Untargeted metabolomics uncovers metabolic dysregulation and tissue sensitivity in ACE2 knockout mice.

Angiotensin-converting enzyme 2 (ACE2) polymorphisms are associated with increased risk of type 2 diabetes mellitus (T2DM), obesity and dyslipidemia, which have been determined in various populations. Consistently, ACE2 knockout (ACE2 KO) mice display damaged energy metabolism in multiple tissues, especially the key metabolic tissues such as liver, skeletal muscle and epididymal white adipose tissue (eWAT) and show even more severe phenotype under high-fat diet (HFD) induced metabolic stress. However, the effects of ACE2 on global metabolomics profiling and the tissue sensitivity remain unclear. To understand how tissues independently and collectively respond to ACE2, we performed untargeted metabolomics in serum in ACE2 KO and control wild type (WT) mice both on normal diet (ND) and HFD, and in three key metabolic tissues (liver, skeletal muscle and eWAT) after HFD treatment. The results showed significant alterations in metabolic profiling in ACE2 KO mice. We identified 275 and 168 serum metabolites differing significantly between WT and ACE2 KO mice fed on ND and HFD, respectively. And the altered metabolites in the ACE2 KO group varied from 90 to 196 in liver, muscle and eWAT. The alterations in ND and HFD serum were most similar. Compared with WT mice, ACE2 KO mice showed an increase in N-phenylacetylglutamine (PAGln), methyl indole-3-acetate, 5-hydroxytryptophol, cholic acid, deoxycholic acid and 12(S)-HETE, while LPC (19:0) and LPE (16:1) decreased. Moreover, LPC (20:0), LPC (20:1) and PC (14:0e/6:0) were reduced in both ND and HFD serum, paralleling the decreases identified in HFD skeletal muscle. Interestingly, DL-tryptophan, indole and Gly-Phe decreased in both ND and HFD serum but were elevated in HFD liver of ACE2 KO mice. A low level of l-ergothioneine was observed among liver, muscle, and epididymal fat tissue of ACE2 KO mice. Pathway analysis demonstrated that different tissues exhibited different dysregulated metabolic pathways. In conclusion, these results revealed that ACE2 deficiency leads to an overall state of metabolic distress, which may provide a new insight into the underlying pathogenesis in metabolic disorders in both ACE2 KO mice and in patients with certain genetic variant of ACE2 gene.

KIT A502_Y503 duplication mutation serves as a potential and universal target for neoantigen peptide in Chinese GIST patients.

BACKGROUND AND AIM: Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor with high prevalence of KIT and PDGFRA mutations. Few effective treatments can be exploited in imatinib or sunitinib resistant cases. While in immunotherapy, application of the highly individualized cancer neoantigen vaccines is hampered due to high economic and time cost. In this study we identified the most frequent mutation in Chinese GIST patients and predicted candidate neopeptide by next generation sequencing (NGS). METHODS: Tumor tissues and matched blood samples of 116 Chinese GIST patients were collected. Genomic profile was detected through NGS, and 450 cancer genes were deeply sequenced. KIT mutations were identified, and long peptides containing the mutation were queried in NetMHCpan 4.0 tools to predict MHC class I binding of mutant peptides. RESULTS: The most frequent mutated genes in detected GIST patients were KIT (81.9%, 95/116), CDKN2A (18.97%, 22/116), and CDKN2B (15.52%, 18/116) in this cohort. The most common mutation of KIT was A502_Y503 duplication (15.93%, 18/113) in exon 9. Among the 116 cases, 103 were HLA I genotyped, and 101 were HLA II genotyped. In total, 16 samples with the mutation of KIT p.A502_Y503dup were identified to produce neoantigens with qualified HLA affinity. CONCLUSIONS: KIT hotspot mutation (p.A502_Y503dup) has the highest incidence, which may further eliminate the need for whole genome sequencing and patient-specific neoantigen prediction and synthesis. Therefore, for those carrying such mutation, accounting for around 16% of Chinese GIST patients and are usually less sensitive to imatinib, effective immunotherapies are in prospect.

A multi-responsive indium-viologen hybrid with ultrafast-response photochromism and electrochromism.

A novel 0D organic-inorganic metal halide hybrid (C13H16N2O2)2InCl6·Cl (1) has been obtained by integrating the mono-viologen derivative with InCl3. Compound 1 exhibits reversible and ultrafast UV/sunlight/X-ray induced photochromic properties, as well as excellent electrochromic performance, which is the first example of an indium-based organic-inorganic chromic hybrid.

Heterogeneity response to afatinib in gastric cancer patient with uncommon epidermal growth factor receptor (EGFR) mutations: a case report.

Despite concerted efforts that have been made to characterize and understand the genomic landscape of gastric cancer (GC), only HER2 has been validated as a molecular target for GC treatment. Identifying new valid therapeutic targets is important for the treatment of this disease. The present report describes a Chinese male with a history of smoking two packs per day, who did not have a family history of cancer or other hereditary diseases, we discovered a small painless lump in the right groin in February 2018. Histopathology revealed a primary gastric adenocarcinoma. Positron emission tomography-computed tomography (PET-CT) showed multiple hypermetabolic nodules in the right upper lung, greater curvature of the stomach, and muscles. The patient had received treatment included oxaliplatin, docetaxel, and tegafur for two cycles, and second-line therapy of irinotecan and capecitabine, inguinal mass excision followed by concurrent radio-chemotherapy. However, the disease rapidly progressed. Whole exome sequencing (WES) showed uncommon epidermal growth factor receptor (EGFR) mutation of G719S + L861Q. The following EGFR tyrosine kinase inhibitors (TKIs) afatinib demonstrated partial response. Two months after targeted therapy, gastroscopy indicated rapid progression. With subsequent gastric specimen WES analysis, secondary MET amplification was found. The patient received local radiotherapy for gastric lesions as well as oral administration of apatinib. However, the disease rapidly progressed. A month later, he died of hepatic encephalopathy caused by obstructive jaundice combined with pulmonary and biliary tract infection. The present study indicated that afatinib might be a beneficial therapeutic option for a subset of GC patients with rare EGFR mutation in their tumors.

Formation of soy protein isolate-carrageenan complex coacervates for improved viability of Bifidobacterium longum during pasteurization and in vitro digestion.

Soy protein isolate (SPI) and carrageenan (IC) complex coacervates were formed through electrostatic attractions for encapsulating Bifidobacterium longum. The effects of pH (2.0-5.0) and SPI:IC mass ratios (10:1, 15:1, 20:1) on coacervate yield, entrapment efficiency and viability of the probiotic bacteria were investigated. The coacervates produced at pH 3 had higher yields and entrapment efficiency, and a SPI:IC mass ratio of 10:1 produced a complex coacervate with more compact microstructure. Compared to the native ones, the bacteria encapsulated in the coacervates had significantly improved viability during storage (4 °C), pasteurization (85 °C for 5, 10 and 30 min) and in vitro dynamic gastric and intestinal digestion. The findings also suggested that the coacervate with a SPI:IC ratio of 10:1 was more capable to protect the bacteria from loss against different stresses. This study provides a novel approach for designing efficient microcapsules containing probiotic bacteria with enhanced functional properties.

Total Phospholipids in Edible Oils by In-Vial Solvent Extraction Coupled with FTIR Analysis.

A simple procedure for the determination of total phospholipids (TPL) in edible oils was developed by combining a single-step, in situ methanol/acetonitrile (MeOH/ACN) extraction of the oil sample followed by Fourier transform infrared (FTIR) spectroscopic analysis of the extract. Spectral analysis of extracts in a 25 µm CaF2 cell obtained using 1:1 MeOH/ACN added to oil in a 2:1 ratio indicated that measurements made using only the asymmetric phosphate diester PO2- stretching band at 1243 cm-1 in second-derivative spectra were sufficient for the accurate measurement of TPL with minimal coextracted triglyceride interferences being encountered. FTIR calibration spectra were devised using only phosphatidylcholine (PC) as a representative phospholipid standard, covering a range of 0-50000 µg/g TPL and spiked into 1:1 MeOH/ACN, capable of tracking the added PC with an SD of <200 µg/g. The FTIR method was initially validated using model PC-spiked degummed canola oil and subsequently with commercial crude and refined soy and rapeseed oils as well as a lecithin tablet with the FTIR TPL predictions compared to those of the AOCS Ca 12-55 molybdenate method. The FTIR method tracked the AOCS results well, being somewhat more reproducible than the reference method (±3.2 vs ±4.9%), which limited its accuracy relative to the AOCS reference procedure (±2.2%). The simple in-vial solvent extraction procedure, followed by FTIR analysis of the extract, is a simple, efficient, and rapid procedure that is also amenable to automation using an autosampler-equipped FTIR if multiple samples are to be analyzed.

Rapid determination of phospholipid content of vegetable oils by FTIR spectroscopy combined with partial least-square regression.

A rapid mid-FTIR method was developed to quantitatively determine the total phospholipid (PL) content of vegetable oils. The method simply requires that the oil be diluted 4:1 (w/w) with hexane, its spectrum taken and ratioed against a hexane background. A calibration was devised using partial least squares by adding purified soybean PL at levels of 0.02-2.0% to phospholipid-free oils (soybean, rapeseed, sunflower) using the spectral region encompassing 1,357-1,000 cm(-1) and validated using the AOCS 12-55. Using calibration and leave-one-out cross-validation predictive errors, a 200-20,000 ppm calibration was accurate to within ± 362 and 488 ppm, respectively, while for sub-calibrations ranging from 200 to 2000; 2000 to 8000 and 8000 to 20,000 ppm, they were ± 72-172, ± 119-220, and ± 242-371 ppm, respectively. Although limited to 3 oil types in this study, the calibration is simple to devise and can be broadened to the universe of oil types of interest, the analytical protocol being straightforward and the analysis readily automatable.

The aggregation of soy protein isolate on the surface of Bifidobacterium.

In this study the interaction between soy protein isolate (SPI) and Bifidobacterium was investigated to reveal the protecting mechanism of SPI on Bifidobacterium. The results indicated that the average particle size of the aggregate of SPI and Bifidobacterium longum (B. longum, the better surface hydrophobicity of four Bifidobacterium) exhibited more considerable expansion than SPI alone, from 295.9nm to 404.8nm (the aggregate). It was confirmed that B. longum was localized in the microparticle core, while SPI acted as a wall-coating material, as determined by zeta potential and laser scanning confocal microscopy (LSCM). Surface hydrophobicity, which was described by fluorescence intensity, of the aggregate of SPI and B. longum was decreased to 72% of SPI. The main binding force of the interaction between SPI and B. longum originated from a hydrophobic interaction was verified by isothermal titration calorimetry (ITC). The enthalpy (ΔH) was determined by ITC to be -3.24kJmol-1 for the adsorption of SPI on B. longum at 25°C and pH7.0. Furthermore, the aggregation was testified to be an endothermic process, and the process was spontaneous and irreversible. The hydrophobic forces between SPI and Bifidobacterium interpreted that SPI has the potential to be a useful food ingredient in protecting probiotics.