Erythropoetin and Erythropoetin Receptor Systems, and Oxidative Status in Patients with Basal Cell Carcinoma

Introduction: Erythropoietin (Epo) controls a variety of signal transduction pathways during oxidative stress. The main function of Epo and its receptor (EpoR) is the stimulation of erythropoiesis.&nbsp; Aim: The role of Epo and EpoR on non-hematopoietic normal and cancerous tissues is still poorly understood. This is the first report in which we aimed to investigate the role of Epo and EpoR systems at oxidative condition in human basal cell carcinoma (BCC), which is the most common tumour in the world.Materials and methods: Fresh normal and cancerous skin paired tissue was obtained from 63 patients who underwent curative BCC resection in Kahramanmaras, Turkey. Preliminary diagnosis of BCC was made in the dermatology clinic by excision and then the diagnosis was confirmed as histopathologic findings. Oxidative stress biomarkers such as superoxide dismutase (SOD) and catalase (CAT) activities, and malondialdehyde (MDA) levels in biopsy samples were measured spectrophotometrically, and also the levels of Epo and EpoR were measured by ELISA.&nbsp; Results: While the levels of MDA in cancerous tissue of patients with skin BCC were significantly higher than normal neighbouring skin tissue (p<0.05), SOD and CAT activities decreased (p<0.05). Furthermore, a remarkable increase was found in the Epo level ofpatients with skin BCC in comparison with the normal neighbouring skin tissue (p<0.05). However, we found that EpoR levelsdecreased (p<0.05).&nbsp; Conclusions: Results indicate that there is an active oxidative process in BCC biopsies. The levels of increased Epo and decreased EpoR in oxidative condition due to hypoxia may aggravate tumour growth by its angiogenic activity.

NR4A3 is oxidative stress responsible transcription factor through HMOX1, and also controls cell cycle through CCNE1 and CDK2 in pancreatic islet derived 1.1B4 cells

The mechanism of antioxidant defense system is still controversial. As islet β-cell is weak in oxidative condition, that causes diabetes mellitus, therefore, antioxidant defense system of human pancreatic islet derived 1.1B4 cell was analyzed. Cells were exposed to H2O2 and comprehensive gene expression was analyzed by Agilent human microarray. HMOX1 and NR4A3, member of orphan receptor, were up-regulated. Therefore, NR4A3 was knocked down with siRNA, then analyzed gene expression by microarray, and found that the knocked down cells were weak in oxidative stress. HMOX1 expression was strongly inhibited by siRNA of NR4A3, and NR4A3 responsible sequence of aaggtca was found near the HMOX1 gene, suggesting NR4A3 is oxidative stress responsible transcription factor through HMOX1 expression. The expression of CCNE1 and CDK2 was also inhibited by knocked down of NR4A3, it is suggested NR4A3 is also important transcription factor for cell growth regulation.

Use of acetylsalicylic acid and agronomic performance of potatoes in Lima region.

Background: Salicylic acid participates in the regulation of the plant's response to a series of environmental stresses such as extreme temperatures, salinity, and oxidative condition of potato production, so it is necessary to determine a safe application dosage for potato in field conditions. Methods: The purpose of the research was to evaluate the effect of increasing doses of acetylsalicylic acid on the agronomic traits of two potato cultivars in Lima, Peru. In a field experiment, an RCBD with 3 replications was used under a split-plot arrangement in which five doses (0.0, 0.2, 0.4, 0.6, and 0.8 mM) of acetylsalicylic acid plus control with Biol (biostimulant) were assigned to plots, and two potato cultivars ('Perricholi' and 'Única') were assigned to subplots. The average temperatures were between 15 and 21°C, the air relative humidity between 61 and 73% and the soil was loam textural class, free of salts. Results: Statistical differences (p <0.05) were found between the two potato cultivars for most of the characters studied. The effects of doses of acetylsalicylic acid showed statistical differences for sprouting, vegetative vigor, and percentage of flowering; likewise, significant interactions (p <0.05) were shown between potato cultivars and dose of acetylsalicylic acid for the number of stems per plant and percentage of flowering, which would indicate a specific effect for the dose of acetylsalicylic acid in each potato cultivar. Conclusion: No dose of the product significantly affected the potato yield in two potato cultivars however, the effect of the dosage of acetylsalicylic acid improved the performance of the crop in terms of sprouting in the field, vegetative vigor, a greater number of stems per plant as well as stimulation of flowering, with respect to control.

MiR-599 Protects Cardiomyocytes against Oxidative Stress-Induced Pyroptosis

Oxidative stress is a crucial factor and key promoter of a variety of cardiovascular diseases associated with cardiomyocyte injury. Emerging literatures suggest that pyroptosis plays a key role in cardiac damages. However, whether pyroptosis contributes to cardiomyocyte injury under oxidative stress and the underlying molecular mechanisms are totally unclear. This study was designed to investigate the potential role of pyroptosis in H2O2-induced cardiomyocyte injury and to elucidate the potential mechanisms. Primary cardiomyocytes from neonatal Wistar rats were utilized. These myocytes were treated with different concentrations of H2O2 (25, 50, and 100 μM) for 24 h to induce oxidative injury. Our results indicated that mRNA and protein levels of ASC were remarkably upregulated and caspase-1 was activated. Moreover, the expressions of inflammatory factors IL-1β and IL-18 were also increased. Luciferase assay showed that miR-599 inhibited ASC expression through complementary binding with its 3 ′ UTR. MiR-599 expression was substantially reduced in H2O2-treated cardiomyocytes. Upregulation of miR-599 inhibited cardiomyocyte pyroptosis under oxidative stress, and opposite results were found by decreasing the expression of miR-599. Consistently, miR-599 overexpression ameliorated cardiomyocyte injury caused by H2O2. Therefore, miR-599 could be a promising therapeutic approach for the management of cardiac injury under oxidative condition.

Basic Pharmacological Characterization of EV-34, a New H2S-Releasing Ibuprofen Derivative

Background: Cardioprotective effects of H2S are being suggested by numerous studies. Furthermore, H2S plays a role in relaxation of vascular smooth muscle, protects against oxidative stress, and modulates inflammation. Long-term high-dose use of NSAIDs, such as ibuprofen, have been associated with enhanced cardiovascular risk. The goal of the present work is the synthesis and basic pharmacological characterization of a newly designed H2S-releasing ibuprofen derivative. Methods: Following the synthesis of EV-34, a new H2S-releasing derivative of ibuprofen, oxidative stability assays were performed (Fenton and porphyrin assays). Furthermore, stability of the molecule was studied in rat serum and liver lysates. H2S-releasing ability of the EC-34 was studied with a hydrogen sulfide sensor. MTT (3-(4,5-dimethylthiazol 2-yl)-2,5-(diphenyltetrazolium bromide)) assay was carried out to monitor the possible cytotoxic effect of the compound. Cyclooxygenase (COX) inhibitory property of EV-34 was also evaluated. Carrageenan assay was carried out to compare the anti-inflammatory effect of EV-34 to ibuprofen in rat paws. Results: The results revealed that the molecule is stable under oxidative condition of Fenton reaction. However, EV-34 undergoes biodegradation in rat serum and liver lysates. In cell culture medium H2S is being released from EV-34. No cytotoxic effect was observed at concentrations of 10, 100, 500 µM. The COX-1 and COX-2 inhibitory effects of the molecule are comparable to those of ibuprofen. Furthermore, based on the carrageenan assay, EV-34 exhibits the same anti-inflammatory effect to that of equimolar amount of ibuprofen (100 mg/bwkg). Conclusion: The results indicate that EV-34 is a safe H2S releasing ibuprofen derivative bearing anti-inflammatory properties.

Degradation Studies of Lacidipine: Identification, Isolation and Structural Characterization of Stress Degradation Products using LCMS, Mass mediated Prep-HPLC, NMR and HRMS

The stability of lacidipine drug under stress conditions and the identification of the degradation products, according to ICH guidelines Q1A (R2) were investigated in the hydrolytic and oxidative stress conditions. The drug degradation occurred under hydrolytic conditions like (acidic and basic) while it was stable in the oxidative condition. Three degradation products were formed under acidic condition and one degradation product was formed under basic condition, which was separated by using APMS (Auto Purification Mass Spectrometer) and gradient elution with C18 column. The four degradants have not been characterized earlier and in the present study all the structures were established and characterized using NMR spectroscopy (1D and 2D) and HRMS (high resolution mass spectrometer).

Validated stability indicating assay method for tenofovir implementing design of experiment approach for forced degradation

Tenofovir, an antiretroviral drug known as nucleoside analogue reverse transcriptase inhibitor, was evaluated for its degradation behavior. The study involved optimization of forced degradation study using the design of experiment (DoE) approach and validation of developed stability indicating assay method (SIAM) for tenofovir. Chromatographic separation was achieved on Kromasil C18 column using mixture of methanol and ammonium acetate buffer pH 8.5 as mobile phase in isocratic elution mode. Detection was done using PDA detector. Tenofovir showed degradation at alkali, acid hydrolysis and oxidative condition while it was stable to photolysis and thermal stress condition. The optimization of forced degradation conditions, like hydrolytic and oxidative, was done by application 3n full factorial designs, which helped to obtain desired drug degradation. The proposed method was successfully validated as per ICH Q2 (R1) guidelines.