A tough, reversible and highly sensitive humidity actuator based on cellulose nanofiber films by intercalation modulated plasticization.

Cellulose nanofiber was an ideal candidate for humidity actuators based on its wide availability, biocompatibility and excellent hydrophilicity. However, conventional cellulose nanofiber-based actuators faced challenges like poor water resistance, flexibility, and sensitivity. Herein, water-resistant, flexible, and highly sensitive cross-linked cellulose nanofibers (CCNF) single-layer humidity actuators with remarkable reversible humidity responsiveness were prepared by combining the green click chemistry modification and intercalation modulated plasticization (IMP). The incorporation of phenyl ring and the crosslinked network structure in CCNF films contributed to its improved water resistance and mechanical properties (with a stress increased from 85.9 ± 3.1 MPa to 141.2 ± 21.5 MPa). SEM analysis confirmed enhanced interlaminar sliding properties facilitated by IMP. This resulted in increased flexibility and toughness of CCNF films, with a strain of 11.5 % and toughness of 9.9 MJ/m3. These improvements efficiently enhanced humidity sensitivity for cellulose nanofiber, with a 4.8-fold increase in bending curvature and a response time of only 3.4 ± 0.1 s. Finally, the good humidity sensitivity of modified CNF can be easily imparted to carbon nanotubes (CNTs) via simple self-assembly method, thus leading to a high-performance humidity-responsive actuator. The click chemistry modification and IMP offer a new avenue to fabricate tough, reversible and highly sensitive humidity actuator based on cellulose nanofiber.

Investigation on the Correlation between Biaxial Stretching Process and Macroscopic Properties of BOPA6 Film.

Biaxially oriented polyamide 6 (BOPA6) films were prepared by extrusion casting and biaxial stretching with polyamide 6. The effects of different biaxially oriented on the macroscopic properties of BOPA6 were investigated by characterizing the rheological, crystallization, optical, barrier and mechanical properties. The results show that the increase of stretching temperature leads to the diffusion and regular stacking rate of BOPA6 chain segments towards crystal nuclei increases, the relative crystallinity increases, reaching 27.87% at 180 °C, and the mechanical strength and optical performance decrease. Heat-induced crystallization promotes the transformation of ß-crystals to α-crystals in BOPA6, resulting in a more perfect crystalline structure and enhancing oxygen barrier properties. BOPA6 chains are oriented, and strain-induced crystallization (SIC) occurs during the biaxial stretching. Further increasing the stretch ratio, the relative crystallinity increased to 30.34%. The machine direction (MD) and transverse direction (TD) tensile strength of BOPA6 (B-33) are nearly two times higher than the unstretched film, reaching 134.33 MPa and 155.28 MPa, respectively. In addition, the permeation decreases to 57.61 cc·mil/(m2 day), and the oxygen barrier performance has improved by nearly 30% compared to the sample B-22. BOPA6 has a high storage modulus at a high stretching rate (300%/s). Rapid chain relaxation would promote the molecular chain disorientation, destroy the entangled network of the molecular chain, and lead to a decrease in tensile strength, reducing to about 110 MPa.

Transcriptional patterns of human retinal pigment epithelial cells under protracted high glucose.

BACKGROUND: The retinal pigment epithelium (RPE) is essential for retinal homeostasis. Comprehensively exploring the transcriptional patterns of diabetic human RPE promotes the understanding of diabetic retinopathy (DR). METHODS AND RESULTS: A total of 4125 differentially expressed genes (DEGs) were screened out from the human primary RPE cells subjected to prolonged high glucose (HG). The subsequent bioinformatics analysis is divided into 3 steps. In Step 1, 21 genes were revealed by intersecting the enriched genes from the KEGG, WIKI, and Reactome databases. In Step 2, WGCNA was applied and intersected with the DEGs. Further intersection based on the enrichments with the GO biological processes, GO cellular components, and GO molecular functions databases screened out 12 candidate genes. In Step 3, 13 genes were found to be simultaneously up-regulated in the DEGs and a GEO dataset involving human diabetic retinal tissues. VEGFA and ERN1 were the 2 starred genes finally screened out by overlapping the 3 Steps. CONCLUSION: In this study, multiple genes were identified as crucial in the pathological process of RPE under protracted HG, providing potential candidates for future researches on DR. The current study highlights the importance of RPE in DR pathogenesis.

Identification and virus-induced gene silencing (VIGS) analysis of methyltransferase affecting tomato (Solanum lycopersicum) fruit ripening.

MAIN CONCLUSION: Six methyltransferase genes affecting tomato fruit ripening were identified through genome-wide screening, VIGS assay, and expression pattern analysis. The data provide the basis for understanding new mechanisms of methyltransferases. Fruit ripening is a critical stage for the formation of edible quality and seed maturation, which is finely modulated by kinds of factors, including genetic regulators, hormones, external signals, etc. Methyltransferases (MTases), important genetic regulators, play vital roles in plant development through epigenetic regulation, post-translational modification, or other mechanisms. However, the regulatory functions of numerous MTases except DNA methylation in fruit ripening remain limited so far. Here, six MTases, which act on different types of substrates, were identified to affect tomato fruit ripening. First, 35 MTase genes with relatively high expression at breaker (Br) stage of tomato fruit were screened from the tomato MTase gene database encompassing 421 genes totally. Thereafter, six MTase genes were identified as potential regulators of fruit ripening via virus-induced gene silencing (VIGS), including four genes with a positive regulatory role and two genes with a negative regulatory role, respectively. The expression of these six MTase genes exhibited diverse patterns during the fruit ripening process, and responded to various external ripening-related factors, including ethylene, 1-methylcyclopropene (1-MCP), temperature, and light exposure. These results help to further elaborate the biological mechanisms of MTase genes in tomato fruit ripening and enrich the understanding of the regulatory mechanisms of fruit ripening involving MTases, despite of DNA MTases.

The correlation study between the length and angle of ascending aortic and the incidence risk of acute type A aortic dissection.

Objective: This study utilized computed tomography angiography (CTA) to assess the risk of acute type A aortic dissection (ATAAD) by analyzing the imaging morphology indicators of the ascending aorta, along with the relevant risk factors associated with aortic dissection. Methods: The study utilized a retrospective observational research design. The population consisted of 172 patients who received treatment in the Department of Cardiothoracic Surgery at Qilu Hospital, Shandong University, from January 2018 to December 2022. The patients were divided into two groups: the ATAAD group (n = 97) and the thoracic aortic aneurysm group (TAA, n = 75). Demographic data and ascending aorta CTA measurements were collected from all patients. Single factor and multivariate logistic regression were employed to analyze the statistical differences in clinical data and ascending aorta CTA imaging morphology indicators between the two groups. Results: The variables were included in logistic multivariate analysis for further screening, indicating that the length of the ascending aorta (LAA) before ATAAD (OR = 3.365; 95% CI :1.742-6.500, P<0.001), ascending arch angle (asc-arch angle, OR = 0.902; 95% CI: 0.816-0.996, P = 0.042) and the maximum aortic diameter (MAD) before ATAAD, (OR = 0.614; 95% CI: 0.507-0.743, P<0.001) showed statistically significant differences. Conclusions: This study suggests that increased LAA and MAD, as well as a smaller asc-arch angle may be high-risk factors for the onset of ATAAD.

Comparison of condylar position after free fibular flap mandibular reconstruction using computer-assisted and traditional techniques.

OBJECTIVES: To compare the changes in condylar position after mandibular reconstruction with free fibular flap(FFF) and the differences between computer-assisted techniques and traditional methods on CT images. METHODS: Thirty-four patients who underwent mandibular reconstruction with free fibular flap were selected according to the inclusion and exclusion criteria. In the 3D group, virtual surgical planning (VSP) with osteotomy cutting plate and placement guiding plate were used, while the traditional group underwent freehand reconstruction. The CT data of 68 temporomandibular joints (TMJs) were recorded before and immediately after surgery. The condylar position was evaluated by measuring the anterior space (AS), posterior space (PS) and superior space (SS), and the ln (PS/AS) was calculated according to the method proposed by Pullinger and Hollender. RESULTS: In the patients included in the 3D group, the condyle on the ipsilateral side moved slightly backward; however, in the patients in the traditional group, the ipsilateral side moved considerably anteroinferior. No obvious changes on the contralateral side were noted. In the 3D group, 33% of ipsilateral condyles were in the posterior position postoperatively when compared with the preoperative position (13%). In the traditional group, the number of ipsilateral condyles in the anterior position increased from 4 to 10, accounting for 53% postoperatively. Contrary to the traditional group, the 3D group presented less condylar displacement on the ipsilateral side postoperatively. CONCLUSIONS: This study showed a decreased percentage of change in condylar position postoperatively when VSP was used. Virtual surgical planning improved the accuracy of FFF mandibular reconstruction and made the condylar position more stable.

Preparation of strong, UV-blocking and sustainable glucose-cross-linked cellulose plastics via hydroxyl-yne click reaction.

The construction of functional cellulose plastics possessing strong UV-blocking, hydrophilicity, and biodegradability is challenging. Therefore, we provide a novel strategy to successfully prepare sustainable and hydrophilic glucose-cross-linked cellulose (GC) plastics showing effective UV-blocking and excellent mechanical properties via hydroxyl-yne click reaction at room temperature. The results demonstrated that hydroxyl-yne click chemistry enabled efficient crosslinking of cellulose with glucose using 4-dimethylamino pyridine (DMAP) as a catalyst. Moreover, the DMAP residue imparted good UV-shielding properties to GC films exhibiting nearly 100 % UVC (200-275 nm) and 100 % UVB (320-275 nm) shielding ratios. The introduction of glucose imparted superior hydrophilicity (water contact angle of 40.3-43.2°) and improved water adsorption. Additionally, the mechanical properties of the GC films increased with the increasing crosslinking density, and the highest tensile stress was 94 MPa. The water-induced breaking and hydrogen bond reforming strategy led to a stress of 127 MPa and a strain of 25.6 % for the final GC2 film, which were excellent compared to those of the most reported cellulose films. Additionally, GC films were biosafe, exhibited improved oxygen barrier, and good biodegradability. Hence, this study provides a promising and efficient approach for preparing high-performance cellulose plastics.

Elastic Relaxor Ferroelectric by Thiol-ene Click Reaction.

As ferroelectrics hold significance and application prospects in wearable devices, the elastification of ferroelectrics becomes more and more important. Nevertheless, achieving elastic ferroelectrics requires stringent synthesis conditions, while the elastification of relaxor ferroelectric materials remains unexplored, presenting an untapped potential for utilization in energy storage and actuation for wearable electronics. The thiol-ene click reaction offers a mild and rapid reaction platform to prepare functional polymers. Therefore, we employed this approach to obtain an elastic relaxor ferroelectric by crosslinking an intramolecular carbon-carbon double bonds (CF=CH) polymer matrix with multiple thiol groups via a thiol-ene click reaction. The resulting elastic relaxor ferroelectric demonstrates pronounced relaxor-type ferroelectric behaviour. This material exhibits low modulus, excellent resilience, and fatigue resistance, maintaining a stable ferroelectric response even under strains up to 70 %. This study introduces a straightforward and efficient approach for the construction of elastic relaxor ferroelectrics, thereby expanding the application possibilities in wearable electronics.

Cecropin A Alleviates LPS-Induced Oxidative Stress and Apoptosis of Bovine Endometrial Epithelial Cells.

Dairy cows receiving a prolonged high-concentrate diet express an elevated concentration of lipopolysaccharides (LPSs) in the peripheral blood circulation, accompanied by a series of systemic inflammatory responses; however, the specific impacts of inflammation are yet to be determined. Cecropin-like antimicrobial peptides have become a research hotspot regarding antimicrobial peptides because of their excellent anti-inflammatory activities, and cecropin A is a major member of the cecropin family. To elucidate the mechanism of cecropin A as anti-inflammatory under the condition of sub-acute ruminal acidosis (SARA) in dairy cows, we induced inflammation in bEECs with LPS (10 µg/mL) and then added cecropin A (25 µM). Afterwards, we detected three categories of indexes including oxidative stress indices, inflammation-related genes, and apoptosis-related genes in bovine endometrial epithelial cells (bEECs). The results indicated that cecropin A has the ability to reduce inflammatory factors TNF-α, IL-1ß, and IL-8 and inhibit the MAPK pathway to alleviate inflammation. In addition, cecropin A is able to reduce reactive oxygen species (ROS) levels and alleviates LPS-induced oxidative stress and mitochondrial dysfunction by downregulating NADPH Oxidase (NOX), and upregulating catalase (CAT), glutathione peroxidase (GPX), and superoxide dismutase (SOD). Furthermore, cecropin A demonstrates the ability to inhibit apoptosis by suppressing the mitochondrial-dependent apoptotic pathway, specifically Fas/FasL-caspase-8/-3. The observed increase in the Bcl-2/Bax ratio, a known apoptosis regulator, further supports this finding. In conclusion, our study presents novel solutions for addressing inflammatory responses associated with SARA.

Oxygen-independent organic photosensitizer with ultralow-power NIR photoexcitation for tumor-specific photodynamic therapy.

Photodynamic therapy (PDT) is a promising cancer treatment but has limitations due to its dependence on oxygen and high-power-density photoexcitation. Here, we report polymer-based organic photosensitizers (PSs) through rational PS skeleton design and precise side-chain engineering to generate •O2- and •OH under oxygen-free conditions using ultralow-power 808 nm photoexcitation for tumor-specific photodynamic ablation. The designed organic PS skeletons can generate electron-hole pairs to sensitize H2O into •O2- and •OH under oxygen-free conditions with 808 nm photoexcitation, achieving NIR-photoexcited and oxygen-independent •O2- and •OH production. Further, compared with commonly used alkyl side chains, glycol oligomer as the PS side chain mitigates electron-hole recombination and offers more H2O molecules around the electron-hole pairs generated from the hydrophobic PS skeletons, which can yield 4-fold stronger •O2- and •OH production, thus allowing an ultralow-power photoexcitation to yield high PDT effect. Finally, the feasibility of developing activatable PSs for tumor-specific photodynamic therapy in female mice is further demonstrated under 808 nm irradiation with an ultralow-power of 15 mW cm-2. The study not only provides further insights into the PDT mechanism but also offers a general design guideline to develop an oxygen-independent organic PS using ultralow-power NIR photoexcitation for tumor-specific PDT.

WNT-inhibitory factor 1-mediated glycolysis protects photoreceptor cells in diabetic retinopathy.

BACKGROUND: In diabetic retinopathy (DR), hypoxia-inducible factor (HIF-1α) induces oxidative stress by upregulating glycolysis. This process leads to neurodegeneration, particularly photoreceptor cell damage, which further contributes to retinal microvascular deterioration. Further, the regulation of Wnt-inhibitory factor 1 (WIF1), a secreted Wnt signaling antagonist, has not been fully characterized in neurodegenerative eye diseases. We aimed to explore the impact of WIF1 on photoreceptor function within the context of DR. METHOD: Twelve-week-old C57BL/KsJ-db/db mice were intravitreally injected with WIF1 overexpression lentivirus. After 4 weeks, optical coherence tomography (OCT), transmission electron microscopy (TEM), H&E staining, and electroretinography (ERG) were used to assess the retinal tissue and function. The potential mechanism of action of WIF1 in photoreceptor cells was explored using single-cell RNA sequencing. Under high-glucose conditions, 661 W cells were used as an in vitro DR model. WIF1-mediated signaling pathway components were assessed using quantitative real-time PCR, immunostaining, and western blotting. RESULT: Typical diabetic manifestations were observed in db/db mice. Notably, the expression of WIF1 was decreased at the mRNA and protein levels. These pathological manifestations and visual function improved after WIF1 overexpression in db/db mice. TEM demonstrated that WIF1 restored damaged mitochondria, the Golgi apparatus, and photoreceptor outer segments. Moreover, ERG indicated the recovery of a-wave potential amplitude. Single-cell RNA sequencing and in vitro experiments suggested that WIF1 overexpression prevented the expression of glycolytic enzymes and lactate production by inhibiting the canonical Wnt signaling pathway, HIF-1α, and Glut1, thereby reducing retinal and cellular reactive oxygen species levels and maintaining 661 W cell viability. CONCLUSIONS: WIF1 exerts an inhibitory effect on the Wnt/ß-catenin-HIF-1α-Glut1 glycolytic pathway, thereby alleviating oxidative stress levels and mitigating pathological structural characteristics in retinal photoreceptor cells. This mechanism helps preserve the function of photoreceptor cells in DR and indicates that WIF1 holds promise as a potential therapeutic candidate for DR and other neurodegenerative ocular disorders.

Manganese drives ferroptosis of cancer cells via YAP/TAZ phase separation activated ACSL4 in OSCC.

OBJECTIVE: Ferroptosis has been defined as a novel form of regulated cell death characterized by iron-dependent lipid peroxidation. Manganese has been used to induce ferroptosis in cancer cells recently. This study aims to investigate whether manganese can induce ferroptosis in oral squamous cell carcinoma (OSCC) and the underlying biological mechanisms. MATERIALS AND METHODS: Cancer cells with or without manganese treatment were analyzed by RNA-sequencing to identify ferroptosis-related genes. Next, the activation of YAP/TAZ/ACSL4-ferroptosis signaling pathway was detected. Bioinformatic analysis and immunofluorescence assay were used to explore the phase separation of YAP/TAZ. Finally, specimens of OSCC patients were applied to analyze the clinical significance of YAP/TAZ/ACSL4. RESULTS: RNA-sequencing analysis showed the ferroptosis-related genes and YAP/TAZ were upregulated after manganese treatment. The results of immunofluorescence, ELISA, western blotting, etc. further confirmed that manganese-induced ferroptosis depends on YAP/TAZ/ACSL4 signaling pathway. Moreover, the activation of ACSL4 was achieved by YAP/TAZ phase separation. The survival analysis in OSCC specimen suggested that the higher level of YAP/TAZ-ACSL4 axis expression indicates longer survival. CONCLUSIONS: Manganese induces YAP/TAZ phase separation and subsequent ACSL4 activation via YAP/TAZ nuclear translocation, which facilitates ferroptosis of OSCC. Then YAP/TAZ-ACSL4 axis can be used as a potential prognostic predictor of OSCC patients.

Identification of particle distribution pattern in vertical profile via unmanned aerial vehicles observation.

Below the boundary layer, the air pollutants have been confirmed to present the decreasing trend with the height in most situaitons. However, the disperiosn rate of air pollutants in the vertical profile is rarely investigated in detail, especially through in-situ measurement. With this consideration, we employed an unmanned aerial vehicle equipped with portable monitoring equipments to scrutinize the vertical distribution of PM2.5. Based on the original data, we found that PM2.5 concentration decreases gradually with altitude below the boundary layer and demonstrated an obvious linear correlation. Therefore, the vertical distribution of PM2.5 was quantified by representing the distribution of PM2.5 with the slope of PM2.5 vertical distribution. We used backward trajectories to reveal the causes of outliers (PM2.5 increasing with altitude), and found that PM2.5 in the high altitude came from the southwest. Besides, the relationship between the vertical distribution of PM2.5 and various meteorological factors was investigated using stepwise regression analysis. The results show that the four meteorological factors most strongly correlated with the slope values are: (a) the difference in relative humidity between the ground and the air; (b) the difference in temperature between the ground and the air; (c) the height of the boundary layer; and (d) the wind speed. The slope values increase with increasing the difference in relative humidity between ground and air and the difference in temperature between the ground and the air, and decrease with increasing boundary layer height and wind speed. According to the Random Forest calculations, the ground-to-air relative humidity difference is the most important at 0.718; the wind speed is the least important at 0.053; and the ground-to-air temperature difference and boundary layer height are 0.140 and 0.088, respectively.

Cardiac tissue model of immune-induced dysfunction reveals the role of free mitochondrial DNA and the therapeutic effects of exosomes.

Despite tremendous progress in the development of mature heart-on-a-chip models, human cell-based models of myocardial inflammation are lacking. Here, we bioengineered a vascularized heart-on-a-chip with circulating immune cells to model severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-induced acute myocarditis. We observed hallmarks of coronavirus disease (COVID-19)-induced myocardial inflammation, as the presence of immune cells augmented the secretion of proinflammatory cytokines, triggered progressive impairment of contractile function, and altered intracellular calcium transients. An elevation of circulating cell-free mitochondrial DNA (ccf-mtDNA) was measured first in the heart-on-a-chip and then validated in COVID-19 patients with low left ventricular ejection fraction, demonstrating that mitochondrial damage is an important pathophysiological hallmark of inflammation-induced cardiac dysfunction. Leveraging this platform in the context of SARS-CoV-2-induced myocardial inflammation, we established that administration of endothelial cell-derived exosomes effectively rescued the contractile deficit, normalized calcium handling, elevated the contraction force, and reduced the ccf-mtDNA and cytokine release via Toll-like receptor-nuclear factor κB signaling axis.

Comparison on emulsifying and emulgelling properties of low methoxyl pectin with varied degree of methoxylation from different de-esterification methods.

Low methoxyl pectin (LMP) with different degree of methoxylation (DM, 40-50 %, 20-30 % and 5-10 %) were prepared from commercially available citrus pectin using high hydrostatic pressure assisted enzymatic (HHP-pectin) and traditional alkaline (A-pectin) de-esterification method. The results showed that both de-esterification methods and DM exhibited LMPs with varied physicochemical, structural, and functional properties. As the DM decreased, LMP showed a decrease in molecular weight (Mw), while an increase in negative charges and rhamnogalacturonan I (RG-I) ratio, accompanied with better emulsion stability, emulsion gel strength and water-holding properties. Relative to A-pectin, HHP-pectin had higher Mw and lower RG-I side chain ratio, contributing to its better thermal stability, apparent viscosity, and emulgelling properties. HHP-pectin with lower DM (5-10 %) showed superior thickening, emulsifying and emulgelling properties, while that with higher DM (40-45 %) had superior thermal stability, which provided alternative for de-esterification and targeted structural modification of pectin.

The evolution law of deviatoric stress and asymmetric control technology in roadways during panel mining through overlying residual coal pillars.

Close-distance coal seams (CDCS) are widely distributed, and the layout of the upper and lower panels can be divided into "=" type and "+" type. The "+" superposition of upper and lower coal pillars in CDCS caused strong mine pressure, but there are few studies on the panel crossing residual coal pillars (RCP) when the upper and lower coal seams are "+" type layout. In view of the special spatial position ("+" type layout), this paper takes the typical panel 4-301 of a particular mine as the project indagation background and studies mining and crossing the overlying coal pillars by dint of field measurement, numerical simulation, indoor test, and engineering application. Compared with vertical stress or horizontal stress alone, the indexes of deviatoric stress and plastic zone can reflect the failure evolution of surrounding rock more comprehensively. Hence, this paper analyzes the expansion form of the plastic zone and the variation law of deviatoric stress before and after mining influence in the underlying mining roadway. The research results show that: (1) There is a sub-peak zone of deviatoric stress under the RCP. The deviatoric stress is bimodal in the range of 9 m below. After the peak value decays to 7.4 MPa, it changes to a single peak located in the area directly below the middle of the RCP. (2) The maximum plastic zones of the roof and two ribs of the roadway below the RCP are 3.4 m and 5 m, respectively. The crest value of deviatoric stress reaches 10 MPa. As the distance between the panel and the RCP decreases, the shape of the high deviatoric stress area presents the evolution law from the "ellipse" of the roof → the "crescent" of two ribs → the "cochlea" of the tips of the ribs. (3) When the mining of the underlying panel is 10 m, 0 m, or - 10 m away from the RCP (without passing through the RCP). The crest value of deviatoric stress within 5-10 m in advance of the roadway increases in turn. However, the peak value is significantly reduced when it is - 20 m away from the RCP (through the RCP). The crest value of deviatoric stress of two ribs decreases in turn along the panel rib → section coal pillar rib → solid coal rib. Based on this, the underlying 45 m of the RCP is divided into area I (10 m), area II (overlapping area 20 m), and area III (15 m) based on the degree of disturbance. And propose the technical scheme of asymmetric combined control in different zones by using asymmetric channel steel truss anchor cable for the top-ribs of areas I and III, and top-ribs asymmetric channel steel truss anchor cable + door-type support in area II. On-site project practice shows that the partitioned control technology successfully resisted the roadway instability and failure caused by the dynamic-static superimposed stress disturbance under the RCP and realized the primary support of the sectional coal roadway. The conclusion provides technical support and scheme design for the partitioning support of roadways under similar "+" type cross-panels.

Cooperation between autotrophic and heterotrophic denitrifiers under low C/N ratios revealed by individual-based modelling.

Denitrifying biofilms, in which autotrophic denitrifiers (AD) and heterotrophic denitrifiers (HD) coexist, play a crucial role in removing nitrate from water or wastewater. However, it is difficult to elucidate the interactions between HD and AD through sequencing-based experimental methods. Here, we developed an individual-based model to describe the interspecies dynamics and priority effects between sulfur-based AD (Thiobacillus denitrificans) and HD (Thauera phenylcarboxya) under different C/N ratios. In test I (coexistence simulation), AD and HD were initially inoculated at a ratio of 1:1. The simulation results showed excellent denitrification performance and a coaggregation pattern of denitrifiers, indicating that cooperation was the predominant interaction at a C/N ratio of 0.25 to 1.5. In test II (invasion simulation), in which only one type of denitrifier was initially inoculated and the other was added at the invasion time, denitrifiers exhibited a stratification pattern in biofilms. When HD invaded AD, the final HD abundance decreased with increasing invasion time, indicating an enhanced priority effect. When AD invaded HD, insufficient organic carbon sources weakened the priority effect by limiting the growth of HD populations. This study reveals the interaction between autotrophic and heterotrophic denitrifiers, providing guidance for optimizing wastewater treatment process.

FARS2 Deficiency Causes Cardiomyopathy by Disrupting Mitochondrial Homeostasis and the Mitochondrial Quality Control System.

BACKGROUND: Hypertrophic cardiomyopathy (HCM) is a common heritable heart disease. Although HCM has been reported to be associated with many variants of genes involved in sarcomeric protein biomechanics, pathogenic genes have not been identified in patients with partial HCM. FARS2 (the mitochondrial phenylalanyl-tRNA synthetase), a type of mitochondrial aminoacyl-tRNA synthetase, plays a role in the mitochondrial translation machinery. Several variants of FARS2 have been suggested to cause neurological disorders; however, FARS2-associated diseases involving other organs have not been reported. We identified FARS2 as a potential novel pathogenic gene in cardiomyopathy and investigated its effects on mitochondrial homeostasis and the cardiomyopathy phenotype. METHODS: FARS2 variants in patients with HCM were identified using whole-exome sequencing, Sanger sequencing, molecular docking analyses, and cell model investigation. Fars2 conditional mutant (p.R415L) or knockout mice, fars2-knockdown zebrafish, and Fars2-knockdown neonatal rat ventricular myocytes were engineered to construct FARS2 deficiency models both in vivo and in vitro. The effects of FARS2 and its role in mitochondrial homeostasis were subsequently evaluated using RNA sequencing and mitochondrial functional analyses. Myocardial tissues from patients were used for further verification. RESULTS: We identified 7 unreported FARS2 variants in patients with HCM. Heart-specific Fars2-deficient mice presented cardiac hypertrophy, left ventricular dilation, progressive heart failure accompanied by myocardial and mitochondrial dysfunction, and a short life span. Heterozygous cardiac-specific Fars2R415L mice displayed a tendency to cardiac hypertrophy at age 4 weeks, accompanied by myocardial dysfunction. In addition, fars2-knockdown zebrafish presented pericardial edema and heart failure. FARS2 deficiency impaired mitochondrial homeostasis by directly blocking the aminoacylation of mt-tRNAPhe and inhibiting the synthesis of mitochondrial proteins, ultimately contributing to an imbalanced mitochondrial quality control system by accelerating mitochondrial hyperfragmentation and disrupting mitochondrion-related autophagy. Interfering with the mitochondrial quality control system using adeno-associated virus 9 or specific inhibitors mitigated the cardiac and mitochondrial dysfunction triggered by FARS2 deficiency by restoring mitochondrial homeostasis. CONCLUSIONS: Our findings unveil the previously unrecognized role of FARS2 in heart and mitochondrial homeostasis. This study may provide new insights into the molecular diagnosis and prevention of heritable cardiomyopathy as well as therapeutic options for FARS2-associated cardiomyopathy.

Supramolecular Genome Editing: Targeted Delivery and Endogenous Activation of CRISPR/Cas9 by Dynamic Host-Guest Recognition.

We synthesize supramolecular poly(disulfide) (CPS) containing covalently attached cucurbit[7]uril (CB[7]), which is exploited not only as a carrier to deliver plasmid DNA encoding destabilized Cas9 (dsCas9), but also as a host to include trimethoprim (TMP) by CB[7] moieties through the supramolecular complexation to form TMP@CPS/dsCas9. Once the plasmid is transfected into tumor cells by CPS, the presence of polyamines can competitively trigger the decomplexation of TMP@CPS, thereby displacing and releasing TMP from CB[7] to stabilize dsCas9 that can target and edit the genomic locus of PLK1 to inhibit the growth of tumor cells. Following the systemic administration of TMP@CPS/dsCas9 decorated with hyaluronic acid (HA), tumor-specific editing of PLK1 is detected due to the elevated polyamines in tumor microenvironment, greatly minimizing off-target editing in healthy tissues and non-targeted organs. As the metabolism of polyamines is dysregulated in a wide range of disorders, this study offers a supramolecular approach to precisely control CRISPR/Cas9 functions under particular pathological contexts.

Regulating Lipid Metabolism via Mitochondrial Dynamics in Tongue Squamous Cell Carcinoma Cancer Stem Cells.

BACKGROUND: Cancer stem cells (CSCs) are a sub-population of cancer cells present in many kinds of malignant tumors that have the potential for self-proliferation and differentiation. These cells have been demonstrated as the main cause of tumor recurrence and metastasis. Strong evidence indicates that CSCs prefer reprogrammed fatty acid ß-oxidation over oxidative phosphorylation for sustaining energy supply. Although mitochondrial dynamics participate in the regulation of cancer stemness, the correlation between the inhibition of mitochondrial fission and the regulation of lipid metabolism in CSCs remains poorly understood. METHODS: The human tongue squamous cell carcinoma (TSCC) cell lines CAL27 and SAS were used to obtain the CSCs by 3D Spheroid Culture. Then,western blot methods, RT-PCR and flow cytometry analysis were used to identify the TSCC CSCs. Next, Immunofluorescence method, transmission electron microscopy detection and western blot methods were used to evaluate the mitochondrial morphology and the quantity of lipid droplets (LDs). Lastly, lipidomic analysis was applied to explored the lipidomic alterations of TSCC CSCs with different mitochondrial morphology. RESULTS: Here, we show that the quantity of lipid droplets containing intracellular triglyceride (TG) can be decreased by regulating mitochondrial morphology. Lipidomic analysis using ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) also compared alterations in lipid metabolites in tongue squamous cell carcinoma (TSCC) CSCs, TSCC cells (non-CSCs), and CSCs with different mitochondrial morphology. Discriminant lipids of statistical significance were successfully annotated, including phosphatidylcholines (PCs), phosphatidylethanolamines (PEs), sphingomyelins (SMs), triacylglycerols (TGs), phosphatidylglycerols (PGs), phosphatidylserines (PSs), lysophosphatidylcholines (LPCs), and lysophosphatidylethanolamines (LPEs). CONCLUSION: This study provides a deeper insight into the alterations of lipid metabolism associated with TSCC CSCs, non-CSCs and CSCs regulated by mitochondrial dynamics and thus serves as a guide toward novel targeted therapies.