Synthesis and Synergistic Antimicrobial Efficacy of Covalent Conjugates Composed of Epsilon-Poly-l-lysine and Beta-Lactam Antibiotics.

The increasing severity of problems posed by drug-resistant pathogens has compelled researchers to explore innovative approaches for infection prevention. Among these strategies, conjugation methods stand out for their convenience and high efficacy. In this study, multiple covalent conjugates were synthesized, incorporating the natural antimicrobial peptide epsilon-poly-l-lysine (EPL) and two commonly used ß-lactam antibiotics: penicillin G or ampicillin. Enhanced antimicrobial efficacy against typical Gram-negative pathogens, along with faster kill kinetics compared to combination approaches, was demonstrated by the EPL-Ampicillin covalent conjugates. Their antimicrobial mechanism was also substantiated through SEM and fluorescence tests in this work, confirming the inheritance of membrane-disrupting properties from EPL. Furthermore, the excellent biocompatibility of the raw materials was reserved in the covalent conjugates. This simplified conjugation method holds promise for the development of infection therapeutic drugs and potentially restores the sensitivity of conventional antibiotics to drug-resistant pathogens by introducing membrane-disrupting mechanisms.

Therapeutic Effects of Synthetic Triblock Amphiphilic Short Antimicrobial Peptides on Human Lung Adenocarcinoma.

Because of their unique properties, antimicrobial peptides (AMPs) represent a potential reservoir of novel anticancer therapeutic agents. However, only a few AMPs can kill tumors with high efficiency, and obtaining inexpensive anticancer AMPs with strong activity is still a challenge. In our previous work, a series of original short amphiphilic triblock AMP (KnFmKn) analogues were developed which were demonstrated to exert excellent effects on bacterial infection, both in vitro and in vivo. Herein, the overall objectives were to assess the potent tumoricidal capacities of these analogues against human lung cancer cell line A549 and the underlying mechanism. The results of the CCK-8 assay revealed that the precise modification of the peptides' primary sequences could modulate their tumoricidal potency. In the tumoricidal progress, positive charge and hydrophobicity were the key driving forces. Among these peptides, K4F6K4 displayed the most remarkable tumoricidal activity. Furthermore, the excellent anticancer capacity of K4F6K4 was proven by the live/dead cell staining, colony formation assay, and tumor growth observations on xenografted mice, which indicated that K4F6K4 might be a promising drug candidate for lung cancer, with no significant adverse effects in vitro or in vivo. In addition, the cell apoptosis assay using flow cytometry, the morphology observations using the optical microscope, confocal microscopy using CellMask™ Deep Red staining, and scanning electron microscope suggested that membrane disruption was the primary mechanism of its antitumor action. Through analyzing the structure-activity relationship, it was found that the amount of positive charge required for KnFmKn to exert its optimal tumoricidal effect was more than that needed for the antimicrobial activity, while the optimal proportion of hydrophobicity was less. Our findings suggest that further analysis of the structure-activity relationship of AMPs' primary sequence variations will be beneficial. Hopefully, this work can provide guiding principles in designing peptide-based therapeutics for lung cancer.

Contacts with Macrophages Promote an Aggressive Nanomechanical Phenotype of Circulating Tumor Cells in Prostate Cancer.

Aggressive tumors of epithelial origin shed cells that intravasate and become circulating tumor cells (CTC). The CTCs that are able to survive the stresses encountered in the bloodstream can then seed metastases. We demonstrated previously that CTCs isolated from the blood of prostate cancer patients display specific nanomechanical phenotypes characteristic of cell endurance and invasiveness and patient sensitivity to androgen deprivation therapy. Here we report that patient-isolated CTCs are nanomechanically distinct from cells randomly shed from the tumor, with high adhesion as the most distinguishing biophysical marker. CTCs uniquely coisolated with macrophage-like cells bearing the markers of tumor-associated macrophages (TAM). The presence of these immune cells was indicative of a survival-promoting phenotype of "mechanical fitness" in CTCs based on high softness and high adhesion as determined by atomic force microscopy. Correlations between enumeration of macrophages and mechanical fitness of CTCs were strong in patients before the start of hormonal therapy. Single-cell proteomic analysis and nanomechanical phenotyping of tumor cell-macrophage cocultures revealed that macrophages promoted epithelial-mesenchymal plasticity in prostate cancer cells, manifesting in their mechanical fitness. The resulting softness and adhesiveness of the mechanically fit CTCs confer resistance to shear stress and enable protective cell clustering. These findings suggest that selected tumor cells are coached by TAMs and accompanied by them to acquire intermediate epithelial/mesenchymal status, thereby facilitating survival during the critical early stage leading to metastasis. SIGNIFICANCE: The interaction between macrophages and circulating tumor cells increases the capacity of tumor cells to initiate metastasis and may constitute a new set of blood-based targets for pharmacologic intervention.

Polycaprolactone-Based Mimetic Antimicrobial Peptide Copolymers Vesicles as an Effective Drug-Carrier for Cancer Therapy.

A novel series of amphiphilic mimicking antimicrobial peptide copolymers PCL16-b-Kn can assemble in water to form uniform vesicles. Transmission electron microscopy was used to observe the vesicular structure of the nanoparticles, and dynamic light scattering revealed their uniform size and narrow dispersion. Critical vesiculation concentrations were also tested, revealing that these vesicles can exist at low concentrations. Furthermore, in vitro and intracellular drug release of doxorubicin(DOX)-vesicles were conducted. These vesicles could encapsulate DOX and achieve efficient intracellular drug release. Overall, these copolymer vesicles exhibit potential application value as multifunctional drug-carrier systems with antibacterial capability in cancer therapy.

Polymeric Antimicrobial Food Packaging and Its Applications.

Food corruption and spoilage caused by food-borne pathogens and microorganisms is a serious problem. As a result, the demand for antibacterial drugs in food packaging is growing. In this review, biodegradable and non-biodegradable materials for food packaging are discussed based on their properties. Most importantly, antibacterial agents are essential to inhibit the growth of bacteria in food. To keep food fresh and prolong the shelf life, different kinds of antibacterial agents were used. The composition and application of natural antibacterial agents and synthetic antibacterial agents are discussed. Compared with natural antibacterial agents, synthetic antibacterial agents have the advantages of low cost and high activity, but their toxicity is usually higher than that of natural antibacterial agents. Finally, future development of antimicrobial food packaging is proposed. It is an urgent problem for researchers to design and synthesize antibacterial drugs with high efficiency and low toxicity.

GDNF-expressing macrophages mitigate loss of dopamine neurons and improve Parkinsonian symptoms in MitoPark mice.

Glial cell line-derived neurotrophic factor (GDNF) is the most potent neuroprotective agent tested in cellular and animal models of Parkinson's disease (PD). However, CNS delivery of GDNF is restricted by the blood-brain barrier (BBB). Using total body irradiation as transplant preconditioning, we previously reported that hematopoietic stem cell (HSC) transplantation (HSCT)-based macrophage-mediated gene therapy could deliver GDNF to the brain to prevent degeneration of nigrostriatal dopamine (DA) neurons in an acute murine neurotoxicity model. Here, we validate this therapeutic approach in a chronic progressive PD model - the MitoPark mouse, with head shielding to avoid inducing neuroinflammation and compromising BBB integrity. Bone marrow HSCs were transduced ex vivo with a lentiviral vector expressing macrophage promoter-driven GDNF and transplanted into MitoPark mice exhibiting well developed PD-like impairments. Transgene-expressing macrophages infiltrated the midbrains of MitoPark mice, but not normal littermates, and delivered GDNF locally. Macrophage GDNF delivery markedly improved both motor and non-motor symptoms, and dramatically mitigated the loss of both DA neurons in the substantia nigra and tyrosine hydroxylase-positive axonal terminals in the striatum. Our data support further development of this HSCT-based macrophage-mediated GDNF delivery approach in order to address the unmet need for a disease-modifying therapy for PD.

Multimodal MRI Evaluation of the MitoPark Mouse Model of Parkinson's Disease.

The MitoPark mouse, a relatively new genetic model of Parkinson's disease (PD), has a dopaminergic neuron-specific knock-out that inactivates the mitochondrial transcription factor A (Tfam), a protein essential for mitochondrial DNA expression and maintenance. This study used multimodal MRI to characterize the neuroanatomical correlates of PD-related deficits in MitoPark mice, along with functional behavioral tests. Compared with age-matched wild-type animals, MitoPark mice at 30 weeks showed: i) reduced whole-brain volume and increased ventricular volume, indicative of brain atrophy, ii) reduced transverse relaxation time (T2*) of the substantia nigra and striatum, suggestive of abnormal iron accumulation, iii) reduced apparent diffusion coefficient in the substantia nigra, suggestive of neuronal loss, iv) reduced fractional anisotropy in the corpus callosum and substantia nigra, indicative of white-matter damages, v) cerebral blood flow was not significantly affected, and vi) reduced motor activity in open-field tests, reduced memory in novel object recognition tests, as well as decreased mobility in tail suspension tests, an indication of depression. In sum, MitoPark mice recapitulate changes in many MRI parameters reported in PD patients. Multimodal MRI may prove useful for evaluating neuroanatomical correlates of PD pathophysiology in MitoPark mice, and for longitudinally monitoring disease progression and therapeutic interventions for PD.

Demonstration of microcantilever array with simultaneous readout using an in-plane photonic transduction method.

We demonstrate a microcantilever array with an in-plane photonic transduction method for simultaneous readout of each microcantilever. The array is fabricated on a silicon-on-insulator substrate. Rib waveguides in conjunction with a compact waveguide splitter network comprised of trench-based splitters and trench-based bends route light from a single optical input to each microcantilever on the chip. Light propagates down a rib waveguide integrated into the microcantilever and, at the free end of the microcantilever, crosses a small gap. Light is captured in static asymmetric multimode waveguides that terminate in Y-branches, the outputs of which are imaged onto an InGaAs line scan camera. A differential signal for each microcantilever is simultaneously formed from the two outputs of the corresponding Y-branch. We demonstrate that reasonable signal uniformity is obtained with a scaled differential signal for seven out of nine surviving microcantilevers in an array.

Compact waveguide splitter networks.

We demonstrate compact waveguide splitter networks in siliconon- insulator (SOI) rib waveguides using trench-based splitters (TBSs) and bends (TBBs). Rather than a 90 degrees geometry, we use 105 degrees TBSs to facilitate reliable fabrication of high aspect ratio trenches suitable for 50/50 splitting when filled with SU8. Three dimensional (3D) finite difference time domain (FDTD) simulation is used for splitter and bend design. Measured TBB and TBS optical efficiencies are 84% and 68%, respectively. Compact 105 degrees 1 x 4, 1 x 8, and 1 x 32 trench-based splitter networks (TBSNs) are demonstrated. The measured total optical loss of the 1 x 32 TBSN is 9.15 dB. Its size is only 700 microm x 1600 microm for an output waveguide spacing of 50 microm.

Compact 90 degrees trench-based splitter for silicon-on-insulator rib waveguides.

Compact silicon-on-insulator (SOI) rib waveguide 90 degrees splitters based on narrow, high-aspect ratio (~10:1) trenches are designed and experimentally demonstrated. The splitter area is only 11 mum x 11 mum. Splitter optical performance is investigated as a function of both trench width and refractive index of the trench fill material. We examine three trench fill materials, air (n=1.0), SU8 (n=1.57), and index matching fluid (n=1.733), and find good agreement between experimental measurement and three dimensional (3D) finite difference time domain (FDTD) simulation. A splitting ratio of 49/51 (reflection/transmission) is measured for an index fluid-filled trench 82nm wide.

Compact and low loss silicon-on-insulator rib waveguide 90 degrees bend.

A compact and low loss silicon-on-insulator rib waveguide 90 degrees bend is designed and demonstrated. An interface realized by a trench filled with SU8 at the corner of a waveguide bend effectively reflects incoming light through total internal reflection (TIR). In order to accurately position the SU8-filled trench relative to the waveguide and reduce sidewall roughness of the interface, electron beam lithography (EBL) is employed while inductively coupled plasma reactive ion etching (ICP RIE) is used to achieve a vertical sidewall. The measured loss for TE polarization is 0.32 dB +/- 0.02 dB/bend at a wavelength of 1.55 microm.