CaSTH2 disables CaWRKY40 from activating pepper thermotolerance and immunity against Ralstonia solanacearum via physical interaction.

CaWRKY40 coordinately activates pepper immunity against Ralstonia solanacearum infection (RSI) and high temperature stress (HTS), forms positive feedback loops with other positive regulators and is promoted by CaWRKY27b/CaWRKY28 through physical interactions; however, whether and how it is regulated by negative regulators to function appropriately remain unclear. Herein, we provide evidence that CaWRKY40 is repressed by a SALT TOLERANCE HOMOLOG2 in pepper (CaSTH2). Our data from gene silencing and transient overexpression in pepper and epoptic overexpression in Nicotiana benthamiana plants showed that CaSTH2 acted as negative regulator in immunity against RSI and thermotolerance. Our data from BiFC, CoIP, pull down, and MST indicate that CaSTH2 interacted with CaWRKY40, by which CaWRKY40 was prevented from activating immunity or thermotolerance-related genes. It was also found that CaSTH2 repressed CaWRKY40 at least partially through blocking interaction of CaWRKY40 with CaWRKY27b/CaWRKY28, but not through directly repressing binding of CaWRKY40 to its target genes. The results of study provide new insight into the mechanisms underlying the coordination of pepper immunity and thermotolerance.

Recovery of organic matters by activated sludge from municipal wastewater: Performance and characterization.

Municipal wastewater treatment processes consume a significant amount of energy and generate substantial carbon emissions. However, organic matters existing in municipal wastewater hold the potential as a valuable carbon source. Activated sludge has the potential to capture and recover the organic matters, thereby enriching carbon sources and facilitating subsequent sludge anaerobic digestion as well as in line with the concept of sustainable development. Based on above, this study investigated the enrichment and recovery characteristics and mechanisms of activated sludge adsorption on carbon sources in municipal wastewater, while optimizing the recovery conditions. The results indicated that insoluble organic matters, as well as a fraction of dissolved organic matters, can be effective recovered within approximately 40 min. Specifically, 74.1% of insoluble organic matters and 25.8% of soluble organic matters were successfully captured by the activated sludge, resulting in a 5.0% increase in sludge organic matter content. Moreover, activated sludge demonstrated remarkable recovery of particulate organic matters across various particle sizes, particularly larger particles (>5 µm) with high protein content. Notably, the dissolved biodegradable organics such as tryptophan and tyrosine protein-like substances according to 3D-EEM and lipids, proteins/amino sugars, and carbohydrates according to FT-ICR MS can be effectively recovered. Finally, the study revealed that the recovery of organic matters from the wastewater by activated sludge followed the pseudo-second-order kinetics model, with surface binding, hydrogen bonding and interparticle diffusion in sludge flocs as the primary adsorption mechanisms. This approach had abroad application prospects for improving the profitability of wastewater treatment plants.

Differential CaKAN3-CaHSF8 associations underlie distinct immune and heat responses under high temperature and high humidity conditions.

High temperature and high humidity (HTHH) conditions increase plant susceptibility to a variety of diseases, including bacterial wilt in solanaceous plants. Some solanaceous plant cultivars have evolved mechanisms to activate HTHH-specific immunity to cope with bacterial wilt disease. However, the underlying mechanisms remain poorly understood. Here we find that CaKAN3 and CaHSF8 upregulate and physically interact with each other in nuclei under HTHH conditions without inoculation or early after inoculation with R. solanacearum in pepper. Consequently, CaKAN3 and CaHSF8 synergistically confer immunity against R. solanacearum via activating a subset of NLRs which initiates immune signaling upon perception of unidentified pathogen effectors. Intriguingly, when HTHH conditions are prolonged without pathogen attack or the temperature goes higher, CaHSF8 no longer interacts with CaKAN3. Instead, it directly upregulates a subset of HSP genes thus activating thermotolerance. Our findings highlight mechanisms controlling context-specific activation of high-temperature-specific pepper immunity and thermotolerance mediated by differential CaKAN3-CaHSF8 associations.

PMT6 Is Required for SWC4 in Positively Modulating Pepper Thermotolerance.

High temperature stress (HTS), with growth and development impairment, is one of the most important abiotic stresses frequently encountered by plants, in particular solanacaes such as pepper, that mainly distribute in tropical and subtropical regions. Plants activate thermotolerance to cope with this stress; however, the underlying mechanism is currently not fully understood. SWC4, a shared component of SWR1- and NuA4 complexes implicated in chromatin remodeling, was previously found to be involved in the regulation of pepper thermotolerance, but the underlying mechanism remains poorly understood. Herein, PMT6, a putative methyltranferase was originally found to interact with SWC4 by co-immunoprecipitation (Co-IP)-combined LC/MS assay. This interaction was further confirmed by bimolecular fluorescent complimentary (BiFC) and Co-IP assay, and PMT6 was further found to confer SWC4 methylation. By virus-induced gene silencing, it was found that PMT6 silencing significantly reduced pepper basal thermotolerance and transcription of CaHSP24 and significantly reduced the enrichment of chromatin-activation-related H3K9ac, H4K5ac, and H3K4me3 in TSS of CaHSP24, which was previously found to be positively regulated by CaSWC4. By contrast, the overexpression of PMT6 significantly enhanced basal thermotolerance of pepper plants. All these data indicate that PMT6 acts as a positive regulator in pepper thermotolerance, likely by methylating SWC4.

Evaluation of the potential anti-soybean allergic activity of different forms of Lactobacillus delbrueckii subsp. bulgaricus based on cell model in vitro.

Live, inactivated Lactobacillus or their metabolites have various beneficial functions, which may alleviate food allergy. This study aimed to investigate the intervention effects of three forms of Lactobacillus delbrueckii subsp. bulgaricus (Ld) on cell degranulation, intestinal barrier function, and intestinal mucosal immunity against soybean allergy. First, the intervention effect of Ld on cell degranulation was investigated using the KU812 cell degranulation model. Then, the Caco-2 cell inflammation model was used to evaluate their anti-inflammatory capacity, and the cell monolayer model was constructed to test the protective effects of different forms of Ld on the intestinal barrier. Finally, mesenteric lymph node (MLN) cells from mice were used to assess the ability of different forms of Ld to regulate the balance of cytokines associated with food allergy in the immune tissue of the intestinal mucosa. Results showed that live bacteria and heat-inactivated bacteria could inhibit the degranulation of KU812 cells, mainly by significantly inhibiting the release of histamine, IL-6 and TNF-α. Both live bacteria and heat-inactivated bacteria could also suppress the increase of IL-6 and IL-8 in Caco-2 cells induced by lipopolysaccharide (LPS). The culture supernatant of bacteria and live bacteria showed better ability to maintain the integrity and permeability of the intestinal epithelial barrier. In addition, heat-inactivated bacteria could return the values of IFN-γ and IL-10 to normal levels and restore the balance of IFN-γ/IL-4, thereby reversing the immune deviation of MLN cells. Therefore, three forms of Ld have potential for the treatment of soybean allergy.

Effects of Different Doses of Rimazolam on Oxidative Stress Level Sedation Score and Recovery Time of Patients during Hip Replacement.

The effects of Rimazolam on oxidative stress level, sedation score, and recovery time of patients under different doses in hip replacement are investigated. A total of 100 patients who underwent hip arthroplasty in our hospital from September 2020 to May 2022 are selected as the study subjects. According to the random number table method, they are divided into observation group and control group. The observation group is given 0.3 mg/kg Rimazolam, and the control group is given 0.4 mg/kg Rimazolam. Oxidative stress level, hemodynamic indexes at different time after anesthesia, sedation score and sedation depth, Visual Analogue Scale/Score (VAS) score and incidence of adverse reactions are observed in 2 groups. The correlation of sedation score with oxidative stress index and pain degree is analyzed. The experimental results show that Rimazolam has a good anesthesia induction effect in hip replacement, but the low dose (0.3 mg/kg) can reduce the level of oxidative stress in patients, has a better sedation effect and can maintain the stability of patients' hemodynamic indicators, and has a positive effect on relieving postoperative pain in patients. [Clinical Trial Registration Number- ChiCTR2000038548].

Effects of Different Doses of Dex Anesthesia on Inflammatory Factors and Hemodynamics in Patients Undergoing Neurosurgery and Its Relationship with RSS Score.

The effects of different doses of dexmedetomidine (Dex) anesthesia on inflammatory factors and hemodynamics in patients undergoing neurosurgery and its relationship with RSS scores are analyzed. A total of 102 patients who received neurosurgery in our hospital from March 2021 to March 2022 are selected. According to different intraoperative Dex dose, the enrolled patients are randomly divided into low-, medium-, and high-dose groups, 34 patients in each group. In this study, hemodynamic indexes, inflammatory factors, anesthesia effect, and the Ramsay sedation scale (RSS) score are compared in each groups. The experimental results show that in neurosurgery, compared with low-dose Dex, high-dose Dex can better maintain patients' intraoperative hemodynamic parameters and effectively inhibit postoperative inflammatory response, but postoperative awakening time is also relatively prolonged, while medium-dose Dex can effectively control patients' awakening time.

CabZIP23 Integrates in CabZIP63-CaWRKY40 Cascade and Turns CabZIP63 on Mounting Pepper Immunity against Ralstonia solanacearum via Physical Interaction.

CabZIP63 and CaWRKY40 were previously found to be shared in the pepper defense response to high temperature stress (HTS) and to Ralstonia solanacearum inoculation (RSI), forming a transcriptional cascade. However, how they activate the two distinct defense responses is not fully understood. Herein, using a revised genetic approach, we functionally characterized CabZIP23 in the CabZIP63-CaWRKY40 cascade and its context specific pepper immunity activation against RSI by interaction with CabZIP63. CabZIP23 was originally found by immunoprecipitation-mass spectrometry to be an interacting protein of CabZIP63-GFP; it was upregulated by RSI and acted positively in pepper immunity against RSI by virus induced gene silencing in pepper plants, and transient overexpression in Nicotiana benthamiana plants. By chromatin immunoprecipitation (ChIP)-qPCR and electrophoresis mobility shift assay (EMSA), CabZIP23 was found to be directly regulated by CaWRKY40, and CabZIP63 was directly regulated by CabZIP23, forming a positive feedback loop. CabZIP23-CabZIP63 interaction was confirmed by co-immunoprecipitation (CoIP) and bimolecular fluorescent complimentary (BiFC) assays, which promoted CabZIP63 binding immunity related target genes, including CaPR1, CaNPR1 and CaWRKY40, thereby enhancing pepper immunity against RSI, but not affecting the expression of thermotolerance related CaHSP24. All these data appear to show that CabZIP23 integrates in the CabZIP63-CaWRKY40 cascade and the context specifically turns it on mounting pepper immunity against RSI.

Characterization of the protein structure of soymilk fermented by Lactobacillus and evaluation of its potential allergenicity based on the sensitized-cell model.

This study aimed to investigate the effects of fermented soymilk (FSM) with Lactobacillus brevis CICC 23,474 and L. brevis CICC 23,470 on the structural changes and allergenicity of major allergenic proteins in soymilk (SM). Spectroscopy and liquid chromatograph-tandem mass spectrometer (LC-MS/MS) were used to characterize changes in protein spatial structure and epitopes. The antigenicity and potential allergenicity were evaluated by immunoblotting, enzyme-linked immunosorbent assay (ELISA) and KU812 cell degranulation assay. Results suggested that the advanced structure of proteins was destroyed. Antigenicity was also significantly reduced, and five human IgE-binding linear epitopes (i.e., E5-E33, R27-S41, D414-A437, G253-I265 and V449-S471) were destroyed by fermentation. Furthermore, after in vitro simulated gastrointestinal digestion, FSM showed lower IgG/IgE-binding capacity and weaker degranulation ability of KU812 cells. All these findings demonstrated that fermentation with Lactobacillus can destroy the conformational and linear epitopes of proteins and reduce the potential allergenicity of SM.

Solanaceous plants switch to cytokinin-mediated immunity against Ralstonia solanacearum under high temperature and high humidity.

Plant diseases generally tend to be more serious under conditions of high temperature and high humidity (HTHH) than under ambient temperature, but plant immunity against pathogen attacks under HTHH remains elusive. Herein, we used pepper as an example to study how Solanaceae cope with Ralstonia solanacearum infection (RSI) under HTHH by performing RNA-seq combined with the reverse genetic method. The result showed that immunities mediated by salicylic acid (SA) and jasmonic acid (JA) in pepper roots were activated by RSI under ambient temperature. However, upon RSI under HTHH, JA signalling was blocked and SA signalling was activated early but its duration was greatly shortened in pepper roots, instead, expression of CaIPT5 and Glutathione S-transferase encoding genes, as well as endogenous content of trans-Zeatin, were enhanced. In addition, by silencing in pepper plants and overexpression in Nicotiana benthamiana, CaIPT5 was found to act positively in the immune response to RSI under HTHH in a way related to CaPRP1 and CaMgst3. Furthermore, the susceptibility of pepper, tomato and tobacco to RSI under HTHH was significantly reduced by exogenously applied tZ, but not by either SA or MeJA. All these data collectively suggest that pepper employs cytokinin-mediated immunity to cope with RSI under HTHH.

Discovery of new genetic loci for male sexual orientation in Han population.

Epidemiological studies have demonstrated that the genetic factors partly influence the development of same-sex sexual behavior, but most genetic studies have focused on people of primarily European ancestry, potentially missing important biological insights. Here, we performed a two-stage genome-wide association study (GWAS) with a total sample of 1478 homosexual males and 3313 heterosexual males in Han Chinese populations and identified two genetic loci (rs17320865, Xq27.3, FMR1NB, Pmeta = 8.36 × 10-8, OR = 1.29; rs7259428, 19q12, ZNF536, Pmeta = 7.58 × 10-8, OR = 0.75) showing consistent association with male sexual orientation. A fixed-effect meta-analysis including individuals of Han Chinese (n = 4791) and European ancestries (n = 408,995) revealed 3 genome-wide significant loci of same-sex sexual behavior (rs9677294, 2p22.1, SLC8A1, Pmeta = 1.95 × 10-8; rs2414487, 15q21.3, LOC145783, Pmeta = 4.53 × 10-9; rs2106525, 7q31.1, MDFIC, Pmeta = 6.24 × 10-9). These findings may provide new insights into the genetic basis of male sexual orientation from a wider population scope. Furthermore, we defined the average ZNF536-immunoreactivity (ZNF536-ir) concentration in the suprachiasmatic nucleus (SCN) as lower in homosexual individuals than in heterosexual individuals (0.011 ± 0.001 vs 0.021 ± 0.004, P = 0.013) in a postmortem study. In addition, compared with heterosexuals, the percentage of ZNF536 stained area in the SCN was also smaller in the homosexuals (0.075 ± 0.040 vs 0.137 ± 0.103, P = 0.043). More homosexual preference was observed in FMR1NB-knockout mice and we also found significant differences in the expression of serotonin, dopamine, and inflammation pathways that were reported to be related to sexual orientation when comparing CRISPR-mediated FMR1NB knockout mice to matched wild-type target C57 male mice.

CaAIL1 Acts Positively in Pepper Immunity against Ralstonia solanacearum by Repressing Negative Regulators.

APETALA2 (AP2) subfamily transcription factors participate in plant growth and development, but their roles in plant immunity remain unclear. Here, we discovered that the AP2 transcription factor CaAIL1 functions in immunity against Ralstonia solanacearum infection (RSI) in pepper (Capsicum annuum). CaAIL1 expression was upregulated by RSI, and loss- and gain-of-function assays using virus-induced gene silencing and transient overexpression, respectively, revealed that CaAIL1 plays a positive role in immunity to RSI in pepper. Chromatin immunoprecipitation sequencing (ChIP-seq) uncovered a subset of transcription-factor-encoding genes, including CaRAP2-7, CaGATA17, CaGtf3a and CaTCF25, that were directly targeted by CaAIL1 via their cis-elements, such as GT or AGGCA motifs. ChIP-qPCR and electrophoretic mobility shift assays confirmed these findings. These genes, encoding transcription factors with negative roles in immunity, were repressed by CaAIL1 during pepper response to RSI, whereas genes encoding positive immune regulators such as CaEAS were derepressed by CaAIL1. Importantly, we showed that the atypical EAR motif (LXXLXXLXX) in CaAIL1 is indispensable for its function in immunity. These findings indicate that CaAIL1 enhances the immunity of pepper against RSI by repressing a subset of negative immune regulators during the RSI response through its binding to several cis-elements in their promoters.

An outbreak of pulmonary tuberculosis and a follow-up investigation of latent tuberculosis in a high school in an eastern city in China, 2016-2019.

BACKGROUND: In October 2016, a senior high school student was diagnosed with sputum-smear positive [SS(+)] pulmonary tuberculosis (TB). We conducted an investigation of an outbreak in the school, including among students and teachers diagnosed with latent TB, who we followed until July 2019. METHODS: We defined latent TB infection (LTBI) as a tuberculin skin test (TST) induration of 15mm or larger; probable TB as a chest radiograph indicative of TB plus productive cough/hemoptysis for at least 2 weeks, or TST induration of 15mm or larger; and confirmed TB as two or more positive sputum smears or one positive sputum smear plus a chest radiograph indicative of TB or culture positive with M. tuberculosis. We conducted mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing based on 24 loci in the isolates. RESULTS: Between October 2016 and July 2019, we identified 52 cases, including nine probable, six confirmed, and 37 LTBI cases. The index case-student had attended school continuously despite having TB symptoms for almost three months before being diagnosed with TB. We obtained three isolates from classmates of the index case in 2016; all had identical MIRU-VNTR alleles with the index case. The LTBI rate was lower among students (7.41%, 30/405) than among teachers (26.92%, 7/26) (rate ratio [RR] = 0.28, 95% confidential interval [CI]: 0.13-0.57). Among the 17 students who had latent TB and refused prophylaxis in October 2016, 23.53% (4/17) became probable/confirmed cases by July 2019. None of the six teachers who also refused prophylaxis became probable or confirmed cases. Of the 176 students who were TST(-) in October 2016, 1.70% (3/176) became probable/confirmed cases, and among the 20 teachers who were TST(-), 1 became a probable case. CONCLUSIONS: Delayed diagnosis of TB in the index patient may have contributed to the start of this outbreak; lack of post-exposure chemoprophylaxis facilitated spread of the outbreak. Post-exposure prophylaxis is strongly recommended for all TST-positive students; TST-negative students exposed to an SS(+) case should be followed up regularly so that prophylaxis can be started if LTBI is detected.

Effect of transglutaminase cross-linking on the allergenicity of tofu based on a BALB/c mouse model.

Soybean products are limited in terms of safe consumption because of the sensitization of raw materials. In this study, the allergenicity of cross-linked tofu with microbial transglutaminase (MTG) was evaluated on the basis of a BALB/c mouse model. The mice were randomly divided into five groups. Cholera toxin was used as an adjuvant to sensitize the mice through intragastric administration, and tofu was given orally to investigate its sensitization effect on the mice. The allergy symptoms, body temperature, and weight of the mice were detected. The immunoglobulin E (IgE), immunoglobulin G (IgG), and spleen cytokines of the mice were determined through an enzyme-linked immunosorbent assay. The regulation of the differentiation balance of the different subsets of splenic T lymphocyte (Th1, Th2) and regulatory T cells (Tregs) in the mice was measured through flow cytometry. Results showed that the mice administered with MTG-cross-linked tofu had fewer allergic symptoms compared with those of the control group. The concentrations of serum-specific IgE and IgG, plasma histamine, and mast cell protease 1 (mMCP-1) significantly decreased. The Th2-related cytokine levels reduced, and the IFN-γ levels increased. The proportion of Th2 cells decreased, and the proportion of CD4+CD25+Foxp+ Tregs increased as the percentage of Th1 cells increased. Therefore, the sensitization of enzymatic cross-linked tofu decreased.

[Soil organic carbon storage and its spatial distribution in Henan Province].

By using 1: 200 000 soil database, the soil organic carbon storage in Henan Province was estimated, and its spatial distribution was analyzed. The results showed that in this province, soil organic carbon storage was about 10.27 x 10(8) t, accounting for 1.15% of the total in China, and its density was 7.46 kg m(-2) on average, being lower than that (9.60 kg m(-2)) in this country. The top four soil types in organic carbon storage were fluvo-aquic soil, cinnamon soil, skeletal soil, and yellow cinnamon soil, with the storage all being higher than 1.0 x 10(8) t and totally taken up 69.65% of that in Henan. The organic carbon density was the highest in bog soil (24.54 kg m(-2)), followed by in mountain meadow soil (17.69 kg m(-2)) and brown soil (14.64 kg m(-2)). These three soil types mainly distributed in the mountainous areas of west Henan, and the sum of their organic carbon storage was only accounted for 6.34% of the total in the province. The organic carbon density was the lowest in rocky soil (1.32 kg m(-2)) and aeolian sandy soil (1.38 kg m(-2)). In general, the density of soil organic carbon in Henan province mostly varied from 5 kg m(-2) to 10 kg m(-2).