Bacteriomimetic Liposomes Improve Antibiotic Activity of a Novel Energy-Coupling Factor Transporter Inhibitor

Liposomes have been studied for decades as nanoparticulate drug delivery systems for cytostatics, and more recently, for antibiotics. Such nanoantibiotics show improved antibacterial efficacy compared to the free drug and can be effective despite bacterial recalcitrance. In this work, we present a loading method of bacteriomimetic liposomes for a novel, hydrophobic compound (HIPS5031) inhibiting energy-coupling factor transporters (ECF transporters), an underexplored antimicrobial target. The liposomes were composed of DOPG (18:1 (Δ9-cis) phosphatidylglycerol) and CL (cardiolipin), resembling the cell membrane of Gram-positive Staphylococcus aureus and Streptococcus pneumoniae, and enriched with cholesterol (Chol). The size and polydispersity of the DOPG/CL/± Chol liposomes remained stable over 8 weeks when stored at 4 °C. Loading of the ECF transporter inhibitor was achieved by thin film hydration and led to a high encapsulation efficiency of 33.19% ± 9.5% into the DOPG/CL/Chol liposomes compared to the phosphatidylcholine liposomes (DMPC/DPPC). Bacterial growth inhibition assays on the model organism Bacillus subtilis revealed liposomal HIPS5031 as superior to the free drug, showing a 3.5-fold reduction in CFU/mL at a concentration of 9.64 µM. Liposomal HIPS5031 was also shown to reduce B. subtilis biofilm. Our findings present an explorative basis for bacteriomimetic liposomes as a strategy against drug-resistant pathogens by surpassing the drug-formulation barriers of innovative, yet unfavorably hydrophobic, antibiotics.

INTERACTION OF 3-HYDROXY PYRIDINE AND SURFACTANT MICELLES: A FLUORESCENCE STUDIES

Objective: Micellar solubilization is a powerful alternative for dissolving hydrophobic compound in aqueous environment. 3-hydroxy pyridine (3- HP) derivatives are the potential endogenous photosensitizers. 3-HP derivatives show protective effect in clinical extreme condition such as hypoxia, hyperthermia, hypokinesia. Micellization of 3-HP followed by solubilization would catalyze its pharmaceutical activities which may serve better results in medicinal and analytical fields. Methods: Fluorescence and absorption spectroscopy techniques are used to monitor the micellar solubilization studies of 3-HP. Solubilization studies of 3-HP with various anionic, cationic and nonionic surfactants have been performed in aqueous medium around 23–25°C temperature. The solubilization action of the surfactant has also been determined by theoretical calculated spectral parameters such as empirical fluorescence coefficient, quantum yield, stokes, shift and molar absorption coefficient. Results: 3-HP shows fluorescence excitation peak at 315 nm and emission peak at 390 nm respectively while the absorbance of 3-HP has been found to be maximum at 305 nm. The fluorescence as well as the theoretically calculated spectral data has been used to characterize the hetero environment of the micelles in terms of their polarity, probe solubilization site and critical micelle concentration. Conclusion: This article briefly discusses the importance of surfactants in biological system model as well as the use of micelles in pharmacy as an important tool that finds numerous applications.

Anaerobic Co-Digestion Effluent as Substrate for Chlorella vulgaris and Scenedesmus obliquus Cultivation

Anaerobic digestate supernatant can be used as a nutrient source for microalgae cultivation, thus integrating phytoremediation processes with high value products storage in microalgae biomass. Microalgae are able to use nitrogen and phosphorous from digestate, but high nutrient concentration can cause growth inhibition. In this study, two microalgae strains (C. vulgaris and S. obliquus) were cultivated on the anaerobic co-digestion supernatant (obtained from the organic fraction of municipal solid waste (OFMSW) and waste activated sludge (WAS)) in a preliminary Petri plate screening at different dilutions (1:10 and 1:5) using a synthetic medium (ISO) and tap water (TW). Direct Nile red screening was applied on colonies to preliminarily identify hydrophobic compound storage and then a batch test was performed (without air insufflation). Results show that C. vulgaris was able to grow on digestate supernatant 1:5 diluted, while Nile red screening allowed the preliminary detection of hydrophobic compound storage in colonies. The analysis carried out at the end of the test on ammonia, phosphate, nitrate and sulphate showed a removal percentage of 47.5 ± 0.8%, 65.0 ± 6.0%, 95.0 ± 3.0% and 99.5 ± 0.1%, respectively.

Lipophilicity of Bacteriochlorin-Based Photosensitizers as a Determinant for PDT Optimization through the Modulation of the Inflammatory Mediators

Photodynamic therapy (PDT) augments the host antitumor immune response, but the role of the PDT effect on the tumor microenvironment in dependence on the type of photosensitizer and/or therapeutic protocols has not been clearly elucidated. We employed three bacteriochlorins (F2BOH, F2BMet and Cl2BHep) of different polarity that absorb near-infrared light (NIR) and generated a large amount of reactive oxygen species (ROS) to compare the PDT efficacy after various drug-to-light intervals: 15 min. (V-PDT), 3h (E-PDT) and 72h (C-PDT). We also performed the analysis of the molecular mechanisms of PDT crucial for the generation of the long-lasting antitumor immune response. PDT-induced damage affected the integrity of the host tissue and developed acute (protocol-dependent) local inflammation, which in turn led to the infiltration of neutrophils and macrophages. In order to further confirm this hypothesis, a number of proteins in the plasma of PDT-treated mice were identified. Among a wide range of cytokines (IL-6, IL-10, IL-13, IL-15, TNF-α, GM-CSF), chemokines (KC, MCP-1, MIP1α, MIP1β, MIP2) and growth factors (VEGF) released after PDT, an important role was assigned to IL-6. PDT protocols optimized for studied bacteriochlorins led to a significant increase in the survival rate of BALB/c mice bearing CT26 tumors, but each photosensitizer (PS) was more or less potent, depending on the applied DLI (15 min, 3 h or 72 h). Hydrophilic (F2BOH) and amphiphilic (F2BMet) PSs were equally effective in V-PDT (>80 cure rate). F2BMet was the most efficient in E-PDT (DLI = 3h), leading to a cure of 65 % of the animals. Finally, the most powerful PS in the C-PDT (DLI = 72 h) regimen turned out to be the most hydrophobic compound (Cl2BHep), allowing 100 % of treated animals to be cured at a light dose of only 45 J/cm2.

Storing and releasing rhodamine as a model hydrophobic compound in polydimethylsiloxane microfluidic devices

Polydimethylsiloxane (PDMS) is known to absorb small hydrophobic molecules. We propose to leverage this material to store and release small hydrophobic molecules into and from the PDMS matrix. This method could be used to deliver small hydrophobic molecules to microfluidic channels from the walls of a microfluidic device.