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Cellular and Molecular Biology... Feb 2022Laccase producing fungus Pleurotus floridanus was isolated from Siruvani forest, Tamil Nadu, India. The potential of P. floridanus to produce laccase by using various...
Laccase producing fungus Pleurotus floridanus was isolated from Siruvani forest, Tamil Nadu, India. The potential of P. floridanus to produce laccase by using various lignocellulosic substrates was screened under submerged fermentation. Laccase production in the presence of lignocellulosic substrates such as rice, wheat and maize bran as a sole source of carbon as well as an additional supplement was examined. Laccase activity of P. floridanus using varied substrates was observed in the order of rice bran > wheat bran > maize bran. The isolate showed maximum laccase activity of 13.29±0.01 U/mL using rice bran as a carbon source within 11 days. This was 18 fold higher than the control media that lacks lignocellulosic substrates. The diclofenac tolerance was assessed in solid media at various concentrations and the results showed that the mycelia growth is not significantly affected by the drug. Finally, the laccase mediated degradation of diclofenac at a concentration of 10 mg/L showed 98% degradation in 2 h. The phytotoxicity of the crude laccase treated diclofenac was lower than the untreated diclofenac. In conclusion, findings suggested direct application of crude laccase produced from P. floridanus using agro-residues as ideal substrate for environmental applications.
Topics: Biotransformation; Carbon; Diclofenac; India; Laccase; Pleurotus
PubMed: 35818224
DOI: 10.14715/cmb/2021.67.5.56 -
Anticancer Research Apr 2022The anticancer potential of indomethacin and diclofenac has been reported against several types of cancer cells. In this study, we investigated the enhancement effect of...
BACKGROUND/AIM
The anticancer potential of indomethacin and diclofenac has been reported against several types of cancer cells. In this study, we investigated the enhancement effect of a coumaric acid derivative found in some Piper plants, N-trans-p-coumaroyltyramine (TCT) on indomethacin and diclofenac cytotoxicity in breast cancer cells.
MATERIALS AND METHODS
MCF-7 and mitoxantrone-resistant MCF-7 (MCF-7/MX) cancer cells were treated with indomethacin or diclofenac in the presence of TCT for 48 h. Cell viability, apoptosis, mitochondrial function and signaling proteins were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Hoechst 33342, tetramethyl-rhodamine-ethyl-ester and western blot analysis, respectively.
RESULTS
Combination treatment resulted in significant reduction of cell viability and mitochondrial membrane potential, whereas the ratio of BCL2-associated X, apoptosis regulator to BCL2 apoptosis regulator, and apoptosis increased. The enhancing effect of TCT was related to reduced nuclear factor-erythroid factor 2-related factor 2/heme oxygenase-1 expression, and increased activation of the protein kinase RNA-like endoplasmic reticulum kinase/eukaryotic initiation factor 2 alpha/activating transcription factor 4/C/EBP homologous protein signaling pathways.
CONCLUSION
TCT in combination with indomethacin or diclofenac promoted endoplasmic reticulum stress-dependent apoptosis in breast cancer cells.
Topics: Apoptosis; Coumaric Acids; Diclofenac; Endoplasmic Reticulum Stress; Humans; Indomethacin; MCF-7 Cells
PubMed: 35347001
DOI: 10.21873/anticanres.15659 -
Analytical Methods : Advancing Methods... May 2024The non-steroidal anti-inflammatory drug (NSAID) diclofenac (DCF) is an important environmental contaminant occurring in surface waters all over the world, because,...
The non-steroidal anti-inflammatory drug (NSAID) diclofenac (DCF) is an important environmental contaminant occurring in surface waters all over the world, because, after excretion, it is not adequately removed from wastewater in sewage treatment plants. To be able to monitor this pollutant, highly efficient analytical methods are needed, including immunoassays. In a medical research project, monoclonal antibodies against diclofenac and its metabolites had been produced. Based on this monoclonal anti-DCF antibody, a new indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed and applied for environmental samples. The introduction of a spacer between diclofenac and the carrier protein in the coating conjugate led to higher sensitivity. With a test midpoint of 3 μg L and a measurement range of 1-30 μg L, the system is not sensitive enough for direct analysis of surface water. However, this assay is quite robust against matrix influences and can be used for wastewater. Without adjustment of the calibration, organic solvents up to 5%, natural organic matter (NOM) up to 10 mg L, humic acids up to 2.5 mg L, and salt concentrations up to 6 g L NaCl and 75 mg L CaCl are tolerated. The antibody is also stable in a pH range from 3 to 12. Cross-reactivity (CR) of 1% or less was determined for the metabolites 4'-hydroxydiclofenac (4'-OH-DCF), 5-hydroxydiclofenac (5-OH-DCF), DCF lactam, and other NSAIDs. Relevant cross-reactivity occurred only with an amide derivative of DCF, 6-aminohexanoic acid (DCF-Ahx), aceclofenac (ACF) and DCF methyl ester (DCF-Me) with 150%, 61% and 44%, respectively. These substances, however, have not been found in samples. Only DCF-acyl glucuronide with a cross-reactivity of 57% is of some relevance. For the first time, photodegradation products were tested for cross-reactivity. With the ELISA based on this antibody, water samples were analysed. In sewage treatment plant effluents, concentrations in the range of 1.9-5.2 μg L were determined directly, with recoveries compared to HPLC-MS/MS averaging 136%. Concentrations in lakes ranged from 3 to 4.4 ng L and were, after pre-concentration, determined with an average recovery of 100%.
Topics: Diclofenac; Antibodies, Monoclonal; Water Pollutants, Chemical; Enzyme-Linked Immunosorbent Assay; Anti-Inflammatory Agents, Non-Steroidal; Environmental Monitoring; Wastewater
PubMed: 38742423
DOI: 10.1039/d3ay01333b -
European Journal of Anaesthesiology Mar 2021Diclofenac and other NSAIDs are routinely used in the postoperative period. Their effect on fracture healing remains unclear and controversial.
BACKGROUND
Diclofenac and other NSAIDs are routinely used in the postoperative period. Their effect on fracture healing remains unclear and controversial.
OBJECTIVE
The primary outcome was to assess the potential cytotoxicity of clinically relevant concentrations of diclofenac on human osteoblasts.
DESIGN
Laboratory in vitro study.
SETTING
Institute of Physiology, Zurich, Center for Integrative Human Physiology, University of Zurich.
MATERIALS
Monolayers of human osteoblasts.
INTERVENTIONS
Exposure of human osteoblast monolayers to several concentrations of diclofenac, for different periods of time, with and without an artificially induced inflammatory process.
MAIN OUTCOME MEASURES
Cell count, cell viability, cell proliferation and apoptosis.
RESULTS
A concentration-mediated, time and exposure dependent cytotoxic effect of diclofenac-mediated apoptosis was observed. Stimulated inflammatory conditions seemed to reduce toxic effects.
CONCLUSION
Cytotoxic effects of diclofenac are exposure, time and concentration dependent. Simulating aspects of inflammatory conditions seems to increase resistance to diclofenac cytotoxicity, especially in the presence of higher concentration and longer exposure time.
Topics: Anti-Inflammatory Agents, Non-Steroidal; Cell Proliferation; Diclofenac; Humans; Inflammation; Osteoblasts
PubMed: 33122572
DOI: 10.1097/EJA.0000000000001363 -
Journal of the Science of Food and... Aug 2024Seafood consumers are widely exposed to diclofenac due to the high contamination levels often present in aquatic organisms. It is a potential risk to public health due...
BACKGROUND
Seafood consumers are widely exposed to diclofenac due to the high contamination levels often present in aquatic organisms. It is a potential risk to public health due its endocrine disruptor properties. Limited information is available about diclofenac behavior after food digestion to enable a more realistic scenario of consumer exposure. This study aimed to evaluate cooking effects on diclofenac levels, and determine diclofenac bioaccessibility by an in vitro digestion assay, using commercial fish species (seabass and white mullet) as models. The production of the main metabolite 4'-hydroxydiclofenac was also investigated. Fish hamburgers were spiked at two levels (150 and 1000 ng g) and submitted to three culinary treatments (roasting, steaming and grilling).
RESULTS
The loss of water seems to increase the diclofenac levels after cooking, except in seabass with higher levels. The high bioaccessibility of diclofenac (59.1-98.3%) observed in both fish species indicates that consumers' intestines are more susceptible to absorption, which can be worrisome depending on the level of contamination. Contamination levels did not affect the diclofenac bioaccessibility in both species. Seabass, the fattest species, exhibited a higher bioaccessibility of diclofenac compared to white mullet. Overall, cooking decreased diclofenac bioaccessibility by up to 40% in seabass and 25% in white mullet. The main metabolite 4'-hydroxydiclofenac was not detected after cooking or digestion.
CONCLUSION
Thus, consumption of cooked fish, preferentially grilled seabass and steamed or baked white mullet are more advisable. This study highlights the importance to consider bioaccessibility and cooking in hazard characterization studies. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
Topics: Diclofenac; Cooking; Animals; Digestion; Food Contamination; Seafood; Fishes; Bass; Humans; Water Pollutants, Chemical; Smegmamorpha; Models, Biological
PubMed: 38437521
DOI: 10.1002/jsfa.13430 -
Drug Delivery and Translational Research May 2023Transdermal delivery of active pharmaceutical ingredients (APIs) can be challenging, since the skin possesses a rate-limiting barrier, which may be overcome when APIs...
Transdermal delivery of active pharmaceutical ingredients (APIs) can be challenging, since the skin possesses a rate-limiting barrier, which may be overcome when APIs possess certain ideal physicochemical properties. The lack thereof would require that APIs be included in drug delivery vehicles to enhance skin permeation. Hence, diclofenac was incorporated into various drug delivery vehicles (i.e., nano-emulsions, nano-emulgels, and a colloidal suspension containing drug-loaded nanoparticles) to investigate the transdermal delivery thereof, while nano-emulsions and nano-emulgels had varying concentrations of evening primrose oil (EPO). The aim of the study was to compare the topical and transdermal diclofenac delivery from the different types of vehicles and to investigate the influence the different EPO concentrations had on diclofenac delivery. After characterization, membrane release studies were performed (to determine whether the API was successfully released from the vehicle) followed by in vitro skin diffusion studies and tape stripping (to establish whether the vehicles assisted the API in reaching the target site (transdermal delivery)). Lastly, cytotoxicity studies were conducted via methyl thiazolyl tetrazolium (MTT) and neutral red (NR) assays on human keratinocyte (HaCaT) cells. Results showed minimal cytotoxic effects at concentrations equivalent to that which had permeated through the skin, while the membrane release and in vitro skin diffusion studies indicated that the nano-emulsions and the 10% EPO vehicles increased API release and diffusion when compared to the other vehicles. However, the colloidal suspension had the highest concentrations of API within the skin. Hence, all the vehicles were non-toxic and effectively delivered diclofenac through the transdermal route.
Topics: Humans; Diclofenac; Skin Absorption; Administration, Cutaneous; Skin; Emulsions; Excipients
PubMed: 36525200
DOI: 10.1007/s13346-022-01267-7 -
Current Medical Research and Opinion Jun 2019The nonsteroidal anti-inflammatory drug (NSAID) diclofenac epolamine (DHEP) formulated as a topical patch has demonstrated efficacy and safety in the localized treatment... (Review)
Review
OBJECTIVE
The nonsteroidal anti-inflammatory drug (NSAID) diclofenac epolamine (DHEP) formulated as a topical patch has demonstrated efficacy and safety in the localized treatment of acute pain from minor strains, sprains and contusions, and for epicondylitis and knee osteoarthritis. The glycosaminoglycan heparin enhances the activity of topical NSAIDs formulated as a medicated plaster, even in the absence of any significant release of heparin. Therefore, DHEP plus, a new formulation of the DHEP medicated plaster containing a small amount of heparin sodium as excipient, has been developed.
METHODS
We reviewed the pivotal and supportive studies of the clinical development program of the new patch and evaluated the role of heparin as an enhancer in the treatment of localized pain/inflammation of musculoskeletal structures, associated with post-traumatic and/or rheumatic conditions.
RESULTS
The data was consistent with the concept that heparin increased the clinical activity of the DHEP plus medicated plaster versus the reference DHEP medicated plaster through improved bioavailability due to enhanced movement of diclofenac from the plaster. Both DHEP formulations have the same dissolution profile, indicating that heparin does not change the physical and chemical characteristics of the plaster. Permeation testing showed that heparin is not released from the DHEP plus medicated plaster. Efficacy studies showed that the DHEP plus medicated plaster was significantly more effective in reducing pain than the reference marketed DHEP medicated plaster.
CONCLUSIONS
The benefit/risk assessment of DHEP plus 180 mg medicated plaster is favorable, with a safety profile equal to placebo and improved efficacy over the reference marketed DHEP medicated plaster.
Topics: Anti-Inflammatory Agents, Non-Steroidal; Diclofenac; Heparin; Humans; Inflammation; Osteoarthritis, Knee; Pain
PubMed: 30474433
DOI: 10.1080/03007995.2018.1551194 -
Behavioural Pharmacology Dec 2022Pain is a major problem that burdens the health and economy of societies worldwide. Nonsteroidal anti-inflammatory drugs (NSAIDs) are over-the-counter medications that...
Preclinical comparison of antinociceptive effects between ibuprofen, diclofenac, naproxen, and acetaminophen on acid-stimulated body stretching and acid-depressed feeding behaviors in rats.
Pain is a major problem that burdens the health and economy of societies worldwide. Nonsteroidal anti-inflammatory drugs (NSAIDs) are over-the-counter medications that are widely indicated for mild to moderate pain conditions. Clinically, the selection of a medication among this class is mainly based according to both patient's and doctor's previous experiences. Herein, we studied differences in therapeutic efficacies among the most commonly prescribed NSAIDs and acetaminophen in inflammatory pain rat model. Body stretching and food consumption behaviors were assessed after intraperitoneal administration of lactic acid. Initially, different concentrations of lactic acid were evaluated in adult male rats in both behavioral models. Acid concentrations of 1.8 and 3.2% were selected to assess the effects of ibuprofen, diclofenac, naproxen, and acetaminophen in body stretching and feeding behaviors, respectively. In the feeding study, food restriction for 1-24 h prior to feeding studies was assessed at first, and 24 h was selected for further tests. Acetaminophen (100 mg/kg), diclofenac (10 mg/kg), ibuprofen (10-32 mg/kg), and naproxen (3.2-10 mg/kg) significantly decreased acid-stimulated body stretching. Likewise, acetaminophen (100 mg/kg), diclofenac (10 mg/kg), and ibuprofen (32 mg/kg) increased food consumption significantly after 3.2% lactic acid. There were no significant differences between different test drugs efficacies in both stretching and feeding behaviors. In conclusion, feeding behavior provides a good appraisal for pain and analgesic drugs in preclinical studies. There were comparable efficacies between all tested medications in both lactic acid-stimulated body stretching and -depressed feeding behaviors.
Topics: Male; Animals; Rats; Ibuprofen; Acetaminophen; Naproxen; Diclofenac; Anti-Inflammatory Agents, Non-Steroidal; Pain; Analgesics; Feeding Behavior; Lactic Acid
PubMed: 36256732
DOI: 10.1097/FBP.0000000000000704 -
Poplar and diclofenac pollution: A focus on physiology, oxidative stress and uptake in plant organs.The Science of the Total Environment Sep 2018Poplar plants may have an important role in the removal of pharmaceuticals from contaminated waters. In this context, plant uptake of the non-steroidal anti-inflammatory...
Poplar plants may have an important role in the removal of pharmaceuticals from contaminated waters. In this context, plant uptake of the non-steroidal anti-inflammatory drug diclofenac, as well as physiological response in terms of growth traits and stress enzymes activity was assessed in Populus alba Villafranca clone, in order to establish the effectiveness of this species against pharmaceutical active compounds pollution. This evaluation was conducted in mesocosms with 1 mg L of this pharmaceutical for a maximum period of 28 days. Root appears to be the organ with clear uptake of diclofenac (14.76 ± 2.42 ng g fresh weight after 1 day of treatment), and presence of products derived from its metabolism. Indeed, 4-OH-diclofenac metabolite was detected in root tissues, indicating diclofenac uptake and metabolism inside the plants, already after 1 day of treatment. Regarding enzyme activities, glutathione-S-transferases increased in roots after long-term exposure to diclofenac, while an increase in activity of ascorbate peroxidase and glutathione reductase was detected in short and medium-term exposure, as a result of abiotic stress caused by diclofenac. Results suggest the ability of poplar to actively participate in the removal of diclofenac from water when used for phytoremediation purpose.
Topics: Biodegradation, Environmental; Biological Transport; Diclofenac; Oxidative Stress; Plant Roots; Populus; Soil Pollutants
PubMed: 29729512
DOI: 10.1016/j.scitotenv.2018.04.355 -
Journal of the American Veterinary... Jul 2024To investigate systemic absorption and gastrointestinal (GI) adverse effects of topical ketorolac 0.5% and diclofenac 0.1% ophthalmic solutions.
OBJECTIVE
To investigate systemic absorption and gastrointestinal (GI) adverse effects of topical ketorolac 0.5% and diclofenac 0.1% ophthalmic solutions.
ANIMALS
11 healthy purpose-bred Beagles.
METHODS
Dogs were randomly assigned to receive either ketorolac (n = 6) or diclofenac (5), 1 drop in both eyes 4 times daily for 28 days. Upper GI endoscopy was performed on days 0 and 29 with mucosal lesion scores (0 to 7) assigned to each region evaluated. Plasma samples were collected on days 14, 21, and 28 for measurement of diclofenac and ketorolac using high-performance liquid chromatography-mass spectrometry.
RESULTS
GI erosions and/or ulcers developed in all ketorolac-treated dogs and 1 of 5 diclofenac-treated dogs. Post-treatment mucosal lesion score for the antrum was higher in the ketorolac group than in the diclofenac group (P = .006) but not significantly different for any other region. Post-treatment antral mucosal lesion scores were significantly related to plasma ketorolac concentrations (P < .001). Ketorolac and diclofenac were detected in the plasma at all time points (median ketorolac day 14, 191 ng/mL; day 21, 173.5 ng/mL; and day 28, 179.5 ng/mL; and median diclofenac day 14, 21.1 ng/mL; day 21, 20.6 ng/mL; day 28, 27.5 ng/mL). Vomiting and decreased appetite events were observed uncommonly and were not significantly different between treatment groups.
CLINICAL RELEVANCE
GI ulceration and erosion developed after ophthalmic administration of ketorolac and diclofenac, with higher plasma concentrations and more severe GI lesions associated with ketorolac. Clients should be alerted to this potential risk with ophthalmic use and informed to watch for systemic clinical signs that would warrant veterinary reevaluation.
Topics: Animals; Dogs; Diclofenac; Anti-Inflammatory Agents, Non-Steroidal; Ketorolac; Ophthalmic Solutions; Male; Female; Dog Diseases; Administration, Topical; Gastrointestinal Diseases
PubMed: 38579753
DOI: 10.2460/javma.23.12.0707