Presence Of Non-steroidal Anti-inflammatory Drugs Research Articles

Ketoprofen promotes the conjugative transfer of antibiotic resistance among antibiotic resistant bacteria in natural aqueous environments

The emergence and spread of antibiotic resistance in the environment pose a serious threat to global public health. It is acknowledged that non-antibiotic stresses, including disinfectants, pharmaceuticals and organic pollutants, play a crucial role in horizontal transmission of antibiotic resistance genes (ARGs). Despite the widespread presence of non-steroidal anti-inflammatory drugs (NSAIDs), notably in surface water, their contributions to the transfer of ARGs have not been systematically explored. Furthermore, previous studies have primarily concentrated on model strains to investigate whether contaminants promote the conjugative transfer of ARGs, leaving the mechanisms of ARG transmission among antibiotic resistant bacteria in natural aqueous environments under the selective pressures of non-antibiotic contaminants remains unclear. In this study, the Escherichia coli (E. coli) K12 carrying RP4 plasmid was used as the donor strain, indigenous strain Aeromonas veronii containing rifampicin resistance genes in Taihu Lake, and E. coli HB101 were used as receptor strains to establish inter-genus and intra-genus conjugative transfer systems, examining the conjugative transfer frequency under the stress of ketoprofen. The results indicated that ketoprofen accelerated the environmental spread of ARGs through several mechanisms. Ketoprofen promoted cell-to-cell contact by increasing cell surface hydrophobicity and reducing cell surface charge, thereby mitigating cell-to-cell repulsion. Furthermore, ketoprofen induced increased levels of reactive oxygen species (ROS) production, activated the DNA damage-induced response (SOS), and enhanced cell membrane permeability, facilitating ARG transmission in intra-genus and inter-genus systems. The upregulation of outer membrane proteins, oxidative stress, SOS response, mating pair formation (Mpf) system, and DNA transfer and replication (Dtr) system related genes, as well as the inhibition of global regulatory genes, all contributed to higher transfer efficiency under ketoprofen treatment. These findings served as an early warning for a comprehensive assessment of the roles of NSAIDs in the spread of antibiotic resistance in natural aqueous environments.

Decomposition of non-steroidal anti-inflammatory drugs by activated sludge supported by biopreparation in sequencing batch reactor

The presence of non-steroidal anti-inflammatory drugs in wastewater from sewage treatment plants indicates that they are not completely biodegradable. The designed biopreparation based on immobilized bacteria enables the degradation of paracetamol, ibuprofen, naproxen and diclofenac at a rate of 0.50 mg/L*day, 0.14 mg/L*day, 0.16 mg/L*day and 0.04 mg/L*day, respectively. Lower degradation of drugs in the mixture than in monosubstrate systems indicates their additive, antagonistic effect, limiting the degradative capacity of microorganisms. The biopreparation is stable for at least 6 weeks in bioreactor conditions. Biochemical parameters of activated sludge functioning showed increased oxygen demand, which was related to increased ammonia concentration caused by long-term exposure of activated sludge to drugs. Reduced metabolic activity was also observed. The preparation enables decomposing drugs and their metabolites, restoring the activated sludge's functionality. The tested biopreparation can support activated sludge in sewage treatment plants in degrading non-steroidal anti-inflammatory drugs and phenolic compounds.

Phenotypic and metabolic adaptations of Rhodococcus cerastii strain IEGM 1243 to separate and combined effects of diclofenac and ibuprofen.

The increasing use of non-steroidal anti-inflammatory drugs (NSAIDs) has raised concerns regarding their environmental impact. To address this, understanding the effects of NSAIDs on bacteria is crucial for bioremediation efforts in pharmaceutical-contaminated environments. The primary challenge in breaking down persistent compounds lies not in the biochemical pathways but in capacity of bacteria to surmount stressors. In this study, we examined the biodegradative activity, morphological and physiological changes, and ultrastructural adaptations of Rhodococcus cerastii strain IEGM 1243 when exposed to ibuprofen, diclofenac, and their mixture. Our findings revealed that R. cerastii IEGM 1243 exhibited moderate biodegradative activity towards the tested NSAIDs. Cellular respiration assay showed higher metabolic activity in the presence of NSAIDs, indicating their influence on bacterial metabolism. Furthermore, catalase activity in R. cerastii IEGM 1243 exposed to NSAIDs showed an initial decrease followed by fluctuations, with the most significant changes observed in the presence of DCF and the NSAID mixture, likely influenced by bacterial growth phases, active NSAID degradation, and the formation of multicellular aggregates, suggesting potential intercellular synergy and task distribution within the bacterial community. Morphometric analysis demonstrated alterations in size, shape, and surface roughness of cells exposed to NSAIDs, with a decrease in surface area and volume, and an increase in surface area-to-volume ratio (SA/V). Moreover, for the first time, transmission electron microscopy confirmed the presence of lipid inclusions, polyphosphates, and intracellular membrane-like structures in the ibuprofen-treated cells. These results provide valuable insights into the adaptive responses of R. cerastii IEGM 1243 to NSAIDs, shedding light on the possible interaction between bacteria and pharmaceutical compounds in the environment.

Effect of nonsteroidal anti-inflammatory drugs on pelvic floor muscle regeneration in a preclinical birth injury rat model

BackgroundPelvic floor muscle (PFM) injury is a common consequence of vaginal childbirth. Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used postpartum analgesics. Multiple studies report negative effects of these drugs on limb muscle regeneration. The impact of NSAIDs on PFM recovery following birth injury has not been explored. ObjectiveUsing the validated rat model, we assessed the effects of NSAIDs on acute and longer-term PFM recovery following simulated birth injury (SBI). Study DesignThree-month old Sprague-Dawley rats were randomly assigned to (1) controls; (2) SBI; (3) SBI + NSAID; or (4) NSAID group. SBI was induced using a well-established vaginal balloon distention protocol. Ibuprofen was administered in drinking water (0.2 mg/ml) ad lib. Animals were sacrificed at 1,3,5,7,10,28 days post birth injury/ibuprofen administration. The pubocaudalis portion of the rat levator ani, which, like the human pubococcygeus, experiences larger parturition-associated strains, was harvested (N=3-9/timepoint/group). The cross-sectional areas of regenerating (embryonic myosin heavy chain (eMyHC)+) and mature myofibers were assessed at the acute and 28-day timepoints, respectively. The intramuscular collagen content was assessed at the 28-day timepoint. Myogenesis was evaluated using anti-Pax7 and anti-myogenin antibodies to identify activated and differentiated muscle stem cells, respectively. The overall immune infiltrate was assessed using anti-CD45 antibody. Expression of genes coding for pro- and anti-inflammatory cytokines was assessed by qRT-PCR at 3,5,10 days post-injury. ResultsThe pubocaudalis fiber size was significantly smaller in the SBI+NSAID compared to SBI group at 28 days post-injury, P<0.0001. The median size of eMyHC+ fibers was also significantly reduced, with fiber area distribution enriched with smaller fibers in SBI+NSAID compared to SBI group at 3 days post-injury (P<0.0001), suggesting a delay in the onset of regeneration in the presence of NSAIDs. By 10 days post-injury, the median eMyHC+ fiber size in the SBI group decreased from 7 days post-injury (P<0.0001) with a tight cross-sectional area distribution, indicating nearing completion of this state of regeneration. However, in the SBI+NSAID group, the size of eMyHC+ fibers continued to increase (P<0.0001) with expansion of the cross-sectional area distribution, signifying a delay in regeneration in these animals. NSAID decreased the muscle stem cell pool at 7 days post-injury (P<0.0001) and delayed muscle stem cell differentiation, as indicated by persistently elevated number of myogenin+ cells 7 days post-injury (P<0.05). In contrast, the proportion of myogenin+ cells returned to baseline by 5 days post-injury in the SBI group. The analysis of expression of genes coding for pro- and anti-inflammatory cytokines demonstrated only transient elevation of Tgfb1 in SBI+NSAID group at 5 but not at 10 days post-injury. Consistent with previous studies, SBI+NSAID resulted in increased deposition of intramuscular collagen compared to uninjured animals. There were no significant differences in any outcomes of interest between the NSAID group and the unperturbed controls. ConclusionsNSAIDs negatively impact PFM regeneration in the preclinical SBI model. The above appears to be driven by the negative impact of NSAIDs on pelvic muscle stem cell function, resultant in delayed temporal progression of PFM regeneration following birth injury. These novel findings provide impetus to investigate the impact of postpartum NSAID administration on muscle regeneration in women at high risk for PFM injury.

The Impact of Anti-Inflammatory Drugs on the Prokaryotic Community Composition and Selected Bacterial Strains Based on Microcosm Experiments.

Non-steroidal anti-inflammatory drugs (NSAIDs) are increasingly recognized as potential environmental contaminants that may induce toxicity in aquatic ecosystems. This 3-week microcosm experiment explores the acute impacts of NSAIDs, including diclofenac (DCF), ibuprofen (IBU), and acetylsalicylic acid (ASA), on bacterial communities using a wide range of these substances (200-6000 ppm). The results showed that the NSAID-treated microcosms had higher cell count values than control samples, though the diversity of microbial communities decreased. The isolated heterotrophic bacteria mostly belonged to Proteobacteria, particularly Klebsiella. Next-generation sequencing (NGS) revealed that NSAIDs altered the structure of the bacterial community composition, with the proportion of Proteobacteria aligning with the selective cultivation results. Bacteria had higher resistance to IBU/ASA than to DCF. In DCF-treated microcosms, there has been a high reduction of the number of Bacteroidetes, whereas in the microcosms treated with IBU/ASA, they have remained abundant. The numbers of Patescibacteria and Actinobacteria have decreased across all NSAID-treated microcosms. Verrucomicrobia and Planctomycetes have tolerated all NSAIDs, even DCF. Cyanobacteria have also demonstrated tolerance to IBU/ASA treatment in the microcosms. The archaeal community structure was also impacted by the NSAID treatments, with Thaumarchaeota abundant in all microcosms, especially DCF-treated microcosms, while Nanoarchaeota is more typical of IBU/ASA-treated microcosms with lower NSAID concentrations. These results indicate that the presence of NSAIDs in aquatic environments could lead to changes in the composition of microbial communities.

Diclofenac Diminished the Unfolded Protein Response (UPR) Induced by Tunicamycin in Human Endothelial Cells.

Diclofenac belongs to the class of nonsteroidal anti-inflammatory drugs (NSAIDs), which are amongst the most frequently prescribed drugs to treat fever, pain and inflammation. Despite the presence of NSAIDs on the pharmaceutical market for several decades, epidemiological studies have shown new clinical applications of NSAIDs, and new mechanisms of their action were discovered. The unfolded protein response (UPR) activated under endoplasmic reticulum (ER) stress is involved in the pathophysiology of many diseases and may become a drug target, therefore, the study evaluated the effects of diclofenac on the tunicamycin-induced UPR pathways in endothelial cells. RT PCR analysis showed that diclofenac significantly inhibited activation of ER stress-responsive genes, i.e., CHOP/DITT3, GRP78/HSPA5 and DNAJB9. Additionally, the drug diminished the significant upregulation and release of the GRP78 protein, as evaluated using the ELISA assay, which was likely to be involved in the mechanism of the UPR activation resulting in apoptosis induction in endothelial cells. These results suggest the value of diclofenac as a factor capable of restoring the ER homeostasis in endothelial cells by diminishing the UPR.

Assessing the exposure to human and veterinary pharmaceuticals in waterbirds: The use of feathers for monitoring antidepressants and nonsteroidal anti-inflammatory drugs

Exposure to active pharmaceutical ingredients (APIs) from both human and veterinary sources is an increasing threat to wildlife welfare and conservation. Notwithstanding, tracking the exposure to pharmaceuticals in non-target and sensitive vertebrates, including birds, is seldom performed and relies almost exclusively on analysing internal organs retrieved from carcasses or from experimentally exposed and sacrificed birds. Clearly, this excludes the possibility of performing large-scale monitoring. Analysing feathers collected from healthy birds may permit this, by detecting APIs in wild birds, including protected and declining species of waterbirds, without affecting their welfare. To this end, we set up a non-destructive method for analysing the presence of non-steroidal anti-inflammatory drugs (NSAIDs), selective serotonin reuptake inhibitors (SSRIs) and noradrenaline reuptake inhibitors (SNRIs) in the feathers of fledglings of both the Mediterranean gull (Ichtyaetus melanocephalus) and the Sandwich tern (Thalasseus sandvicensis). The presence of several NSAIDs and SSRIs above the method quantification limits have confirmed that feathers might be a suitable means of evaluating the exposure of birds to APIs. Moreover, the concentrations indicated that waterbirds are exposed to NSAIDs, such as diclofenac, ibuprofen and naproxen, and SSRIs, such as citalopram, desmethylcitalopram, fluvoxamine and sertraline, possibly due to their widespread use and incomplete removal in wastewater treatment plants (WWTPs). The active ingredient diclofenac raises a the primary concern for the ecosystem and the welfare of the waterbirds, due to its high prevalence (100% and 83.3% in Mediterranean gull and Sandwich tern, respectively), its concentrations detected in feathers (11.9 ng g−1 and 6.7 ng g−1 in Mediterranean gull and Sandwich tern, respectively), and its documented toxicity toward certain birds.

Ionic covalent organic frameworks for Non-Steroidal Anti-Inflammatory drugs (NSAIDs) removal from aqueous Solution: Adsorption performance and mechanism

The presence of non-steroidal anti-inflammatory drugs (NSAIDs) is ubiquitous in ambient waterways due to their excessive consumption, which poses deleterious risks to human health and ecosystems. In this work, two classes of ionic covalent organic frameworks with different pore sizes (iCOFs) [TPA (terephthaldicarboxaldehyde)-TGCl (triaminoguanidinium chloride) and BPDA (4,4′-Biphenyldicarboxaldehyde)-TGCl], which is consisted of guanidine moieties as intrinsic cationic knots and different chain length dialdehydes, were innovatively exploited as adsorbents for the removal of NSAIDs. Studies revealed that the prepared iCOFs had a strong affinity for NSAIDs, particularly for diclofenac sodium (DCF) and ketoprofen (KT). The adsorption isotherms and kinetics data of both these iCOFs were in alignment with the Langmuir model and pseudo-second-order model, respectively. Guanidine-based iCOF knots trigger electrostatic interactions, which are the primarily intrinsic driving force that facilitates the efficient removal of DCF and KT. Furthermore, π-π stacking interactions are proposed based on iCOF skeletons. Additionally, TPA-TGCl was more preponderantly adsorptive toward DCF and KT, with maximum adsorption capacities of 724.64 mg·g−1 for DCF and 240.96 mg·g−1 for KT. Among them, the maximum adsorption capacity of DCF on TPA-TGCl was 27 times higher than that of carbon nanospheres, almost triple that of a magnetic cellulose ionomer/LDH, and two-fold higher than that of UiO-66-SO3H. Finally, the TPA-TGCl effectively achieved five adsorption–desorption cycles. Ultimately, iCOFs simultaneously meet the demands of structural regularity and accessibility to functional sites. Thus, they can be considered as highly efficient and recyclable adsorbents with potential applications for the remediation of the water environment, while providing a scientific basis for functional material design.

Plasma and serum oxylipin, endocannabinoid, bile acid, steroid, fatty acid and nonsteroidal anti-inflammatory drug quantification in a 96-well plate format

The goal of this research was to develop a high-throughput, cost-effective method for metabolic profiling of lipid mediators and hormones involved in the regulation of inflammation and energy metabolism, along with polyunsaturated fatty acids and common over-the-counter non-steroidal anti-inflammatory drugs (NSAIDs). We describe a 96-well plate protein precipitation and filtration procedure for 50 μL of plasma or serum in the presence of 37 deuterated analogs and 2 instrument internal standards. Data is acquired in two back-to-back UPLC-MS/MS analyses using electrospray ionization with positive/negative switching and scheduled multiple reaction monitoring for the determination of 145 compounds, including oxylipins, endocannabinoids and like compounds, bile acids, glucocorticoids, sex steroids, polyunsaturated fatty acids, and 3 NSAIDs. Intra- and inter-batch variability was <25% for >70% of metabolites above the LOQ in both matrices, but higher inter-batch variability was observed for serum oxylipins and some bile acids. Results for NIST Standard Reference Material 1950, compared favorably with the 20 certified metabolite values covered by this assay, and we provide new data for oxylipins, N-acylethanolamides, glucocorticoids, and 17-hydroxy-progesterone in this material. Application to two independent cohorts of elderly men and women showed the routine detection of 86 metabolites, identified fasting state influences on essential fatty acid-derived oxylipins, N-acylethanolamides and conjugated bile acids, identified rare presence of high and low testosterone levels and the presence of NSAIDs in ∼10% of these populations. The described method appears valuable for investigations in large cohort studies to provide insight into metabolic cross-talk between the array of mediators assessed here.

Pharmacology-informed prediction of the risk posed to fish by mixtures of non-steroidal anti-inflammatory drugs (NSAIDs) in the environment

The presence of non-steroidal anti-inflammatory drugs (NSAIDs) in the aquatic environment has raised concern that chronic exposure to these compounds may cause adverse effects in wild fish populations. This potential scenario has led some stakeholders to advocate a stricter regulation of NSAIDs, especially diclofenac. Considering their global clinical importance for the management of pain and inflammation, any regulation that may affect patient access to NSAIDs will have considerable implications for public health. The current environmental risk assessment of NSAIDs is driven by the results of a limited number of standard toxicity tests and does not take into account mechanistic and pharmacological considerations. Here we present a pharmacology-informed framework that enables the prediction of the risk posed to fish by 25 different NSAIDs and their dynamic mixtures. Using network pharmacology approaches, we demonstrated that these 25 NSAIDs display a significant mechanistic promiscuity that could enhance the risk of target-mediated mixture effects near environmentally relevant concentrations. Integrating NSAIDs pharmacokinetic and pharmacodynamic features, we provide highly specific predictions of the adverse phenotypes associated with exposure to NSAIDs, and we developed a visual multi-scale model to guide the interpretation of the toxicological relevance of any given set of NSAIDs exposure data. Our analysis demonstrated a non-negligible risk posed to fish by NSAID mixtures in situations of high drug use and low dilution of waste-water treatment plant effluents. We anticipate that this predictive framework will support the future regulatory environmental risk assessment of NSAIDs and increase the effectiveness of ecopharmacovigilance strategies. Moreover, it can facilitate the prediction of the toxicological risk posed by mixtures via the implementation of mechanistic considerations and could be readily extended to other classes of chemicals.

NSAIDs disrupt intestinal homeostasis by suppressing macroautophagy in intestinal epithelial cells

Small intestinal damage induced by nonsteroidal anti-inflammatory drugs (NSAIDs) remains an under-recognized clinical disorder. The incomplete understanding of the pathophysiology has hampered the development of prevention and treatment strategies leading to the high morbidity and mortality rates. NSAIDs are known to modulate macroautophagy, a process indispensable for intestinal homeostasis. Whether NSAIDs stimulate or repress macroautophagy and how this correlates with the clinical manifestations of NSAID enteropathy, however, remains unknown. The objectives of this study were to determine whether NSAIDs impaired macroautophagy and how this affects macroautophagy-regulated intestinal epithelial cell (IEC) processes essential for intestinal homeostasis (i.e., clearance of invading pathogens, secretion and composition of mucus building blocks, and inflammatory response). We show that NSAID treatment of IECs inhibits macroautophagy in vitro and in vivo. This inhibition was likely attributed to a reduction in the area and/or distribution of lysosomes available for degradation of macroautophagy-targeted cargo. Importantly, IEC regulatory processes necessary for intestinal homeostasis and dependent on macroautophagy were dysfunctional in the presence of NSAIDs. Since macroautophagy is essential for gastrointestinal health, NSAID-induced inhibition of macroautophagy might contribute to the severity of intestinal injury by compromising the integrity of the mucosal barrier, preventing the clearance of invading microbes, and exacerbating the inflammatory response.

Analysis of the assortment, socio-economic availability and volumes of drugs consumption of the nonsteroidal anti-inflammatory group on the Ukrainian pharmaceutical market

Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used in the world and in Ukraine. Anti-inflammatory and analgesic therapy of chronic musculoskeletal system diseases should be conducted for long periods of time. This requires the presence of NSAIDs that are characterized by distinct pharmacological properties and a high level of safety. Aim. To study the assortment, economic availability and volumes of NSAIDs consumption on the pharmaceutical market of Ukraine during the period of 2014-2018. Materials and methods. Data of the information and retrieval system “Morion” were used to analyze the assortment of NSAIDs. To analyze the availability of NSAIDs, the solvency adequacy ratio of Ca.s. was calculated, which means % of the average salary needed to pay for a monthly course of treatment with a certain drug. Consumption of drugs was carried out using the ATC / DDD methodology, which is a tool for determining the number of prescribed and taken for a specific time period daily doses of drugs (DDD) with a specific ATC code, which defines indications for use. Results were reported in standardized international units of DDDs/1000 inhabitants/day (DiD). Results. Studies have shown that NSAIDs on the pharmaceutical market are represented by 32 INN and 302-419 trade names (TNs). Most of the drugs are represented by foreign manufacturers. Ukrainian pharmaceutical companies presented 37 % of TN in 2018. The following INNs were presented during the study period by the largest number TN: Diclofenac (69-80), Ketorolac (14-18), Diclofenac Combinations (13-20), Meloxicam (52-66), Ibuprofen (58-66), Dexketoprofen (17-24), Nimesulide (21-25), Chondroitin sulfate (18-22). From 75 % to 86 % of TN on the market had high availability for the population in 2014-2018, from 3 to 4 % of TN – low availability. The volume of NSAID consumption use in Ukraine during 2014-2018 was significant: in 2014 – 16.67 DiD; in 2015 – 14.67 DiD; at 2016 – 16.37 DiD; in 2017 – 18.5 DiD; in 2018 – 20.4 DiD. The following drugs were most consumed in all five years of study: Diclofenac, Nimesulide, Ibuprofen, Ketorolac, Meloxicam. Among them, only Ibuprofen is a non-prescription drug and used in pediatrics as an antipyretic and analgesic drug. These are the INN that are on the Ukrainian market with the greatest number of TNs and that have advantages in terms of their pharmacological action and safety with long-term use for the treatment of chronic diseases. Conclusions. On the pharmaceutical market of Ukraine wide assortment of INN and TNs of NSAIDs are represented, consumed in large volumes, increasing every year. Among them, a large segment of TN based on five INN (Diclofenac, Nimesulide, Ibuprofen, Ketorolac and Meloxicam), which have marked clinical efficacy and are safe for long-term use, are the most consumed.

NSAID associated bilateral renal infarctions: a case report.

Renal infarctions (RIs) are caused by interruptions in the renal arterial blood flow. RIs are generally considered to be rare, however we present the case of a 37 year old woman whose renal infarction was likely due to the vasoconstrictive effects of non-steroidal anti-inflammatory drugs. Although high-dose non-steroidal anti-inflammatory drugs (NSAIDs) are known to cause a decrease in renal perfusion, they have not been accepted as causative agents in renal infarction. Theoretically, patients in prostaglandin dependent states should be more vulnerable to renovascular vasoconstriction and resulting hypoperfusion in the presence of NSAIDs. Given the high prevalence of NSAID use, we suspect that this mechanism of renal injury may be more prevalent than previously thought.

Bioremoval of non-steroidal anti-inflammatory drugs by Pseudoxanthomonas sp. DIN-3 isolated from biological activated carbon process

The presence of non-steroidal anti-inflammatory drugs (NSAIDs) in the environment is an emerging concern owing to their potential threat on aquatic ecosystems and living organisms. To investigate the bioremoval potential of a biological activated carbon (BAC) filter for the removal of NSAIDs, removal of diclofenac (DCF), ibuprofen (IBU), and naproxen (NAP) by biofilms on a bench-scaled BAC column operated for 400 days was studied. The results showed that the BAC column effectively removed the three NSAIDs (>90%). One bacterial strain isolated from the BAC, Pseudoxanthomonas sp. DIN-3, was able to simultaneously remove DCF, IBU, and NAP, which were supplied as the sole carbon source. In 14 days, 23%, 41%, and 39% of DCF, IBU, and NAP (50 μg L−1) were bioremoved, respectively, and strain DIN-3 eliminated IBU more rapidly than the other two NSAIDs. If only a single drug was added as the sole carbon source, ignoring the other drugs, the removal ability was overestimated by 5.0–27.0%. More efficient bioremoval was achieved, concomitantly with bacterial growth, via a co-metabolism with acetate, glucose, or methanol. Their intermediates were identified by UPLC-QQQ-MS, and their respective degradation pathways were also proposed. Moreover, based on the complete genome sequence of strain DIN-3, 49 related genes encoding the main enzymes involved in DCF, IBU, and NAP biodegradation were identified, including hemE, lpd, yihx, ligC, pobA, and ligA. These results suggested that Pseudoxanthomonas sp. DIN-3 is a potential degrader of DCF, IBU, and NAP, and to the best of our knowledge, this is the first report that demonstrates the bioremoval of DCF, IBU, and NAP simultaneously by an individual bacterial strain isolated from the environment. However, the bioremoval potential should be evaluated when assessing the applicability of the strain in the environment because of the combined effects of various pharmaceutical contaminants. The obtained results provide a foundation for the use of Pseudoxanthomonas sp. DIN-3 in the bioremoval of polycyclic NSAID-contaminated environments.

Ophthalmic Nonsteroidal Anti-Inflammatory Drugs as a Therapy for Corneal Dystrophies Caused by SLC4A11 Mutation.

SLC4A11 is a plasma membrane protein of corneal endothelial cells. Some mutations of the SLC4A11 gene result in SLC4A11 protein misfolding and failure to mature to the plasma membrane. This gives rise to some cases of Fuchs' endothelial corneal dystrophy (FECD) and congenital hereditary endothelial dystrophy (CHED). We screened ophthalmic nonsteroidal anti-inflammatory drugs (NSAIDs) for their ability to correct SLC4A11 folding defects. Five ophthalmic NSAIDs were tested for their therapeutic potential in some genetic corneal dystrophy patients. HEK293 cells expressing CHED and FECD-causing SLC4A11 mutants were grown on 96-well dishes in the absence or presence of NSAIDs. Ability of NSAIDs to correct mutant SLC4A11 cell-surface trafficking was assessed with a bioluminescence resonance energy transfer (BRET) assay and by confocal microscopy. The ability of mutant SLC4A11-expressing cells to mediate water flux (SLC4A11 mediates water flux across the corneal endothelial cell basolateral membrane as part of the endothelial water pump) was measured upon treatment with ophthalmic NSAIDs. BRET-assays revealed significant rescue of SLC4A11 mutants to the cell surface by 4 of 5 NSAIDs tested. The NSAIDs, diclofenac and nepafenac, were effective in moving endoplasmic reticulum-retained missense mutant SLC4A11 to the cell surface, as measured by confocal immunofluorescence. Among intracellular-retained SLC4A11 mutants, 20 of 30 had significant restoration of cell surface abundance upon treatment with diclofenac. Diclofenac restored mutant SLC4A11 water flux activity to the level of wild-type SLC4A11 in some cases. These results encourage testing diclofenac eye drops as a treatment for corneal dystrophy in patients whose disease is caused by some SLC4A11 missense mutations.

Toward Multifunctional Materials Incorporating Stepladder Manganese(III) Inverse-[9-MC-3]-Metallacrowns and Anti-Inflammatory Drugs.

The interaction of Mn(ClO4)2·6H2O with salicylaldoxime (H2sao) in the presence of nonsteroidal anti-inflammatory drug (NSAID) sodium diclofenac (Nadicl) or indomethacin (Hindo) leads to the formation of the hexanuclear Mn(III) clusters [Mn6(O)2(dicl)2(sao)6(CH3OH)6] (1) and [Mn6(O)2(indo)2(sao)6(H2O)4] (2) both characterized as stepladder inverse-9-metallacrown-3 accommodating dicl- or indo- ligands, respectively. When the interaction of MnCl2·4H2O with Nadicl or Hindo is in the absence of H2sao, the mononuclear Mn(II) complexes [Mn(dicl)2(CH3OH)4] (3) and [Mn(indo)2(CH3OH)4] (4) were isolated. The complexes were characterized by physicochemical and spectroscopic techniques, and the structure of complexes 1 and 2 was characterized by X-ray crystallography. Magnetic measurements (dc and ac) were carried out in order to investigate the nature of magnetic interactions between the magnetic ions and the overall magnetic behavior of the complexes.

Metabolic and Co-Metabolic Transformation of Diclofenac by Enterobacter hormaechei D15 Isolated from Activated Sludge.

The presence of non-steroidal anti-inflammatory drugs, such as diclofenac (DCF), in the environment, is an emerging problem due to their harmful effects on non-target organisms, even at low concentrations. We studied the biodegradation of DCF by the strain D15 of Enterobacter hormaechei. The strain was isolated from an activated sludge, and identified as E. hormaechei based on its physiological characteristics and its 16S RNA sequence. Using HPTLC and GC-MS methods, we demonstrated that this strain metabolized DCF at an elimination rate of 52.8%. In the presence of an external carbon source (glucose), the elimination rate increased to approximately 82%. GC-MS analysis detected and identified one metabolite as 1-(2,6-dichlorophenyl)-1,3-dihydro-2H-indol-2-one; it was produced as a consequence of dehydration and lactam formation reactions.

Nonsteroidal anti-inflammatory–induced inhibition of signal transducer and activator of transcription 6 (STAT-6) phosphorylation in aspirin-exacerbated respiratory disease

Aspirin desensitization provides long-term clinical benefits. The exact mechanisms of aspirin desensitization are not clearly understood. We sought to evaluate the effects of nonsteroidal anti-inflammatory drugs (NSAIDs) on T-cell activation of the IL-4 pathway in aspirin-sensitive patients with asthma and control subjects. A total of 11 aspirin-sensitive patients with asthma, 10 aspirin-tolerant patients with asthma, and 10 controls without asthma were studied. PBMCs were stimulated with an anti-CD3 antibody and IL-4 or IL-12, with and without the presence of NSAIDs. The expression of phosphorylated signal transducers and activators of transcription 6 (pSTAT6), phosphorylated signal transducers and activators of transcription 4, and IL-4 was detected in CD4 Tcells by flow cytometry. Stimulation with a combination of anti-CD3 and IL-4 induced pSTAT6 in CD4 Tcells from all subjects. The induction of pSTAT6 was significantly higher in aspirin-sensitive patients with asthma than in controls subjects. The increase in pSTAT6 was inhibited in a dose-dependent manner by aspirin and indomethacin and minimally by sodium salicylate. This inhibition was strongest in aspirin-sensitive patients. Two-group comparisons showed significant differences in pSTAT6 inhibition by all concentrations of indomethacin and aspirin: between aspirin-sensitive and aspirin-tolerant groups and between aspirin-sensitive and control groups. No differences were found between aspirin-tolerant and control groups at all 3 concentrations. The inhibition of pSTAT6 was associated with reduced IL-4 expression. NSAIDs inhibited signal transducers and activators of transcription 6 signaling in CD4 Tcells. This inhibition was significantly higher in aspirin-sensitive patients than in aspirin-tolerant subjects and was associated with reduced expression of IL-4. These findings have implications for clinical benefits of aspirin desensitization in aspirin-sensitive patients with asthma.

Safety of anti-tumor necrosis factor therapies in arthritis patients.

Inflammatory and rheumatic arthritis remain leading causes of disability worldwide. The arthritis therapeutic area commands the largest market for the prescription of biological and non-steroidal anti-inflammatory drugs (NSAID). Yet biotechnology and pharmaceutical companies conducting research and providing therapeutics in this area frequently face challenges in patient safety. The purpose of our study was to assess safety of anti-tumor necrosis factor therapies in arthritis patients. The present study systematically reviews adverse events of biologicals alone or in the presence of NSAIDs and other immunosuppressant therapeutics such as disease-modifying antirheumatic drugs (DMARD). We assessed the rheumatology literature that included clinical trials with anti-tumor necrosis factor (TNF) biologicals and case reports published between 2010 and 2014. Currently approved anti-TNF biologicals in arthritis include the monoclonal antibodies infliximab, adalimumab, certolizumab pegol and golimumab, and the fusion protein etanercept. The most frequently-reported adverse event was infection. We grouped the adverse reactions as immune-mediated, hypersensitivity syndrome reactions including cutaneous and hepatic manifestation, neurological, hematological, and malignancy. Most adverse events are due to the failure of host immunological control, which involves susceptibility to the drug itself, or de novo infection or reactivation of a latent bacterial or viral infection, often with a different expression of disease. Drug-induced liver injury associated with anti-TNF biologicals must be kept in mind when evaluating patients with increased liver enzymes. Risk assessment in individuals undergoing treatment with biologicals represents a step towards achieving a personalized medicine approach to identify those patients that will safely benefit from this therapeutic approach. Patients and physicians must be alert of anti-TNF agents as potential causes of drug-induced liver injury and monitor the therapies. Personalizing therapeutic pharmacovigilance promises to optimize benefits while minimizing side effects.

Bacterial degradation of naproxen – Undisclosed pollutant in the environment

The presence of non-steroidal anti-inflammatory drugs (NSAIDs) in the environment is an emerging problem due to their potential influence on human health and biocenosis. This is the first report on the biotransformation of naproxen, a polycyclic NSAID, by a bacterial strain. Stenotrophomonas maltophilia KB2 transformed naproxen within 35 days with about 28% degradation efficiency. Under cometabolic conditions with glucose or phenol as a carbon source degradation efficiency was 78% and 40%, respectively. Moreover, in the presence of naproxen phenol monooxygenase, naphthalene dioxygenase, hydroxyquinol 1,2-dioxygenase and gentisate 1,2-dioxygenase were induced. This suggests that degradation of naproxen occurs by its hydroxylation to 5,7,8-trihydroxynaproxen, an intermediate that can be cleaved by hydroxyquinol 1,2-dioxygenase. The cleavage product is probably further oxidatively cleaved by gentisate 1,2-dioxygenase. The obtained results provide the basis for the use of cometabolic systems in the bioremediation of polycyclic NSAID-contaminated environments.