ABSTRACT
Opiate alkaloids and their synthetic derivatives are still widely used in pain management, drug addiction, and abuse. To avoid serious side effects, compounds with properly designed pharmacological profiles at the opioid receptor subtypes are long needed. Here a series of 17-N-substituted derivatives of normorphine and noroxymorphone analogues with five- and six-membered ring substituents have been synthesized for structure-activity study. Some compounds showed nanomolar affinity to MOR, DOR and KOR in in vitro competition binding experiments with selective agonists [3H]DAMGO, [3H]Ile5,6-deltorphin II and [3H]HS665, respectively. Pharmacological characterization of the compounds in G-protein signaling was determined by [35S]GTPγS binding assays. The normorphine analogues showed higher affinity to KOR compared to MOR and DOR, while most of the noroxymorphone derivatives did not bind to KOR. The presence of 14-OH substituent resulted in a shift in the pharmacological profiles in the agonist > partial agonist > antagonist direction compared to the parent compounds. A molecular docking-based in silico method was also applied to estimate the pharmacological profile of the compounds. Docking energies and the patterns of the interacting receptor atoms, obtained with experimentally determined active and inactive states of MOR, were used to explain the observed pharmacological features of the compounds.
Subject(s)
Receptors, Opioid, mu , Receptors, Opioid , Receptors, Opioid, mu/metabolism , Molecular Docking Simulation , Receptors, Opioid/metabolism , Binding, Competitive , Structure-Activity Relationship , Receptors, Opioid, kappa/metabolismABSTRACT
There is still a need for synthetic approaches that are much faster, easier to scale up, more robust and efficient for generating gold(I)-thiolates that can be easily converted into gold-thiolate nanoclusters. Mechanochemical methods can offer significantly reduced reaction times, increased yields and straightforward recovery of the product, compared to the solution-based reactions. For the first time, a new simple, rapid and efficient mechanochemical redox method in a ball-mill was developed to produce the highly luminescent, pH-responsive Au(I)-glutathionate, [Au(SG)]n. The efficient productivity of the mechanochemical redox reaction afforded orange luminescent [Au(SG)]n in isolable amounts (mg scale), usually not achieved by more conventional methods in solution. Then, ultrasmall oligomeric Au10-12(SG)10-12 nanoclusters were prepared by pH-triggered dissociation of [Au(SG)]n. The pH-stimulated dissociation of the Au(I)-glutathionate complex provides a time-efficient synthesis of oligomeric Au10-12(SG)10-12 nanoclusters, it avoids high-temperature heating or the addition of harmful reducing agent (e.g., carbon monoxide). Therefore, we present herein a new and eco-friendly methodology to access oligomeric glutathione-based gold nanoclusters, already finding applications in biomedical field as efficient radiosensitizers in cancer radiotherapy.
ABSTRACT
A mechanochemical method is reported for the synthesis of Au(diphos)X complexes of diphosphine (diphos = XantPhos and N-XantPhos) ligands and halide ions (X = Cl and I). The Au(XantPhos)X (1: X = Cl; 2: X = I) and Au(N-XantPhos)Cl (3) complexes exhibited either yellowish green (1) or bluish green (2) emission, whereas 3 was seemingly non-emissive in the solid state at room temperature. Blue- (2B) and bluish green (2G) luminescent concomitant solvates of 2 were obtained by recrystallization. Luminescent colour changes from blue (2B) or bluish green (2G) to yellow were observed when these forms were subjected to mechanical stimulus, while the original emission colour can be recovered in the presence of solvent vapours. Moreover, the luminescence of 2B can be reversibly altered between blue and yellow by heating/cooling-cycles. These results demonstrate the power of mechanochemistry in the rapid (4 min reaction time), efficient (up to 98% yield) and greener synthesis of luminescent and stimuli-responsive gold(I) complexes.
ABSTRACT
Global DNA methylation and hydroxymethylation play an important role in gene expression. They can be connected with several diseases. The modification status could be a biomarker to determine the status of disease. A fast, easy and accurate liquid chromatography - tandem mass spectrometry method has been developed for the precise quantitation of 5-methylcytosine and 5-hydroxymethylcytosine. Formic acid was used for the hydrolysis of the DNA strand resulting in nucleobases. These polar hydrolysis products were separated on a normal phase column using reversed phase eluents in inverse gradient mode. Multiple reaction monitoring was applied to achieve high selectivity and sensitivity for the quantitation. A new relative quantitation model was developed by using guanine, as an internal standard, present in samples. The new method was successfully validated with excellent accuracy and precision values in the range of 0.005-0.5% for 5hmC and 1-15% for 5mC. The main advantages of this quantitation method are that, due to relative quantitation, calibration curves can be used without reacquiring the calibration points and no additional isotope labeled internal standards are required. The method was tested to identify the concentrations of 5mC and 5hmC in various sample types. The lowest level of DNA sample required in the case of 0.005% 5hmC is 0.5 µg.
Subject(s)
DNA Methylation , Guanine , Chromatography, Liquid , DNA , Tandem Mass SpectrometryABSTRACT
Sap beetle, Epuraea ocularis Fairmaire usually lays eggs and breeds on fermenting overripe fruits, and larvae pass through different instars before pupating on soil. In laboratory condition, mating pairs of adults copulated and females laid eggs in clusters; larva hatched out in 1 to 2 days, passed through four instars; mature larva migrated to soil for pupation. Larval development took about 12 to 17 days; and adult hatched out of pupa in about 4 to 5 days. Detailed morphology of egg, larva and pupa is presented herein, and significance of larva in taxonomy of beetles has been indicated.
Subject(s)
Coleoptera/classification , Animals , Coleoptera/growth & development , Female , India , Larva , Pupa , SoilABSTRACT
This article aims to outline the characteristics of the blood flow conveying copper (Cu) and gold (Au) nanoparticles (NPs) through a non-uniform endoscopic annulus with wall slip under the action of electromagnetic force and Hall currents. The flow of blood with the suspension of hybrid nanoparticles in the annulus is induced by the peristaltic pumping. The governing equations are modeled and then simplified with the postulate of lubrication theory. The resulting non-dimensional momentum equation after simplification is solved analytically by employing the He's homotopy perturbation method (HPM) with the computational software Mathematica program (version 11). The influential role of emerging physical parameters on the physiological features related to the blood flow is inferred graphically and physically. The analytical outcomes reveal that Hall parameter has a diminishing behavior on the blood flow while the inverse impact is endured for mounting Hartmann number. Electromagnetic field and Hall currents offer a superlative mode for regulating blood flow at the time of surgery. An increment in the volume fraction of nanoparticles causes a drop in the blood temperature profile. The trapping phenomenon is also explored with the help of contours. An expansion in Hartmann number reduces the size of entrapped bolus and ultimately vanishes when Hartmann number is very large. This prospective model may be applicable in electromagnetic micro-pumps, medical simulation devices, heart-lung machine (HLM), drug carrying and drug transport systems, cancer diagnosis, tumor selective photothermal therapy, etc.
Subject(s)
Blood Vessels/physiology , Copper/blood , Electromagnetic Fields , Endoscopy , Gold Alloys , Metal Nanoparticles , Models, Cardiovascular , Pulsatile Flow , Computer Simulation , Numerical Analysis, Computer-Assisted , Regional Blood Flow , Reproducibility of Results , Stress, Mechanical , TemperatureABSTRACT
The rheological perspective of blood flow with the suspension of metallic or non-metallic nanoparticles through arteries having cardiovascular diseases is getting more attention due to momentous applications in obstructed hemodynamics, nano-hemodynamics, nano-pharmacology, blood purification system, treatment of hemodynamic ailments, etc. Motivated by the novel significance and research in this direction, a mathematical hemodynamics model is developed to mimic the hemodynamic features of blood flow under the concentration of hybrid nanoparticles through an inclined artery with mild stenosis in the existence of dominating electromagnetic field force, Hall currents, heat source, and porous substance. Copper (Cu) and copper oxide (CuO) nanoparticles are submerged into the blood to form hybrid nano-blood suspension (Cu-CuO/blood). The attribute of the medium porosity on the blood flow is featured by Darcy's law. The mathematical equations describing the flow are formulated and simplified under mild stenosis and small Reynolds number assumptions. To determine the analytical solution of the resulting nonlinear momentum equation, the homotopy perturbation method (HPM) is employed. Several figures are graphed to assess the hemodynamical contributions of various intricate physical parameters on blood flow phenomena through the inclined stenosed artery. Significant outcomes from graphical elucidation envisage that the hemodynamic resistance to the blood flow is reduced due to the dispersion of more hybrid nanoparticles in the blood. The hemodynamic resistance (impedance) increases approximately two times by dispersing 0.11% hybrid nanoparticles in the blood flow. The temperature of Cu-CuO/blood is found to be lower in comparison to Cu-blood and pure blood. Intensification of Hall parameter attenuates the wall shear stress at the arterial wall. The trapping phenomena are also outlined via streamline plots which exemplify the blood flow pattern in the stenosed artery under the variation of the emerging parameters. As anticipated, the addition of a large number of hybrid nanoparticles significantly modulates the blood flow pattern in the stenotic region. The novel feature of this model is the impressive impact of Hall currents on hybrid nanoparticle doped blood flow through the stenosed artery. There is another piece of significance is that HPM is the most suitable method to handle the nonlinear momentum equation under the aforementioned flow constraints. Outcomes of this simulation may be valuable for advanced study and research in biomedical engineering, bio-nanofluid mechanics, nano-pharmacodynamics.
Subject(s)
Arterial Occlusive Diseases/physiopathology , Arteries/physiopathology , Copper/chemistry , Hemodynamics , Metal Nanoparticles , Models, Cardiovascular , Nanotechnology , Animals , Constriction, Pathologic , Humans , Porosity , Regional Blood Flow , Stress, MechanicalABSTRACT
Accumulating evidence indicate that astrocytes are essential players of the excitatory and inhibitory signaling during normal and epileptiform activity via uptake and release of gliotransmitters, ions, and other substances. Polyamines can be regarded as gliotransmitters since they are almost exclusively stored in astrocytes and can be released by various mechanisms. The polyamine putrescine (PUT) is utilized to synthesize GABA, which can also be released from astrocytes and provide tonic inhibition on neurons. The polyamine spermine (SPM), synthesized form PUT through spermidine (SPD), is known to unblock astrocytic Cx43 gap junction channels and therefore facilitate astrocytic synchronization. In addition, SPM released from astrocytes may also modulate neuronal NMDA, AMPA, and kainate receptors. As a consequence, astrocytic polyamines possess the capability to significantly modulate epileptiform activity. In this study, we investigated different steps in polyamine metabolism and coupled GABA release to assess their potential to control seizure generation and maintenance in two different epilepsy models: the low-[Mg2+] model of temporal lobe epilepsy in vitro and in the WAG/Rij rat model of absence epilepsy in vivo. We show that SPM is a gliotransmitter that is released from astrocytes and significantly contributes to network excitation. Importantly, we found that inhibition of SPD synthesis completely prevented seizure generation in WAG/Rij rats. We hypothesize that this antiepileptic effect is attributed to the subsequent enhancement of PUT to GABA conversion in astrocytes, leading to GABA release through GAT-2/3 transporters. This interpretation is supported by the observation that antiepileptic potential of the Food and Drug Administration (FDA)-approved drug levetiracetam can be diminished by specifically blocking astrocytic GAT-2/3 with SNAP-5114, suggesting that levetiracetam exerts its effect by increasing surface expression of GAT-2/3. Our findings conclusively suggest that the major pathway through which astrocytic polyamines contribute to epileptiform activity is the production of GABA. Modulation of astrocytic polyamine levels, therefore, may serve for a more effective antiepileptic drug development in the future.
ABSTRACT
Morphine and its derivatives play inevitably important role in the µ-opioid receptor (MOR) targeted antinociception. A structure-activity relationship study is presented for novel and known orvinol and thevinol derivatives with varying 3-O, 6-O, 17-N and 20-alkyl substitutions starting from agonists, antagonists and partial agonists. In vitro competition binding experiments with [3H]DAMGO showed low subnanomolar affinity to MOR. Generally, 6-O-demethylation increased the affinity toward MOR and decreased the efficacy changing the pharmacological profile in some cases. In vivo tests in osteoarthritis inflammation model showed significant antiallodynic effects of thevinol derivatives while orvinol derivatives did not. The pharmacological character was modelled by computational docking to both active and inactive state models of MOR. Docking energy difference for the two states separates agonists and antagonists well while partial agonists overlapped with them. An interaction pattern of the ligands, involving the interacting receptor atoms, showed more efficient separation of the pharmacological profiles. In rats, thevinol derivatives showed antiallodynic effect in vivo. The orvinol derivatives, except for 6-O-desmethyl-dihydroetorfin (2c), did not show antiallodynic effect.
Subject(s)
Analgesics, Opioid/pharmacology , Inflammation/drug therapy , Morphinans/pharmacology , Osteoarthritis/drug therapy , Receptors, Opioid, mu/agonists , Analgesics, Opioid/administration & dosage , Analgesics, Opioid/chemistry , Animals , Behavior, Animal/drug effects , Disease Models, Animal , Dose-Response Relationship, Drug , Female , Guinea Pigs , Male , Molecular Docking Simulation , Molecular Structure , Morphinans/administration & dosage , Morphinans/chemistry , Rats, Wistar , Receptors, Opioid, mu/antagonists & inhibitors , Structure-Activity RelationshipABSTRACT
Clinical evidence shows that following initial response to treatment, drug-resistant cancer cells frequently evolve and, eventually, most tumors become resistant to all available therapies. We compiled a focused library consisting of >500 commercially available or newly synthetized 8-hydroxyquinoline (8OHQ) derivatives whose toxicity is paradoxically increased rather than decreased by the activity of P-glycoprotein (Pgp), a transporter conferring multidrug resistance (MDR). Here, we deciphered the mechanism of action of NSC297366 that shows exceptionally strong Pgp-potentiated toxicity. Treatment of cells with NSC297366 resulted in changes associated with the activity of potent anticancer iron chelators. Strikingly, iron depletion was more pronounced in MDR cells due to the Pgp-mediated efflux of NSC297366-iron complexes. Our results indicate that iron homeostasis can be targeted by MDR-selective compounds for the selective elimination of multidrug resistant cancer cells, setting the stage for a therapeutic approach to fight transporter-mediated drug resistance. SIGNIFICANCE: Modulation of the MDR phenotype has the potential to increase the efficacy of anticancer therapies. These findings show that the MDR transporter is a "double-edged sword" that can be turned against resistant cancer.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/pharmacology , Iron Chelating Agents/pharmacology , Iron/metabolism , Neoplasms/drug therapy , Oxyquinoline/pharmacology , ATP Binding Cassette Transporter, Subfamily B/metabolism , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Cell Line, Tumor , Drug Resistance, Multiple/drug effects , Drug Resistance, Neoplasm/drug effects , Humans , Inhibitory Concentration 50 , Iron Chelating Agents/therapeutic use , Neoplasms/pathology , Oxyquinoline/analogs & derivatives , Oxyquinoline/therapeutic useABSTRACT
Introduction: "Aerotoxic syndrome" is a debated entity. Regulatory authorities consider long-term health effects to be an unlikely consequence of exposure to contaminated air because several air quality monitoring studies report low concentrations of toxic chemicals in cabin air. We describe two pilots and one flight attendant, who developed ill health during their flying career which improved after cessation of flying.Case details: The most frequently reported symptoms were headache, balance problems, fatigue, gastro-intestinal complaints and cognitive impairment. One of these patients had reduced levels of butyrylcholinesterase after a flight suggesting exposure to organophosphate compounds had occurred. All three were found to have elevated neuronal and glial auto-antibodies, biomarkers of central nervous system injury, and all three had genetic polymorphisms of paraoxonase (PON-1) and two of cytochrome P450, leading to a reduced ability to metabolize organophosphate compound (OPs).Discussion: A similar constellation of symptoms has been described in other studies of aircrew, although objective evidence of exposure is lacking in most of these studies. Reduced levels of butyrylcholinesterases in one of our cases is suggestive of causation and elevated neuronal and glial autoantibodies provide objective evidence of damage to the central nervous system. We consider further research is warranted.
Subject(s)
Air Pollutants, Occupational/adverse effects , Air Pollution, Indoor/adverse effects , Neurotoxicity Syndromes/etiology , Occupational Diseases/etiology , Occupational Exposure/adverse effects , Adult , Air Pollutants, Occupational/analysis , Air Pollution, Indoor/analysis , Aircraft , Female , Humans , Male , Middle Aged , Neurotoxicity Syndromes/diagnosis , Occupational Diseases/diagnosis , Occupational Exposure/analysis , PilotsABSTRACT
The use and significance of baicalin, the main bioactive component found in Radix Scutellaria, have been on the rise due to its interesting pharmacological properties. Baicalin, a low passive permeability compound, is directly absorbed from the upper intestine and its hepatic elimination is dominant. However, interaction but no transport studies have implicated organic aniontransporting polypeptides in its cellular uptake. By using mammalian cells stably expressing the uptake transporters of interest, we are showing that baicalin is a potent substrate of Organic aniontransporting polypeptide 2B1 (OATP2B1) and less potent substrate of OATP1B3. OATP2B1 and OATP1B3 transport baicalin and may play a role in the hepatic uptake of baicalin formed in the intestine.
Subject(s)
Flavonoids/metabolism , Organic Anion Transporters/metabolism , Solute Carrier Organic Anion Transporter Family Member 1B3/metabolism , Animals , Biological Transport , Dogs , HEK293 Cells , Humans , Intestinal Mucosa/metabolism , Liver/metabolism , Madin Darby Canine Kidney CellsABSTRACT
The aim of this study was to develop a sensitive, reliable and high-throughput liquid chromatography - electrospray ionization - mass spectrometric (LC-ESI-MS/MS) method for the simultaneous quantitation of cortisol and cortisone in human saliva. Derivatization with 2-hydrazino-1-methylpyridine (HMP) was one of the most challenging aspects of the method development. The reagent was reacting with cortisol and cortisone at 60°C within 1h, giving mono- and bis-hydrazone derivatives. Investigation of derivatization reaction and sample preparation was detailed and discussed. Improvement of method sensitivity was achieved with charged derivatization and use of on-line solid phase extraction (on-line SPE). The lower limit of quantitation (LLOQ) was 5 and 10pg/ml for cortisol and cortisone, respectively. The developed method was subsequently applied to clinical laboratory measurement of cortisol and cortisone in human saliva.
Subject(s)
Solid Phase Extraction , Chromatography, Liquid , Cortisone , Humans , Hydrocortisone , Saliva , Spectrometry, Mass, Electrospray Ionization , Tandem Mass SpectrometryABSTRACT
Localized gingival overgrowths are commonly encountered in our day-to-day clinical practice and often present a diagnostic dilemma to the clinicians. These lesions vary depending on the location, site, extent, histology, and/or etiopathology. Although most of the localized gingival enlargements represent the reactive lesion to plaque accumulation, the differential diagnosis ranges from peripheral fibroma to pyogenic granuloma to peripheral fibroma with ossification and/or calcification, peripheral giant cell granuloma, etc., Even the peripheral ameloblastoma may present clinically as a mere localized gingival enlargement. Therefore, proper histopathological diagnosis along with biopsy is necessary to effectively manage these lesions and to reduce their propensity for recurrence.
ABSTRACT
Herein, we give the very first example for the development of a fluorogenic molecular probe that combines the two-point binding specificity of biarsenical-based dyes with the robustness of bioorthogonal click-chemistry. This proof-of-principle study reports on the synthesis and fluorogenic characterization of a new, double-quenched, bis-azide fluorogenic probe suitable for bioorthogonal two-point tagging of small peptide tags by double strain-promoted azide-alkyne cycloaddition. The presented probe exhibits remarkable increase in fluorescence intensity when reacted with bis-cyclooctynylated peptide sequences, which could also serve as possible self-labeling small peptide tag motifs.
Subject(s)
Azides/chemistry , Fluorescent Dyes/chemistry , Oligopeptides/chemistry , Catalysis , Click Chemistry , Oligopeptides/metabolism , Staining and LabelingABSTRACT
Three species of Megauchenia Macleay [M. angustata (Erichson, 1843), M. quadricollis (Reitter, 1883) and M. indica (Grouvelle, 1908)] were studied based on a collection from India. The genus and the species are re-described. A key to the species of Megauchenia from India is appended.
Subject(s)
Coleoptera/anatomy & histology , Coleoptera/classification , Animals , Female , India , Male , Species SpecificityABSTRACT
Baicalein, the aglycone formed by hydrolysis of baicalin in the intestine, is well absorbed by passive diffusion but subjected to extensive intestinal glucuronidation. Efflux of baicalin, the low passive permeability glucuronide of baicalein from enterocytes, likely depends on a carrier-mediated transport. The present study was designed to explore potential drug-herb interaction by investigating the inhibitory effect of baicalin on the transport of reporter substrates by transporters and to identify the transporters responsible for the efflux of baicalin from enterocytes and hepatocytes. The interaction of baicalin with specific ABC transporters was studied using membranes from cells overexpressing human BCRP, MDR1, MRP2, MRP3 and MRP4. Baicalin was tested for its potential to inhibit vesicular transport by these transporters. The transport of baicalin by the selected transporters was also investigated. Transport by BCRP, MRP3 and MRP4 was inhibited by baicalin with an IC50 of 3.41 ± 1.83 µM, 14.01 ± 2.51 µM and 14.39 ± 5.69 µM respectively. Inhibition of MDR1 (IC50 = 94.84 ± 31.10 µM) and MRP2 (IC50 = 210.13 ± 110.49 µM) was less potent. MRP2 and BCRP are the apical transporters of baicalin that may mediate luminal efflux in enterocytes and biliary efflux in hepatocytes. The basolateral efflux of baicalin is likely mediated by MRP3 and MRP4 both in enterocytes and hepatocytes. Via inhibition of transport by ABC transporters, baicalin could interfere with the absorption and disposition of drugs.
Subject(s)
ATP-Binding Cassette Transporters/metabolism , Enterocytes/drug effects , Flavonoids/pharmacology , Hepatocytes/drug effects , Herb-Drug Interactions , Biological Transport/drug effects , Enterocytes/metabolism , Glucuronides/pharmacology , Hepatocytes/metabolism , HumansABSTRACT
The original aim of this study was to develop a method for the determination of baicalin from membrane vesicles. The unconventional chromatographic separation ("inverse gradient elution" on a reversed phase column) was due to a lucky chance, which is detailed and discussed in this study. The validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is proved to be sensitive, rapid and selective. Chromatographic separation was performed on a Zorbax SB-C8 column (250 mm × 4.6 mm, i.d.; 5 µm) with 0.1% formic acid in water and methanol by linear gradient elution. Quantification of baicalin was determined by multiple reaction monitoring (MRM) mode using electrospray ionization (ESI). The calibration curve was linear (r = 0.9987) over the concentration range from 1 to 1000 nM. The coefficient of variation and relative error of baicalin for intra- and inter-assay at three quality control (QC) levels were 2.0-10.2% and -6.1 to 6.7%, respectively. The lower limit of quantification (LLOQ) for baicalin was 1 nM (0.446 ng/ml), without preconcentration of the sample. This method was subsequently applied to vesicular transport assays of baicalin in membrane vesicles successfully. The developed method can open up new area of research in the chromatographic separation of flavonoids and their glucuronides.
Subject(s)
Drugs, Chinese Herbal/chemistry , Flavonoids/chemistry , Calibration , Chromatography, Liquid/methods , Hydrophobic and Hydrophilic Interactions , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization/methods , Tandem Mass Spectrometry/methodsABSTRACT
The study characterizes a collection of 67 neonatal septicaemic Escherichia coli isolates on the basis of phylogroup, serotype, virulence, antibiotic resistance and also the association of CTX-M-producing E. coli and the ST131 clone in a developing country. Phylogroups B2 and D were predominant (33% and 19%, respectively). The most prevalent virulence factors (VFs) were traT (69%) and iucC (68%) and most VFs were concentrated in the B2 isolates. High levels of resistance (⩾70%) to cefotaxime, ciprofloxacin and trimethoprim/sulfamethoxazole was recorded but meropenem remained the most active antimicrobial. Six (9%) of the study isolates belonged to the ST131 clone, five of which were from the same hospital, and were either indistinguishable or closely related by pulsed-field gel electrophoresis. Although the prevalence of CTX-M-15-producing isolates was high (81%), the ST131 clone was relatively infrequent (11%) in extended spectrum ß-lactamase (ESBL)-producers. The ST131 clone was characterized by the presence of bla CTX-M-15, qnrS, aac(6')-Ib-cr, IncF plasmids and virulence determinants such as iucC, papC, traT, usp, hlyA, iroN E.coli , cnf, and sat. We conclude that clonal spread of ST131 did not contribute directly to the high prevalence of CTX-M-15 in our settings.