PMC9363929_01
The photograph in (a) depicts the appearance of a large mass that involved the right orbit and at the time of presentation had erupted through the skin as an open wound over the bridge of the nose and face. The photomicrograph in (b) depicts the prominent immunohistofluorescent labeling of VGSCs (green) observed in a tissue sample taken from the tumor in (a) before treatment with TOL. For comparison, the photomicrograph in (c) depicts a similarly processed tissue sample showing VGSC expression in a murine homograft that was assessed in a pre-clinical study to determine the efficacy of TOL in treating triple-negative breast cancer. The calibration bar for photomicrographs in B = 25 mum and nuclei (blue) are labeled with DRAQ5.
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fig2
The patient presented with a 10-month history of intermittent dysphagia, rhinitis and epistaxis, epiphora, nasal swelling, and weight loss and displayed a prominent mass that had erupted through the skin as an open wound over the bridge of the nose and face and involved the right orbit (Figure 2(a)).
The CT images depict soft tissue (b, d) and bone window (a, c) images of the rostral (a, b) and caudal (c, d) portions of the tumor shown in Figure 1 prior to treatment with TOL. Note that the tumor involves and extends beyond the margins of the right orbit, invades the paranasal sinuses bilaterally, and perforates the hard palate extending into the oral pharynx.
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fig3
A CT scan revealed a large tumor that occupied the right nasal cavity, had invaded the orbit and nasal turbinate bones, and extended inferiorly to involve the hard palate (Figure 3).
PMC9363929_02
The CT images depict soft tissue and bone window images of closely comparable rostral (a-f) and caudal (g-l) portions of the tumor shown before (a, b, g, h) and after (c, d, i, j, e, f, k l) each of 2 rounds of treatment with TOL. Despite the improvement in the patient's level of social interaction and appetite, there was no evidence to support a reduction in the tumor mass.
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fig4
CT imaging obtained 1 week after treatment indicated that the size of the tumor had not changed (Figure 4).
The photograph in (a) depicts the appearance of the mass presented Figure 1 after 2 rounds of treatment with TOL. Note the reduction in localized edema revealing more of the globe of the eye. The photomicrograph in (b), compared to the photomicrograph in Figure 1(b), depicts the relative absence of cells that highly express VGSCs (green) following treatment with TOL. The calibration bar = 25 mum.
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fig5
By contrast, the area of tumor visible on the surface had decreased sufficiently to allow more of the orbit to be discernible (Figure 5(a)). A posttreatment biopsy revealed very few cells that highly expressed VGSCs (Figure 5(b)).
PMC9363929_03
PMC9363929_04
The CT images depict soft tissue (b, d) and bone window (a, c) images of the rostral (a, b) and caudal (c, d) portions of a large facial carcinoma that involved the right orbital wall and nasal turbinate bones and had extended into the paranasal sinuses.
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fig6
The patient was referred for treatment of a large facial carcinoma that involved the left orbital wall and nasal turbinate bones and had extended into the paranasal sinuses (Figures 6 and 7(a)).
The photograph in (a) depicts the appearance of a large mass that involved the right orbit and at the time of presentation had erupted through the skin over the bridge of the nose and face. The low-power (b) and high-power (c) photomicrographs depict the prominent immunohistofluorescent labeling of VGSCs (green) revealed in a tissue sample taken from the tumor in (a) before treatment with TOL. The calibration bar for photomicrographs in (b) and (c) is 50 and 25 mum, respectively.
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fig7
The patient was referred for treatment of a large facial carcinoma that involved the left orbital wall and nasal turbinate bones and had extended into the paranasal sinuses (Figures 6 and 7(a)). VGSC expression revealed in a tissue biopsy indicated the likelihood of a favorable response to treatment with TOL (Figures 7(b) and 7(c)).
PMC9363929_05
The photograph in (a) depicts the toroidal device used to deliver the uniform PEFs with to open VGSCs; maximum field amplitude of 6 V/m. The photograph in (b) depicts the coaxial ring device used to deliver PEFs with a field amplitude of 18 V/m. (c) Dogs in the carriers that allowed room for ad libitum movement to adjust position for comfort during the period of stimulation while being treated with TOL.
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fig1
Two weeks later, a fifth and final round in the initial series of treatments with TOL was administered using a newly designed coaxial ring device (Figures 1(b) and 1(c)) that made it possible to deliver a uniform PEF amplitude of 18 V/m.
The photograph in (a) depicts the appearance of the mass presented in Figure 7 after debulking, cauterization and 4 rounds of treatment with TOL. The photomicrograph in (b), compared to the photomicrograph in Figure 7(b) and Figure 7(c), depicts the significant reduction in the number of cells that highly express VGSCs (green) following treatment with TOL. The calibration bar in (c) is 25 mum.
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fig8
The size of the rostral portion of the tumor had decreased and was no longer visible on gross examination (Figure 8(a)), but the caudal portion of the tumor continued to grow and destroy more of the facial bones.
PMC9363929_06
The photograph in (a) depicts the appearance of the mass presented in Figure 7 after debulking, cauterization and 4 rounds of treatment with TOL. The photomicrograph in (b), compared to the photomicrograph in Figure 7(b) and Figure 7(c), depicts the significant reduction in the number of cells that highly express VGSCs (green) following treatment with TOL. The calibration bar in (c) is 25 mum.
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fig8
The size of the rostral portion of the tumor had decreased and was no longer visible on gross examination (Figure 8(a)), but the caudal portion of the tumor continued to grow and destroy more of the facial bones.
The soft tissue (b, d) and bone window (a, c) CT images obtained approximately 6 months after the images presented in Figure 6 approximate the levels shown in Figure 6. Despite palliative debulking procedures and 6 months of chemotherapy, no appreciable changes were noted in the deep-seated portions of the tumor.
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fig9
Despite palliative debulking procedures, a CT scan obtained after the 6 months of chemotherapy revealed that there had been no appreciable change in the size of the deep-seated portions of the tumor (Figure 9).
PMC9363929_07
The chest X-rays in (a) and (b) depict the AP and RL appearance of 2 masses in the right lung prior to treatment with TOL. The larger mass (black arrows) is located centrally in the middle lobe in close proximity to the hilus of the lung and smaller mass (white arrows) is located more caudal in the cranial lobe. (c, d) The appearance of the same masses 7 months later after receiving 7 rounds of treatment with TOL. Taking account of the positioning in the X-ray machine, the size of the tumors varied but appeared to have increased gradually by approximately 28% over the 7 1/2 month course of treatment. The calibration bar in (d) is 2.5 mum.
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fig10
The patient was referred for evaluation and treatment of 2 masses that had been discovered incidentally in the right lung (Figure 10).
PMC9363929_08
The photomicrograph in (a) depicts the immunohistofluorescent labeling of VGSCs (green) observed in a tissue sample taken from the larger tumor mass before treatment with TOL. The immunohistofluorescent evaluation of a tissue sample obtained after treatment with TOL in B reveals the significant reduction in the number of cells that highly express VGSCs. The calibration bar in (b) is 25 mum.
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fig11
TOL (Figure 11(a)). Three weeks after pretreatment X-rays were obtained, treatment with TOL was initiated and administered without signs of adverse cognitive or behavioral effects.
PMC9363929_09
The photomicrograph in (a) is a low-power (X1) image of a representative portion of the lung tumor that had received 8 rounds of treatment with TOL alone over 7 1/2 months. The tissue was taken at the time of necropsy and stained with hematoxylin and eosin. Note the large areas of necrosis that comprise over 90% of the tumor mass. The photomicrograph in (b) is a X40 enlargement of the tissue field defined by the box in (a) that illustrates in greater detail the few blood vessels and neoplastic cells that remained.
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fig12
Gross and histopathologic analysis determined that necrosis, located both centrally and in portions of the tumor closer to its outer margins, comprised greater than 90% of the soft, white tumor (Figure 12(a)). Only a few blood vessels with adjacent neoplastic cells remained (Figure 12(b)). An additional sample was collected for immunohistochemical evaluation that revealed that most of the cells that highly expressed VGSCs were no longer present (Figure 12(b)), consistent with the interpretation that TOL had and was continuing to destroy the cells that had the highest levels of VGSC expression.
PMC9363929_10
The photographs in (a) depicts the appearance of a large tumor located in the left oral pharynx (large arrow) and a second, smaller mass on the right (small arrow) that was discovered on initial physical examination (a). The photograph in (b) depicts the tumors midway through a third treatment with TOL. Note that the smaller tumor is no longer apparent (small arrow) and the larger tumor is significantly reduced. The calibration bar in (a) is 3 mm. The photomicrograph in (c) depicts the immunohistofluorescent labeling of VGSCs observed in a tissue sample taken from the tumor in (a) before treatment with TOL. Note that TOL was the only treatment provided in the management of the tumors. The calibration bar in (c) is 25 mum.
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fig13
Examination revealed the presence of prominent oral mass occupying the left posterior lateral oral pharynx involving the soft palate, the upper and lower molar teeth, and extending anteriorly along the lateral margin of the tongue (Figure 13(a)). Upon consideration of the results achieved in the previous cases, a biopsy was obtained that revealed VGSC expression sufficiently high to anticipate a favorable response to treatment with TOL (Figure 13(c)) and a trial treatment was arranged. TOL revealed that the main tumor mass was much smaller and appeared "much less angry" than when first observed (Figure 13(b)) and the smaller tumor mass was no longer apparent. The cat has completed 6 rounds of treatment and continues to do well 5 months after the tumor was discovered.