Macromolecular antimicrobial agents such as cationic polymers and peptides have recently been under an increased level of scrutiny because they can combat multi-drug-resistant microbes. Most of these polymers are non-biodegradable and are designed to mimic the facially amphiphilic structure of peptides so that they may form a secondary structure on interaction with negatively charged microbial membranes. The resulting secondary structure can insert into and disintegrate the cell membrane after recruiting additional polymer molecules. Here, we report the first biodegradable and in vivo applicable antimicrobial polymer nanoparticles synthesized by metal-free organocatalytic ring-opening polymerization of functional cyclic carbonate. We demonstrate that the nanoparticles disrupt microbial walls/membranes selectively and efficiently, thus inhibiting the growth of Gram-positive bacteria, methicillin-resistant Staphylococcus aureus (MRSA) and fungi, without inducing significant haemolysis over a wide range of concentrations. These biodegradable nanoparticles, which can be synthesized in large quantities and at low cost, are promising as antimicrobial drugs, and can be used to treat various infectious diseases such as MRSA-associated infections, which are often linked with high mortality.

Non-viral gene-delivery systems are safer to use and easier to produce than viral vectors, but their comparatively low transfection efficiency has limited their applications. Co-delivery of drugs and DNA has been proposed to enhance gene expression or to achieve the synergistic/combined effect of drug and gene therapies. Attempts have been made to deliver drugs and DNA simultaneously using liposomes. Here we report cationic core-shell nanoparticles that were self-assembled from a biodegradable amphiphilic copolymer. These nanoparticles offer advantages over liposomes, as they are easier to fabricate, and are more readily subject to modulation of their size and degree of positive charge. More importantly, they achieve high gene-transfection efficiency and the possibility of co-delivering drugs and genes to the same cells. Enhanced gene transfection with the co-delivery of paclitaxel has been demonstrated by in vitro and in vivo studies. In particular, the co-delivery of paclitaxel with an interleukin-12-encoded plasmid using these nanoparticles suppressed cancer growth more efficiently than the delivery of either paclitaxel or the plasmid in a 4T1 mouse breast cancer model. Moreover, the co-delivery of paclitaxel with Bcl-2-targeted small interfering RNA (siRNA) increased cytotoxicity in MDA-MB-231 human breast cancer cells.

Quantum dots (QDs) are of great interest to biological applications such as fluorescent biosensors and biolabels. Our study describes a synthesis of glutathione-capped CdTe QDs in aqueous solution that is cost-efficient and convenient compared to the conventional organometallic approaches. The fluorescence of the as-prepared glutathione-capped CdTe QDs was tunable from 500 to 650 nm. Without any postpreparation treatment, the glutathione-capped CdTe QDs achieved quantum yields (QYs) as high as 45 %, comparable to, or even better than, QDs derived from organometallic routes. With an overall size as small as 4 nm, they could gain access to cellular targets and stain fine features such as the cell nucleolus. These QDs were successfully conjugated with biotin for immunostaining and with F3 peptide for delivery to live cells, demonstrating their potentially broad application as biolabels.Fluorescent semiconductor nanoparticles or QDs have been extensively investigated in the past decade, and have been widely used as biolabels in imaging and biodetection.[1] Fluorescence imaging can greatly benefit from the use of QDs, which show brighter fluorescence, less photobleaching, and multiple colors with a single excitation. For biolabeling applications, QDs are commonly capped with trioctylphosphine oxide (TOPO) through organometallic synthesis, followed by phospholipid, [2] silica, [3] or polymer [4] coating to impart watersolubility and biocompatibility. With the multilayer coating, the final size of the QD biolabels would typically be 12-20 nm, which might be too bulky to gain access to cells for in vitro and in vivo imaging. In addition, the large dimensions would dramatically lower the labeling efficiency on specific target sites within the cells. Compared to conventionally used organic dyes, the size of QDs presented a major drawback as it limited the range of applications in biolabeling. To overcome this obstacle, water-soluble QDs with only one layer of capping ligand on the surface have been developed by ligand exchange with thiols [5] or phosphine [6] on TOPO-capped QDs.Both the ligand-addition and ligand-exchange methods were complicated, and even the high-quality QDs derived from organometallic methods have exhibited reductions in QY from 65-85 % in the organic phase to 35-50 % in aqueous solution.[7]Alternatively, thiol-capped QDs could be prepared directly in aqueous solution with thiols as stabilizers, but low QYs of 1-10 % were typically obtained.[8] Although their QYs could be significantly improved by a variety of after-treatments, such as photochemical etching, [8] size-selective precipitation, [9] and long-term illumination, [10] these QDs have a tendency to agglomerate during such treatments. Glutathione is a thiol-containing oligopeptide found in most organisms, and it plays an important role in the detoxification of heavy metals in plant cells. The physiological mechanism of detoxification involves the binding of heavy-metal nanoclusters by glutathione and the formation of a phytochelatin shell, cat...

Polymyxins remain the last line treatment for multidrug-resistant (MDR) infections. As polymyxins resistance emerges, there is an urgent need to develop effective antimicrobial agents capable of mitigating MDR. Here, we report biodegradable guanidinium-functionalized polycarbonates with a distinctive mechanism that does not induce drug resistance. Unlike conventional antibiotics, repeated use of the polymers does not lead to drug resistance. Transcriptomic analysis of bacteria further supports development of resistance to antibiotics but not to the macromolecules after 30 treatments. Importantly, high in vivo treatment efficacy of the macromolecules is achieved in MDR A. baumannii-, E. coli-, K. pneumoniae-, methicillin-resistant S. aureus-, cecal ligation and puncture-induced polymicrobial peritonitis, and P. aeruginosa lung infection mouse models while remaining non-toxic (e.g., therapeutic index—ED50/LD50: 1473 for A. baumannii infection). These biodegradable synthetic macromolecules have been demonstrated to have broad spectrum in vivo antimicrobial activity, and have excellent potential as systemic antimicrobials against MDR infections.

Nanobiointerfaces were prepared based on an electrically conductive polyethylenedioxythiophene (PEDOT). Thin (<100 nm), ultrasmooth (roughness ( R(rms)) < 5 nm), and functionalized PEDOT films have been successfully electropolymerized using aqueous microemulsion. The microemulsion polymerization is found to be catalyzed in the presence of a low concentration of acid and allows for film formation from various functionalized ethylenedioxythiophenes (EDOTs) (e.g., EDOT-OH, C(2)-EDOT-COOH, C(4)-EDOT-COOH, C(2)-EDOT-NHS, EDOT-N(3)) and their mixtures. The nanobiointerfaces are compositionally tunable and controlled to deposit on selected electrode surfaces. They prefer orthogonal growth on patterned surfaces and are synthesized within seconds. These thin PEDOT films exhibit very low intrinsic cytotoxicity and display no inflammatory response upon implantation, making them ideal for biosensing and bioengineering applications.

Nanoparticle- and quantum-dot (QD)-based bioprobes are emerging as alternatives to small-molecule probes for in vitro and in vivo applications. However, their cellular interaction and cell uptake mechanism are significantly different from those of small-molecule probes and are extremely sensitive to surface ligands. These present a barrier in the development of nanoparticles and QDs as cellular probes. This work focused on the synthesis of various functionalized QDs with tunable surface charge, hydrophobicity, and functionalization with poly(ethylene glycol) (PEGylation) and their cellular interaction. We found that the surface functional groups of nanometer-sized probes significantly dictated their cellular interaction, subcellular localization, and cytotoxicity. A dose-dependent interaction was observed for all types of QDs, but the cationic surface charge or hydrophobicity would increase the cellular interaction as compared to the anionic surface charge. Cationic QDs rapidly entered cells and induced cytotoxicity, but hydrophobic QDs were stuck to the cell membrane and did not enter the cells. PEGylation of cationic QDs reduced their nonspecific binding and cytotoxicity, and a higher concentration of QDs was required for cellular entry. On the basis of these results, we were able to design different functionalized QD nanoprobes with balanced hydrophobicity and surface charge for cell membrane labeling and subcellular targeting. Mechanistic studies indicated a clathrin-mediated interaction and uptake for all types of QDs. The cellular interaction and uptake of 20−50 nm particles were primarily determined by their surface charges and ability to penetrate the cellular membrane, and the final destinations of the nanoparticles in the cell could be controlled by the appropriate design of surface ligands.

Sedentary behaviors, like television viewing, are positively associated with overweight among young people. To monitor national health objectives for sedentary behaviors in young adolescents, this project developed and assessed the reliability and validity of a brief questionnaire to measure weekly television viewing, usual television viewing, and computer use by middle school children. Reliability and validity of the Youth Risk Behavior Survey (YRBS) question on weekday television viewing also were examined. A brief, five-item television and computer use questionnaire was completed twice by 245 middle school children with one week apart. To concurrently assess validity, students also completed television and computer use logs for seven days. Among all students, Spearman correlations for test-retest reliability for television viewing and computer use ranged from 0.55 to 0.68. Spearman correlations between the first questionnaire and the seven-day log produced the following results: YRBS question for weekday television viewing (0.46), weekend television viewing (0.37), average television viewing over the week (0.47), and computer use (0.39). Methods comparison analysis showed a mean difference (hours/week) between answers to questionnaire items and the log of -0.04 (1.70 standard deviation [SD]) hours for weekday television, -0.21 (2.54 SD) for weekend television, -0.09 (1.75 SD) for average television over the week, and 0.68 (1.26 SD) for computer use. The YRBS weekday television viewing question, and the newly developed questions to assess weekend television viewing, average television viewing, and computer use, produced adequate reliability and validity for surveillance of middle school students.

Renal cells are used in basic research, disease models, tissue engineering, drug screening, and in vitro toxicology. In order to provide a reliable source of human renal cells, we developed a protocol for the differentiation of human embryonic stem cells into renal epithelial cells. The differentiated stem cells expressed markers characteristic of renal proximal tubular cells and their precursors, whereas markers of other renal cell types were not expressed or expressed at low levels. Marker expression patterns of these differentiated stem cells and in vitro cultivated primary human renal proximal tubular cells were comparable. The differentiated stem cells showed morphological and functional characteristics of renal proximal tubular cells, and generated tubular structures in vitro and in vivo. In addition, the differentiated stem cells contributed in organ cultures for the formation of simple epithelia in the kidney cortex. Bioreactor experiments showed that these cells retained their functional characteristics under conditions as applied in bioartificial kidneys. Thus, our results show that human embryonic stem cells can differentiate into renal proximal tubular-like cells. Our approach would provide a source for human renal proximal tubular cells that are not affected by problems associated with immortalized cell lines or primary cells.