In Vivo Base Editing of Scn5a Rescues Type 3 Long QT Syndrome in Mice.

BACKGROUND: Pathogenic variants in SCN5A can result in long QT syndrome type 3, a life-threatening genetic disease. Adenine base editors can convert targeted A T base pairs to G C base pairs, offering a promising tool to correct pathogenic variants. METHODS: We generated a long QT syndrome type 3 mouse model by introducing the T1307M pathogenic variant into the Scn5a gene. The adenine base editor was split into 2 smaller parts and delivered into the heart by adeno-associated virus serotype 9 (AAV9-ABEmax) to correct the T1307M pathogenic variant. RESULTS: Both homozygous and heterozygous T1307M mice showed significant QT prolongation. Carbachol administration induced Torsades de Pointes or ventricular tachycardia for homozygous T1307M mice (20%) but not for heterozygous or wild-type mice. A single intraperitoneal injection of AAV9-ABEmax at postnatal day 14 resulted in up to 99.20% Scn5a transcripts corrected in T1307M mice. Scn5a mRNA correction rate >60% eliminated QT prolongation; Scn5a mRNA correction rate <60% alleviated QT prolongation. Partial Scn5a correction resulted in cardiomyocytes heterogeneity, which did not induce severe arrhythmias. We did not detect off-target DNA or RNA editing events in ABEmax-treated mouse hearts. CONCLUSIONS: These findings show that in vivo AAV9-ABEmax editing can correct the variant Scn5a allele, effectively ameliorating arrhythmia phenotypes. Our results offer a proof of concept for the treatment of hereditary arrhythmias.

Histone demethylase KDM1A promotes hepatic steatosis and inflammation by increasing chromatin accessibility in NAFLD.

Nonalcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease without specific Food and Drug Administration-approved drugs. Recent advances suggest that chromatin remodeling and epigenetic alteration contribute to the development of NAFLD. The functions of the corresponding molecular modulator in NAFLD, however, are still elusive. KDM1A, commonly known as lysine-specific histone demethylase 1, has been reported to increase glucose uptake in hepatocellular carcinoma. In addition, a recent study suggests that inhibition of KDM1A reduces lipid accumulation in primary brown adipocytes. We here investigated the role of KDM1A, one of the most important histone demethylases, in NAFLD. In this study, we observed a significant upregulation of KDM1A in NAFLD mice, monkeys, and humans compared to the control group. Based on these results, we further found that the KDM1A can exacerbate lipid accumulation and inflammation in hepatocytes and mice. Mechanistically, KDM1A exerted its effects by elevating chromatin accessibility, subsequently promoting the development of NAFLD. Furthermore, the mutation of KDM1A blunted its capability to promote the development of NAFLD. In summary, our study discovered that KDM1A exacerbates hepatic steatosis and inflammation in NAFLD via increasing chromatin accessibility, further indicating the importance of harnessing chromatin remodeling and epigenetic alteration in combating NAFLD. KDM1A might be considered as a potential therapeutic target in this regard.

The impact of Karnofsky performance status on prognosis of patients with hepatocellular carcinoma in liver transplantation.

BACKGROUND: Functional performance as measured by the Karnofsky Performance Status (KPS) scale has been linked to the outcomes of liver transplant patients; however, the effect of KPS on the outcomes of the hepatocellular carcinoma (HCC) liver transplant population has not been fully elucidated. We aimed to investigate the association between pre-transplant KPS score and long-term outcomes in HCC patients listed for liver transplantation. METHODS: Adult HCC candidates listed on the Scientific Registry of Transplant Recipients (SRTR) database from January 1, 2011 to December 31, 2017 were grouped into group I (KPS 80-100%, n = 8,379), group II (KPS 50-70%, n = 8,091), and group III (KPS 10-40%, n = 1,256) based on percentage KPS score at listing. Survival was compared and multivariable analysis was performed to identify independent predictors. RESULTS: Patients with low KPS score had a higher risk of removal from the waiting list. The 5-year intent-to-treat survival was 57.7% in group I, 53.2% in group II and 46.7% in group III (P < 0.001). The corresponding overall survival was 77.6%, 73.7% and 66.3% in three groups, respectively (P < 0.001). Multivariable analysis demonstrated that KPS was an independent predictor of intent-to-treat survival (P < 0.001, reference group I; HR 1.19 [95%CI 1.07-1.31] for group II, P = 0.001; HR 1.63 [95%CI 1.34-1.99] for group III, P < 0.001) and overall survival(P < 0.001, reference group I; HR 1.16 [95%CI 1.05-1.28] for group II, P = 0.004; HR 1.53 [95%CI 1.26-1.87] for group III, P < 0.001). The cumulative 5-year recurrence rates was higher in group III patients (7.4%), compared with 5.2% in group I and 5.5% in group II (P = 0.037). However, this was not significant in the competing regression analysis. CONCLUSIONS: Low pre-transplant KPS score is associated with inferior long-term survival in liver transplant HCC patients, but is not significantly associated with post-transplant tumor recurrence.

Point Cloud Registration Method Based on Geometric Constraint and Transformation Evaluation.

Existing point-to-point registration methods often suffer from inaccuracies caused by erroneous matches and noisy correspondences, leading to significant decreases in registration accuracy and efficiency. To address these challenges, this paper presents a new coarse registration method based on a geometric constraint and a matrix evaluation. Compared to traditional registration methods that require a minimum of three correspondences to complete the registration, the proposed method only requires two correspondences to generate a transformation matrix. Additionally, by using geometric constraints to select out high-quality correspondences and evaluating the matrix, we greatly increase the likelihood of finding the optimal result. In the proposed method, we first employ a combination of descriptors and keypoint detection techniques to generate initial correspondences. Next, we utilize the nearest neighbor similarity ratio (NNSR) to select high-quality correspondences. Subsequently, we evaluate the quality of these correspondences using rigidity constraints and salient points' distance constraints, favoring higher-scoring correspondences. For each selected correspondence pair, we compute the rotation and translation matrix based on their centroids and local reference frames. With the transformation matrices of the source and target point clouds known, we deduce the transformation matrix of the source point cloud in reverse. To identify the best-transformed point cloud, we propose an evaluation method based on the overlap ratio and inliers points. Through parameter experiments, we investigate the performance of the proposed method under various parameter settings. By conducting comparative experiments, we verified that the proposed method's geometric constraints, evaluation methods, and transformation matrix computation consistently outperformed other methods in terms of root mean square error (RMSE) values. Additionally, we validated that our chosen combination for generating initial correspondences outperforms other descriptor and keypoint detection combinations in terms of the registration result accuracy. Furthermore, we compared our method with several feature-matching registration methods, and the results demonstrate the superior accuracy of our approach. Ultimately, by testing the proposed method on various types of point cloud datasets, we convincingly established its effectiveness. Based on the evaluation and selection of correspondences and the registration result's quality, our proposed method offers a solution with fewer iterations and higher accuracy.

Recurrence and malignant transformation of laryngeal leukoplakia treated with CO2 laser: A systematic review and meta-analysis.

BACKGROUND: Laryngeal leukoplakia (LL) is a white lesion with high potential of recurrence and malignant transformation. Currently, CO2 laser has become the primary surgical treatment for LL, and the recurrence and malignant transformation rates after treatment vary widely. OBJECTIVE: We performed a systematic review and meta-analysis dedicated to evaluating the rates of recurrence and malignant transformation of LL lesions treated with CO2 laser and exploring relevant risk factors for recurrence or malignant transformation. METHODS: Literature searches were conducted on ProQuest, PubMed, Web of Science, Ovid Medline, Embase, and Cochrane databases. Some articles identified through hand searching were included. RESULTS: A total of 14 articles and 1462 patients were included in this review. Pooled results showed that the overall recurrence rate was 15%, and the malignant transformation rate was 3%. Subgroup analysis showed that the dysplasia grade was not a significant risk factor for the recurrence and malignant transformation of LL (P > .05). CONCLUSIONS: The results of this systematic review and meta-analysis suggest that the CO2 laser is a safe and effective surgical instrument for the excision of LL, which yields low rates of recurrence and malignant transformation. The risk factors relevant to recurrence or malignant transformation remain unclear and require further investigation.

Plasmon-enhanced deep ultraviolet Micro-LED arrays for solar-blind communications.

Localized surface plasmon resonance (LSPR)-enhanced deep ultraviolet (DUV) Micro-light emitting diodes (Micro-LEDs) using Al nanotriangle arrays (NTAs) are reported for improving the -3 dB modulation bandwidth. Through self-assembled nanospheres, the high-density Al NTAs arrays are transferred into the designated p-AlGaN region of the Micro-LEDs, realizing the effect of LSPR coupling. A 2.5-fold enhancement in photoluminescence (PL) intensity is demonstrated. Combined with the PL intensity ratio at 300 K and 10 K, internal quantum efficiency (IQE) may be increased about 15-20% by the plasmonic effect and the carrier lifetime decreases from 1.15 ns to 0.82 ns, suggesting that LSPR accelerates the spontaneous emission rate. Resulting from the improvement of the IQE, the electroluminescence intensity of Micro-LED arrays with LSPR is obviously increased. Meanwhile, the -3 dB bandwidth of 6 × 6 Micro-LED arrays is increased from 180 MHz to 300 MHz at a current density of 200 A/cm2. A potential way is proposed to further increase both the IQE and the modulation bandwidth of DUV Micro-LEDs.

Efficient Correction of a Hypertrophic Cardiomyopathy Mutation by ABEmax-NG.

[Figure: see text].

Method to Solve Underwater Laser Weak Waves and Superimposed Waves.

With the rapid development of Lidar technology, the use of Lidar for underwater terrain detection has become feasible. There is still a challenge in the process of signal resolution: the underwater laser echo signal is different to propagating in the air, and it is easy to produce weak waves and superimposed waves. However, existing waveform decomposition methods are not effective in processing these waveform signals, and the underwater waveform signal cannot be correctly decomposed, resulting in subsequent data-processing errors. To address these issues, this study used a drone equipped with a 532 nm laser to detect a pond as the study background. This paper proposes an improved inflection point selection decomposition method to estimate the parameter. By comparing it with other decomposition methods, we found that the RMSE is 2.544 and R2 is 0.995975, which is more stable and accurate. After estimating the parameters, this study used oscillating particle swarm optimization (OPSO) and the Levenberg-Marquardt algorithm (LM) to optimize the estimated parameters; the final results show that the method in this paper is closer to the original waveform. In order to verify the processing effect of the method on complex waveform, this paper decomposes and optimizes the simulated complex waveforms; the final RMSE is 0.0016, R2 is 1, and the Gaussian component after decomposition can fully represent the original waveform. This method is better than other decomposition methods in complex waveform decomposition, especially regarding weak waves and superimposed waves.

[Clinical and genetic characteristics of 12 cases of Loeys-Dietz syndrome].

OBJECTIVE: To summarize the clinical features and spectrum of genetic variants in 12 patients with Loeys-Dietz syndrome (LDS), and to explore the correlation between the type of genetic variants and clinical phenotypes. METHODS: Twelve patients suspected for LDS at Beijing Anzhen Hospital Affiliated to Capital Medical University from January 2015 to January 2022 were selected as the study subjects. Clinical data of the patients were collected. Genomic DNA was extracted from peripheral blood samples and subjected to genetic testing. Pathogenicity of candidate variants was analyzed. RESULTS: The clinical phenotypes of the 12 patients have mainly included cardiovascular, musculoskeletal, craniofacial, skin, ocular and other systemic signs. Four patients (patients 5-1, 5-2, 6, 7) have carried heterozygous missense variants of the TGFBR1 gene, 5 patients (patients 1-1, 1-2, 2, 3, 4) have carried heterozygous variants of the TGFBR2 gene, and 2 patients (patients 8-1, 8-2) had carried heterozygous frameshift variants of the TGFB3 gene. One patient (patient 9) had carried a heterozygous missense variant of the SMAD3 gene. Among these, TGFBR1 c.603T>G (p.1201M) and TGFB3 c.536delA (p.H179FS35) had not been reported previously. CONCLUSION: Variants of the TGFBR1, TGFBR2, SMAD3, TGFB2, TGFB3 and SMAD2 genes are mainly associated with LDS. The severity of the disease phenotype caused by the same variant may vary, whilst the clinical phenotype caused by different variant sites may be specific.

GaN-based parallel micro-light-emitting diode arrays with dual-wavelength InxGa1-xN/GaN MQWs for visible light communication.

The dual-wavelength InxGa1-xN/GaN micro light emitting diode (Micro-LED) arrays are fabricated by flip-chip parallel connection. It is noted that the Micro-LED arrays with smaller diameter present considerably bigger light output power density (LOPD). For all Micro-LEDs, the LOPD increases continuously with increasing injection current density until it "turns over". It also can be observed that the maximum value of LOPD is determined by the blue quantum well (QW) for the broad area LED. In comparison, the green peak intensity dominates the change of LOPD in the Micro-LEDs. In addition, the enhancement of the green peak intensity value for the Micro-LEDs are considered as a consequence of the combined effects of the reduction in the quantum-confined Stark effect (QCSE) and the crowding effect, high LEE as well as geometric shape. Moreover, -3dB modulation bandwidths of the four different kinds of Micro-LEDs increase with the decrease of the device diameter in the same injected current density, higher than that of the broad area LED. The -3dB modulation bandwidth of the 60 µm Micro-LED shows 1.4 times enhancement compared to that of the broad area LED under the current density of 300 mA/cm2. Evidently, the dual-wavelength InxGa1-xN/GaN Micro-LEDs have great potential in both solid-state lighting (SSL) and the visible light communication (VLC) in the future fabrication.

Determination of meloxicam in human plasma by ultra-high-performance liquid chromatography-tandem mass spectrometry and its application in a pharmacokinetic study.

A rapid, selective and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to detect meloxicam in human plasma. A triple quadrupole tandem mass spectrometer equipped with an electrospray ionization source was used in positive ion mode. Protein precipitation with acetonitrile was used for sample preparation. Meloxicam and 13 C6 -meloxicam internal standard were analyzed on an Acquity CSH C18 column with a mobile phase of acetonitrile and water in 0.1% formic acid using a gradient program for separation. The retention time of meloxicam was 1.1 min and the total run time was only 2.0 min. Detection was performed in multiple reaction monitoring mode using an electrospray ionization source with optimized mass spectrometry parameters. The calibration curves were linear in the range 10.0-3.00 × 103 ng/ml (r ≥ 0.99). The within-run and between-run RSDs were ≤14.8%. The within-run and between-run REs ranged from -4.6 to 10.7%. There was no significant matrix effect, and the recovery rate was high. This method was fully validated, including reinjection reproducibility in human plasma. The method was applied to the pharmacokinetic study. All of the incurred sample reanalysis methods met the criteria.

Knockout of MYOM1 in human cardiomyocytes leads to myocardial atrophy via impairing calcium homeostasis.

Myomesin-1 (encoded by MYOM1 gene) is expressed in almost all cross-striated muscles, whose family (together with myomesin-2 and myomesin-3) helps to cross-link adjacent myosin to form the M-line in myofibrils. However, little is known about its biological function, causal relationship and mechanisms underlying the MYOM1-related myopathies (especially in the heart). Regrettably, there is no MYMO1 knockout model for its study so far. A better and further understanding of MYOM1 biology is urgently needed. Here, we used CRISPR/Cas9 gene-editing technology to establish an MYOM1 knockout human embryonic stem cell line (MYOM1-/- hESC), which was then differentiated into myomesin-1 deficient cardiomyocytes (MYOM1-/- hESC-CMs) in vitro. We found that myomesin-1 plays an important role in sarcomere assembly, contractility regulation and cardiomyocytes development. Moreover, myomesin-1-deficient hESC-CMs can recapitulate myocardial atrophy phenotype in vitro. Based on this model, not only the biological function of MYOM1, but also the aetiology, pathogenesis, and potential treatments of myocardial atrophy caused by myomesin-1 deficiency can be studied.

Microscale grooves regulate maturation development of hPSC-CMs by the transient receptor potential channels (TRP channels).

The use of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) is limited in drug discovery and cardiac disease mechanism studies due to cell immaturity. Micro-scaled grooves can promote the maturation of cardiomyocytes by aligning them in order, but the mechanism of cardiomyocytes alignment has not been studied. From the level of calcium activity, gene expression and cell morphology, we verified that the W20H5 grooves can effectively promote the maturation of cardiomyocytes. The transient receptor potential channels (TRP channels) also play an important role in the maturation and development of cardiomyocytes. These findings support the engineered hPSC-CMs as a powerful model to study cardiac disease mechanism and partly mimic the myocardial morphological development. The important role of the TRP channels in the maturation and development of myocardium is first revealed.

Integrative metabolic profile of myelodysplastic syndrome based on UHPLC-MS.

Myelodysplastic syndrome (MDS) is a neoplastic disease originating from hematopoietic stem cells. Currently, hematopoietic stem cell transplantation (HSCT) is the most effective cure, although lenalidomide, azacytidine, and decitabine have been applied to relieve symptoms of MDS. The purpose of this study was to evaluate the changes in endogenous metabolites by applying a UHPLC-MS (ultra-high-performance liquid chromatography-MS) metabolomics approach and to investigate metabolic pathways related to MDS. An untargeted metabolomics approach based on UHPLC-MS in combination with multivariate data analysis, including partial least squares discrimination analysis and orthogonal partial least squares discriminant analysis, was established to investigate potential biomarkers in the plasma of MDS patients. As a result, 29 biomarkers were identified to distinguish between MDS patients, HSCT patients, and healthy controls, which were mainly related to inflammation regulation, amino acid metabolism, fatty acid metabolism, and energy metabolism. To our knowledge, this is the first time where plasma metabolomics was combined with HSCT to study the pathogenesis and therapeutic target of MDS. The identification of biomarkers and analysis of metabolic pathways could offer the possibility of discovering new therapeutic targets for MDS in the future.

Optimization-Based Approaches for Minimizing Deployment Costs for Wireless Sensor Networks with Bounded Estimation Errors.

As wireless sensor networks have become more prevalent, data from sensors in daily life are constantly being recorded. Due to cost or energy consumption considerations, optimization-based approaches are proposed to reduce deployed sensors and yield results within the error tolerance. The correlation-aware method is also designed in a mathematical model that combines theoretical and practical perspectives. The sensor deployment strategies, including XGBoost, Pearson correlation, and Lagrangian Relaxation (LR), are determined to minimize deployment costs while maintaining estimation errors below a given threshold. Moreover, the results significantly ensure the accuracy of the gathered information while minimizing the cost of deployment and maximizing the lifetime of the WSN. Furthermore, the proposed solution can be readily applied to sensor distribution problems in various fields.

"Optical tentacle" of suspended polymer micro-rings on a multicore fiber facet for vapor sensing.

We designed a new type of gas sensor, an optical tentacle, made of highly integrated polymer micro-ring resonators in three-dimensional space on the tiny end-facet of a multicore optical fiber. Two pairs of three polymer micro-ring resonators were hung symmetrically on both sides of three suspended micro-waveguides as the sensing units. The micro-waveguides interlace to form a three-layer nested configuration, which makes the multicore optical fiber a "tentacle" for vapors of volatile organic compounds. Both experiments and theoretical simulation confirmed that the symmetrical coupling of multiple pairs of rings with the micro-waveguide had better resonance than the single ring setup. This is because the symmetrical light modes in the waveguides couple with the rings separately. All the optical micro-components were fabricated by the two-photon lithography technology on the end facet of multicore optical fiber. The optical tentacle shows good sensitivity and reversibility. This approach can also be adopted for sensor array design on a chip. Furthermore, optical sensors that can sense vapors with multiple constituents may be achieved in the future by adding selective sensitive materials to or on the surface of the rings.

Screening of BHK-21 cellular proteins that interact with outer membrane protein 43K OMP of Fusobacterium necrophorum.

Fusobacterium necrophorum is a Gram negative, spore-free, anaerobic bacterium that can cause pyogenic and necrotic infections in animals and humans. It is a major bovine pathogen and causes hepatic abscesses, foot rot, and necrotic laryngitis. The 43K OMP of F. necrophorum is an outer membrane protein with molecular weight of 43 kDa, exhibiting similarity to pore-forming proteins of other Fusobacterium species that plays an important role in bacterial infections. However, the role of 43K OMP in F. necrophorum adhesion remains unknown. In this study, we evaluated whether the 43K OMP of F. necrophorum mediates adhesion to BHK-21 cells and performed a preliminary screen of the proteins that interact with 43K OMP of F. necrophorum by immunoprecipitation-mass spectrometry. The results showed that the natural 43K OMP and recombinant 43K OMP could bind to BHK-21 cells, and preincubation of F. necrophorum with an antibody against the recombinant 43K OMP of F. necrophorum decreased binding to BHK-21 cells. Seventy differential interacting proteins were successfully screened by immunoprecipitation-mass spectrometry. Among these seventy differential interacting proteins, seven cell membrane proteins and four extracellular matrix proteins shown to be relevant to bacteria adhesion through subcellular localization and single-molecule function analysis. These data increase our understanding of the pathogenesis of F. necrophorum and provide a new theoretical basis for the design of antimicrobial drugs against F. necrophorum.

Long-term dynamics of mycorrhizal root tips in a loblolly pine forest grown with free-air CO2 enrichment and soil N fertilization for 6 years.

Large-scale, long-term FACE (Free-Air CO2 enrichment) experiments indicate that increases in atmospheric CO2 concentrations will influence forest C cycling in unpredictable ways. It has been recently suggested that responses of mycorrhizal fungi could determine whether forest net primary productivity (NPP) is increased by elevated CO2 over long time periods and if forests soils will function as sources or sinks of C in the future. We studied the dynamic responses of ectomycorrhizae to N fertilization and atmospheric CO2 enrichment at the Duke FACE experiment using minirhizotrons over a 6 year period (2005-2010). Stimulation of mycorrhizal production by elevated CO2 was observed during only 1 (2007) of 6 years. This increased the standing crop of mycorrhizal tips during 2007 and 2008; during 2008, significantly higher mortality returned standing crop to ambient levels for the remainder of the experiment. It is therefore unlikely that increased production of mycorrhizal tips can explain the lack of progressive nitrogen limitations and associated increases in N uptake observed in CO2 -enriched plots at this site. Fertilization generally decreased tree reliance on mycorrhizae as tip production declined with the addition of nitrogen as has been shown in many other studies. Annual NPP of mycorrhizal tips was greatest during years with warm January temperatures and during years with cool spring temperatures. A 2 °C increase in average late spring temperatures (May and June) decreased annual production of mycorrhizal root tip length by 50%. This has important implications for ecosystem function in a warmer world in addition to potential for forest soils to sequester atmospheric C.

Cysteine and thiosulfate promoted cadmium immobilization in strain G303 by the formation of extracellular CdS.

Bacteria have evolved a variety of strategies to defend themselves against cadmium toxicity, however, the specific mechanisms involved in the enhancement of bacterial cadmium resistance by sulfur sources are unclear. In this study, a novel cadmium (Cd)-tolerant bacterium, Stenotrophomonas geniculata G303, was isolated from activated sludge. The growth of strain G303 under diverse Cd concentrations was investigated, and the minimum inhibitory concentration of Cd was found to be 1 mM. Strain G303 effectively remove 94.7 % of Cd after 96 h of culture. Extracellular CdS was detected using multiple methods, with the CdS formed being aggregated in the biofilm. The addition of cysteine and thiosulfate to the medium significantly enhanced the Cd resistance and removal capacity of strain G303. Integrated genomic and proteomic analyses revealed that heavy metal transporters cooperate to resist Cd stress. Cysteine and thiosulfate improved Cd tolerance in strain G303 by upregulating nitrogen and energy metabolism. Proteins associated with nitrate reduction likely played a pivotal role in cysteine and thiosulfate metabolism. Notably, cysteine synthase and the SUF system played crucial roles in CdS formation. This study systematically explored the impact of cysteine and thiosulfate on the Cd resistance of strain G303, deepening our understanding of the microbial response mechanism to heavy metals.

Generation of a TSC2 knockout embryonic stem cell line by CRISPR/Cas9 editing.

Tuberous Sclerosis Complex (TSC) is a severe developmental disorder with various clinical effects, primarily caused by TSC2 gene mutations, often involving loss of function(Henske,et al., 2016).To explore role of TSC2 in human heart development, we successfully developed a TSC2 knockout (TSC2-/-) human embryonic stem cells (hESCs) line using CRISPR/Cas9 gene editing. This TSC2-/- hESC line maintained a normal karyotype, expressed pluripotency markers strongly, and could differentiate into all three germ layers in vivo. This cell line will be a valuable tool for future research on the role of TSC2 in heart development.