Chemo-optogenetic Protein Translocation System Using a Photoactivatable Self-Localizing Ligand.

Manipulating subcellular protein localization using light is a powerful approach for controlling signaling processes with high spatiotemporal precision. The most widely used strategy for this is based on light-induced protein heterodimerization. The use of small synthetic molecules that can control the localization of target proteins in response to light without the need for a second protein has several advantages. However, such methods have not been well established. Herein, we present a chemo-optogenetic approach for controlling protein localization using a photoactivatable self-localizing ligand (paSL). We developed a paSL that can recruit tag-fused proteins of interest from the cytoplasm to the plasma membrane within seconds upon light illumination. This paSL-induced protein translocation (paSLIPT) is reversible and enables the spatiotemporal control of signaling processes in living cells, even in a local region. paSLIPT can also be used to implement simultaneous optical stimulation and multiplexed imaging of molecular processes in a single cell, offering an attractive and novel chemo-optogenetic platform for interrogating and engineering dynamic cellular functions.

Photodegradation of Ciprofloxacin, Clarithromycin and Trimethoprim: Influence of pH and Humic Acids.

In view of the rising relevance of emerging pollutants in the environment, this work studies the photodegradation of three antibiotics, evaluating the effects of the pH of the medium and the concentration of dissolved organic matter. Simulated light (with a spectrum similar to that of natural sunlight) was applied to the antibiotics Ciprofloxacin (Cip), Clarithromycin (Cla) and Trimethoprim (Tri), at three different pH, and in the presence of different concentrations of humic acids. The sensitivity to light followed the sequence: Cip > Cla > Tri, which was inverse for the half-life (Tri > Cla > Cip). As the pH increased, the half-life generally decreased, except for Cla. Regarding the kinetic constant k, in the case of Cip and Tri it increased with the rise of pH, while decreased for Cla. The results corresponding to total organic carbon (TOC) indicate that the complete mineralization of the antibiotics was not achieved. The effect of humic acids was not marked, slightly increasing the degradation of Cip, and slightly decreasing it for Tri, while no effect was detected for Cla. These results may be relevant in terms of understanding the evolution of these antibiotics, especially when they reach different environmental compartments and receive sunlight radiation.

Influence of ionizing radiation on the antimicrobial activity of the antibiotics sulfamethoxazole and trimethoprim.

The response of the antimicrobial compounds sulfamethoxazole (SMX) and trimethoprim (TMP) - individually and in mixtures - to ionizing radiation was investigated using laboratory prepared mixtures and a commercial pharmaceutical formulation. The residual antibacterial activity of the solutions was monitored using Staphylococcus aureus and Escherichia coli test strains. Based on antibacterial activity, SMX was more susceptible to ionizing radiation as compared to TMP. The antibacterial activity of SMX and TMP was completely eliminated at 0.2 kGy and 0.8 kGy, respectively. However, when SMX and TMP were in a mixture, the dose required to eliminate the antibacterial activity was 10 kGy, implying a synergistic antibacterial activity when these are present in mixtures. Only when the antibiotic concentration was below the Minimum Inhibitory Concentration of TMP (i.e., 2 µmol dm-3) did the antibacterial activity of the SMX and TMP mixture disappear. These results imply that the synergistic antimicrobial activity of antimicrobial compounds in pharmaceutical waste streams is a strong possibility. Therefore, antimicrobial activity assays should be included when evaluating the use of ionizing radiation technology for the remediation of pharmaceutical or municipal waste streams.

Decomposition of sulfamethoxazole and trimethoprim by continuous UVA/LED/TiO2 photocatalysis: Decomposition pathways, residual antibacterial activity and toxicity.

In this study, continuous LED/UVA/TiO2 photocatalytic decomposition of sulfamethoxazole (SMX) and trimethoprim (TMP) was investigated. More than 90% of SMX and TMP were removed within 20min by the continuous photoreactor (with the initial concentration of 400ppb for each). The removal rates of SMX and TMP decreased with higher initial antibiotics loadings. SMX was much easier decomposed in acidic condition, while pH affected little on TMP's decomposition. 0.003% was found to be the optimum H2O2 dosage to enhance SMX photocatalytic decomposition. Decomposition pathways of SMX and TMP were proposed based on the intermediates identified by using LC-MS-MS and GC-MS. Aniline was identified as a new intermediate generated during SMX photocatalytic decomposition. Antibacterial activity study with a reference Escherichia coli strain was also conducted during the photocatalytic process. Results indicated that with every portion of TMP removed, the residual antibacterial activity decreased by one portion. However, the synergistic effect between SMX and TMP tended to slow down the antibacterial activity removal of SMX and TMP mixture. Chronic toxicity studies conducted with Vibrio fischeri exhibited 13-20% bioluminescence inhibition during the decomposition of 1ppm SMX and 1ppm TMP, no acute toxicity to V. fischeri was observed during the photocatalytic process.

Removal of trimethoprim, sulfamethoxazole, and triclosan by the green alga Nannochloris sp.

Trimethoprim (TMP), sulfamethoxazole (SMX), and triclosan (TCS) are widely used and continuously released into aquatic environments. Freshwater algae can be responsible for the uptake and transfer of the contaminants because they are a major food source for most aquatic organisms. This research applied incubation studies to evaluate the removal efficiency of TMP, SMX, and TCS by the green alga Nannochloris sp. The results showed that the hydrophilic antibiotics TMP and SMX remained in the algal culture at 100% and 68%, respectively, after 14days of incubation, and therefore were not significantly removed from the medium. However, the lipophilic antimicrobial TCS was significantly removed from the medium. Immediately after incubation began, 74% of TCS dissipated and 100% of TCS was removed after 7days of incubation. Additionally, over 42% of TCS was found associated with the algal cells throughout the incubation. The results demonstrate that the presence of Nannochloris sp. eliminated TCS in the aquatic system, but could not significantly remove the antibiotics TMP and SMX. The removal mechanisms of SMX and TCS were found to be different in the algal culture. Algae-promoted photolysis was the primary process for removing SMX and algae-mediated uptake played a major role in removing TCS.

Photolytic degradation of sulfamethoxazole and trimethoprim using UV-A, UV-C and vacuum-UV (VUV).

The photolytic degradation of the non-degradable pharmaceuticals sulfamethoxazole (SMX) and trimethoprim (TMP) in an aqueous solution was investigated using three kinds of low-pressure mercury lamp UV-A (352 nm), UV-C (254 nm), and vacuum-UV (VUV, 185 nm and 254 nm). The degradation rates were highly dependent on the target compounds as well as the UV sources. No degradation of the target compounds was observed using UV-A treatment, because there was no overlap between the UV-A emission spectrum and absorption spectrum of the target compounds. On the other hand, UVC and VUV revealed higher reactivity. The results also indicated that SMX had a greater potential to react photochemically than TMP. Among the UV sources, VUV was the most effective process for the degradation of target compounds. Furthermore, the addition of oxidants such as hydrogen peroxide (H2O2) and sodium persulfate (Na2S2O8) to the reaction system improved the overall degradation rate significantly.The experimental results for the VUV-irradiated samples with the addition of methanol as a hydroxyl radical scavenger revealed that hydroxyl radicals contribute significantly to the elimination of the target compound. Overall, the degradation rate of the target compounds was in the order: VUV = UV-C > UV-A for sulfamethoxazole and VUV/H2O2 > VUV/ Na2S2O8 > VUV >UV-C >UV-A for trimethoprim.

Superiority of solar Fenton oxidation over TiO2 photocatalysis for the degradation of trimethoprim in secondary treated effluents.

The overall aim of this work was to examine the degradation of trimethoprim (TMP), which is an antibacterial agent, during the application of two advanced oxidation process (AOP) systems in secondary treated domestic effluents. The homogeneous solar Fenton process (hv/Fe(2+)/H2O2) and heterogeneous photocatalysis with titanium dioxide (TiO2) suspensions were tested. It was found that the degradation of TMP depends on several parameters such as the amount of iron salt and H2O2, concentration of TiO2, pH of solution, solar irradiation, temperature and initial substrate concentration. The optimum dosages of Fe(2+) and H2O2 for homogeneous ([Fe(2+)] = 5 mg L(-1), [H2O2] = 3.062 mmol L(-1)) and TiO2 ([TiO2] = 3 g L(-1)) for heterogeneous photocatalysis were established. The study indicated that the degradation of TMP during the solar Fenton process is described by a pseudo-first-order reaction and the substrate degradation during the heterogeneous photocatalysis by the Langmuir-Hinshelwood kinetics. The toxicity of the treated samples was evaluated using a Daphnia magna bioassay and was finally decreased by both processes. The results indicated that solar Fenton is more effective than the solar TiO2 process, yielding complete degradation of the examined substrate within 30 min of illumination and dissolved organic carbon (DOC) reduction of about 44% whereas the respective values for the TiO2 process were ∼70% degradation of TMP within 120 min of treatment and 13% DOC removal.

Effect of water-matrix composition on Trimethoprim solar photodegradation kinetics and pathways.

Direct photolysis and solar TiO(2) photocatalysis of Trimethoprim (TMP) in different water matrices (demineralised and simulated seawater) have been studied. Direct photolysis yielded a similar, slow TMP degradation rate in both water matrices, and the formation of very stable photo-transformation products. Dissolved organic carbon decreased slightly after prolonged irradiation. The main intermediate identified was a ketone derivative (trimethoxybenzoylpyrimidine), which was proved to be a photosensitizer of TMP degradation. During TiO(2) photocatalysis, TMP was completely eliminated in both water matrices at a similar rate, however, the mineralization rate was appreciably reduced in seawater, which can be explained by the presence of inorganic species acting as hydroxyl radical scavengers, and directly affecting photocatalytic efficiency. Identification of intermediates showed differences between the two processes but hydroxylation, demethylation and cleavage of the original drug molecule were observed in both.