Engineered protein cages with enhanced extracellular drug release for elevated antitumor efficacy.

Human heavy chain ferritin (HFn) protein cage has been explored as a nanocarrier for targeted anticancer drug delivery. Here, we introduced a matrix metalloproteinases (MMPs)-cleavable sequence into the DE loop of HFn, creating an MMP-responsive variant, MR-HFn, for localized and extracellular drug release. The crystal structure of MR-HFn revealed that the addition of the MMPs recognition sequence did not affect the self-assembly of HFn but presented a surface-exposed loop susceptible to MMPs cleavage. Biochemical analysis indicated that this engineered protein cage is responsive to MMPs, enabling the targeted release of encapsulated drugs. To evaluate the therapeutic potential of this engineered protein cage, monosubstituted ß-carboxy phthalocyanine zinc (CPZ), a type of photosensitizer, was loaded inside this protein cage. The prepared CPZ@MR-HFn showed higher uptake and stronger phototoxicity in MMPs overexpressed tumor cells, as well as enhanced penetration into multicellular tumor spheroids compared with its counterpart CPZ@HFn in vitro. In vivo, CPZ@MR-HFn displayed a higher tumor inhibitory rate than CPZ@HFn under illumination. These results indicated that MR-HFn is a promising nanocarrier for anticancer drug delivery and the MMP-responsive strategy here can also be adapted for other stimuli.

Restoration of saturated outputs from microchannel plate photomultiplier tubes in sub-microsecond single-pulse-current mode.

Microchannel plate (MCP) photomultiplier tubes (PMTs) are frequently used in experimental diagnostics, where they are operated in single-pulse current measurement mode. However, considering the significant amplitude fluctuations in the measured signal, the resulting output signal from the MCP-PMT is inevitably distorted by gain saturation. Therefore, understanding the correlation between the MCP-PMT output signal and gain saturation is critical in assessing the extent of output signal distortion and determining the MCP-PMT saturation level. This knowledge allows for a more precise assessment of the input signal's features. In this paper, we present an experimental method for restoring the initial waveform from the saturated MCP-PMT signal. To correct the amplitude-drop caused by gain saturation, our technique involves calibrating the MCP-PMT's relative gain as a function of the accumulated output charge using a square-wave light source. We then applied this approach to restore a ∼500 ns saturated pulse from a double-layer 10 mm diameter MCP-PMT. The restored signal showed a deviation of less than 6% from the reference waveform, which validates the effectiveness of the technique.

Identification of Antithrombotic Natural Products Targeting the Major Substrate Binding Pocket of Protein Disulfide Isomerase.

Protein disulfide isomerase (PDI) is a vital oxidoreductase. Extracellular PDI promotes thrombus formation but does not affect physiological blood hemostasis. Inhibition of extracellular PDI has been demonstrated as a promising strategy for antithrombotic treatment. Herein, we focused on the major substrate binding site, a unique pocket in the PDI b' domain, and identified four natural products binding to PDI by combining virtual screening with tryptophan fluorescence-based assays against a customized natural product library. These hits all directly bound to the PDI-b' domain and inhibited the reductase activity of PDI. Among them, galangin showed the most prominent potency (5.9 µM) against PDI and as a broad-spectrum inhibitor for vascular thiol isomerases. In vivo studies manifested that galangin delayed the time of blood vessel occlusion in an electricity-induced mouse thrombosis model. Molecular docking and dynamics simulation further revealed that the hydroxyl-substituted benzopyrone moiety of galangin deeply inserted into the interface between the PDI-b' substrate-binding pocket and the a' domain. Together, these findings provide a potential antithrombotic drug candidate and demonstrate that the PDI b' domain is a critical domain for inhibitor development. Besides, we also report an innovative high-throughput screening method for the rapid discovery of PDI b' targeted inhibitors.

Applications of thin film plastic scintillator in measurement of soft x rays generated from Z-pinch implosion.

A thin film plastic scintillator detector has been developed for the measurement of radiation power and yield of soft x rays produced from Z-pinch implosion. To enable soft x-ray measurements using plastic scintillators, the detector geometry has been specially designed to minimize visible light and alleviate nonlinear behavior. Energy response has been calibrated, and saturation effects have been explored and described in details. The possibility and limitation of its application to such high-density radiation bursts are analyzed. The detector has been fielded on several meters away in vacuum pipes for hundreds of shots at different Z-pinch facilities, and the measured data in these experiments agreed well with the results from other diagnostics, demonstrating the feasibility and reliability of the detector.

A free-standing thin foil bolometer for measuring soft x-ray fluence.

A free-standing thin foil bolometer for measuring soft x-ray fluence in z-pinch experiments is developed. For the first time, we present the determination of its sensitivity by different methods. The results showed great consistency for the different methods, which confirms the validity of the sensitivity and provides confidence for its application in z-pinch experiments. It should be highlighted that the sensitivity of a free-standing foil bolometer could be calibrated directly using Joule heating without any corrections that will be necessary for a foil bolometer with substrate because of heat loss. The difference of the waveforms between the free-standing foil bolometer and that with substrate is obvious. It reveals that the heat loss to the substrate should be considered for the latter in despite of the short x-ray pulse when the peak value is used to deduce the total deposited energy. The quantitative influence is analyzed through a detailed simulation.

Measurements of high energy photons in Z-pinch experiments on primary test stand.

High energy photons are measured for the first time in wire-array Z-pinch experiments on the Primary Test Stand (PTS) which delivers a current up to 8 MA with a rise time of 70 ns. A special designed detecting system composed of three types of detectors is used to measure the average energy, intensity, and pulse waveform of high energy photons. Results from Pb-TLD (thermoluminescence dosimeter) detector indicate that the average energy is 480 keV (±15%). Pulse shape of high energy photons is measured by the photodiode detector consisted of scintillator coupled with a photodiode, and it is correlated with soft x-ray power by the same timing signal. Intensity is measured by both TLD and the photodiode detector, showing good accordance with each other, and it is 10(10) cm(-2) (±20%) at 2 m in the horizontal direction. Measurement results show that high energy photons are mainly produced in pinch regions due to accelerated electrons. PTS itself also produces high energy photons due to power flow electrons, which is one order smaller in amplitude than those from pinch region.