The subcommissural organ regulates brain development via secreted peptides.

The subcommissural organ (SCO) is a gland located at the entrance of the aqueduct of Sylvius in the brain. It exists in species as distantly related as amphioxus and humans, but its function is largely unknown. Here, to explore its function, we compared transcriptomes of SCO and non-SCO brain regions and found three genes, Sspo, Car3 and Spdef, that are highly expressed in the SCO. Mouse strains expressing Cre recombinase from endogenous promoter/enhancer elements of these genes were used to genetically ablate SCO cells during embryonic development, resulting in severe hydrocephalus and defects in neuronal migration and development of neuronal axons and dendrites. Unbiased peptidomic analysis revealed enrichment of three SCO-derived peptides, namely, thymosin beta 4, thymosin beta 10 and NP24, and their reintroduction into SCO-ablated brain ventricles substantially rescued developmental defects. Together, these data identify a critical role for the SCO in brain development.

The subcommissural organ regulates brain development via secreted peptides.

The subcommissural organ (SCO) is a gland located at the entrance of the aqueduct of Sylvius in the brain. It exists in species as distantly related as amphioxus and humans, but its function is largely unknown. To explore its function, we compared transcriptomes of SCO and non-SCO brain regions and found three genes, Sspo, Car3, and Spdef, that are highly expressed in the SCO. Mouse strains expressing Cre recombinase from endogenous promoter/enhancer elements of these genes were used to genetically ablate SCO cells during embryonic development, resulting in severe hydrocephalus and defects in neuronal migration and development of neuronal axons and dendrites. Unbiased peptidomic analysis revealed enrichment of three SCO-derived peptides, namely thymosin beta 4, thymosin beta 10, and NP24, and their reintroduction into SCO-ablated brain ventricles substantially rescued developmental defects. Together, these data identify a critical role for the SCO in brain development.

Ultrasound-augmented cancer immunotherapy.

Cancer is a growing worldwide health problem with the most broadly studied treatments, in which immunotherapy has made notable advancements in recent years. However, innumerable patients have presented a poor response to immunotherapy and simultaneously experienced immune-related adverse events, with failed therapeutic results and increased mortality rates. Consequently, it is crucial to develop alternate tactics to boost therapeutic effects without producing negative side effects. Ultrasound is considered to possess significant therapeutic potential in the antitumor field because of its inherent characteristics, including cavitation, pyrolysis, and sonoporation. Herein, this timely review presents the comprehensive and systematic research progress of ultrasound-enhanced cancer immunotherapy, focusing on the various ultrasound-related mechanisms and strategies. Moreover, this review summarizes the design and application of current sonosensitizers based on sonodynamic therapy, with an attempt to provide guidance on new directions for future cancer therapy.

The relationship between sarcopenia and metabolic dysfunction-associated fatty liver disease among the young and middle-aged populations.

BACKGROUND: Metabolic dysfunction-associated fatty liver disease (MAFLD) has been proposed as a new term for diagnosing fatty liver disease, which is considered to be a multi-systemic disease with multiple extrahepatic manifestations, including sarcopenia. The link between sarcopenia and MAFLD remains uncertain, especially among young and middle-aged adults. Thus, we examined the relationship between MAFLD and sarcopenia in young and middle-aged individuals in this study. METHODS: A total of 2214 individuals with laboratory tests, dual-energy X-ray absorptiometry and ultrasound transient elastography from NHANES 2017-2018 were selected for this study. MAFLD was diagnosed as fatty liver disease with any one of the situations: overweight/obesity, diabetes mellitus, presence of metabolic dysregulation. Sarcopenia was defined by appendicular lean mass adjusted for body mass index (BMI). Multivariable logistic regression and restricted cubic spline (RCS) model were applied to explore the relationship between MAFLD and sarcopenia, and the mediation analyses were also conducted. Moreover, subgroup analyses stratified by BMI and lifestyles were done. RESULTS: The prevalence of MAFLD was 47.85%, and nearly 8.05% of participants had sarcopenia. The prevalence of sarcopenia was higher in participants with MAFLD (12.75%; 95% CI 10.18-15.31%) than in the non-MAFLD (3.73%; 95% CI 2.16-5.31%). MAFLD was significantly positively associated with sarcopenia after adjustments [OR = 2.87 (95% CI: 1.62-5.09)]. Moreover, significant positive associations were observed between liver fibrosis and sarcopenia prevalence in MAFLD patients (OR = 2.16; 95% CI 1.13-4.15). The RCS curve revealed that MAFLD was linearly associated with sarcopenia. The relationship between the MAFLD and sarcopenia were mediated by C-reactive protein (mediation proportion: 15.9%) and high-density lipoprotein cholesterol (mediation proportion: 18.9%). Subgroup analyses confirmed the association between MAFLD and sarcopenia differed in different lifestyle groups. CONCLUSIONS: Both MAFLD prevalence and severity was significantly associated with sarcopenia. Thus, clinicians should advise comorbidity screening and lifestyle changes to young and middle-aged patients.

Structure of the human ATP synthase.

Biological energy currency ATP is produced by F1Fo-ATP synthase. However, the molecular mechanism for human ATP synthase action remains unknown. Here, we present snapshot images for three main rotational states and one substate of human ATP synthase using cryoelectron microscopy. These structures reveal that the release of ADP occurs when the ß subunit of F1Fo-ATP synthase is in the open conformation, showing how ADP binding is coordinated during synthesis. The accommodation of the symmetry mismatch between F1 and Fo motors is resolved by the torsional flexing of the entire complex, especially the γ subunit, and the rotational substep of the c subunit. Water molecules are identified in the inlet and outlet half-channels, suggesting that the proton transfer in these two half-channels proceed via a Grotthus mechanism. Clinically relevant mutations are mapped to the structure, showing that they are mainly located at the subunit-subunit interfaces, thus causing instability of the complex.

Structure of the human respiratory complex II.

Human complex II is a key protein complex that links two essential energy-producing processes: the tricarboxylic acid cycle and oxidative phosphorylation. Deficiencies due to mutagenesis have been shown to cause mitochondrial disease and some types of cancers. However, the structure of this complex is yet to be resolved, hindering a comprehensive understanding of the functional aspects of this molecular machine. Here, we have determined the structure of human complex II in the presence of ubiquinone at 2.86 Å resolution by cryoelectron microscopy, showing it comprises two water-soluble subunits, SDHA and SDHB, and two membrane-spanning subunits, SDHC and SDHD. This structure allows us to propose a route for electron transfer. In addition, clinically relevant mutations are mapped onto the structure. This mapping provides a molecular understanding to explain why these variants have the potential to produce disease.

Somatic variants of MAP3K3 are sufficient to cause cerebral and spinal cord cavernous malformations.

Cerebral cavernous malformations (CCMs) and spinal cord cavernous malformations (SCCMs) are common vascular abnormalities of the CNS that can lead to seizure, haemorrhage and other neurological deficits. Approximately 85% of patients present with sporadic (versus congenital) CCMs. Somatic mutations in MAP3K3 and PIK3CA were recently reported in patients with sporadic CCM, yet it remains unknown whether MAP3K3 mutation is sufficient to induce CCMs. Here we analysed whole-exome sequencing data for patients with CCM and found that ∼40% of them have a single, specific MAP3K3 mutation [c.1323C>G (p.Ile441Met)] but not any other known mutations in CCM-related genes. We developed a mouse model of CCM with MAP3K3I441M uniquely expressed in the endothelium of the CNS. We detected pathological phenotypes similar to those found in patients with MAP3K3I441M. The combination of in vivo imaging and genetic labelling revealed that CCMs were initiated with endothelial expansion followed by disruption of the blood-brain barrier. Experiments with our MAP3K3I441M mouse model demonstrated that CCM can be alleviated by treatment with rapamycin, the mTOR inhibitor. CCM pathogenesis has usually been attributed to acquisition of two or three distinct genetic mutations involving the genes CCM1/2/3 and/or PIK3CA. However, our results demonstrate that a single genetic hit is sufficient to cause CCMs.

The direct catalytic synthesis of ultrasmall Cu2O-coordinated carbon nitrides on ceria for multimodal antitumor therapy.

Engineering chem-/sono-/photo-multimodal antitumor therapies has become an efficient strategy to combat malignant tumors. However, the existence of hypoxia in the tumor microenvironment (TME) leads to limited sonodynamic or photodynamic efficiency because O2 is the key reactant during the process of generation of reactive oxygen species (ROS). Here, to design a desirable platform that can simultaneously convert H2O2 in the TME into ROS and O2 for efficient chem-/sono-/photo-multimodal tumor therapies, we have created ultrasmall Cu2O-coordinated carbon nitride on a biocompatible ceria substrate (denoted as Cu2O-CNx@CeO2) via a self-assisted catalytic growth strategy. The chemical and morphological structures, ROS and O2 generation activities, and chemo-/photo-/sono-dynamic specificities of Cu2O-CNx@CeO2 when serving as multifunctional biocatalytic agents were systematically disclosed. The experimental studies validated that Cu2O-CNx@CeO2 presents state-of-the-art peroxidase-like and catalase-like activities. Moreover, the light excitation and ultrasound irradiation were also demonstrated to boost ROS production. The in vitro and in vivo experiments suggest that Cu2O-CNx@CeO2 can efficiently inhibit the growth of malignant melanoma via chem-/sono-/photo-multimodal antitumor ability. We believe that applying these new biocatalysts with dual catalytic activities of producing ROS and O2 will offer a new path for engineering multimodal nanoagents to combat malignant tumors.

A systematic observation of vasodynamics from different segments along the cerebral vasculature in the penumbra zone of awake mice following cerebral ischemia and recanalization.

Different segments of the cerebral vascular network may react distinctly to brain ischemia and recanalization. However, there are limited systematic observations of these vascular responses in mice under a physiological state following ischemic stroke. Herein, we aimed to investigate the vasodynamics among several segments along the cerebral vessels in awake mice following cerebral ischemia/recanalization via two-photon imaging. Plasma in the blood vessels were labelled with fluorescein isothiocyanate dextran. Smooth muscle cells and pericytes were labelled via a genetic mouse line (PDGFRß-tdTomato). We observed a no-reflow phenomenon in downstream microcirculation, and the vasodynamics of different segments of larger cerebral vessels varied in the penumbra area following cerebral ischemia-reperfusion. Despite obtaining reperfusion from the middle cerebral artery, there were significant constrictions of the downstream blood vessels in the ischemic penumbra zone. Interestingly, we observed an extensive constriction of the capillaries 3 hours following recanalization, both at the site covered by pericyte soma and by the pericyte process alone. In addition, we did not observe a significant positive correlation between the changed capillary diameter and pericyte coverage along the capillary. Taken together, abnormal constrictions and vasodynamics of cerebral large and small vessels may directly contribute to microcirculation failure following recanalization in ischemic stroke.

Ultrasound-derived muscle assessment system for older adults: a promising muscle mass estimation tool.

BACKGROUND: Quantitative assessment of muscle mass is a critical step in sarcopenia disease management. Expanding upon the use of ultrasound in foetal growth assessment, we established and validated an ultrasound-derived muscle assessment system for older adults at a risk of sarcopenia. METHODS: A total of 669 older adults were recruited in three cohorts in this cross-sectional study. In cohort 1(n = 103), the most valuable sites for skeletal muscle mass index (SMI) estimation were located among 11 ultrasound scanning sites. An ultrasound-derived SMI estimating algorithm based on muscle thickness (MT) was obtained in the modelling group composed of cohorts 1 and 2 (n = 309). The reliability of the muscle mass estimation equation and the validity of the obtained cut-off values were verified in cohort 3 (n = 257), which was selected as the verification group. RESULTS: In the modelling group, the cut-off values of ultrasound-derived e-SMI for low SMI were 7.13 kg/m2 for men and 5.81 kg/m2 for women. In the verification group, the intraclass correlation between e-SMI and SMI was 0.885. The sensitivity of the e-SMI in detecting low SMI was 93.6% for men and 89.7% for women, and the negative predictive value was 94.9% for men and 94.7% for women. Combined with the handgrip strength and gait speed, the e-SMI had an overall diagnostic sensitivity of 92.7% and a specificity of 91.0% for sarcopenia. CONCLUSION: The ultrasound-derived muscle assessment system can be a promising muscle mass estimation tool and a potential disease classification tool.

e-TSN: an interactive visual exploration platform for target-disease knowledge mapping from literature.

Target discovery and identification processes are driven by the increasing amount of biomedical data. The vast numbers of unstructured texts of biomedical publications provide a rich source of knowledge for drug target discovery research and demand the development of specific algorithms or tools to facilitate finding disease genes and proteins. Text mining is a method that can automatically mine helpful information related to drug target discovery from massive biomedical literature. However, there is a substantial lag between biomedical publications and the subsequent abstraction of information extracted by text mining to databases. The knowledge graph is introduced to integrate heterogeneous biomedical data. Here, we describe e-TSN (Target significance and novelty explorer, http://www.lilab-ecust.cn/etsn/), a knowledge visualization web server integrating the largest database of associations between targets and diseases from the full scientific literature by constructing significance and novelty scoring methods based on bibliometric statistics. The platform aims to visualize target-disease knowledge graphs to assist in prioritizing candidate disease-related proteins. Approved drugs and associated bioactivities for each interested target are also provided to facilitate the visualization of drug-target relationships. In summary, e-TSN is a fast and customizable visualization resource for investigating and analyzing the intricate target-disease networks, which could help researchers understand the mechanisms underlying complex disease phenotypes and improve the drug discovery and development efficiency, especially for the unexpected outbreak of infectious disease pandemics like COVID-19.

VRPharmer: bringing virtual reality into pharmacophore-based virtual screening with interactive exploration and realistic visualization.

SUMMARY: Current pharmacophore-based virtual screening (VS) software has limited interactive capabilities and less intuitive screening processes. In this study, a novel tool named VRPharmer is proposed to perform the entire VS workflow in VR environments. VRPharmer enables users to interactively perceive computation processes and immersively observe molecular structures. Besides a typical screening mode (OPT mode), VRPharmer provides a unique interactive screening mode (SCORE mode) for freely exploring the optimal binding poses. Pharmacophore models are editable to study the impact of each feature and further refine the screening results. Moreover, molecular rendering algorithms are improved for precise representations. AVAILABILITY AND IMPLEMENTATION: VRPharmer is open-source software under the MIT license. The released version is available at https://github.com/VRPharmer/VRPharmer. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

A Broad Antiviral Strategy: Inhibitors of Human DHODH Pave the Way for Host-Targeting Antivirals against Emerging and Re-Emerging Viruses.

New strategies to rapidly develop broad-spectrum antiviral therapies are urgently required for emerging and re-emerging viruses. Host-targeting antivirals (HTAs) that target the universal host factors necessary for viral replication are the most promising approach, with broad-spectrum, foresighted function, and low resistance. We and others recently identified that host dihydroorotate dehydrogenase (DHODH) is one of the universal host factors essential for the replication of many acute-infectious viruses. DHODH is a rate-limiting enzyme catalyzing the fourth step in de novo pyrimidine synthesis. Therefore, it has also been developed as a therapeutic target for many diseases relying on cellular pyrimidine resources, such as cancers, autoimmune diseases, and viral or bacterial infections. Significantly, the successful use of DHODH inhibitors (DHODHi) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection further supports the application prospects. This review focuses on the advantages of HTAs and the antiviral effects of DHODHi with clinical applications. The multiple functions of DHODHi in inhibiting viral replication, stimulating ISGs expression, and suppressing cytokine storms make DHODHi a potent strategy against viral infection.

SNHG1/miR-145-5p/KLF5 Axis Participates in Regulating the Proliferation and Migration of Oral Squamous Cell Cancer.

We aimed to clarify the molecular mechanism of lncRNA SNHG1 in regulating the OSCC process. Clinical samples of OSCC were collected for detecting the differential level of SNHG1 by qRT-PCR. Pathological indexes of OSCC patients were analyzed for uncovering the prognostic value of SNHG1. The interaction between SNHG1 and miR-145-5p was assessed through the bioinformatics method and dual-luciferase reporter assay. Their coregulation on proliferative and migratory functions of Tca8113 and CAL-27 cells was explored by the CCK-8, EdU, and Transwell assay. Finally, the regulatory effect of miR-145-5p on its downstream gene KLF5 was evaluated. SNHG1 was abnormally upregulated in OSCC samples and linked to a poor prognosis of OSCC patients. Serving as an oncogene, SNHG1 strengthened proliferative and migratory functions of Tca8113 and CAL-27 cells. miR-145-5p was a key downstream target inducing the oncogenic role of SNHG1 in the OSCC process with KLF5 as its downstream gene. SNHG1/miR-145-5p/KLF1 axis is responsible for driving the malignant process of OSCC.

Homology and Immune Checkpoint Dual-Targeted Sonocatalytic Nanoagents for Enhancing Sonodynamic Tumor Therapy.

Sonocatalytic nanoagents (SCNs), a kind of sonosensitizers, could catalyze oxygen to generate abundant reactive oxygen species (ROS) under stimulations of noninvasive and deep-penetrating ultrasound (US), which is commonly used for sonodynamic therapy (SDT) of tumors such as malignant melanoma. However, poor bioavailability of most SCNs and fast quenching of extracellular-generating ROS from SDT limit further applications of SCNs in the SDT of tumors. Herein, we synthesized a new kind of TiO2-based SCN functionalized with the malignant melanoma cell membrane (B16F10M) and programmed cell death-ligand 1 antibody (aPD-L1) for homology and immune checkpoint dual-targeted and enhanced sonodynamic tumor therapy. Under US irradiation, the synthesized SCN can catalytically generate a large amount of 1O2. In vitro experiments validate that functionalized SCNs exhibit precise targeting effects, high tumor cell uptake, and intracellular sonocatalytic killing of the B16F10 cells by a large amount of localized ROS. Utilizing the melanoma animal model, the functionalized SCN displays visible long-term retention in the tumor area, which assists the homology and immune checkpoint synergistically dual-targeted and enhanced in vivo SDT of the tumor. We suggest that this highly bioavailable and dual-functionalized SCN may provide a promising strategy and nanoplatform for enhancing sonodynamic tumor therapies.

Mechanism of persulfate activation by biochar for the catalytic degradation of antibiotics: Synergistic effects of environmentally persistent free radicals and the defective structure of biochar.

The abuse of antibiotics threatens the water environment and human health. Green treatment method is needed to degrade antibiotics such as biochar. Few studies have examined the environmentally persistent free radicals (EPFRs) and defective structure of biochar during the biochar-mediated catalytic degradation of antibiotics. In this study, biochar prepared from poplar and pine sawdust was used to activate peroxymonosulfate (PMS) to generate instant radicals (SO4•- and •OH) and degrade tetracycline (TC), chlortetracycline (CTC) and doxycycline (DOX). The preparation temperatures ranged from 300 °C to 900 °C. EPFRs were the main activator of PMS at 300-500 °C, and the defective structure of biochar was the main activator at 800-900 °C. The concentrations of EPFRs ranged from 1.75 × 1018 spins/g to 6.44 × 1018 spins/g. According to the electron paramagnetic resonance (EPR) parameter (g-factor), the main types of EPFRs were carbon-centered radicals (g1 < 2.0030) or carbon-centered radicals with oxygen atoms (2.0030 < g2 < 2.0040). Optimization of the degradation experiment revealed that the removal rate of antibiotics peaked when the preparation temperature was 500 °C and 900 °C. In the biochar/PMS system, the antibiotics removal rate of 90% was achieved in 40 min with an average apparent rate constant (kobs) of 0.0588 min-1. Analysis of the mechanism revealed that the free radical pathway (EPFRs and defective structure) can effectively activate PMS to generate SO4•- and •OH. However, control experiments suggested that the non-free radical pathway (singlet oxygen) had little effect on antibiotic degradation. After five cycles, the removal rate of antibiotics by biochar was still greater than 70%, indicating that biochar retains a high degradation ability. These results indicate that optimizing the preparation conditions can effectively expand the application range of the biochar/PMS system and improve the degradation of antibiotic wastewater.

Efficacy of Extracorporeal Shock Wave Therapy for Achilles Tendinopathy: A Meta-analysis.

BACKGROUND: Achilles tendinopathy is a frequent sports injury, and extracorporeal shock wave therapy (ESWT) has been proposed as a treatment. PURPOSE: To compare outcomes between ESWT and other nonsurgical intervention (including sham shock wave therapy) in Achilles tendinopathy patients. STUDY DESIGN: Systematic review; Level of evidence, 2. METHODS: We included 5 randomized controlled trials and 3 case-control studies published between 2005 and 2018. We analyzed pain scores and other outcomes that were reported in more than 3 of the 8 studies. RESULTS: ESWT was associated with significantly better scores than comparison therapy on the visual analog scale for pain (P < .01), American Orthopaedic Foot & Ankle Society scale (P = .01), Likert scale for satisfaction (P = .03), Roles and Maudsley scale (P < .01), Victorian Institute of Sports Assessment-Achilles questionnaire (P < .01), and numerical rating scale (P = .02). The 2 patient groups did not differ significantly in tenderness (P = .34) or pain threshold (P = .24). Subgroup analysis showed that ESWT led to better VAS pain scores than comparison treatments at both low-energy level (0.06-0.11 mJ/mm2) and medium-energy level (0.12-0.25 mJ/mm2) and at both shorter (<6 months) and longer (≥6 months) follow-up. CONCLUSION: ESWT improves pain and functional outcomes in patients with Achilles tendinopathy. Further research is needed to determine the optimal energy level.

Nitro-oleic acid inhibits the high glucose-induced epithelial-mesenchymal transition in peritoneal mesothelial cells and attenuates peritoneal fibrosis.

As an electrophilic nitroalkene fatty acid, nitro-oleic acid (OA-NO2) exerts multiple biological effects that contribute to anti-inflammation, anti-oxidative stress, and antiapoptosis. However, little is known about the role of OA-NO2 in peritoneal fibrosis. Thus, in the present study, we examined the effects of OA-NO2 on the high glucose (HG)-induced epithelial-mesenchymal transition (EMT) in human peritoneal mesothelial cells (HPMCs) and evaluated the morphological and immunohistochemical changes in a rat model of peritoneal dialysis-related peritoneal fibrosis. In in vitro experiments, we found that HG reduced the expression level of E-cadherin and increased Snail, N-cadherin, and α-smooth muscle actin expression levels in HPMCs. The above-mentioned changes were attenuated by pretreatment with OA-NO2. Additionally, OA-NO2 also inhibited HG-induced activation of the transforming growth factor-ß1/Smad signaling pathway and NF-κB signaling pathway. Meanwhile, OA-NO2 inhibited HG-induced phosphorylation of Erk and JNK. The results from the in vivo experiments showed that OA-NO2 notably relieved peritoneal fibrosis by decreasing the thickness of the peritoneum; it also inhibited expression of transforming growth factor-ß1, α-smooth muscle actin, N-cadherin, and vimentin and enhanced expression of E-cadherin in the peritoneum. Collectively, these results suggest that OA-NO2 inhibits the HG-induced epithelial-mesenchymal transition in HPMCs and attenuates peritoneal dialysis-related peritoneal fibrosis.

[Research progress of scaffold materials for tissue engineered meniscus].

OBJECTIVE: To summarize and analyze the research progress of scaffold materials used in tissue engineered meniscus. METHODS: The classification and bionics design of scaffold materials were summarized by consulting domestic and foreign literature related to the research of tissue engineered meniscus in recent years. RESULTS: Tissue engineered meniscus scaffolds can be roughly classified into synthetic polymers, hydrogels, extracellular matrix components, and tissue derived materials. These different materials have different characteristics, so the use of a single material has its unique disadvantages, and the use of a variety of materials composite scaffolds can learn from each other, which is a hot research area at present. In addition to material selection, material processing methods are also the focus of research. At the same time, according to the morphological structure and mechanical characteristics of the meniscus, the bionic design of tissue engineered meniscus scaffolds has great potential. CONCLUSION: At present, there are many kinds of scaffold materials for tissue engineered meniscus. However, there is no material that can completely simulate the natural meniscus, and further research of scaffold materials is still needed.

Relation of the pdxB-usg-truA-dedA Operon and the truA Gene to the Intracellular Survival of Salmonella enterica Serovar Typhimurium.

Salmonella is the genus of Gram-negative, facultative intracellular pathogens that have the ability to infect large numbers of animal or human hosts. The S. enterica usg gene is associated with intracellular survival based on ortholog screening and identification. In this study, the λ-Red recombination system was used to construct gene deletion strains and to investigate whether the identified operon was related to intracellular survival. The pdxB-usg-truA-dedA operon enhanced the intracellular survival of S. enterica by resisting the oxidative environment and the usg and truA gene expression was induced by H2O2. Moreover, the genes in this operon (except for dedA) contributed to virulence in mice. These findings indicate that the pdxB-usg-truA-dedA operon functions in resistance to oxidative environments during intracellular survival and is required for in vivo S. enterica virulence. This study provides insight toward a better understand of the characteristics of intracellular pathogens and explores the gene modules involved in their intracellular survival.