[Preparation of fluorescent carbon dots by flow-assisted melt polymerization for tetracycline detection in medical wastewater].

Tetracycline (TC) is one of the most important therapeutic drugs that is widely used in hospitals. However, its harmful effects on human health and various ecosystems cannot be ignored. Owing to its poor metabolic activity and low biodegradability, TC commonly discharges as the parent compound and accumulates readily in sludges and soils by precipitation from wastewater, which can induce the evolution of antibiotic-resistant bacteria; therefore, it has been listed as one of the new pollutants with potential ecotoxicological risk. The control measures and environmental management of TC pollutants in environmental water samples require precise determination of TC pollutant concentrations. Carbon dots (CDs) are an emerging type of fluorescent material with numerous advantages such as easy preparation, low cost, low toxicity, and good biocompatibility. Consequently, they have attracted widespread attention in the field of TC detection. Herein, we synthesized TE-CDs with good blue-fluorescence performance via flow-assisted melt polymerization using tricarboxylic acid and ethylenediamine as raw precursors. The morphology and structure of the prepared TE-CDs were characterized. The transmission electron microscopy (TEM) results showed that the prepared TE-CDs were well dispersed, with an average diameter of (2.43±0.48) nm. The X-ray diffraction (XRD) results showed that the TE-CDs had an amorphous carbon structure. Infrared spectroscopy and X-ray photoelectron spectroscopy (XPS) characterizations showed that the surface of the TE-CDs was rich in hydrophilic groups, such as amino, hydroxyl, and carboxyl groups, which indicated that TE-CDs had good water solubility and were advantageous for detecting TC in medical wastewater. Subsequently, the optical properties of the TE-CDs were investigated. The fluorescence emission spectra of the TE-CDs were recorded at various excitation wavelengths. The emission spectra of the TE-CDs exhibited excitation wavelength dependence and when the excitation wavelength changes from 300 nm to 400 nm, their fluorescence intensity decreased to varying degrees. The TE-CDs exhibited optimal fluorescence intensity at an excitation wavelength of 368 nm, while the emission wavelength was 448 nm. TC could effectively quench the blue fluorescence of the CDs, and by utilizing this property, the detection of TC concentration could be achieved. After the addition of TC, the fluorescence of the system immediately reached an extreme value, and no significant change was observed within 10 min. An incubation time of 20 s was selected to obtain precise results. Additionally, the TE-CDs exhibited stable fluorescence intensity over a wide pH range. The fluorescence stability of the TE-CDs was investigated, and no significant change in fluorescence intensity was observed after standing for 10 d, indicating that the prepared TE-CDs had excellent fluorescence stability. The fluorescence intensity of the TE-CDs decreased to varying degrees within the range of 2-200 mg/L TC until complete quenching occurred. TC mass concentration in the range of 4-20 mg/L showed a good linear relationship (R2=0.9978) with the fluorescence quenching intensity of the TE-CDs. The limit of detection was 0.2 mg/L. A preliminary investigation was undertaken to explore the quenching mechanism of the TE-CDs fluorescence by TC. Upon addition of TC, a significant reduction in the fluorescence lifetime of the TE-CDs was observed. During the quenching process, no new substances were observed by UV absorption spectroscopy. Additionally, no significant changes in the 1H NMR spectra of the TE-CDs were noted before and after the addition of TC, indicating the absence of an interaction between the TE-CDs and TC. Therefore, the quenching mechanism may involve dynamic quenching. The selectivity and anti-interference ability of the developed method were evaluated; in the presence of interfering substances, TC quenched the fluorescence of the TE-CDs, indicating that the TE-CDs had good selectivity and anti-interference performance towards TC. The method was applied to the quantitative detection of TC in medical wastewater, with recoveries of 96.5%-119.8% and relative standard deviations of 0.8%-2.6%. In conclusion, the analytical performance of the proposed method is comparable with that of previously reported detection methods; moreover, the method has the advantages of low operational cost, simple preparation process, time-saving, and good repeatability. Therefore, the TE-CDs can be used as chemical sensors for the detection of TC in medical wastewater and have good practical applications.

Morphological, anatomical, and transcriptomics analysis reveals the regulatory mechanisms of cassava plant height development.

BACKGROUND: Cassava is one of three major potato crops and the sixth most important food crop globally. Improving yield remains a primary aim in cassava breeding. Notably, plant height significantly impacts the yield and quality of crops; however, the mechanisms underlying cassava plant height development are yet to be elucidated. RESULTS: In this study, we investigated the mechanisms responsible for cassava plant height development using phenotypic, anatomical, and transcriptomic analyses. Phenotypic and anatomical analysis revealed that compared to the high-stem cassava cultivar, the dwarf-stem cassava cultivar exhibited a significant reduction in plant height and a notable increase in internode tissue xylem area. Meanwhile, physiological analysis demonstrated that the lignin content of dwarf cassava was significantly higher than that of high cassava. Notably, transcriptome analysis of internode tissues identified several differentially expressed genes involved in cell wall synthesis and expansion, plant hormone signal transduction, phenylpropanoid biosynthesis, and flavonoid biosynthesis between the two cassava cultivars. CONCLUSIONS: Our findings suggest that internode tissue cell division, secondary wall lignification, and hormone-related gene expression play important roles in cassava plant height development. Ultimately, this study provides new insights into the mechanisms of plant height morphogenesis in cassava and identifies candidate regulatory genes associated with plant height that can serve as valuable genetic resources for future crop dwarfing breeding.

First report of shoot and leaf blight caused by Botryosphaeria dothidea on Taxus × media in Sichuan province, China.

The genus Taxus is the natural material of the anticancer drug paclitaxel (Xiong et al. 2021). Harvesting sources of paclitaxel from the wild has greatly decreased the population of these trees. One of the taxus species, Taxus × media Rehder, a natural hybrid of taxus trees, has a higher paclitaxel content (Zhou et al. 2019). It has been introduced and cultivated in Sichuan, Chongqing, Yunnan, Zhejiang, Jiangxi, and other places in China. In 2021, approximately 20% of T. media (an average 30% of the affected area per tree) showed obvious shoot and leaf blight symptoms in a plantation of taxus trees (about 40 ha of the planting area), located in Sandaoyan county, Sichuan province, China (GPS, 103°94'60″N, 30°84'97″E). Initially, brown necrotic spots appeared on shoots. Gradually, the spots increased in number, expanded to the leaf attached to the branch, and caused wilting of the shoots and leaves. To identify the pathogen, symptomatic samples were randomly collected. Lesion margins of the diseased leaves and barks were surface sterilized for 1 min in 75% ethanol, rinsed with sterile distilled water three times, dried with sterile filter paper, placed on potato dextrose agar (PDA) amended with streptomycin sulfate (50 mg/liter), and incubated at 28°C in the dark. Six purified fungal isolates were obtained. Collected isolates with similar morphology were described as Botryosphaeria spp. (Zhang et al. 2021). The colonies were initially white, gradually became dark gray with dense erial mycelium after 5 days, and formed black pycnidia (Dimensions, 121.3 to 134.6 µm, n = 5) after 16 days. Conidia were fusiform, aseptate, transparent, and thin-walled (23.6 ± 1.2 × 7.27 ± 1.3 µm, n = 50), similar to B. dothidea (Hattori et al. 2021). For pathogenicity testing, ten 2-year-old seedlings of T. media were selected. Fungal cakes of the isolate Tmsdy-2 were applied to the punctured stems of seedlings and covered with Parafilm. Pieces of sterile medium were used as controls. All the seedlings were incubated at 25 ± 2°C, 50% relative humidity, and 16 h of light in a greenhouse. Four days later, the inoculated seedlings developed brown spots and were blighted in 14 days, with symptoms similar to the original diseased plants. The controls remained healthy. The same fungus was reisolated from the infected tissues and subsequently identified by morphological characteristics and DNA sequence analysis. The pathogenicity test was repeated three times with similar results, confirming Koch's postulates. For molecular identification, the DNA of the isolates was extracted using a Quick-DNA Extraction Kit (Tiangen Biotech, Beijing). The ITS, LSU, SSU, TUB2, and TEF 1-α genes were amplified with the primer pairs ITS1/ITS4, LR0R/LR05, NS1/NS4 (Li et al. 2018), Bt2a/Bt2b, and EF1-728F/EF1-986R (Hattori et al. 2021), respectively. The generated sequences were deposited in GenBank with accession numbers OQ179939 (ITS), OQ179940 (LSU), OQ179942 (SSU), OQ268596 (TUB2), and OQ268597 (TEF 1-α). BLAST analyses showed >99.65% identity with previously deposited sequences of B. dothidea in GenBank. Based on the maximum likelihood method, phylogenetic analysis revealed 100% bootstrap support values with B. dothidea. The fungus was identified as B. dothidea based on morphological and multilocus phylogenetic analyses. To our knowledge, this is the first report of B. dothidea causing shoot and leaf blight of T. media in China. These results will contribute to developing control strategies for this disease.

[Water Quality Change Trend and Risk Analysis of Wuhan Hanjiang River Water Source].

Hanjiang River is closely related to the middle route of the South-to-North Water Diversion Project, the Water Diversion Project from the Hanjiang River to the Wei River, and the Water Diversion Project in Northern Hubei. The Wuhan Hanjiang River water source is one of the important drinking water sources in China; its water quality safety is significant to living and production for millions of residents in Wuhan. Based on data from 2004 to 2021, the water quality variation trend and risk of Wuhan Hanjiang River water source were studied. The results showed that a certain gap existed between the concentrations of some pollutants such as total phosphorus, permanganate index, ammonia nitrogen, and correspondent water quality target, especially for the total phosphorus. The growth of algae in the water source was marginally limited by the concentrations of nitrogen, phosphorus, and silicon. When other factors remained unchanged, diatoms tended to grow rapidly when the water temperature was appropriate (6-12℃). The quality of water upstream had a great impact on the water quality of the Hanjiang water source. There may have been pollutants entering into the reach during the West Lake Water Plant and Zongguan Water Plant. There were differences in the temporal and spatial variation trend of concentrations between permanganate index, total nitrogen, total phosphorus, and ammonia nitrogen. Significant changes in the ratio of nitrogen and phosphorus in the water body will affect the population structure and quantity of planktonic algae and ultimately affect the safety of water quality. The water body in the water source area was generally in the state of medium nutrition to mild eutrophication, and middle eutrophication may have occurred in a few periods. In recent years, the nutritional level of the water source has been on the decline. It is necessary to make an in-depth investigation on the source, quantity, and change trend of pollutants in water sources in order to eliminate potential risks.

Ferroptosis inhibitor liproxstatin-1 alleviates metabolic dysfunction-associated fatty liver disease in mice: potential involvement of PANoptosis.

Ferroptosis is a new form of regulated cell death characterized by excessive iron accumulation and uncontrollable lipid peroxidation. The role of ferroptosis in metabolic dysfunction-associated fatty liver disease (MAFLD) is not fully elucidated. In this study we compared the therapeutic effects of ferroptosis inhibitor liproxstatin-1 (LPT1) and iron chelator deferiprone (DFP) in MAFLD mouse models. This model was established in mice by feeding a high-fat diet with 30% fructose in water (HFHF) for 16 weeks. The mice then received LPT1 (10 mg·kg-1·d-1, ip) or DFP (100 mg·kg-1·d-1, ig) for another 2 weeks. We showed that both LPT1 and DFP treatment blocked the ferroptosis markers ACSL4 and ALOX15 in MAFLD mice. Furthermore, LPT1 treatment significantly reduced the liver levels of triglycerides and cholesterol, lipid peroxidation markers 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA), and ameliorated the expression of lipid synthesis/oxidation genes (Pparα, Scd1, Fasn, Hmgcr and Cpt1a), insulin resistance, mitochondrial ROS content and liver fibrosis. Importantly, LPT1 treatment potently inhibited hepatic apoptosis (Bax/Bcl-xL ratio and TUNEL+ cell number), pyroptosis (cleavages of Caspase-1 and GSDMD) and necroptosis (phosphorylation of MLKL). Moreover, LPT1 treatment markedly inhibited cleavages of PANoptosis-related caspase-8 and caspase-6 in MAFLD mouse liver. In an in vitro MAFLD model, treatment with LPT1 (100 nM) prevented cultured hepatocyte against cell death induced by pro-PANoptosis molecules (TNF-α, LPS and nigericin) upon lipid stress. On the contrary, DFP treatment only mildly attenuated hepatic inflammation but failed to alleviate lipid deposition, insulin resistance, apoptosis, pyroptosis and necroptosis in MAFLD mice. We conclude that ferroptosis inhibitor LPT1 protects against steatosis and steatohepatitis in MAFLD mice, which may involve regulation of PANoptosis, a coordinated cell death pathway that involves apoptosis, pyroptosis and necroptosis. These results suggest a potential link between ferroptosis and PANoptosis.

Lifestyle and Social Factors Exacerbated on the Prevalence of Mood Disorders and Functional Dyspepsia Among Neonatal Nurses in China.

Background: Nursing is a high-stress occupation that can have an impact on mental health, particularly for neonatal nurses. Job-related stress factors and work-related behaviors have played a critical role in nurses' mental health. This study aimed to explore the prevalence of mood disorders and the impact of social factors, lifestyle on mood disorders among neonatal nurses. Methods: A total of 260 participants comprising neonatal nurses and nurses who work in neonatal intensive care units (NICU) were recruited. Data were collected using a validated generalized anxiety disorder questionnaire, patient health questionnaire-9, Pittsburgh sleep quality index, and social factors and lifestyle assessments. Results: In total, 49.23% of neonatal nurses exhibited mood disorders, particularly a combination of depression and anxiety. Female, poor interpersonal relationships and unhappy marital status, preference for smoking, alcohol, irregular diet, and poor sleep were common in neonatology nurses who exhibited mood disorders; preference for coffee and tea were lower in neonatology nurses without mood disorders (all P < 0.05). Interpersonal relationships, marital status, irregular diet, and poor sleep were independent factors associated with mood disorders among neonatal nurses (all P < 0.05). Mood disorders presented as functional dyspepsia (FD) among 50.78% of the participants (P < 0.05). Poor sleep and preference for smoking were common among neonatal nurses who had FD with mood disorders (all P < 0.05). Furthermore, the preference for sugary beverages was lower in participants with FD and mood disorders (P < 0.05). Poor sleep was independently associated with FD with mood disorders in neonatology nurses (P < 0.05). Conclusion: Prevalence of anxiety and depression was higher among neonatal nurses. Furthermore, most cases of mood disorders presented as FD. Thus, social factors and lifestyle have an impact on mood disorders which can manifest through somatic symptoms.

NAD+-boosting therapy alleviates nonalcoholic fatty liver disease via stimulating a novel exerkine Fndc5/irisin.

Rationale: Nicotinamide adenine dinucleotide+ (NAD+)-boosting therapy has emerged as a promising strategy to treat various health disorders, while the underlying molecular mechanisms are not fully understood. Here, we investigated the involvement of fibronectin type III domain containing 5 (Fndc5) or irisin, which is a novel exercise-linked hormone, in the development and progression of nonalcoholic fatty liver disease (NAFLD). Methods: NAD+-boosting therapy was achieved by administrating of nicotinamide riboside (NR) in human and mice. The Fndc5/irisin levels in tissues and blood were measured in NR-treated mice or human volunteers. The therapeutic action of NR against NAFLD pathologies induced by high-fat diet (HFD) or methionine/choline-deficient diet (MCD) were compared between wild-type (WT) and Fndc5-/- mice. Recombinant Fndc5/irisin was infused to NALFD mice via osmotic minipump to test the therapeutic action of Fndc5/irisin. Various biomedical experiments were conducted in vivo and in vitro to know the molecular mechanisms underlying the stimulation of Fndc5/irisin by NR treatment. Results: NR treatment elevated plasma level of Fndc5/irisin in mice and human volunteers. NR treatment also increased Fndc5 expression in skeletal muscle, adipose and liver tissues in mice. In HFD-induced NAFLD mice model, NR displayed remarkable therapeutic effects on body weight gain, hepatic steatosis, steatohepatitis, insulin resistance, mitochondrial dysfunction, apoptosis and fibrosis; however, these actions of NR were compromised in Fndc5-/- mice. Chronic infusion of recombinant Fndc5/irisin alleviated the NAFLD pathological phenotypes in MCD-induced NAFLD mice model. Mechanistically, NR reduced the lipid stress-triggered ubiquitination of Fndc5, which increased Fndc5 protein stability and thus enhanced Fndc5 protein level. Using shRNA-mediated knockdown screening, we found that NAD+-dependent deacetylase SIRT2, rather than other sirtuins, interacts with Fndc5 to decrease Fndc5 acetylation, which reduces Fndc5 ubiquitination and stabilize it. Treatment of AGK2, a selective inhibitor of SIRT2, blocked the therapeutic action of NR against NAFLD pathologies and NR-induced Fndc5 deubiquitination/deacetylation. At last, we identified that the lysine sites K127/131 and K185/187/189 of Fndc5 may contribute to the SIRT2-dependent deacetylation and deubiquitination of Fndc5. Conclusions: The findings from this research for the first time demonstrate that NAD+-boosting therapy reverses NAFLD by regulating SIRT2-deppendent Fndc5 deacetylation and deubiquitination, which results in a stimulation of Fndc5/irisin, a novel exerkine. These results suggest that Fndc5/irisin may be a potential nexus between physical exercise and NAD+-boosting therapy in metabolic pathophysiology.

Lopinavir-ritonavir alone or combined with arbidol in the treatment of 73 hospitalized patients with COVID-19: A pilot retrospective study.

OBJECTIVES: This study aimed to evaluate the antiviral efficacy of lopinavir-ritonavir alone or combined with arbidol in the treatment of hospitalized patients with common coronavirus disease-19 (COVID-19). MATERIALS AND METHODS: In this retrospective observational study, hospitalized COVID-19 patients were identified and divided into two groups based on the antiviral agents during their hospitalization. Patients in group LR were treated with lopinavir-ritonavir 400 mg/100 mg, twice a day, while patients in group LR+Ar were treated with lopinavir-ritonavir 400 mg/100 mg twice a day and arbidol 200 mg three times a day for at least 3 days. Data from these patients were collected from electronic medical record management system. RESULTS: 73 patients were divided into two groups: group LR (34 cases) and group LR+Ar (39 cases), according to the antiviral agents. The overall cure rate of COVID-19 in group LR+Ar and group LR were 92.3% and 97.1%, respectively, with no significant difference (p = 0.62). In a modified intention-to-treat analysis, lopinavir-ritonavir combined with arbidol led to a median time of hospital stay that was shorter by 1.5 days than in group LR (12.5 days vs. 14 days). The percentages of -COVID-19 RNA clearance was 92.3 in group LR and 97.1 in group LR+Ar which was similar to the cure rate. The median time to nucleic acid turning negative = (date of first negative PCR test) - (date of last positive PCR test) was 8.0 days in both groups with no significant difference (p = 0.59). Treatment of lopinavir-ritonavir combined with arbidol did not significantly accelerate main symptom improvement and promote the image absorption of pulmonary inflammation. CONCLUSION: No benefit was observed in the antiviral effect of lopinavir-ritonavir combined with arbidol compared with lopinavir-ritonavir alone in the hospitalized patients with COVID-19. More clinical observations in COVID-19 patients may help to confirm or exclude the effect of antiviral agents.

Excessive ROS production and enhanced autophagy contribute to myocardial injury induced by branched-chain amino acids: Roles for the AMPK-ULK1 signaling pathway and α7nAChR.

BACKGROUNDS AND AIMS: Leucine, isoleucine, and valine are diet derived and essential amino acids that are termed branched-chain amino acids (BCAA). BCAA are widely used as dietary supplements to boost muscle growth and enhance exercise performance. However, the effects of BCAA on myocardial function are largely unknown. This study was designed to investigate whether BCAA affect heart function and, if so, to further explore the underlying molecular basis for the observed effects. METHODS AND RESULTS: C57BL/6J mice were randomly divided into two groups, the control group received solvent (water) and the BCAA group received 2% BCAA dissolved in water, for a successive period of 12 weeks. Compared with control, BCAA treatment significantly increased water consumption without changing body weight or diet consumption; heart tissue BCAA levels were increased, markers representative of myocardial injury in heart tissue including c-reactive protein and cardiac muscle troponin were increased ; and creatine kinase, creatine kinase-MB, and lactate dehydrogenase were increased in serum; severe myocardial fibrosis was observed by Masson staining, which was accompanied by increased reactive oxygen species (ROS) production and decreased superoxide dismutase activity in heart tissue; both p-AMPK and p-ULK1 were significantly increased as was autophagy, judged by the presence of LC3 by western blotting and immunofluorescence, increased numbers of autophagosomes were found by transmission electron microscopy in the BCAA group. In vitro, 20 mmol/L BCAA significantly decreased cell viability and increased the production of ROS, as well as the expression of p-AMPK/AMPK and p-ULK1/ULK1 in cultured H9C2 cells. Treatment with the ROS scavenger N-acetyl-L-cysteine (NAC) improved cell viability and reversed ROS changes. Decreased H9C2 cell viability induced with 20 mmol/L BCAA was reversed by either blocking AMPK or inhibition of ULK1. Furthermore, blocking AMPK significantly decreased p-ULK1/ULK1, while inhibition of ULK1 reversed the enhanced expression of LC3-II/LC3-I induced by BCAA. Excessive ROS production and decreased cell viability induced by BCAA were further confirmed in primary cultured murine cardiomyocytes. Pharmacological activation of α7nAChR with PNU-282987 attenuated BCAA-induced injury in primary murine cardiomyocytes. However, this compound failed to suppress BCAA activation of AMPK and autophagy (LC3-II/I ratio). CONCLUSION: These results provide the first evidence that treatment of mice with BCAA induced myocardial injury by triggering excessive ROS production and by enhancing AMPK-ULK1 pathway-dependent autophagy. These findings suggested that inhibition of either ROS production or autophagy may alleviate myocardial injury induced by BCAA.

The detection and analysis of differential regulatory communities in lung cancer.

The tumorgenesis process of lung cancer involves the regulatory dysfunctions of multiple pathways. Although many signaling pathways have been identified to be associated with lung cancer, there are little quantitative models of how inactions between genes change during the process from normal to cancer. These changes belong to different dynamic co-expressions patterns. We quantitatively analyzed differential co-expression of gene pairs in four datasets. Each dataset included a large number of lung cancer and normal samples. By overlapping their results, we got 14 highly confident gene pairs with consistent co-expression change patterns. Some of they, such as ARHGAP30 and GIMAP4, had been recorded in STRING network database while some of them were novel discoveries, such as C9orf135 and MORN5, TEKT1 and TSPAN1 were positively correlated in both normal and cancer but more correlated in normal than cancer. These gene pairs revealed the underlying mechanisms of lung cancer occurrence.

Design, synthesis and biological evaluation of acridone analogues as novel STING receptor agonists.

STING (Stimulator of Interferon Genes) has become a focal point in immunology research and a target in drug discovery. The discovery of a potent human-STING agonist is expected to revolutionize current anti-virus or cancer immunotherapy. Inspired by the structure and function of murine STING-specific agonists (DMXAA and CMA), we rationally designed and synthesized four series of novel compounds for the enhancement of human sensitivity. In the cell-based assay, we identified six compounds from all the synthetic small molecules: 2g, 9g, and 12b are STING agonists that are efficacious across species, and all have the skeleton of acridone; 1b, 1c, and 12c just function in the murine STING pathway. Notably, 12b exhibits the best activity among the six agonists, and its inductions of both human and murine STING-dependent signalling are similar to that of 2'3'-cGAMP, which is a well-known STING inducer. While a protein assay indicated that 2 g, 9 g, and 12b could activate the pathway by directly binding human STING, 12b also displayed the strongest binding affinity. Additionally, our studies show that 12b can induce faster, more powerful, and more durable responses of assorted cytokines in a native system than 2'3'-cGAMP. Consequently, our team is the first to successfully modify murine STING agonists to obtain human sensitivity, and these results suggest that 12b is a potent direct-human-STING agonist. Additionally, the acridone analogues demonstrate tremendous potential in the treatment of tumours or viral infections.

Oral administration of vancomycin to neonatal mice could alter their immunity and allergic sensibility late in adulthood.

The prevalence of allergy has increased over the past decades, and this may be attributed in part to the intestinal microbiota dysfunction caused by antibiotics during early life. In this study, we evaluated how vancomycin could impair the intestinal microbiota during early life and then, consequently, alter susceptibilities to allergic diseases and related immunity in late adulthood. BALB/c (n=54) neonatal mice were used in this study. Mice in the vancomycin group were orally administered vancomycin for 21 days, while those in the allergy and control groups were perfused with the same volume of saline solution. Then, mice in the allergy and vancomycin groups were immunized with intraperitoneal ovalbumin with alum. At postnatal day 21, vancomycin significantly alter the fecal microbiota, with lower Bacteroidetes and Firmicutes counts and higher Proteobacteria counts. At postnatal day 56, the Bacteroidetes count was still significantly lower in vancomycin-treated mice. The serum IgE level in the control group was significantly lower than that in the vancomycin and allergy groups. The serum interleukin (IL)-6 level and the IL-4/interferon (IFN)-γ values were significantly higher in the vancomycin-treated mice, but the serum IL-17A level was lower than that in the control group. These results indicate that modifications of the intestinal microbiota composition during early life may be, at least in part, the key mechanism underlying the effect of vancomycin that influences the immune function of host animals in the adult stages.

Fusarium graminearum FgCWM1 Encodes a Cell Wall Mannoprotein Conferring Sensitivity to Salicylic Acid and Virulence to Wheat.

Fusarium graminearum causes Fusarium head blight (FHB), a devastating disease of wheat. Salicylic acid (SA) is involved in the resistance of wheat to F. graminearum. Cell wall mannoprotein (CWM) is known to trigger defense responses in plants, but its role in the pathogenicity of F. graminearum remains unclear. Here, we characterized FgCWM1 (FG05_11315), encoding a CWM in F. graminearum. FgCWM1 was highly expressed in wheat spikes by 24 h after initial inoculation and was upregulated by SA. Disruption of FgCWM1 (ΔFgCWM1) reduced mannose and protein accumulation in the fungal cell wall, especially under SA treatment, and resulted in defective fungal cell walls, leading to increased fungal sensitivity to SA. The positive role of FgCWM1 in mannose and protein accumulation was confirmed by its expression in Saccharomyces cerevisiae. Compared with wild type (WT), ΔFgCWM1 exhibited reduced pathogenicity toward wheat, but it produced the same amount of deoxynivalenol both in culture and in spikes. Complementation of ΔFgCWM1 with FgCWM1 restored the WT phenotype. Localization analyses revealed that FgCWM1 was distributed on the cell wall, consistent with its structural role. Thus, FgCWM1 encodes a CWM protein that plays an important role in the cell wall integrity and pathogenicity of F. graminearum.

Design, Synthesis and Biological Evaluation of 1-Phenyl-2-(phenylamino) Ethanone Derivatives as Novel MCR-1 Inhibitors.

Polymyxins are considered to be the last-line antibiotics that are used to treat infections caused by multidrug-resistant (MDR) gram-negative bacteria; however, the plasmid-mediated transferable colistin resistance gene (mcr-1) has rendered polymyxins ineffective. Therefore, the protein encoded by mcr-1, MCR-1, could be a target for structure-based design of inhibitors to tackle polymyxins resistance. Here, we identified racemic compound 3 as a potential MCR-1 inhibitor by virtual screening, and 26 compound 3 derivatives were synthesized and evaluated in vitro. In the cell-based assay, compound 6g, 6h, 6i, 6n, 6p, 6q, and 6r displayed more potent activity than compound 3. Notably, 25 µΜ of compound 6p or 6q combined with 2 µg·mL-1 colistin could completely inhibit the growth of BL21(DE3) expressing mcr-1, which exhibited the most potent activity. In the enzymatic assay, we elucidate that 6p and 6q could target the MCR-1 to inhibit the activity of the protein. Additionally, a molecular docking study showed that 6p and 6q could interact with Glu246 and Thr285 via hydrogen bonds and occupy well the cavity of the MCR-1 protein. These results may provide a potential avenue to overcome colistin resistance, and provide some valuable information for further investigation on MCR-1 inhibitors.

Re-acquisition of the brittle rachis trait via a transposon insertion in domestication gene Q during wheat de-domestication.

De-domestication is a unique evolutionary process during which crops re-acquire wild-like traits to survive and persist in agricultural fields without the need for human cultivation. The re-acquisition of seed dispersal mechanisms is crucial for crop de-domestication. Common wheat is an important cereal crop worldwide. Tibetan semi-wild wheat is a potential de-domesticated common wheat subspecies. However, the crucial genes responsible for its brittle rachis trait have not been identified. Genetic mapping, functional analyses and phylogenetic analyses were completed to identify the gene associated with Qbr.sau-5A, which is a major locus for the brittle rachis trait of Tibetan semi-wild wheat. The cloned Qbr.sau-5A gene is a new Q allele (Qt ) with a 161-bp transposon insertion in exon 5. Although Qt is expressed normally, its encoded peptide lacks some key features of the APETALA2 family. The abnormal functions of Qt in developing wheat spikes result in brittle rachises. Phylogenetic and genotyping analyses confirmed that Qt originated from Q in common wheat and is naturally distributed only in Tibetan semi-wild wheat populations. The identification of Qt provides new evidence regarding the origin of Tibetan semi-wild wheat, and new insights into the re-acquisition of wild traits during crop de-domestication.

Transcriptional reference map of hormone responses in wheat spikes.

BACKGROUND: Phytohormones are key regulators of plant growth, development, and signalling networks involved in responses to diverse biotic and abiotic stresses. Transcriptional reference maps of hormone responses have been reported for several model plant species such as Arabidopsis thaliana, Oryza sativa, and Brachypodium distachyon. However, because of species differences and the complexity of the wheat genome, these transcriptome data are not appropriate reference material for wheat studies. RESULTS: We comprehensively analysed the transcriptomic responses in wheat spikes to seven phytohormones, including indole acetic acid (IAA), gibberellic acid (GA), abscisic acid (ABA), ethylene (ET), cytokinin (CK), salicylic acid (SA), and methyl jasmonic acid (MeJA). A total of 3386 genes were differentially expressed at 24 h after the hormone treatments. Furthermore, 22.7% of these genes exhibited overlapping transcriptional responses for at least two hormones, implying there is crosstalk among phytohormones. We subsequently identified genes with expression levels that were significantly and differentially induced by a specific phytohormone (i.e., hormone-specific responses). The data for these hormone-responsive genes were then compared with the transcriptome data for wheat spikes exposed to biotic (Fusarium head blight) and abiotic (water deficit) stresses. CONCLUSION: Our data were used to develop a transcriptional reference map of hormone responses in wheat spikes.

Functional Analysis of FgNahG Clarifies the Contribution of Salicylic Acid to Wheat (Triticum aestivum) Resistance against Fusarium Head Blight.

Salicylic acid (SA) is a key defense hormone associated with wheat resistance against Fusarium head blight, which is a severe disease mainly caused by Fusarium graminearum. Although F. graminearum can metabolize SA, it remains unclear how this metabolic activity affects the wheat⁻F. graminearum interaction. In this study, we identified a salicylate hydroxylase gene (FG05_08116; FgNahG) in F. graminearum. This gene encodes a protein that catalyzes the conversion of SA to catechol. Additionally, FgNahG was widely distributed within hyphae. Disrupting the FgNahG gene (ΔFgNahG) led to enhanced sensitivity to SA, increased accumulation of SA in wheat spikes during the early infection stage and inhibited development of head blight symptoms. However, FgNahG did not affect mycotoxin production. Re-introducing a functional FgNahG gene into the ΔFgNahG mutant recovered the wild-type phenotype. Moreover, the expression of FgNahG in transgenic Arabidopsis thaliana decreased the SA concentration and the resistance of leaves to F. graminearum. These results indicate that the endogenous SA in wheat influences the resistance against F. graminearum. Furthermore, the capacity to metabolize SA is an important factor affecting the ability of F. graminearum to infect wheat plants.

Mechanisms of wheat (Triticum aestivum) grain storage proteins in response to nitrogen application and its impacts on processing quality.

Basis for the effects of nitrogen (N) on wheat grain storage proteins (GSPs) and on the establishment of processing quality are far from clear. The response of GSPs and processing quality parameters to four N levels of four common wheat cultivars were investigated at two sites over two growing seasons. Except gluten index (GI), processing quality parameters as well as GSPs quantities were remarkably improved by increasing N level. N level explained 4.2~59.2% and 10.4~80.0% variability in GSPs fractions and processing quality parameters, respectively. The amount of N remobilized from vegetative organs except spike was significantly increased when enhancing N application. GSPs fractions and processing quality parameters except GI were only highly and positively correlated with the amount of N remobilized from stem with sheath. N reassimilation in grain was remarkably strengthened by the elevated activity and expression level of glutamine synthetase. Transcriptome analysis showed the molecular mechanism of seeds in response to N levels during 10~35 days post anthesis. Collectively, we provided comprehensive understanding of N-responding mechanisms with respect to wheat processing quality from N source to GSPs biosynthesis at the agronomic, physiological and molecular levels, and screened candidate genes for quality breeding.