Electrochemotherapy increases local control after incomplete excision of a recurring penile fibrosarcoma in a stallion.

An eleven-year-old stallion was referred for adjuvant treatment of an incompletely excised, recurring penile fibrosarcoma. The horse was bright, alert and responsive with a 15 x 12 cm ulcerated lesion on the ventral side of the penis. The lesion was the tumor bed of an incompletely excised fibrosarcoma. After complete staging procedures, the owner elected to treat the horse with electrochemotherapy (ECT) using cisplatin as chemotherapy agent. Two sessions of ECT were performed at two-week intervals using local cisplatin followed by trains of biphasic electric pulses applied using different electrodes until complete coverage of the area was achieved. The treatment was well tolerated, and the patient is still disease free after 12 months. ECT resulted in improved local control and should be considered among the available adjuvant treatments in equines carrying soft tissue tumors.

A case of advanced second-degree atrioventricular block in a ferret secondary to lymphoma.

A female ferret was referred as an emergency for severe respiratory distress symptoms. At presentation, the patient was listlessness, dyspnoeic, and hyper-responsive. The clinical examination evidenced dyspnea with cyanosis, altered cardiac rhythm, and hepatomegaly. Electrocardiography showed an advanced second-degree atrioventricular (AV) block. The liver aspirate was diagnostic for lymphoma. The patient did not respond to supportive therapy and rapidly died. Post-mortem exams confirmed the presence of lymphoma with hepatic involvement. Moreover, a pericardial lymphocytic infiltration and a widespread myocardial nodular localization of lymphoma were evidenced as well. This condition was probably the cause of the cardiac arrhythmia. To the best of our knowledge, ours is the first report of cardiac lymphoma causing heart block in ferrets.

Electrochemotherapy as First Line Cancer Treatment: Experiences from Veterinary Medicine in Developing Novel Protocols.

Tumor microenvironment is one of the major obstacles to the efficacy of chemotherapy in cancer patients. The abnormal blood flow within the tumor results in uneven drug distribution. Electrochemotherapy (ECT) is a tumor treatment that adopts the systemic or local delivery of anticancer drugs with the application of permeabilizing electric pulses having appropriate amplitude and waveforms. This allows the use of lipophobic drugs that frequently have a narrow therapeutic index maintaining at the same time a reduced patient morbidity and preserving appropriate anticancer efficacy. Its use in humans is addressed to the treatment of cutaneous neoplasms or the palliation of skin tumor metastases, and a standard operating procedure has been devised. On the other hand, in veterinary oncology this approach is gaining popularity, thus becoming a first line treatment for different cancer histotypes, in a variety of clinical conditions due to its high efficacy and low toxicity. This review summarizes the state of the art in veterinary oncology as a preclinical model and reports the new protocols in terms of drugs and therapy combination that have been developed.

Electroporation Enhances Bleomycin Efficacy in Cats with Periocular Carcinoma and Advanced Squamous Cell Carcinoma of the Head.

BACKGROUND: Advanced carcinoma of the head represents a substantial health problem in cats for local control and overall survival. OBJECTIVES: Evaluate the capability of electrochemotherapy (ECT) to improve bleomycin efficacy in cats with periocular carcinoma and advanced carcinoma of the head. ANIMALS: Twenty-one cats with periocular carcinoma (17 squamous cell carcinoma [SCC] and 4 anaplastic carcinoma) and 26 cats with advanced SCC of the head. METHODS: Nonrandomized prospective controlled study. Periocular carcinoma cohorts: 12 cats were treated with bleomycin (15 mg/m(2) i.v.) coupled with ECT under anesthesia; 9 cats were treated with bleomycin alone. Advanced head SCC cohorts: 14 cats were treated with bleomycin (15 mg/m(2) i.v.) coupled with ECT administered under sedation; 12 control cats were treated with bleomycin alone. ECT treatments (2-8) were performed every other week until complete remission (CR) or tumor progression occurred. RESULTS: Toxicities were minimal and mostly treated symptomatically. Overall response rate in the ECT treated animals was 89% (21 Complete Response [CR] and 2 Partial Response [PR]) whereas controls had response rate of 33% (4 CR and 3 PR). Median time to progression in ECT group was 30.5 months, whereas in controls it was 3.9 months (P < .0001). Median time to progression for ECT cohorts was 24.2 months for periocular cohort and 20.6 in advanced head SCC cohort, respectively. CONCLUSIONS: Electrochemotherapy is well tolerated for advanced SCC of the head in cats; its use may be considered among loco-regional strategies for cancer therapy in sensitive body regions such as periocular region.

Evaluation of Cisplatin as an electrochemotherapy agent for the treatment of incompletely excised mast cell tumors in dogs.

BACKGROUND: Electrochemotherapy (ECT) couples the administration of anticancer drugs with the delivery of electric pulses that increase the drug uptake through the cell membranes, resulting in an improved efficacy. HYPOTHESIS: To evaluate the tolerability and efficacy of cisplatin (CDDP) as an ECT agent to prevent recurrence of incompletely resected mast cell tumors (MCTs). ANIMALS: Thirty-seven dogs. METHODS: Prospective study recruiting dogs with incompletely excised MCTs as confirmed by surgeon and pathology reports. After debulking, the tumor bed and margins were infiltrated with CDDP, and then exposed to trains of biphasic electrical pulses under sedation. Five minutes after the injection of the chemotherapy agent, sequences of 8 biphasic pulses lasting 50 + 50 µs each, were delivered in bursts of 1,300 V/cm for sclerosed and of 800 V/cm for exposed lesions, with caliper or needle array electrodes, respectively. A second session was performed 1 or 2 weeks later based on clinical considerations. RESULTS: The treatment was well tolerated with minimal adverse effects. Twenty-nine dogs had no evidence of recurrence over the 6-year study period, 6 had tumor recurrence, 1 died of multiple cutaneous MCTs, and 1 died of unrelated causes. The estimated median time to recurrence was 1,200 days. Recurrence was not observed among the long-term (> 1 year) treated dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: ECT with CDDP appears effective in the treatment of incompletely resected MCT in dogs and could be a useful addition to the current options based on its low cost, limited toxicity, and ease of administration.

Progressive multifocal leukoencephalopathy following chemotherapy for lung cancer.

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system caused by the JC virus infection and with a frequent lethal outcome. PML usually occurs in immunocompromised subjects, such as HIV-positive individuals, as well as in other conditions characterized by depletion of cellular immunity, including hematological malignancies, autoimmune diseases, and immunomodulatory therapies. We describe the case of a 76-year-old man affected by advanced non-small cell lung cancer who developed PML after six cycles of carboplatin/gemcitabine therapy, during which a transitory leucopenia developed. The patient deceased a few months after the onset of the symptoms. Chemotherapy appears to be uncommon, but definite condition associated to PML.

Electrochemotherapy treatment for bilateral pleomorphic rhabdomyosarcoma in a cat.

A 13-year-old male neutered cat was presented for the sudden growth of two nodular lesions close to the upper eyelid of both eyes. Fine-needle aspiration cytology was suggestive of mesenchymal neoplasia. The cat had conservative surgical excision in order to preserve the eyelids' functionality; however, the histopathological report came with a diagnosis of incompletely excised bilateral pleomorphic rhabdomyosarcoma. Due to the local aggressiveness of this neoplasm, the cat was treated with two sessions of cisplatin-based electrochemotherapy, delivered 14 days apart. Systemic or local toxicities were not detected during the whole course of therapy. The cat is still in complete remission after 12 months. Electrochemotherapy is a safe and efficacious adjuvant therapy for aggressive sarcomas and warrants further investigations in order to standardise its protocols.

Zoledronic acid for the treatment of appendicular osteosarcoma in a dog.

A 10-year-old male intact Corso dog was referred for lameness and for a large neoplasm affecting the right foreleg. Physical examination of the patient revealed a 5 x 5 x 3 cm mass in the distal right foreleg. Histopathology was consistent with a diagnosis of appendicular osteosarcoma. The staging process found no evidence of metastasis. Because of the large size of the patient, the owners elected to treat their dog with antiresorptive therapy. The patient was treated with an infusion of zoledronic acid every 28 days. The tumour remained stable for 16 months and the lameness of the dog greatly improved. At that time, the patient returned for evaluation of a large rapidly growing prescapular mass. Biopsy confirmed lymph node metastasis and the dog was euthanased. Zoledronic acid showed remarkable palliation in our patient and possibly anti-tumour action and warrants further investigation.

Expression patterns of two serine protease HtrA1 forms in human placentas complicated by preeclampsia with and without intrauterine growth restriction.

Preeclampsia (PE) and intrauterine growth restriction (IUGR) are pregnancy-specific disorders that have in common abnormal placental implantation, a marked proliferation of villous cytotrophoblastic cells and focal necrosis of the syncytiotrophoblast. Several studies show an ischemic placenta with a high-resistance vasculature, which cannot deliver an adequate blood supply to the feto-placental unit. The cause of PE is a matter of debate, but recently studies in mice suggest that the primary feto-placental lesions are sufficient to initiate the disease. HtrA1, a member of the family of HtrA proteins, is a secreted multidomain protein with serine protease activity. It is expressed in first and third trimester of gestation. In specimens from the first trimester of gestation, immunostaining for HtrA1 is generally found in both layers of villous trophoblast, syncytiotrophoblast and cytotrophoblast. Cytoplasm of extravillous trophoblast and extracellular matrix of cell islands and cell columns are labeled for HtrA1. Specimens from third trimester of gestation show a more intense positivity for HtrA1 in the syncytiotrophoblast than in cytotrophoblast. The extravillous trophoblast and the decidual cells, is positive for HtrA1. The purpose of this study is to investigate the expression pattern of HtrA1 in placentas from PE without IUGR (maternal PE) and with IUGR (fetal PE) by quantitative western blotting and immunohistochemistry. By quantitative western blotting analysis we observed a significant upregulation of approximately 30 kDa HtrA1 form in PE. Differently, we detected a significant total HtrA1 down-regulation in PE-IUGR. Moreover, immunostaining for HtrA1 was positive in the villous trophoblast, in the syncytial knots and irregularly in the fetal vessel walls in PE placentas while immunostaining for HtrA1was present particularly in the syncytial knots in PE-IUGR placentas. In conclusion, we suggest that the approximately 30 kDa HtrA1 form can be correlated to maternal PE while that the significant down-regulation of total HtrA1 can be correlated to placental PE. These HtrA1 alterations could be considered as possible markers to discriminate placental PE from maternal PE.

The Helicobacter pylori's protein VacA has direct effects on the regulation of cell cycle and apoptosis in gastric epithelial cells.

In this study, we have evaluated the effects on cell cycle regulation of VacA alone and in combination with other two Helicobacter pylori proteins, cytotoxin-associated protein (CagA) and HspB, using the human gastric epithelial cells (AGS). Our results indicate that VacA alone was able to inhibit the G1 to S progression of the cell cycle. The VacA capacity of inhibiting cell progression from G1 to S phase was also observed when cells were co-transfected with CagA or HspB. Moreover, VacA over-expression caused apoptosis in AGS cells through activation of caspase 8 and even more of caspase 9, thus indicating an involvement of both the receptor-mediated and the mitochondrial pathways of apoptosis. Indeed, the two pathways probably can co-operate to execute cell death with a prevalence of the mitochondrial pathways. Our data taken together provide additional information to further enhance our understanding of the molecular mechanism by which H. pylori proteins alter the growth status of human gastric epithelial cells.

Electrochemotherapy for localized lymphoma: a preliminary study in companion animals.

Non Hogdkin's lymphoma is the commonest malignant neoplasm in humans and in pets. Treatments include systemic chemotherapy eventually combined with radiation therapy. Radiation therapy is also used as single agent for the treatment of localized lymphoma (LSA). Albeit efficacious, this modality is potentially associated with side effects. Purpose of this study was to preliminarily evaluate the feasibility and efficacy of electrochemotherapy (ECT) in companion animals with localized lymphoma. Six patients entered the study and received two sessions of ECT under sedation. The pets had local injection of bleomycin at the concentration of 1.5 mg/mg and five minutes after the chemotherapy, trains of 8 biphasic electric pulses lasting 50 + 50 micros each, with 1 ms interpulse intervals, were delivered by means of modified caliper electrodes or for difficult districts, through paired needle electrode. All the patients achieved complete responses (lasting 1 week through 3 years), one cat with nasal LSA had local recurrence and two others experienced spinal and intestinal relapse. Side effects were not noted with the exception of focal alopecia in a cat with retrobulbar LSA. Electrochemotherapy appears as a safe and efficacious modality for the treatment of localized lymphoma and warrants further investigations.

Inhibition of the SH3 domain-mediated binding of Src to the androgen receptor and its effect on tumor growth.

In human mammary and prostate cancer cells, steroid hormones or epidermal growth factor (EGF) trigger association of the androgen receptor (AR)-estradiol receptor (ER) (alpha or beta) complex with Src. This interaction activates Src and affects the G1 to S cell cycle progression. In this report, we identify the sequence responsible for the AR/Src interaction and describe a 10 amino-acid peptide that inhibits this interaction. Treatment of the human prostate or mammary cancer cells (LNCaP or MCF-7, respectively) with nanomolar concentrations of this peptide inhibits the androgen- or estradiol-induced association between the AR or the ER and Src the Src/Erk pathway activation, cyclin D1 expression and DNA synthesis, without interfering in the receptor-dependent transcriptional activity. Similarly, the peptide prevents the S phase entry of LNCaP and MCF-7 cells treated with EGF as well as mouse embryo fibroblasts stimulated with androgen or EGF. Interestingly, the peptide does not inhibit the S phase entry and cytoskeletal changes induced by EGF or serum treatment of AR-negative prostate cancer cell lines. The peptide is the first example of a specific inhibitor of steroid receptor-dependent signal transducing activity. The importance of these results is highlighted by the finding that the peptide strongly inhibits the growth of LNCaP xenografts established in nude mice.

Cell-cycle molecules in mesothelioma: an overview.

Cell cycle progression is mediated by a group of proteins named cyclins that activate a highly conserved family of protein kinases, the cyclin-dependent kinases (CDKs). CDKs are also regulated by related proteins called cdk inhibitors, grouped into two families: the INK4 inhibitors (p16, p15, p19 and p18) and the Cip/Kip inhibitors (p21, p27). Moreover, several tumour suppressor genes (such as Retinoblastoma gene and p53 gene) are implicated in the regulation of the molecular mechanism of cell division. Several studies report the importance of cell cycle regulator proteins in the pathogenesis and the prognosis of mesothelioma. This article will review the most recent data from the literature about the expression and the diagnostic and prognostic significance of cell cycle molecules in mesothelioma.

Biphasic pulses enhance bleomycin efficacy in a spontaneous canine perianal tumors model.

Perianal tumors (adenoma and carcinoma of the hepatoid glands) are frequently reported in veterinary literature. They are locally aggressive tumors with a low tendency to metastatic spread. An hormonal ethiology has been identified for the development of perianal adenomas in male dogs, while the carcinomas are free from hormonal influence. Standard treatments include surgery, cryotherapy or, in selected cases, radiation therapy. In this article we describe the outcome of a small cohort of canine patients with perianal tumors treated with bleomycin selectively driven by trains of biphasic pulses (electrochemotherapy). Twelve canine patients, eight with adenoma and four with carcinoma of the perianal glands, entered the study and received two sessions of ECT under sedation. The pets had local injection ofbleomycin at the concentration of 1.5 mg/mg and five minutes after the chemotherapy, trains of 8 biphasic electric pulses lasting 50 + 50 micros each, with 1 ms interpulse intervals, were delivered by means of modified caliper and needle array electrodes or, for difficult districts, through paired needle electrode. The overall response rate was 91% with a 83% of complete response (10/12); one dog had a PR that lasted 12 months and another had progressive disease. Electrochemotherapy appears as a safe and efficacious modality for the treatment of perianal tumors and warrants further investigations.

Lomustine for the treatment of gastrointestinal mast cell tumour in a dog.

An eight-year-old, male boxer dog was referred for the treatment of a large (5.5 x 5 cm), unresectable visceral mast cell tumour. The dog had a surgical resection performed one month before referral, and it had widespread metastases to the abdominal lymph nodes. The patient was treated with lomustine and prednisone and showed a rapid improvement and increased level of activity, weight gain and consistent tumour reduction. The patient remained in partial remission (defined as a greater than 50 per cent reduction in tumour volume) for seven months. Toxicity was acceptable and was limited to moderate anaemia and two episodes of neutropenia. At the completion of the seven months of therapy, the dog experienced a chemotherapy-induced sepsis, and the owners elected for euthanasia due to financial concerns. At that time, the tumour was still in partial remission. This case report suggests that a combination of lomustine and prednisone is an effective protocol for the palliation of aggressive visceral mast cell tumours.