Preclinical and Clinical Characterization of Fibroblast-derived Neuregulin-1 on Trastuzumab and Pertuzumab Activity in HER2-positive Breast Cancer.

PURPOSE: To characterize expression of neuregulin-1 (NRG1), an HER3 ligand, in HER2-positive breast cancer and its relation with the efficacy of trastuzumab with or without pertuzumab. EXPERIMENTAL DESIGN: Characterization of NRG1 expression in tumor cell lines, in tumor specimens, and in cancer-associated fibroblasts (CAFs). Patient-derived CAFs were used to investigate NRG1 impact on the activity of trastuzumab with or without pertuzumab in HER2-positive breast cancer cells. The relationship between NRG1 expression and pathologic response to anti-HER2-based neoadjuvant therapy was assessed in a retrospective patient cohort and in the NeoSphere trial. RESULTS: NRG1 was expressed in HER2-positive breast cancer-derived fibroblasts at significantly higher levels than in cancer cells. NRG1 and the conditioned media (CM) from CAFs phosphorylated HER3 and AKT in cancer cells and mediated trastuzumab resistance. Stable genetic depletion of NRG1 from CAFs overcame trastuzumab resistance. Pertuzumab effectively suppressed trastuzumab resistance mediated by either NRG1 or CAF's CM. NRG1 engaged an epithelial-to-mesenchymal transition that was prevented by trastuzumab and pertuzumab. In clinical samples, stromal and/or tumor cell expression of NRG1 determined by immunohistochemistry was uncommon (13.2%) yet significantly linked with residual disease following trastuzumab-based neoadjuvant therapy. In the NeoSphere trial, the magnitude of the difference of pathologic complete response rates favoring the pertuzumab arm was higher in the NRG1-high group. CONCLUSIONS: CAF-derived NRG1 mediates trastuzumab resistance through HER3/AKT, which might be reverted by pertuzumab. In patients with HER2-positive breast cancer, high expression of NRG1 was associated to poor response to trastuzumab, but not in combination with pertuzumab.

Application of ozone therapy in the vestibulocochlear syndrome.

The aim of this study was to evaluate the efficacy of ozone therapy in the treatment of 50 patients with peripheral vestibulocochlear syndrome. Ozone was injected in the cervical region C2-C3, for 20 sessions. Evaluation criteria was based in the evolution of nystagmus, tinnitus, hearing loss and vertigo. Also, oxidative stress parameters were measured. Results demonstrated that patient improvements, according to vertigo, hearing loss, tinnitus and nystagmus, were of 90, 80, 65 and 100%, respectively. These patients were initially under condition of systemic oxidative stress, however, at the end of the study a redox balance was achieved. No side effects were observed.

[Rectal ozone therapy for patients with pulmonary emphysema]. / La ozonoterapia en pacientes con enfisema pulmonar.

BACKGROUND: Ozone therapy may stimulate antioxidant systems and protect against free radicals. It has not been used formerly in patients with pulmonary emphysema. AIM: To assess the effects of rectal ozone therapy in patients with pulmonary emphysema. MATERIAL AND METHODS: Sixty four patients with pulmonary emphysema, aged between 40 and 69 years, were randomly assigned to receive rectal ozone in 20 daily sessions, rectal medicinal oxygen or no treatment. Treatments were repeated three months later in the first two groups. At baseline and at the end of the study, spirometry and a clinical assessment were performed. RESULTS: fifty patients completed the protocol, 20 receiving ozone therapy, 20 receiving rectal oxygen and 10 not receiving any therapy. At baseline, patients on ozone therapy had significantly lower values of forced expiratory volume in the first second (fEV1) and fEV1/forced vital capacity. At the end of the treatment period, these parameters were similar in the three treatment groups, therefore they only improved significantly in the group on ozone therapy. No differences were observed in other spirometric parameters. CONCLUSIONS: Rectal ozone therapy may be useful in patients with pulmonary emphysema.

Therapeutic efficacy of topical OLEOZON® in patients suffering from onychomycosis.

The results of the use of ozonised sunflower oil (OLEOZON(®)) in the treatment of onychomycosis, based on its known antimycotic action and good skin tolerance, by means of a controlled randomised phase III assay are presented. A total of 400 outpatients were randomly divided into two groups: experimental, treated with topical OLEOZON(®), two times per day and control, treated also two times per day, with ketoconazole cream 2%, for 3 months. A patient was considered cured when the sick nails regained the normal colour, growth and thickness, with a negative mycological study. In the experimental group, a regression of signs was achieved from the first month of treatment, while in the control group, it was obtained after the third month of treatment. All patients treated with OLEOZON(®) had improvement in their condition (9.5%) or were cured (90.5%). However, in the control group, only 13.5% of patients were cured, 27.5% improved and 59% remained the same, with significant differences between both the groups. After 1 year of follow-up, experimental and control groups presented 2.8% and 44.4% of relapses, respectively. Topical OLEOZON(®) demonstrated effectiveness in the treatment of onychomycosis, superior to that of ketoconazole. No side effects were observed.

Ozone oxidative post-conditioning in acute renal failure.

OBJECTIVES: The ischaemia-reperfusion process is largely mediated by reactive oxygen species. Taking into account that a transient and controlled administration of ozone is able to upregulate cellular antioxidant enzymes, a morphological, biochemical and functional renal study was performed in rats undergoing warm renal ischaemia. METHODS: Rats were divided into four groups. All except the negative controls underwent 60 min' bilateral renal ischaemia followed by 10 days' reperfusion. The positive control group received no further treatment. The ozone group received an ozone/oxygen mixture (ozone dose 0.5 mg/kg) immediately after the ischaemia and daily for the 10 days' reperfusion; the oxygen group were given the same concentration of oxygen alone (13 mg/kg). Biochemical parameters fructosamine, phospholipase A2, catalase, superoxide dismutase and thiobarbituric acid reactive substances were measured, as well as renal plasma flow and glomerular filtration rate. KEY FINDINGS: Renal plasma flow and glomerular filtration rate decreased significantly in the positive controls and the oxygen group whereas values in the ozone group were similar to those in the negative control group. With respect to the biochemical parameters, ozone maintained a homeostasis redox, with significant increases in catalase and superoxide dismutase activities and similar values for phospholipase A2 and fructosamine compared with the negative control group. Fewer morphological alterations were seen in kidneys from the ozone group. No advantages were obtained in the positive control and oxygen groups. CONCLUSIONS: The protective effect of ozone may be explained by upregulation of the antioxidant defence system and beneficial effects on blood circulation and in oxygen metabolism. Ozone treatment may represent a therapeutic approach for minimising renal damage after transplantation.

Ozone therapy: clinical and basic evidence of its therapeutic potential.

Ozone has recently been subjected to criticism and emphasis in relation to clinical efficacy and toxicity, respectively. Without a doubt, ozone, in common with oxygen itself, is one of the most potent oxidants. Ozone is considered one of the major pollutants in urban areas. Nevertheless, increasingly widespread use lately has highlighted the potential benefits as a therapeutic agent when used according to well-defined and safe protocols. Basic studies conducted following rigorous scientific and ethical criteria have been proposed for scientific discussion. This paper concerns original data on an in vivo model of Parkinson's disease and published data on the effect of low ozone doses with any risk of toxicity excluded with the concentrations commonly used in medical applications.

Ozone oxidative preconditioning is mediated by A1 adenosine receptors in a rat model of liver ischemia/ reperfusion.

The liver is damaged by sustained ischemia in liver transplantation, and the reperfusion after ischemia results in further functional impairment. Ozone oxidative preconditioning (OzoneOP) protected the liver against ischemia/reperfusion (I/R) injury. The aim of this study was to investigate the role of A(1) adenosine receptor on the protective actions conferred by OzoneOP in hepatic I/R. By using a specific agonist and antagonist of the A(1) subtype receptor (2-chloro N6 cyclopentyladenosine, CCPA and 8-cyclopentyl-1,3-dipropylxanthine, DPCPX respectively), we studied the role of A(1) receptor in the protective effects of OzoneOP on the liver damage, nitiric oxide (NO) generation, adenosine deaminase activity and preservation of the cellular redox balance. Immunohistochemical analysis of nuclear factor-kappa B (NF-kappaB), tumor necrosis factor alpha (TNF-alpha) and heat shock protein-70 (HSP-70) was performed. OzoneOP prevented and/or ameliorated ischemic damage. CCPA showed a similar effect to OzoneOP + I/R group. A(1)AR antagonist DPCPX blocked the protective effect of OzoneOP. OzoneOP largely reduced the intensity of the p65 expression, diminished TNF-alpha production, and promoted a reduction in HSP-70 immunoreactivity. In summary, OzoneOP exerted protective effects against liver I/R injury through activation of A(1) adenosine receptors (A(1)AR). Adenosine and (.)NO produced by OzoneOP may play a role in the pathways of cellular signalling which promote preservation of the cellular redox balance, mitochondrial function, glutathione pools as well as the regulation of NF-kappaB and HSP-70.

Ozonoterapia combinada en el tratamiento del paciente portador de hernia discal lumbar: estudio preliminar / Combined ozone therapy in the treatment of the patient with lumbar herniated disc: preliminary study

Se estudió el efecto de la ozonoterapia por vía rectal e intramuscular (paravertebral) sobre variables clínicas y hallazgos imagenológicos en pacientes portadores de hernia discal lumbar. Es conocido que la hernia discal lumbar es causa de sacrolumbalgia y en muchos de los casos llega a invalidar al paciente, constituyendo un problema de salud importante en el mundo actual. A pesar de que existen diferentes esquemas de tratamientos (conservadores y quirúrgicos) para abordar el paciente con hernia discal, muchas veces estos fracasan. Teniendo en cuenta, por otra parte, las propiedades de la ozonoterapia sobre el metabolismo del oxígeno y sobre mediadores de la inflamación, fue objetivo de este trabajo valorar el efecto de la ozonoterapia sobre la calidad de vida en pacientes con hernia discal lumbar. Para desarrollar este estudio se utilizó una muestra de 37 pacientes portadores de hernia discal lumbar, a estos se les midió por escala la fuerza muscular y el grado de reflectividad osteotendinosa, así como la intensidad del dolor antes y al finalizar el tratamiento con ozono, además se realizaron estudios imagenológicos antes y después de haber concluido el esquema terapéutico. Los resultados mostraron que la terapia con ozono reflejo un efecto beneficioso en las variables clínicas estudiadas en estos pacientes además de incidir significativamente en el mejoramiento desde el punto de vista imagenológico(AU)

Ozonoterapia combinada en el tratamiento del paciente portador de hernia discal lumbar: estudio preliminar / Combined ozone therapy in the treatment of the patient with lumbar herniated disc: preliminary study

Se estudió el efecto de la ozonoterapia por vía rectal e intramuscular (paravertebral) sobre variables clínicas y hallazgos imagenológicos en pacientes portadores de hernia discal lumbar. Es conocido que la hernia discal lumbar es causa de sacrolumbalgia y en muchos de los casos llega a invalidar al paciente, constituyendo un problema de salud importante en el mundo actual. A pesar de que existen diferentes esquemas de tratamientos (conservadores y quirúrgicos) para abordar el paciente con hernia discal, muchas veces estos fracasan. Teniendo en cuenta, por otra parte, las propiedades de la ozonoterapia sobre el metabolismo del oxígeno y sobre mediadores de la inflamación, fue objetivo de este trabajo valorar el efecto de la ozonoterapia sobre la calidad de vida en pacientes con hernia discal lumbar. Para desarrollar este estudio se utilizó una muestra de 37 pacientes portadores de hernia discal lumbar, a estos se les midió por escala la fuerza muscular y el grado de reflectividad osteotendinosa, así como la intensidad del dolor antes y al finalizar el tratamiento con ozono, además se realizaron estudios imagenológicos antes y después de haber concluido el esquema terapéutico. Los resultados mostraron que la terapia con ozono reflejo un efecto beneficioso en las variables clínicas estudiadas en estos pacientes además de incidir significativamente en el mejoramiento desde el punto de vista imagenológico.

Therapeutic efficacy of ozone in patients with diabetic foot.

Oxidative stress is suggested to have an important role in the development of complications in diabetes. Because ozone therapy can activate the antioxidant system, influencing the level of glycemia and some markers of endothelial cell damage, the aim of this study was to investigate the therapeutic efficacy of ozone in the treatment of patients with type 2 diabetes and diabetic feet and to compare ozone with antibiotic therapy. A randomized controlled clinical trial was performed with 101 patients divided into two groups: one (n = 52) treated with ozone (local and rectal insufflation of the gas) and the other (n = 49) treated with topical and systemic antibiotics. The efficacy of the treatments was evaluated by comparing the glycemic index, the area and perimeter of the lesions and biochemical markers of oxidative stress and endothelial damage in both groups after 20 days of treatment. Ozone treatment improved glycemic control, prevented oxidative stress, normalized levels of organic peroxides, and activated superoxide dismutase. The pharmacodynamic effect of ozone in the treatment of patients with neuroinfectious diabetic foot can be ascribed to the possibility of it being a superoxide scavenger. Superoxide is considered a link between the four metabolic routes associated with diabetes pathology and its complications. Furthermore, the healing of the lesions improved, resulting in fewer amputations than in control group. There were no side effects. These results show that medical ozone treatment could be an alternative therapy in the treatment of diabetes and its complications.

Ozone therapy on rats submitted to subtotal nephrectomy: role of antioxidant system.

Chronic renal failure (CRF) represents a world health problem. Ozone increases the endogenous antioxidant defense system, preserving the cell redox state. The aim of this study is to evaluate the effect of ozone/oxygen mixture in the renal function, morphology, and biochemical parameters, in an experimental model of CRF (subtotal nephrectomy). Ozone/oxygen mixture was applied daily, by rectal insufflation (0.5 mg/kg) for 15 sessions after the nephrectomy. Renal function was evaluated, as well as different biochemical parameters, at the beginning and at the end of the study (10 weeks). Renal plasmatic flow (RPF), glomerular filtration rate (GFR), the urine excretion index, and the sodium and potassium excretions (as a measurement of tubular function) in the ozone group were similar to those in Sham group. Nevertheless, nephrectomized rats without ozone (positive control group) showed the lowest RPF, GFR, and urine excretion figures, as well as tubular function. Animals treated with ozone showed systolic arterial pressure (SAP) figures lower than those in the positive control group, but higher values compared to Sham group. Serum creatinine values and protein excretion in 24 hours in the ozone group were decreased compared with nephrectomized rats, but were still higher than normal values. Histological study demonstrated that animals treated with ozone showed less number of lesions in comparison with nephrectomized rats. Thiobarbituric acid reactive substances were significantly increased in nephrectomized and ozone-treated nephrectomized rats in comparison with Sham group. In the positive control group, superoxide dismutase (SOD) and catalase (CAT) showed the lowest figures in comparison with the other groups. However, ozone/oxygen mixture induced a significant stimulation in the enzymatic activity of CAT, SOD, and glutathione peroxidase, as well as reduced glutathione in relation with Sham and positive control groups. In this animal model of CRF, ozone rectal administrations produced a delay in the advance of the disease, protecting the kidneys against vascular, hemorheological, and oxidative mechanisms. This behavior suggests ozone therapy has a protective effect on renal tissue by downregulation of the oxidative stress shown in CRF.

Reversion by ozone treatment of acute nephrotoxicity induced by cisplatin in rats.

BACKGROUND: Ozone therapy has become a useful treatment for pathological processes, in which the damage mediated by reactive oxygen species is involved. Several lines of evidence suggest that cisplatin-induced acute nephrotoxicity is partially mediated by reactive oxygen species AIMS: To analyze the effect of ozone administration after cisplatin-induced acute nephrotoxicity. METHODS: Male Sprague-Dawley rats were treated with five intra-rectal applications of ozone/oxygen mixture at 0.36, 1.1 and 1.8 mg/kg after cisplatin intraperitoneal injection (6 mg/kg). Serum and kidneys were taken off 5 days after cisplatin treatment. Creatinine was measured in the serum and the activities of antioxidant enzymes and thiobarbituric acid reactive substances and glutathione content were analyzed in renal homogenate. RESULTS: Ozone treatment diminished the increase in serum creatinine levels, the glutathione depletion and also reversed the inhibition of superoxide dismutase, catalase and glutathione peroxidase activities induced by cisplatin in the rat kidney. Also, the renal content of thiobarbituric reactive substances was decreased by ozone/oxygen mixture applied after cisplatin. CONCLUSION: Intrarectal applications of ozone reversed the renal pro-oxidant unbalance induced by cisplatin treatment by the way of stimulation to some constituents of antioxidant system in the kidney, and thereby it decreased the renal damage.

Aplicación de la ozonoterapia en el tratamiento de conductos infectados / Application of ozonotherapy in the tratament of infected radicular ducts

Se evalúa el poder germicida del aceite ozonizado (Oleozón) en el tratamiento de conductos radiculares infectados. La muestra fue de 200 pacientes adultos, con diagnóstico de área de rarefacción periapical radiolúcidas, con presencia de fístula o no en dientes monorradiculares. La selección de los pacientes, a ambos grupos de estudio, fue aleatoria. El grupo experimental recibió curas con Oleozón, impregnado en bolilla de algodón estéril, en cámara a la entrada de los conductos. El cambio de cura se realizó cada 48 horas. En el grupo se aplicó en el mismo lugar y de forma similar un bactericida líquido (Cresophene). En este grupo el cambio de cura se realizó a los 7 días. Además, se les realizaron exámenes clínicos y radiológico al inicio y al final del tratamiento. Los criterios de evaluación que se tuvieron en cueta fueron : ausencia o presencia de dolor y conducto seco o húmedo. En el grupo tratado con Oleozón se obtuvo el 91 por ciento de curados en comparación con el grupo control que sólo alcanzó el 55 por ciento con una diferencia altamente significativa (p < 0,01). El alto poder germicida del Oleozón se demostró no sólo en obtener un mayor tanto por ciento de curación , al compararlo con un grupo con tratamiento convencional sino en la rapidez que se alcanzaba esta curación. En ninguno de los pacientes tratados se observó intolerancia al Oleozón (AU)

Estudio teratogénico del ozono administrado por enema a ratas Wistar / Teratogenic study of ozone adinistered by enema to Wistar rats

Se estudia el efecto del ozono en ratas Wistar gestantes. El productó se administró por enema desde el sexto hasta el decimoquinto día de la gestación. No hubo efectos teratogénicos ni ningún otro tipo de toxicidad embrionaria. En las condiciones de este experimento el ozono no tuvo efectos adversos sobre el desarrollo en el útero

Estudio teratogénico del ozono administrado por enema a ratas Wistar / Teratogenic study of ozone administered by enema to Wistar rats

Se estudia el efecto del ozono en ratas Wistar gestantes. El productó se administró por enema desde el sexto hasta el decimoquinto día de la gestación. No hubo efectos teratogénicos ni ningún otro tipo de toxicidad embrionaria. En las condiciones de este experimento el ozono no tuvo efectos adversos sobre el desarrollo en el útero