Citrullinemia type II accompanied by mental derangement combined with multidrug resistance 3 decrease, case report.

Background: Here we report a rare case of citrullinemia type II (CTLN2) accompanied by mental derangement with a deficiency of multidrug resistance 3 (MDR3) in the liver. Case presentation: The clinical data of a 17-year-old girl were collected. Liver puncture was performed, and hepatic expression of MDR3 was determined by immunohistochemistry. Serum amino acids of the patient and her parents wwere determined by a chemical isotope labeling liquid chromatography-mass spectrometry (CIL LC-MS). Genetic mutations of ABCB4 and SLC25A13 were screened by whole-exome sequencing. Immunohistochemical analysis showed a remarkably lower expression of MDR3. Mutation in ABCB4 gene was not found and whole-exome sequencing revealed the SLC25A13 mutation 852-855 del. Elevated serum levels of citrulline, homocitrulline, and homoarginine in the patient and her mother were found. Conclusions: We reported a rare case of CTLN2 combined with MDR3 deficiency, without mutation of ABCB4. The link between MDR3 down-expression and CTLN2 warrants further investigation. Meanwhile, clinicians need to further rule out the possibility of CTLN2 if MDR3 decreases in adolescent patients with mental disorders and abnormal liver function.

Generation and metabolomic characterization of functional ductal organoids with biliary tree networks in decellularized liver scaffolds.

Developing functional ductal organoids (FDOs) is essential for liver regenerative medicine. We aimed to construct FDOs with biliary tree networks in rat decellularized liver scaffolds (DLSs) with primary cholangiocytes isolated from mouse bile ducts. The developed FDOs were dynamically characterized by functional assays and metabolomics for bioprocess clarification. FDOs were reconstructed in DLSs retaining native structure and bioactive factors with mouse primary cholangiocytes expressing enriched biomarkers. Morphological assessment showed that biliary tree-like structures gradually formed from day 3 to day 14. The cholangiocytes in FDOs maintained high viability and expressed 11 specific biomarkers. Basal-apical polarity was observed at day 14 with immunostaining for E-cadherin and acetylated α-tubulin. The rhodamine 123 transport assay and active collection of cholyl-lysyl-fluorescein exhibited the specific functions of bile secretion and transportation at day 14 compared to those in monolayer and hydrogel culture systems. The metabolomics analysis with 1075 peak pairs showed that serotonin, as a key molecule of the tryptophan metabolism pathway linked to biliary tree reconstruction, was specifically expressed in FDOs during the whole period of culture. Such FDOs with biliary tree networks and serotonin expression may be applied for disease modeling and drug screening, which paves the way for future clinical therapeutic applications.

Structural optimization of tetrahydroisoquinoline-hydroxamate hybrids as potent dual ERα degraders and HDAC inhibitors.

Both estrogen receptor α (ERα) and histone deacetylases (HDACs) are valid therapeutic targets for anticancer drug development. Combination therapies using diverse ERα antagonists or degraders and HDAC inhibitors have been proven effective in endocrine-resistant ER + breast cancers based on the crosstalk between ERα and HDAC pathway. In this study, we reported the optimization of a series of methoxyphenyl- or pyridinyl- substituted tetrahydroisoquinoline-hydroxamates, which were optimized from 31, a dual ERα degrader/HDAC inhibitor previously reported by our group. Most of the synthesized compounds displayed potent ERα degradation efficacy and antiproliferative activity. Among them, A04 demonstrated the best anti-proliferation activity (MCF-7 IC50 = 1.96 µM) and HDAC6 inhibitory activity (HDAC6 IC50 = 25.96 nM), which is slightly more potent than the lead compound 31 (MCF-7 IC50 = 4.38 µM, HDAC6 IC50 = 63.03 nM). In addition, compound A04 exerted ERα-independent HDAC6-inhibiting effect without agonistic activity in endometrial cells. These results demonstrated that A04 is a novel and promising dual ERα degrader/HDAC inhibitor worthy of further development.

Ocean acidificationf affects the bioenergetics of marine mussels as revealed by high-coverage quantitative metabolomics.

Ocean acidification has become a major ecological and environmental problem in the world, whereas the impact mechanism of ocean acidification in marine bivalves is not fully understood. Cellular energy allocation (CEA) approach and high-coverage metabolomic techniques were used to investigate the acidification effects on the energy metabolism of mussels. The thick shell mussels Mytilus coruscus were exposed to seawater pH 8.1 (control) and pH 7.7 (acidification) for 14 days and allowed to recover at pH 8.1 for 7 days. The levels of carbohydrates, lipids and proteins significantly decreased in the digestive glands of the mussels exposed to acidification. The 14-day acidification exposure increased the energy demands of mussels, resulting in increased electron transport system (ETS) activity and decreased cellular energy allocation (CEA). Significant carry-over effects were observed on all cellular energy parameters except the concentration of carbohydrates and cellular energy demand (Ec) after 7 days of recovery. Metabolomic analysis showed that acidification affected the phenylalanine, tyrosine and tryptophan biosynthesis, taurine and hypotaurine metabolism, and glycine, serine and threonine metabolism. Correlation analysis showed that mussel cell energy parameters (carbohydrates, lipids, proteins, CEA) were negatively/positively correlated with certain differentially abundant metabolites. Overall, the integrated biochemical and metabolomics analyses demonstrated the negative effects of acidification on energy metabolism at the cellular level and implicated the alteration of biosynthesis and metabolism of amino acids as a mechanism of metabolic perturbation caused by acidification in mussels.

Novel 6-amino-1,3,5-triazine derivatives as potent BTK inhibitors: structure-activity relationship (SAR) analysis and preliminary mechanism investigation.

Bruton's tyrosine kinase (BTK) is a promising drug target for the treatment of B-cell related malignancies. Irreversible inhibition of BTK by a covalent inhibitor has been proved to be a clinically effective therapy. However, most irreversible BTK inhibitors also inhibit other kinases including JAK3 and EGFR, leading to some adverse events. Herein, we reported the structure-based design and optimization of a series of irreversible BTK inhibitors bearing the 6-amino-1,3,5-triazine scaffold. Most of the synthesized compounds demonstrated considerable BTK inhibition and improved anti-proliferative activity against Raji and Ramos cells. Among them, compound C11 exhibited potent BTK inhibition (BTK IC50 = 17.0 nM) and a desirable selectivity profile especially over EGFR. Moreover, C11 effectively blocked activation of BTK and downstream signaling, arrested the cell cycle in G0/G1 phase and induced apoptosis in Raji cells. Its irreversible binding mode was further investigated by both molecular modeling and a washout experiment. Collectively, C11 is a novel selective irreversible BTK inhibitor worthy of further in-depth research.

Cavity-dumped burst-mode Nd:YAG laser master-oscillator power-amplifier system with a flat-top beam output realized by gain profile-controlled side pumping.

We report a compact cavity-dumped burst-mode Nd:YAG laser master-oscillator power-amplifier system with a flat-top intensity distribution across the output-beam section. Custom-designed gain profile-controlled diode side pumping modules providing flat-top and concave gain profiles were utilized to generate a uniform beam profile and suppress thermal lensing during amplification, respectively. Bursts with an energy of 2.0 J and duration of 1.6 ms were operated at 10 Hz. Within the bursts, single pulses with an energy of 12.7 mJ and pulse width of 3.3 ns were achieved at 100 kHz.

5 kHz, 4.2mJ, 900 ps end-pumped Nd:YVO4 MOPA laser system.

A 5 kHz sub-nanosecond master oscillator power amplifier (MOPA) laser system was reported in this paper. The master oscillator was an electro-optically Q-switched Nd:YVO4 laser directly pumped at 879 nm, yielding a pulse energy of 520 µJ and a pulse width of 900 ps at 5 kHz. With two Nd:YVO4 amplifiers directly pumped at 914 nm, the pulse energy was further scaled up. Under the absorbed pump energy of 11.0 mJ, the pulse energy was amplified to 4.2 mJ, corresponding to a peak power of 4.7 MW. The optical-to-optical efficiency of the amplifiers reached 33.5%.

Metabolic perturbations in human hepatocytes induced by bis (2-ethylhexyl)-2,3,4,5-tetrabromophthalate exposure: Insights from high-coverage quantitative metabolomics.

Bis (2-ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH) is an extensively used novel brominated flame retardant that is present ubiquitously in the environment and in biota. However, there is inadequate data on its potential hepatotoxicity to humans. In this study, high-coverage quantitative metabolomics based on 12C-/13C-dansylation labeling LC-MS was performed for the first time to assess the metabolic perturbations and underlying mechanisms of TBPH on human hepatocytes. HepG2 cells were exposed to TBPH at dosages of 0.1,1,10 µM for 24 or 72 h. Overall, 1887 and 1364 amine/phenol-containing metabolites were relatively quantified in cells and culture supernatant. Our results revealed that exposure to 0.1 µM TBPH showed little adverse effects, whereas exposure to 10 µM TBPH for 24 h enhanced intracellular protein catabolism and disrupted energy and lipid homeostasis-related pathways such as histidine metabolism, pantothenate and CoA biosynthesis, alanine, aspartate and glutamate metabolism. Nevertheless, most of these perturbations returned to the same levels as controls after 72 h of exposure. Additionally, prolonged TBPH exposure increased oxidative stress, as reflected by marked disturbances in taurine metabolism. This study sensitively revealed the dysregulations of intracellular and extracellular metabolome induced by TBPH, providing a comprehensive understanding of metabolic responses of cells to novel brominated flame retardants.

The Role of Dihydroresveratrol in Enhancing the Synergistic Effect of Ligilactobacillus salivarius Li01 and Resveratrol in Ameliorating Colitis in Mice.

Currently approved therapeutical strategies for inflammatory bowel diseases (IBD) suffer from variable efficacy and association with risk of serious side effects. Therefore, efforts have been made in searching for alternative therapeutics strategies utilizing gut microbiota manipulation. In this study, we show that the probiotic strain Ligilactobacillus salivarius Li01 (Li01) and the phytochemical prebiotic resveratrol (RSV) have synergistic effect in ameliorating colitis in mice. Oral coadministration of Li01 (109 CFU/d) and RSV (1.5 g/kg/d) promoted restoration of various inflammatory injuries and gut microbiota composition, exhibiting a favorable anti-inflammatory effect in DSS-induced colitis mice. The combination treatment was associated with reductions in the levels of proinflammatory cytokines IL-1ß and IL-6 and increases in the levels of the anti-inflammatory cytokine IL-17A in mouse serum. Moreover, the combination treatment was found to alter the composition and metabolism of the gut microbiota, especially influencing the production of short chain fatty acids and anti-inflammatory related molecules. The mechanism underlying the improved anti-inflammatory effect from the RSV and Li01 combination treatment was found to be associated with the environmental sensor mammalian aryl hydrocarbon receptor (AHR) and tryptophan metabolism pathway. Administration of RSV in combination with Li01 in different mouse model led to enhanced conversion of RSV into metabolites, including dihydroresveratrol (DHR), resveratrol-sulfate, and resveratrol-glucuronide. DHR was found to be the dominant metabolite of RSV in conventional and colitis mice. An increased DHR/RSV ratio was confirmed to activate AHR and contribute to an enhanced anti-inflammatory effect. DHR is considered as a potential AHR ligand. The DHR/RSV ratio also affected the serotonin pathway by controlling the expression of Tph1, SERT, and 5-HT7R leading to amelioration of colitis in mice. Our data suggest that treatment with a combination of Li01 and RSV has potential as a therapeutic strategy for IBD; further investigation of this combination in clinical settings is warranted.

Dynamic alterations of metabolites revealed the vascularization progression of bioengineered liver.

Vascularization is a critical but challenging process in developing functional bioengineered livers with the decellularized liver scaffolds (DLSs) and the process is accompanied by cell-specific metabolic alterations. To elucidate the dynamic alterations of metabolites during vascularization, rat DLSs were vascularized with human umbilical vein endothelial cells and liquid chromatography mass spectrometry-based metabolomics was performed on culture supernatants collected at 0, 1, 3, 7, 14, and 21 days. Overall, 1698 peak pairs or metabolites were detected in the culture supernatants, with 309 metabolites being positively identified. The orthogonal partial least-squares discriminant analysis and functional enrichment analysis revealed three phases that could be clearly discriminated, including Phase D1 (cell proliferation and migration), Phase D3D7 (vascular lumen formation), and Phase D14D21 (functional endothelial barrier formation). Seventy-two common differentially abundant metabolites of known identity were detected in these three phases when compared with Day 0. Of these metabolites, a high level of ß-Alanine indicated a better degree of vascularization and 14 days of in vitro dynamic culture is required to develop a functionalized vascular structure. These results enriched our understanding of the metabolic mechanism of DLS vascularization and indicated that ß-Alanine could function as a potential predictor of the patency of vascularized bioengineered livers.

High-Coverage Quantitative Metabolomics of Human Urine: Effects of Freeze-Thaw Cycles on the Urine Metabolome and Biomarker Discovery.

Urine sample storage after collection at ultra-low-temperature (e.g., -80 °C) is normally required for comparative metabolome analysis of many samples, and therefore, freeze-thaw cycles (FTCs) are unavoidable. However, the reported effects of FTCs on the urine metabolome are controversial. Moreover, there is no report on the study of how urine FTCs affect biomarker discovery. Herein, we present our study of the FTC effects on the urine metabolome and biomarker discovery using a high-coverage quantitative metabolomics platform. Our study involved two centers located in Hangzhou, China, and Edmonton, Canada, to perform metabolome analysis of two separate cohorts of urine samples. The same workflow of sample preparation and dansylation isotope labeling LC-MS was used for in-depth analysis of the amine/phenol submetabolome. The analysis of 320 samples from the Hangzhou cohort consisting of 80 healthy subjects with each urine being subjected to four FTCs resulted in relative quantification of 3682 metabolites with 3307 identified or mass-matched. The analysis of 176 samples from the Edmonton cohort of 44 subjects with four FTCs quantified 3516 metabolites with 3166 identified or mass-matched. Multivariate and univariate analyses indicated that significant variations (fold change ≥ 1.5 with q-value ≤ 0.05) from FTCs were only observed in a very small fraction of the metabolites (<0.3%). Moreover, various metabolites did not show a consistent pattern of concentration changes from one to four FTCs, allowing the use of two separate cohorts of samples to remove these randomly changed metabolites. Three metabolite biomarkers for separating males and females were discovered, and FTC did not influence their discovery.

Building roof extraction from ASTIL echo images applying OSA-YOLOv5s.

Light detection and ranging (LiDAR) is a type of essential tool for urban planning and geoinformation extraction. Airborne streak tube imaging LiDAR (ASTIL) is a new system with great advantages in the rapid collection of remote sensing data. To the best of our knowledge, a new method to extract a building roof from the echo images of ASTIL is proposed. We improve YOLOv5s with a one-shot aggregation (OSA) module to improve efficiency. The experimental results show that the mean average precision of the OSA-YOLOv5s algorithm can reach 95.2%, and the frames per second can reach 11.74 using a CPU and 39.39 using a GPU. The method proposed can extract building objects efficiently from the echo images of ASTIL and acquire the building roof point cloud.

Generation and amplification of a multi-beam sub-nanosecond laser.

In this paper, a methodology to produce a multi-beam sub-nanosecond laser is proposed. Laser pulses with a pulse energy of 0.14 mJ and a pulse width of 490 ps are generated in a YAG/Nd:YAG/Cr4+:YAG microchip laser at a repetition rate of 200 Hz. After amplification with a laser diode (LD) side-pumped Nd:YAG module, four laser beams are generated because of the thermally induced birefringence. With a double-pass LD side-pumped amplifier, the single pulse energy of the four laser beams is amplified to 5.23 mJ with a peak power of ∼10.67 MW, and air breakdown with four points is achieved with a 2 × 2 lens array.

Metabolic profile of irradiated whole blood by chemical isotope-labeling liquid chromatography-mass spectrometry.

Irradiated blood is a new type of blood product used to prevent transfusion-associated graft-versus-host disease. However, the effects of irradiation on the metabolism of plasma, red blood cells (RBCs), and peripheral blood mononuclear cells (PBMCs) are largely unknown. We developed a workflow for testing metabolic changes in whole blood to determine the impact of irradiation by chemical isotope labeling liquid chromatography-mass spectrometry (CIL LC-MS). Blood parameters, PBMC proliferation and apoptosis were examined before and after irradiation. Next, the amine/phenol metabolites in the blood components were assayed by 12C- and13C-dansylation labeling LC-MS. We identified 1654, 1730, and 1666 peak pairs in plasma, RBCs, and PBMCs, respectively. We screened out 367, 177, and 219 significant metabolites in plasma, RBCs, and PBMCs, respectively, by principle component analyses, volcano plots, and Venn plots. Metabolic pathway analyses showed that irradiation modulated taurine and hypotaurine metabolism in plasma and purine metabolism in RBCs and PBMCs. Changes in potential biomarkers, including an increase in hypoxanthine level and a decrease in adenine level, may be related to the dysfunction of DNA synthesis in PBMCs. The decreased AMP level in RBCs may interfere with RBC storage lesions. Our research provides a more comprehensive perspective on blood metabolism associated with irradiation.

Device-Free Human Activity Recognition with Low-Resolution Infrared Array Sensor Using Long Short-Term Memory Neural Network.

Sensor-based human activity recognition (HAR) has attracted enormous interests due to its wide applications in the Internet of Things (IoT), smart homes and healthcare. In this paper, a low-resolution infrared array sensor-based HAR approach is proposed using the deep learning framework. The device-free sensing system leverages the infrared array sensor of 8×8 pixels to collect the infrared signals, which can ensure users' privacy and effectively reduce the deployment cost of the network. To reduce the influence of temperature variations, a combination of the J-filter noise reduction method and the Butterworth filter is performed to preprocess the infrared signals. Long short-term memory (LSTM), a representative recurrent neural network, is utilized to automatically extract characteristics from the infrared signal and build the recognition model. In addition, the real-time HAR interface is designed by embedding the LSTM model. Experimental results show that the typical daily activities can be classified with the recognition accuracy of 98.287%. The proposed approach yields a better result compared to the existing machine learning methods, and it provides a low-cost yet promising solution for privacy-preserving scenarios.

High-pulse-energy passively Q-switched sub-nanosecond MOPA laser system operating at kHz level.

A passively Q-switched sub-nanosecond master oscillator power amplifier (MOPA) laser system at 1064 nm has been reported in this paper. The master oscillator was a passively Q-switched YAG/Nd:YAG/Cr4+:YAG microchip laser, yielding a pulse energy of 0.14 mJ and a pulse width of ∼490 ps at repetition rates of 500 Hz and 1 kHz. After passing a double-pass side-pumped Nd:YAG amplification system, the pulse energy reached 7.6 mJ and 1.7 mJ at 500 Hz and 1 kHz, respectively. The spatial beam deformation caused by the thermally induced birefringence was investigated numerically and experimentally.

Toxicity mechanisms of polystyrene microplastics in marine mussels revealed by high-coverage quantitative metabolomics using chemical isotope labeling liquid chromatography mass spectrometry.

Marine microplastic has become an important environmental issue of global concern due to its wide distribution and harmful impacts. However, there is still insufficient information on the toxicity mechanism of microplastics to marine organisms. In this study, we developed and applied a high-coverage quantitative metabolomics technique to investigate the toxicity mechanisms of the polystyrene microspheres (micro-PS) on marine mussels (Mytilus coruscus). A total of 3599 metabolites were quantified, including 163 positively identified metabolites, 318 high-confident putatively identified metabolites, and 2602 mass-matched metabolites from the hemolymph of mussels. Metabolomics analysis indicated that micro-PS disrupted the amino acid metabolism, particularly phenylalanine metabolism, which may lead to oxidative stress and neurotoxicity. Micro-PS at environmentally relevant concentrations induced oxidative stress and immunotoxicity in mussels. After 7 days of recovery, along with the significant clearance of micro-PS by mussels, both metabolite levels and biochemical indicators generally returned to the same level as the control group. Overall, the results showed that microplastics at environmentally-relevant concentrations can cause toxic effects on mussels but these influences are reversible. We envisage the usages of high-coverage metabolomics for investigating the toxicity of various types of microplastics under many different conditions, including those relevant to the marine environment.

High-Coverage Metabolome Analysis Reveals Significant Diet Effects of Autoclaved and Irradiated Feed on Mouse Fecal and Urine Metabolomics.

SCOPE: Using metabolomics to study the relations of nutrition and health requires stringent control of the experimental conditions used in an animal model. This work investigates the diet effects of autoclaved and irradiated feed on mouse urine and fecal metabolomics. METHODS AND RESULTS: C57BL/6 mice are fed normal-irradiation sterilized diet (n = 9), autoclave sterilized diet (n = 9), and high-irradiation sterilized diet (n = 9) for 4 weeks. Differential chemical isotope labeling liquid chromatography mass spectrometry is used to quantify the metabolome variations of urine and feces collected at five time points. Significant differences are observed in urine or fecal metabolomes of mice fed autoclaved diet versus mice fed high-irradiation diet or fed normal-irradiation diet, while the differences are small between the mice fed normal-irradiation and high-irradiation diet. Correlation studies of metabolite changes of diet- and aging-related biomarkers indicate a large overlap of significantly affected metabolites by the two factors. CONCLUSIONS: Diet can be a confounding factor that needs to be carefully considered when a metabolomics study is designed and metabolomic results of a mouse model of nutritional or other biological study are interpreted. Using the same sterilized diet for a given metabolomics project is essential to control the diet effect.

High-coverage quantitative liver metabolomics using perfused and non-perfused liver tissues.

Comprehensive analysis of the liver metabolome can be very useful for discovering disease biomarkers and studying diseases, especially liver-related diseases. However, the presence of a relatively large amount of blood in liver tissue may have a profound effect on liver tissue metabolome analysis. We designed a study to address this issue in order to develop a liver metabolomics workflow based on high-coverage quantitative metabolome analysis using differential chemical isotope labeling (CIL) LC-MS. In the first set of experiments, we compared the metabolomes of mouse serum, non-perfused liver, and perfused liver without and with varying amounts of blood added. We found that there was a significant metabolome difference between the perfused liver and non-perfused liver. To illustrate the effects of perfusion conditions on tissue metabolome analysis, we analyzed the mouse livers that were subjected to perfusion under two different conditions. We found that ice-cold temperature perfusion led to less change of the liver metabolome, compared to room temperature perfusion; however, there was still a significant metabolome difference between the ice-cold-perfused liver and the non-perfused liver. Finally, we applied the method to a chemical (carbon tetrachloride) exposure liver injury model to examine the effects of blood in liver on the detection of significantly changed metabolites in two comparative groups of mice. Using multivariate and univariate analyses of the serum and liver metabolomes of control and diseased mice, we detected many unique significant metabolites in serum as well as in liver. This work demonstrates that perfusion can alter the liver metabolome significantly. Therefore, we recommend the use of non-perfused liver for high-coverage liver metabolomics.

GC-MS metabolomics identifies novel biomarkers to distinguish tuberculosis pleural effusion from malignant pleural effusion.

BACKGROUND: Tuberculous pleural effusions (TBPEs) and malignant pleural effusions (MPEs) are two of the most common and severe forms of exudative effusions. Clinical differentiation is challenging; however, metabolomics is a collection of powerful tools currently being used to screen for disease-specific biomarkers. METHODS: 17 TBPE and 17 MPE patients were enrolled according to the inclusion criteria. The normalization gas chromatography-mass spectrometry (GC-MS) data were imported into the SIMCA-P + 14.1 software for multivariate analysis. The principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) were used to analyze the data, and the top 50 metabolites of variable importance projection (VIP) were obtained. Metabolites were qualitatively analyzed using the National Institute of Standards and Technology (NIST) databases. Pathway analysis was performed by MetaboAnalyst 4.0. The detection of biochemical indexes such as urea and free fatty acids in these pleural effusions was also verified, and significant differences were found between these two groups. RESULTS: 1319 metabolites were screened by non-targeted metabonomics of GC-MS. 9 small molecules (urea, L-5-oxoproline, L-valine, DL-ornithine, glycine, L-cystine, citric acid, stearic acid, and oleamide) were found to be significantly different (p < 0.05 for all). In OPLS-DA, 9 variables were considered significant for biological interpretation (VIP≥1). However, after the ROC curve was performed, it was found that the metabolites with better diagnostic value were stearic acid, L-cystine, citric acid, free fatty acid, and creatinine (AUC > 0.8), with good sensitivity and specificity. CONCLUSION: Stearic acid, L-cystine, and citric acid may be potential biomarkers, which can be used to distinguish between the TBPE and the MPE.