Knockdown of NR4A1 alleviates doxorubicin-induced cardiotoxicity through inhibiting the activation of the NLRP3 inflammasome.

Doxorubicin (DOX) is a widely used antitumor drug, but its clinical applicability is hampered by the unfortunate side effect of DOX-induced cardiotoxicity (DIC). In our current study, we retrieved three high-throughput sequencing datasets related to DIC from the Gene Expression Omnibus (GEO) datasets. We conducted differential analysis using R (DESeq2) to pinpoint differentially expressed genes (DEGs, and identified 11 genes that were consistently altered in both the control and DOX-treated groups. Notably, our Random Forest analysis of these three GEO datasets highlighted the significance of nuclear receptor subfamily 4 group A member 1 (NR4A1) in the context of DIC. The DOX-induced mouse model and cell model were used for the in vivo and in vitro studies to reveal the role of NR4A1 in DIC. We found that silencing NR4A1 by adeno-associated virus serotype 9 (AAV9) contained shRNA in vivo alleviated the DOX-induced cardiac dysfunction, cardiomyocyte injury and fibrosis. Mechanistically, we found NR4A1 silencing was able to inhibit DOX-induced the cleavage of NLRP3, IL-1ß and GSDMD in vivo. Further in vitro studies have shown that inhibition of NR4A1 suppressed DOX-induced cytotoxicity and oxidative stress through the same molecular mechanism. We prove that NR4A1 plays a critical role in DOX-induced cardiotoxicity by inducing pyroptosis via activation of the NLRP3 inflammasome, and it might be a promising therapeutic target for DIC.

Adhesion between EVs and tumor cells facilitated EV-encapsulated doxorubicin delivery via ICAM1.

Doxorubicin (Dox) is an anti-tumor drug with a broad spectrum, whereas the cardiotoxicity limits its further application. In clinical settings, liposome delivery vehicles are used to reduce Dox cardiotoxicity. Here, we substitute extracellular vesicles (EVs) for liposomes and deeply investigate the mechanism for EV-encapsulated Dox delivery. The results demonstrate that EVs dramatically increase import efficiency and anti-tumor effects of Dox in vitro and in vivo, and the efficiency increase benefits from its unique entry pattern. Dox-loading EVs repeat a "kiss-and-run" motion before EVs internalization. Once EVs touch the cell membrane, Dox disassociates from EVs and directly enters the cytoplasm, leading to higher and faster Dox import than single Dox. This unique entry pattern makes the adhesion between EVs and cell membrane rather than the total amount of EV internalization the key factor for regulating the Dox import. Furthermore, we recognize ICAM1 as the molecule mediating the adhesion between EVs and cell membranes. Interestingly, EV-encapsulated Dox can induce ICAM1 expression by irritating IFN-γ and TNF-α secretion in TME, thereby increasing tumor targeting of Dox-loading EVs. Altogether, EVs and EV-encapsulated Dox synergize via ICAM1, which collectively enhances the curative effects for tumor treatment.

NLRP3 expression and its predictive role in preserved ejection fraction among non-valvular atrial fibrillation patients.

INTRODUCTION: To investigate the expression and predictive value of NOD-like receptor thermoprotein domain-related protein 3 (NLRP3) in patients with nonvalvular atrial fibrillation (NVAF) with ejection fraction preserved heart failure (HFpEF). METHODS: A retrospective analysis of 121 patients diagnosed with NVAF. According to the occurrence of HFpEF, 81 patients was assigned in the NVAF group and 40 patients in the NVAF/HFpEF group. The levels of NLRP3, B natriuretic peptide (BNP), and interleukin-1ß were determined using enzyme-linked immunosorbent assay (ELISA). Independent predictors for HFpEF in NVAF was determined using logistic regression. The receiver operating characteristic curve (ROC) was used to evaluate the predictive value of each factor. RESULTS: Expression levels of NLRP3, BNP, and IL-1ß in the NVAF/HFpEF group, as well as the H2FPEF score was significantly higher than those in the NVAF group. Pearson analysis showed that NLRP3, BNP, and IL-1ß expression levels in NVAF patients and H2FPEF score was positively correlated (r=0.409, r=0.244, r=0.299, p < 0.001). Multivariate logistic regression analysis showed that NLRP3, BNP, or H2FPEF score can be used as independent factor for predicting the occurrence of HFpEF in NVAF. ROC curves showed that the area under the curve (AUC) of NLRP3, BNP, and H2FPEF score for predicting the occurrence of HFpEF in NVAF patients were 0.856, 0.831 and 0.811, respectively. CONCLUSION: NLRP3 level is elevated in the peripheral blood of NVAF patients with HFpEF and is positively correlated with the H2FPEF score. NLRP3 may serve as a potential predictor of HFpEF in patients with NVAF.

Effect of histidine covalent modification on strigolactone receptor activation and selectivity.

The parasitic weed Striga has led to billions of dollars' worth of agricultural productivity loss worldwide. Striga detects host plants using compounds of the strigolactone class of phytohormones. Early steps in the strigolactone signaling pathway involve substrate binding and hydrolysis followed by a conformational change to an "active" or "closed" state, after which it associates with a MAX2-family downstream signaling partner. The structures of the inactive and active states of strigolactone receptors are known through X-ray crystallography, and the transition pathway from the inactive to active state in apo receptors has previously been characterized using molecular dynamics simulations. However, it also has been suggested that a covalent butenolide modification of the receptor on the catalytic histidine through substrate hydrolysis promotes formation of the active state. Using molecular dynamics simulations, we show that the presence of the covalent butenolide enhances activation in both AtD14, a receptor found in Arabidopsis, and ShHTL7, a receptor found in Striga, but the enhancement is ∼50 times greater in ShHTL7. We also show that several conserved interactions with the covalent butenolide modification promote transition to the active state in both AtD14 (non-parasite) and ShHTL7 (parasite). Finally, we demonstrate that the enhanced activation of ShHTL7 likely results from disruption of ShHTL7-specific histidine interactions that inhibited activation in the apo case. These results provide a possible explanation for difference in strigolactone sensitivity seen between different strigolactone-sensitive proteins and can be used to aid the design of selective modulators to control Striga parasites.

GPX3 is a key cholesterol-related gene associated with prognosis and tumor-infiltrating T cells in colorectal cancer.

High cholesterol is an important factor inducing colorectal cancer (CRC). The study aims to determine the key genes and regulatory mechanism associated with tumor-infiltrating T cells underlying cholesterol-induced CRC. Gene expression data and clinical data from CRCS in The Cancer Genome Atlas (TCGA) were selected for differential expression and survival analysis. A total of 5,815 DEGs and 21 cholesterol-associated KEGG pathways were identified. Subsequently, 128 CRCs and 127 patients without obvious intestinal lesions were recruited to analyze the relationship between GPX3 expression, cholesterol levels, and pathologic condition. The results showed that the expression of cholesterol-related gene GPX3 was negatively associated with cholesterol level, but positively correlated with Ki-67 proliferation index in CRC. The expression of GPX3 was higher in CRC patients who were in poorly differentiated and advanced stage. In addition, a mice model of high-cholesterol diet intervention was constructed to detect the levels of cholesterol and GPX3 in the peripheral blood of mice, and it was found that the expression level of GPX3 in high-cholesterol mice was lower than that in normal diet mice. CD8+ T cells were isolated from the spleen of mice and the T cell surface receptors were detected. It was found that the expression of CD69 in CD8+ T cells of mice interfered with the high-cholesterol diet, while the expression of PD1, TIM-3, and CTLA-4 was increased. CD8+ T cells were co-cultured with MC38 cells to detect the proliferation rate of CRC cells. The results showed that the tumor cell proliferation ratio in the high cholesterol group was higher than that in the control group. Furthermore, GPX3 downstream genes associated with m6A modification and tumor-infiltrating T cells were screened, and a T cell immune-related ceRNA network was constructed. In total, 53 GPX3 downstream genes associated with m6A modification and tumor-infiltrating T cells were identified. A PPI network that contained 45 nodes and 85 interaction pairs was constructed. The ceRNA network, including 39 miRNA-target and 43 lncRNA-miRNA regulatory pairs, was constructed. In conclusion, GPX3 is a potential target for cholesterol regulation of T cell immunity in CRC.

Integration of machine learning with computational structural biology of plants.

Computational structural biology of proteins has developed rapidly in recent decades with the development of new computational tools and the advancement of computing hardware. However, while these techniques have widely been used to make advancements in human medicine, these methods have seen less utilization in the plant sciences. In the last several years, machine learning methods have gained popularity in computational structural biology. These methods have enabled the development of new tools which are able to address the major challenges that have hampered the wide adoption of the computational structural biology of plants. This perspective examines the remaining challenges in computational structural biology and how the development of machine learning techniques enables more in-depth computational structural biology of plants.

Role of substrate recognition in modulating strigolactone receptor selectivity in witchweed.

Witchweed, or Striga hermonthica, is a parasitic weed that destroys billions of dollars' worth of crops globally every year. Its germination is stimulated by strigolactones exuded by its host plants. Despite high sequence, structure, and ligand-binding site conservation across different plant species, one strigolactone receptor in witchweed, ShHTL7, uniquely exhibits a picomolar EC50 for downstream signaling. Previous biochemical and structural analyses have hypothesized that this unique ligand sensitivity can be attributed to a large binding pocket volume in ShHTL7 resulting in enhanced ability to bind substrates, but additional structural details of the substrate-binding process would help explain its role in modulating the ligand selectivity. Using long-timescale molecular dynamics simulations, we demonstrate that mutations at the entrance of the binding pocket facilitate a more direct ligand-binding pathway to ShHTL7, whereas hydrophobicity at the binding pocket entrance results in a stable "anchored" state. We also demonstrate that several residues on the D-loop of AtD14 stabilize catalytically inactive conformations. Finally, we show that strigolactone selectivity is not modulated by binding pocket volume. Our results indicate that while ligand binding is not the sole modulator of strigolactone receptor selectivity, it is a significant contributing factor. These results can be used to inform the design of selective antagonists for strigolactone receptors in witchweed.

Correction: Genetics, Receptor Binding Property, and Transmissibility in Mammals of Naturally Isolated H9N2 Avian Influenza Viruses.

[This corrects the article DOI: 10.1371/journal.ppat.1004508.].

Novel statistics predict the COVID-19 pandemic could terminate in 2022.

Many people want to know when the COVID-19 pandemic will end and life will return to normal. This question is highly elusive and distinct predictions have been proposed. In this study, the global mortality and case fatality rate of COVID-19 were analyzed using nonlinear regression. The analysis showed that the COVID-19 pandemic could terminate in 2022, but COVID-19 could be one or two times more deadly than seasonal influenza by 2023. The prediction considered the possibility of the emergence of new variants of SARS-CoV-2 and was supported by the features of the Omicron variant and other facts. As the herd immunity against COVID-19 established through natural infections and mass vaccination is distinct among countries, COVID-19 could be more or less deadly in some countries in the coming years than the prediction. Although the future of COVID-19 will have multiple possibilities, this statistics-based prediction could aid to make proper decisions and establish an example on the prediction of infectious diseases.

Should the world collaborate imminently to develop neglected live-attenuated vaccines for COVID-19?

The rapid spread of the Delta variant suggests that SARS-CoV-2 will likely be rampant for months or years and could claim millions of more lives. All the known vaccines cannot well defeat SARS-CoV-2 due to their limited efficacy and production efficiency, except for the neglected live-attenuated vaccines (LAVs), which could have a much higher efficacy and much higher production efficiency than other vaccines. LAVs, like messiahs, have defeated far more pathogenic viruses than other vaccines in history, and most current human vaccines for viral diseases are safe LAVs. LAVs can block completely infection and transmission of relevant viruses and their variants. They can hence inhibit the emergence of vaccine-escape and virulence-enhancing variants and protect immunologically abnormal individuals better in general. The safety of COVID-19 LAVs, which could save millions of more lives, can be solidly guaranteed through animal experiments and clinical trials. The safety of COVID-19 LAVs could be greatly enhanced with intramuscular or oral administration, or administration along with humanized neutralizing monoclonal antibodies. Together, extensive global collaboration, which can greatly accelerate the development of safe COVID-19 LAVs, is imminently needed.

A highly powerful nonspecific strategy to reduce COVID-19 deaths.

The coronavirus disease 2019 (COVID-19) pandemic caused by the coronavirus severe acute respiratory syndrome coronavirus 2 remains risky worldwide. We elucidate here that good IDM (isolation, disinfection, and maintenance of health) is powerful to reduce COVID-19 deaths based on the striking differences in COVID-19 case fatality rates among various scenarios. IDM means keeping COVID-19 cases away from each other and from other people, disinfecting their living environments, and maintaining their health through good nutrition, rest, and treatment of symptoms and pre-existing diseases (not through specific antiviral therapy). Good IDM could reduce COVID-19 deaths by more than 85% in 2020 and more than 99% in 2022. This is consistent with the fact that good IDM can minimize co-infections and maintain body functions and the fact that COVID-19 has become less pathogenic (this fact was supported with three novel data in this report). Although IDM has been frequently implemented worldwide to some degree, IDM has not been highlighted sufficiently. Good IDM is relative, nonspecific, flexible, and feasible in many countries, and can reduce deaths of some other relatively mild infectious diseases. IDM, vaccines, and antivirals aid each other to reduce COVID-19 deaths. The IDM concept and strategy can aid people to improve their health behavior and fight against COVID-19 and future pandemics worldwide.

Highly adaptive Phenuiviridae with biomedical importance in multiple fields.

The newly established virus family Phenuiviridae in Bunyavirales harbors viruses infecting three kingdoms of host organisms (animals, plants, and fungi), which is rare in known virus families. Many phenuiviruses are arboviruses and replicate in two distinct hosts (e.g., insects and humans or rice). Multiple phenuivirid species, such as Dabie bandavirus, Rift Valley fever phlebovirus, and Rice stripe tenuivirus, are highly pathogenic to humans, animals, or plants. They impose heavy global burdens on human health, livestock industry, and agriculture and are research hotspots. In recent years the taxonomy of Phenuiviridae has been expanded greatly, and research on phenuiviruses has made significant progress. With these advances, this review drew a novel panorama regarding the biomedical significance, distribution, morphology, genomics, taxonomy, evolution, replication, transmission, pathogenesis, and control of phenuiviruses, to aid researchers in various fields to recognize this highly adaptive and important virus family and conduct relevant risk analysis.

All-optically modulated nonvolatile optical switching based on a graded-index multimode fiber.

Photonic switches have attractive application prospects in optical communication data networks that require dynamic reconfiguration. Integrating optical switching devices with optical fiber, the most widely deployed photonic technology platform, can realize signal transmission and processing in practical applications. Here, we demonstrate the multilevel optical switching using the phase-change material Ge2Sb2Te5 (GST) integrated on a graded-index multimode fiber. This switching process works by exploiting the significant difference in extinction coefficient between the crystalline state and the amorphous state of the GST. Using GST to achieve the switch function, no external energy source is needed to maintain the existing state of the switch, and the device is nonvolatile. This multi-level optical switch is an all-fiber integrated device. We apply GST to the end facets of the graded-index multimode fiber by magnetron sputtering, which is a reflective structure. A pulsing scheme is used to control the optical propagation state of the optical modulation signal to realize the switching function. It can store up to 11 non-volatile reliable and repeatable levels encoded by the pump source laser with a wavelength of 1550 nm. At the same time, the switching process between states is on the order of hundreds of nanoseconds. The present experimental results demonstrate the feasibility of 11 multilevel states in the field of optical fibers commonly used in communications. It can be well coupled with the all-fiber terminal device. It also shows that the device is still applicable in the 1525 nm∼1610 nm broadband range, promising for designing future multilevel photonic switches and memory devices.

Off-label underdosing of four individual NOACs in patients with nonvalvular atrial fibrillation: A systematic review and meta-analysis of observational studies.

BACKGROUND: Although several meta-analyses have examined the effects of off-label underdosing of nonvitamin K antagonist oral anticoagulants (NOACs) compared with their recommended doses in patients with atrial fibrillation (AF), they combined different kinds of NOACs in their primary analyses. Herein, we first conducted a meta-analysis to separately assess the effects of off-label underdosing versus on-label dosing of four individual NOACs on adverse outcomes in the AF population. METHODS: The PubMed and Embase database were systemically searched until November 2021 to identify the relevant studies. Adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) were pooled by utilizing a random-effects model. RESULTS: A total of nine studies with 144,797 patients taking NOACs were included in the meta-analysis. In the pooled analysis, off-label underdosing of rivaroxaban was related to an increased risk of stroke or systemic embolism (HR = 1.31, 95% CI 1.05-1.63; p = .02), whereas off-label underdosing of apixaban was associated with a higher risk of all-cause death (HR = 1.21, 95% CI 1.05-1.40; p = .01). When comparing off-label underdosing versus on-label dosing of dabigatran or edoxaban, no differences were found in the primary and secondary clinical outcomes. CONCLUSION: Off-label underdosing of rivaroxaban may increase the risk of stroke or systematic embolism, whereas off-label underdosing of apixaban may heighten the incidence of all-cause death.

All-fiber nonvolatile broadband optical switch using an all-optical method.

Optical switches based on phase change materials have enormous application potential in optical logic circuits and optical communication systems. Integration of all-optical switching devices with optical fibers is a promising approach for realizing practical applications in enabling the optical fiber to transmit and process signals simultaneously. We describe an all-fiber nonvolatile broadband optical switch using an all-optical method. We use a single optical pulse to modulate the phase change material deposited on the tapered fiber to achieve logical control of the transmitted light. The response time of our optical switch is 80 ns for SET and 200 ns for RESET. Our optical switches can operate in the C-band (1530-1565 nm). The optical switching contrast is 40%. Our approach paves the way for all-optical nonvolatile fiber optic communication.

Activation Mechanism of Strigolactone Receptors and Its Impact on Ligand Selectivity between Host and Parasitic Plants.

Parasitic weeds such as Striga have led to significant losses in agricultural productivity worldwide. These weeds use the plant hormone strigolactone as a germination stimulant. Strigolactone signaling involves substrate hydrolysis followed by a conformational change of the receptor to a "closed" or "active" state that associates with a signaling partner, MAX2/D3. Crystal structures of active and inactive AtD14 receptors have helped elucidate the structural changes involved in activation. However, the mechanism by which the receptor activates remains unknown. The ligand dependence of AtD14 activation has been disputed by mutagenesis studies showing that enzymatically inactive receptors are able to associate with MAX2 proteins. Furthermore, activation differences between strigolactone receptor in Striga, ShHTL7, and AtD14 could contribute to the high sensitivity to strigolactones exhibited by parasitic plants. Using molecular dynamics simulations, we demonstrate that both AtD14 and ShHTL7 could adopt an active conformation in the absence of ligand. However, ShHTL7 exhibits a higher population in the inactive apo state as compared to the AtD14 receptor. We demonstrate that this difference in inactive state population is caused by sequence differences between their D-loops and interactions with the catalytic histidine that prevent full binding pocket closure in ShHTL7. These results indicate that ligand hydrolysis would enhance the active state population by destabilizing the inactive state in ShHTL7 as compared to AtD14. We also show that the mechanism of activation is more concerted in AtD14 than in ShHTL7 and that the main barrier to activation in ShHTL7 is closing of the binding pocket.

Efficacy and safety of electrical stimulation for stress urinary incontinence in women: a systematic review and meta-analysis.

INTRODUCTION AND HYPOTHESIS: This systematic review and meta-analysis was aimed at investigating the safety and short- and long-term efficacy of electrical stimulation (ES) in women with stress urinary incontinence (SUI). METHODS: PubMed, Embase, and the Cochrane database were searched for randomized controlled trials (RCTs) conducted up to 2020. Studies comparing ES with sham ES or no intervention were included. Standardized mean differences (SMDs), weighted mean differences (WMDs), relative risks (RR), and 95% confidence intervals (CIs) were calculated. RESULTS: This study included 9 RCTs, involving a total of 982 patients, of whom 520 received ES. Our results showed that in the short term (< 3 months), compared with sham ES or no intervention, ES significantly improved incontinence-specific quality of life (IQOL) (p = 0.003; SMD = 0.90 [95% CI, 0.30 to 1.50]; I2 = 88%) and reduced urine leakage (p < 0.00001; WMD = -6.15 [95% CI, -8.29 to -4.01]; I2 = 0%) but did not significantly reduce the frequency of incontinence episodes (p = 0.34; WMD = -0.98 [95% CI, -2.99 to 1.04]; I2 = 85%). In the long term (3-7.5 months), ES significantly improved IQOL (p = 0.0009; SMD = 1.14 [95% CI, 0.47 to 1.81]; I2 = 91%) and reduced the frequency of incontinence episodes (p = 0.0009; WMD = -2.45 [95% CI, -3.90 to -1.01]; I2 = 79%) but did not significantly reduce urine leakage (p = 0.27; WMD = -9.21 [95% CI, -25.57 to 7.14]; I2 = 71%). There was no significant difference in adverse events between ES and sham ES or no intervention (p = 0.36; RR = 1.34 [95% CI, 0.72 to 2.50]; I2 = 0%). A test for subgroup differences showed that electroacupuncture (EA) improved long-term IQOL to a greater extent than vaginal ES (VES), whereas there was no significant difference in efficacy between EA and VES in short- or long-term reduction of urine leakage, frequency of incontinence episodes, or short-term IQOL improvement. CONCLUSIONS: ES may improve short- and long-term IQOL for women with SUI, but it appears to provide only short-term reduction in urine leakage and long-term reduction in frequency of incontinence episodes. However, we cannot draw any conclusion on the safety between ES and sham ES or no intervention because of the rarity of adverse events. It is still uncertain whether EA is comparable or superior to VES owing to an insufficient number of studies and patients. The conclusions should be considered carefully because of the limited quality and quantity of the RCTs included. Further rigorous RCTs with adequate sample sizes and long follow-up are necessary to fully validate our findings.

A tensor-based framework for studying eigenvector multicentrality in multilayer networks.

Centrality is widely recognized as one of the most critical measures to provide insight into the structure and function of complex networks. While various centrality measures have been proposed for single-layer networks, a general framework for studying centrality in multilayer networks (i.e., multicentrality) is still lacking. In this study, a tensor-based framework is introduced to study eigenvector multicentrality, which enables the quantification of the impact of interlayer influence on multicentrality, providing a systematic way to describe how multicentrality propagates across different layers. This framework can leverage prior knowledge about the interplay among layers to better characterize multicentrality for varying scenarios. Two interesting cases are presented to illustrate how to model multilayer influence by choosing appropriate functions of interlayer influence and design algorithms to calculate eigenvector multicentrality. This framework is applied to analyze several empirical multilayer networks, and the results corroborate that it can quantify the influence among layers and multicentrality of nodes effectively.

SARS-CoV-2 replicating in nonprimate mammalian cells probably have critical advantages for COVID-19 vaccines due to anti-Gal antibodies: A minireview and proposals.

Glycoproteins of enveloped viruses replicating in nonprimate mammalian cells carry α-1,3-galactose (α-Gal) glycans, and can bind to anti-Gal antibodies which are abundant in humans. The antibodies have protected humans and their ancestors for millions of years, because they inhibit replication of many kinds of microbes carrying αGal glycans and aid complements and macrophages to destroy them. Therefore, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replicating in nonprimate mammalian cells (eg, PK-15 cells) carry αGal glycans and could be employed as a live vaccine for corona virus 2019 (COVID-19). The live vaccine safety could be further enhanced through intramuscular inoculation to bypass the fragile lungs, like the live unattenuated adenovirus vaccine safely used in US recruits for decades. Moreover, the immune complexes of SARS-CoV-2 and anti-Gal antibodies could enhance the efficacy of COVID-19 vaccines, live or inactivated, carrying α-Gal glycans. Experiments are imperatively desired to examine these novel vaccine strategies which probably have the critical advantages for defeating the pandemic of COVID-19 and preventing other viral infectious diseases.

Live unattenuated vaccines for controlling viral diseases, including COVID-19.

Live unattenuated vaccines (LUVs) have been neglected for decades, due to widespread prejudice against their safety, even though they have successfully controlled yellow fever and adenovirus infection in humans as well as rinderpest and infectious bursal disease in animals. This review elucidated that LUVs could be highly safe with selective use of neutralizing antivirus antibodies, natural antiglycan antibodies, nonantibody antivirals, and ectopic inoculation. Also, LUVs could be of high efficacy, high development speed, and high production efficiency, with the development of humanized monoclonal antibodies and other modern technologies. They could circumvent antibody-dependent enhancement and maternal-derived antibody interference. With these important advantages, LUVs could be more powerful than other vaccines for controlling some viral diseases, and they warrant urgent investigation with animal experiments and clinical trials for defeating the COVID-19 pandemic caused by the novel coronavirus SARS-CoV-2.