Energy-dependent Mechanism Research Articles

Ultrasmall Carbon Nanodots as Theranostic Nanoheaters for Precision Breast Cancer Phototherapy: Establishing the Translational Potential in Tumor-in-a-Dish Models.

This study investigates the remarkable attributes of sulfur-doped carbon nanodots (CDs) synthesized in high yield and a narrow size distribution (4.8 nm). These CDs exhibit notable features, including potential bioelimination through renal clearance and efficient photothermal conversion in the near-infrared region with multicolor photoluminescence across the visible spectrum. Our research demonstrates high biocompatibility and effective near-infrared (NIR)-triggered photothermal toxicity when targeting mammospheres and patient-derived tumor organoids. Moreover, the study delves into the intricate cellular responses induced by CD-mediated hyperthermia. This involves efficient tumor mass death, activation of the p38-mitogen-activated protein kinase (MAPK) pathway, and upregulation of genes associated with apoptosis, hypoxia, and autophagy. The interaction of CDs with mammospheres reveals their ability to penetrate the complex microenvironment, impeded at 4 °C, indicating an energy-dependent endocytosis mechanism. This observation underscores the CDs' potential for targeted drug delivery, particularly in anticancer therapeutics. This investigation contributes to understanding the multifunctional properties of sulfur-doped CDs and highlights their promising applications in cancer therapeutics. Utilizing 3-D tumor-in-a-dish patients' organoids enhances translational potential, providing a clinically relevant platform for assessing therapeutic efficacy in a context mirroring the physiological conditions of cancerous tissues.

Solid self-microemulsifying drug delivery system (S-SMEDDS) prepared by spray drying to improve the oral bioavailability of cinnamaldehyde (CA)

The aim of this study was to prepare a solid self-microemulsifying drug delivery system (S-SMEDDS) of cinnamaldehyde (CA) by spray drying technique to improve the oral bioavailability of CA. The preparation of CA S-SMEDDS with maltodextrin as the solid carrier, a core-wall material mass ratio of 1:1, a solid content of 20% (w/v), an inlet air temperature of 150 °C, an injection speed of 5.2 mL/min, and an atomization pressure of 0.1 MPa was determined by using the encapsulation rate as the index of investigation. Differential scanning calorimetry (DSC) revealed the possibility of CA being encapsulated in S-SMEDDS in an amorphous form. The in-vitro release showed that the total amount of CA released by S-SMEDDS was approximately 1.3 times higher than that of the CA suspension. Pharmacokinetic results showed that the relative oral bioavailability of CA S-SMEDDS was also increased to 1.6-fold compared to CA suspension. Additionally, we explored the mechanism of CA uptake and transport of lipid-soluble drugs CA by S-SMEDDS in a Caco-2/HT29 cell co-culture system for the first time. The results showed that CA S-SMEDDS uptake on the co-culture model was mainly an energy-dependent endocytosis mechanism, including lattice protein-mediated endocytosis and vesicle-mediated endocytosis. Transport experiments showed that CA S-SMEDDS significantly increased the permeability of CA in this model. These findings suggested that CA S-SMEDDS is an effective oral solid dosage form for increasing the oral bioavailability of lipid-soluble drug CA.

In Vitro and In Vivo Antitumor Assay of Mitochondrially Targeted Fluorescent Half-Sandwich Iridium(III) Pyridine Complexes

Half-sandwich iridium(III) complexes show potential value in the anticancer field. However, complexes with favorable luminescence performance are rare, which limits further investigation of the anticancer mechanism. In this paper, 10 triphenylamine-modified fluorescent half-sandwich iridium(III) pyridine complexes {[(η5-Cpx)Ir(L)Cl2]} (Ir1-Ir10) were prepared and showed potential antiproliferative activity, effectively inhibiting the migration of A549 cells. Ir6, showing the best activity among these complexes, exhibited excellent fluorescence performance (absolute fluorescence quantum yield of 15.17%) in solution. Laser confocal detection showed that Ir6 followed an energy-dependent cellular uptake mechanism, specifically accumulating in mitochondria (Pearson co-localization coefficient of 0.95). A Western blot assay further confirmed the existence of a mitochondrial apoptotic channel. Additionally, Ir6 could arrest the cell cycle at the G2/M phase, catalyze NADH oxidation, reduce the mitochondrial membrane potential, induce an increase in the level of intracellular reactive oxygen species, and exhibit a mechanism of oxidation. An in vivo antitumor assay confirmed that Ir6 can effectively inhibit tumor growth and is safer than cisplatin.

P-Glycoprotein Activity at Diagnosis Does Not Predict Therapy Outcome and Survival in Canine B-Cell Lymphoma.

Simple SummaryClinical experience in human and canine clinics shows that following initial response to treatment, drug-resistant cancer cells frequently evolve and eventually, most tumors become resistant to all available therapies. The most straightforward cause of therapy resistance is linked to cellular alterations that prevent drugs from acting on their target. Drug efflux mediated by the ABC transporter P-glycoprotein (P-gp) contributes to unfavorable treatment outcome in several human malignancies. Here, we characterize a large cohort of canine B-cell lymphoma patients followed for over 7 years. We show that the intrinsic P-gp activity of tumor cells characterized at the time of diagnosis is not predictive for therapy outcome. Our results highlight the complexity of clinical drug resistance mechanisms and suggests that the relevance of P-gp in acquired resistance should be further investigated by the continuous monitoring of tumor cells during treatment.Various mechanisms are known to be involved in the development of multidrug resistance during cancer treatment. P-glycoprotein (P-gp) decreases the intracellular concentrations of cytotoxic drugs by an energy-dependent efflux mechanism. The aim of this study was to investigate the predictive value of P-gp function based on the evaluation of P-gp activity in tumor cells obtained from canine B-cell lymphoma patients at diagnosis. P-gp function of 79 immunophenotyped canine lymphoma samples was determined by flow cytometry using the Calcein assay. Dogs were treated with either the CHOP or the L-CHOP protocol, a subset of relapsed patients received L-asparaginase and lomustine rescue treatments. Among the 79 dogs, the median overall survival time was 417 days, and the median relapse-free period was 301 days. 47 percent of the samples showed high P-gp activity, which was significantly higher in Stage IV cancer patients compared to Stage II + III and V. Whereas staging was associated with major differences in survival times, we found that the intrinsic P-gp activity of tumor cells measured at diagnosis is not predictive for therapy outcome. Further studies are needed to identify the intrinsic and acquired resistant mechanisms that shape therapy response and survival in B-cell canine lymphoma patients.

Lysosomal Targeted Cyclometallic Iridium(Ⅲ) Salicylaldehyde-Coumarin Schiff Base Complexes and Anticancer Application.

Natural coumarin derivatives and cyclometallic iridium (Ⅲ) (IrⅢ) complexes have attracted much attention in the field of anticancer. In this study, six coumarin-modified cyclometallic IrⅢ salicylaldehyde Schiff base complexes ([(ppy)2Ir(O^N)]/[(ppy-CHO)2Ir(O^N)]) were designed and synthesized. Compared with coumarin and IrⅢ complex monomers, target complexes exhibited favorable cytotoxic activity toward A549 and BEAS-2B cells. These complexes could induce extensive apoptosis of A549 cell (late apoptosis), which was represented by the disturbance of cell cycle (G1-phase) and the accumulation of intracellular reactive oxygen species, exhibiting an anticancer mechanism of oxidation. With the help of suitable fluorescence of these complexes, no conflict with the probes, confocal detection confirmed that complexes showed an energy-dependent cellular uptake mechanism and triggered lysosome-mediated apoptosis in A549 cell line. Above all, our findings reveal the design of a lysosomal targeting cyclometallic IrⅢ Schiff base complexes and provide a new idea for the design of integrated drugs for diagnosis and treatment.

The thiosemicarbazone, DpC, broadly synergizes with multiple anti-cancer therapeutics and demonstrates temperature- and energy-dependent uptake by tumor cells

BackgroundThe di-2-pyridylketone thiosemicarbazones, di-2-pyridylketone-4,4-dimethyl-3-thiosemicarbazone (Dp44mT) and di-2-pyridylketone-4-cyclohexyl-4-methyl-3-thiosemicarbazone (DpC), demonstrate potent and selective anti-tumor activity. In fact, DpC entered Phase I clinical trials for advanced and resistant tumors. MethodsThis investigation examined the activity of these thiosemicarbazones in five tumor cell-types compared to nine clinically used chemotherapeutics and also in combination with these drugs. ResultsDp44mT and especially DpC demonstrated potent anti-proliferative activity that was significantly greater than a range of standard anti-cancer therapeutics. As most anti-cancer drugs are given in combination, further studies were performed to examine the synergistic activity of DpC or Dp44mT with these chemotherapeutics. Combination experiments revealed broad synergy between Dp44mT or DpC upon addition of these drugs, with a sequential protocol of treating first with standard chemotherapies followed by incubation with the thiosemicarbazones being optimal. However, combining DpC and Dp44mT resulted in a pronounced antagonistic drug interaction. To dissect the mechanism of this latter effect, custom-prepared 14C-DpC was implemented and examined for its uptake by cells. The avid uptake of 14C-DpC by tumor cells observed at 37 °C was suppressed at 4 °C and by the metabolic inhibitor, sodium fluoride, suggesting a temperature- and energy-dependent mechanism. Furthermore, competition studies using an excess of unlabeled Dp44mT or DpC inhibited 14C-DpC or 14C-Dp44mT uptake, respectively, suggesting these ligands utilize the same carrier/receptor, antagonizing the internalization of each other. Conclusions and general significanceThese studies demonstrate the potent and broad anti-proliferative activity of Dp44mT and particularly DpC, and are important for establishing optimized combinations with standard chemotherapies.

Extracellular Vesicles From Microalgae: Uptake Studies in Human Cells and Caenorhabditis elegans

Extracellular vesicles (EVs) are lipid membrane nano-sized vesicles secreted by various cell types for intercellular communication, found in all kingdoms of life. Nanoalgosomes are a subtype of EVs derived from microalgae with a sustainable biotechnological potential. To explore the uptake, distribution and persistence of nanoalgosomes in cells and living organisms, we separated them from a culture of the chlorophyte Tetraselmis chuii cells by tangential flow filtration (TFF), labelled them with different lipophilic dyes and characterized their biophysical attributes. Then we studied the cellular uptake of labelled nanoalgosomes in human cells and in C. elegans, demonstrating that they enter the cells through an energy dependent mechanism and are localized in the cytoplasm of specific cells, where they persist for days. Our data confirm that nanoalgosomes are actively uptaken in vitro by human cells and in vivo by C. elegans cells, supporting their exploitation as potential nanocarriers of bioactive compounds for theranostic applications.

Internalization of benzylisoquinoline alkaloids by resting and activated bone marrow-derived mast cells utilizes energy-dependent mechanisms

Objective and designDrug delivery to inflammatory cells is dependent upon poorly understood, complex endocytic processes. Berberine (BBR), a benzylisoquinoline alkaloid, binds to heparin and targets glycosaminoglycan-rich granules in mast cells (MC), but the mechanism of BBR internalization is unknown.MethodsBMMC were treated with various concentrations of BBR for different amounts of time and BBR internalization was assessed by flow cytometry and fluorescence microscopy. BMMC were pretreated with endocytic inhibitors or a growth factor (IL-3) prior to BBR exposure to access mechanisms of its internalization.ResultsAfter 24 h, 48 ± 0.8% of BMMC internalized BBR and this process was dependent upon temperature and the presence of glucose in the medium. Methanol fixation reduced BBR internalization, suggesting the involvement of an energy-dependent active transport mechanism. To determine mode of internalization, BBR was encapsulated into Lipofectamine TM lipoplexes since these are known to circumvent classical endocytic pathways. Incorporating BBR into lipoplexes decreased BBR internalization by 26% and 10% (10 μg/ml and 100 μg/ml Lipo-BBR respectively) by BMMC. BBR endocytosis was significantly reduced by Latrunculin B (88%), Cytochalasin B (87%), Chloroquine (86.5%) and 3-methyladenine (91%), indicating that actin polymerization, lysosomal pH and lysosomal self-degradation via the autophagy pathway was involved. In contrast, IL-3 treatment significantly enhanced BBR endocytosis (54% by 40 ng/ml IL-3) suggesting that IL-3 signaling pathways play a role in internalization.ConclusionsOur data suggests that internalization of BBR by resting and IL-3-activated BMMC utilizes an energy-dependent pathway that is dependent upon glucose metabolism and temperature. Furthermore, this process requires actin polymerization and lysosomal trafficking. These data suggest internalization of benzylisoquinoline compounds is an active and complex process.

Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli

Cell-penetrating peptides (CPPs) are increasingly used for cellular drug delivery in both pro- and eukaryotic cells, and oligoarginines have attracted special attention. How arginine-rich CPPs translocate across the cell envelope, particularly for prokaryotes, is still unknown. Arginine-rich CPPs efficiently deliver antimicrobial peptide nucleic acid (PNA) to its intracellular mRNA target in bacteria. We show that resistance to PNA conjugated to an arginine-rich CPP in Escherichia coli requires multiple genetic modifications and is specific for the CPP part and not to the PNA part. An integral part of the resistance was the constitutively activated Cpx-envelope stress response system (cpx∗), which decreased the cytoplasmic membrane potential. This indicates an indirect energy-dependent uptake mechanism for antimicrobials conjugated to arginine-rich CPPs. In agreement, cpx∗ mutants showed low-level resistance to aminoglycosides and an arginine-rich CPP conjugated to a peptide targeting the DNA sliding clamp, i.e., similar uptake in E. coli for these antimicrobial compounds.

Internalization of benzylisoquinoline alkaloids by resting and activated bone marrow-derived mast cells utilizes energy-dependent mechanisms

Abstract Berberine (BBR), a benzylisoquinoline alkaloid, binds to heparin and targets glycosaminoglycan-rich granules of myeloid-derived mast cells, but, the mechanism of berberine’s internalization is poorly understood. Our data showed that 84±3.3% of highly granulated bone marrow-derived mast cells (BMMC) internalized BBR (10 ug/mL) after 45 hr. Methanol fixation completely blocked this process, suggesting an energy-dependent active transport mechanism, and 3-mtheyladenine (600 uM) inhibited internalization by ~43%, indicating that lysosomal self-degradation via the autophagy pathway was involved. BBR incorporation into lipoplexes did not improve BBR internalization, but, rather decreased it by ~30% (1 ug/mL Lipo/BBR; n=3, p<0.05), suggesting that BBR internalization may utilize clathrin-dependent endosomal endocytosis. Interleukin-3 (IL-3)-mediated activation of BMMC augmented BBR internalization (~54% by 40 ng/mL IL-3) in a concentration-dependent manner, further suggesting that clathrin-mediated endocytosis as well as active cell division, have a profound role in BBR internalization. Altogether, our data suggest that internalization of BBR in resting and IL-3 activated BMMC, utilizes clathrin-dependent pathway. Our findings provide important new mechanistic insights into the internalization of benzylsoquinoline compounds by resting and activated mast cells and open new avenues for designing organelle-targeted nanotherapeutics.

LDL-conjugated to GM1 micelles incorporating anticancer drugs to improve tumor cell uptake

Objective(s): The role of lipoproteins (LDL) as active molecules with preferential tumor interaction, but limited drug delivery capacity, has been previously reported. On the other hand, in a previous report, we demonstrated the high capacity of monosialogangliosides (GM1) micelles as drug transporters. Materials and Methods: In this work, GM1 was loaded with high doses of oncologic drugs such Paclitaxel or Doxorubicin and binded to LDL lipoproteins to form GM1-drug-LDLwater soluble complex. Evidence suggests that both, hydrophobic and electrostatic forces, participate in the interaction, regulated by conditions such as pH, temperature and ionic strength.Results: Results of DLS and TEM show that GM1-LDL complexes are considerably larger than the sum of their individual compounds, with a high charge of electronegative surface (-55.9 mV). In addition, the cytotoxic effect on cell cultures is greater when drugs are contained in GM1-LDL complexes than when loaded in GM1 micelles. Conclusion: The results suggest the participation of active energy-dependent mechanism in the uptake of GM1-LDL drug, probably linked to the LDL receptor by the tumor cells. However, we could not confirm that the transport through LDL receptors is the only one that participates in the cellular uptake of the micelles.

Neonatal Rotenone Administration Induces Psychiatric Disorder-Like Behavior and Changes in Mitochondrial Biogenesis and Synaptic Proteins in Adulthood.

Since psychiatric disorders are associated with changes in the development of the nervous system, an energy-dependent mechanism, we investigated whether mitochondrial inhibition during the critical neurodevelopment window in rodents would be able to induce metabolic alterations culminating in psychiatric-like behavior. We treated male Wistar rat puppies (P) with rotenone (Rot), an inhibitor of mitochondrial complex I, from postnatal days 5 to 11 (P5-P11). We demonstrated that at P60 and P120, Rot-treated animals showed hyperlocomotion and deficits in social interaction and aversive contextual memory, features observed in animal models of schizophrenia, autism spectrum disorder, and attention deficit hyperactivity disorder. During adulthood, Rot-treated rodents also presented modifications in CBP and CREB levels in addition to a decrease in mitochondrial biogenesis and Nrf1 expression. Additionally, NFE2L2-activation was not altered in Rot-treated P60 and P120 animals; an upregulation of pNFE2L2/ NFE2L2 was only observed in P12 cortices. Curiously, ATP/ADP levels did not change in all ages evaluated. Rot administration in newborn rodents also promoted modification in Rest and Mecp2 expression, and in synaptic protein levels, named PSD-95, Synaptotagmin-1, and Synaptophysin in the adult rats. Altogether, our data indicate that behavioral abnormalities and changes in synaptic proteins in adulthood induced by neonatal Rot administration might be a result of adjustments in CREB pathways and alterations in mitochondrial biogenesis and Nrf1 expression, rather than a direct deficiency of energy supply, as previously speculated. Consequently, Rot-induced psychiatric-like behavior would be an outcome of alterations in neuronal paths due to mitochondrial deregulation.

Preparation and antitumor application of N-phenylcarbazole/triphenylamine-modified fluorescent half-sandwich iridium(III) Schiff base complexes.

Four N-phenylcarbazole/triphenylamine-appended half-sandwich iridium(III) salicylaldehyde Schiff base complexes ([(η5-Cpx)Ir(O^N)Cl]) were prepared and characterized. The complexes exhibited similar antitumor activity to cisplatin and effectively inhibited the migration of tumor cells. Furthermore, the complexes showed favourable hydrolytic activity, while remaining relatively stable in the plasma environment, which facilitated the binding of serum proteins and transport through them. These complexes could decrease the mitochondrial membrane potential, catalyze the oxidation of nicotinamide adenine dinucleotide, induce an increase in intracellular reactive oxygen species (ROS), and eventually result in apoptosis. Aided by their suitable fluorescence property, laser confocal detection showed that the complexes followed an energy-dependent mechanism for their cellular uptake, effectively accumulating in the lysosome and leading to lysosomal damage. In summary, the half-sandwich iridium(III) salicylaldehyde Schiff base complexes could induce lysosomal damage, increase intracellular ROS, and lead to apoptosis, which contributed to their antitumor mechanism of oxidation.

Non-pigmented ciliary epithelium derived extracellular vesicles uptake mechanism by the trabecular meshwork.

Consistent with increasing findings, extracellular vesicles (EVs), consider as a major constituents of the aqueous humor, have a role as signaling mediators in glaucoma. Following secretion, EVs hold immense promise for utilization as bio-therapeutics and drug delivery vehicles due to their nature as biological nanoparticles that facilitate intercellular molecular transport. Yet, the specific pathway utilizing for transferring signals by EVs in the ocular drainage system is not fully understood. Hence, the objective of this study was to examine internalization mechanisms by which Non-Pigmented Ciliary Epithelium (NPCE)-derived EVs deliver their signals to the Trabecular Meshwork (TM) cells. EVs were isolated and size and concentration were determined. Internalization study of treated EVs with Proteinase-K to achieve removal of surface membrane proteins on EVs was conducted. Energy dependent uptake mechanism was examined under various temperatures. Using uptake inhibitors endocytosis, phagocytosis, and Wnt-TGFβ2 signaling were investigated. TM cells exposed to NPCE EVs demonstrate a significant decrease in the levels of two proteins in two Wnt-TGFb2 signaling proteins levels: p-GSK3β and β-catenin. A significant decrease in the uptake by TM cells of Proteinase-K-treated EVs was found, followed by attenuation of the Wnt-TGFβ2 proteins expression. Energy dependent uptake revealed a significant decrease in EVs internalization. The exposure of TM cells to endocytosis uptake inhibitors abolished the decrease of the Wnt-TGFβ2 proteins levels. Exposure to phagocytosis uptake inhibitor resulted in a partial inhibition of NPCE EVs effect in TM cells. The attenuation of proteins expression levels following uptake inhibitors treatment or EVs membrane proteins removal indicates that Wnt-TGFβ2 signaling in TM cells is mediated through NPCE EVs surface proteins in an active manner that involves endocytosis-dependent routes.

The decline of Fraxinus angustifolia Vahl in a Mediterranean salt meadow: Chlorophyll fluorescence measurements in long-term field experiment

The ecological transition zones also known as ecotones are specific ecosystems but how global change will affect their biodiversity and functioning is poorly understood. In the case of the areas at the interface between riparian forest and salt meadows the biodiversity conservation remains a major issue. The aim of this work was to evaluate the effects of a salty environment on Fraxinus angustifolia a non-halophyte species. For this, the photochemistry of PSII was in situ explored during the summer between 2007 and 2016. The predawn maximum quantum efficiency of PSII (Fv/Fm) was always greater than 0.8 which indicates an absence of chronic photoinhibition induced by salinity. During these years, the light kinetic of the actual quantum yield of PSII (ΦPSII) was studied. It appears that this yield is generally higher in salty environments than in the riparian forest. Multivariate analysis, showed that the dissipation regulated light energy faction (ΦNPQ), the photochemical quenching (qP) and the Fd/Fs ratio constitute the main explanatory variables of ΦPSII changes. The subsequent two-way-ANCOVA carried out demonstrated that ΦNPQ was weaker in saline environments, suggesting a better efficiency of photosystem collecting antennas. Meanwhile, the analysis of photosynthetic pigments did not exhibited a salt effect except the significant accumulation of chlorophyll a in 2016. In order to address the question of the salt-related toxicity, sodium ion contents in the leaves were found to be identical in the two sites, suggesting the establishment of an energy-dependent mechanism of salt exclusion/retention in roots. The growth rings of F. angustifolia in saline areas highly follows the annual water deficit changes. Between 2007 and 2016, the biomass in the salt meadow was higher with a peak in 2011 representing 60% of the biomass of control individuals. But, since 2011 a continuous decline of growth was recorded. Water conditions seem to constitute a limiting factor even if the photosynthetic apparatus generates the excess metabolic energy necessary to cope with the saline toxicity. This study thus highlighted the plasticity of the photochemistry of the PSII depending to the environmental conditions of the salt meadows. It also shows the vulnerability of an ecotone through the decline in the productivity of a glycophyte with climate change. This augurs for changes in the composition of plant communities which is in favor of strict halophytes.

Large-scale simulations of antihelium production in cosmic-ray interactions

The possibility of antihelium production in interaction of cosmic rays with the interstellar gas is studied using large-scale Monte Carlo simulations. For this purpose, an energy-dependent coalescence mechanism developed previously is extended to estimate the production of light antinuclei (${}^3\overline{\text{He}}$ and ${}^4\overline{\text{He}}$). The uncertainty in the coalescence parameter and its effect on the expected antiparticle flux is also investigated. The simulated background antihelium fluxes are found to be lower than the fluxes predicted by simplified models using numerical scaling techniques.

Imidazole and Benzimidazole Modified Half-Sandwich IridiumIII N-Heterocyclic Carbene Complexes: Synthesis, Anticancer Application, and Organelle Targeting.

Herein, we report the synthesis, characterization and anticancer activity of a series of half-sandwich iridiumIII imidazole and benzimidazole N-heterocyclic carbene (NHC) anticancer complexes, and the general formula of which can be expressed as [(η5-Cpx)Ir(C∧N)Cl]Cl (Cpx: pentamethylcyclopentadienyl (Cp*) or biphenyl derivatives (Cpxbiph); C∧N: imidazole and benzimidazole NHC chelating ligands). Compared with cis-platin, these complexes showed interesting antitumor activity against A549 cells. Complexes could bind to bovine serum albumin (BSA) by means of static quenching mode, catalyze the oxidation of nicotinamide adenine dinucleotide (NADH) and increase the levels of reactive oxygen species (ROS). Meanwhile, these complexes could arrest the cell cycles of A549 cells and influence the mitochondrial membrane potential significantly. Due to the inherent luminescence property, laser confocal test show that complexes could enter cells followed an energy-dependent mechanism and effectively accumulate in lysosome (the value of Pearson's co-localization coefficient is 0.70 after 1 h), further destroy lysosome integrity and induce apoptosis.

Opportunistic Use by <i>Pseudomonas</i> Aeruginosa of Catecholamine Neurotransmitters as Siderophore to Access Iron

Iron is an essential nutrient for bacteria growth and microorganisms have developped several strategies to access iron, the most common being the production of siderophores, iron-chelating molecules secreted into the bacterial environment. Here, we demonstrate that catecholamine neurotransmitters (dopamine, L-DOPA, epinephrine, and norepinephrine) are able to chelate iron and bring iron into Pseudomonas aeruginosa cells via an energy-dependent transport mechanism that relies on TonB-dependent transporters (TBDTs). Using bacterial growth assay under strong iron-restricted conditions, we show that the TBDTs involved are PiuA and PirA. PiuA exhibited more pronounced specificity for dopamine uptake, whereas PirA specificity appeared to be higher for L-DOPA and norepinephrine. Proteomic and qRT-PCR approaches showed pirA transcription and expression to be induced in the presence of all four catecholamines. Finally, the oxidative properties of catecholamines enable them to reduce iron, and we observed ferrous iron uptake via the FeoABC system in the presence of L-DOPA.

Fluorescent iridium(iii) coumarin-salicylaldehyde Schiff base compounds as lysosome-targeted antitumor agents.

Six fluorescent half-sandwich iridium(iii) coumarin-salicylaldehyde Schiff base (O^N) compounds ([(η5-Cp*)Ir(O^N)Cl]) were prepared and characterized. The introduction of a coumarin unit increased the antitumor activity (IC50: 9.9 ± 0.1 μM-40.7 ± 12.9 μM) of these compounds, the best of which was nearly two times that of clinical cisplatin. The results of laser confocal microscopy demonstrated that these compounds possessed an energy-dependent cellular uptake mechanism, accumulated in the lysosomes (Pearson co-localization coefficient: ∼0.7), damaged the integrity of the lysosomes, and induced apoptosis. The compounds could also decrease the mitochondrial membrane potential, catalyze the oxidation of the coenzyme (nicotinamide-adenine dinucleotide) and improve the levels of the intracellular reactive oxygen species, following an antitumor mechanism of oxidation. Additionally, these compounds could block the metastasis of tumor cells. Above all, these iridium(iii) compounds show potential as antitumor agents with dual functions: lysosomal damage and anti-metastasis.

Synthesis of terpolymer-lipid encapsulated diruthenium(II,III)-anti-inflammatory metallodrug nanoparticles to enhance activity against glioblastoma cancer cells

Novel formulations of diruthenium(II,III)-NSAID (NSAID, non-steroidal anti-inflammatory drug) metallodrugs encapsulated into biocompatible terpolymer-lipid nanoparticles (TPLNs) to target glioblastoma cancer were developed. The nanoformulations of Ibuprofenate (RuIbp) and Naproxenate (RuNpx) metallodrugs were synthesized and characterized. The procedure rationally designed to avoid structural changes on the coordination sphere of the [Ru2(NSAID)4]+ paddlewheel unit succeeded in giving colloidally stable and nearly spherical shaped loaded [Ru2(NSAID)4]-TPLNs with appropriate parameters (~90% loading efficiency; drug loading around 10%; particle size ~130 nm; zeta potential around - 40 mV). The maintenance of the [Ru2(NSAID)4]+ framework was confirmed by spectroscopy and mass spectrometry. The encapsulation enhanced antiproliferative effects in U87MG cells for both metallodrugs. The RuIbp-TPLNs showed efficacy also against the cisplatin chemoresistant T98G cancer cells. Lack of significant effects for the loaded-Ibuprofen-TPLNs (HIbp-TPLNs) on both types of cells supports the key role of the dimetal core in the anticancer activity of the [Ru2(NSAID)4]+ metallodrugs. The high cell viability (>70%) found for both types of cells suggests activity associated mainly to antiproliferative effects. The blank-TPLNs internalized into U87MG cell cytoplasm mostly at the first 6 h, by energy-dependent mechanism. The cell uptake of the RuIbp-TPLNs occurred during the first 24 h and it was enhanced in relation to the non-encapsulated metallodrug. The development of these novel metallodrug-loaded TPLN nanoformulations, which exhibit colloidal stability suitable for intravenous injection and enhanced drug cellular uptake, expands the perspective for diruthenium(II,III)-NSAID metallodrugs targeting brain glioblastoma cancer.