Response of cultured normal canine mammary epithelial cells to deracoxib—doxorubicin combination

Currently, there is a growing interest in combining anticancer drugs with the aim to improve outcome in patients suffering from tumours and reduce the long-term toxicity associated with the current standard of treatment. In this study, we evaluated the possible role of deracoxib against the toxicity of doxorubicin on normal canine mammary epithelial cells. The effect of deracoxib and doxorubicin combination on cell viability was determined by MTT assay. Apoptosis was characterised by flow cytometry. Cell nitrite concentrations were measured with the Griess reaction. Deracoxib (50 and 100 μM) treatment decreased the cytotoxic action of doxorubicin at 0.9 μM in the cells, from 33.63% to 13.4% and 25.82%, respectively. Our results also showed that the reverse effect of deracoxib on doxorubicin-induced cytotoxic activity in the cells was associated with a marked (3.04- to 3.57-fold) decrease in apoptosis. In additional studies identifying the mechanism of the observed effect, deracoxib exhibited an activity to prevent doxorubicin-mediated overproduction of nitric oxide in the cells. Our in vitro study results indicate that deracoxib (50 and 100 μM) can be beneficial in protecting normal cells from the toxic effect of doxorubicin in conjunction with apoptosis by the modulation of nitric oxide production

LACK OF THE ANTIMUTAGENIC EFFECT OF ASCORBIC ACID ON THE GENOTOXICITY OF ALBENDAZOLE IN MOUSE BONE MARROW CELLS

The present study was designed to evaluate the genotoxicity of albendazole (ABZ) in mice using the micronucleus test. Mice were treated by gavage with 500, 1,000, and 1,500 mg of ABZ kg(-1) b.w., which corresponds to 1/6, 1/3, and 1/2 of the oral LD50 of ABZ. In the second part of the study, the possible protective role of vitamin C (vit. C) was investigated against the genotoxic effect of ABZ. The mice received 200 mg of vit C kg(-1) b.w. simutaneously with ABZ. Bone marrow samples were taken 48 h after the treatment. ABZ induced a statistically significant increase (P<0.001) in the percentage of micronucleaoted polychromatic erythrocytes (MNPCE) in the 1,000 and 1,500 mg of ABZ kg(-1) b.w. treatment groups when compared with the negative control. On the other hand, the percentage of induced MNPCE was reduced at various levels in all ABZ treated groups simultaneously treated with an oral administration of vit. C. There were no statistically significant differences in the MNPCE frequency of these groups. The results of the study indicated that ABZ is a potential genotoxic agent, and that no protective effects of vit. C were observed against the genotoxicity of ABZ

The fate of oxytetracycline in two-phase and single-phase anaerobic cattle manure digesters and its effects on microbial communities

BACKGROUND: The fate of veterinary antibiotics in manure digesters is of great concern because of poor biomethane recovery and potential development of antibiotic resistant bacteria in the environment

Development of a new catalase activity assay for biological samples using optical CUPRAC sensor

novel catalase activity assay was developed for biological samples (liver and kidney tissue homogenates) using a rapid and low-cost optical sensor-based 'cupric reducing antioxidant capacity' (CUPRAC) method. The reagent, copper(II)-neocuproine (Cu(II)-Nc) complex, was immobilized onto a cation-exchanger film of Nafion, and the absorbance changes associated with the formation of the highly-colored Cu(I)-Nc chelate as a result of reaction with hydrogen peroxide (H2O2) was measured at 450 nm. When catalase was absent, H2O2 produced the CUPRAC chromophore, whereas catalase, being an effective H2O2 scavenger, completely annihilated the CUPRAC signal due to H2O2. Thus, the CUPRAC absorbance due to H2O2 oxidation concomitant with Cu(I)-Nc formation decreased proportionally with catalase. The developed sensor gave a linear response over a wide concentration range of (0.68-78.6 mu M). This optical sensor-based method applicable to tissue homogenates proved to be efficient for low hydrogen peroxide concentrations (physiological and nontoxic levels) to which the widely used UV method is not accurately responsive. Thus, conventional problems of the UV method arising from relatively low sensitivity and selectivity, and absorbance disturbance due to gaseous oxygen evolution were overcome. The catalase findings of the proposed method for tissue homogenates were statistically alike with those of HPLC. (C) 2014 Elsevier B.V. All rights reserved

Strategies of targeting the extracellular domain of RON tyrosine kinase receptor for cancer therapy and drug delivery

Cancer is one of the most important life-threatening diseases in the world. The current efforts to combat cancer are being focused on molecular-targeted therapies. The main purpose of such approaches is based on targeting cancer cell-specific molecules to minimize toxicity for the normal cells. RON (Recepteur d'Origine Nantais) tyrosine kinase receptor is one of the promising targets in cancer-targeted therapy and drug delivery

Characterizing the Hot Spots Involved in RON-MSP beta Complex Formation Using In Silico Alanine Scanning Mutagenesis and Molecular Dynamics Simulation

Purpose: Implication of protein-protein interactions (PPIs) in development of many diseases such as cancer makes them attractive for therapeutic intervention and rational drug design. RON (Recepteur d'Origine Nantais) tyrosine kinase receptor has gained considerable attention as promising target in cancer therapy. The activation of RON via its ligand, macrophage stimulation protein (MSP) is the most common mechanism of activation for this receptor. The aim of the current study was to perform in silico alanine scanning mutagenesis and to calculate binding energy for prediction of hot spots in proteinprotein interface between RON and MSP beta chain (MSP beta)

Identification of a RON tyrosine kinase receptor binding peptide using phage display technique and computational modeling of its binding mode

RON (Recepteur d'Origine Nantais) tyrosine kinase receptor is a promising target for therapeutic intervention in cancer therapy. The aim of this work was identification of RON-binding peptides using phage display and computational modeling their mode of binding. A 12-mer peptide phage library was utilized to perform biopanning against RON. The RON-binding ability of the selected peptide-displaying phage and their possible binding sites were examined by ELISA. Binding modes and affinities were also predicted by docking and molecular dynamics (MD) simulation. The results of ELISA experiment showed that P6 peptide displaying phage has higher affinity for RON compared to others and its binding site is located out of ligand binding site. Docking and MD simulation results also indicated higher affinity of P6 to RON as well as its exosite-binding feature. Taken together, our data suggest a capacity for P6 peptide (FEHSLYKEMTHL) to be utilized as RON binding agent, and hence be used for various purposes, including design of drug delivery systems for transferring cytotoxic agents to RON-positive cancer cells, interfering with RON signaling, peptidomimetics design, and diagnostic imaging

The Effects of Piroxicam and Deracoxib on Canine Mammary Tumour Cell Line

Cyclooxygenase (COX) inhibitors, already widely used for the treatment of pain and inflammation, are considered as promising compounds for the prevention and treatment of neoplasia. The aim of our study was to determine the direct antiproliferative effects of nonsteroidal anti-inflammatory drugs (NSAIDs), piroxicam and deracoxib, at a variety of concentrations as both single and combined treatments on canine mammary carcinoma cell line CMT-U27 and to understand the mechanisms of cell death. MTT assay was performed to determine cell viability, and flow cytometric analyses were performed to evaluate apoptosis and cell cycle alterations. Significant decrease in cell viability was observed at high concentrations of piroxicam and deracoxib in both single and combined treatments after 72 h incubation. Combined treatment produced a significantly greater inhibition than that caused by either agent alone. Also apoptotic cell number was increased by both drugs at the cytotoxic concentrations. However, concomitant treatment of cells with piroxicam and deracoxib resulted in significant induction of apoptosis at lower concentrations and accumulation of cells in the G(0)/G(1) phase. Significant cytotoxic effects exhibited by the combination of piroxicam and deracoxib against canine mammary carcinoma cells in vitro suggest an attractive approach for the treatment of canine mammary carcinoma

Investigation of the in vitro cytotoxic effects and wound healing activity of ternary composite substance (hollow silica sphere/gum arabic/methylene blue)

We aimed to test the adsorption of the methylene blue (MB) on HSEPCGUM as a dye and also to test the obtained ternary biocomposite substance (HSEPCGUM-MB) on wound healing. Hollow silica spheres (HSS) are used in the pharmaceutical and biochemical field, because of low toxic, highly biocompatible and mechanically stable by large surface areas. HSS was obtained by mechanochemistry method. The obtained HSS was treated with epichlorohydrin to carry out an epoxidation process (HSEPC). Then, HSEPC was functionalized by treatment with gum arabic (HSEPCGUM). MB was adsorbed onto HSEPCGUM and the adsorption maximum capacity of HSEPCGUM (X-max) was obtained 333 mg g(-1). For in vitro studies, according to the cytotoxicity test results, ternary biocomposite substance (HSEPCGUM-MB) was studied at non-cytotoxic concentrations 10, 50 and 100 mu g/ml and wound closure was found as 55% (100 g/m1) as compared to control. (C) 2018 Elsevier B.V. All rights reserved

Protective effects of Salvia officinalis extract against cyclophosphamide-induced genotoxicity and oxidative stress in rats

Medicinal plants, especially rich in polyphenolic compounds, have been suggested to be chemopreventive on account of antioxidative properties. Salvia officinalis L., an aromatic and medicinal plant, is widely used in folk medicine and is well known for its antioxidant properties. Therefore, the present study was designed to investigate the antioxidative, possible genotoxic, and antigenotoxic potency of S. officinalis extract against cyclophosphamide (CYP)-induced oxidative stress and genotoxicity in Wistar albino rats. Animals were orally dosed with S. officinalis extract (50, 100, and 150 mg/kg body weight) for 7 days before the administration of a single intraperitoneal dose of CYP (40 mg/kg body weight). The biochemical and cytogenetic determinations were carried out 24 h aft er CYP injection. The activities of malondialdehyde, superoxide dismutase, and catalase were determined in liver, kidney, and heart tissues. The frequency of micronucleated polychromatic erythrocytes (MNPCEs) and the ratio of polychromatic erythrocytes to normochromatic erythrocytes (PCE/NCE) were also evaluated. Additionally, the antioxidant capacity of the extract was analyzed. The micronucleus assay revealed that all doses of the extract tested presented no genotoxic activity; in addition, the 2 highest doses reduced the MNPCEs and increased the PCE/NCE ratio in the bone marrow and restored the oxidative stress markers in CYP-treated rats. In correlation with these findings, S. officinalis extract exhibited high antioxidant capacity. The results of the present study suggest that the methanolic extract of S. officinalis has a protective effect against CYP-induced oxidative stress and genotoxicity through its antioxidant property