Regional Variations in Physical Fitness in Children and Adolescents in Shaanxi Province.

Objectives: This study aims to examine the geographical variation in physical fitness levels among Chinese children and adolescents in Shaanxi province. Methods: A total of 19,175 children from Shaanxi province with physical fitness data in 2019, participated in the study. Physical fitness was assessed using body mass index, force vital capacity, 50 m sprint, sit and reach, 1 min rope skipping, sit-ups, 50 m × 8 round-trip running, standing long jump, pull-ups, 800 m, and 1000 m running, and their standardized scores were aggregated to form a summary score. The total score is used to classify the physical fitness levels into four grades (excellence to failure). Results: The Guanzhong (GZ) region scored the highest, while Northern Shaanxi (NS) scored the lowest. The excellence rate for physical fitness was highest in GZ and lowest in NS, while the failure rate was highest in NS and lowest in GZ. Notably, children and adolescents in NS demonstrated the best endurance levels despite their overall lower fitness scores. The comprehensive physical fitness among Chinese children and adolescents in Shaanxi province showed significant regional disparities. GZ region exhibited the highest physical fitness levels, while Northern Shaanxi had the lowest. Conclusions: Region-specific interventions and targeted health policies are essential to address these disparities and improve the overall physical health status of children and adolescents in Shaanxi province.

Variability in the home-field advantage of litter decomposition mediates alterations in soil CO2 and CH4 fluxes: A transplantation experiment study.

The decomposition of litter is susceptible to the influence of climate change and soil conditions, which can subsequently impact the release of carbon dioxide (CO2) from forest soils and the absorption of methane (CH4). Ecological theory proposes the existence of a home-field advantage (HFA) in litter decomposition, suggesting that the decomposition rate of litter (such as fallen leaves, twigs, and roots) may be faster in their native habitat than in foreign environments. Therefore, we selected litter from Pinus tabuliformis (PT) and Quercus acutissima Carruth (QC) in the field and conducted a 439-day litter transplant experiment to test the magnitude and direction of the HFA of these two litter types in three forest stands. During this experiment, we monitored the changes in soil CO2 and CH4 fluxes associated with the decomposition of PT and QC leaf litter in their native and foreign sites. Furthermore, we measured various soil physical, chemical, and biological indicators. The results indicated that the decomposition rate of QC leaf litter was faster in its native habitat, demonstrating a clear HFA effect. Conversely, the decomposition of PT leaf litter was observed to be more rapid in away soil, suggesting a pronounced home-field disadvantage (HFD). The study found that PT leaf litter exhibited greater CO2 release when decomposing in away soil, demonstrating 43 % and 32 % increases compared to bare soil, respectively. Conversely, QC leaf litter was observed to release more CO2 in its home soil. Additionally, the bare soils of the PT and QC home sites were found to absorb more CH4 compared to leaf litter coverage, with increases of 37.8 % and 31.2 %, respectively. The partial least squares model indicated that the litter attributes had a significant direct effect on soil temperature and enzyme activity. Soil temperature and enzyme activity further directly influenced the soil CO2 and CH4 fluxes. The results of our study indicate that the HFA of litter is dependent on litter type, and that litter transplantation can impact soil greenhouse gas exchange. This research provides a theoretical foundation for forest management and conservation strategies, as well as valuable data for global carbon neutrality efforts.

Experimental Study of the Flexural Performance of GFRP-Reinforced Seawater Sea Sand Concrete Beams with Built-In GFRP Tubes.

The use of seawater sea sand concrete (SSSC) and fiber-reinforced polymer (FRP) has broad application prospect in island and coastal areas. However, the elastic modulus of FRP reinforcement is obviously lower than that of ordinary steel reinforcement, and the properties of SSSC are different from that of ordinary concrete, which results in a limit in the bearing capacity and stiffness of structures. In order to improve the flexural performance of FRP-reinforced SSSC beams, a novel SSSC beam with built-in glass FRP (GFRP) tubes was proposed in this study. Referring to many large-scale beam experiments, one specimen was used for one situation to illustrate the study considering costs and feasibility. Firstly, flexural performance tests of SSSC beams with GFRP tubes were conducted. Then, the effects of the GFRP tubes' height, the strength grades of concrete inside and outside the GFRP tubes, and the GFRP reinforcement ratio on the flexural behaviors of the beams were investigated. In addition, the concept of capacity reserve was proposed to assess the ductility of the beams, and the interaction between the concrete outside the GFRP tube, the GFRP tube and concrete inside the tube was discussed. Finally, the formulas for the normal section bearing capacity of beams with built-in GFRP tubes were derived and verified. Compared to the beam without GFRP tubes, under the same conditions, the ultimate bearing capacities of the SSSC beam with 80 mm, 100, and 200 mm height GFRP tubes were increased by 17.67 kN, 24.52 kN, and 144.42 kN, respectively.

Determination of Azole Fungicide Residues in Fresh Juice by Magnetic Solid Phase Extraction Based on Fe3O4@ZnAl-LDH@MIL-53(Al) Sorbent in Combination with High-Performance Liquid Chromatograph.

In this work, a magnetic adsorption material based on metal-organic framework (Fe3O4@ZnAl-LDH@MIL-53(Al)) was synthesized and used as an adsorbent in the process of magnetic solid phase extraction. Then, a high-performance liquid chromatograph was used to quantitatively detect triazole fungicides in samples. In order to verify the successful preparation of the material, a series of characterization analyses were carried out. Besides, the key parameters that may affect the extraction efficiency have been optimized, and under optimal conditions the three triazole fungicides showed good linearity in the range of 10-1000 µg/L (R2 ≥ 0.9796); Limit of detections were ranged from 0.013 to 0.030 µg/mL. Finally, the established method was applied to the detection of triazole fungicides in four fresh juice samples. The results showed that the target analyte was not detected in all the test samples. By detecting the recoveries (73.3-104.3%) and coefficient variation (RSD ≤ 6.8%) of triazole fungicides in fortified samples, it proved that this established method meets the requirements of pesticide residue analysis and showed excellent application potential.

Study on Capillary Water Absorption of Waterborne-Polyurethane-Modified Recycled Coarse Aggregate Concrete.

The reuse of construction and demolition waste as a substitute for natural coarse aggregate in the production of recycled concrete has been widely used. In order to study the capillary water absorption performance of waterborne-polyurethane-modified recycled aggregate concrete (WPUMRC), the effects of different curing systems, polymer-cement ratios, and waterborne polyurethane addition methods on the cumulative water absorption and the rate of capillary water absorption of WPUMRC were analyzed, and through MIP tests, the micro modification mechanism of waterborne polyurethane in recycled concrete was analyzed. The results indicate that the optimal curing system for both DC (waterborne polyurethane is added separately from water) and HC (waterborne polyurethane is mixed with some effective water and then added) is the 14 d standard curing-14 d indoor natural drying curing system. Waterborne polyurethane can fill the pores and micro-cracks inside WPUMRC or interweave with the hydration products of cement to form a spatial network structure, reducing the porosity, and thereby improving the capillary water absorption performance of WPUMRC. Based on the MIP test results, the grey correlation method was used to establish the relationship between capillary water absorption and the pore structure of WPUMRC under the optimal curing system. In addition, the prediction model of capillary water absorption in recycled concrete was established according to the test results, which can be used to predict WPUMRC's capillary water absorption performance.

Multifunctional Oxidized Dextran-Metformin as a Tissue-Adhesive Hydrogel to Prevent Postoperative Peritoneal Adhesions in Patients with Metabolic Syndrome.

Patients with metabolic syndrome (MetS) undergoing surgery are at high risk of developing peritoneal adhesions and other severe postoperative complications. However, the single shielding function and absence of physiological activity render conventional methods less useful in preventing adhesions in patients with MetS. To address this challenge, a convenient method is introduced for developing a novel tissue-adhesive hydrogel called oxidized dextran-metformin (ODE-ME) via Schiff base linkages. This injectable ODE-ME hydrogel exhibits excellent tissue-adhesive properties and various physiological functions, particularly enhanced antibacterial effects. Furthermore, in vivo experiments demonstrate that the hydrogel can effectively alleviate hyperglycemia, reduce excessive inflammation, and improve fibrinolytic activity in MetS mice, thereby preventing adhesions and promoting incisional healing. The hydrogel concurrently isolates injured tissues and lowers the blood glucose levels immediately after surgery in mice. Therefore, the ODE-ME hydrogel functions as a multifunctional barrier material and has potential for preventing postoperative peritoneal adhesions in patients with MetS in clinical settings.

Effects of a Ganoderma lucidum Proteoglycan on Type 2 Diabetic Rats and the Recovery of Rat Pancreatic Islets.

Type 2 diabetes (T2D) results from both insulin resistance and pancreatic ß-cell dysfunction. A natural proteoglycan extracted from Ganoderma lucidum, namely, FYGL, has been demonstrated to be capable of ameliorating insulin resistance in previous work. In this work, a T2D rat model induced by streptozocin (STZ) and a high-fat diet was used to investigate the effects of FYGL on pancreatic functions, and the transcriptomics of the rat pancreas was used to investigate the biological processes (BP) and signal pathways influenced by FYGL on the gene basis. Furthermore, the results of transcriptomics were verified both by histopathological analyses and protein expression. The studies showed that FYGL positively regulated T2D-related BP and signaling pathways and recovered the pancreatic function, therefore ameliorating hyperglycemia and hyperlipidemia in vivo. Importantly, the recovery of the pancreatic function suggested a crucial strategy to radically treat T2D.

Pitaya Virus X Coat Protein Acts as an RNA Silencing Suppressor and Can Be Used as a Specific Target for Detection Using RT-LAMP.

Selenicereus undatus (Haworth) D.R. Hunt (pitaya) is a tropical fruit that has been commonly cultivated in Guizhou Province, China, in recent years due to its good taste and high nutritional value. This planting area currently ranks third in China. Viral diseases have increasingly emerged in pitaya cultivation because of the expansion of the pitaya planting area and the characteristics of asexual propagation. The spread of pitaya virus X (PiVX; a Potexvirus) is among the most severe viruses threatening the quality and yield of pitaya fruit. To investigate the occurrence of PiVX in pitaya cultivations in Guizhou Province, we developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method that can detect PiVX with high sensitivity and specificity at a low cost and produce a visualized result. Our best RT-LAMP system was significantly more sensitive than RT-PCR and was highly specific to PiVX. Furthermore, PiVX coat protein (CP) can form a homodimer, and PiVX may use its CP as a plant RNA silencing suppressor to enhance infection. To the best of our knowledge, this is the first report of fast detection of PiVX and functional exploration of CP in a Potexvirus. These findings will provide an opportunity for early diagnosis and prevention of viruses in pitaya.

One-Step Construction of Tryptophan-Derived Small Molecule Hydrogels for Antibacterial Materials.

Amino acid-based hydrogels have received widespread attention because of their wide range of sources, biodegradability, and biocompatibility. Despite considerable progress, the development of such hydrogels has been limited by critical problems such as bacterial infection and complex preparation. Herein, by using the non-toxic gluconolactone (GDL) to adjust the pH of the solution to induce the rapid self-assembly of N-[(benzyloxy)carbonyl]-L-tryptophan (ZW) to form a three-dimensional (3D) gel network, we developed a stable and effective self-assembled small-molecule hydrogel. Characterization assays and molecular dynamics studies indicate that π-π stacking and hydrogen bonding are the main drivers of self-assembly between ZW molecules. In vitro experiments further confirmed this material's sustained release properties, low cytotoxicity, and excellent antibacterial activity, particularly against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. This study provides a different and innovative perspective for the further development of antibacterial materials based on amino acid derivatives.

Ternary complexes of cyclodextrins with alkali earth cations and amino acids in gas phase investigated by mass spectrometry.

Noncovalent ternary complexes between cyclodextrins (CDs), small molecules and alkali earth cations drew growing attention due to their potential application in many chemical and pharmaceutical fields. To date, the main factors affect the formation mechanism of noncovalent ternary complexes in gas phase have not been fully investigated. In this study, ternary complexes of CDs, divalent metal cations and amino acids (AAs) were investigated by electrospray ionization mass spectrometry (ESI-MS), demonstrating the formation of 1:1:1 stoichiometric noncovalent ternary complex of [CD + cation(II)+AA]2+ in gas phase. The results revealed that only +2 valence cations can form stable ternary complexes in ESI-MS. The ratio of peak intensities for [ß-CD + Mg(II)+AA]2+ to those for [ß-CD + Mg(II)]2+ hydrophobicity of AAs was also determined to discuss the effect of hydrophobicity of AAs. Exceptions exist for Pro, Gly, and Val indicated that other factors such as side-chain structure and rigidity of AAs can also influence the binding strength for ternary complexes. Collision induced dissociations (CID) were performed to further confirm the formation of the ß-CD ternary complexes, revealing the binding strength of [CD + Mg(II)+Phe]2+ decreased in the order of γ-CD, ß-CD, and α-CD. Although Leu and Ile are isomers, the ESI-MS demonstrated the peak intensity for ternary complexe of [ß-CD + Mg(II)+Ile]2+ exhibited stronger than that of [ß-CD + Mg(II)+Leu]2+, DFT theoretical calculations were conducted to explain the phenomenon. The calculation indicated when Mg2+ existing, the conformations of the two ternary complexes could be affected due to the electrostatic force. In the complexes, the Leu and Ile turn a way round, inserting to the cavity with their carboxylic acid side into the large rim side of ß-CD and interacting with Mg2+. This work not only clearly explained the factors influencing the formation of [CD + cation(II)+AA]2+ in gas phase, but it also provides an insight in designing ternary complexes for areas such as drug design and chiral discrimination.

Integrated transcriptome analysis reveals roles of long non-coding RNAs (lncRNAs) in caprine skeletal muscle mass and meat quality.

Long non-coding RNAs (lncRNAs) play important roles in the growth and development of skeletal muscle. However, there is limited information on goats. In this study, expression profiles of lncRNAs in Longissimus dorsi muscle from Liaoning cashmere (LC) goats and Ziwuling black (ZB) goats with divergent meat yield and meat quality were compared using RNA-sequencing. Based on our previous microRNA (miRNA) and mRNA profiles obtained from the same tissues, the target genes and binding miRNAs of differentially expressed lncRNAs were obtained. Subsequently, lncRNA-mRNA interaction networks and a ceRNA network of lncRNA-miRNA-mRNA were constructed. A total of 136 differentially expressed lncRNAs were identified between the two breeds. Fifteen cis target genes and 143 trans target genes were found for differentially expressed lncRNAs, and they were enriched in muscle contraction, muscle system process, muscle cell differentiation, and p53 signaling pathway. A total of 69 lncRNA-trans target gene pairs were constructed, with close relationship with muscle development, intramuscular fat deposition, and meat tenderness. A total of 16 lncRNA-miRNA-mRNA ceRNA pairs were identified, of which some reportedly associated with skeletal muscle development and fat deposition were found. The study will provide an improved understanding of the roles of lncRNAs in caprine meat yield and meat quality.

Deep Small RNA Sequencing Reveals Important miRNAs Related to Muscle Development and Intramuscular Fat Deposition in Longissimus dorsi Muscle From Different Goat Breeds.

MicroRNAs (miRNAs) are a class of small non-coding RNAs that have been shown to play important post-transcriptional regulatory roles in the growth and development of skeletal muscle tissues. However, limited research into the effect of miRNAs on muscle development in goats has been reported. In this study, Liaoning cashmere (LC) goats and Ziwuling black (ZB) goats with significant phenotype difference in meat production performance were selected and the difference in Longissimus dorsi muscle tissue expression profile of miRNAs between the two goat breeds was then compared using small RNA sequencing. A total of 1,623 miRNAs were identified in Longissimus dorsi muscle tissues of the two goat breeds, including 410 known caprine miRNAs, 928 known species-conserved miRNAs and 285 novel miRNAs. Of these, 1,142 were co-expressed in both breeds, while 230 and 251 miRNAs were only expressed in LC and ZB goats, respectively. Compared with ZB goats, 24 up-regulated miRNAs and 135 miRNAs down-regulated were screened in LC goats. A miRNA-mRNA interaction network showed that the differentially expressed miRNAs would target important functional genes associated with muscle development and intramuscular fat deposition. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that the target genes of differentially expressed miRNAs were significantly enriched in Ras, Rap 1, FoxO, and Hippo signaling pathways. This study suggested that these differentially expressed miRNAs may be responsible for the phenotype differences in meat production performance between the two goat breeds, thereby providing an improved understanding of the roles of miRNAs in muscle tissue of goats.

The chirality determination of amino acids by forming complexes with cyclodextrins and metal ions using ion mobility spectrometry, and a DFT calculation.

Chiral recognition is of highly interest in the areas of chemistry, pharmaceuticals, and bioscience. An effective strategy of enantiomeric determination of amino acids (AAs) was developed in this work. All 19 natural AAs enantiomers can be easily distinguished by ion mobility-mass spectrometry of the non-covalent complexes of AAs with cyclodextrins (α-CD, ß-CD and γ-CD) and Mg2+ without any chemical derivatization. Differences of the mobilities between the enantiomers' complexes is from 0.006 to 0.058 V s/cm2. In addition, the complex of [ß-CD + Phe + Mg]2+ was selected as an example to study the relative quantification by measuring L/D-Phe at different molar ratio of 10:1 to 1:10 in the µM range, resulting in a good linearity (R2 > 0.99) and high sensitivity at 2 µM. A DFT calculation was also performed to illustrate the detailed molecular structure of the complexes of CDs, Mg2+ and D- or L-Phe. Both experiment and theoretical calculation showed that Mg2+ plays an important role in host/guest interactions, which changed the molecular conformations by non-covalent interaction between Mg2+ and CDs, and resulted in the different collision cross-sections of the complex ions of CDs, Mg2+ and D- or L-AAs in the gas phase. This effective and convenient strategy could potentially be utilized in scientific research and industry for routine enantiomeric determination of natural AAs, peptides and some other small chiral biomolecules such as non-natural AAs and carboxylic acid-containing drugs.

Effects of sleep deprivation of various durations on novelty-related object recognition memory and object location memory in mice.

Sleep is essential for important physiological functions. Impairment of learning and memory function caused by lack of sleep is a common physiological phenomenon of which underlying changes in synaptic plasticity in the hippocampus are not well understood. The possible different effects of sleep deprivation (SD) lasting for various durations on learning and memory function and hippocampal synaptic plasticity are still not completely clear. In this study, we used a modified multiple platform method (MMPM) to induce rapid eye movement SD (REM SD), lasting for 24 h, 48 h, and 72 h, separately. The novel place recognition (NPR) and novel object recognition (NOR) tasks were used to test the novelty-related object recognition memory (ORM) and object location memory (OLM) of mice. Then, hippocampal synaptic plasticity was evaluated after all behavioural experiments. The results showed that REM SD played a key role in OLM but not in ORM. Specifically, 24 h REM SD improved novelty-related OLM, accompanied by a significantly increased hippocampal synaptic plasticity, including gain of dendritic spines, increased expression of hippocampal GluA1, and enhanced long-term potentiation (LTP), whereas 48 h REM SD showed no effect on OLM or the hippocampal synaptic plasticity mentioned above; however, 72 h REM SD impaired novelty-related OLM and weakened hippocampal synaptic plasticity, including serious loss of dendritic spines, decreased expression of hippocampal GluA1, and significantly attenuated LTP. Our results suggest that REM SD of various durations has different effects on both novelty-related OLM and hippocampal synaptic plasticity.

The thermodynamic and kinetic mechanisms of a Ganoderma lucidum proteoglycan inhibiting hIAPP amyloidosis.

Ganoderma lucidum is a valuable medicinal herbal which has been reported to prevent type 2 diabetes (T2D). A natural hyperbranched proteoglycan extracted from Ganoderma lucidum, namely, FYGL, has been demonstrated to inhibit the amyloidosis of human islet amyloid polypeptide (hIAPP) previously by our lab. However, the effective active components and the mechanisms of FYGL in inhibiting hIAPP amyloidosis are unknown. To identify the effective active components, different components from FYGL were isolated: the polysaccharide FYGL-1, the proteoglycans of FYGL-2 and FYGL-3. We further separated and sequenced the protein moieties of FYGL-2 and FYGL-3, namely, FYGL-2-P and FYGL-3-P, respectively, and compared their abilities to inhibit hIAPP amyloidosis, and systematically explored the inhibitory mechanisms by spectroscopy, microscopy and molecular dynamic simulation methods. Results showed that the protein moieties of FYGL played essential roles in inhibiting hIAPP amyloidosis. The strong, specific, and enthalpy-driven interaction by π-π stacking and electrostatic forces between hIAPP and FYGL-3-P dramatically inhibited hIAPP amyloidosis. These results suggested that FYGL-3-P had enormous potential to prevent hIAPP misfolding-induced diabetes and structurally helped researchers to seek or design inhibitors against polypeptide amyloidosis.

Interaction and Inhibition of a Ganoderma lucidum Proteoglycan on PTP1B Activity for Anti-diabetes.

Protein tyrosine phosphatase 1B (PTP1B) is a key negative regulator of insulin and an effective target for the treatment of type 2 diabetes (T2D). A natural hyperbranched proteoglycan extracted from Ganoderma lucidum, namely, Fudan-Yueyang G. Lucidum (FYGL), was demonstrated capable of inhibiting the activity of PTP1B. Here, to identify the effective active components of FYGL, three different components, the polysaccharide FYGL-1, proteoglycans FYGL-2, and FYGL-3, were isolated from FYGL, and then, the protein moiety of FYGL-3 was further separated, namely, FYGL-3-P. Their abilities to enhance the glucose uptake in cells and inhibit the activity of PTP1B were compared. The inhibitory mechanisms were systematically explored by spectroscopic methods and MD simulations. The results showed that FYGL-3 and FYGL-3-P significantly enhanced the insulin-provoked glucose uptake in insulin-resistant HepG2 cells, detected by the glucose oxidase method. Also, the FYGL-3-P protein moiety in FYGL played an essential role in inhibiting the activity of PTP1B. A strong, enthalpy-driven, and multitargeted interaction by electrostatic forces between PTP1B and FYGL-3-P dramatically inhibited the catalytic activity of PTP1B. These results provided deep insights into the molecular mechanisms of FYGL inhibiting the activity of PTP1B and structurally helped researchers seek natural PTP1B inhibitors.

Distinction of chiral penicillamine using metal-ion coupled cyclodextrin complex as chiral selector by trapped ion mobility-mass spectrometry and a structure investigation of the complexes.

Penicillamine (Pen) is a common chiral drug that is obtained from penicillin. Between the two enantiomers of Pen, only D-Pen can be used to treat cystinuria and rheumatoid arthritis while L-Pen is toxic. Therefore, it requires great efforts for the research of the rigorous analysis and distinction of the two enantiomers. The non-covalent combination of chiral molecules and chiral selectors (CSs) has been proved as a unique strategy for chiral distinction by ion mobility spectrometry in coupling with -mss spectrometry (IM-MS). Here, we developed a simple method to distinguish D, L-Pen by using special CSs for IM-MS separation. The CSs utilized here include cyclodextrins (CD) and linear chain oligosaccharides plus metal ions. We found that non-covalent complexes [Pen+ß-CD + Li]+ could be easily formed by electrospray ionization of the mixture of the solution, and the chirality of Pen could be effectively recognized by measuring their mobilities due to the different collision cross collision sections of [D-Pen+ß-CD + Li]+ and [L-Pen+ß-CD + Li]+. A detailed analysis of [Pen+ß-CD + Li]+ was then conducted by the optical rotation measurements and NMR experiments to reveal their structural differences. Furthermore, DFT calculation showed the differences of molecular conformation between the complexes. The results provide a new powerful method for fast analysis and recognition of chirality of Pen compounds by IM-MS.

Direct distinction of ibuprofen and flurbiprofen enantiomers by ion mobility mass spectrometry of their ternary complexes with metal cations and cyclodextrins in the gas phase.

Enantiomeric drugs are widely used and play important roles in pharmaceuticals. Ion mobility spectrometry coupled with mass spectrometry technology provides a unique method for distinguishing the enantiomeric drugs, enantiomeric identification, and quantitation in the gas phase. In this study, enantiomeric molecules of ibuprofen and flurbiprofen were clearly recognized by forming host-guest complex ions using trapped ion mobility time-of-flight mass spectrometry. Ternary complex ions can be produced easily by electrospray ionization of the mixed solutions of ibuprofen, cyclodextrins, and CaCl2 , LiCl, or NaCl, as well as flurbiprofen, cyclodextrins, and CaCl2 . The relative contents of different chiral ibuprofens in a mixed solution were also quantitatively measured. This new method is a simple, effective, and a convenient enantioselective analysis method.

Circulating mitochondrial DNA is a proinflammatory DAMP in sickle cell disease.

The pathophysiology of sickle cell disease (SCD) is driven by chronic inflammation fueled by damage associated molecular patterns (DAMPs). We show that elevated cell-free DNA (cfDNA) in patients with SCD is not just a prognostic biomarker, it also contributes to the pathological inflammation. Within the elevated cfDNA, patients with SCD had a significantly higher ratio of cell-free mitochondrial DNA (cf-mtDNA)/cell-free nuclear DNA compared with healthy controls. Additionally, mitochondrial DNA in patient samples showed significantly disproportionately increased hypomethylation compared with healthy controls, and it was increased further in crises compared with steady-state. Using flow cytometry, structured illumination microscopy, and electron microscopy, we showed that circulating SCD red blood cells abnormally retained their mitochondria and, thus, are likely to be the source of the elevated cf-mtDNA in patients with SCD. Patient plasma containing high levels of cf-mtDNA triggered the formation of neutrophil extracellular traps (NETs) that was substantially reduced by inhibition of TANK-binding kinase 1, implicating activation of the cGAS-STING pathway. cf-mtDNA is an erythrocytic DAMP, highlighting an underappreciated role for mitochondria in sickle pathology. These trials were registered at www.clinicaltrials.gov as #NCT00081523, #NCT03049475, and #NCT00047996.

Direct and simultaneous recognition of the positional isomers of aminobenzenesulfonic acid by TIMS-TOF-MS.

Positional isomer recognition is a challenging scientific issue. Fast and accurate detection of isomers is required for understanding their chemical properties. Here, we describe a method for simultaneous recognition of three positional isomers of 2-aminobenzenesulfonic acid (2-ABSA), 3-ABSA, and 4-ABSA using trapped ion mobility spectroscopy-time-of-flight mass spectrometry (TIMS-TOF-MS). The three ABSA positional isomers were recognized by measuring the different ion mobility of the ternary complexes of [ß-cyclodextrin (CD)+ABSA + Li]+ or [λ-CD + ABSA + Na]+, because their different collision cross-sections or different spatial conformations. The collision-induced dissociation mechanism of the different complexes of [ß-CD + ABSA + Li]+ and [λ-CD + ABSA + Na]+ using tandem mass spectrometry exhibited the same dissociation process with slightly different dissociation energies, which the smaller cross-section requires higher collision energy that means the smaller complex with tighter and more stable conformation than a larger complex for the ABSA complexes. In addition, relative quantification of the ABSA isomers was studied by measuring any two of the three ABSA isomer complexes at different molar ratio of 10:1 to 1:10 in the µM range, good linearity (R2 > 0.99) with precision between 2.14% and 2.58%, and accuracy ≥ 97.1% were obtained. The method for fast determination and recognition of ABSA positional isomers by combination with CD and alkali metal ions possesses the advantages of being simple, direct, rapid, sensitive, cost-effective, and needs no chemical derivatives or chromatographic separation before analysis. Therefore, the proposed method would be a powerful tool for the analysis of ABSA isomers or even other positional isomers.