Search Results (12)

Atherosclerosis (AS) is an important causative agent of cardiovascular diseases, and the occurrence and development of AS is accompanied by oxidative stress, so antioxidant therapy has become one of the strategies for the treatment of AS. This study aimed to design and construct an apolipoprotein ApoA1-modified inorganic–organic composite nanosystem for AS therapy, in which ApoA1 was modified onto carboxylated CeO₂/Mn₃O₄ by covalent bonding, resulting in an inorganic–organic nanocomplex with a structure similar to that of high-density lipoprotein. The nanocomplex could effectively deliver the antioxidant nanoparticles to the AS plaque through the specific recognition between ApoA1 and the macrophage at the AS lesion site. For one thing, the nanocomplex could alleviate the oxidative stress environment of the AS site through the highly efficient antioxidant effect of CeO₂/Mn₃O₄, which played a therapeutic role in the treatment of AS. For another, it could effectively eliminate the formed lipid plaques and maximally alleviate and treat AS by utilizing the cholesterol efflux effect of ApoA1. Full article

Background/Objectives: Previously, we reported that 3-O-alkyl difluoroquercetins (di-F-Q) potentiates the antimicrobial activity of aztreonam (ATM) against metallo-β-lactamase (MBL)-producing P. aeruginosa through simultaneous inhibition of MBLs and efflux pumps. However, the ATM-potentiating activity of the 3-O-alkyl di-F-Q was observed only at high and potentially toxic concentrations (32 mg/L). Methods: As both MBLs and efflux pumps reside in the periplasm of Gram-negative bacteria, their inhibitors should accumulate in the periplasmic space. However, the outer membrane porins, the major entry pathway in Gram-negative bacteria, allow the passive diffusion of hydrophilic polar molecules across the outer membrane. Thus, we reasoned that the introduction of a polar substituent at 7-OH position of 3-O-alkyl di-F-Q would enhance its periplasmic concentration to result in potentiation of ATM at lower concentrations. Results: The title compound 5 exhibited inhibitory activity against NDM-1 as well as the efflux pump of P. aeruginosa, which resulted in synergistical potentiation of ATM. A combination of ATM (8 mg/L) and 5 (8 mg/L) inhibited 80% of the ATM-resistant CPPA, while ATM alone did not show any inhibition. In addition, only 4 mg/L of 5 was needed to reduce the MIC₉₀ of ATM four-fold in ATM-resistant CPPA (n = 15). The time–kill data further supported the effectiveness of the combined treatment of ATM with 5, and the combination of ATM (1xMIC) with 8 mg/L of 5 showed bactericidal effects in every bacterial strain tested (PA-002, bla_IMP, PA-003, bla_VIM, PA-014, bla_GES, and PA-017, bla_NDM) by reducing the bacterial loads by 5.1 log₁₀~8.9 log₁₀. Conclusions: The title compound 5 exhibited inhibitory activity against NDM-1 as well as the efflux pump of P. aeruginosa, and the combined inhibitory activity resulted in synergistical potentiation of ATM. It should be noted that most CPPA isolates tested were sensitized to 8 mg/L of ATM upon combination with 4~8 mg/L of 5. Full article

Understanding carboxylate transport through lipid membranes under physiological conditions is critical in biomedicine and biotechnology, as it allows for the emulation of biological membrane functions and can enhance the absorption of hydrophobic carboxylate-based drugs. However, the structural diversity of carboxylates has made it challenging to study their transport, and the limited available examples do not provide a comprehensive understanding of the role of the organic moiety in this process. Here, we present an in-depth analysis of the diffusion and transport of various aliphatic and aromatic carboxylates into liposomes. We assessed the influence of their size, number of carboxylate groups, and presence of hydroxyl groups. Our findings from fluorescence assays, using lucigenin and HPTS as probes, revealed that most carboxylates can spontaneously diffuse into liposomes in their protonated state, facilitated by the efflux of HNO₃ when using NaNO₃ solutions at pH 7. The Cl-ISE assay showed chloride/carboxylate exchange by a synthetic anion transporter. Clear trends were observed when the organic moiety was systematically varied, with a particular enhancement of anion transport by the presence of hydroxyl groups in the aromatic carboxylates. Our findings provide insights into the processes by which carboxylates can enter liposomes, which can contribute to understanding the transport of other biologically relevant organic anions. Full article

Previous studies have shown that some lamellarin-resembling annelated azaheterocyclic carbaldehydes and related imino adducts, sharing the 1-phenyl-5,6-dihydropyrrolo[2,1-a]isoquinoline (1-Ph-DHPIQ) scaffold, are cytotoxic in some tumor cells and may reverse multidrug resistance (MDR) mediated by P-glycoprotein (P-gp). Herein, several novel substituted 1-Ph-DHPIQ derivatives were synthesized which carry carboxylate groups (COOH, COOEt), nitrile (CN) and Mannich bases (namely, morpholinomethyl derivatives) in the C2 position, as replacements of the already reported aldehyde group. They were evaluated for antiproliferative activity in four tumor cell lines (RD, HCT116, HeLa, A549) and for the ability of selectively inhibiting P-gp-mediated MDR. Lipophilicity descriptors and molecular docking calculations helped us in rationalizing the structure–activity relationships in the P-gp inhibition potency of the investigated 1-Ph-DHPIQs. As a main outcome, a morpholinomethyl Mannich base (8c) was disclosed which proved to be cytotoxic to all the tested tumor cell lines in the low micromolar range (IC₅₀ < 20 μM) and to inhibit in vitro the efflux pumps P-gp and MRP1 responsible for MDR, with IC₅₀s of 0.45 and 12.1 μM, respectively. Full article

The use of natural products isolated from mushrooms against infection, cancer diseases and other oxidative-stress-related diseases is one of the cornerstones of modern medicine. Therefore, we tried to establish a combination of medicinal mushrooms and nanotechnology possibly with the field of medicine for the development of antibacterial agents against these MDR strains. The aim of the research was to understand the molecular identification, characterization and antibacterial action of Calvatia gigantea and Mycena leaiana. The identification of fruiting body species via morpho-anatomical and molecular methods was necessary to analyze the genetic variability and phylogenetic relationships of mushrooms. Phylogenetic analysis revealed that Calvatia from Hunza, Pakistan, exhibited 98% resemblance to the previously discovered Langermannia gigantean (DQ112623) and L. gigantean (LN714562) from northern Europe, and Mycena (Pakistan) showed a 97% similarity to M. leaiana (MF686520) and M. leaiana (MW448623) from the USA. UV-vis, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) were used for AgNPs’ characterization. The UV-vis absorption peak of 500–600 nm indicates the AgNPs’ presence. XRD results determined Calvatia gigantea AgNPs were nanocrystals and Mycena leaiana seems to be amorphous. In addition, SEM results showed the cubic morphology of C. gigantea with a diameter of 65 nm, and the FTIR spectra of fruiting body revealed the presence of functional groups—carboxyl, nitro, and hydroxyl—in AgNPs, which catalyzed the reduction of Ag+ to Ag0. Further antibacterial activity of mushrooms against MDR strains was determined via agar well diffusion assay, and Minimum Inhibitory Concentration (MIC) was estimated by qualitative experimentation using the broth dilution method. All experiments were conducted in triplicate. The results showed that the mushroom AgNPs, along with their synergy and nano-composites (with the exception of Ethyl-acetate), were shown to have zones of inhibition from 4 mm to 29 mm against multidrug-resistant pathogens such as Acinetobacter baumannii, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus mirabilis, Enterobacter cloacae and Escherichia coli. The mushroom composites were active against most of the tested microorganisms whilst the lowest MIC value (10–40 mg/mL) was recorded against MDR strains. Hence, the present study suggested the possibility of employing compounds present in mushrooms for the development of new antibacterial agents, as well as efflux pump inhibitors. Full article

Multidrug resistance (MDR) is considered one of the major mechanisms responsible for the failure of numerous anticancer and antiviral chemotherapies. Various strategies to overcome the MDR phenomenon have been developed, and one of the most attractive research directions is focused on the inhibition of MDR transporters, membrane proteins that extrude cytotoxic drugs from living cells. Here, we report the results of our studies on a series newly synthesized of 5-arylidenerhodanines and their ability to inhibit the ABCB1 efflux pump in mouse T-lymphoma cancer cells. In the series, compounds possessing a triphenylamine moiety and the carboxyl group in their structure were of particular interest. These amphiphilic compounds showed over 17-fold stronger efflux pump inhibitory effects than verapamil. The cytotoxic and antiproliferative effects of target rhodanines on T-lymphoma cells were also investigated. A putative binding mode for 11, one of the most potent P-gp inhibitors tested here, was predicted by molecular docking studies and discussed with regard to the binding mode of verapamil. Full article

In this study, we investigated the delivery of synthetic neurosteroids into MCF-7 human breast adenocarcinoma cells via Organic Anionic Transporting Polypeptides (OATPs) (pH 7.4 and 5.5) to identify the structural components required for OATP-mediated cellular uptake and to get insight into brain drug delivery. Then, we identified structure-uptake relationships using in-house developed OATP1A2 homology model to predict binding sites and modes for the ligands. These binding modes were studied by molecular dynamics simulations to rationalize the experimental results. Our results show that carboxylic acid needs to be at least at 3 carbon-carbon bonds distance from amide bond at the C-3 position of the androstane skeleton and have an amino group to avoid efflux transport. Replacement of hydroxyl group at C-3 with any of the 3, 4, and 5-carbon chained terminal carboxylic groups improved the affinity. We attribute this to polar interactions between carboxylic acid and side-chains of Lys33 and Arg556. The additional amine group showed interactions with Glu172 and Glu200. Based on transporter capacities and efficacies, it could be speculated that the functionalization of acetyl group at the C-17 position of the steroidal skeleton might be explored further to enable OAT1A2-mediated delivery of neurosteroids into the cells and also across the blood-brain barrier. Full article

With the introduction of the new auxinic herbicide halauxifen-methyl into the oilseed rape (Brassica napus) market, there is a need to understand how this new molecule interacts with indigenous plant hormones (e.g., IAA) in terms of crop response. The aim of this study was to investigate the molecular background by using different growth conditions under which three different auxinic herbicides were administered. These were halauxifen-methyl (Hal), alone and together with aminopyralid (AP) as well as picloram (Pic). Three different hormone classes were determined, free and conjugated indole-3-acetic acid (IAA), aminocyclopropane carboxylic acid (ACC) as a precursor for ethylene, and abscisic acid (ABA) at two different temperatures and growth stages as well as over time (2–168 h after treatment). At 15 °C growth temperature, the effect was more pronounced than at 9 °C, and generally, the younger leaves independent of the developmental stage showed a larger effect on the alterations of hormones. IAA and ACC showed reproducible alterations after auxinic herbicide treatments over time, while ABA did not. Finally, a transcriptome analysis after treatment with two auxinic herbicides, Hal and Pic, showed different expression patterns. Hal treatment leads to the upregulation of auxin and hormone responses at 48 h and 96 h. Pic treatment induced the hormone/auxin response already after 2 h, and this continued for the other time points. The more detailed analysis of the auxin response in the datasets indicate a role for GH3 genes and genes encoding auxin efflux proteins. The upregulation of the GH3 genes correlates with the increase in conjugated IAA at the same time points and treatments. Also, genes for were found that confirm the upregulation of the ethylene pathway. Full article

The incessant pervasiveness of heavy metals in the environment is one of the precursory factors of pollution. This research study was endeavored upon to investigate the bioremediation potential of a nickel (Ni)-resistant bacterial isolate, identified as Bacillus altitudinis MT422188, whose optimum growth parameters were demonstrated at pH 7, temperature 32 °C, and 1 mM phosphate. Minimal Inhibitory Concentration (MIC) and EC₅₀ for Ni were observed to be 20 and 11.5 mM, respectively, whereas the cross heavy-metal resistance was discerned as Cu²⁺ (25 mM) > Zn²⁺ (15 mM) > Cr⁶⁺ (10 mM) > Pb²⁺ (5 mM) > Co²⁺ (8 mM) > Cd²⁺ (3 mM) > Hg²⁺ (0 mM). Ni biosorption studies by live and heat-killed bacterial cells were suggestive of Ni uptake being facilitated by an ATP-independent efflux system. A pilot-scale study displayed the effective removal of Ni (70 mg/L and 85 mg/L) at 4- and 8-day intervals, respectively. Moreover, chemotaxis and motility assays indicated the role of Ni as a chemoattractant for bacterial cells. The presence of Ni reduced the GR (0.001 ± 0.003 Ug⁻¹FW), POX (0.001 ± 0.001 Ug⁻¹FW), and SOD (0.091 ± 0.003 Ug⁻¹FW) activity, whereas Sodium dodecyl sulphate—Polyacrylamide gel electrophoresis (SDS-PAGE) revealed the presence of metallothionein (60 kDa). Kinetic and isotherm studies suggested a pseudo second-order and Freundlich model to be better fitted for our study. The thermodynamic parameters (∆H° = 3.0436 kJ/mol, ∆S° = 0.0224 kJ/mol/K) suggested the process to be endothermic, spontaneous, and favorable in nature. FTIR analysis elucidated the interaction of hydroxyl and carboxyl groups with Ni. Scanning Electron Microscopy (SEM) and Energy Dispersive X-Ray Spectroscopy (EDS) demonstrated changes in the morphological and elemental composition of the bacterial cells, which affirmed their interaction with Ni during biosorption. In summary, our study concludes the efficient role of Bacillus altitudinis MT422188 in removing Ni from polluted industrial effluents. Full article

Rhizosphere priming by terrestrial plants comprises increased or repressed efflux of CO₂ and N from soil organic matter (SOM), decaying under the impact of temperature, moisture, and the composition of rhizodeposits. Contemporarily, increases in water solubility vs. losses in molecular size, aromaticity, and the content in phenolic OH groups denote the degradation of SOM in planted soil. Root peroxidases (POs) and ‘polyphenoloxidases’ are surmised to contribute to these effects, however, final evidence for this is lacking. Therefore, seedlings of white mustard, alfalfa, and oilseed rape with wide spans in PO release were grown in hydroponic cultures at variable levels of Cu/Fe/Mn as Fenton metals, but also under P and Fe starvation to stimulate the release of carboxylic acids that form catalytic Mn³⁺ chelants from Mn²⁺ and MnO₂. The shortage in active oxygen as a cosubstrate of POs delayed the immediate oxidation of 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonate) (ABTS) supplements to the green ABTS^•+ by PO/H₂O₂, the possible formation of Mn³⁺ via PO catalyzed aryloxy radicals from root–released phenolics, and of HO^• by metal cations in H₂O₂ dependent Fenton–like reactions. Enhanced by exuded and external malate, O₂ independent MnO₂ supplements in some treatments formed ABTS^•+ spontaneously. The culture fluids then turned red in all treatments within 24–60 h by the formation of azodication (ABTS²⁺) derivatives in a second plant initiated oxidation step that is known to be catalyzed by substrate radicals. It is concluded that plants initiate oxidative activities that contribute to rhizosphere priming in an environment of oxidoreductase and carboxylate exudates, the indicated presence of mediating substrate radicals, and the cations and (hydr)oxides of transition metals. Pathways of H₂O₂ production upon the degradation of carboxylates and by the POs themselves are indicated. Full article

Carboxylate efflux from roots is a crucial and differential response of soybean genotypes to low phosphorus (P) stress. Exudation of carboxylic acids including oxalate, citrate, succinate and fumarate was induced under low P stress, particularly in P-efficient soybean genotypes. Enhancement of root length, surface area and volume further improved P acquisition under low P stress. To understand the molecular basis of carboxylate efflux under low P stress, the root proteome of contrasting genotypes (P-efficient: EC-232019 and P-inefficient: EC-113396) was compared. Among a total of 325 spots, 105 (32%) were differentially abundant proteins (DAPs) between sufficient (250 µM) and low P (4 µM) levels. Abundance of 44 (14%) proteins decreased by more than two-fold under low P stress, while 61 (19%) proteins increased by more than two-fold. Protein identification and annotation revealed that the DAPs were involved in a myriad of functions including carboxylic acid synthesis, carbohydrate, protein and lipid metabolism. Proteins with significant abundance included malate dehydrogenase, isocitrate dehydrogenase, phosphoglucomutase, phosphoglycerate mutase, fructokinase, enolase, phosphoglycerate kinase, triosephosphate isomerase, alcohol dehydrogenase, glucan water dikinase, glutamine synthetase and argininosuccinate lyase. Inferences from proteomic analysis suggests the crosstalk between various metabolic pathways implicated in conferring superior P acquisition efficiency under stress. Full article

The iron acquisition systems in Pseudomonas aeruginosa are inducible in response to low-iron conditions and important for growth of this organism under iron limitation. OprM is the essential outer membrane subunit of the MexAB-OprM xenobiotic efflux pump. We designed and constructed a new model antimicrobial screening system targeting both the iron-uptake system and xenobiotic efflux pumps. The oprM gene was placed immediately downstream of the ferri-pyoverdine receptor gene, fpvA, in the host lacking chromosomal oprM and the expression of oprM was monitored by an antibiotic susceptibility test under iron depleted and replete conditions. The recombinant cells showed wild-type susceptibility to pump substrate antibiotics, e.g., aztreonam, under iron limitation and became supersusceptible to them under iron repletion, suggesting that expression of oprM is under control of the iron acquisition system. Upon screening of a chemical library comprising 2952 compounds using this strain, a compound—ethyl 2-(1-acetylpiperidine-4-carboxamido)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylate—was found to enhance the efficacy of aztreonam under iron limitation, suggesting that the compound inhibits either the iron acquisition system or the MexAB-OprM efflux pump. This compound was subsequently found to inhibit the growth of wild-type cells in the presence of sublethal amounts of aztreonam, regardless of the presence or absence of dipyridyl, an iron-chelator. The compound was eventually identified to block the function of the MexAB-OprM efflux pump, showing the validity of this new method. Full article