SUMMARY
Carotenoids are important natural pigments that give bright colors to plants. The difference in the accumulation of carotenoids is one of the key factors in the formation of various colors in ...carrot taproots. Carotenoid cleavage dioxygenases (CCDs), including CCD and 9‐cis epoxycarotenoid dioxygenase, are the main enzymes involved in the cleavage of carotenoids in plants. Seven CCD genes have been annotated from the carrot genome. In this study, through expression analysis, we found that the expression level of DcCCD4 was significantly higher in the taproot of white carrot (low carotenoid content) than orange carrot (high carotenoid content). The overexpression of DcCCD4 in orange carrots caused the taproot color to be pale yellow, and the contents of α‐ and β‐carotene decreased sharply. Mutant carrot with loss of DcCCD4 function exhibited yellow color (the taproot of the control carrot was white). The accumulation of β‐carotene was also detected in taproot. Functional analysis of the DcCCD4 enzyme in vitro showed that it was able to cleave α‐ and β‐carotene at the 9, 10 (9′, 10′) double bonds. In addition, the number of colored chromoplasts in the taproot cells of transgenic carrots overexpressing DcCCD4 was significantly reduced compared with that in normal orange carrots. Results showed that DcCCD4 affects the accumulation of carotenoids through cleavage of α‐ and β‐carotene in carrot taproot.
Significance Statement
We analyzed seven carotenoid cleavage dioxygenase genes annotated from the carrot genome. Functional analysis revealed that DcCCD4 catalyzes the degradation of α‐ and β‐carotene to affect carotenoid accumulation. This work may provide a reference for generating new plant varieties with ideal color.
The efficacy of checkpoint immunotherapy to non-small cell lung cancer (NSCLC) largely depends on the tumor microenvironment (TME). Here, we demonstrate that CCL7 facilitates anti-PD-1 therapy for ...the Kras
Tp53
(KP) and the Kras
Lkb1
(KL) NSCLC mouse models by recruiting conventional DC 1 (cDC1) into the TME to promote T cell expansion. CCL7 exhibits high expression in NSCLC tumor tissues and is positively correlated with the infiltration of cDC1 in the TME and the overall survival of NSCLC patients. CCL7 deficiency impairs the infiltration of cDC1 in the TME and the subsequent expansion of CD8
and CD4
T cells in bronchial draining lymph nodes and TME, thereby promoting tumor development in the KP mouse model. Administration of CCL7 into lungs alone or in combination with anti-PD-1 significantly inhibits tumor development and prolongs the survival of KP and KL mice. These findings suggest that CCL7 potentially serves as a biomarker and adjuvant for checkpoint immunotherapy of NSCLC.
The first domesticated carrots were thought to be purple carrots rich in anthocyanins. The anthocyanins biosynthesis in solid purple carrot taproot was regulated by DcMYB7 within P3 region containing ...a gene cluster of six DcMYBs. Here, we described a MYB gene within the same region, DcMYB11c, which was highly expressed in the purple pigmented petioles. Overexpression of DcMYB11c in ‘Kurodagosun’ (KRDG, orange taproot carrot with green petioles) and ‘Qitouhuang’ (QTHG, yellow taproot carrot with green petioles) resulted in deep purple phenotype in the whole carrot plants indicating anthocyanins accumulation. Knockout of DcMYB11c in ‘Deep Purple’ (DPPP, purple taproot carrot with purple petioles) through CRISPR/Cas9‐based genome editing resulted in pale purple phenotype due to the dramatic decrease of anthocyanins content. DcMYB11c could induce the expression of DcbHLH3 and anthocyanins biosynthesis genes to jointly promote anthocyanins biosynthesis. Yeast one‐hybrid assay (Y1H) and dual‐luciferase reporter assay (LUC) revealed that DcMYB11c bound to the promoters of DcUCGXT1 and DcSAT1 and directly activated the expression of DcUCGXT1 and DcSAT1 responsible for anthocyanins glycosylation and acylation, respectively. Three transposons were present in the carrot cultivars with purple petioles but not in the carrot cultivars with green petioles. We revealed the core factor, DcMYB11c, involved in anthocyanins pigmentation in carrot purple petioles. This study provides new insights into precise regulation mechanism underlying anthocyanins biosynthesis in carrot. The orchestrated regulation mechanism in carrot might be conserved across the plant kingdom and useful for other researchers working on anthocyanins accumulation in different tissues.
Summary statement
Here, we described a MYB gene, DcMYB11c, which was highly expressed in the purple pigmented petioles. The orchestrated regulation mechanism in carrot might be conserved across the plant kingdom and useful for other researchers working on anthocyanins accumulation in different tissues.
Network reconnaissance of addresses and ports is prerequisite to a vast majority of cyber attacks. Meanwhile, the static address configuration of networks and hosts simplifies adversarial ...reconnaissance for target discovery. Although the randomization of host addresses has been suggested as a proactive disruption mechanism against such reconnaissance, the proposed approaches do not exploit the full potentials of address randomization in provision of unpredictability and attack adaptability. Moreover, these approaches do not provide thorough analysis on effectiveness and limitations of address randomization against relevant threat models, including stealthy scanning and worms. In this paper, we present an effective address randomization technique, called random host address mutation (RHM), that turns end-hosts into untraceable moving targets. This technique achieves maximum efficacy by allowing address randomization to be highly unpredictable and fast, and adaptive to adversarial behavior, while incurring low operational and reconfiguration overhead. Our approach achieves the following objectives: (1) it achieves high uncertainty in adversary scanning by modeling address mutation randomization as a multi-level satisfiability problem; (2) it adapts the mutation scheme by fast characterization of adversarial reconnaissance patterns; (3) it achieves high mutation rate by separating mutation from end-hosts and managing it via network appliances; and (4) it preserves network integrity, manageability and performance by bounding the size of routing tables, preserving end-to-end reachability, and efficient handling of reconfiguration updates. Our extensive analyses and simulation show that the RHM distorts adversarial reconnaissance, slows down (deters) the attack, and increases its detectability. Consequently, the RHM is effective in countering a significant number of sophisticated threat models, including reconnaissance, stealthy/evasive scanning methods, and targeted attacks. We also address limitations of our approach in terms of effectiveness and applicability.
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Surgery and drug therapy are the two principal options for cancer treatment. However, their clinical benefits are hindered by the difficulty of accurate location of the tumors and ...timely monitoring of the treatment efficacy of drugs, respectively. Rapid development of imaging techniques provides promising tools to address these challenges. Compared with conventional imaging techniques such as magnetic resonance imaging and computed tomography etc., fluorescence imaging exhibits high spatial resolution, real-time imaging capability, and relatively low costs devices. The advancements in fluorescent probes further accelerate the implementation of fluorescence imaging in tumor diagnosis and treatment monitoring. In particular, the emergence of site-specifically activatable fluorescent probes fits the demands of tumor delineation and real-time feedback of the treatment efficacy. A variety of small molecule probes or nanoparticle-based probes have been developed and explored for the above-mentioned applications. This review will discuss recent advances in fluorescent probes with a special focus on activatable nanoprobes and highlight the potential implementation of activatable nanoprobes in fluorescence imaging-guided surgery as well as imaging-guided drug therapy.
The exponential distribution has always been prominent in various disciplines because of its wide range of applications. In this work, a generalization of the classical exponential distribution under ...a neutrosophic environment is scarcely presented. The mathematical properties of the neutrosophic exponential model are described in detail. The estimation of a neutrosophic parameter by the method of maximum likelihood is discussed and illustrated with examples. The suggested neutrosophic exponential distribution (NED) model involves the interval time it takes for certain particular events to occur. Thus, the proposed model may be the most widely used statistical distribution for the reliability problems. For conceptual understanding, a wide range of applications of the NED in reliability engineering is given, which indicates the circumstances under which the distribution is suitable. Furthermore, a simulation study has been conducted to assess the performance of the estimated neutrosophic parameter. Simulated results show that imprecise data with a larger sample size efficiently estimate the unknown neutrosophic parameter. Finally, a complex dataset on remission periods of cancer patients has been analyzed to identify the importance of the proposed model for real-world case studies.
Cancer vaccines have received tremendous attention in cancer immunotherapy due to their capability to induce a tumor-specific immune response. However, their effectiveness is compromised by the ...insufficient spatiotemporal delivery of antigens and adjuvants in the subcellular level to induce a robust CD8+ T cell response. Herein, a cancer nanovaccine G5-pBA/OVA@Mn is prepared through multiple interactions of manganese ions (Mn2+), benzoic acid (BA)-modified fifth generation polyamidoamine (G5-PAMAM) dendrimer, and the model protein antigen ovalbumin (OVA). In the nanovaccine, Mn2+ not only exerts a structural function to assist OVA loading as well as its endosomal escape, but works as an adjuvant of stimulator of interferon genes (STING) pathway. These collaboratively facilitate the orchestrated codelivery of OVA antigen and Mn2+ into cell cytoplasm. Vaccination with G5-pBA/OVA@Mn not only shows a prophylactic effect, but also significantly inhibits growth against B16-OVA tumors, indicating its great potential for cancer immunotherapy.
SUMMARY
The color of purple carrot taproots mainly depends on the anthocyanins sequestered in the vacuoles. Glutathione S‐transferases (GSTs) are key enzymes involved in anthocyanin transport. ...However, the precise mechanism of anthocyanin transport from the cytosolic surface of the endoplasmic reticulum (ER) to the vacuoles in carrots remains unclear. In this study, we conducted a comprehensive analysis of the carrot genome, leading to the identification of a total of 41 DcGST genes. Among these, DcGST1 emerged as a prominent candidate, displaying a strong positive correlation with anthocyanin pigmentation in carrot taproots. It was highly expressed in the purple taproot tissues of purple carrot cultivars, while it was virtually inactive in the non‐purple taproot tissues of purple and non‐purple carrot cultivars. DcGST1, a homolog of Arabidopsis thaliana TRANSPARENT TESTA 19 (TT19), belongs to the GSTF clade and plays a crucial role in anthocyanin transport. Using the CRISPR/Cas9 system, we successfully knocked out DcGST1 in the solid purple carrot cultivar ‘Deep Purple’ (‘DPP’), resulting in carrots with orange taproots. Additionally, DcMYB7, an anthocyanin activator, binds to the DcGST1 promoter, activating its expression. Compared with the expression DcMYB7 alone, co‐expression of DcGST1 and DcMYB7 significantly increased anthocyanin accumulation in carrot calli. However, overexpression of DcGST1 in the two purple carrot cultivars did not change the anthocyanin accumulation pattern or significantly increase the anthocyanin content. These findings improve our understanding of anthocyanin transport mechanisms in plants, providing a molecular foundation for improving and enhancing carrot germplasm.
Significance Statement
These findings improve our understanding of anthocyanin transport mechanisms in plants, providing a molecular foundation for improving and enhancing carrot germplasm.
Background
No comprehensive analysis is available on the viral etiology and clinical characterization among children with severe acute lower respiratory tract infection (SALRTI) in Southern China.
...Methods
Cohort of 659 hospitalized children (2 months to 14 years) with SALRTI admitted to the Pediatric Intensive Care Unit (PICU) in the Guangzhou from May 2015 to April 2018 was enrolled in this study. Nasopharyngeal aspirate specimens or induced sputum were tested for eight categories respiratory viral targets. The viral distribution and its clinical characters were statistically analyzed.
Results
Viral pathogen was detected in 326 (49.5%) of children with SALRTI and there were 36 (5.5%) viral coinfections. Overall, the groups of viruses identified were, in descending order of prevalence: Influenza virus (IFV) (n = 94, 14.3%), respiratory syncytial virus (RSV) (n = 75, 11.4%), human rhinovirus (HRV) (n = 56, 8.5%), adenovirus (ADV) (n = 55, 8.3%), parainfluenza (PIV) (n = 47, 7.1%), human coronavirus (HCoV) (n = 15, 2.3%), human metapneumovirus (HMPV) (n = 14, 2.1%) and human bocavirus (HBoV) (n = 11, 1.7%). The positive rate in younger children (< 5 years) was significantly higher than the positive rate detected in elder children (> 5 years) (52.5% vs 35.1%, P = 0.001). There were clear seasonal peaks for IFV, RSV, HRV, ADV, PIV, and HMPV. And the individuals with different viral infection varied significantly in terms of clinical profiles.
Conclusions
Viral infections are present in a consistent proportion of patients admitted to the PICU. IFV, RSV, HRV, and ADV accounted for more than two‐thirds of all viral SALRTI. Our findings could help the prediction, prevention and potential therapeutic approaches of SALRTI in children.
Highlight
Viral infections are present in a consistent proportion of patients admitted to the Pediatric Intensive Care Unit.
Influenza virus, respiratory syncytial virus, human rhinovirus and adenovirus accounted for more than two‐thirds of all viral SALRTI.
The challenge to improve the clinical efficacy and enlarge the population that benefits from immune checkpoint inhibitors (ICIs) for non‐small‐cell lung cancer (NSCLC) is significant. Based on ...whole‐exosome sequencing analysis of biopsies from NSCLC patients before anti‐programmed cell death protein‐2 (PD‐1) treatment, we identified NLRP4 mutations in the responders with a longer progression‐free survival (PFS). Knockdown of NLRP4 in mouse Lewis lung cancer cell line enhanced interferon (IFN)‐α/β production through the cGAS‐STING‐IRF3/IRF7 axis and promoted the accumulation of intratumoral CD8+ T cells, leading to tumor growth retardation in vivo and a synergistic effect with anti‐PD‐ligand 1 therapy. This was consistent with clinical observations that more tumor‐infiltrating CD8+ T cells and elevated peripheral IFN‐α before receiving nivolumab treatment were associated with a longer PFS in NSCLC patients. Our study highlights the roles of tumor‐intrinsic NLRP4 in remodeling the immune contextures in the tumor microenvironment, making regional type I IFN beneficial for ICI treatment.
Our study addressed novel mechanisms for overcoming failures of immune checkpoint inhibitor (ICI) therapy for non‐small‐cell lung cancer (NSCLC) through endogenous regulation of NLRP4 on type I interferon. This is realized mostly through reshaping the tumor microenvironment to a “hot” tumor status, which is beneficial to the treatment of anti‐programmed cell death protein‐1. Furthermore, we provided new evidence on type I interferon (regulated by NLRP4 in our study) as a potential immune adjuvant to improve the efficacy of NSCLC patients ineffective to ICI treatment.
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