DNA double-strand breaks (DSBs) are repaired by a hierarchically regulated network of pathways. Factors influencing the choice of particular repair pathways, however remain poorly characterized. Here ...we develop an Integrated Classification Pipeline (ICP) to decompose and categorize CRISPR/Cas9 generated mutations on genomic target sites in complex multicellular insects. The ICP outputs graphic rank ordered classifications of mutant alleles to visualize discriminating DSB repair fingerprints generated from different target sites and alternative inheritance patterns of CRISPR components. We uncover highly reproducible lineage-specific mutation fingerprints in individual organisms and a developmental progression wherein Microhomology-Mediated End-Joining (MMEJ) or Insertion events predominate during early rapid mitotic cell cycles, switching to distinct subsets of Non-Homologous End-Joining (NHEJ) alleles, and then to Homology-Directed Repair (HDR)-based gene conversion. These repair signatures enable marker-free tracking of specific mutations in dynamic populations, including NHEJ and HDR events within the same samples, for in-depth analysis of diverse gene editing events.
To improve the response rate of immune checkpoint inhibitors such as anti-PD-L1 antibody in immunosuppressive cancers like triple-negative breast cancer (TNBC), induction of immunogenic cell death ...(ICD) at tumor sites can increase the antigenicity and adjuvanticity to activate the immune microenvironment so that tumors become sensitive to the intervention of immune checkpoint inhibitors. Herein, a self-amplified biomimetic nanosystem, mEHGZ, was constructed by encapsulation of epirubicin (EPI), glucose oxidase (Gox) and hemin in ZIF-8 nanoparticles and coating of the nanoparticles with calreticulin (CRT) over-expressed tumor cell membrane. EPI acts as an ICD inducer, Gox and hemin medicate the cascade generation of reactive oxygen species (ROS) to strengthen the ICD effect, and CRT-rich membrane as “eat me” signal promote presentation of the released antigens by dendritic cells (DCs) to invoke the tumor-immunity cycle. The biomimetic delivery system displays an amplified ICD effect via Gox oxidation, hydroxyl radical generation and glutathione (GSH) depletion. The induced potent ICD effect promotes DCs maturation and cytotoxic T lymphocytes (CTLs) infiltration, reversing an immunosuppressive tumor microenvironment to an immunoresponsive one. Treatment with the nanosystem in combination with anti-PD-L1 antibody results in distinctive inhibition of tumor growth and lung metastasis, supporting that a potent ICD effect can significantly boost the therapeutic efficacy of the anti-PD-L1 antibody. This self-amplified biomimetic nanoplatform offers a promising means of raising the response rate of immune checkpoint inhibitors.
A biomimetic nanosystem encapsulates epirubicin (EPI), glucose oxidase (Gox) and hemin in the core of ZIF-8 nanoparticles and coats with calreticulin (CRT) over-expressed tumor cell membranes as the outermost layer (mEHGZ), which induces cascade-amplified ICD effect to promote dendritic cells (DCs) maturation and cytotoxic T lymphocytes (CTLs) infiltration into tumor site. These create an immunoactivated tumor microenvironment to boost anti-PD-L1 antibody therapeutic effect. Display omitted
•To improve the response rate of immune checkpoint inhibitor in triple-negative breast cancer, a self-amplified biomimetic nanosystem was prepared.•The nanosystem displayed an amplified immunogenic cell death (ICD) effect via hydroxyl radical generation and glutathione depletion.•The potent ICD effect has promoted dendritic cells maturation and cytotoxic T lymphocytes infiltration into tumor sites to boost the anti-PD-L1 antibody therapeutic effect.
Abstract.
Aiming at the problems of early weak fault feature extraction of bearings in rotating machinery, an improved stochastic resonance (SR) is proposed combined with the advantage of SR to enhance weak ...characteristic signals with noise energy. Firstly, according to the characteristics of the large parameters of the actual fault signal, the amplitude transform coefficient and frequency transform coefficient are introduced to convert the large parameter signal into small parameter signal which can be processed by SR, and the relationship of second-order parameters are introduced. Secondly, a comprehensive evaluation index (CEI) consisted of power spectrum kurtosis, correlation coefficient, structural similarity, root mean square error, and approximate entropy, is constructed through BP neural network. Moreover, this CEI is adopted as fitness function to search the optimal damping coefficient and amplitude transform coefficient with adaptive weight particle swarm optimization (PSO). Finally, according to the improved optimal SR system, the weak fault feature can be extracted. The simulation and experiment verify the effectiveness of the proposed method compared with traditional second-order general scale transform adaptive SR.
The HD-Zip family has a diversity of functions during plant development. In this study, we identify 33 HD-Zip transcription factors in grape and detect their expressions in ovules and somatic ...embryos, as well as in various vegetative organs.
A genome-wide survey for HD-Zip transcription factors in Vitis was conducted based on the 12 X grape genome (V. vinifera L.). A total of 33 members were identified and classified into four subfamilies (I-IV) based on phylogeny analysis with Arabidopsis, rice and maize. VvHDZs in the same subfamily have similar protein motifs and intron/exon structures. An evaluation of duplication events suggests several HD-Zip genes arose before the divergence of the grape and Arabidopsis lineages. The 33 members of HD-Zip were differentially expressed in ovules of the stenospermic grape, Thompson Seedless and of the seeded grape, Pinot noir. Most have higher expressions during ovule abortion in Thompson Seedless. In addition, transcripts of the HD-Zip family were also detected in somatic embryogenesis of Thompson Seedless and in different vegetative organs of Thompson Seedless at varying levels. Additionally, VvHDZ28 is located in the nucleus and had transcriptional activity consistent with the typical features of the HD-Zip family. Our results provide a foundation for future grape HD-Zip gene function research.
The identification and expression profiles of the HD-Zip transcription factors in grape, reveal their diverse roles during ovule abortion and organ development. Our results lay a foundation for functional analysis of grape HDZ genes.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
A stimuli‐responsive polymeric prodrug‐based nanotheranostic system with imaging agents (cyanine5.5 and gadolinium‐chelates) and a therapeutic agent paclitaxel (PTX) is prepared via polymerization ...and conjugating chemistry. The branched polymeric PTX‐Gd‐based nanoparticles (BP‐PTX‐Gd NPs) demonstrate excellent biocompatibility, and high stability under physiological conditions, but they stimuli‐responsively degrade and release PTX rapidly in a tumor microenvironment. The in vitro behavior of NPs labeled with fluorescent dyes is effectively monitored, and the NPs display high cytotoxicity to 4T1 cells similar to free PTX by impairing the function of microtubules, downregulating anti‐apoptotic protein Bcl‐2, and upregulating the expression of Bax, cleaved caspase‐3, cleaved caspase‐9, cleaved‐PARP, and p53 proteins. Great improvement in magnetic resonance imaging (MRI) is demonstrated by these NPs, and MRI accurately maps the temporal change profile of the tumor volume after injection of NPs and the tumor treatment process is also closely correlated with the T1 values measured from MRI, demonstrating the capability of providing real‐time feedback to the chemotherapeutic treatment effectiveness. The imaging‐guided chemotherapy to the 4T1 tumor in the mice model achieves an excellent anti‐tumor effect. This stimuli‐responsive polymeric nano‐agent opens a new door for efficient breast cancer treatment under the guidance of fluorescence/MRI.
This study demonstrates a strategy to fabricate stimuli‐responsive branched polymeric prodrug‐based theranostic nanomedicine. The anticancer mechanism is studied well and the in vitro and vivo behaviors including biodistribution, retention, and anticancer efficacy can be monitored well by imaging. This stimuli‐responsive polymeric demonstrates great potential of a platform for cancer nanotheranostics.
Linear and cross-linked amphiphilic block polymer-gadolinium conjugates were prepared, and their self-assembled systems with spherical or worm-like morphology were comparatively investigated as ...efficient and safe MR contrast agents.
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•Two-step RAFT polymerization and cross-linking to fabricate amphiphilic polymers.•Modulating the self-assembled morphology by altering the cross-linking degree.•A higher r1 value and better tumor contrast enhancement from the spherical agent.•Sharper MR images of terminal branched vessels in the liver from the worm-like agent.
Although polymer-gadolinium conjugates-based contrast agents (CAs) have shown great promise in safe and efficient magnetic resonance imaging (MRI), very few studies on the use of self-assembled polymers to prepare CAs have been reported. The effect of the self-assembled architectural structures of CAs on their MRI performance remains unclear. Herein, two block amphiphilic polymer-gadolinium conjugates (Linear Agent and Cross-Linked Agent3) were designed and fabricated via an approach of two-step reversible addition-fragmentation chain transfer (RAFT) polymerization and cross-linking. These conjugates self-assembled into nanoscale CAs with a spherical or worm-like morphology. These CAs had low toxicity and great hemocompatibility. Both of them displayed an evident increase in the longitudinal relaxivity and the blood circulation time over small molecular CAs (i.e., Gd-DTPA). Linear Agent exhibited superior MRI signal enhancement to Cross-Linked Agent3 in both subcutaneous and orthotopic breast tumor sites. Cross-Linked Agent3 was easily uptaken by tumor cells, but its tumor-specific responsive degradation contributed to a decrease in its MRI efficiency. In the major organs, two agents were able to generate clear and long-lasting images. Moreover, their long residence time in the blood endowed them as magnetic resonance angiography (MRA) agents. A large number of blood vessels, including liver vessels and kidney vessels, could be clearly observed after injection of both CAs, and the contrast-enhanced MR signals in these vessels lasted for over 4 h. Cross-Linked Agent3 excelled over Linear Agent in displaying the terminal of branch blood vessels in the liver and kidney. Overall, the strategy to prepare self-assembled CAs with tunable morphologies would allow a better understanding of the structure–function relationship of these CAs and provide insights into novel designs of polymer-gadolinium conjugates.
In order to diagnose an incipient fault in rotating machinery under complicated conditions, a fast sparse decomposition based on the Teager energy operator (TEO) is proposed in this paper. In this ...proposed method, firstly, the TEO is employed to enhance the envelope of the impulses, which is more sensitive to frequency and can eliminate the low-frequency harmonic component and noise; secondly, a smoothing filtering algorithm was adopted to suppress the noise in the TEO envelope; thirdly, the fault signal was reconstructed by multiplication of the filtered TEO envelope and the original fault signal; finally, sparse decomposition was used based on a generalized S-transform (GST) to obtain the sparse representation of the signal. The proposed preprocessing method using the filtered TEO can overcome the interference of high-frequency noise while maintaining the structure of fault impulses, which helps the processed signal perform better on sparse decomposition; sparse decomposition based on GST was used to represent the fault signal more quickly and more accurately. Simulation and application prove that the proposed method has good accuracy and efficiency, especially in conditions of very low SNR, such as impulses with anSNR of −8.75 dB that are submerged by noise of the same amplitude.
The core components of CRISPR-based gene drives, Cas9 and guide RNA (gRNA), either can be linked within a self-contained single cassette (full gene-drive, fGD) or be provided in two separate elements ...(split gene-drive, sGD), the latter offering greater control options. We previously engineered split systems that could be converted genetically into autonomous full drives. Here, we examine such dual systems inserted at the spo11 locus that are recoded to restore gene function and thus organismic fertility. Despite minimal differences in transmission efficiency of the sGD or fGD drive elements in single generation crosses, the reconstituted spo11 fGD cassette surprisingly exhibits slower initial drive kinetics than the unlinked sGD element in multigenerational cage studies, but then eventually catches up to achieve a similar level of final introduction. These unexpected kinetic behaviors most likely reflect differing transient fitness costs associated with individuals co-inheriting Cas9 and gRNA transgenes during the drive process.
Multi-modal therapeutics are emerging for simultaneous diagnosis and treatment of cancer. Polymeric carriers are often employed for loading multiple drugs due to their versatility and controlled ...release of these drugs in response to a tumor specific microenvironment. A theranostic nanomedicine was designed and prepared by complexing a small gadolinium chelate, conjugating a chemotherapeutic drug PTX through a cathepsin B-responsive linker and covalently bonding a fluorescent probe pheophorbide a (Ppa) with a branched glycopolymer. The branched prodrug-based nanosystem was degradable in the tumor microenvironment with overexpressed cathepsin B, and PTX was simultaneously released to exert its therapeutic effect. The theranostic nanomedicine, branched glycopolymer-PTX-DOTA-Gd, had an extended circulation time, enhanced accumulation in tumors, and excellent biocompatibility with significantly reduced gadolinium ion (Gd3+) retention after 96 h post-injection. Enhanced imaging contrast up to 24 h post-injection and excellent antitumor efficacy with a tumor inhibition rate more than 90% were achieved from glycopolymer-PTX-DOTA-Gd without obvious systematic toxicity. This branched polymeric prodrug-based nanomedicine is very promising for safe and effective diagnosis and treatment of cancer.
•A cathepsin B-responsive theranostic nanomedicine (glycopolymer-PTX-DOTA-Gd) based on a branched glycopolymer was prepared.•Glycopolymer-PTX-DOTA-Gd can be specifically degradated and release drug at tumor enviornment.•Glycopolymer-PTX-DOTA-Gd enhance the contrast of magnetic resonance imaging (MRI) at tumor sites.•The nanomedicine have good biocompatibility, excellent tumor targeting and anti-tumor efficacy.
Cathepsin B-responsive biodegradable branched glycopolymer-based prodrug was designed and prepared as theranostic nanomedicine for cancer treatments, resulting in significantly enhanced therapeutic indexes and strengthened contrast of magnetic resonance imaging (MRI) at tumor sites. Display omitted
Evaporation ducts are abnormal states of the atmosphere in the air–sea boundary layer that directly affect the propagation trajectory of electromagnetic (EM) waves. Therefore, an accurate diagnosis ...of the evaporation duct height (EDH) is important for studying the propagation trajectory of EM waves in evaporation ducts. Most evaporation duct models (EDMs) based on the Monin–Obukhov similarity theory are empirical methods. Different EDMs have different levels of environmental adaptability. Evaporation duct diagnosis methods based on machine learning methods only consider the mathematical relationship between data and do not explore the physical mechanism of evaporation ducts. To solve the above problems, this study observed the meteorological and hydrological parameters of the five layers of the low-altitude atmosphere in the East China Sea on board the research vessel Xiangyanghong 18 in April 2021 and obtained the atmospheric refractivity profile. An evaporation duct multimodel fusion diagnosis method (MMF) based on a library for support vector machines (LIBSVM) is proposed. First, based on the observed meteorological and hydrological data, the differences between the EDH diagnosis results of different EDMs and MMF were analyzed. When ASTD ≥ 0, the average errors of the diagnostic results of BYC, NPS, NWA, NRL, LKB, and MMF are 2.57 m, 2.92 m, 2.67 m, 3.27 m, 2.57 m, and 0.24 m, respectively. When ASTD < 0, the average errors are 2.95 m, 2.94 m, 2.98 m, 2.99 m, 2.97 m, and 0.41 m, respectively. Then, the EM wave path loss accuracy analysis was performed on the EDH diagnosis results of the NPS model and the MMF. When ASTD ≥ 0, the average path loss errors of the NPS model and MMF are 5.44 dB and 2.74 dB, respectively. When ASTD < 0, the average errors are 5.21 dB and 3.46 dB, respectively. The results show that the MMF is suitable for EDH diagnosis, and the diagnosis accuracy is higher than other models.
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