another oscilloscope recording for a long application.

[0156] Referring to FIGS.45-50B, a non-thermal treatment system 100 of the present disclosure is shown. In various embodiments, the non-thermal treatment system 100 may comprise an alternative applicator 1200 (i.e., instead of applicator 200) for non-thermal treatment of one or more lesions of a patient.

[0157] In an aspect, the present disclosure provides for and includes an applicator comprising a first electrode 1210 and a second electrode 1220, as depicted in FIGS.45-50B. In an aspect, the first electrode and second electrode can be used in bipolar or monopolar mode for the application of an electric field.

[0158] As depicted in FIG.45A, in an aspect, the present disclosure provides for and includes a catheter 1202. The catheter 1202 (also referred to as an articulating tip applicator) comprises a proximal end 1201, a shaft 1203, second applicator (tip) 1205 comprising a vaulted (or dome-like) shape, and a distal end 1209. In an aspect, the present disclosure provides for an applicator (tip) 1205 comprising an opening 1207 at distal end 1209. FIGS.45B and 45C depict two-dimensional schematics of catheter 1200. The design is configured to make the tissue contact area less sensitive to change in signature and treatment zones. As depicted in FIGS.45B and 45C, the second applicator (tip) 1205 comprises a first electrode 1210 and a second electrode 1220 separated by an insulating layer 1290 in distal end 1209. In an aspect, as depicted in FIG.45B, the first electrode 1210 and the second electrode 1220 extend axially and are separated by a distance in length 1230w therebetween, for example, between the second end 1214 of the first electrode and the first end 1223 of the second electrode. In an aspect the distance in length 1230w is between 1.5 mm and 2.5 mm. In an aspect, the distance in length 1230w is at least 1.5 mm, at least 2 mm, or at least 25 mm. In a further aspect, the distance in length 1230w is between 1.5 mm and 2 mm or between 2 mmArnold & Porter Ref. P35370WO00 and 2.5 mm. In an aspect, distal end 1209 comprises various segments (e.g., a set of first electrode 1210 and second electrode 1220 separated by an insulating layer) between 4 and 12 segments. In an aspect, there are at least 4, at least 8, or at least 12 segments. In a further aspect, there are between 4 and 6, between 4 and 8, between 5 and 7, between 6 and 12, 8 and 12, or between 10 and 12.

[0159] In an aspect and as depicted in FIG.45C, adjacent to the opening 1207, the first electrode 1210 and the second electrode 1220 extend axially and are separated by an angle 1230θ therebetween, for example, between the second end 1214 of the first electrode and the first end 1223 of the second electrode. In an aspect, first electrode 1210 and second electrode 1220 have a substantially frustoconical surface shape. In a further aspect, first electrode 1210 and second electrode 1220, comprise a substantially frustoconical surface shape or any other shape without departing from the scope of the present disclosure.

[0160] FIGS.46-47 depict computer simulations of IRE zone created as the tip changes angle. For example, in an aspect, FIGS.46 (A) and (C) depict an embodiment where IRE zone created 1297a, 1297c has a vaulted (or dome-like) shape compared to a substantially even depth of IRE zone created 1297b in FIG.46 (B) due to an associated angle that the tip applied to tissue layers. FIGS.46 (A) and (C) depict an embodiment where IRE zone created 1297a, 1297c has a deeper area (or larger depth) compared to IRE zone created 1297b in FIG.46 (B) due to an opening 1207 shown in FIGS.46 (D) and (E) and an associated angle that the tip applied to tissue layers.

[0161] In an aspect, as depicted in FIG.47 (D), IRE zone created 1297d shows a different depth in tissue due to an opening 1207 shown in FIGS.46I) and (E) and an associated angle that the tip is applied to tissue layers.

[0162] Referring to FIGS.48A, 48B, and 48C, the second applicator (tip) 1205 comprises an opening 1207 with an opening diameter 1226. In various embodiments, the opening diameter 1226 of the opening 1207 may be adjusted to make a tissue contact area less sensitive to change in signature and treatment zone. In an aspect of the present disclosure, an opening diameter 1226 of the opening 1207 is adjustable. In an aspect, the opening diameter 1226 is manually adjustable. In an aspect, the opening diameter 1226 is electronically adjustable. In an aspect, the opening diameter 1226 is adjustable based on a feedback sensor as described in the present disclosure. For example, FIG.48B depicts an embodiment where opening 1207b has a larger opening diameter 1226b compared to that of opening 1207a in FIG.48A. As a further example, FIG.48C depicts an embodiment where opening 1207c has a smaller opening diameter 1226c compared to that of opening 1207a in FIG.48A. In an aspect, the opening diameter 1226 is between 1 mm and 3 mm. In an aspect, the opening diameter 1226 is at least 1 mm, at least 1.25 mm, at least 1.5 mm, at leastArnold & Porter Ref. P35370WO00 1.75 mm, at least 2 mm, at least 2.25 mm, at least 2.5 mm, at least 2.75 mm, or at least 3 mm. In an aspect, the opening diameter 1226 is between 1 and 2 mm. In an aspect, the opening diameter 1226 is between 1.25 and 2.75 mm. In an aspect, the opening diameter 1226 is between 1.25 and 2.5 mm. In an aspect, the opening diameter 1226 is between 1.25 and 2 mm. In an aspect, the opening diameter 1226 is between 1.5 mm and 2.5 mm. In an aspect, the opening diameter 1226 is between 1.5 and 2.75 mm. In a preferred embodiment, an opening diameter 1226 of the opening 1207 is in a range between 1 mm and 1.5 mm.

[0163] Referring to FIGS.45A, 45B, and 45C, in one aspect, a first outer diameter 1216 is between 1 mm and 3 mm. In an aspect, the first outer diameter 1216 of the first electrode 1210 is at least 1 mm, at least 1.25 mm, at least 1.5 mm, at least 1.75 mm, at least 2 mm, at least 2.25 mm, at least 2.5 mm, at least 2.75 mm, or at least 3 mm. In an aspect, the first outer diameter 1216 is between 1 and 3 mm. In an aspect, the first outer diameter 1216 is between 1.25 and 2.75 mm. In an aspect, the first outer diameter 1216 is between 1.25 and 2.5 mm. In an aspect, the first outer diameter 1216 is between 1.25 and 2 mm. In an aspect, the first outer diameter 1216 is between 1.5 mm and 2.5 mm. In an aspect, the first outer diameter 1216 is between 1.5 and 2.75 mm. In yet another aspect, the opening diameter 1226 is between 2 and 3 mm. In an aspect, opening diameter 1226 is smaller than a first outer diameter 1216.

[0164] FIGS.49A, 49B, and 49C provide additional embodiments of the present disclosure. Referring to FIGS.49A, 49B, and 49C, in an aspect, the second applicator (tip) 1205 comprises an insulating layer 1290 with an angle 1230θ between the first electrode 1210 and the second electrode 1220. In various embodiments, the angle 1230θ of the insulating layer 1290 may be selected to make tissue contact area less sensitive to change in signature and treatment zone. In some aspects, the angle 1230θ is adjustable based on a feedback information as described in the present disclosure. For example, FIG.49B depicts an embodiment where insulating layer 1290 has a bigger angle 1230θb compared to that of insulating layer 1290 in FIG.49A. In an aspect, FIG. 49C depicts an embodiment where insulating layer 1290 has a smaller angle 1230θc compared to that of insulating layer 1290 in FIG.48A. In an aspect, when the second applicator (tip) 1205 comprises two first electrodes 1210 and two second electrodes 1220, the angle 1230θ of the insulating layer 1290 is between 0 degree and 90 degree. In an aspect, the angle 1230θ is at least 0 degree, at least 15 degree, at least 30 degree, at least 45 degree, at least 60 degree, at least 75 degree, at least 90 degree. In an aspect, the opening diameter 1226 is between 15 and 75 degree or between 30 and 60 degree.

[0165] Referring to FIGS.49D-49E, the second applicator (tip) 1205 comprises one or more first electrodes 1210 and second electrodes 1220. For example, as depicted in FIG.49D, in anArnold & Porter Ref. P35370WO00 aspect, the second applicator (tip) 1205 comprises 4 first electrodes 1210a, 1210b, 1210c, 1210d and 4 second electrodes 1220a, 1220b, 1220c, 1220d. For another example, as depicted in FIG. 49E, in an aspect, the second applicator (tip) 1205 comprises a single first electrode 1210e and a single second electrode 1220e separated by an insulating layer 1290.to here

[0166] FIGS.50A and 50B depict simulated surface electric potential graphs (in Volts) of non- thermal treatment system an alternative applicator 1200 (i.e., as opposed to applicator 200) in two different modes, i.e., bipolar mode 1510 and monopolar mode 1520. As depicted in FIG.50A, in an aspect, switch 400 is configured to change electrical connection with generator 114 when supplying a different catheter voltage. In an aspect, the switch is capable of changing positions between a first position (shown in FIG.50A) to apply an electric field of a first mode 1510 through the applicator and a second position (shown in FIG.1) to apply an electric field of a second mode 1520 through the applicator. For example, switch 400 is capable of changing catheter voltage between bipolar mode 1510 and monopolar mode 1520 through a generator GUI 530. In an aspect, bipolar mode 1510 may be advantageous for treatment with precise application of IRE dose. Since strength of electric field decreases rapidly as tissue positions away from two electrodes, bipolar mode comprises a safer mode when the treatment is near critical structures. Additionally, a maximum voltage is limited to prevent dielectric breakdown of the insufflating gas (for laparoscopy) or air (open).

[0167] In a bipolar mode, as depicted in FIG.50A, a surface electric potential between the first electrode 1210 and the second electrode 1220 comprises a range between 500 and 800 V. In an aspect, the surface electric potential between the first electrode 1210 and the second electrode 1220 is at least 500 V, at least 600 V, at least 700, or at least 800. In a further aspect, the surface electric potential of the second electrode 1220 is between 600 and 700 V.

[0168] In a monopolar mode, as depicted in FIG.50B, a surface electric potential between the first electrode 1210 (or the second electrode 1220) and the dispersive pad 300 comprises a range between 500 and 2000 V. In an aspect, the surface electric potential between the first electrode 1210 (or the second electrode 1220) and the dispersive pad 300 is at least 500 V, at least 1000 V, at least 1500, or at least 2000. In a further aspect, the surface electric potential of the second electrode 220 is between 500 and 1500 V.

[0169] The present disclosure provides for and includes methods of non-thermal treatment of an endometrial lesion in a subject in need thereof. In an aspect, treatment methods comprise identifying the location, size and volume of the endometrial lesion in the subject. In an aspect, the methods also comprise determining an electric field useful in the treatment of the identified endometrial lesion and applying the electric field to the endometrial lesion for an appropriate timeArnold & Porter Ref. P35370WO00 and appropriate voltage using the non-thermal treatment system provided herewith. In an aspect, the present disclosure provides for and includes methods for causing controlled cell necrosis in an endometrium of a subject in need thereof. In an aspect, the treatment methods comprise determining an electric field useful in the treatment of an endometrial lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the endometrial lesion with a non-thermal treatment system and applicators as provided in the present disclosure. In an aspect of the present disclosure, the methods provided include an electric field that does not cause damage outside of the treatment area. In an aspect, the electric field is applied in cycles separated by one or more rest periods to reduce damage to healthy tissue. In an aspect, the number of cycles are selected from the group consisting of at least 5 cycles, at least 10 cycles, at least 15 cycles, at least 20 cycles, at least 25 cycles, at least 30 cycles, at least 35 cycles, at least 40 cycles, at least 45 cycles, and at least 50 cycles. In an aspect, the number of cycles are between 5 and 15, between 5 and 25, between 5 and 50, or between 5 and 100 cycles. In an aspect, the number of cycles is less than 150, less than 100, less than 50, or less than 25 cycles.

[0170] In an aspect of the present disclosure, the electric field is determined by the application of a pre-pulse followed by a fixed dose.

[0171] The present disclosure provides for, and includes, a non-thermal treatment system and devices for the treatment of a subject in need thereof. In an aspect of the present disclosure, a subject in need thereof is treated with a non-thermal treatment system and devices in monopolar mode. In an aspect of the present disclosure, a subject in need thereof is treated with a non-thermal treatment system and devices in a bipolar mode. In an aspect of the present disclosure, a subject in need thereof is an endometriosis patient. In an aspect, a subject in need thereof is a patient with lesions. In an aspect, a subject in need thereof is a cancer patient. In an aspect, a subject in need thereof is a patient with liver cancer, pancreatic cancer, kidney cancer, lung cancer, or a combination thereof. In an aspect, a subject in need thereof is a patient with one or more tumors. In an aspect, a subject in need thereof is a patient with a endometrial lesion, a malignant skin lesion, a non-malignant skin lesion, a bladder lesion, a ductal breast cancer lesion, a lesion of the gastrointestinal system, or a combination thereof. An aspect of the present disclosure provides for, and includes, a non-thermal treatment system and devices for the treatment of a lesion selected from the group consisting of endometrial lesion, a malignant skin lesion, a non-malignant skin lesion, a bladder lesion, a ductal breast cancer lesion, a lesion of the gastrointestinal system, lesions of the esophagus, or a combination thereof. In an aspect, the present disclosure provides for, and includes, a non-thermal treatment system and devices for the treatment of Barrett’s esophagus, polyps, malignant lesions that are not transmural, or a combination thereof.Arnold & Porter Ref. P35370WO00

[0172] As used herein the term “about” refers to ± 10 %.

[0173] The terms “comprises,” “comprising,” “includes,” “including,” “having,” and their conjugates mean “including but not limited to.”

[0174] The term “consisting of” means “including and limited to.”

[0175] The term “consisting essentially of” means that the composition, method or structure may include additional ingredients, steps and / or parts, but only if the additional ingredients, steps and / or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.

[0176] As used herein, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a compound” or “at least one compound” may include a plurality of compounds, including mixtures thereof.

[0177] Throughout this application, aspects of this disclosure may be presented in a range format. The description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as “from 1 to 6” should be considered to have specifically disclosed subranges such as “from 1 to 3,” “from 1 to 4,” “from 1 to 5,” “from 2 to 4,” “from 2 to 6,” “from 3 to 6,” etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0178] Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases “ranging / ranges between” a first indicate number and a second indicate number and “ranging / ranges from” a first indicate number “to” a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.

[0179] As used herein the term “method” refers to manners, means, techniques, and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques, and procedures either known to or readily developed from known manners, means, techniques, and procedures by practitioners of the chemical, pharmacological, biological, biochemical, and medical arts.

[0180] As used herein the term “exemplary” means that it’s an example or illustration, not necessarily the best mode.

[0181] The present disclosure provides for the following embodiments:

[0182] Embodiment 1. A device for non-thermal treatment of one or more lesions in a subject in need thereof comprising: a generator 110 comprising a first output connector and a second output connector; an applicator 200, wherein applicator 200 is a articulating tip applicatorArnold & Porter Ref. P35370WO00 202 comprising a first electrode 210 and a second electrode 220; a dispersive pad 300; and a switch 410 in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator.

[0183] Embodiment 2. The device of embodiment 1, wherein the switch 410 is a mechanical switch or an electronic switch.

[0184] Embodiment 3. The device of embodiment 1, wherein the switch 410 is a latch or a momentary switch.

[0185] Embodiment 4. The device of embodiment 1, wherein the switch 410 is a double pole double throw switch, a single pole double throw switch or a changeover switch.

[0186] Embodiment 5. The device of any one of embodiments 1 to 4, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad.

[0187] Embodiment 6. The device of any one of embodiments 1 to 5, wherein the switch 410 in the first position is configured to provide an electrical connection between the generator 110 through the first output connector with the first electrode 210 and the second electrode 220 of the applicator and provide an electrical connection between the generator through the second output connector with the dispersive pad 300.

[0188] Embodiment 7. The device of any one of embodiments 1 to 6, wherein the first electrode 210 and the second electrode 220 are in electrical connection with each other.

[0189] Embodiment 8. The device of any one of embodiments 1 to 7, wherein the dispersive pad 300 is configured to be in contact with the patient when the electric field is applied.

[0190] Embodiment 9. The device of any one of embodiments 1 to 8, wherein the applicator 200 is configured to operate as a blunt tip for the treatment of the one or more lesions.

[0191] Embodiment 10. The device of any one of embodiments 1 to 9, wherein the dispersive pad 300 is a large surface electrode.

[0192] Embodiment 11. The device of any one of embodiments 1 to 10, wherein the large surface electrode comprises a surface area between 100 cm2 and 200 cm2.

[0193] Embodiment 12. The device of any one of embodiments 1 to 11, wherein the second mode is a bipolar mode that generates an electric field between the first electrode 210 and the second electrode 220 of the applicator.

[0194] Embodiment 13. The device of any one of embodiments 1 to 12, wherein the switch in the second position is configured to provide an electronic connection between the generator 110 through the first output connector with the first electrode 210 and provide an electronic connection between the generator 110 through the second output connector with the second electrode 220.Arnold & Porter Ref. P35370WO00

[0195] Embodiment 14. The device of any one of embodiments 1 to 13, wherein there is no electrical connection between the generator 110 and the dispersive pad 300.

[0196] Embodiment 15. The device of any one of embodiments 1 to 14, wherein the one or more lesions of the patient comprise one or more endometrial lesions.

[0197] Embodiment 16. The device of any one of embodiments 1 to 15, wherein the electric field of the first mode through the applicator 300 is capable of forming a treatment zone comprising the one or more lesions.

[0198] Embodiment 17. The device of any one of embodiments 1 to 16, wherein the electric field of the second mode through the applicator is capable of forming a treatment zone comprising the one or more lesions.

[0199] Embodiment 18. The device of any one of embodiments 1 to 17, wherein the first electrode 210 comprises a first stainless steel rod 212 surrounded by a first insulating layer 212i, a distal end 213, and a proximal end 214, wherein the second electrode 220 comprises a second stainless steel tube 222 surrounded by a second insulating layer 222i, a distal end 223, and a proximal end 224, wherein the first electrode 210 and the second electrode 220 are concentric, and wherein the proximal end 214 of the first electrode and the distal end 223 of the second electrode are separated by a distance in length 230w.

[0200] Embodiment 19. The device of any one of embodiments 1 to 18, further comprising an insulating layer 230 between the first electrode 210 and the second electrode 220.

[0201] Embodiment 20. The device of any one of embodiments 1 to 19, wherein the insulating layer 212 extends beyond the distal end of the first electrode by between 0 and 2 mm.

[0202] Embodiment 21. The device of any one of embodiments 1 to 20, wherein the first electrode comprises a first outer diameter 216 of between 2 and 4 millimeters (mm).

[0203] Embodiment 22. The device of any one of embodiments 1 to 21, wherein the first outer diameter 216 is approximately 3 mm.

[0204] Embodiment 23. Device of any one of embodiments 1 to 22, wherein the second electrode 220 comprises a second outer diameter 226 of between 6 and 10 mm.

[0205] Embodiment 24. The device of any one of embodiments 1 to 22, wherein the second outer diameter 226 is approximately 6 mm.

[0206] Embodiment 25. The device of any one of embodiments 1 to 24, wherein the first electrode 210 comprises a wall thickness between 0 mm and 0.8 mm.

[0207] Embodiment 26. The device of any one of embodiments 1 to 25, wherein the second electrode 220 comprises a wall thickness of at least 1 mm.Arnold & Porter Ref. P35370WO00

[0208] Embodiment 27. The device of any one of embodiments 1 to 26, wherein the distance in length between the proximal end 214 of the first electrode and the distal end 223 of the second electrode is between 0 mm and 6 mm.

[0209] Embodiment 28. The device of any one of embodiments 1 to 27, wherein the distance in length 230w between the proximal end 214 of the first electrode and the distal end 223 of the second electrode is adjustable.

[0210] Embodiment 29. The device of any one of embodiments 1 to 28, wherein the distance in length 230w between proximal end 214 of the first electrode and the distal end 223 of the second electrode is between 0.75 mm to 2.25 mm.

[0211] Embodiment 30. The device of any one of embodiments 1 to 29, wherein the distance in length 230w between the proximal end 214 of the first electrode and the distal end 223 of the second electrode is at least 0.75 mm.

[0212] Embodiment 31. The device of any one of embodiments 1 to 30, wherein the distance in length 230w between the proximal end 214 of the first electrode and the distal end 223 of the second electrode is at most 6 mm.

[0213] Embodiment 32. The device of any one of embodiments 1 to 31, wherein the distance in length 230w between the proximal end 214 of the first electrode and the distal end 223 of the second electrode is between 0 and 6 mm.

[0214] Embodiment 33. A device for an application of a non-thermal treatment dose comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a first electrode and a second electrode; a first switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply a therapeutic electric field of a first mode through the applicator and a second position to apply a therapeutic electric field of a second mode through the applicator; a dispersive pad in an electrical connection with the second output connector; a sensor configured to receive a first signal before the application of a non-thermal treatment dose and a second signal after the application of the non- thermal treatment dose; a processor electronically coupled to the switch and the sensor, a non- transitory computer readable medium electronically coupled to the processor, wherein the non- transitory computer readable medium comprises instructions stored thereon that, when executed on the processor, perform the steps of receiving the first signal from the sensor and the second signal from the sensor; and determining a difference between the first signal and the second signal, comparing the difference against a predetermined threshold; and instructing the application to end if the difference exceeds the predetermined threshold.Arnold & Porter Ref. P35370WO00

[0215] Embodiment 34. The device of embodiment 33, wherein the first switch is in a first electrical connection with the first output connector and in a second electrical connection with the first electrode 210.

[0216] Embodiment 35. The device of any one of embodiments 33 to 34, wherein the instructing the application to end comprises a third signal to a second switch 420 upstream of the first switch and in a first electrical connection with the first output connector and in a second electrical connection with the first electrode, wherein the second switch is configured to disrupt the first electrical connection to the first output connector and the second electrical connection with the first electrode.

[0217] Embodiment 36. The device of any one of embodiments 33 to 35, wherein the second switch is an automatic switch or a manual switch.

[0218] Embodiment 37. The device of any one of embodiments 33 to 36, wherein the first switch is an automatic switch or a manual switch.

[0219] Embodiment 38. A device for an application of a non-thermal treatment dose comprising: a generator 110 comprising a first output connector and a second output connector; an applicator 200 comprising a first electrode 210 in a first electrical connection with the first output and a second electrode 220 in a second electrical connection with the second output; a switch in a first electrical connection with the first output connector and in a second electrical connection with the first electrode, wherein the switch in a first position is configured to apply a therapeutic electric field through the first electrode, and wherein the switch in a second position is configured to disrupt the first electrical connection to the first output connector and the second electrical connection with the first electrode; a sensor configured to receive a first signal before the application of a non-thermal treatment dose and a second signal after the application of the non- thermal treatment dose; a processor electronically coupled to the switch and the sensor; a non-transitory computer readable medium electronically coupled to the processor, wherein the non-transitory computer readable medium comprises instructions stored thereon that, when executed on the processor, perform the steps of receiving the first signal from the sensor and the second signal from the sensor; and determining a difference between the first signal and the second signal, comparing the difference against a predetermined threshold; and instructing the switch to configure to the second position.

[0220] Embodiment 39. The device of embodiment 38, wherein the switch is an automatic switch or a manual switch.

[0221] Embodiment 40. A device for non-thermal treatment comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a firstArnold & Porter Ref. P35370WO00 electrode and a second electrode; a dispersive pad; and a first switch 410 in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator; a second switch 420 upstream of the first switch and in a first electrical connection with the first output connector and in a second electrical connection with the first electrode, wherein the second switch is configured to disrupt the first electrical connection to the first output connector and the second electrical connection with the first electrode; a sensor configured to recognize a signal; a processor electronically coupled to the switch and configured to receive a second signal from the sensor; and a non-transitory computer readable medium electronically coupled to the processor, wherein the non-transitory computer readable medium comprises instructions stored thereon activated, after the receipt of the signal, to perform the steps of disconnecting the electrical connection between the first electrical connection and the first output.

[0222] Embodiment 41. The device of embodiment 40, wherein the first switch is an automatic switch or a manual switch.

[0223] Embodiment 42. The device of embodiment 40, wherein the second switch is an automatic switch or a manual switch.

[0224] Embodiment 43. The device of any one of embodiments 33, 38, or 40, wherein the signal comprises a measurement for electrical magnitude and phase.

[0225] Embodiment 44. A method of non-thermal treatment of a lesion in a subject in need thereof comprising: identifying the location, size and volume of the lesion in the subject; determining an electric field useful in the treatment of the identified lesion; and applying the electric field to the lesion for an appropriate time and appropriate voltage using a device comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a first electrode and a second electrode; a dispersive pad; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator.

[0226] Embodiment 45. The method of embodiment 44, wherein the first mode is a monopolar mode that generates aa electric field between the applicator and the dispersive pad.

[0227] Embodiment 46. The method of any one of embodiments 44 to 45, wherein the dispersive pad is in contact with the subject in need thereof.

[0228] Embodiment 47. The method of any one of embodiments 44 to 46, wherein the lesion is endometrial lesion.Arnold & Porter Ref. P35370WO00

[0229] Embodiment 48. A method of non-thermal treatment of a lesion in a subject in need thereof comprising: determining an electric field useful in the treatment of the lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the lesion with a device comprising: a generator comprising a first output and a second output; an applicator comprising a first electrode and a second electrode; a dispersive pad; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply the electric field of the first mode through the applicator and a second position to apply the electric field of the second mode through the applicator, wherein the electric field is within a treatment area in the subject in need thereof.

[0230] Embodiment 49. The method of embodiment 48, wherein the lesion is an endometrial lesion, a malignant skin lesion, a non-malignant skin lesion, a bladder lesion, a ductal breast cancer lesion, a lesion of the gastrointestinal system, or a combination thereof.

[0231] Embodiment 50. The method of any one of embodiments 48 to 49, wherein the electric field does not cause damage outside of the treatment area.

[0232] Embodiment 51. The method of any one of embodiments 48 to 50, wherein the applying the determined electric field comprises applying the determined electric field in cycles separated by one or more rest periods to reduce thermal damage.

[0233] Embodiment 52. The method of any one of embodiments 48 to 51, wherein the cycles comprises between 0 and 150 cycles.

[0234] Embodiment 53. The method of any one of embodiments 48 to 52, wherein the cycles comprises at least 10 cycles.

[0235] Embodiment 54. The method of any one of embodiments 48 to 53, wherein the cycles comprises at least 15 cycles.

[0236] Embodiment 55. The method of any one of embodiments 48 to 54, wherein the cycles comprises at least 20 cycles.

[0237] Embodiment 56. The method of any one of embodiments 48 to 55, wherein the cycles comprises no more than 100 cycles.

[0238] Embodiment 57. The method of any one of embodiments 48 to 56, wherein the cycles comprises between 0 and 150 cycles.

[0239] Embodiment 58. The method of any one of embodiments 48 to 57, wherein the output frequency of the electric field comprises between 200 and 900 kilohertz (kHz).

[0240] Embodiment 59. The method of any one of embodiments 48 to 58, wherein the output frequency of the electric field is 250 kHz.Arnold & Porter Ref. P35370WO00

[0241] Embodiment 60. The method of any one of embodiments 48 to 59, wherein the cycles are 40 microseconds each.

[0242] Embodiment 61. The method of any one of embodiments 48 to 60, wherein the electric field comprises 500 to 2000 volts (V).

[0243] Embodiment 62. The method of any one of embodiments 48 to 61, wherein the electric field comprises 500 to 2000 volts (V).

[0244] Embodiment 63. The method of any one of embodiments 48 to 62, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad.

[0245] Embodiment 64. The method of any one of embodiments 48 to 63, wherein the voltage is from 500 to 2000 V.

[0246] Embodiment 65. The method of any one of embodiments 48 to 64, wherein the voltage is from 1500 to 2000 V.

[0247] Embodiment 66. The method of any one of embodiments 48 to 65, wherein the dispersive pad is configured to be in contact with the patient when the electric field is applied.

[0248] Embodiment 67. The method of any one of embodiments 48 to 66, wherein the second mode is a bipolar mode that generates an electric field between the first electrode and the second electrode of the applicator.

[0249] Embodiment 68. The method of any one of embodiments 48 to 67, wherein the voltage is from 500 to 800 V.

[0250] Embodiment 69. A method of causing controlled cell necrosis in an endometrium of a subject in need thereof comprising: determining an electric field useful in the treatment of an endometrial lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the endometrial lesion with a device comprising: a generator comprising a first output and a second output, an applicator comprising a first electrode and a second electrode; a dispersive pad; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply the electric field of the first mode through the applicator and a second position to apply the electric field of the second mode through the applicator, wherein the electric field of the first mode or the electric field of the second mode is within a treatment area of the subject in need thereof and causes cell necrosis within the treatment area.

[0251] Embodiment 70. The method of embodiment 69, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad.Arnold & Porter Ref. P35370WO00

[0252] Embodiment 71. The method of any one of embodiments 69 to 70, wherein the dispersive pad is configured to be in contact with the patient when the electric field is applied.

[0253] Embodiment 72. The method of any one of embodiments 69 to 71, wherein the second mode is a bipolar mode that generates an electric field between the first electrode and the second electrode of the applicator.

[0254] Embodiment 73. The method of any one of embodiments 44 to 72, wherein the determining an electric field comprises: applying the electric field in the second mode, wherein the second mode is a bipolar mode that generates an electric field between the first electrode and the second electrode of the applicator; and recording the magnitude and phase.

[0255] Embodiment 74. The method of any one of embodiments 48 to 73, wherein the determining an electric field comprises: applying the electric field in the first mode, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad; and recording the magnitude and phase.

[0256] Embodiment 75. The method of any one of embodiments 44 to 74, further comprising determining an application time for applying the electric field comprising: applying the electric field in the first mode, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad; and recording the magnitude and phase.

[0257] Embodiment 76. The method of any one of embodiments 44 to 75, further comprising determining an application time for applying the electric field comprising: applying the electric field in the second mode, wherein the second mode is a bipolar mode that generates an electric field between the first electrode and the second electrode of the applicator; and recording the magnitude and phase.

[0258] Embodiment 77. The method of any one of embodiments 44 to 76, wherein the determining an electrical field comprises a pre-pulse to determine contact followed by a fixed dose.

[0259] Embodiment 78. The device of any one of embodiments 1 to 43, wherein the applicator is an articulating tip catheter comprising a shaft comprising: a distal end comprising a first electrode 210 comprises a first stainless steel rod 212, a distal end 213, and a proximal end 214; a second electrode 220 comprises a second stainless steel tube 222, a distal end 223, and a proximal end 224, and an insulating layer; a flexible section; and a proximal end comprising mechanical levers connected to the flexible section with cables, wherein the flexible section is situated between the proximal end and the distal end, and wherein the first electrode 210 and the second electrode 220 are concentric, and the proximal end 214 of the first electrode and the distal end 223 of the second electrode are separated by a distance in length 230w.Arnold & Porter Ref. P35370WO00

[0260] Embodiment 79. An articulating tip applicator comprising: a shaft comprising: a distal end comprising a first electrode 210 comprises a first stainless steel rod 212, a distal end 213, and a proximal end 214; a second electrode 220 comprises a second stainless steel tube 222, a distal end 223, and a proximal end 224, and an insulating layer; a flexible section; and a proximal end comprising mechanical levers connected to the flexible section with cables, wherein the flexible section is situated between the proximal end and the distal end, and wherein the first electrode 210 and the second electrode 220 are concentric, and the proximal end 214 of the first electrode and the distal end 223 of the second electrode are separated by a distance in length 230w.

[0261] Embodiment 80. The articulating tip applicator of embodiment 79, wherein the proximal end of the shaft further comprises a motor.

[0262] Embodiment 81. The articulating tip applicator of any one of embodiments 79 to 80, wherein the proximal end of the shaft further comprises electrical switches.

[0263] Embodiment 82. The articulating tip applicator of any one of embodiments 79 to 81, wherein the shaft further comprises an electrical or mechanical rotation.

[0264] Embodiment 83. The articulating tip applicator of any one of embodiments 79 to 82, wherein a distal end is aligned along an axis of the shaft in a relaxed position.

[0265] Embodiment 84. A device for non-thermal treatment of one or more lesions in a subject in need thereof comprising: a generator 110 comprising a first output connector and a second output connector; an applicator 1200 comprising a first electrode 1210 and a second electrode 1220; a dispersive pad 300; and a switch 410 in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator.

[0266] Embodiment 85. The device of embodiment 84, wherein the switch 410 is selected from the group consisting of a mechanical switch, an electronic switch, a latch switch, a momentary switch, a double throw switch, a single pole switch, or a changeover switch.

[0267] Embodiment 86. The device of any one of embodiments 84 or 85, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad.

[0268] Embodiment 87. The device of any one of embodiments 84 to 86, wherein the switch 410 in the first position is configured to provide an electrical connection between the generator 110 through the first output connector with the first electrode 1210 and the second electrode 1220 of theArnold & Porter Ref. P35370WO00 applicator and provide an electrical connection between the generator through the second output connector with the dispersive pad 300.

[0269] Embodiment 88. The device of any one of embodiments 84 to 87, wherein the first electrode 1210 and the second electrode 1220 are in electrical connection with each other.

[0270] Embodiment 89. The device of any one of embodiments 84 to 88, wherein the dispersive pad 300 is configured to be in contact with the patient when the electric field is applied.

[0271] Embodiment 90. The device of any one of embodiments 84 to 89, wherein the second mode is a bipolar mode that generates an electric field between the first electrode 1210 and the second electrode 1220 of the applicator.

[0272] Embodiment 91. The device of any one of embodiments 84 to 90, wherein the switch in the second position is configured to provide an electronic connection between the generator 110 through the first output connector with the first electrode 1210 and provide an electronic connection between the generator 110 through the second output connector with the second electrode 1220.

[0273] Embodiment 92. The device of any one of embodiments 84 to 91, wherein the electric field of the second mode through the applicator is capable of forming a treatment zone comprising the one or more lesions.

[0274] Embodiment 93. A device for non-thermal treatment of one or more lesions in a subject in need thereof comprising an applicator 1200 comprising: a proximal end 1201; a shaft 1203; applicator tip 1205 comprising one or more first electrode 1210 and one or more second electrode 1220; and distal end 1209.

[0275] Embodiment 94. The device of any one of embodiments 93, further comprising two or more of the first electrode 1210 and two or more of the second electrode 1220.

[0276] Embodiment 95. The device of any one of embodiments 93 or 94, wherein the two or more of the first electrode 1210 and two or more of the second electrode 1220 are separated by insulating layer 1290.

[0277] Embodiment 96. The device of any one of embodiments 93 to 95, wherein the one or more first electrode 1210 and the one or more second electrode 1220 extend axially and are separated by a distance in length 1230w.

[0278] Embodiment 97. The device of any one of embodiments 93 to 96, wherein the distance in length 1230w is between 1.5 mm and 2.5 mm.

[0279] Embodiment 98. The device of any one of embodiments 93 to 97, wherein the applicator tip 1205 further comprises an opening 1207.

[0280] Embodiment 99. The device of any one of embodiments 93 to 98, wherein an opening diameter 1226 of the opening 1207 is between 1 mm and 1.5 mm.Arnold & Porter Ref. P35370WO00

[0281] Embodiment 100. A method of non-thermal treatment of a lesion in a subject in need thereof comprising: determining an electric field useful in the treatment of the lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the lesion with a device comprising: a generator comprising a first output and a second output; an clamping probe comprising a first probe and a second probe; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply the electric field of the first mode through the applicator and a second position to apply the electric field of the second mode through the applicator, wherein the electric field is within a treatment area in the subject in need thereof.

[0282] Embodiment 101. The method of embodiment 100, wherein the device further comprises a dispersive pad.

[0283] Embodiment 102. The method of any one of embodiments 100 to 101, wherein the clamping probe is clamping probe 201.

[0284] Embodiment 103. The method of any one of embodiment 100 to 102, wherein the clamping probe is clamping probe 301.

[0285] Embodiment 104. The method of any one of embodiment 100 to 103, wherein the clamping probe is clamping probe 401.

[0286] While the present disclosure has been described with reference to preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof to adapt to particular situations without departing from the scope of the present disclosure. Therefore, it is intended that the present disclosure not be limited to the particular embodiments disclosed as the best mode contemplated for carrying out the present disclosure, but that the present disclosure will include all embodiments falling within the scope and spirit of the appended claims.Arnold & Porter Ref. P35370WO00 EXAMPLES EXAMPLE 1: NON-THERMAL TREATMENT SYSTEM WITH MONOPOLAR AND BIPOLAR MODE SWITCH

[0287] Electrosurgical instruments are used to thermally ablate endometriosis lesions, either in a monopolar mode or bipolar mode. The former provides a dispersive pad on the patient to complete the circuit, while the latter applies the energy between the distal electrodes. FIG.1A shows a simple circuit that is incorporated into the generator to switch the output connectors to the different modes. FIG.2B shows clamping probe on a porcine uterine horn. For this application, the generator is in a bipolar mode- each electrode connected to the two outputs of the generator. In the monopolar mode, a dispersive pad is connected to one output of the generator and the two electrodes are connected electrically. This allows the operator to use the device as a blunt tip to apply the energy. EXAMPLE 2: IRE FOR TREATING ENDOMETRIOSIS IN A PORCINE MODEL USING A CLAMPING FORCEP APPLICATOR

[0288] A live porcine model is utilized to determine the effect of Irreversible Electroporation (IRE) on the uterine horn, bowel and ureter of a live porcine model. The electrical pulses are applied with a 5 mm X 330 mm laparoscopic bipolar forceps which generates a homogeneous electric field in the tissue between the electrodes.

[0289] A 54.5 kg female domestic pig is anesthetized. The animal is induced with an intramuscular injection of 2.2 mg / kg Ketamine HCl plus 2.2 mg / kg Xylazine plus 4.4 mg / kg Telazol (tiletamine HCl and zolazepam HCl). After endotracheal intubation the animal is maintained in a surgical plane of anesthesia on isoflurane 2-3% in oxygen. The animal is placed on a respirator using 10 to 15 ml / kg tidal volume, respiratory rate of 10-25, and pulmonary pressure not exceeding 20 cm of H2O. Lactated Ringers are administered IV at a rate of 10 ml / kg / hr.

[0290] A laparotomy is performed. The uterine horns and ureters are isolated bilaterally. The treatment areas are marked before applications using 3-0 PDS or 2-0 ePTFE sutures. The forceps are applied across the tissue, between B and C, and energy applied. FIG.8.

[0291] FIG.9 shows the waveform applied to the tissue to create the nonthermal lesions. A 500 kHz, biphasic square wave waveform is sent in packets (Cycle count) with a rest period between each packet, reducing the chance of thermal damage. For this lab, two doses are compared: 4 packets (low dose) and 8 packets (high dose).

[0292] Six lesions are made in the uterine horn, bowel, and ureter with the bipolar forceps. The tissue is grasped manually with the forceps, and a rubber band is attached to the handle to apply theArnold & Porter Ref. P35370WO00 same grasping force on the jaws for each application (not shown). Each tissue type has three lesions at the low dose and three lesions at the high dose. Additionally, each tissue type has the bipolar forceps clamped for 10 seconds without energy as a control. The pig survives for 7 hours at which point it is euthanized and the lesions harvested.

[0293] FIGS.10A and 10B depict resistance data (of the IRE signal) including recorded averages of resistance at start of each dose and changes in resistance. For example, FIG.10A depicts average resistance at start of each dose. The resistance is in a range between 60 Ω and 100 Ω for all tissues. The large standard deviation of ureter averages is due to variation of tissue filling the jaws because ureter is smaller than uterine horn and bowel. FIG.10B depicts average changes in resistance for each application, with greater magnitude of change for the higher dose. Results are useful to understand how much of load is expected to the generator. Computer simulations are used to validate the measurements and recordings.

[0294] FIG.11 depicts voltage droop data including an average voltage at start and end of all applications. For example, FIG.11 shows recorded average voltage and decrease in voltage at the end of dose for all applications. A prior application shows the average voltage for the applications is 410 V and decreased by 15 volts from start to end. As shown in FIG.11, starting voltage is 710 V and decreases by approximately 30 V. While decrease is slightly greater, the average voltage for each application is nearly 2 times the prior applications and results in more complete lesions.

[0295] Computer Simulations

[0296] A computer simulation is performed to the recorded current and to simulate the electric field zone in the tissue for ureter applications. FIGS.12A and 12B depict geometry used in computer simulation to match a recorded electric current and to simulate an electric field zone in tissues for ureter applications 16 and 19. The computer simulation is repeated until the simulated magnitudes match the recorded magnitudes.

[0297] FIG.13 depicts comparison of recorded and simulated IRE currents. FIG.13 plots recorded and simulated current for start and end of applications 16 and 19. Because cell membranes are made more permeable by the electric field, more ions flow through the cell membranes as well as around it. The value for electrical conductivity of tissue is set as a variable in the simulation: ^^^^ ^^^^ ^^^^ ^^^^^^^^ ^^^^ ^^^^ ^^^^ ^^^^=�( ^^^^ ^^^^ ^^^^ ^^^^ ^^^^ ^^^^ ∗ ^^^^ ^^^^ ^^^^) ∗ ^^^^ ^^^^ ^^^^ ^^^^_ ^^^^ ^^^^� + ^^^^ ^^^^ ^^^^ ^^^^_ ^^^^ ^^^^where factor is a dimensionless multiplier, grp is the number of packets, and esig_UH is the untreated conductivity of the uterine horn. For each tissue type, esig_UH and Factor are varied until the simulated current matched with the recorded current. The recorded current is plotted with the simulated current calculated with esiggroupset as theArnold & Porter Ref. P35370WO00 conductivity of tissues for two of the uterine horn applications (FIG. 13). For those applications, esig_UH and Factor do not change (0.28 S / m and 0.014).

[0298] The electric field strength zones are calculated in the same simulation. FIG.14 depicts simulation data at edges of treatment zone based on electric field magnitude. As shown in FIG.14, electric field may be 200 V / cm, 800 V / cm, and 1,400 V / cm at outer edges.

[0299] Histology:

[0300] FIG.15 depicts low-dose bowel applications from pathology report, showing congestion along the serosal surface, with pallor of the tunica muscularis (interpreted as acute necrosis) within the clamped region. FIG.16 depicts high-dose bowel applications from pathology report, showing loss of staining from the tunica muscularis, interpreted as acute coagulation necrosis, is present in the upper tissue. The focal loss of epithelium may be artifactual, given the lack of other associated lesions. The lower piece of tissue lacks necrosis of the tunica muscularis but has mild to moderate congestion of the submucosal vessels and moderate submucosal edema. Briefly, summary of the report follows:

[0301] Dose dependent tissue response is apparent, with clamped control tissues comprising minimal to mild changes including compression of tissues, scant hemorrhage, and endometrial or epithelial erosion. Low dose treatment elicited mild to moderate hemorrhage in some samples, with mild early necrosis (lack of staining) of muscularis layers and serosal surfaces. High-dose treatment produced greater amounts of hemorrhage and fibrin accumulation, acute necrosis with pallor of the muscularis and serosal tissues, and moderate edema of the submucosa within the bowel section. Changes are most apparent in the bowel and ureteral samples, with more mild changes in the uterine samples that were not significantly different between the low and high-dose treatments.

[0302] FIG.17 overlays simulated edges of 200 V / cm, 800 V / cm and 1,400 V / cm on MT stained lesion. The transition zone falls between 200 V / cm and 800 V / cm. This falls in a range that is expected for size of the cells that are being treated by IRE.

[0303] This study compares early treatment effects of low and high dose IRE on porcine uterine horn, bowel and uterus with a clamping bipolar device. The change in IRE resistance magnitude from group to group is substantial and may be useful in determining a complete treatment.

[0304] A dose dependent response is reported by a pathologist, and the high dose has a more fibrin accumulation for bowel and ureteral lesions. The changes are not as apparent in uterine horn lesions. The edge of treatment aligns with simulated electric field strengths in a range of expectation.Arnold & Porter Ref. P35370WO00

[0305] It is difficult to assess long term effects of IRE on these tissues based on an acute study. For the next lab, non-clamping devices are used to apply the dose. Liver can also be used as a different tissue to observe dose dependence. EXAMPLE 3: CATHETER APPLICATOR AND THERAPEUTIC SIGNATURE

[0306] The therapeutic signature of articulating tip applicator 202 is determined. Briefly, therapeutic signature may be defined as a change in electrical properties of tissues after IRE application. For radio frequency (RF) electrosurgical applications, impedance of tissues change as ions are desiccated from the heated tissue. In such case, cell membranes act as insulators to prevent ionic flow through the cell, which is be more relevant to IRE than RF electrosurgery.

[0307] Live tissue has a significant amount of capacitance in part due to cell membranes. During IRE application, cell membranes become permeable to the ions and change the impedance (both magnitude and phase). The change in magnitude and phase represent changing electrical properties of the therapeutic signature. A change in capacitance affects phase of the impedance and is dependent on the frequency of the applied signature signal (typically a sinusoidal wave is applied). To capture the therapeutic signature, the frequency is swept over a range of between 10 kHz and 1,000 kHz.

[0308] The magnitude and phase of the tissue are measured and recorded using a bio impedance board such as model AD5940 of Analog Device. The first and second electrodes are in either bipolar or monopolar mode for the IRE application; however in bipolar mode for the therapeutic signature recording. During the IRE application, magnitude and phase are determined based on a real-time feedback control member, and the generator indicates a complete treatment based on the therapeutic signature.

[0309] FIGS.18A and 18B depicts the experimental apparatus for determining the therapeutic signature in a potato. The experimental apparatus comprises articulating tip applicator 202 as the applicator, as shown in FIGS.3A, 3B, 4A, and 4B. FIG.18B depicts various results from treatment zones (e.g., 401, 404, 405, 408) 24 hours after 8 applications. The initial recordings are made with an IRE dose defined in Table 1 while the voltage is varied. The IRE dose is determined experimentally in live porcine tissue. Table 1Arnold & Porter Ref. P35370WO00

[0310] FIGS.19A and 19B depict an example of a non-thermal treatment system, according to one or more aspects described herein. As depicted in FIG.19B, a small red potato is sliced and articulating tip applicator 202 is moved forward until second electrode 220 contacts the potato. Before and after each bipolar IRE application, the AD5940 measures and records magnitude and phase with respect to a frequency in a range between 10 kHz and 200 kHz. IRE is applied at 500 V (low dose), 700 V and 1 kV (high dose) while voltage and current are recorded with a Picoscope.

[0311] FIG.20 and FIG.21 depict magnitude and phase of the potato before and after IRE treatments with respect to a frequency in a range of 10 kHz to 200 kHz, respectively. The magnitude decreases and phase moves more positive after the treatment indicating that conductivity and capacitance decrease when cell membranes (walls for potatoes) are porated or damaged. Table 2

[0312] FIGS.22 and 23 depict linear regression plots to quantify the change. Table 2 discloses a comparison with a difference in slope of the magnitude and phase. While both magnitude and voltage changes after an IRE treatment, it is noted that phase changes significantly more.

[0313] FIG.24 depicts a plot of magnitude and phase with the catheter in tap water. As a control, catheter may be placed in tap water, which is conductive and contains molecules but no cell structures. It is noted that there is little to no change in magnitude and phase after IRE treatment of the tap water.

[0314] It is well known that impedance of tissue decreases when heated by RF electrosurgery and then rapidly increases as the tissue is desiccated. IRE application does not significantly increase temperature of tissue. Therefore, one may not expect similar changes in impedance as with RF. However, based on various experimental results, there is a linear change in both magnitude and phase.

[0315] Zhao et. al report on and use of impedance spectroscopy with a multi-electrode IRE ablation system. Zhao, Y., et al., “Ablation outcome of irreversible electroporation on potato monitored by impedance spectrum under multi-electrode system.” Biomedical Engineering Online, 126 (2018). A monophasic IRE waveform is applied to potato tubers with active electrodes, whileArnold & Porter Ref. P35370WO00 the inactive electrodes were used to record the impedance with an impedance analyzer. The frequency is in a range between 1 Hz and 1 MHz, while the IRE generator applied a signal with varying amplitude, pulse width and pulse number. An equivalent circuit model (ECM) is used to “extract” more information from impedance data as stated by the author. The impedance is defined as:where cpe is a constant phase element, ω is the angular frequency, j is the unit imaginary number, Q and n are the empirical parameters. Zhao et al. report that the polarization of the cell membrane at a tissue level is described by another CPE while in cell level it is equivalent to a capacitor.

[0316] FIG.25A depicts Cole-Cole equivalent circuit model and FIG.25B depicts magnitude and phase of the Impedance as reported by Zhao, et al.

[0317] According to Zhao et al., terms Rcand Reare defined as resistances for intracellular and extracellular current pathways as part of thew Cole-Cole equivalent circuit model as shown in FIG. 25A. It is not entirely clear how these (presumably) resistance magnitudes are part of Zcpe.

[0318] FIG.25B depicts impedance and phase results from Zhao et al. See Zhao, et al. at figure 3a. Both magnitude and phase are highly non-linear with respect to frequency in a range between 1 Hz and 1MHz, before and after electroporation.

[0319] Contrary to the results of Zhao, et al., the results presented in the present disclosure show a linear response with respect to frequency in a range between 10 kHz and 200 kHz when biphasic IRE waveform is applied to the potato using a two-electrode system (the same electrodes are used to apply the energy and record magnitude and phase). As depicted in Zhao, et al., Zhao’s magnitude may be interpreted as a linear change, and phase data may be interpreted as highly non- linear change. FIG.25B, in a range of a box 2502. EXAMPLE 4 – PROCESS FOR DETERMINING IF THE IRE APPLICATION IS COMPLETE

[0320] FIG.26 illustrates an example of a process 2600 for testing a non-thermal treatment system, according to one or more aspects described herein. The operations of process 2600 presented below are intended to be illustrative and, as such, should not be viewed as limiting. In some implementations, process 2600 may be accomplished with one or more additional operations not described, and / or without one or more of the operations discussed. In some implementations, two or more of the operations of process 2600 may occur substantially simultaneously. The described operations may be accomplished using some or all of the components described in detail above with respect to non-thermal treatment system 100.Arnold & Porter Ref. P35370WO00

[0321] In an operation 2602, process 2600 may include determining an electric field useful in a treatment to form a determined electric field. For example, IRE dose such as cycle count, rest period(s), group number, dead time(s) (shown in Table.1) may be determined as previously described herein. In an operation 2604, process 2600 may include applying the determined electric field of a first mode or an electric field of a second mode to a lesion. In an operation 2606, process 2600 may include determining a difference between the first signal and the second signal. In an operation 2608, process 2600 may include comparing (or determining) whether the difference exceeds a predetermined threshold.

[0322] In an operation 2610, process 2600 may include instructing the switch to change to a desired position if the difference exceeds the predetermined threshold of the non-thermal treatment system 100. In some embodiments, process 2600 may instruct the application to end if the difference exceeds the predetermined threshold. In other embodiments, process 2600 may instruct the application to send an alert to operator to end the operation. The alert may be, for example, visual, audible, tactile (vibration), other means of sending messages or a combination thereof. Then, the operator may end the operation using switch such as foot switch. However, the operator may continue the operation based on the feedback information from the sensors.

[0323] In an operation 2612, process 2600 may include instructing the switch to maintain the current position if the difference does not exceed the predetermined threshold. In some embodiments, process 2600 may instruct the application to maintain current status if the difference exceeds the predetermined threshold. In other embodiments, process 2600 may instruct the application to send an alert to operator regarding the current status. The alert may be, for example, visual, audible, tactile (vibration), other means of sending messages or a combination thereof. However, the operator may end the operation based on the feedback information from the sensors.

[0324] During process 2600, at least operations 2606 and 2608 may be repeated until the difference between the first signal and the second signal is within the predetermined threshold. EXAMPLE 6 : PERITONEUM AND BOWEL SIGNATURE

[0325] A study is performed to determine the change in the electrical signature of live porcine peritoneum and bowel tissue after treatment with IRE. Briefly, a 54.5 kg female domestic pig is anesthetized. The animal is induced with an intramuscular injection of 2.2 mg / kg Ketamine HCl plus 2.2 mg / kg Xylazine plus 4.4 mg / kg Telazol (tiletamine HCl and zolazepam HCl). After endotracheal intubation the animal is maintained in a surgical plane of anesthesia on isoflurane 2- 3% in oxygen. The animal is placed on a respirator using 10 to 15 ml / kg tidal volume, respiratoryArnold & Porter Ref. P35370WO00 rate of 10-25, and pulmonary pressure not exceeding 20 cm of H2O. Lactated Ringers are administered intravenously at a rate of 10 ml / kg / hr.

[0326] A laparotomy is performed. IRE applications are performed on a liver, small intestine and parietal peritoneum of the abdominal wall. Electrical response are characterized for each application. Approximately 6.0 hours after the last IRE application, the animal is sacrificed, and all the application sites are harvested.

[0327] FIG.27 depicts applicators comprising varied distance in length (230w) along with the tissues treated. The applicator contain concentric bipolar electrodes with different inner (first) electrode diameters. For example the peritoneum (FIG.27C) is treated with an articulating tip applicator 202 (i.e., Applicator 06Sep2023A) comprising an inner electrode (also referred to as a first electrode) with a 3 mm diameter and an outer electrode (also referred to as the second electrode) with a 6 mm diameter (FIG.27A). The bowel (FIG.27D) is treated with an applicator 202 (i.e., Applicator 08Sep2023A comprising an inner (first) electrode with a 2 mm diameter and an outer (second) electrode comprising a 6 mm diameter (small electrode, FIG.27B). A total of 18 applications are made (6 on the peritoneum using Articulating tip applicator 202 (06Sep2023A), 12 on the jejunum using Articulating tip applicator 202 (06Sep2023A) and articulating tip applicator 202 (08Sep2023A). Table 3 defines the three doses (low, medium and high). Each group contains 10 cycles of 500 kHz (2 µs period, 1 µs pulse width). Table 3

[0328] The AD5940 application board (Analog Instruments) are used to measure and record magnitude and phase of the tissue over a range of sinusoid waveforms (10 kHz to 180 kHz in increments of 18 kHz). The IRE waveform is recorded with an oscilloscope (PicoScope), a high differential voltage probe (HDVP) and a current transformer (Pearson). A switching box is used to switch each applicator between the AD5940 for a pre-signature, the IRE generator for a therapeutic application, and back to the AD5940 for a post signature recording.

[0329] FIG.28 depicts various magnitude plots of the larger electrode applicator, Articulating tip applicator 202 (06Sep2023A) on peritoneum, and FIG.29 depicts various phase plots of Articulating tip applicator 202 (06Sep2023A) on peritoneum. The solid lines are before and dashed lines indicate after IRE application. More specifically, FIGS.28-29 plot magnitude and phase ofArnold & Porter Ref. P35370WO00 Articulating tip applicator 202 (06Sep2023A) on peritoneum before and after the three doses (low- blue, medium- red, and high- green). Over the range of frequencies, magnitudes decreases and phase becomes more positive after IRE treatment, as is also seen in potatoes (Example 6, FIGS. 28-29). Changes of magnitude and phase in sinusoidal waveform is an indication that electrical conductivity is increased (magnitude) and capacitance has decreased (phase), consistent with cell electroporation.

[0330] FIG.30 depicts magnitude plots of Articulating tip applicator 202 on jejunum, and FIG. 31 depicts phase plots of articulating tip applicator 202 (06Sep2023A) on jejunum, before (solid lines) and after (dashed lines) IRE application. FIGS.30-31 show similar trends in the jejunum as shown with results of peritoneum.

[0331] FIG.32 depicts various magnitude plots of treatment with the smaller inner electrode, Articulating tip applicator 202, on jejunum, and FIG.33 depicts various phase plots of Articulating tip applicator 202 on jejunum, before (solid lines) and after (dashed lines) IRE application. While both magnitudes of treatment with the large inner electrode (Articulating tip applicator 202) and the smaller inner electrode (Articulating tip applicator 202) decrease after IRE treatment on the jejunum, treatment with the larger first electrode (articulating tip applicator 202) results in a higher initial magnitude, likely due to increased total electrode surface area of the smaller inner electrode (articulating tip applicator 202) which comprises a longer electrode. When comparing change in phase after IRE treatment for both electrode applications on jejunum, treatment with the smaller electrode becomes positive for 20 groups and 30 groups at 80 kHz and higher but treatment with the larger electrode remains negative.

[0332] Review of tissue after the treatment dose with medium (app 15 and app 16) and high (app 17 and app 18) dose applications of smaller diameter inner electrode on jejunum shows some treatment zones with white spots (FIG.34). White zones of apps 16, 17 and 18 may be due to heating of the tissue. The 2 mm diameter electrode produces a very high current density. While images of the other peritoneum and jejunum post treatments are not recorded, white zones are observed only on apps 16-18. The same applications are repeated on postmortem porcine uterine horn (See below, FIG.42A and B), and there is no evidence of thermal damage (white zones), or positive phase angle (FIG.44). White zones are not expected to be seen in postmortem tissue due to contractions but thermal damage can be expected on postmortem tissue.

[0333] IRE Measurement Results

[0334] The IRE waveform is calculated by dividing the voltage and the current magnitude at the same point on the peak of the waveform. Based on the trends in the change of IRE magnitudeArnold & Porter Ref. P35370WO00 shown in FIG.35, there appears to be no significant difference between low, medium, and high doses.

[0335] Signature Analysis

[0336] FIGS.36 through 41, which plot the initial magnitude and phase and the magnitude and phase after IRE treatment over 10 kHz to 180 kHz, illustrate that the electrical tissue properties (the signature) may change after IRE treatment. Reviewing the change at the 10 discrete frequencies recorded, 10 kHz to 180 kHz in 18 kHz increments show that the peritoneum has a significant difference of magnitude at 10 kHz and 180 kHz, while differences in jejunum for the larger and smaller electrode are less but similar values and trends, possibly indicating tissue type dependence of the signature change (FIGS.36, 37, and 38). It may be for each plot, magnitude change after IRE treatment with 10 kHz is greater than the change with 180 kHz. The opposite of magnitude is observed for phase – the phase change is greater at 180 kHz than at 10 kHz. (FIGS.39, 40, and 41). FIGS.39, 40, and 41 plot phase difference after IRE treatment for large electrode peritoneum, large electrode jejunum and small electrode jejunum, respectively, at each frequency with the change in phase at 10 kHz and 180 kHz inserted. The numbers for each magnitude and phase at 10 kHz and 180 kHz are shown in Table 4. Table 4

[0337] The complex impedance is defined as: ^^^^ = ^^^^ − ^^^^ ^^^^ ^^^^ = ^^^^ ^^^^ ^^^^−1(^^^^) ^^^^ where R is real part of the impedance and X is imaginary part or the reactance. Θ is phase angle (positive for inductance and negative for capacitance). In a circuit, the real part is represented byArnold & Porter Ref. P35370WO00 resistors, and the imaginary part is represented by capacitors and / or inductors. In tissue, the resistance may be calculated from the complex conductivity (Miklavcic 2006): ^^^^∗= ( ^^^^ + ^^^^ ^^^^ ^^^^) where σ is conductivity, ω is angular frequency and ε is permittivity. The real part is ionic flow, and the imaginary part is defined mostly be capacitance of the cell membranes. There are more complicated ways to define tissue impedance, but that is not the purpose here. The magnitude and phase of peritoneum and jejunum have been measured and presented here before and after IRE treatment.

[0338] The tissue signature is defined here as resistive magnitude of IRE application and magnitude and phase of a range of low power sinusoidal signals (10 kHz to 180 kHz), or a combination. The change of the signature is presented here to determine whether low or high dose has been applied, because the low dose is not a complete treatment while the high dose is a complete treatment. In various situations, operator may apply various dose, e.g., single, low or high dose. Since change in IRE magnitude is inconclusive, a combination of sinusoidal magnitude and phase may be recommended. Table 4 shows that magnitude changes the most at 10 kHz and phase changes the most at 180 kHz. It is therefore recommended that a range of magnitude change at 10 kHz and a range of change at 180 kHz be defined as the signature change.

[0339] It appears that there is no significant difference between the electrode designs on the bowel (jejunum). It may be advantageous that signature change is not dependent on electrode design. However, there does appear to be an association with tissue type.

[0340] The definition of the signature change to determine high or low dose is a range of magnitude change at 10 kHz and a range of phase change at 180 kHz.

[0341] Postmortem porcine uterine horn

[0342] The experiments are repeated on postmortem porcine uterine horn, as described above to determine source of white zones.

[0343] FIGS.42A and 42B show postmortem porcine uterine horn (26° C) after treatment with the smaller electrode at 30 groups. It lacks white zone shown in FIG.34, with the same probe and application in live porcine jejunum. If the white tissue in Figure 2 is due to thermal damage, the same should be observed in Figure 8.

[0344] FIGS.43-44 plot magnitude and phase of the application shown in FIGS.42A and 42B. The magnitude curves follow the similar pattern as shown in FIG.32 (data of FIG.43 at a higher magnitude). The phase plots (FIG.44) show an increase in phase angle after IRE treatment (solid to dotted lines) but is more negative at the higher sinusoidal frequencies. This may be an indication of dead tissue.Arnold & Porter Ref. P35370WO00 EXAMPLE 7 : URETER IRE SIGNATURE

[0345] This study determines change in electrical signature of ureter after applying and IRE dose with a bipolar clamp probe. The “electrical signature” is based on a resistive magnitude of IRE application and the magnitude and phase of a range of low power sinusoidal signals (10 kHz to 180 kHz). The signature may be a combination of all three measured before and after the IRE treatment.

[0346] A 54.5 kg female domestic pig is anesthetized. The animal is induced with an intramuscular injection of 2.2 mg / kg Ketamine HCl plus 2.2 mg / kg Xylazine plus 4.4 mg / kg Telazol (tiletamine HCl and zolazepam HCl). After endotracheal intubation the animal is maintained in a surgical plane of anesthesia on isoflurane 2-3% in oxygen. The animal is placed on a respirator using 10 to 15 ml / kg tidal volume, respiratory rate of 10-25, and pulmonary pressure not exceeding 20 cm of H2O. Lactated Ringers are administered intravenously at a rate of 10 ml / kg / hr.

[0347] A laparotomy is performed. IRE applications are performed on a liver, small intestine and parietal peritoneum of the abdominal wall. Electrical response is characterized for each application. Approximately 6.0 hours after the last IRE application, the animal is sacrificed, and all the application sites are harvested.

[0348] The entire application procedure is recorded with a video camera.

[0349] FIG.51 depicts two devices tested: (1) a bipolar articulating tip applicator 202 used on peritoneum and jejunum and (2) the bipolar clamp probe on the ureter. A total of 6 applications are made on the ureter (low, medium, and high and a repeat of each) at 800 V. Table 5 describes the three doses. Table 5

[0350] The AD5940 application board (Analog Instruments) is used to measure and record magnitude and phase of a tissue over a range of sinusoid waveforms (10 kHz to 180 kHz in increments of 18 kHz). The IRE waveform is recorded with an oscilloscope (PicoScope series 6000), a high differential voltage probe (HDVP) and a current transformer (Pearson Electronics). A switching box is used to switch each applicator between the AD5940 for a pre-signature, the IRE generator for a therapeutic application, and back to the AD5940 for a post signature recording.Arnold & Porter Ref. P35370WO00

[0351] Sinusoidal Results

[0352] FIGS.52-53 plot magnitude and phase of the applications on the ureter. The magnitudes decrease and the phase becomes more positive after IRE treatment, as previously observed in potatoes. The change of magnitude and phase of the sinusoidal waveform is an indication that electrical conductivity has increased (magnitude) and capacitance has decreased (phase), consistent with cell electroporation.

[0353] The phase of clamped tissue is positive over the range of 90 kHz to 180 kHz before and after treatment (FIG.53). The shifting of the phase in the positive direction after treatment is observed, with a greater increase. The positive phase is observed when the bipolar device is used on jejunum. It is not immediately clear how the tissue could have a positive phase shift. The AD5940 operates in two output modes; 2-wire and 4-wire. For these recordings, a 2-wire is used, and the positive signal could be inductance in the system and not the tissue. The recordings are repeated with the 4-wire output.

[0354] Similar trends are observed in postmortem porcine uterine horn with the Clamp FIGS. 59-60.

[0355] IRE Measurement Results FIG.54 is a graph showing a plot of calculated resistances with the technique shown on the inset image. The IRE waveform is calculated by dividing the voltage and the current magnitude at the same point on the peak of the waveform.

[0356] There appears to be a clear trend of decreasing IRE resistance magnitude with the number of groups. This is not observed when the bipolar probe is used to apply IRE doses on the peritoneum and jejunum. This may be attributed to higher intensity electric field between clamp electrodes as compared with the peritoneal tissue.

[0357] FIG.55 compares simulation of an electric field over a range of 800 V / cm (blue) to 6 kV / cm (red) for the bipolar applicator 202 and the bipolar clamping probe. Both plots are set to display a minimum of 800 V / cm (blue), the lowest value at the edge of the tissue for the clamp probe is about ≈ 3kV / cm (green). Therefore, entire volume of the tissue in the clamp probe is exposed to a higher field strength, leading to more tissue effect and subsequent change in the IRE magnitude.

[0358] Signature Analysis

[0359] FIGS.56-57 plot magnitude and phase difference after IRE treatment for clamped ureter at each frequency. The change in magnitude is similar to the large electrode jejunum with the bipolar device. There is a significant difference between the low dose and high dose for the phaseArnold & Porter Ref. P35370WO00 recordings, but for the bipolar applications the high dose produced a positive slope (not shown), whereas the slope of the change after the high dose is negative for the clamped ureter.

[0360] In order to determine change in the electrical signature of ureter after applying and IRE dose with bipolar clamp probe. During this same lab, bipolar device is used on peritoneum and jejunum. While there are several similarities between the two devices on live tissue, there are two clear differences. First, the bipolar clamp probe on the ureter had a correlation of the IRE magnitude with the number of groups. This is not as strong correlation with the bipolar applicator 202. Second, the slope of the difference in phase before and after IRE treatment is negative for the forceps applications and positive for the clamp applications. In various cases, a surgeon chooses either instrument (e.g., forceps or clamp) depending on a size and location of an endometrial lesion.

[0361] With the peritoneum and jejunum applications, it is concluded that the definition of the signature change to determine a high dose or low dose is a range of magnitude change at 10 kHz and a range of phase change at 180 kHz. Additional data is presented here for a different applicator. It appears that a change in IRE magnitude is a strong indication that the clamp application is complete. It is important to determine if the AD5940 board measured and recorded positive phase angles in 4-wire output configuration.

[0362] FIGS.58-59 show the clamp on post mortem porcine uterine horn applying 800 V. No thermal damage is observed.

[0363] FIGS.60-61 plot magnitude and phase of the applications on postmortem uterine horn clamped.

[0364] EXAMPLE 8: A COMPARISON OF THE SIGNATURES OF LIVE AND POSTMORTEM TISSUE Signature recordings from a lab (in-vivo jejunum recordings) is compared with thawed postmortem porcine jejunum. The generator with the AD4950 board is rebuilt for postmortem experiments. FIGS.27A, 27B, 27C, and 27D and Table 6 show devices and settings from a lab (Example 6). The results with the 3 mm diameter inner electrode are compared with the postmortem results. A switchbox is used to switch input to the probe between the AD4950 board and the IRE therapeutic dose. The bipolar forceps are used on the live ureter and are not be used for comparison. Table 6Arnold & Porter Ref. P35370WO00

[0365] Postmortem Methods

[0366] FIG.62 shows articulating tip applicator 202 on a section of postmortem small bowel. Table 7 shows settings for the IRE dose, with a set voltage of 800 volts, and the groups are 10 and 30 (same as the live tissue applications on small bowel). Table 7

[0367] The generator (POC 002) has an AD4950 board built in so that initial signal measurement (magnitude and phase over a range of sinusoidal frequencies) of small bowel may be recorded, followed by an IRE dose, and a final signature measurement of the post treated small bowel without using a switchbox.

[0368] Live Tissue Results

[0369] FIG.63 is a plot of the mean (n=2) signature magnitude of 10 group treatments, as well as a linear curve fit of the data. The slopes of before and after curves are -45.9 e-5 and -40.1 e-5, show little change.

[0370] FIG.64 is a plot of the mean (n=2) signature magnitude of 30 group treatments, as well as a linear curve fit of the data. The slopes of before and after curves are -42.1 e-5 and -21.9 e-5, respectively, a larger change than with 10 groups.

[0371] FIG.65 is a plot of the mean (n=2) signature phase of 10 group treatments, as well as a linear curve fit of the data. The slopes of before and after curves show little change.

[0372] FIG.66 is a plot of the mean (n=2) signature phase of 30 group treatments, as well as a linear curve fit of the data. The slopes of before and after curves change from negative slope to positive slope.

[0373] Postmortem Tissue Results

[0374] FIG.67 is a plot of the mean (n=3) signature magnitude of 10 group treatments, as well as a linear curve fit of the data. The slopes of before and after show a small change.

[0375] FIG.68 is a plot of the mean (n=3) signature magnitude of 30 group treatments, as well as a linear curve fit of the data. The slopes of before and after curves are -16.5 e-5 and -10.3 e-5, respectively, a larger change than with 10 groups.Arnold & Porter Ref. P35370WO00

[0376] FIG.69 is a plot of the mean (n=3) signature phase of 10 group treatments, as well as a linear curve fit of the data. The slopes of before and after curves are -1.01 e-5 and -1.36 e-5, respectively-a small change.

[0377] FIG.70 is a plot of the mean (n=3) signature phase of 30 group treatments, as well as a linear curve fit of the data. The slopes of before and after curves are a large change from negative slope to positive slope.

[0378] FIG.71 compares the slopes of the linear curves fit to each set of data. The magnitude of slope change is much greater for live tissue compared to postmortem tissue, while the phase is about the same between the two tissues. There is a significant difference between 10 group (“Diff10G”; low dose) and 30 group (“Diff30G”; high dose) for both magnitude and phase of both tissues.

[0379] This study compares the signature between postmortem and live porcine jejunum before and after a low IRE dose and a high IRE dose. The postmortem jejunum is a reasonable tissue phantom when looking at the change in the slope of the phase over a range of 10 kHz to 180 kHz. The magnitude trends positive in a fairly linear fashion, whereas the phase actually transitions from negative to positive after a high IRE dose (FIG.70) but stays negative after a low IRE dose (FIG. 69). While a quadratic curve fit may be more accurate for phase, one could also look at the slope over the more linear range (40 kHz to 180 kHz). The change in the slope of the phase alone may be the most accurate indicator (“signature”) of a complete dose. EXAMPLE 9: FURTHER SIGNATURE ANALYSIS OF POSTMORTEM TISSUE

[0380] Additional signature analysis is performed on postmortem tissue. An experimental setup to record the sinusoidal signal before and after the IRE treatment is utilized as shown in FIG.72. The AD4950 board is built into the generator, which automatically switches between recording the sinusoidal signal and the IRE treatment voltage and current. The camera records a close-up image of the contact between the distal end of the probe and the small bowel.

[0381] Table 8 lists variables for the applications. All applications have a cycle count of 10, a rest period of 0.005 seconds and a dead time of 0.1 seconds. Table 8Arnold & Porter Ref. P35370WO00

[0382] FIG.73 provides two applications with different electrode contact. FIGS.77-79 describe in more detail how electrode contact area of sinusoidal magnitude changes.

[0383] Table 9 lists the settings for Group 1. Articulating tip applicator 202 (i.e., Applicator 27Sep2023A using applicator settings used in FIGS.27A, 27B, 27C, and 27D, but based on settings listed in Table 9. The applicator contain concentric bipolar electrodes with different inner (first) electrode diameters. For example the peritoneum (FIG.27C) is treated with an articulating tip applicator 202 (i.e., Applicator 06Sep2023A) comprising an inner electrode (also referred to as a first electrode) with a 3 mm diameter and an outer electrode (also referred to as the second electrode) with a 6 mm diameter (FIG.27A). The bowel (FIG.27D) is treated with an articulating tip applicator 202 (i.e., Applicator 08Sep2023A comprising an inner (first) electrode with a 2 mm diameter and an outer (second) electrode comprising a 6 mm diameter (small electrode, FIG. 27B).) is used for all applications, and a total of three applications (N=3) is measured and recorded for each setting. Table 9

[0384] FIG.74 is a plot of mean phase (and fitted regression line) of Group 1A and Group 3A (see Table 8) to show the difference when more total pulses are applied. For each application, the difference in the slope of an equation in FIG.74 is calculated. These differences are then compared to determine the effect of packet count, group number and total pulses.

[0385] FIG.75 shows the difference of the slope of the phase for each application after IRE treatment. The X axis labels have been changed for easier comparison (e.g. Group 1A is now G1- 5P10G where 5P indicates for 5 packets and 10G indications 10 groups). Groups 1 and 2 show how the total number of pulses increases the change in the slope. The delay between the packets is a rest period (0.005 s) and a delay between the groups is a dead time (0.1 s), therefore more packets and fewer groups to apply the same number of pulses would be a faster application. FIG.75 showsArnold & Porter Ref. P35370WO00 that change in the slope is dependent on the total number of pulses and not a combination of packets and groups.

[0386] For example, Group 4 has the smallest change in the slope. This may be due to the excessive amount of energy that leads to coagulation of the proteins and thus the load becomes more homogenous and less reactive (cells in the extracellular matrix are destroyed, not just electroporated). As an example, the black arrows in FIG.76B point to areas of tissue which have been coagulated.

[0387] A range of 50 kHz to 180 kHz produces a more linear change in the slope of the phase than a range of 10 kHz to 180 kHz (see FIG.77).

[0388] A greater number of pulses correlates with a larger change in the slope, until excessive energy is applied to damage the structure of the tissue. More packets and fewer groups produce the same change in slope as fewer packets and more groups. More packets add little to the total application time, whereas more groups can increase it substantially.

[0389] The change in the slope of the phase over 50 kHz to 180 kHz is the best indicator of tissue treatment after IRE application. What remains is to correlate the depth of treatment in live tissue to these slope changes—a greater slope change correlates with a deeper treatment.

[0390] FIG.77 plots the phase of 10 groups over the frequency range of 10 kHz to 180 kHz. FIG.74 plots the phase over a range of 50 kHz to 180 kHz—resulting in a much better curve fit.

[0391] The magnitude of the sinusoidal signal is used here to determine tissue contact with the electrodes. FIGS.78-79 plot the mean of the magnitude before an application of IRE. The difference in group 1 magnitude is small (≈25 Ω) whereas the difference in Group 30 is larger (≈125 Ω). This is due to the outer electrode not being in contact with the tissue. EXAMPLE 10: BIPOLAR CLAMPING PROBE ON POSTMORTEM TISSUE

[0392] A bipolar clamp probe is tested on postmortem tissue. FIGS.80-81 and Table 10 depict device and settings used for postmortem porcine small bowel experiments. Table 10Arnold & Porter Ref. P35370WO00

[0393] FIGS.82-83 depict plots of fitted curves of the phase before and after IRE treatment, with a frequency range is 10 kHz to 180 kHz.

[0394] FIG.84 depicts a plot of a difference in magnitude of the slope from FIGS.82-83. As expected, magnitude increases with more IRE treatment. However, the decrease of group 4 may be due to thermal coagulation of the tissue.

[0395] FIGS.85 depicts a photograph of lesions after treatment. The thermal coagulation can be seen if the treatments with a greatest number of pulses.

[0396] The bipolar clamping device with wire jaws (shown in FIGS.80) produces a signature phase which begins negative at 10 kHz and becomes positive at 50 kHz. This is observed with the bipolar clamp device with solid jaws (see Example 7). It is not clear why a positive phase is recorded when tissue is mostly capacitive (negative phase), but there is a clear correlation between the number of pulses applied and change of slope of the phase. For articulating tip applicator 202, the frequency range is 50 kHz to 180 kHz (linear part of the curve), while for this probe, the frequency range is 10 kHz to 180 kHz (linear part of the curve). EXAMPLE 11: ADDITIONAL SIGNATURE ANALYSIS

[0397] Additional studies are performed to identify the signature analysis. The electrical signature is a definite, identifiable, measurable change in one or more physical (tissue) quantities to indicate completion of an IRE application. This Example includes 1) change in slope of phase with bipolar clamping forceps, and 2) observations of phase between voltage and current of a recorded IRE waveform.

[0398] An experimental setup is performed to record sinusoidal signal before and after the IRE treatment. FIG.86 The AD4950 board, measuring magnitude and phase is built into the generator, which automatically switches between recording sinusoidal signal and IRE treatment voltage and current. A camera records a close-up image of a contact between the distal end of a probe and a small bowel.Arnold & Porter Ref. P35370WO00

[0399] FIG.87 provides the distal end of three bipolar clamps tested as exemplary embodiments, and FIG.88 depicts images of each device clamped onto the tissue according to one embodiment of the present disclosure. For example, FIG.87 depicts three bipolar clamps tested, (a - “BiClamp5”), (b - “Maryland1”), and (c - “BiClamp2") with surface area of one jaw contact area, which affects an amount of thermal damage to tissue as the number of total pulses is increased. Examples of each device clamped onto the tissue are shown in FIG.88.

[0400] Table 11 lists settings for four (4) sets of applications for each device. The means of sinusoidal phase are calculated from 3 applications for a range of 10 kHz to 180 kHz. A comparison of change in a fitted slope of the phase is then made for four sets. Table 11

[0401] FIG.89 shows slopes of phase before and after using BiClamp5 device (400 pulses and 1,000 pulses), and FIG.90 are graphs showing slopes of the phase before and after using BiClamp2 device (2,000 pulses and 3,000 pulses). FIG.91 shows the difference for each set of plots in FIGS.89-90. All sets are 10 groups.

[0402] FIGS.89-90 plot the BiClamp2 slope of the phase before and after the IRE treatment for sets 1 and 2, and 3 and 4, respectively, while FIG.91 plots differences in slope for each set. As reported, phase becomes positive at about 50 kHz when a clamp probe is used. FIG.91 depicts an increase in the slope as number of packets increases.

[0403] FIG.92 are graphs showing slopes of phase before and after using Maryland1 device (400 pulses and 1,000 pulses), and FIG.93 are graphs showing slopes of the phase before and after using Maryland1 device (2,000 pulses and 3,000 pulses). FIG.94 is a graph showing difference for each set of plots in FIGS.92-93. All sets include 10 groups.

[0404] FIGS.92-93 plot Maryland1 slope of the phase before and after the IRE treatment for sets A and B, and C and D, respectively, while FIG.94 plots the difference in the slope for each set. The increase in the difference of the slopes are shown in FIG.94 from 400 pulses to 1,000 pulses but decreases at 2,000 and 3,000 pulses. This may be due to excessive thermal damage by the Maryland1 which has a smaller surface area and produces a higher current density (FIG.95).

[0405] FIG.95 depicts treatment zone after 2,000 pulses for the BiClamp5 and the Maryland1.Arnold & Porter Ref. P35370WO00

[0406] FIG.96 are graphs showing slopes of phase before and after using BiClamp2 device (400 pulses and 1,000 pulses), and graphs in FIG.97 show slopes of the phase before and after using BiClamp2 device (2,000 pulses and 3,000 pulses). FIG.98 is a graph showing the difference for each set of plots in FIGS.96-97. All sets are 10 groups.

[0407] FIGS.96-97 plot the BiClamp5 slope of the phase before and after the IRE treatment for sets A and B, and C and D, respectively, while FIG.98 plots difference in slope for each set. The difference of the slopes increase but not in a linear fashion as pulses increase.

[0408] FIG.99 depicts treatment zones with thermal damage for BiClamp2 at 1,000, 2,000 and 3,000 pulses.

[0409] FIG.100 depicts phase shift between IRE voltage and IRE current for clamp probe and articulating tip applicator 202 according to one embodiment of the present disclosure.

[0410] FIG.100 plots voltage (blue) and current (red) for a clamp application and a articulating tip applicator 202 application. Phase shifts are shown for each application. The current leads the voltage in the clamp recording, which indicates a positive phase shift or an inductive load. For the articulating tip applicator 202, voltage leads the current, which indicates a negative phase shift or capacitive load. This shift in the IRE waveform is supportive of a positive phase shift with the clamp for the signature analysis.

[0411] Based on observation of the signature the slope of phase of the signature increases as pulses increase and is an indicator of more IRE treatment. Further, when thermal damage of the tissue occurs, the slope tends to decrease, which is likely an indicator of thermal damage. Moreover, the positive phase shift between IRE voltage and current for the clamp supports the same observation with a sinusoid signal. EXAMPLE 12: IRE AND SIGNATURE ON LIVE JEJUNUM

[0412] This study repeats the applications of Example 11, where BiClamp devices are tested on postmortem porcine jejunum. The report found that slope of a phase of the signature increases as pulses increase and is an indicator of greater (or over-, excessive) IRE treatment. Similar BiClamp devices are to be tested on live porcine jejunum.

[0413] FIG.101 illustrates an exemplary embodiment of various components, for example, a) live jejunum accessed through a laparotomy and b) BiClamp devices tested on the jejunum.

[0414] FIG.101 depicts an access to a porcine jejunum and devices tested, while Table 1 lists settings for a matrix of applications. The OreoSig 001 generator (not shown) produced IRE pulses and measured the signature before and after each application. The voltage and current of the IRE output are recorded with a Picoscope oscilloscope. Three applications for each Group_set areArnold & Porter Ref. P35370WO00 recorded to calculate a mean value. After each application, a photo is taken of a treatment zone. The device is moved along the jejunum until all 24 applications are completed.

[0415] Table 12 lists settings for group sets of applications for each device. All applications are 10 cycles, 0.005 sec rest period and 0.1 sec dead time. Table 12

[0416] FIG.102 plots slope of a phase of clamped tissue before and after the IRE application. A linear curve is fit to the data. Slope of the linear curve is then compared for each Group_set application. More specifically, FIG.102 plots a linear curve fitted to value of a phase over a range of 10 kHz to 180 kHz. The slope of a fitted curve is labelled near the curve.

[0417] FIG.103 illustrates the difference in a fitted line of a phase slope. More specifically, FIG.103 plots the difference of a phase slope fitted curve for four BiClamp7 applications. For 400 pulses, the difference is negative and becomes positive at 1,000 pulses, but shows a linearly increasing trend.

[0418] FIG.104 plots differences of a phase slope fitted curve for four BiClamp8 applications. For 800 pulses, the difference is negative and becomes positive at 1,700 pulses, again showing a regular increase. These device and settings are tested in postmortem porcine jejunum. While the trend of an increase in difference of a phase slope as total number of pulses increased is observed, there are observations in postmortem jejunum that are not seen here. The phase of the postmortem jejunum is positive at 50 kHz and greater for all of the before and after recordings, while the phase in the live jejunum tissue is positive only after the IRE treatment at frequencies greater than 60 to 100 kHz, likely indication a limitation to postmortem tissue as a phantom.

[0419] FIG.105 illustrates treatment zones (black arrows) for 400 and 1,000 pulses for BiClamp7 device, and FIG.106 illustrates treatment zones for 2,000 and 3,000 pulses for BiClamp8 device.Arnold & Porter Ref. P35370WO00

[0420] FIGS.105-106 are photographs of treated zones for the fewest pulses and most pulses, respectively. White zones in FIG.106 may be interpreted as thermal damage, but IRE resistance decreased from a first to last pulse. Typically, when thermal damage occurs, the resistance decreases then rapidly increases as the tissue is desiccated. The same trend is observed in the Signature (sinusoid) magnitudes, i.e. a decrease in the resistance. After the decrease in the resistance, a subsequent increase is not observed, indicating little or no thermal damage. Observance of lesions after 4 hours in live tissue provides additional insight into the white zone.

[0421] FIGS.107-112 illustrate various plots and images. For example, FIG.107 depicts a phase of the clamped tissue before and after the IRE application for Group_set 1_1 and 1_2. FIG. 108 depicts a phase of the clamped tissue before and after the IRE application for Group_set 2_1 and 2_2. FIG.109 depicts a phase of the clamped tissue before and after the IRE application for Group_set 3_1 and 3_2. FIG.110 depicts a phase of the clamped tissue before and after the IRE application for Group_set 4_1 and 4_2. FIG.111 depicts treatment zones for 2,000 and 3,000 pulses for BiClamp7 device. FIG.112 depicts treatment zones for 800 and 1,700 pulses for BiClamp8 device.

[0422] This study records BiClamp applications on live porcine jejunum. There is a clear correlation of change in a slope of phase of the signature and a number of pulses applied. As is concluded with the articulating tip applicator 202, this is the best indicator of an extent of treatment with IRE. What remains is to correlate a depth of treatment with phase changes in a lab in which animal survives for at least 24 hours for full resolution of the lesions.

[0423] The signature is not limited to this definition, rather it also may be defined as a combination of the magnitude and phase of the sinusoidal signal before and after IRE treatment.

[0424] It is to be understood that the invention is not limited in its application to the details of construction and the arrangement of the components set forth herein. The invention is capable of other embodiments and of being practiced or being carried out in various ways. Variations and modifications of the foregoing are within the scope of the present invention. It should be understood that the invention disclosed and defined herein extends to all alternative combinations of two or more of the individual features mentioned or evident from the text and / or drawings. All of these different combinations constitute various alternative aspects of the present invention. The embodiments described herein explain the best modes known for practicing the invention and will enable others skilled in the art to utilize the invention.

[0425] While the preferred embodiments of the invention have been shown and described, it will be apparent to those skilled in the art that changes and modifications may be made therein without departing from the spirit of the invention, the scope of which is defined by this description.

Claims

Arnold & Porter Ref. P35370WO00 CLAIMS:

1. A device for non-thermal treatment of one or more lesions in a subject in need thereof comprising: a generator 110 comprising a first output connector and a second output connector; an applicator 200, wherein applicator 200 is a articulating tip applicator 202 comprising a first electrode 210 and a second electrode 220; a dispersive pad 300; and a switch 410 in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator.

2. The device of claim 1, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad.

3. The device of claim 2, wherein the switch 410 in the first position is configured to provide an electrical connection between the generator 110 through the first output connector with the first electrode 210 and the second electrode 220 of the applicator and provide an electrical connection between the generator through the second output connector with the dispersive pad 300.

4. The device of claim 3, wherein the first electrode 210 and the second electrode 220 are in electrical connection with each other.

5. The device of claim 3, wherein the dispersive pad 300 is configured to be in contact with the patient when the electric field is applied.

6. The device of claim 5, wherein the applicator 200 is configured to operate as a blunt tip for the treatment of the one or more lesions.

7. The device of claim 5, wherein the dispersive pad 300 is a large surface electrode.

8. The device of claim 1, wherein the second mode is a bipolar mode that generates an electric field between the first electrode 210 and the second electrode 220 of the applicator.Arnold & Porter Ref. P35370WO00 9. The device of claim 1, wherein the switch in the second position is configured to provide an electronic connection between the generator 110 through the first output connector with the first electrode 210 and provide an electronic connection between the generator 110 through the second output connector with the second electrode 220.

10. The device of claim 8, wherein there is no electrical connection between the generator 110 and the dispersive pad 300.

11. The device of claim 1, wherein the electric field of the first mode through the applicator 300 is capable of forming a treatment zone comprising the one or more lesions.

12. The device of claim 1, wherein the electric field of the second mode through the applicator is capable of forming a treatment zone comprising the one or more lesions.

13. The device of claim 1, wherein the first electrode 210 comprises a first stainless steel rod 212 surrounded by a first insulating layer 212i, a distal end 213, and a proximal end 214, wherein the second electrode 220 comprises a second stainless steel tube 222 surrounded by a second insulating layer 222i, a distal end 223, and a proximal end 224, wherein the first electrode 210 and the second electrode 220 are concentric, and wherein the proximal end 214 of the first electrode and the distal end 223 of the second electrode are separated by a distance in length 230w.

14. The device of claim 13, further comprising an insulating layer 230 between the first electrode 210 and the second electrode 220.

15. The device of claim 14, wherein the insulating layer 212 extends beyond the distal end of the first electrode by between 0 and 2 mm.

16. The device of claim 13, wherein the first electrode comprises a first outer diameter 216 of between 2 and 4 millimeters (mm).

17. The device of claim 13, wherein the first outer diameter 216 is approximately 3 mm.

18. Device of claim 13, wherein the second electrode 220 comprises a second outer diameter 226 of between 6 and 10 mm.

19. The device of claim 13, wherein the first electrode 210 comprises a wall thickness between 0 mm and 0.8 mm.Arnold & Porter Ref. P35370WO00 20. The device of claim 13, wherein the distance in length between the proximal end 214 of the first electrode and the distal end 223 of the second electrode is between 0 mm and 6 mm.

21. A device for an application of a non-thermal treatment dose comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a first electrode and a second electrode; a first switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply a therapeutic electric field of a first mode through the applicator and a second position to apply a therapeutic electric field of a second mode through the applicator; a dispersive pad in an electrical connection with the second output connector; a sensor configured to receive a first signal before the application of a non-thermal treatment dose and a second signal after the application of the non-thermal treatment dose; a processor electronically coupled to the switch and the sensor, a non-transitory computer readable medium electronically coupled to the processor, wherein the non-transitory computer readable medium comprises instructions stored thereon that, when executed on the processor, perform the steps of receiving the first signal from the sensor and the second signal from the sensor; and determining a difference between the first signal and the second signal, comparing the difference against a predetermined threshold; and instructing the application to end if the difference exceeds the predetermined threshold.

22. The device of claim 21, wherein the first switch is in a first electrical connection with the first output connector and in a second electrical connection with the first electrode 210.

23. The device of claim 22, wherein the instructing the application to end comprises a third signal to a second switch 420 upstream of the first switch and in a first electrical connection with the first output connector and in a second electrical connection with the first electrode, wherein the second switch is configured to disrupt the first electrical connection to the first output connector and the second electrical connection with the first electrode.

24. A device for an application of a non-thermal treatment dose comprising: a generator 110 comprising a first output connector and a second output connector;Arnold & Porter Ref. P35370WO00 an applicator 200 comprising a first electrode 210 in a first electrical connection with the first output and a second electrode 220 in a second electrical connection with the second output; a switch in a first electrical connection with the first output connector and in a second electrical connection with the first electrode, wherein the switch in a first position is configured to apply a therapeutic electric field through the first electrode, and wherein the switch in a second position is configured to disrupt the first electrical connection to the first output connector and the second electrical connection with the first electrode; a sensor configured to receive a first signal before the application of a non-thermal treatment dose and a second signal after the application of the non-thermal treatment dose; a processor electronically coupled to the switch and the sensor; a non-transitory computer readable medium electronically coupled to the processor, wherein the non-transitory computer readable medium comprises instructions stored thereon that, when executed on the processor, perform the steps of receiving the first signal from the sensor and the second signal from the sensor; and determining a difference between the first signal and the second signal, comparing the difference against a predetermined threshold; and 25. instructing the switch to configure to the second position. A device for non-thermal treatment comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a first electrode and a second electrode; a dispersive pad; and a first switch 410 in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator; a second switch 420 upstream of the first switch and in a first electrical connection with the first output connector and in a second electrical connection with the first electrode, wherein the second switch is configured to disrupt the first electrical connection to the first output connector and the second electrical connection with the first electrode; a sensor configured to recognize a signal; a processor electronically coupled to the switch and configured to receive a second signal from the sensor; andArnold & Porter Ref. P35370WO00 a non-transitory computer readable medium electronically coupled to the processor, wherein the non-transitory computer readable medium comprises instructions stored thereon activated, after the receipt of the signal, to perform the steps of disconnecting the electrical connection between the first electrical connection and the first output.

26. The device of claim 21, 24, or 25, wherein the signal comprises a measurement for electrical magnitude and phase.

27. A method of non-thermal treatment of a lesion in a subject in need thereof comprising: identifying the location, size and volume of the lesion in the subject; determining an electric field useful in the treatment of the identified lesion; and applying the electric field to the lesion for an appropriate time and appropriate voltage using a device comprising: a generator comprising a first output connector and a second output connector; an applicator comprising a first electrode and a second electrode; a dispersive pad; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator.

28. The method of claim 27, wherein the first mode is a monopolar mode that generates aa electric field between the applicator and the dispersive pad.

29. The method of claim 28, wherein the dispersive pad is in contact with the subject in need thereof.

30. A method of non-thermal treatment of a lesion in a subject in need thereof comprising: determining an electric field useful in the treatment of the lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the lesion with a device comprising: a generator comprising a first output and a second output; an applicator comprising a first electrode and a second electrode; a dispersive pad; andArnold & Porter Ref. P35370WO00 a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply the electric field of the first mode through the applicator and a second position to apply the electric field of the second mode through the applicator, wherein the electric field is within a treatment area in the subject in need thereof.

31. The method of claim 30, wherein the lesion is an endometrial lesion, a malignant skin lesion, a non-malignant skin lesion, a bladder lesion, a ductal breast cancer lesion, a lesion of the gastrointestinal system, or a combination thereof.

32. The method of claim 30, wherein the electric field does not cause damage outside of the treatment area.

33. The method of claim 30, wherein the applying the determined electric field comprises applying the determined electric field in cycles separated by one or more rest periods to reduce thermal damage.

34. The method of claim 30, wherein the electric field comprises 500 to 2000 volts (V).

35. The method of claim 30, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad.

36. The method of claim 35, wherein the dispersive pad is configured to be in contact with the patient when the electric field is applied.

37. The method of claim 30, wherein the second mode is a bipolar mode that generates an electric field between the first electrode and the second electrode of the applicator.

38. A method of causing controlled cell necrosis in an endometrium of a subject in need thereof comprising: determining an electric field useful in the treatment of an endometrial lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the endometrial lesion with a device comprising: a generator comprising a first output and a second output, an applicator comprising a first electrode and a second electrode; a dispersive pad; andArnold & Porter Ref. P35370WO00 a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply the electric field of the first mode through the applicator and a second position to apply the electric field of the second mode through the applicator, wherein the electric field of the first mode or the electric field of the second mode is within a treatment area of the subject in need thereof and causes cell necrosis within the treatment area.

39. The method of claim 38, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad.

40. The method of claim 39, wherein the dispersive pad is configured to be in contact with the patient when the electric field is applied.

41. The method of claim 38, wherein the second mode is a bipolar mode that generates an electric field between the first electrode and the second electrode of the applicator.

42. The device of claim 1, wherein the applicator is an articulating tip catheter comprising a shaft comprising: a distal end comprising a first electrode 210 comprises a first stainless steel rod 212, a distal end 213, and a proximal end 214; a second electrode 220 comprises a second stainless steel tube 222, a distal end 223, and a proximal end 224, and an insulating layer; a flexible section; and a proximal end comprising mechanical levers connected to the flexible section with cables, wherein the flexible section is situated between the proximal end and the distal end, and wherein the first electrode 210 and the second electrode 220 are concentric, and the proximal end 214 of the first electrode and the distal end 223 of the second electrode are separated by a distance in length 230w.

43. An articulating tip applicator comprising: a shaft comprising: a distal end comprising a first electrode 210 comprises a first stainless steel rod 212, a distal end 213, and a proximal end 214; a second electrode 220 comprises a second stainless steel tube 222, a distal end 223, and a proximal end 224, and an insulating layer;Arnold & Porter Ref. P35370WO00 a flexible section; and a proximal end comprising mechanical levers connected to the flexible section with cables, wherein the flexible section is situated between the proximal end and the distal end, and wherein the first electrode 210 and the second electrode 220 are concentric, and the proximal end 214 of the first electrode and the distal end 223 of the second electrode are separated by a distance in length 230w.

44. The articulating tip applicator of claim 43, wherein the proximal end of the shaft further comprises a motor.

45. A device for non-thermal treatment of one or more lesions in a subject in need thereof comprising: a generator 110 comprising a first output connector and a second output connector; an applicator 1200 comprising a first electrode 1210 and a second electrode 1220; a dispersive pad 300; and a switch 410 in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply an electric field of a first mode through the applicator and a second position to apply an electric field of a second mode through the applicator.

46. The device of claim 45, wherein the switch 410 is selected from the group consisting of a mechanical switch, an electronic switch, a latch switch, a momentary switch, a double throw switch, a single pole switch, or a changeover switch.

47. The device of claim 46, wherein the first mode is a monopolar mode that generates an electric field between the applicator and the dispersive pad.

48. The device of claim 46, wherein the switch 410 in the first position is configured to provide an electrical connection between the generator 110 through the first output connector with the first electrode 1210 and the second electrode 1220 of the applicator and provide an electrical connection between the generator through the second output connector with the dispersive pad 300.

49. The device of claim 46, wherein the dispersive pad 300 is configured to be in contact with the patient when the electric field is applied.Arnold & Porter Ref. P35370WO00 50. A device for non-thermal treatment of one or more lesions in a subject in need thereof comprising an applicator 1200 comprising: a proximal end 1201; a shaft 1203; applicator tip 1205 comprising one or more first electrode 1210 and one or more second electrode 1220; and distal end 1209.

51. The device of claim 50, further comprising two or more of the first electrode 1210 and two or more of the second electrode 1220.

52. The device of claim 51, wherein the two or more of the first electrode 1210 and two or more of the second electrode 1220 are separated by insulating layer 1290.

53. A method of non-thermal treatment of a lesion in a subject in need thereof comprising: determining an electric field useful in the treatment of the lesion to form a determined electric field; applying the determined electric field of a first mode or an electric field of a second mode to the lesion with a device comprising: a generator comprising a first output and a second output; an clamping probe comprising a first probe and a second probe; and a switch in electrical connection with the generator, wherein the switch is capable of changing between a first position to apply the electric field of the first mode through the applicator and a second position to apply the electric field of the second mode through the applicator, wherein the electric field is within a treatment area in the subject in need thereof.

54. The method of claim 53, wherein the device further comprises a dispersive pad.

55. The method of claim 53, wherein the clamping probe is clamping probe 201, 301, or 401.

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