The emergence of human severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses significant challenges to global public health. Despite the extensive efforts of researchers worldwide, there remains considerable opportunities for improvement in timely diagnosis, specific treatment, and effective vaccines for SARS-CoV-2. This is due, in part, to the large number of asymptomatic carriers, rapid virus mutations, inconsistent confinement policies, untimely diagnosis and limited clear treatment plans. The emerging of nanozymes offers a promising approach for combating SARS-CoV-2 due to their stable physicochemical properties and high surface areas, which enable easier and multiple nano-bio interactions in vivo. Nanozymes inspire the development of sensitive and economic nanosensors for rapid detection, facilitate the development of specific medicines with minimal side effects for targeted therapy, trigger defensive mechanisms in the form of vaccines, and eliminate SARS-CoV-2 in the environment for prevention. In this review, we briefly present the limitations of existing countermeasures against coronavirus disease 2019 (COVID-19). We then reviewed the applications of nanozyme-based platforms in the fields of diagnostics, therapeutics and the prevention in COVID-19. Finally, we propose opportunities and challenges for the further development of nanozyme-based platforms for COVID-19. We expect that our review will provide valuable insights into the new emerging and re-emerging infectious pandemic from the perspective of nanozymes. Nanozyme-based platforms have demonstrated improved sensitivity for SARS-CoV-2 antigen detection, reduced cost, and facilitated rapid diagnosis, making them a promising tool for COVID-19 diagnostics.Nanozyme-based platforms offer potential benefits in targeted therapy for COVID-19 due to their ability to specifically counteract and eliminate SARS-CoV-2 with potentially fewer side effects.Nanozyme-based platforms have been shown to act as immunostimulatory molecules, activating the defense response of the immune system against SARS-CoV-2, potentially providing a therapeutic avenue for COVID-19 treatment. As of May 22, 2023, over 765 million individuals have been affected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), resulting in over six million confirmed deaths. SARS-CoV-2 is extremely infectious and can be transmitted between people within six feet . At the Enzymes, with significant catalytic activity, control the basic metabolic and life-sustaining activities of biological systems on the earth . Most na. utilized Fe3O4 magnetic nanoparticles (MNPs) as a nanozyme probe to detect Ebola virus [. targeted the lipid envelope of influenza virus and destroyed the integrity of neighboring proteins by using iron oxide nanozymes (IONzymes), causing its inactivation [In the past few decades, nanozymes have shown their great effectiveness in fighting against various viruses. Duan et alla virus . The sentivation . Inspiretivation . 1) Nan Nan. utiIn this review, we discussed the limitations of existing countermeasures in combating COVID-19. Moreover, we reviewed the applications of nanozyme-based platforms in the fields of diagnosis, treatment and prevention of COVID-19 and their safety considerations. Finally, we proposed opportunities and challenges for the further development of nanozyme-based platforms for COVID-19. We expect that our paper will put forward new opinions in facing the COVID-19 pandemic from the perspective of nanozymes  method in nucleic acid detection is the gold standard for early diagnosis of COVID-19 . Viral RAlthough nucleic acid detection is greatly encouraged by the WHO, it requires biosafety laboratories, professional instruments and skilled experts . MoreoveRegarding antibody detection, the results may be false-positive due to cross-reactivity when the antibody is bound to an antigen that is distinct from the targeted antigen . The reaCompared with nucleic acid testing or antibody testing, antigen detection can detect the virus itself, which is convenient for large-scale population screening . HoweverIt is necessary to control the incidence of COVID-19 by quick diagnosis of emerging patients . There iNanozymes with high surface volume ratios enable them to have large reservoirs and abundant anchoring sites to load and deliver agents, so they can diagnose various virus infections quickly, sensitively and in real time . It may . investigated a novel chemiluminescence paper using nanozymes for rapid and highly responsive determination of SARS-CoV-2 spike antigen [Traditional antigen detection is mainly based on colloidal gold immune assays or fluorometric immunochromatographic assays . Liu et  antigen   protein of SARS-CoV-2 [Colorimetric assays have potential applications in detecting SARS-CoV-2. Bartolomeo et al. utilized gold nanoparticles (NPs) to cause a change in the extinction spectra of the relevant solutions within just a few minutes to obtain a rapid and credible determination of SARS-CoV-2 . In addiRS-CoV-2 . They pr. built a nanozyme-mediated colorimetric detection method for SARS-CoV-2 using a CRISPR‒Cas12a-based system [2 nanorods and label magnetic beads in the presence of SARS-CoV-2, which reduces the oxygenation of tetramethylbenzidine, triggering color variation. The application of MnO2 nanozymes in place of naturally occurring enzymes allows the usage of H2O2 to be avoided, making it cost-effective and simple to operate. In addition, chromatographic analyses based on nanozymes and CRISPR‒Cas12a systems allow the measurement to be easily manipulated and are suitable for high volume microtiter-based analyses. The MnO2 nanozyme-guided CRISPR‒Cas12a system could be used as an outstanding tool for testing SARS-CoV-2.Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) based testing systems have been used to detect nucleic acids –52, protd system   for the quantification of SARS-CoV-2 nucleocapsid protein antigens in human blood samples [Liang et al samples . Au@Pt NThe following categories of medicines are being investigated or are in development for the control of COVID-19: antiviral medicines (such as remdesivir and favepiravir), anti-inflammatories (dexamethasone and statins), antibodies , targeted immunomodulatory therapies , anticoagulants (such as heparin), and antifibrotic medicines (such as tyrosine kinase inhibitors) . Pfizer Virus inhibition is the most effective measure in the earlier stage of the infection. Hydroxychloroquine is hypothesized to suppress viral entrance and endocytosis in vitro and may produce favorable effects of immunomodulation in vivo . LopinavIn the context of the quick spread of SARS-CoV-2 and the growing demand for a rapid response, a globally recognized specific treatment is still lacking. The main goal of current medicines is to control excessive inflammatory and immune reactions in serious conditions . Most of°C. Multiwall and single-wall carbon nanotubes, carbon dots and carbon nanodiamonds both have the potential to directly suppress virus reproduction [As with most nanomaterials, nanozymes are also used as treatment options for diseases, such as serving as drug carriers or passively targeting diverse antiviral treatments . Variousoduction . Due to The structural proteins of SARS-CoV-2 are composed of nucleocapsid protein (N), membrane protein (M), envelope protein (E), and spike protein (S). . When SADue to reasons related to existing at the nanoscale, most nanozymes may have high affinity for virus attachment. Strong nano-bio interactions can recognize virus structures in situ. Nanozymes can interact with viral structures and directly inactivate or destroy them without harming host cells, which will minimize their side effects and degradation , 82. Com2 single-atom nanozymes (SANs) with significant inhibition of SARS-CoV-2 entry activities  nanozymes to imitate natural glutathione peroxidase activity in function, catalyzing ROS neutralization in virus-infected cells to mitigate ROS and uniformly block viral reactivation and replication without adverse effects on cell physiology [V+2O5 nanosheets against SARS-CoV-2 and suggested nanozymes as future platforms to develop interventions against COVID-19.Viral infection and replication are closely associated with oxidative stress. Singh et alysiology . Meanwhi2 nanocatalyst and Ce6 photosensitizer and biomimetic shells of macrophage membrane. The intense expression of cell membrane receptors in MCeC@MΦ allows the binding of decoy nanozymes to endotoxins or proinflammatory cytokines, which neutralizes endotoxin, sequesters proinflammatory cytokines and provides protection against overactivation of host immune cells [Escherichia coli bacteremia mice. This might open the possibilities of treating cytokine storms and immune-mediated inflammatory diseases caused by COVID-19.During infection, excessive immune responses may cause overwhelming immune system impairment in the human body –88. Highne cells . At the Epidemiological data show that SARS-CoV-2 is mainly transmitted through airway splashing during intimate face-to-face exposure, such as talking, coughing or sneezing . Other pNumerical modeling research and empirical data suggest that increasing the strength of public health care intervention can reduce the transmission rate of COVID-19 around the world –98. PublWith the COVID-19 pandemic, many countries have put forward the concept of “herd immunity” . Herd imEffective vaccination is the best way to accelerate herd immunity and eliminate epidemics without causing more deaths . Recentl. developed a chitosan (CS)-functionalized iron oxide nanozyme (IONzyme)-based influenza whole inactivated virus (WIV) nasal vaccine [Nanozymes provide an ideal antigen delivery platform for vaccines, acting as adjacent platforms and mimicking virus structures, and they allow multivalent antigen presentation and antigen stability after administration . Nanozym vaccine . CS‐IONz. sprayed a mixture of copper and shellac nanoparticles onto a nonwoven medical surgical mask, conferring self-sterilizing and reusability ability to the masks [.Kumar et alhe masks . CuNPs e2 at a wavelength of 375 nm). Mechanistic research has shown that light-induced hydroxyl radicals target the viral genome, resulting in viral disintegration, instead of attacking the virion global structure and content [SARS-CoV-2 particles can remain on surfaces for a long time, which provide another route of transmission: surface contact . Nanozym content . Similar content . Due to Despite nanozymes having numerous benefits for combating SARS-CoV-2, including high physical and chemical stability, excellent durability and simplicity, many challenges still remain.One of the main challenges is the lack of knowledge about the underlying mechanisms of nanozymes drugs action with COVID-19. Additional studies are required to fully capture the biological and biochemical interactions between nanozymes and SARS-CoV-2 as well as the potential side effects and toxicity of these biocatalysts. Once nanozymes enter the human body, they can interact with biological molecules (mainly proteins), giving nanozymes new biological characteristics, which may be more significant than the original properties of nanozymes .Another challenge is the lack of standardization in the design and production of nanozymes, which can lead to varying levels of efficacy and specificity. For instance, Au nanozymes are poisonous at 1.4 nm but not at 15 nm. It is crucial to closely explore the characteristic parameters of nanodrugs, particularly the relations among dimension, surface chemistry, dose and pharmacokinetics. Their physicochemical characteristics, such as dimension, geometry, electrical charge or surface chemistry, can be modified if necessary. Modulating the chemical composition and controlling the morphology to change the size, surface modification and assembly of nanozymes provide a reference for researchers in making nanozymes with maximum security for practical biomedical applications.SARS-CoV-2 is constantly mutating , and nanTo hasten the clinical adoption of nanozyme-based vaccines, a comprehensive assessment of their benefits and risks is essential. Safety is one of the most important concerns. Controlled dosage and surface modification can help to reduce toxicity and increase specificity for SARS-CoV-2, but their relevant influence on the following dynamics and catalytic activity of nanozymes must be evaluated. The immunogenicity and cellular transmigration of nanozymes must be researched. Regulation of reaction conditions at larger scales is challenging when producing nanozyme-based vaccines for SARS-CoV-2, so further expansion synthesis must be validated. Otherwise, issues such as batch variation will inevitably hinder the vaccines’ industrialization process.In the context of the SARS-CoV-2 global health outbreak, we have concluded the possible applications of nanozymes in fighting against SARS-CoV-2. (1) Regarding diagnosis, nanozyme-based detection provides a highly sensitive, rapid, inexpensive and even quantitative detection method compared to the current gold standard—the nucleic acid test. (2) In terms of treatment, they can specifically resist and clear SARS-CoV-2 by inhibiting its endocytosis or replication in the process of infecting host cells, which make up for the shortcomings of broad-spectrum medicines. Moreover, direct destruction of the viral structure can minimize its side effects and degradation. (3) Regarding prevention, nanozyme-based vaccines have better potential for appropriate exposure to the immune system and can protect vaccine antigens from degradation and removal. Nanozyme-based masks and surface coating will greatly reduce SARS-CoV-2 in the environment, reducing economic and environmental costs.Although considerable progress has been made in recent years, there is still space for progress in the study of nanozymes in the fight against COVID-19. Here, we propose several key questions and directions of studies that can be researched in the future to increase the potential of clinical transformation of nanozymes. (1) For detection, to commercialize the nanozyme-mediated detection strategy, it is still necessary to evaluate nanozyme-based tests and other commercial kits. (2) For nanozyme-based medicines, we should evaluate their safety, pharmacokinetics, clinical toxicity, immunogenicity, and cellular fate. We should also consider the metabolism and subsequent catalytic activity of nanozymes in vivo. (3) For nanozyme-based vaccines, mass production and availability to the entire range of countries are the target directions. In conclusion, nanozyme-based tools are expected to play a frontline role in addressing COVID-19. Considering the continuous evolution of SARS-CoV-2, nanozymes must also "evolve" in morphology, composition and surface function to counteract.


Depicting Lagged Causality), a convolutional neural network for the inference of gene-regulatory relationships across pseudotime-ordered single-cell trajectories. We show that combining supervised deep learning with joint probability matrices of pseudotime-lagged trajectories allows the network to overcome important limitations of ordinary Granger causality-based methods, for example, the inability to infer cyclic relationships such as feedback loops. Our network outperforms several common methods for inferring gene regulation and, when given partial ground-truth labels, predicts novel regulatory networks from single-cell RNA sequencing (scRNA-seq) and single-cell ATAC sequencing (scATAC-seq) data sets. To validate this approach, we used DELAY to identify important genes and modules in the regulatory network of auditory hair cells, as well as likely DNA-binding partners for two hair cell cofactors (Hist1h1c and Ccnd1) and a novel binding sequence for the hair cell-specific transcription factor Fiz1. We provide an easy-to-use implementation of DELAY under an open-source license at https://github.com/calebclayreagor/DELAY.Identifying the causal interactions in gene-regulatory networks requires an accurate understanding of the time-lagged relationships between transcription factors and their target genes. Here we describe DELAY (short for  The sequencing of genes expressed in single cells provides detailed information about the developmental programs that define the identities and states of various cell types, but few computational methods can use the dynamic information encoded in these representations to identify causal mechanisms. By exploiting advances in machine learning, we develop a deep neural network that learns from temporal features of gene regulation to identify direct regulatory interactions between transcription factors and their target genes. Our method provides mechanistic insights into the development and function of cells by generating high-confidence predictions for interactions in complex gene-regulatory networks.Single-cell sequencing technologies can provide detailed data for the investigation of heterogeneous populations of cells collected at specific times—so-called snapshots—during cellular differentiation or dynamic responses to stimulation . HoweverSeveral methods for gene-regulatory inference rely on pseudotime in Granger causality tests, which try to determine whether new time series can add predictive power to inferred models of gene regulation , 9. HoweDepicting Lagged Causality (DELAY) that learns gene-regulatory interactions from discrete joint probability matrices of paired, pseudotime-lagged gene expression trajectories. Our data suggest that DELAY can address many shortcomings of current Granger causality-based methods and provide a useful, complementary approach to overcome common limitations in the inference of gene-regulatory networks from single-cell data.By contrast, deep learning-based methods make no assumptions about the temporal relationships or connectivity between genes in complex regulatory networks; instead, these data-driven approaches learn general features of regulatory interactions , 16. HerA at several lagged positions in pseudotime relative to a target gene B to generate joint probability distributions from two-dimensional histograms of gene coexpression at each lag  data , we condity test , in whicWe trained DELAY on gene expression data sets from human embryonic stem cells (hESCs) , mouse eAfter training DELAY on the combined 70% splits, the network outperformed eight of the most popular approaches for inferring gene-regulatory relationships, including six unsupervised methods , 22–26  To test whether DELAY generalizes to new data sets, we examined the human hepatocyte (hHep) gene-regulatory network using an additional data set with over 400 single cells and known ground-truth interactions from ChIP-seq data , 28. We Using different numbers of pseudotime-lagged gene coexpression matrices or neighbor gene matrices, as well as examples from modified data sets with fewer cells or additional gene dropouts, we next examined the performance of DELAY across various input configurations. We employed the original data sets to train new models on gene coexpression matrices of up to 10 pseudotime lags Fig. A or up tWe also characterized DELAY's performance on pseudotime-shuffled trajectories and observed a sharp decrease in performance after reordering nearby cells in each trajectory Fig. B. This rces Fig. G. By perces Fig. .C with known interactions across several classes of three-gene motifs. First, we used DELAY to infer gene regulation for potential hierarchical and cyclic interactions across validation set examples  terms across correctly inferred transcription factors Fig. B. We fouWe next sought to generate predictions for a cell type with complex but incompletely characterized gene-regulatory dynamics. We devised a pipeline for multi-omic transfer learning to infer the gene-regulatory network for developing mouse outer hair cells, the mechanical amplifiers of the inner ear. We used both gene expression  and chroTo determine whether pseudotime-lagged gene coaccessibility matrices contain features that indicate direct gene-regulatory relationships, we fine-tuned DELAY on the scATAC-seq matrices using ground-truth examples collected from two cell type–specific data sets of Sox2 and Atoh1 target genes , 33. SevWe separately fine-tuned the network on all available Sox2 and Atoh1 ground-truth examples from an additional gene expression data set for mouse outer hair cells . With prSox2, Id2, Hes1, and Prox1 and hair cell-specific genes such as Atoh1, Pou4f3, Gfi1, Lhx3, and Barhl1. We sought to validate the predicted interactions by comparing the locations of known transcription factor-binding sites . This result implies a role for Lhx3 in maintaining the later expression of these important early hibitors , 47, 49.n humans .cis-regulatory sequences to identify and discard false-positive interactions resulting from indirect gene regulation . Through this analysis, we were able to determine that the most significantly enriched sequence in the enhancers of the histone Hist1h1c's predicted target genes closely matched known binding motifs for several SoxB2/A transcription factors Fig. B. In addSox2 and Hes6. Discriminative motif analysis of these genes’ enhancer sequences , and target–neighbor (for two neighbors). We concatenated the input channels for each example to form three-dimensional stacks of input matrices with dimensions We later used the original randomly segregated validation splits to characterize our network on augmented coexpression matrices and matrices generated from modified single-cell data sets. To augment gene coexpression matrices, we masked with zeros either specific region across all input channels or entire input channels corresponding to specific pseudotime lags.i to find the features for a given output channel j, as shown in Equation We designed a convolutional neural network based on an inverted VGGnet  that useN is the mini-batch size, C is the number of channels, in which the weights and bias are trainable parameters, As shown in Equation x represents the j. After the final convolutional layer, we used global-average pooling  to optimize our network with respect to the class-weighted binary cross-entropy loss, summed across each mini-batch and scaled by the overall batch size 512, as shown in Equation y and N is the size of the current mini-batch (which might be <512). Prior to optimization, we used He initialization  to analyze GO-term enrichment across correctly inferred transcription factors for terms related to InterPro DNA-binding domains SnapATAC  m CIS-BP , or to am CIS-BP , UniPROBm CIS-BP  (mouse),m CIS-BP  databasem CIS-BP  (v5.4.1)pgad113_Supplementary_DataClick here for additional data file.


JCI, Kawai et al. present an adjuvant-free subunit (RBD-HA) fusion vaccine, which produces robust IgG and IgA antibody responses against SARS-CoV-2, enriched within the nasal cavity, by using the host’s preexisting immunity to influenza infection. This preclinical study has tremendous implications for future mucosal vaccine design and provides a roadmap for generating a safer and effective intranasal vaccine against pulmonary infections.Despite the worldwide application of vaccination and other antiviral interventions, pulmonary viral infections remain a persistent threat to human health. The 1918 influenza pandemic killed more than 40 million people in just one year, and the SARS-CoV-2 pandemic has killed more than 6.9 million people since 2019. While the current approved COVID-19 vaccines are administered parenterally and induce systemic immunity, they only prevent the progression to severe disease. Thus, other vaccine platforms are still needed for completely preventing the disease and subsequent transmission. In this issue of the  JCI, Kawai and coauthors report on their generation of an adjuvant-free vaccine containing the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein fused with the hemagglutinin (HA) glycoprotein derived from influenza A virus (IAV)  . The autus (IAV)   .The authors employed several vaccination methods to test the efficacy of their fusion vaccine: RBD-HA administered i.n., RBD plus the adjuvant c-di-GMP administered i.n., and RBD plus the adjuvant alum administered s.c. Following 30 and 51 days of IAV infection, the mice were immunized with a prime/boost vaccine strategy, respectively. RBD-specific antibodies were only detected in mice that were previously infected with IAV (IAV-mice). Strikingly, naive mice showed no detectable levels of RBD-specific antibodies, demonstrating that preexisting immunity to HA was required in this vaccine-delivery system. Interestingly, the level of systemic  RBD-specific IgG was comparable between IAV-mice immunized with RBD-HA and mice immunized with RBD plus alum (s.c.). Importantly, the level of local  RBD-specific IgA was also comparable between RBD-HA immunized IAV-mice and RBD c-di-GMP–immunized (i.n.) mice. These data collectively indicate that the generation of both systemic and mucosal antibody-specific responses was robust in RBD-HA–immunized IAV-mice. The induction of a potent IgA antibody response in the nasal cavity represents the major advantage of a mucosal vaccine, considering several studies have shown an inverse correlation of mucosal IgA concentrations with the risk of breakthrough SARS-CoV-2 infections –11. ThusThe authors then isolated the antibodies from nasal wash alone and demonstrated that the antibodies were sufficient to neutralize different strains of SARS-CoV-2 in vitro by utilizing pseudotyped viruses displaying Alpha, Delta, and Omicron spike protein. To translate these observations in vivo, they next demonstrated that the nasal RBD-HA immunization in IAV-mice remarkably reduced morbidity and mortality as well as pulmonary viral replication against mouse-adapted SARS-CoV-2 infection. Interestingly, although the authors used an immunization volume that limited the immune response of RBD-HA to the nasal cavity, IAV-mice vaccinated with RBD-HA were also protected against infection in the lower respiratory tract. These data suggest that the protection mediated by RBD-HA in the upper airways prevents viral entry into the lower airways, demonstrating the advantage of mucosal immunization against breakthrough infection rather than only mitigating severe disease. This protection was mediated via the neutralization capacity of RBD-specific antibodies, as the passive transfer of serum from IAV-mice immunized with RBD-HA was capable of mediating protection from lower respiratory tract infection.–/– mice, the authors showed that IAV-IgA was not necessary in the generation of RBD-specific antibodies following RBD-HA administration. Interestingly, preexisting memory CD4+ T cells were also required, as CD4+ T cell–depleting treatment abrogated the generation of RBD-specific antibodies in IAV-mice immunized with RBD-HA. Although these T cells could have been IAV-specific CD4+ memory T cells, prior i.p. administration of IAV-IgG was sufficient to simulate IAV preexisting immunity, which raises a question about the role of bystander CD4+ memory T cells in this process. These findings align with studies reporting an association between effector CD4+ memory T cells and antibody production in COVID-19 convalescent patients . They show that sequential infection with IAV did not alter levels of RBD-specific antibodies following RBD-HA. This result implies that infection history does not affect the efficacy of the RBD-HA vaccine as long as individuals have been exposed to IAV. In the case of countries with low IAV prevalence, the authors show that HA-specific antibodies are sufficient to mimic preexisting immunity. Fourth, the authors demonstrate that preimmunization with the COVID-19 mRNA vaccine can also promote the generation of RBD-specific antibodies. This finding suggests that preexisting immunity to SARS-CoV-2, in the context of immunization and likely infection as well, could be harnessed for the efficient action of an RBD-HA vaccine. Fifth, Kawai et al. show the versatility of their vaccine platform by generating HA fusion vaccines with the N-terminal of SARS-CoV-2 spike protein, the pneumococcal surface protein A of Streptococcus pneumoniae, and the central domain of surface G glycoprotein from RSV. With each fusion vaccine, specific antibodies for the derived antigen were generated and capable of providing protection against subsequent infection. These exciting findings uncover a promising avenue for subunit mucosal vaccines (2 in humans and represent substantial entry points for different pathogens (Kawai and colleagues present the efficacy of their vaccine only in a preclinical study, but this study shows great promise for future translational studies. First, the authors show their immunization model works in both BALB/c and C57BL/6 mice. Second, they provide evidence for the longevity of their immunization approach, showing RBD-specific antibodies were maintained up to six months after immunization. Third, to replicate a diverse history of infections seen in humans, Kawai et al. also utilized two common pulmonary pathogens, vaccines . In factathogens . This vaEmergent strains of SARS-CoV-2 and other pulmonary pathogens are in constant circulation, underscoring the necessity for preventative strategies that curb transmission. The tropism of pulmonary viruses for the upper versus lower respiratory tract dictates the severity of infection. For instance, highly glycosylated seasonal influenza viruses are often eliminated from the airways via the ciliary ladder, which causes a mild disease by limiting the infection to the upper airways. In contrast, the poorly glycosylated 1918 pandemic strain bypasses the upper airways, causing deep lung infection with severe disease . Thus, d


Mild uptake in the left ventricular wall was detected on the oncology 18FDG PET/CT. True myocardiac-involvement could not be distinguished with physiological uptake. The following 68Ga-FAPI-04 showed intense heterogeneous uptake in the left ventricular wall, particularly in the septum and apex area, corresponding with the late gadolinium enhancement regions shown by cardiac MR. Intense uptake was also noted in the mediastinal and bilateral hilar lymph nodes. Endomyocardial biopsy demonstrated sarcoidosis.A 47-year-old female with cardiac dysfunction and lymphadenopathy underwent


DNAM-1 is a major NK cell activating receptor and, together with NKG2D and NCRs, by binding specific ligands, strongly contributes to mediating the killing of tumor or virus-infected cells. DNAM-1 specifically recognizes PVR and Nectin-2 ligands that are expressed on some virus-infected cells and on a broad spectrum of tumor cells of both hematological and solid malignancies. So far, while NK cells engineered for different antigen chimeric receptors (CARs) or chimeric NKG2D receptor have been extensively tested in preclinical and clinical studies, the use of DNAM-1 chimeric receptor-engineered NK cells has been proposed only in our recent proof-of-concept study and deserves further development. The aim of this perspective study is to describe the rationale for using this novel tool as a new anti-cancer immunotherapy. Thert decade .ex vivo amplification methods of NK cells and designs for the expression of various chimeric antigen receptors (CARs) and NKG2D chimeric receptor -human leukocyte antigen (HLA) mismatch , can be receptor , 25 suitIn this context, one should consider that T and NK cells are often dysfunctional in cancer patients, limiting the use of autologous cells for engineered manipulation . NoticeaClinicalTrial.govSo far, the successful use of NK cells engineered for several CARs and for NKG2D chimeric receptor in the hematological and solid tumor settings has been widely reported (in vitro results on the efficacy of never before explored DNAM-1-chimeric receptor-engineered NK cells against neuroblastoma (NB)  . This prvia Src family kinase Fyn and protein kinase C, respectively  is constitutively expressed in T, NK cells, and some myeloid cells. It is a type I transmembrane glycoprotein containing a leader sequence of 18 amino acid (aa), two extracellular Ig-like C2-set domains of 230 aa, a transmembrane domain of 28 aa and a cytoplasmic region of 60 aa. Together with other activating receptors, such as NKG2D and NCRs , DNAM-1 ectively . The cooectively .Adequate expression of DNAM-1 enables NK cells to recognize and kill hematopoietic malignancies such as acute myeloid leukemia (AML) , multiplBoth PVR and Nectin-2 ligands are closely linked to tumorigenesis. Indeed, in addition to being expressed in virus-infected cells , these lThe activating signal mediated by DNAM-1 following the engagement of the ligands PVR or Nectin-2 is counteracted by the competing binding of inhibitory receptors such as TIGIT (T-cell immunoglobulin and ITIM domain) , TACTILEAdoptive transfer of activated NK cells expressing higher and more stable levels of DNAM-1, might be a useful clinical approach to help cancer patients to fight tumor cells. The DNAM-1 chimeric receptor could confer a dual advantage to NK cells: (i) specific recognition of ligands such as PVR and Nectin-2, which are highly expressed in tumor cells, but importantly absent or poorly expressed in normal cells, and (ii) its overexpression, which should result in a favorable molecular imbalance with respect to the normal expression of competing receptors , leading to its increased binding to PVR and Nectin-2. In addition, its function could be strategically improved by in-frame expression of costimulatory molecules that support cytotoxic activity and overcome TME immune escape mechanisms. We previously reported a proof-of-concept study on the activity of DNAM-1-chimeric receptor-engineered NK cells obtained by transient transfection of primary human NK cells for a DNAM-1-chimeric receptor . Specifiin-silico bioinformatic analysis by using GEPIA2  to investigate the prognostic value of PVR and Nectin-2 ligands in a variety of tumor types. We found that higher expression of PVR significantly correlated with lower patient overall survival in ACC, BLCA, COAD, ESCA, HNSC, kidney renal clear cell carcinoma (KIRC), kidney renal papillary cell carcinoma (KIRP), LUAD, LUSC, mesothelioma (MESO), OV, prostate adenocarcinoma (PRAD), SKCM, STAD and uveal melanoma (UVM)  . Similarprotein.atlas.org, GTEx from TCGA database and  production and clinical use of DNAM-1 chimeric receptor-engineered NK cells, primary NK cells should be isolated through leukapheresis by the blood of a HLA-matched unrelated healthy donor, ture use Figure 2ture use , 79, actture use . IdeallyThe adoptive transfer of DNAM-1 chimeric receptor-engineered NK cells is expected to represent an innovative strategic clinical tool to help cancer patients in fighting solid tumors. Therefore, the development of preclinical and clinical studies aimed at obtaining stable, nontoxic, highly antitumor cytotoxic DNAM-1 chimeric receptor-engineered NK cells, in high quantities for cryopreservation and immediate future use, applicable to a broad spectrum of solid tumors, deserves further exploration.The original contributions presented in the study are included in the article/All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.


Magnaporthe oryzae AVRs and ToxB-like). These results demonstrate that the HMA IDs-based design of rice materials with broad and enhanced resistance profiles possesses great application potential but also face considerable challenges.Plant nucleotide-binding and leucine-rich repeat (NLR) proteins are immune sensors that detect pathogen effectors and initiate a strong immune response. In many cases, single NLR proteins are sufficient for both effector recognition and signaling activation. These proteins possess a conserved architecture, including a C-terminal leucine-rich repeat (LRR) domain, a central nucleotide-binding (NB) domain, and a variable N-terminal domain. Nevertheless, many paired NLRs linked in a head-to-head configuration have now been identified. The ones carrying integrated domains (IDs) can recognize pathogen effector proteins by various modes; these are known as sensor NLR (sNLR) proteins. Structural and biochemical studies have provided insights into the molecular basis of heavy metal-associated IDs (HMA IDs) from paired NLRs in rice and revealed the co-evolution between pathogens and hosts by combining naturally occurring favorable interactions across diverse interfaces. Focusing on structural and molecular models, here we highlight advances in structure-guided engineering to expand and enhance the response profile of paired NLR-HMA IDs in rice to variants of the rice blast pathogen MAX-effectors ( However, rice production is constantly threatened by diseases. Magnaporthe oryzae that causes rice blast, also known as “the cancer of rice,” was ranked first in the top 10 list of fungal plant pathogens, annually destroying enough rice to feed more than 200 million people for a year , which recognize avirulence (AVR) effectors and trigger immune responses. In particular, the heavy metal-associated (HMA) IDs in rice have been targeted as candidate-engineering elements for anti-rice blast. Therefore, a deep understanding of the molecular mechanisms of plant–pathogen interactions and further exploration and use of plant immunity for disease control are crucial to promote the sustainable development of green agriculture. In this review, we discuss the immune systems of plants and distinctive structures of NLR pairs. We also highlight the progress made in exploiting the structural basis of interactions between paired NLR-HMA IDs of rice and MAX-effectors (M. oryzae AVRs and ToxB-like) for R gene engineering. Finally, we discuss the prospects that need to be addressed to pave the way for the development of broad-race, spectrum-resistant cultivars.Molecular mechanisms of plant–pathogen interactions have demonstrated that most nucleotide-binding and leucine-rich repeat (NLR) domains are crucial downstream components of host resistance . Plants poptosis . However2via the inhibition of fungal cell wall-degrading enzymes, secretion of fungitoxic peptides and phytoalexins, and cell wall strengthening  and receptor-like proteins (RLPs) function as PRRs that act in the first tier of the plant immune system to respond to PAMPs . The extTo establish successful infections, pathogens have evolved a diverse repertoire of effectors that are delivered to plant cells to interfere with PTI . These eAlthough the two classes of immune receptors involve different activation mechanisms and appear to require different early signaling components, a crosstalk between PTI and ETI appears to be mediated by an integrated signaling network . ETI enh3R genes are inadequate for defense against pathogen invasion and necessitate complementary paired R genes encoding for defense responses  or NAD+ with ATP as substrates to produce ADP-ribosylated adenosine triphosphate (ADPr-ATP) and ADPr-ADPR (di-ADPR) through ADP-ribosylation reactions, which are likely to be further hydrolyzed to pRib-ADP and pRib-AMP (2′-(5′′-phosphoribosyl)-5′-adenosine diphosphate and monophosphate). However, TNLs converge on the enhanced disease susceptibility 1 (EDS1) family of lipase-like proteins, which plays a key link between TNL activation and resistance pathway induction  effector family, which adopts a six β-sandwich fold stabilized by buried hydrophobic residues and commonly, but not always, disulfide bonds  variants . AVR1-COde bonds . Althougde bonds .Ser218,Gly230 binds to AVR-PikDHis46 via hydrogen bonds, forming the core region for the regulation of their interactions. In the Pikm-HMA/AVR-PikD complex, G230 is replaced by Val231 at a structurally equivalent position in the Pikp-HMA structure. Therefore, Pikm-HMAVal231 cannot directly form salt bridges with AVR-PikDHis46. In the Pikm-HMA/AVR-PikE complex, the amino acid at position 46 of the effector protein is asparagine (Asn). Consequently, Asn is rotated out of the binding pocket and is located well away from PikmVal231, and hydrogen bonds are formed between AVR-PikE(Asn46:N<σπ>δ</σπ>2) and both Pikm-HMA(Ser219:OH) and the new water molecule. Such a configuration affects the position of AVR-PikmPhe44-Gly48 and pushes them away from Pikm-HMA IDs, further altering the interactions across the core interface. Such structural changes are related to the decreased binding affinity of AVR-PikE to Pikm-HMA compared with that of AVR-PikD ((Ser219:OH), a considerable difference occurs in orientation. These changes cause the residues of AVR-PikAAsn46 - Pro50 to be located a distance from Pikm-HMA, and such structural observations are again related to decreased effector binding affinity  forms a pre-activation complex with PBS1. The cleavage of PBS1 by AvrPphB  secreted by the pathogenic oomycete Phytophthora infestans leads to conformational changes in PBS1, which serve as a key factor in triggering RPS5-induced HR (RCS2 and PBSTuMV) can also trigger RPS5 activity, thereby expanding the recognition specificity of the host towards pathogens  in the RGA4/RGA5 and Pik-1/Pik2 systems, HMA IDs can act as sNLR, binding MAX-effectors like molecular traps and activating the immune response of the system; (2) RGA5-HMA IDs and Pik-HMA IDs show 54% sequence similarity; (3) structure-function analysis demonstrated high plasticity in HMA ID and MAX-effectors interactions. Because their sequence is unrelated but structurally similar, MAX-effectors bind to different surfaces of structurally conserved HMA IDs. Therefore, the design of HMA IDs with expanded recognition profiles based on HMA IDs/MAX-effector complex structures has immense potential and has achieved promising progress Table 2.Asn261Lys,Lys262Glu, with an expanded AVR-Pik recognition profile for the first time. Besides having enhanced AVR-PikD recognition ability, the mutation could also recognize AVR-PikE and AVR-PikA. Sel1027Val,Gly1009Asp (RGA5-HMA2) mutant with new AVR-Pib recognition sites  (RGA5-HMAm1) and RGA5-HMA (RGA5-HMAm1m2) with expanded MAX-effector recognition profiles (Potato virus X (PVX) expressing GFP or mCherry  genes, enhanced pathogen resistance, and altered plant development, such as dwarfism, are general features of plant autoimmunity (For the structure-guided engineering of immune receptors, sistance . Stackine blight . A five-e blight . Howevere blight . In addiimmunity . Therefoimmunity . Besidesimmunity . NLR misimmunity .6M. oryzae. Therefore, adequate use of the immune systems of crops and in-depth exploration of the interaction modes of NLR proteins and effectors provide an important molecular basis for the mining and designing of R genes and resistant crop varieties. However, the following issues require further consideration and investigation:Coordinated development of structural biology and genetics has provided an important molecular basis for elucidating the interactions between plant NLR proteins and pathogens. In this review article, we summarize the importance of the plant immune system and the progress in genetic engineering strategies for the targeted design of HMA IDs with expanded, enhanced, or new MAX-effector recognition abilities based on the structural information of paired NLR-HMA IDs in rice and MAX-effectors of Transgenic rice varieties conferring increased or extended anti-rice blast specificity by modified HMA IDs are yet to be developed.M. oryzae. It is unclear whether continuous resistance can be provided by transplanting engineered HMA IDs into crops belonging to related or unrelated genera of rice.Exploration of the feasibility of designing HMA IDs through modification or exchange to confer recognition profiles for MAX-effectors of other pathogens besides Elucidation of the molecular mechanisms of synergistic interactions in the paired NLR proteins of rice to determine how paired NLR-HMA IDs activate sNLR proteins to induce molecular transduction of hNLR immune signals.Exploration of the molecular mechanisms and modification strategies for HMA IDs/MAX-effectors as models for further application to other crops for in-depth investigation of the diversity of NLR proteins in recognizing effectors and activating immune mechanisms.XH and JL conceived and designed the review. LG and YM wrote the manuscript. Other authors listed have made a substantial direct and intellectual contribution to the work and approved it for publication.


Multidrug-resistant bacteria pose a significant threat to global public health . In thisThe 11 manuscripts in this research issue revolve around the topic of resistance to carbapenems. Various studies and outbreaks are discussed, highlighting the importance of monitoring and controlling the spread of carbapenem-resistant gram-negative bacteria and other multidrug-resistant organisms in hospital settings. Some studies have developed predictive models and nomograms to identify high-risk patients and personalize risk prediction. Whole-genome sequencing and plasmid characterization techniques have been utilized to analyze the genetic mechanisms of antibiotic resistance and the dissemination of carbapenemases in various bacteria. Increased surveillance and strict infection control measures have been emphasized to contain outbreaks and prevent the further spread of resistance. Overall, the articles demonstrate the urgent need for effective treatment strategies and control measures to combat the growing threat of carbapenem resistance.“Carbapenem-Resistant Gram-Negative Bacterial Infection in Intensive Care Unit Patients: Antibiotic Resistance Analysis and Predictive Model Development” analyzed antibiotic resistance in carbapenem-resistant gram-negative bacteria (CR-GNB) in ICU patients and developed a predictive model . A total of 309 patients with GNB infection were recruited and divided into CR and carbapenem-susceptible (CS) groups. The most prevalent CR-GNB were carbapenem-resistant Klebsiella pneumoniae, carbapenem-resistant Acinetobacter baumannii, and carbapenem-resistant Pseudomonas aeruginosa. A history of combination antibiotic treatments, hospital-acquired infection, and mechanical ventilation ≥ 7 days were independent risk factors for CR-GNB infection. These factors were used to construct a nomogram-based predictive model. The model demonstrated good performance, with an area under the ROC curve (AUC) of 0.753 and 0.718 for the experimental and validation cohort, respectively. The model could be used to guide preventive and treatment measures in identifying patients at high risk of developing CR-GNB infection in the ICU. Another outbreak of multidrug resistant Klebsiella pneumoniae was discussed in the article “Outbreak of colistin resistant, carbapenemase  producing Klebsiella pneumoniae causing blood stream infection among neonates at a tertiary care hospital in India”, which involved 5 out of 7 neonates . The isolates were multidrug resistant, including carbapenems and colistin, and belonged to three different sequence types. The isolates harbored carbapenemase genes and extended-spectrum β-lactamases, but colistin resistance genes could not be detected. K. pneumoniae ST101 was isolated from filtered incubator water and harbored blaNDM-5, blaOXA-232 and ESBL genes, but it was negative for mcr genes. Strict infection control measures were applied, and the outbreak was contained, emphasizing the importance of early detection, surveillance, and proper infection control practices. The study “Detection of IMP-4 and SFO-1 co-producing ST51 Enterobacter hormaechei clinical isolates” investigated the genetic characteristics of Enterobacter hormaechei YQ13422hy and YQ13530hy, two multidrug-resistant clinical isolates co-producing the antibiotic resistance genes blaIMP-4 and blaSFO-1 . Whole-genome sequencing and plasmid characterization techniques were used to identify and analyze the genes and their genetic context. The study highlights the dissemination of blaIMP-4 in E. hormaechei and the transferable IncN-type plasmid carrying the blaIMP-4 resistance gene in this bacterium. It also emphasizes the importance of understanding the genetic mechanisms of antibiotic resistance for developing effective treatment strategies.The study “Development and Validation of Nomograms for Predicting the Risk Probability of Carbapenem Resistance and 28-day All-cause Mortality in Gram-negative Bacteremia among Patients with Hematological Diseases” aimed to develop two nomograms to predict mortality and carbapenem resistance in hospitalized hematological patients with gram-negative bacteria bloodstream infections . The study included 244 patients, and 31.6% of them were resistant to carbapenems. The nomograms were constructed using LASSO regression analysis and multivariate logistic regression analysis, and the models were internally validated. The carbapenem resistance nomogram had a modified C-index of 0.788, while the prognosis nomogram had a modified C-index of 0.873. The decision curve analysis demonstrated that the nomograms were clinically practical for predicting high-risk patients. The study suggests that the nomogram models can be an effective tool for personalized risk prediction in clinical practice. Moreover, a carbapenem-resistant Acinetobacter pittii strain, co-producing chromosomal NDM-1 and OXA-820 carbapenemases, was characterized from a bloodstream infection in the manuscript “Emergence of uncommon KL38-OCL6-ST220 carbapenem resistant Acinetobacter pittii strain, co-producing chromosomal NDM-1 and OXA-820 carbapenemases” . The strain was resistant to imipenem, meropenem, and ciprofloxacin but susceptible to amikacin, colistin, and tigecycline. Whole-genome sequencing revealed the strain contained one circular chromosome and four plasmids, with blaNDM-1 and blaOXA-820 genes located in the chromosome. The strain also contained many virulence factors and 12 prophage regions. Phylogenetic analysis showed that the strain was closely related to an A. pittii strain from Anhui, China. Increased surveillance of this species in hospital and community settings is urgently needed due to the challenge presented by the co-existence of these carbapenemases. In this same context of resistance to carbapenems, the study “Emergence of a Salmonella Rissen ST469 clinical isolate carrying blaNDM-13 in China” describes a variant of the New Delhi metallo-β-lactamase that confers resistance to carbapenems, which was identified in a patient with fever and diarrhea . SR33 was found to be multidrug-resistant and carried many virulence genes. The blaNDM-13 was located in a transmissible IncI1 plasmid and had a conserved genetic context and hybrid promoter. This is the first report of blaNDM-13 in Salmonella, highlighting the need for the monitoring and control of its dissemination. IS1294 may be involved in the movement of blaNDM-13.On the other hand, the study “Genomic transmission analysis of multidrug-resistant Gram-negative bacteria within a newborn unit of a Kenyan Tertiary hospital: a four-month prospective colonization study” conducted in a Kenyan tertiary hospital investigated the prevalence of multidrug-resistant organisms (MDRO) and carbapenem-resistant organisms (CRO) in a newborn unit using routine microbiology, whole-genome sequencing (WGS), and hospital surveillance data . The study included 300 mother–baby pairs and detected MDRO in 16% of neonates at admission, increasing to 44% until discharge, with K. pneumoniae harboring blaNDM-1 and blaNDM-5 being the most frequent CRO. WGS analysis revealed 20 transmission clusters, indicating independent transmission events rather than a large outbreak scenario. The high CRO rate attributed to the spread of NDM-type carbapenemases is a cause for concern.In terms of transmission, the study “Prevalence of carbapenem-resistant hypervirulent Klebsiella pneumoniae and hypervirulent carbapenem-resistant Klebsiella pneumoniae in China determined via mouse lethality tests”  and “Hybrid plasmids encoding antimicrobial resistance and virulence traits among hypervirulent Klebsiella pneumoniae ST2096 in India” . The first study investigated the epidemiology of carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-HvKP) and hypervirulent carbapenem-resistant Klebsiella pneumoniae (Hv-CRKP) in mainland China. The study analyzed 436 Klebsiella pneumoniae strains collected from seven hospitals between 2017 and 2018, using various tests such as sequencing, serotyping, and mouse lethality tests. The study found that the exact prevalence of CR-HvKP is less than 1%, while that of Hv-CRKP is much lower. The authors recommend using mouse lethality tests to accurately determine the prevalence of CR-HvKP and Hv-CRKP.The prevalence and encoding plasmids in hypervirulent resistant organisms was addressed in two studies: The second study genetically characterized a collection of multidrug-resistant hypervirulent Klebsiella pneumoniae (MDR-HvKp) ST2096 isolates carrying both antimicrobial resistance (AMR) and virulence genes on a large hybrid plasmid. The hybrid plasmid carried a CRISPR-cas system, which harbored spacer regions against IncF plasmids, preventing their acquisition. This convergence of virulence and AMR in K. pneumoniae is clinically concerning and highlights the continued emergence of such genotypes globally across the species.“Biofilm formation is not an independent risk factor for mortality in patients with Acinetobacter baumannii bacteremia” . This retrospective study conducted in Taiwan analyzed 711 patients with Acinetobacter baumannii bacteremia, comparing the clinical features of those infected with biofilm-forming and non-biofilm-forming isolates. Multivariate analysis revealed that shock status, higher APACHE II score, lack of appropriate antimicrobial therapy, and carbapenem resistance were independent risk factors for 28-day mortality, but there was no significant difference between the biofilm-forming ability and survival. Patients infected with biofilm-forming isolates had a lower in-hospital mortality rate, and those with congestive heart failure, hematological malignancy, or who received chemotherapy were more likely to be infected with biofilm-forming isolates. Biofilm-forming ability did not influence carbapenem susceptibility.Biofilm formation and mortality has been investigated in “Novel insights related to the rise of KPC-producing Enterobacter cloacae Complex strains within the nosocomial niche” shows that the increase in carbapenem-resistant Enterobacterales has led to the urgent need for new antibiotics . This study analyzed a patient colonized with both K. pneumoniae and ECC strains and in vitro competition assays between high-risk clones of KPC-producing ECC and K. pneumoniae. The results show that these high-risk clones can coexist in the same hospital environment, including the same patient. The findings suggest that the ability of some pandemic clones to compete and occupy a certain niche may explain the worldwide rise of KPC-ECC strains.In Argentina, where Klebsiella pneumoniae and the Enterobacter cloacae complex (ECC) are the two main CRE species, the study All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.