Mn2+ recycling in hypersaline wastewater: unnoticed intracellular biomineralization and pre-cultivation of immobilized bacteria.

Microbially induced manganese carbonate precipitation has been utilized for the treatment of wastewater containing manganese. In this study, Virgibacillus dokdonensis was used to remove manganese ions from an environment containing 5% NaCl. The results showed a significant decrease in carbonic anhydrase activity and concentrations of carbonate and bicarbonate ions with increasing manganese ion concentrations. However, the levels of humic acid analogues, polysaccharides, proteins, and DNA in EPS were significantly elevated compared to those in a manganese-free environment. The rhodochrosite exhibited a preferred growth orientation, abundant morphological features, organic elements including nitrogen, phosphorus, and sulfur, diverse protein secondary structures, as well as stable carbon isotopes displaying a stronger negative bias. The presence of manganese ions was found to enhance the levels of chemical bonds O-C=O and N-C=O in rhodochrosite. Additionally, manganese in rhodochrosite exhibited both + 2 and + 3 valence states. Rhodochrosite forms not only on the cell surface but also intracellularly. After being treated with free bacteria for 20 days, the removal efficiency of manganese ions ranged from 88.4 to 93.2%, and reached a remarkable 100% on the 10th day when using bacteria immobilized on activated carbon fiber that had been pre-cultured for three days. The removal efficiency of manganese ions was significantly enhanced under the action of pre-cultured immobilized bacteria compared to non-pre-cultured immobilized bacteria. This study contributes to a comprehensive understanding of the mineralization mechanism of rhodochrosite, thereby providing an economically and environmentally sustainable biological approach for treating wastewater containing manganese.

Screening a DNA Aptamer Specifically Targeting Integrin ß3 and Partially Inhibiting Tumor Cell Migration.

Due to its key roles in malignant tumor progression and reprograming of the tumor microenvironment, integrin ß3 has attracted great attention as a new target for tumor therapy. However, the structure-function relationship of integrins ß3 remains incompletely understood, leading to the shortage of specific and effective targeting probes. This work uses a purified extracellular domain of integrin ß3 and integrin ß3-positive cells to screen aptamers, specifically targeting integrin ß3 in the native conformation on live cells through the SELEX approach. Following meticulous truncation and characterization of the initial aptamer candidates, the optimized aptamer S10yh2 was produced, exhibiting a low equilibrium dissociation constant (Kd) in the nanomolar range. S10yh2 displays specific recognition of cancer cells with varying levels of integrin ß3 expression and demonstrates favorable stability in serum. Subsequent analysis of docking sites revealed that S10yh2 binds to the seven amino acid residues located in the core region of integrin ß3. The S10yh2 aptamer can downregulate the level of integrin heterodimer αvß3 on integrin ß3 overexpressed cancer cells and partially inhibit cell migration behavior. In summary, S10yh2 is a promising probe with a small size, simple synthesis, good stability, high binding affinity, and selectivity. It therefore holds great potential for investigating the structure-function relationship of integrins.

The mediating role of psychological capital between post-traumatic growth and uncertainty in illness among patients with Parkinson's disease.

With the application of positive psychology in health management, many studies have confirmed the close relationship between post-traumatic growth, psychological capital and uncertainty in illness. However, there is still a lack of attention to the positive psychology of patients with Parkinson's disease, and previous studies have not clarify the specific relationship. Therefore, the present study sought to explore the multiple mediating roles of psychological capital between post-traumatic growth and uncertainty in illness among patients with Parkinson's disease. We conducted a cross-sectional survey, a total of 268 patients with Parkinson's disease were investigated by the Post-Traumatic Growth Inventory, the Positive Psychological Capital Questionnaire and the Mishel Uncertainty in Illness Scale - Adults from November 2021 to June 2022 in the Parkinson's specialist outpatient department of three 3a-grade hospitals in Tianjin, China. The results showed that the score of post-traumatic growth was (51.78±17.872), the score of uncertainty in illness was (68.08±17.555), and the score of self-efficacy, resilience, hope, and optimism dimensions of psychological capital were (28.93±10.511), (28.79±11.553), (27.57±8.558) and (25.35±9.768). Post-traumatic growth was negatively correlated with uncertainty in illness and positively correlated with all four dimensions of psychological capital (p < 0.01), and uncertainty in illness was negatively correlated with all four dimensions of psychological capital (p < 0.01). Bootstrap test showed that the direct effect of post-traumatic growth on uncertainty in illness of patients with Parkinson's disease was not significant; the total indirect effect of psychological capital between post-traumatic growth and uncertainty in illness was significant. The mediating effects of resilience and hope were significant, accounting for 53.07% and 19.41% of the total indirect effects, respectively. Based on the research results, psychological capital played a completely mediating role between post-traumatic growth and uncertainty in illness of patients with Parkinson's disease. Post-traumatic growth affected uncertainty in illness through two dimensions: resilience and hope. Healthcare professionals should pay attention to the level of psychological capital of patients with Parkinson's disease and implement targeted positive psychological interventions focusing on the two dimensions of resilience and hope, to enhance post-traumatic growth and reduce the level of uncertainty in illness.

An Activity-Based Sensing Fluorogenic Probe for Monitoring Ethylene in Living Cells and Plants.

Ethylene (ET) is an important gaseous plant hormone. It is highly desirable to develop fluorescent probes for monitoring ethylene in living cells. We report an efficient RhIII -catalysed coupling of N-phenoxyacetamides to ethylene in the presence of an alcohol. The newly discovered coupling reaction exhibited a wide scope of N-phenoxyacetamides and excellent regioselectivity. We successfully developed three fluorophore-tagged RhIII -based fluorogenic coumarin-ethylene probes (CEPs) using this strategy for the selective and quantitative detection of ethylene. CEP-1 exhibited the highest sensitivity with a limit of detection of ethylene at 52 ppb in air. Furthermore, CEP-1 was successfully applied for imaging in living CHO-K1 cells and for monitoring endogenous-induced changes in ethylene biosynthesis in tobacco and Arabidopsis thaliana plants. These results indicate that CEP-1 has great potential to illuminate the spatiotemporal regulation of ethylene biosynthesis and ethylene signal transduction in living biological systems.

Establishment of a serological molecular model for the early diagnosis and progression monitoring of bone metastasis in lung cancer.

BACKGROUND: The prognosis is very poor for lung cancer patients with bone metastasis. Unfortunately, a suitable method has yet to become available for the early diagnosis of bone metastasis in lung cancer patients. The present work describes an attempt to develop a novel model for the early identification of lung cancer patients with bone metastasis risk. METHODS: As the test group, 205 primary lung cancer patients were recruited, of which 127 patients had bone metastasis; the other 78 patients without bone metastasis were set as the negative control. Additionally, 106 healthy volunteers were enrolled as the normal control. Serum levels of several cytokines in the bone microenvironment (CaN, OPG, PTHrP, and IL-6) and bone turnover markers (tP1NP, ß-CTx) were detected in all samples by ECLIA or ELISA assay. Receiver operating characteristic (ROC) curves and multivariate analyses were performed to evaluate diagnostic abilities and to assess the attributable risk of bone metastasis for each of these indicators; the diagnostic model was established via logistic regression analysis. The prospective validation group consisted of 44 patients with stage IV primary lung cancer on whom a follow-up of at least 2 years was conducted, during which serum bone biochemical marker concentrations were monitored. RESULTS: The serological molecular model for the diagnosis of bone metastasis was logit (p). ROC analysis showed that when logit (p) > 0.452, the area under curve of the model was 0.939 (sensitivity: 85.8%, specificity: 89.7%). Model validation demonstrated accuracy with a high degree of consistency (specificity: 85.7%, specificity: 87.5%, Kappa: 0.770). The average predictive time for bone metastasis occurrence of the model was 9.46 months earlier than that of the bone scan diagnosis. Serum OPG, PTHrP, tP1NP, ß-CTx, and the diagnostic model logit (p) were all positively correlated with bone metastasis progression (P < 0.05). CONCLUSIONS: This diagnostic model has the potential to be a simple, non-invasive, and sensitive tool for diagnosing the occurrence and monitoring the progression of bone metastasis in patients with lung cancer.

Pseudoesotropia in Chinese Children: A Triphasic Development of the Interepicanthal Folds Distance-to-Interpupillary Distance Ratio and Its Changing Perception.

PURPOSE: To delineate the development of the interepicanthal fold distance (IEFD) to interpupillary distance (IPD) in Chinese children, and to quantify how their ratio (EFDPD ratio) affects parent's judgment on whether a child's two eyes appear misaligned. METHODS: The values of IPD and IEFD were measured in 750 children, aged between 3 and 17 years. The developmental trend of EFDPD ratio was established. Two hundred parents were shown a series of pictures of children with varying EFDPD ratios and asked to judge whether the child in each picture demonstrated misaligned eyes. Based on the parent's responses, psychometric functional associations with EFDPD ratios were established. RESULTS: The EFDPD ratios were significantly higher (0.63 ± 0.027) and showed little change among children from 3 to 6 years of age (p = 0.704). During the age of seven to 12 years, however, the EFDPD ratio significantly decreased (p < 0.001) before stabilizing at 0.59 ± 0.023 by the ages of 13 to 17 years (p = 0.376). Children with EFDPD ratios > 0.65 were more likely to be perceived as strabismic by the parents, while children with an EFDPD ratio < 0.55 were rarely perceived as so. As many as 30% of the children aged between 3 and 6 years demonstrated EFDPD ratios > 0.65, and this number reduced to 5% by the age of 12 years. CONCLUSIONS: The development of the EFDPD ratio in Chinese children shows a triphasic pattern, with a large value before the age of 6 years, a quick drop between 7 and 12 years, and little change after 13 years of age. As the EFDPD ratio declines, fewer children appear as strabismic. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Overlapping hotspots in CDRs are critical sites for V region diversification.

Activation-induced deaminase (AID) mediates the somatic hypermutation (SHM) of Ig variable (V) regions that is required for the affinity maturation of the antibody response. An intensive analysis of a published database of somatic hypermutations that arose in the IGHV3-23*01 human V region expressed in vivo by human memory B cells revealed that the focus of mutations in complementary determining region (CDR)1 and CDR2 coincided with a combination of overlapping AGCT hotspots, the absence of AID cold spots, and an abundance of polymerase eta hotspots. If the overlapping hotspots in the CDR1 or CDR2 did not undergo mutation, the frequency of mutations throughout the V region was reduced. To model this result, we examined the mutation of the human IGHV3-23*01 biochemically and in the endogenous heavy chain locus of Ramos B cells. Deep sequencing revealed that IGHV3-23*01 in Ramos cells accumulates AID-induced mutations primarily in the AGCT in CDR2, which was also the most frequent site of mutation in vivo. Replacing the overlapping hotspots in CDR1 and CDR2 with neutral or cold motifs resulted in a reduction in mutations within the modified motifs and, to some degree, throughout the V region. In addition, some of the overlapping hotspots in the CDRs were at sites in which replacement mutations could change the structure of the CDR loops. Our analysis suggests that the local sequence environment of the V region, and especially of the CDR1 and CDR2, is highly evolved to recruit mutations to key residues in the CDRs of the IgV region.

The efficacious combined treatment of rhegmatogenous retinal detachment (PVR ≤ C2) with inferior breaks using 25-gauge pars plana vitrectomy and air tamponade.

To examine the surgical results of 25-gauge (25-G) pars plana vitrectomy (PPV) with air tamponade for the repair of primary rhegmatogenous retinal detachment (RRD) with inferior retinal breaks. A retrospective consecutive study was conducted from March 2019 to February 2023 on 27 patients with primary RRD with inferior breaks who underwent 25-G PPV with air tamponade. Once the surgery was completed, the patients were monitored for a minimum of 6 months. Postoperative best-corrected visual acuity (BCVA), postoperative complications, and the rate of single surgery anatomical success were the primary outcome measures. The average age of the 12 female and 15 male study participants was 50.2 ±â€…12.8 years. The follow-up period was 10.3 ±â€…3.9 months on average. On average, the affected clock hours were 5.6 ±â€…2.1 hours, and the average number of retinal breaks was 2.2 ±â€…1.8. Moreover, the final anatomical success rate was 100%, compared to the estimated initial anatomical success rate of 96.30% following a single surgery. At 6 months, we also noticed a significant change in the mean BCVA, which went from 1.62 ±â€…0.71 logMAR (preoperative) to 0.89 ±â€…0.61 logMAR (postoperative) (P < .001). On the first day and the week following surgery, the mean intraocular pressure (IOP) was similar (all P > .05). One patient experienced postoperative complications such as retinal redetachment, which was addressed with the same procedure. After surgery, 2 patients had IOP of 35 and 28 mm Hg for 1 week. Two weeks following surgery, their IOP essentially returned to normal after receiving antihypertensive medication. No other notable postoperative complications were observed. According to this research, 25-G PPV with air tamponade has a satisfactory success rate in repairing primary RRD with inferior breaks. This procedure also produces quicker visual recovery and is linked to fewer complications.

The effects of Baduanjin on physical function, daily living activities and quality of life in patients with stroke: a systematic review and meta-analysis.

OBJECTIVES: Baduanjin is a traditional Chinese exercise that combines respiration and movement. The study is designed to evaluate the effects of Baduanjin on physical function, daily living activities, and quality of life in stroke patients. METHODS: Eleven databases were searched from inception through 16 September 2022. The methodological quality evaluation of the studies was evaluated with the Cochrane Handbook for Systematic Reviews of Interventions v5.1.0. The quality evaluation of the evidence was graded with the Guideline Development Tool . RESULTS: A total of 24 studies were included. Three indicators with moderate quality evidence, six indicators with low quality evidence, and one indicator with very low quality evidence. Meta-analysis results suggest that Baduanjin group is better than the control group in increasing Berg balance scale [MD = 7.99, 95%CI (4.99, 10.99), P < 0.00001], Fugl- Meyer balance scale [MD = 5.26, 95%CI (0.12, 10.40), P = 0.04]; Total Fugl-Meyer assessment scale [MD = 7.17, 95%CI (3.13, 11.2), P = 0.0005], Fugl-Meyer assessment lower extremity scale[MD = 3.05, 95%CI(0.95, 5.15), P = 0.004], Fugl-Meyer assessment upper extremity scale[MD = 4.60, 95%CI (1.64,7.56), P = 0.002]; walking function scale[SMD = 0.69, 95%CI (0.14, 1.23), P = 0.01]; trunk function scale[MD = 2.46, 95%CI (1.75,3.17), P < 0.00001]; Barthel index[MD = 9.43, 95%CI (6.20,12.65), P < 0.00001] and quality of life score[SMD = 1.85, 95%CI (1.46, 2.23), P < 0.00001]. Meanwhile, Baduanjin reduced Neurological deficit scores SMD = -1.41, 95%CI (-1.83, -0.98), P < 0.00001]). CONCLUSIONS: Baduanjin is probably an effective rehabilitation method to improve balance, motor, trunk, neurological functions, daily living activities, and quality of life in stroke patients. However, the result of Baduanjin in improving walking function is unstable, and further high-quality randomized controlled trials are needed to confirm the results.

The impact of empowerment theory-based health education on Alzheimer's disease informal caregivers: a randomized controlled trial.

Background: There is a lack of evidence regarding the effectiveness of empowerment healthy education for caregivers of Alzheimer's patients. Objective: To explore the effectiveness of the intervention of health education guided by empowerment theory on dementia knowledge, caregiving readiness, positive caregiving emotions, anxiety, and depression in informal Alzheimer's disease caregivers. Design: A single-blinded, randomized controlled trial. Setting: A teaching hospital in Tianjin, China. Participants: Eighty caregivers of Alzheimer's disease patients. Methods: Participants were recruited from the hospital and randomly assigned to either experimental or control group. The experimental group underwent a 12-weeks, one-to-one intervention of six session lasting 45-60 min each. The control group received conventional health education. Outcome measures included dementia knowledge, caregiver readiness (primary outcomes), positive caregiving emotions, anxiety, and depression (secondary outcomes). Results: After 12 weeks, the intervention group exhibited significantly higher levels of dementia knowledge, caregiver readiness, and positive caregiving emotions compared with the control group. Furthermore, levels of hospitalization-related anxiety and depression were lower in the intervention group. All study results of this study showed statistically significant differences (p < 0.05). Discussion: Empowerment theory-based health education appears to be an effective intervention for improving caregiver and readiness to care for caregivers of Alzheimer's disease individuals. The intervention may help reduce caregivers' anxiety and depression levels.

Development and validation of an early diagnosis model for bone metastasis in non-small cell lung cancer based on serological characteristics of the bone metastasis mechanism.

Background: Bone metastasis significantly impact the prognosis of non-small cell lung cancer (NSCLC) patients, reducing their quality of life and shortening their survival. Currently, there are no effective tools for the diagnosis and risk assessment of early bone metastasis in NSCLC patients. This study employed machine learning to analyze serum indicators that are closely associated with bone metastasis, aiming to construct a model for the timely detection and prognostic evaluation of bone metastasis in NSCLC patients. Methods: The derivation cohort consisted of 664 individuals with stage IV NSCLC, diagnosed between 2015 and 2018. The variables considered in this study included age, sex, and 18 specific serum indicators that have been linked to the occurrence of bone metastasis in NSCLC. Variable selection used multivariate logistic regression analysis and Lasso regression analysis. Six machine learning methods were utilized to develop a bone metastasis diagnostic model, assessed with Area Under the Curve (AUC), Decision Curve Analysis (DCA), sensitivity, specificity, and validation cohorts. External validation used 113 NSCLC patients from the Medical Alliance (2019-2020). Furthermore, a prospective validation study was conducted on a cohort of 316 patients (2019-2020) who were devoid of bone metastasis, and followed-up for at least two years to assess the predictive capabilities of this model. The model's prognostic value was evaluated using Kaplan-Meier survival curves. Findings: Through variable selection, 11 serum indictors were identified as independent predictive factors for NSCLC bone metastasis. Six machine learning models were developed using age, sex, and these serum indicators. A random forest (RF) model demonstrated strong performance during the training and internal validation cohorts, achieving an AUC of 0.98 (95% CI 0.95-0.99) for internal validation. External validation further confirmed the RF model's effectiveness, yielding an AUC of 0.97 (95% CI 0.94-0.99). The calibration curves demonstrated a high level of concordance between the anticipated risk and the observed risk of the RF model. Prospective validation revealed that the RF model could predict the occurrence of bone metastasis approximately 10.27 ± 3.58 months in advance, according to the results of the SPECT. An online computing platform (https://bonemetastasis.shinyapps.io/shiny_cls_1model/) for this RF model is publicly available and free-to-use by doctors and patients. Interpretation: This study innovatively employs age, gender, and 11 serological markers closely related to the mechanism of bone metastasis to construct an RF model, providing a reliable tool for the early screening and prognostic assessment of bone metastasis in NSCLC patients. However, as an exploratory study, the findings require further validation through large-scale, multicenter prospective studies. Funding: This work is supported by the National Natural Science Foundation of China (NO.81974315); Shanghai Municipal Science and Technology Commission Medical Innovation Research Project (NO.20Y11903300); Shanghai Municipal Health Commission Health Industry Clinical Research Youth Program (NO.20204Y034).

The Performance of Spot Photoscreener in 6 to 10 Weeks Infants in China: A Cross-Sectional Study.

Purpose: To compare the refractive errors measured by the Spot photoscreener (with or without cycloplegia) to cycloplegic retinoscopy in 6- to 10-week-old infants. Materials and Methods: 101 right eyes from 101 healthy infants aged 6 to 10 weeks were recruited for this cross-sectional observational study. Refractive errors were measured using Spot photoscreener before and after cycloplegia, as well as cycloplegic retinoscopy. Comparisons between the refractive measurements were performed using one-way ANOVA with the post hoc Tukey HSD test or Kruskal-Wallis test with the Steel-Dwass test according to the data normality. Pearson's correlation test and 95% confidence intervals were calculated. The agreement was evaluated using a Bland-Altman plot with 95% limits of agreement of the differences. Results: Spot photoscreener was found to underestimate the spherical equivalent by 2.33 Diopters (D) in these infants. Following the induction of cycloplegia, the spherical equivalent measured by Spot photoscreener was in excellent agreement with cycloplegic retinoscopy with the mean difference of 0.01 D. Spot photoscreener overestimated cylindrical parameter by 0.2 D with poor agreement with cycloplegic retinoscopy no matter whether cycloplegia was induced. It had good agreement with cycloplegic retinoscopy in the J0 vector than the J45 vector measurement. Conclusions: With the induction of cycloplegia, Spot photoscreener can accurately evaluate spherical equivalent in hyperopic infants with mild-to-moderate astigmatism. While it may provide valuable measurements of astigmatism, discrepancies in cylinder and axis should be taken into account.

Bacillus cereus NJ01 induces SA- and ABA-mediated immunity against bacterial pathogens through the EDS1-WRKY18 module.

Emerging evidence suggests a beneficial role of rhizobacteria in ameliorating plant disease resistance in an environment-friendly way. In this study, we characterize a rhizobacterium, Bacillus cereus NJ01, that enhances bacterial pathogen resistance in rice and Arabidopsis. Transcriptome analyses show that root inoculation of NJ01 induces the expression of salicylic acid (SA)- and abscisic acid (ABA)-related genes in Arabidopsis leaves. Genetic evidence showed that EDS1, PAD4, and WRKY18 are required for B. cereus NJ01-induced bacterial resistance. An EDS1-PAD4 complex interacts with WRKY18 and enhances its DNA binding activity. WRKY18 directly binds to the W box in the promoter region of the SA biosynthesis gene ICS1 and ABA biosynthesis genes NCED3 and NCED5 and contributes to the NJ01-induced bacterial resistance. Taken together, our findings indicate a role of the EDS1/PAD4-WRKY18 complex in rhizobacteria-induced disease resistance.

Arabidopsis Tyrosylprotein sulfotransferase acts in the auxin/PLETHORA pathway in regulating postembryonic maintenance of the root stem cell niche.

Recent identification of the Arabidopsis thaliana tyrosylprotein sulfotransferase (TPST) and a group of Tyr-sulfated peptides known as root meristem growth factors (RGFs) highlights the importance of protein Tyr sulfation in plant growth and development. Here, we report the action mechanism of TPST in maintenance of the root stem cell niche, which in the Arabidopsis root meristem is an area of four mitotically inactive quiescent cells plus the surrounding mitotically active stem cells. Mutation of TPST leads to defective maintenance of the root stem cell niche, decreased meristematic activity, and stunted root growth. We show that TPST expression is positively regulated by auxin and that mutation of this gene affects auxin distribution by reducing local expression levels of several PIN genes and auxin biosynthetic genes in the stem cell niche region. We also show that mutation of TPST impairs basal- and auxin-induced expression of the PLETHORA (PLT) stem cell transcription factor genes and that overexpression of PLT2 rescues the root meristem defects of the loss-of-function mutant of TPST. Together, these results support that TPST acts to maintain root stem cell niche by regulating basal- and auxin-induced expression of PLT1 and PLT2. TPST-dependent sulfation of RGFs provides a link between auxin and PLTs in regulating root stem cell niche maintenance.

A Proteomics Insight into Advancements in the Rice-Microbe Interaction.

Rice is one of the most-consumed foods worldwide. However, the productivity and quality of rice grains are severely constrained by pathogenic microbes. Over the last few decades, proteomics tools have been applied to investigate the protein level changes during rice-microbe interactions, leading to the identification of several proteins involved in disease resistance. Plants have developed a multi-layered immune system to suppress the invasion and infection of pathogens. Therefore, targeting the proteins and pathways associated with the host's innate immune response is an efficient strategy for developing stress-resistant crops. In this review, we discuss the progress made thus far with respect to rice-microbe interactions from side views of the proteome. Genetic evidence associated with pathogen-resistance-related proteins is also presented, and challenges and future perspectives are highlighted in order to understand the complexity of rice-microbe interactions and to develop disease-resistant crops in the future.

Suppression of ETI by PTI priming to balance plant growth and defense through an MPK3/MPK6-WRKYs-PP2Cs module.

Pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) are required for host defense against pathogens. Although PTI and ETI are intimately connected, the underlying molecular mechanisms remain elusive. In this study, we demonstrate that flg22 priming attenuates Pseudomonas syringae pv. tomato DC3000 (Pst) AvrRpt2-induced hypersensitive cell death, resistance, and biomass reduction in Arabidopsis. Mitogen-activated protein kinases (MAPKs) are key signaling regulators of PTI and ETI. The absence of MPK3 and MPK6 significantly reduces pre-PTI-mediated ETI suppression (PES). We found that MPK3/MPK6 interact with and phosphorylate the downstream transcription factor WRKY18, which regulates the expression of AP2C1 and PP2C5, two genes encoding protein phosphatases. Furthermore, we observed that the PTI-suppressed ETI-triggered cell death, MAPK activation, and growth retardation are significantly attenuated in wrky18/40/60 and ap2c1 pp2c5 mutants. Taken together, our results suggest that the MPK3/MPK6-WRKYs-PP2Cs module underlies PES and is essential for the maintenance of plant fitness during ETI.

Dynamic Changes of ORF1ab and N Gene Ct Values in COVID-19 Omicron Inpatients of Different Age Groups - Beijing Municipality, China, November-December 2022.

Introduction: In November 2021, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant was identified as the variant of concern and has since spread globally, replacing other cocirculating variants. To better understand the dynamic changes in viral load over time and the natural history of the virus infection, we analyzed the expression of the open reading frames 1ab (ORF1ab) and nucleocapsid (N) genes in patients infected with Omicron. Methods: We included patients initially admitted to the hospital for SARS-CoV-2 infection between November 5 and December 25, 2022. We collected daily oropharyngeal swabs for quantitative reverse transcriptase-polymerase chain reaction tests using commercial kits. We depicted the cycle threshold (Ct) values for amplification of ORF1ab and N genes from individual patients in age-specific groups in a time series. Results: A total of 480 inpatients were included in the study, with a median age of 59 years (interquartile range, 42 to 78; range, 16 to 106). In the <45-year-old age group, the Ct values for ORF1ab and N gene amplification remained below 35 for 9.0 and 11.5 days, respectively. In the ≥80-year-old age group, the Ct values for ORF1ab and N genes stayed below 35 for 11.5 and 15.0 days, respectively, which was the longest among all age groups. The Ct values for N gene amplification took longer to rise above 35 than those for ORF1ab gene amplification. Conclusion: The time to test negative varied among different age groups, with viral nucleic acid shedding taking longer in older age groups compared to younger age groups. As a result, the time to resolution of Omicron infection increased with increasing age.

Phylogenomic tracing of asymptomatic transmission in a COVID-19 outbreak.

SARS-CoV-2 has caused over 100 million deaths and continues to spread rapidly around the world. Asymptomatic transmission of SARS-CoV-2 is the Achilles' heel of COVID-19 public health control measures. Phylogenomic data on SARS-CoV-2 could provide more direct information about asymptomatic transmission. In this study, using a novel MINERVA sequencing technology, we traced asymptomatic transmission of COVID-19 patients in Beijing, China. One hundred and seventy-eight close contacts were quarantined, and 14 COVID-19 patients were laboratory confirmed by RT-PCR. We provide direct phylogenomic evidence of asymptomatic transmission by constructing the median joining network in the cluster. These data could help us to determine whether the current symptom-based screening should cover asymptomatic persons.

Persistent High Percentage of HLA-DR+CD38high CD8+ T Cells Associated With Immune Disorder and Disease Severity of COVID-19.

Background: The global outbreak of coronavirus disease 2019 (COVID-19) has turned into a worldwide public health crisis and caused more than 100,000,000 severe cases. Progressive lymphopenia, especially in T cells, was a prominent clinical feature of severe COVID-19. Activated HLA-DR+CD38+ CD8+ T cells were enriched over a prolonged period from the lymphopenia patients who died from Ebola and influenza infection and in severe patients infected with SARS-CoV-2. However, the CD38+HLA-DR+ CD8+ T population was reported to play contradictory roles in SARS-CoV-2 infection. Methods: A total of 42 COVID-19 patients, including 32 mild or moderate and 10 severe or critical cases, who received care at Beijing Ditan Hospital were recruited into this retrospective study. Blood samples were first collected within 3 days of the hospital admission and once every 3-7 days during hospitalization. The longitudinal flow cytometric data were examined during hospitalization. Moreover, we evaluated serum levels of 45 cytokines/chemokines/growth factors and 14 soluble checkpoints using Luminex multiplex assay longitudinally. Results: We revealed that the HLA-DR+CD38+ CD8+ T population was heterogeneous, and could be divided into two subsets with distinct characteristics: HLA-DR+CD38dim and HLA-DR+CD38hi. We observed a persistent accumulation of HLA-DR+CD38hi CD8+ T cells in severe COVID-19 patients. These HLA-DR+CD38hi CD8+ T cells were in a state of overactivation and consequent dysregulation manifested by expression of multiple inhibitory and stimulatory checkpoints, higher apoptotic sensitivity, impaired killing potential, and more exhausted transcriptional regulation compared to HLA-DR+CD38dim CD8+ T cells. Moreover, the clinical and laboratory data supported that only HLA-DR+CD38hi CD8+ T cells were associated with systemic inflammation, tissue injury, and immune disorders of severe COVID-19 patients. Conclusions: Our findings indicated that HLA-DR+CD38hi CD8+ T cells were correlated with disease severity of COVID-19 rather than HLA-DR+CD38dim population.

Genomic surveillance of COVID-19 cases in Beijing.

The spread of SARS-CoV-2 in Beijing before May, 2020 resulted from transmission following both domestic and global importation of cases. Here we present genomic surveillance data on 102 imported cases, which account for 17.2% of the total cases in Beijing. Our data suggest that all of the cases in Beijing can be broadly classified into one of three groups: Wuhan exposure, local transmission and overseas imports. We classify all sequenced genomes into seven clusters based on representative high-frequency single nucleotide polymorphisms (SNPs). Genomic comparisons reveal higher genomic diversity in the imported group compared to both the Wuhan exposure and local transmission groups, indicating continuous genomic evolution during global transmission. The imported group show region-specific SNPs, while the intra-host single nucleotide variations present as random features, and show no significant differences among groups. Epidemiological data suggest that detection of cases at immigration with mandatory quarantine may be an effective way to prevent recurring outbreaks triggered by imported cases. Notably, we also identify a set of novel indels. Our data imply that SARS-CoV-2 genomes may have high mutational tolerance.