7 research outputs found
NāDoped Graphene: An Alternative Carbon-Based Matrix for Highly Efficient Detection of Small Molecules by Negative Ion MALDI-TOF MS
Gas-phase
N-doped graphene (gNG) was synthesized by a modified
thermal annealing method using gaseous melamine as nitrogen source
and then for the first time applied as a matrix in negative ion matrix-assisted
laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF
MS) for small molecule analysis. Unlike the complicated adducts produced
in positive ion mode, MS spectra obtained on gNG matrix in negative
ion mode was only featured by deprotonated molecule ion peaks without
matrix interference. By the gNG assisted desorption/ionization (D/I)
process, some applications were carried out on a wide range of low-molecular
weight (MW) analytes including amino acids, fatty acids, peptides,
anabolic androgenic steroids as well as anticancer drugs, with an
extraordinary laser desorption/ionization (LDI) efficiency over traditional
Ī±-cyano-4-hydroxycinnamic acid (CHCA) and other carbon-based
materials in the negative ion detection mode. By comparison of a series
of graphene-based matrixes, two main factors of matrix gNG were unveiled
to play a decisive role in assisting negative ion D/I process: a well-ordered
Ļ-conjugated system for laser absorption and energy transfer;
pyridinic-doped nitrogen species functioning as deprotonation sites
for proton capture on negative ionization. The good salt tolerance
and high sensitivity allowed further therapeutic monitoring of anticancer
drug nilotinib in the spiked human serum, a real case of biology.
Signal response was definitely obtained between 1 mM and 1 Ī¼M,
meeting the demand of assessing drug level in the patient serum. This
work creates a new application branch for nitrogen-doped graphene
and provides an alternative solution for small molecule analysis
Multiplexed Quantitative MALDI MS Approach for Assessing Activity and Inhibition of Protein Kinases Based on Postenrichment Dephosphorylation of Phosphopeptides by MetalāOrganic Framework-Templated Porous CeO<sub>2</sub>
Protein
kinase is regarded as a potential target for anticancer therapeutics
due to its relation to many diseases, in which more than one kinase
participates in the cell signaling cascades. We herein demonstrate
a multiplexed quantitative matrix-assisted laser desorption/ionization
mass spectrometry (MALDI MS) approach to simultaneously assess the
activity and inhibition of multiple protein kinases. In this design,
substrate peptides phosphorylated by kinases of interest are specifically
harvested by metalāorganic framework (MOF)-templated porous
CeO<sub>2</sub> and consequently transformed to the dephosphorylated
forms due to the phosphatase-like activity of CeO<sub>2</sub>, resulting
in a unique quantitative MS signal with an enhanced intensity. Based
on the peak area ratios of dephosphorylated variants of the phosphorylated
product to respective deuterated internal standard, the activity and
inhibition of each kinase can be independently profiled. In addition
to the accurate characterization of protein kinase A (PKA) activity
and inhibition induced by H-89, the multiplexing capability of the
MS-based method allowed quantitative evaluation of the activity of
Abl and Src, the two tightly associated kinases in the occurrence
of chronic myeloid leukemia (CML) in a multiplexed format, exhibiting
excellent orthogonality for the dual signal readout channels. Moreover,
the inactivation of both Abl and Src by the inhibitor imatinib, dasatinib,
and ponatinib was simultaneously traced, giving a whole picture of
the competition behavior between the kinases for binding therapeutic
molecules. This approach holds great promise in global investigation
of kinase signal pathways and high-throughput screening of effective
protein kinase inhibitors
āStealth and Fully-Ladenā Drug Carriers: Self-Assembled Nanogels Encapsulated with Epigallocatechin Gallate and siRNA for Drug-Resistant Breast Cancer Therapy
For
codelivery of therapeutic genes and chemical agents in combined therapy,
the ideal drug delivery system entails high-capacity and low-body
toxicity carriers, allowing adequate drug dose for tumor regions while
yielding low residues in normal tissues. To augment the gene/drug
load capacity and circumvent the potential toxicity brought by traditional
inorganic and polymeric nanocarriers, a āstealthā carrier
was herein designed in a simple self-assembly of (ā)-epigallocatechin-3-<i>O</i>-gallate (EGCG) and small interfering RNA (siRNA) by recruiting
protamine as a biodegradable medium for the treatment of drug-resistant
triple-negative breast cancer. In the self-assembled nanogel, entrapped
siRNA played a central role in sensitizing the tumor response to EGCG-involved
chemotherapy, and the positively charged protamine served as the assembly
skeleton to fully accommodate gene and drug molecules and minimize
the factors causing side effects. As compared to stand-alone chemotherapy
with EGCG, the multicomponent nanogel revealed a 15-fold increase
in the cytotoxicity to drug-resistant MDA-MB-231 cell line. Moreover,
equipped with hyaluronic acid and tumor-homing cell-penetrating peptide
as the outmost targeting ligands, the siRNA- and EGCG-loaded nanogel
demonstrates superior selectivity and tumor growth inhibition to free
EGCG in xenograft MDA-MB-231 tumor-bearing mice. Meanwhile, thanks
to the acknowledged biosafety of protamine, little toxicity was found
to normal tissues and organs in the animal model. This gene/drug self-assembly
caged in a biodegradable carrier opens up an effective and secure
route for drug-resistant cancer therapy and provides a versatile approach
for codelivery of other genes and drugs for different medical purposes
Enriching Reaction Intermediates in Multishell Structured Copper Catalysts for Boosted Propanol Electrosynthesis from Carbon Monoxide
Fine-tuned
catalysts that alter the diffusion kinetics of reaction
intermediates is of great importance for achieving high-performance
multicarbon (C2+) product generation in carbon monoxide
(CO) reduction. Herein, we conduct a structural design based on Cu2O nanoparticles and present an effective strategy for enhancing
propanol electrosynthesis from CO. The electrochemical characterization,
operando Raman monitoring, and finite-element method simulations reveal
that the multishell structured catalyst can realize the enrichment
of C1 and C2 intermediates by nanoconfinement
space, leading to the possibility of further coupling. Consequently,
the multishell copper catalyst realizes a high Faraday efficiency
of 22.22 Ā± 0.38% toward propanol at the current density of 50
mA cmā2
Single Gold@Silver Nanoprobes for Real-Time Tracing the Entire Autophagy Process at Single-Cell Level
This
article describes a multimodified coreāshell gold@silver
nanoprobe for real-time monitoring the entire autophagy process at
single-cell level. Autophagy is vital for understanding the mechanisms
of human pathologies, developing novel drugs, and exploring approaches
for autophagy controlling. A major challenge for autophagy study lies
in real-time monitoring. One solution might come from real-time detection
of in situ superoxide radicals (O<sub>2</sub><sup>ā¢ā</sup>), because it is the main regulator of autophagy. In this work, our
proposed nanoprobes were etched by O<sub>2</sub><sup>ā¢ā</sup> and gave a notable wavelength change in the plasmon resonance scattering
spectra. Both the experimental and simulated results suggested the
wavelength change rate correlated well with O<sub>2</sub><sup>ā¢ā</sup> level. This response enabled its application in real-time in situ
quantification of O<sub>2</sub><sup>ā¢ā</sup> during
autophagy course. More importantly, with the introduction of ārelay
probeā operation, two types of O<sub>2</sub><sup>ā¢ā</sup>-regulating autophagy processes were successfully traced from the
beginning to the end, and the possible mechanism was also proposed
<i>In Situ</i> Amplification of Intracellular MicroRNA with MNAzyme Nanodevices for Multiplexed Imaging, Logic Operation, and Controlled Drug Release
MicroRNAs (miRNAs), as key regulators in gene expression networks, have participated in many biological processes, including cancer initiation, progression, and metastasis, indicative of potential diagnostic biomarkers and therapeutic targets. To tackle the low abundance of miRNAs in a single cell, we have developed programmable nanodevices with MNAzymes to realize stringent recognition and <i>in situ</i> amplification of intracellular miRNAs for multiplexed detection and controlled drug release. As a proof of concept, miR-21 and miR-145, respectively up- and down-expressed in most tumor tissues, were selected as endogenous cancer indicators and therapy triggers to test the efficacy of the photothermal nanodevices. The sequence programmability and specificity of MNAzyme motifs enabled the fluorescent turn-on probes not only to sensitively profile the distributions of miR-21/miR-145 in cell lysates of HeLa, HL-60, and NIH 3T3 (9632/0, 14147/0, 2047/421 copies per cell, respectively) but also to visualize trace amounts of miRNAs in a single cell, allowing logic operation for graded cancer risk assessment and dynamic monitoring of therapy response by confocal microscopy and flow cytometry. Furthermore, through general molecular design, the MNAzyme motifs could serve as three-dimensional gatekeepers to lock the doxorubicin inside the nanocarriers. The drug nanocarriers were exclusively internalized into the target tumor cells <i>via</i> aptamer-guided recognition and reopened by the endogenous miRNAs, where the drug release rates could be spatial-temporally controlled by the modulation of miRNA expression. Integrated with miRNA profiling techniques, the designed nanodevices can provide general strategy for disease diagnosis, prognosis, and combination treatment with chemotherapy and gene therapy
Near Infrared-Guided Smart Nanocarriers for MicroRNA-Controlled Release of Doxorubicin/siRNA with Intracellular ATP as Fuel
In chemotherapy, it is a great challenge
to recruit endogenous
stimuli instead of external intervention for targeted delivery and
controlled release; microRNAs are the most promising candidates due
to their vital role during tumorigenesis and significant expression
difference. Herein, to amplify the low abundant microRNAs in live
cells, we designed a stimuli-responsive DNA Y-motif for codelivery
of siRNA and Dox, in which the cargo release was achieved <i>via</i> enzyme-free cascade amplification with endogenous microRNA
as trigger and ATP (or H<sup>+</sup>) as fuel through toehold-mediated
strand displacement. Furthermore, to realize controlled release in
tumor cells, smart nanocarriers were constructed with stimuli-responsive
Y-motifs, gold nanorods, and temperature-sensitive polymers, whose
surfaces could be reversibly switched between PEG and RGD states <i>via</i> photothermal conversion. The PEG corona kept the nanocarriers
stealth during blood circulation to protect the Y-motifs against nuclease
digestion and enhance passive accumulation, whereas the exposed RGD
shell under near-infrared (NIR) irradiation at tumor sites facilitated
the specific receptor-mediated endocytosis by tumor cells. Through
modulating NIR laser, microRNA, or ATP expressions, the therapy efficacies
to five different cell lines were finely controlled, presenting NIR-guided
accumulation, massive release, efficient gene silence, and severe
apoptosis in HeLa cells; <i>in vivo</i> study showed that
a low dosage of nanocarriers synergistically inhibited the tumor growth
by silencing gene expression and inducing cell apoptosis under mild
NIR irradiation, though they only brought minimum damage to normal
organs. The combination of nanomaterials, polymers, and DNA nanomachines
provided a promising tool for designing smart nanodevices for disease
therapy