Chemically stable fluorescent proteins for advanced microscopy.

We report the rational engineering of a remarkably stable yellow fluorescent protein (YFP), 'hyperfolder YFP' (hfYFP), that withstands chaotropic conditions that denature most biological structures within seconds, including superfolder green fluorescent protein (GFP). hfYFP contains no cysteines, is chloride insensitive and tolerates aldehyde and osmium tetroxide fixation better than common fluorescent proteins, enabling its use in expansion and electron microscopies. We solved crystal structures of hfYFP (to 1.7-Å resolution), a monomeric variant, monomeric hyperfolder YFP (1.6 Å) and an mGreenLantern mutant (1.2 Å), and then rationally engineered highly stable 405-nm-excitable GFPs, large Stokes shift (LSS) monomeric GFP (LSSmGFP) and LSSA12 from these structures. Lastly, we directly exploited the chemical stability of hfYFP and LSSmGFP by devising a fluorescence-assisted protein purification strategy enabling all steps of denaturing affinity chromatography to be visualized using ultraviolet or blue light. hfYFP and LSSmGFP represent a new generation of robustly stable fluorescent proteins developed for advanced biotechnological applications.

Isolation of Fluorescent 2π-Aromatic 1,3-Disiladiboretenes.

Herein, we reported the isolation of 2π-aromatic disiladiboretenes (L2Si2B2Ph2) [L = ArC(NtBu)2, Ar = Ph (1), Mes (2)], which have been synthesized from the straightforward reduction of silylene-borane adducts (LSiX → BX2Ph) [X = Cl, Br] with potassium graphite (KC8). X-ray diffraction analysis of 1 and 2 revealed that the Si2B2 units are completely planar, and DFT calculations suggested delocalization of 2π-electrons over the Si2B2 rings. Moreover, their photophysical properties and reactivity toward sulfur were also investigated in detail.

Integrated transcriptome and DNA methylome analysis reveal the biological base of increased resistance to gray leaf spot and growth inhibition in interspecific grafted tomato scions.

BACKGROUND: Grafting is widely used as an important agronomic approach to deal with environmental stresses. However, the molecular mechanism of grafted tomato scions in response to biotic stress and growth regulation has yet to be fully understood. RESULTS: This study investigated the resistance and growth performance of tomato scions grafted onto various rootstocks. A scion from a gray leaf spot-susceptible tomato cultivar was grafted onto tomato, eggplant, and pepper rootstocks, creating three grafting combinations: one self-grafting of tomato/tomato (TT), and two interspecific graftings, namely tomato/eggplant (TE) and tomato/pepper (TP). The study utilized transcriptome and DNA methylome analyses to explore the regulatory mechanisms behind the resistance and growth traits in the interspecific graftings. Results indicated that interspecific grafting significantly enhanced resistance to gray leaf spot and improved fruit quality, though fruit yield was decreased compared to self-grafting. Transcriptome analysis demonstrated that, compared to self-grafting, interspecific graftings triggered stronger wounding response and endogenous immune pathways, while restricting genes related to cell cycle pathways, especially in the TP grafting. Methylome data revealed that the TP grafting had more hypermethylated regions at CHG (H = A, C, or T) and CHH sites than the TT grafting. Furthermore, the TP grafting exhibited increased methylation levels in cell cycle related genes, such as DNA primase and ligase, while several genes related to defense kinases showed decreased methylation levels. Notably, several kinase transcripts were also confirmed among the rootstock-specific mobile transcripts. CONCLUSIONS: The study concludes that interspecific grafting alters gene methylation patterns, thereby activating defense responses and inhibiting the cell cycle in tomato scions. This mechanism is crucial in enhancing resistance to gray leaf spot and reducing growth in grafted tomato scions. These findings offer new insights into the genetic and epigenetic contributions to agronomic trait improvements through interspecific grafting.

Transient non-native hydrogen bonds promote activation of a signaling protein.

Phosphorylation is a common mechanism for activating proteins within signaling pathways. Yet, the molecular transitions between the inactive and active conformational states are poorly understood. Here we quantitatively characterize the free-energy landscape of activation of a signaling protein, nitrogen regulatory protein C (NtrC), by connecting functional protein dynamics of phosphorylation-dependent activation to protein folding and show that only a rarely populated, pre-existing active conformation is energetically stabilized by phosphorylation. Using nuclear magnetic resonance (NMR) dynamics, we test an atomic scale pathway for the complex conformational transition, inferred from molecular dynamics simulations (Lei et al., 2009). The data show that the loss of native stabilizing contacts during activation is compensated by non-native transient atomic interactions during the transition. The results unravel atomistic details of native-state protein energy landscapes by expanding the knowledge about ground states to transition landscapes.

The Burden of Occupational Noise-Induced Hearing Loss From 1990 to 2019: An Analysis of Global Burden of Disease Data.

OBJECTIVES: The relationship between long-term exposure to occupational noise and hearing loss has been extensively documented. We aimed to assess spatial and temporal changes in the burden of occupational noise-induced hearing loss (ONIHL) in 204 countries and territories with varying socio-demographic indexes (SDI) from 1990 to 2019. DESIGN: Temporal and spatial trends in age-standardized disability-adjusted life year rates (ASDR) for ONIHL were estimated by sex, age, SDI level, country, and geographic region from 1990 to 2019. We used the Joinpoint model to calculate annual average percentage changes to assess such trends and projected trends in ASDR for ONIHL globally and across different income regions from 2020 to 2044 using an age-period-cohort model. We fitted the relationship between ASDR and SDI, ASDR and healthcare access and quality index, respectively. RESULTS: Overall, the global burden of ONIHL has decreased since 1990, especially in middle and lower SDI regions. In 2019, the global ASDR for ONIHL was 84.23 (95% confidence interval: 57.46 to 120.52) per 100,000 population. From 1990 to 2019, the global ASDR for ONIHL decreased by 1.72% (annual average percentage change = -0.05, 95% confidence interval: -0.07 to -0.03). Our projections showed a decreasing trend in the global ONIHL burden until 2044. ASDR and SDI (R = -0.8, p < 0.05), ASDR and healthcare access and quality index (R = -0.75, p < 0.05) showed significant negative correlations. CONCLUSIONS: The global ONIHL burden has decreased over the past three decades, especially in regions with middle and lower SDI levels. However, the global ONIHL burden still remained severe in 2019, notably among males, the middle-aged and elderly, and regions with lower SDI levels.

Self-harm and interpersonal violence due to high temperature from the global burden of disease study 2019: A 30-year assessment.

BACKGROUND: The impact of global warming on health due to climate change is increasingly studied, but the global burden of self-harm and interpersonal violence attributable to high temperature is still limited. This study aimed to systematically assess the burden of self-harm and interpersonal violence attributable to high temperature globally or by region and climate zone from 1990 to 2019. METHODS: We obtained the global, regional, and national deaths, disability-adjusted life years (DALYs), age-standardized mortality rates (ASMR), and age-standardized disability-adjusted life year rates (ASDR) of self-harm and interpersonal violence due to high temperature from 1990 to 2019 through the Global Burden of Disease Study (GBD) 2019. The burden of self-harm and interpersonal violence due to high temperature was estimated by age, sex, climate zone, the socio-demographic index (SDI), and the healthcare access and quality index (HAQ). Average annual percentage changes (AAPCs) in ASMR and ASDR were calculated for 1990-2019 using the Joinpoint model. RESULTS: From 1990 to 2019, the global deaths and DALYs related to self-harm and interpersonal violence due to high temperature increased from 20,002 (95% UI, 9243 to 41,928) and 1,107,216 (95% UI, 512,062 to 2,319,477) to 26,459 (95% UI, 13,574 to 47,265) and 1,382,487 (95% UI, 722,060 to 2,474,441), respectively. However, the ASMR and ASDR showed varying degrees of decreasing trends, with decreases of 13.36% and 12.66%, respectively. The ASMR was high and declining in low and low-middle SDI regions, particularly in tropical and subtropical regions. In addition, SDI and HAQ index were negatively correlated with ASMR in 204 countries and regions. CONCLUSIONS: The global burden of self-harm and interpersonal violence attributed to high temperature has decreased over the past 30 years, but the number of deaths and DALYs continues to rise. Climate change continues to make heat stress a significant risk factor for self-harm and interpersonal violence worldwide.

Metabolic profiles of Fufang Xiling Jiedu capsule in rats by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry.

Fufang Xiling Jiedu capsule (FXJC), a traditional Chinese medicine that evolved from "Yinqiao Powder", is widely used for the treatment of cold and influenza. However, due to a lack of in vivo metabolism research, the chemical components responsible for the therapeutic effects still remain unclear. Hence, this study aimed to describe the metabolic profiles of the FXJC in rat plasma, urine, and feces. A combined data mining strategy based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry was employed and 201 xenobiotics, including 117 prototype components and 84 metabolites were detected. Phenolic acids, flavonoids, triterpenes, and lignans were prominent ingredients absorbed in vivo, and the major metabolic pathways of the detected metabolites were glucuronidation, sulfation, methylation, and oxidation. This is the first systematic study on the metabolism of the FXJC in vivo, providing valuable information for future studies on the efficacy, toxicity, and mechanism of the FXJC.

The association between endocrine disrupting chemicals and MAFLD: Evidence from NHANES survey.

Previous studies on the association of endocrine-disrupting chemicals (EDCs) with metabolic dysfunction-associated fatty liver disease (MAFLD) are very limited. This study analyzed the association of EDCs exposure with MAFLD among 5073 American adults from the 2017-2018 National Health and Nutrition Examination Survey. The results showed that increased exposure to 3 EDCs metabolites (namely As, DiNP and PFOA) were significantly associated with MAFLD, the odds ratio of which were 1.819 (95% CI: 1.224, 2.702), 1.959 (95% CI: 1.224, 3.136) and 2.148 (95% CI: 1.036, 4.456), respectively. Further, the bayesian kernel machine regression model also revealed that phthalates exposure was strongly connected with the MAFLD, particularly in females and the elderly over 65. Moderating effect analysis suggested that higher body mass index (BMI) and inflammatory diet habit (indicated by dietary inflammatory index) strengthened the association between EDCs and MAFLD, whereas population with higher level of insulin sensitivity showed lower risk. In conclusion, our results suggest that either single or combined exposure to EDCs metabolites is link to MAFLD. Our findings also encourage people to sustain a healthy diet, normal levels of insulin sensitivity and BMI, which may help to alleviate the association of MAFLD risk in exposure to EDCs. These results also help us to better understand the association of EDCs and MAFLD and provide effective evidences for preventing MAFLD from the EDCs exposure aspect.

Machine-Learning-Based Calibration of Temperature Sensors.

Temperature sensors are widely used in industrial production and scientific research, and accurate temperature measurement is crucial for ensuring the quality and safety of production processes. To improve the accuracy and stability of temperature sensors, this paper proposed using an artificial neural network (ANN) model for calibration and explored the feasibility and effectiveness of using ANNs to calibrate temperature sensors. The experiment collected multiple sets of temperature data from standard temperature sensors in different environments and compared the calibration results of the ANN model, linear regression, and polynomial regression. The experimental results show that calibration using the ANN improved the accuracy of the temperature sensors. Compared with traditional linear regression and polynomial regression, the ANN model produced more accurate calibration. However, overfitting may occur due to a small sample size or a large amount of noise. Therefore, the key to improving calibration using the ANN model is to design reasonable training samples and adjust the model parameters. The results of this study are important for practical applications and provide reliable technical support for industrial production and scientific research.

Impact of different dietary fibre sources on production performance, bacterial composition and metabolites in the caecal contents of rabbits.

This study was conducted to investigate the effects of different dietary fibre sources on growth performance, gastrointestinal tract development, caecal fermentation and bacterial composition in the caecal contents of rabbits. A total of 120 35-day-old weaned Minxinan black rabbits were divided into three groups and fed a diet composed of peanut straw powder (Group A), alfalfa powder (Group B) and soybean straw powder (Group C) as the main fibre source. The final body weight and average daily gain in Group B were higher than those in Group C, and the average daily feed intake and feed conversion ratio in Group A were lower than those in Group C (p < 0.05). The relative weights of the stomach, small intestine and caecum of rabbits in Group C were higher than those in Groups of B and A, and the relative weights of the caecal contents in Group C were lower than those in Groups A or B (p < 0.05). The pH value and propionic acid, butyric acid and valeric acid concentrations in the caecum of Group C were lower than those in the caecum of Groups A or B, and the concentration of acetic acid in the caecum was lower (p < 0.05). The dominant microbes in the caecal contents of Minxinan black rabbits were Firmicutes, Bacteroidetes and Proteobacteria at the phylum level, and the number of species, Chao1 index and ACE index measured was different between the B-C and A-C groups (p < 0.05). Different dietary fibre sources could affect the growth performance, gastrointestinal tract development and intestinal microbiota of rabbits, and the nutritional value of alfalfa powder was better than that of peanut straw and soybean straw.

Water Activation Triggered by Cu-Co Double-Atom Catalyst for Silane Oxidation.

High-performance catalysts sufficient to significantly reduce the energy barrier of water activation are crucial in facilitating reactions that are restricted by water dissociation. Herein we present a Cu-Co double-atom catalyst (CuCo-DAC), which possesses a uniform and well-defined CuCoN6 (OH) structure, and works together to promote water activation in silane oxidation. The catalyst achieves superior catalytic performance far exceeding that of single-atom catalysts (SACs). Various functional silanes are converted into silanols with up to 98 % yield and 99 % selectivity. Kinetic studies show that the activation energy of silane oxidation by CuCo-DAC is significantly lower than that of Cu single-atom catalyst (Cu-SAC) and Co single-atom catalyst (Co-SAC). Theoretical calculations demonstrate two different reaction pathways where water splitting is the rate-determining step and it is accelerated by CuCo-DAC, whereas H2 formation is key for its single-atom counterpart.

Catalytic Activity Enhancement on Alcohol Dehydrogenation via Directing Reaction Pathways from Single- to Double-Atom Catalysis.

To further improve the intrinsic reactivity of single-atom catalysts (SACs), the controllable modification of a single site by coordinating with a second neighboring metal atom, developing double-atom catalysts (DACs), affords new opportunities. Here we report a catalyst that features two bonded Fe-Co double atoms, which is well represented by an FeCoN6(OH) ensemble with 100% metal dispersion, that work together to switch the reaction mechanism in alcohol dehydrogenation under oxidant-free conditions. Compared with Fe-SAC and Co-SAC, FeCo-DAC displays higher activity performance, yielding the desired products in up to 98% yields. Moreover, a broad diversity of benzyl alcohols and aliphatic alcohols convert into the corresponding dehydrogenated products with excellent yields and high selectivity. The kinetic reaction results show that lower activation energy is obtained by FeCo-DAC than that by Fe-SAC and Co-SAC. Moreover, computational studies demonstrate that the reaction path by DACs is different from that by SACs, providing a rationale for the observed enhancements.

Impact of high, low, and non-optimum temperatures on chronic kidney disease in a changing climate, 1990-2019: A global analysis.

BACKGROUND: Although a few studies have reported the relationship between high and low temperatures and chronic kidney disease (CKD), the global burden of CKD attributable to extreme heat and cold in recent decades remains unknown. METHODS: Based on the Global Burden of Disease Study (GBD) 2019, we obtained data on age-standardized mortality rates (ASMR) and age-standardized rates of disability-adjusted life years (ASDR) per 100 000 population of the CKD attributable to non-optimum temperatures from 1990 to 2019. The annual mean temperature of each country was used to divide each country into five climate zones (tropical, subtropical, warm-temperate, cool-temperate, and boreal). The locally weighted regression model was used to estimate the burden for different climate zones and Socio-demographic index (SDI) regions. RESULTS: In 1990, the ASMR and ASDR due to high temperature estimated -0.01 (95% UI, -0.74 to 0.44) and -0.32 (-21.66 to 12.66) per 100 000 population, respectively. In 2019, the ASMR and ASDR reached 0.10 (-0.28 to 0.38) and 2.71 (-8.07 to 10.46), respectively. The high-temperature burden increased most rapidly in tropical and low SDI regions. There were 0.99 (0.59 to 1.39) ASMR attributable to low-temperature in 1990, which increased to 1.05 (0.61-1.49) in 2019. While the ASDR due to low temperature declined from 22.03 (12.66 to 30.64) in 1990 to 20.43 (11.30 to 29.26) in 2019. Overall, the burden of CKD attributable to non-optimal temperatures has increased from 1990 to 2019. CKD due to hypertension and diabetes mellitus were the primary causes of CKD death attributable to non-optimum temperatures in 2019 with males and older adults being more susceptible to these temperatures. CONCLUSIONS: The CKD burden due to high, low, and non-optimum temperatures varies considerably by regions and countries. The burden of CKD attributable to high temperature has been increasing since 1990.

A global burden assessment of lung cancer attributed to residential radon exposure during 1990-2019.

This study aimed to explore the spatial and temporal trends of lung cancer burden attributable to residential radon exposure at the global, regional, and national levels. Based on the Global Burden of Disease Study (GBD) 2019, we collected the age-standardized mortality rate (ASMR) and age-standardized disability-adjusted life rate (ASDR) of lung cancer attributable to residential radon exposure from 1990 to 2019. The Joinpoint model was used to calculate the annual average percentage change (AAPC) to evaluate the trend of ASMR and ASDR from 1990 to 2019. The locally weighted regression (LOESS) was used to estimate the relationship of the socio-demographic index (SDI) with ASMR and ASDR. In 2019, the global ASMR and ASDR for lung cancer attributable to residential radon exposure were 1.03 (95% CI: 0.20, 2.00) and 22.66 (95% CI: 4.49, 43.94) per 100 000 population, which were 15.6% and 23.0% lower than in 1990, respectively. According to the estimation, we found the lung cancer burden attributable to residential radon exposure declined significantly in high and high-middle SDI regions, but substantially increased in middle and low-middle SDI regions from 1990 to 2019. Across age and sex, the highest burden of lung cancer attributable to residential radon exposure was found in males and elderly groups. In conclusion, the global burden of lung cancer attributable to residential radon exposure showed a declining trend from 1990 to 2019, but a relatively large increase was found in the middle SDI regions. In 2019, the burden of lung cancer attributable to residential radon exposure remained high, particularly in males, the elderly, and high-middle SDI regions compared with other groups.

Multioctave and reconfigurable frequency-stepped radar waveform generation based on an optical frequency shifting loop.

A photonic method for multioctave and reconfigurable frequency-stepped radar waveform generation is proposed and experimentally demonstrated based on an optical frequency shifting loop (OFSL). When a rectangular optical pulse is applied to the OFSL, a frequency-stepped optical signal can be generated. Beating the signal with another continuous-wave optical carrier, an electrical frequency-stepped waveform can be obtained. By meticulously adjusting the relations between the time duration of the rectangular optical pulse and the loop delay of the OFSL, the frequency-hopping rate (or the frequency-hopping period) of the generated frequency-stepped signal can be reconfigured. An experiment is carried out. The generation of frequency-stepped signals with frequency intervals of 1 GHz, 3 GHz, 5 GHz, 8 GHz, and 10 GHz is realized. The reconfigurability of the frequency-hopping period is also investigated and different frequency-hopping periods of 189, 10.2, 5.1, and 2.42 ns are achieved.

Discs large homolog 1 regulates B-cell proliferation and antibody production.

Antibody production results from B-cell activation and proliferation upon antigen binding. Discs large homolog 1 (Dlg1), a scaffold protein from the membrane-associated guanylate kinase family, has been shown to regulate the antigen receptor signaling and cell polarity in lymphocytes; however, the physiological function of Dlg1 in humoral responses is not completely clear. Here, we addressed this question using a conditional knockout (KO) mouse model with Dlg1 deficiency in different B-cell subsets by crossing dlg1fl/fl mice with either mb1cre/+ or aicdacre/+ mice, respectively. In both mouse models, we observed that Dlg1 deficiency in B cells (Dlg1-KO B cells) led to obvious hyper-antibody responses upon immunization, the effect of which was more obvious in antigen-recall responses. Mechanistically, we found that Dlg1-KO B cells exhibited hyper-proliferation compared with wild-type B cells upon antigen stimulation, suggesting that the hyper-antibody responses are likely induced by the hyper-proliferation of Dlg1-KO B cells. Indeed, further studies demonstrated that Dlg1 deficiency in B cells led to the down-regulation of a tumor suppressor, FoxO1. Thus, all these results reveal an unexpected function of Dlg1 in restraining hyper-antibody responses through the inhibition of FoxO1 and thus antigen-binding-induced proliferation in B cells.

Copper-Catalyzed Three-Component Redox-Neutral Ring Opening of Benzothiazoles to 1-Amino-N-(2-(phenylthio)phenyl)methanimine.

Copper-catalyzed three-component redox-neutral ring opening of benzothiazoles with aryl iodides and O-benzoyl hydroxylamines for the synthesis of 1-amino-N-(2-(phenylthio)phenyl)methanimine has been developed. This one-pot reaction undergoes C-S and N-O bond cleavage and new C-S and C-N bond construction. Several control experiments excluded a free radical procedure and also demonstrated the secondary amine as a possible intermediate, which was vital to the catalytic reaction. Meanwhile, the deuteration experiment got rid of the C-H activation dehydroisomerization of the benzothiazole mechanism.

Palladium-Catalyzed Amination/Dearomatization Reaction of Indoles and Benzofurans.

This report describes a palladium-catalyzed dearomatization and amination tandem reaction of 2,3-disubstituted indoles and benzofurans via the Catellani strategy. This reaction provides a new method for the construction of amino-substituted indoline-fused cyclic and benzofuran spiro compounds in good yields. The reaction has broad functional group compatibility and substrate scope.

Palladium-Catalyzed ortho-C-H Glycosylation/ipso-Alkenylation of Aryl Iodides.

This report describes the first example of palladium-catalyzed ortho-C-H glycosylation/ipso-alkenylation of aryl iodides, and the easily accessible glycosyl chlorides are used as a glycosylation reagent. The reaction is compatible with the functional groups of the substrates, and a series of C-aryl glycosides have been synthesized in good to excellent yield and with excellent diastereoselectivity. It is found that a cheap 5-norbornene-2-carbonitrile as a transient mediator can effectively promote this reaction. In addition, ipso-arylation and cyanation were also realized by the strategy.

Triplet Male Lambs Are More Susceptible than Twins to Dietary Soybean Oil-Induced Fatty Liver.

BACKGROUND: Litter size affects fetal development but its relation to diet-induced fatty liver later in life is unknown. OBJECTIVES: This aim of this study was to test the hypothesis that litter size influences postweaning fatty liver development in response to soybean oil-supplemented diet. METHODS: Weanling twin (TW) or triplet (TP) male lambs (n = 16) were fed a control diet or 2% soybean oil-supplemented diet (SO) for 90 d. Liver tissue morphology, biochemical parameters, and lipid metabolic enzymes were determined. Hepatic gene expression was analyzed by RNA sequencing (n = 3), followed by enrichment analysis according to Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes. Differentially expressed genes involved in lipid metabolism were further verified by quantitative reverse transcriptase-polymerase chain reaction (n = 4). All data were analyzed by a 2-factor ANOVA, apart from differentially expressed genes, which were identified by the Benjamini-Hochberg approach (q value ≤0.05). RESULTS: SO increased liver triglyceride (by 55%) and nonesterified fatty acid (by 54%) concentrations in TPs (P ≤ 0.05) but not in TWs (P > 0.05). SO also induced a 2.3- and 2.1-fold increase in the liver steatosis score of TPs and TWs, respectively (P ≤ 0.05). Moreover, SO reduced the activity of lipolytic enzymes including hepatic lipase and total lipase in TPs by 47% and 25%, respectively (P ≤ 0.05). In contrast, activities of lipogenic enzymes, including malic enzyme and acetyl coenzyme A carboxylase, were significantly higher in TPs (P ≤ 0.05). Moreover, TPs had higher expression of lipogenic genes, such as FASN (by 45%) and APOB (by 72%), and lower expression of lipolytic genes, such as PRKAA2 (by 28%) and CPT1A (by 43%), compared with TWs (P ≤ 0.05). CONCLUSIONS: TPs have a gene expression profile that is more susceptible to SO-induced fatty liver than that of TWs, which indicates that insufficient maternal nutrient supply at fetal and neonatal stages may increase the risk of nonalcoholic fatty liver disease.