Lemon essential oil nanoemulsions: Potential natural inhibitors against Escherichia coli.

Lemon essential oil (LEO) is a common natural antibacterial substance, and encapsulating LEO into nanoemulsions (NEs) can improve their stability and broaden its application. Our study aimed to investigate the bacterial inhibitory effect of LEO-NEs against Escherichia coli (E. coli). Results showed that the minimum inhibitory concentration (MIC) of LEO-NEs was 6.25 mg/mL, and the time-kill curve showed that E. coli were significantly killed by LEO-NEs after 5 h of treatment at 1MIC. Flow-cytometry analysis showed that LEO-NEs adversely affected the cell-membrane depolarisation, cell-membrane integrity, and efflux pump function of E. coli. Confocal laser scanning microscopy demonstrated that 8MIC of LEO-NEs induced changes in the cell-membrane permeability and cell-wall integrity of E. coli. Proteomic results suggested that the mode of action LEO-NEs against E. coli was to enhance bacterial chemotaxis and significantly inhibit ribosomal assembly. They may also affect butyric acid, ascorbic acid and aldehyde metabolism, and sulphur-relay system pathways. In conclusion, LEO-NEs had potential application as a natural antibacterial agent for the control of E. coli in the food industry.

Linalool controls the viability of Escherichia coli by regulating the synthesis and modification of lipopolysaccharide, the assembly of ribosome, and the expression of substrate transporting proteins.

Escherichia coli (E. coli) is a Gram-negative bacterium and some pathogenic types may cause serious diseases, foods or food environments were the primary routes for its infection. Citrus aurantium L. var. amara Engl., a variety of sour orange, were used as a kind of non-conventional edible plant in China, but its antimicrobial activity and mechanisms were not well studied. Thus, in this study, EO from the flower of Citrus aurantium L. var. amara Engl. (CAEO) were studied as a kind of natural antimicrobial agent to control E. coli, our results showed that both of CAEO and its main component (linalool) exhibited strong antibacterial efficacy. Further, transcriptomic and proteomic analysis were carried out to explore cell response under linalool treatment and the main results included: (1) The synthesis and modification of lipopolysaccharide (LPS) was significantly influenced. (2) Ribosomal assembly and protein synthesis were significantly inhibited. (3) The expression of proteins related to the uptake of several essential substances was significantly changed. In all, our results would supply a theoretical basis for the proper use of CAEO and linalool as a promising antimicrobial agent to prevent and control E. coli infection in the future.

Essential Oils From Citrus unshiu Marc. Effectively Kill Aeromonas hydrophila by Destroying Cell Membrane Integrity, Influencing Cell Potential, and Leaking Intracellular Substances.

Aeromonas hydrophila is one of the important pathogenic bacteria in aquaculture causing serious losses every year. Essential oils are usually used as natural antimicrobial agents to reduce or replace the use of antibiotics. The aim of this study was to evaluate the antibacterial activity and explore the mechanisms of essential oil from satsuma mandarin (Citrus unshiu Marc.) (SMEO) against A. hydrophila. The results of the gas chromatography-mass spectrometer demonstrated that SMEO contains 79 chemical components with the highest proportion of limonene (70.22%). SMEO exhibited strong antibacterial activity against A. hydrophila in vitro, the diameter of the inhibition zone was 31.22 ± 0.46 mm, and the MIC and MBC values were all 1% (v/v). Intracellular material release, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and flow cytometry analysis revealed the dynamic antibacterial process of SMEO, the morphological changes of bacterial cells, and the leakage process of intracellular components. These results demonstrated that SMEO disrupted the extracellular membrane permeability. Our study demonstrated that SEMO has the potential to be used to control and prevent A. hydrophila infections in aquaculture.

Reassessment of long circulation via monitoring of integral polymeric nanoparticles justifies a more accurate understanding.

Monitoring of payloads results in a biased perception of long circulation of nanoparticles. Instead, herein, the long-circulation effect was re-confirmed by monitoring integral nanoparticles, but circulation was not found to be as long as generally perceived. In contrast, disparate pharmacokinetics were obtained by monitoring a model drug, paclitaxel, highlighting the bias of the conventional protocol.

Correction: Reassessment of long circulation via monitoring of integral polymeric nanoparticles justifies a more accurate understanding.

Correction for 'Reassessment of long circulation via monitoring of integral polymeric nanoparticles justifies a more accurate understanding' by Haisheng He et al., Nanoscale Horiz., 2018, DOI: .

Influence of Particle Geometry on Gastrointestinal Transit and Absorption following Oral Administration.

Geometry has been considered as one of the important parameters in nanoparticle design because it affects cellular uptake, transport across the physiological barriers, and in vivo distribution. However, only a few studies have been conducted to elucidate the influence of nanoparticle geometry in their in vivo fate after oral administration. This article discloses the effect of nanoparticle shape on transport and absorption in gastrointestinal (GI) tract. Nanorods and nanospheres were prepared and labeled using fluorescence resonance energy transfer molecules to track the in vivo fate of intact nanoparticles accurately. Results demonstrated that nanorods had significantly longer retention time in GI tract compared with nanospheres. Furthermore, nanorods exhibited stronger ability of penetration into space of villi than nanospheres, which is the main reason of longer retention time. In addition, mesenteric lymph transported 1.75% nanorods within 10 h, which was more than that with nanospheres (0.98%). Fluorescent signals arising from nanoparticles were found in the kidney but not in the liver, lung, spleen, or blood, which could be ascribed to low absorption of intact nanoparticles. In conclusion, nanoparticle geometry influences in vivo fate after oral delivery and nanorods should be further investigated for designing oral delivery systems for therapeutic drugs, vaccines, or diagnostic materials.

Size-Dependent Translocation of Nanoemulsions via Oral Delivery.

The in vivo translocation of nanoemulsions (NEs) was tracked by imaging tools with an emphasis on the size effect. To guarantee the accurate identification of NEs in vivo, water-quenching environment-responsive near-infrared fluorescent probes were used to label NEs. Imaging evidence confirmed prominent digestion in the gastrointestinal tract and oral absorption of integral NEs that survive digestion by enteric epithelia in a size-dependent way. In general, reducing particle size leads to slowed in vitro lipolysis and in vivo digestion, a prolonged lifetime in the small intestine, increased enteric epithelial uptake, and enhanced transportation to various organs. Histological examination revealed a pervasive distribution of smaller NEs (80 nm) into enterocytes and basolateral tissues, whereas bigger ones (550, 1000 nm) primarily adhered to villi surfaces. Following epithelial uptake, NEs are transported through the lymphatics with a fraction of approximately 3-6%, suggesting a considerable contribution of the lymphatic pathway to overall absorption. The majority of absorbed NEs were found 1 h post administration in the livers and lungs. A similar size dependency of cellular uptake and transmonolayer transport was confirmed in Caco-2 cell lines as well. In conclusion, the size-dependent translocation of integral NEs was confirmed with an absolute bioavailability of at least 6%, envisioning potential applications in oral delivery of labile entities.

Transmembrane delivery of anticancer drugs through self-assembly of cyclic peptide nanotubes.

Breaking the natural barriers of cell membranes achieves fast entry of therapeutics, which leads to enhanced efficacy and helps overcome multiple drug resistance. Herein, transmembrane delivery of a series of small molecule anticancer drugs was achieved by the construction of artificial transmembrane nanochannels formed by self-assembly of cyclic peptide (cyclo[Gln-(d-Leu-Trp)4-d-Leu], CP) nanotubes (CPNTs) in the lipid bilayers. Our in vitro study in liposomes indicated that the transport of molecules with sizes smaller than 1.0 nm, which is the internal diameter of the CPNTs, could be significantly enhanced by CPNTs in a size-selective and dose-dependent manner. Facilitated uptake of 5-fluorouracil (5-FU) was also confirmed in the BEL7402 cell line. On the contrary, CPs could facilitate neither the transport across liposomal membranes nor the uptake by cell lines of cytarabine, a counterevidence drug with a size of 1.1 nm. CPs had a very weak anticancer efficacy, but could significantly reduce the IC50 of 5-FU in BEL7402, HeLa and S180 cell lines. Analysis by a q test revealed that a combination of 5-FU and CP had a synergistic effect in BEL7402 at all CP levels, in S180 at CP levels higher than 64 µg mL(-1), but not in HeLa, where an additive effect was observed. Temporarily, intratumoral injection is believed to be the best way for CP administration. In vivo imaging using (125)I radio-labelled CP confirmed that CPNPTs were completely localized in the tumor tissues, and translocation to other tissues was negligible. In vivo anticancer efficacy was studied in the grafted S180 solid tumor model in mice, and the results indicated that tumor growth was greatly inhibited by the combinatory use of 5-FU and CP, and a synergistic effect was observed at CP doses of 0.25 mg per kg bw. It is concluded that facilitated transmembrane delivery of anticancer drugs with sizes smaller than 1.0 nm was achieved, and the synergistic anticancer effect was confirmed both in cell lines and in vivo through the combinatory use of 5-FU and CP.

Glucan microparticles thickened with thermosensitive gels as potential carriers for oral delivery of insulin.

Although glucan microparticles (GMs) can be efficiently taken up and transported by M cells, their subsequent accumulation in lymphatic tissues of sub-follicle-associated epithelia (FAE) in Peyer's patches might present a barrier to the oral delivery of insulin by GMs into the systemic circulation. The goal of this study is to weigh the potential of GMs as carriers for oral delivery of systemic therapeutics using insulin (INS) as a model drug. INS is encapsulated into the inner cavities of GMs by repeated soaking in INS solution at acidic pH values and switching to an isoelectric pH of 5.6 to precipitate INS. To immobilize INS, a thermosensitive poloxamer 407 (P407) gel is introduced into the interior of GMs. Interiorly thickened GMs show significantly decreased in vitro release and well protected INS stability against enzyme-enriched media, highlighting the importance of thickening with P407 gels. A mild and prolonged hypoglycaemic effect is achieved in both normal and diabetic rats for a duration of at least 20 h with pharmacological bioavailability as high as about 9-10%. Lymphatic transportation of GMs is investigated by labelling with a near-infrared water-quenching fluorescent probe in a conscious mesentery lymphatic duct cannulation rat model following oral administration. GMs appear in lymph within the first 2 h, peak at around 6 h and slow down after 10 h with a cumulative amount of over 8% in 24 h. The high correlation between lymphatic transportation and pharmacological bioavailability implies that GMs are principally absorbed via the lymphatic route. An in vitro study on phagocytosis by macrophages confirms the easy and fast uptake of GMs by J774A.1 cell lines with as many as over 10 particles within the cytoplasm of a single cell. Intracellular pharmacokinetics indicates robustness and persistent residence of GMs within the cells. Little effect on cell viability and tight junctions was observed in Caco-2 cell models. It is concluded that GMs are mainly absorbed via the lymphatic route and show potential as carriers for oral delivery of labile therapeutics, though with limited bioavailability due to the sub-FAE residence barriers.