Development of Transmission Ambient Pressure Laser Desorption Ionization/Postphotoionization Mass Spectrometry Imaging.

Laser-based high-resolution mass spectrometry imaging at ambient conditions has promising applications in life science. However, the ion yield during laser desorption/ablation is poor. Here, transmission atmospheric pressure laser desorption ionization combined with a compact postphotoionization (t-AP-LDI/PI) assembly with a krypton discharge lamp was developed for the untargeted imaging of various biomolecules. The spatial distributions of numerous lipid classes, fatty acids, neurotransmitters, and amino acids in the subregions of mouse cerebellum tissue were obtained. Compared with single laser ablation, the sensitivities for most analytes were increased by 1 to 3 orders of magnitude by dopant-assisted postphotoionization. After careful optimization, a spatial resolution of 4 µm could be achieved for the metabolites in mouse hippocampus tissue. Finally, the melanoma tissue slices were analyzed using t-AP-LDI/PI MSI, which revealed the metabolic heterogeneity of the melanoma microenvironment and exhibited the phenomenon of abnormal proliferation and invasion trends in tumor cells.

Enhanced imaging of endogenous metabolites by negative ammonia assisted DESI/PI mass spectrometry.

In this work, endogenous metabolites in mouse brain tissue were imaged by negative desorption electrospray ionization (DESI), ammonia assisted DESI (aa-DESI), DESI/post-photoionization (DESI/PI), and ammonia assisted DESI/PI (aa-DESI/PI) mass spectrometry imaging (MSI) strategies. The combined effect of ammonia additive and post-photoionization was found to play an important role in the enhancement of sensitivity and coverage for endogenous analytes under ambient conditions. Compared with DESI, aa-DESI/PI can provide increased signal intensities for metabolites up to 37.1-fold, as well as the imaging of nine more small metabolites (m/z < 350) (26 for aa-DESI/PI and 17 for DESI) in mouse brain tissue. The results of Pearson correlation analysis and KEGG pathway analysis showed that the enhanced imaging strategy of aa-DESI/PI can facilitate the study of endogenous metabolic pathways and molecular networks. Moreover, the imaging results of mouse tumor tissue demonstrated the promising application of aa-DESI/PI in tumor research.

Online Monitoring the Key Intermediates and Volatile Compounds Evolved from Green Tea Roasting by Synchrotron Radiation Photoionization Mass Spectrometry.

Online monitoring of the volatile compounds during the tea roasting process is crucial to find the optimum roasting conditions and improve the quality of green tea. In this work, synchrotron radiation photoionization mass spectrometry (SR-PIMS) was utilized to online monitor the evolved gaseous compounds during the tea roasting process. By virtue of "soft" ionization and fast data acquisition characteristics of SR-PIMS, dozens of aroma compounds including alcohols, aldehydes, furans, and nitrogen- and sulfur-containing species were detected and identified in real time. Moreover, 5-hydroxymethylfurfural (5-HMF), the key intermediate of Maillard reactions, was found with high sensitivity. Evolution processes of all the products could be observed via the time- and temperature-resolved profiles in N2 and the air. Dehydration was found to be the first step during roasting. Oxygen in the air was found to accelerate the formation rate of various stable species and intermediates in the course of the thermal treatment of fresh green tea. The formation mechanisms of evolved compounds such as three sulfur-containing compounds, i.e., dimethyl sulfide, hydrogen sulfide, and methanethiol, could be proposed according to the step-by-step formation process. The time-resolved results were demonstrated to be applicable in the evaluation of different roasting processes by statistical analysis. The optimum tea roasting temperature and duration are proposed to be around 200 °C and 1000 s.

Cholesterol was identified as a biomarker in human melanocytic nevi using DESI and DESI/PI mass spectrometry imaging.

The rapid differentiation between diseased tissue and healthy normal tissue is of great importance for the intraoperative diagnosis. Herein, desorption electrospray ionization (DESI) and DESI/post-photoionization (DESI/PI) mass spectrometry imaging were combined to in situ visualize the distribution of biochemicals within the tissue regions of human melanocytic nevi under the ambient condition with a spatial resolution of around 200 µm. Plenty of polar and nonpolar lipids were found to be specifically distributed in melanocytic nevi with statistical significance and could be used to differentiate the healthy normal tissue and melanocytic nevi. Cholesterol was further confirmed to be a potential biomarker for melanocytic nevi diagnosis by multivariate statistical analysis and immunohistochemistry of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) and translocator protein (TSPO) enzymes. This work provides a visual way for the diagnosis of human melanocytic nevi by lipid profiling, which benefits the understanding of the pathological mechanism of melanocytic nevi and provides a new insight to control melanin growth from the synthesis, transport, and metabolism of cholesterol.

Fast and comprehensive characterization of chemical ingredients in traditional Chinese herbal medicines by extractive atmospheric pressure photoionization (EAPPI) mass spectrometry.

RATIONALE: The goal of this work is to employ extractive atmospheric pressure photoionization mass spectrometry (EAPPI-MS) to characterize the constituents in traditional Chinese herbal medicine (TCHM) directly without chromatographic separation. METHODS: Sample was placed in 4 mL of methanol/water (v/v, 3:1) in the nebulization cell, and then the ultrasonic nebulizer was switched on. The ultrasonic nebulization system allows the simultaneous sample extraction and introduction of extract aerosols. The extract aerosols were vaporized in a transfer tube. Mixed with a gaseous dopant, vaporized analytes were ionized through ambient photon-induced ion-molecule reactions, and were mass-analyzed by high-resolution time-of-flight mass spectrometry (TOF-MS). RESULTS: The major ingredients including alkaloids, flavonoids, amino acids, saccarides, ginsenosides, lignans and terpenoids were readily detected. Compared with electrospray ionization (ESI), EAPPI allowed the ionization of a wider range of compounds, which is desirable for the integral characterization of TCHMs containing numerous constituents. The significant discrepancies for both alkaloids and terpenoids in tripterygium glycoside tablets from two different manufacturers could be simultaneously reflected from EAPPI mass spectra. CONCLUSIONS: Our results demonstrate that EAPPI-MS can be regarded as a supplementary ambient method for the fast and comprehensive analysis of TCHMs, which is important for the quality control and safety assurance of these products.

Therapeutic effects of different doses of polyethylene glycosylated porcine glucagon-like peptide-2 on ulcerative colitis in male rats.

BACKGROUND: Polyethylene glycosylated (PEGylated) porcine glucagon-like peptide-2 (pGLP-2) considerably increases half-life and stability compared with the native pGLP-2, but the effective dose for intestinal damage is still unclear. This study aims to evaluate the available dose of polyethylene glycosylated porcine glucagon-like peptide-2 (PEG-pGLP-2), a modified, long-acting form of pGLP-2 in an experimental rat model of ulcerative colitis. METHODS: Thirty-five male rats were randomly assigned into five groups: control, dextran sodium sulphate (DSS), DSS + PEG-pGLP-2(L), DSS + PEG-pGLP-2(M) and DSS + PEG-pGLP-2(H). Rats in control group received only water; other rats were fed with 5% (w/v) DSS and intraperitoneally administered with 12.5, 25 and 100 nmol/kg PEG-pGLP-2 daily for 6 days. RESULTS: Compared with the control treatment, DSS treatment significantly (p < 0.05) decreased body weight change, colonic length, duodenal villus height and expression of zonula occludens-1, whereas significantly (p < 0.05) increased colonic damage score and expression of claudin-1, interleukin (IL)-1, IL-7, IL-10, interferon-γ and tumour necrosis factor (TNF)-α in colon. However, the three doses of PEG-pGLP-2 all reduced these effects; these treatments significantly (p < 0.05) increased body weight change and duodenal villus height, whereas significantly (p < 0.05) decreased colonic damage score and expression of IL-1, IL-7 and TNF-α in colon. Specifically, low-dose (12.5 nmol/kg/d) PEG-pGLP-2 was effective. CONCLUSIONS: These results indicated that PEG-pGLP-2 is a novel and potentially effective therapy for intestinal healing in a relatively low dose.

Glucagon-like peptide-2-loaded microspheres as treatment for ulcerative colitis in the murine model.

Glucagon-like peptide-2 (GLP-2) is an intestinal hormone that promotes intestinal growth, but the rapid degradation by dipeptidyl peptidase-IV limits its applications. PLGA microsphere is a well-developed drug delivery system, while seldom been studied as a solution for prolonging in vivo effects of GLP-2. In this study, we encapsulated porcine GLP-2 (pGLP-2) into microspheres and investigated its therapeutic effects in dextran sulfate sodium (DSS)-treated mice. pGLP-2 microspheres showed 20.36% in initial burst and constant release for at least 9 d. In the DSS-treated mice, a single injection of GLP-2 microspheres significantly increased the body weight, colonic length, small intestinal weight and mRNA expression of Occludin, decreased the colonic damage score, mRNA expression of IL-6, IL-10, TNF-α and IFN-γ. In conclusion, pGLP-2 microspheres were resistant to degradation and decreased the severity of DSS-induced ulcerative colitis which suggested that GLP-2-loaded microspheres could be a proper candidate for the treatment of ulcerative colitis.

Effects of PEGylated porcine glucagon-like peptide-2 therapy in weaning piglets challenged with lipopolysaccharide.

This study aims to evaluate the therapeutic effect of polyethylene glycosylated porcine glucagon-like peptide-2 (pGLP-2), a long-acting form of pGLP-2, in lipopolysaccharide (LPS)-challenged piglets. Eighteen 21-day-old weaning piglets were randomly assigned into three groups: control (saline solution), LPS (100 µg/kg LPS), and PEG-pGLP-2 (10 nmol/kg PEG-pGLP-2+100 µg/kg LPS). All treatments were administered intraperitoneally. Compared with the control treatment, LPS treatment significantly decreased (P<0.05) the villus heights of the duodenum and jejunum, as well as the villus height/crypt depth ratio of the jejunum. However, PEG-pGLP-2 therapy reduced these effects (P>0.05). Specifically, PEG-pGLP-2 infusion significantly increased the villus height/crypt depth ratio of the duodenum (P<0.05) compared with LPS treatment. Compared with the control treatment, LPS treatment significantly increased (P<0.05) the mRNA expression levels of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the jejunum. However, PEG-pGLP-2 therapy reduced these effects (P<0.05). Specifically, PEG-pGLP-2 infusion significantly decreased (P<0.05) the mRNA expression levels of interleukin (IL)-8 and TNF-α in the duodenum and jejunum, IL-10 in the duodenum, and IFN-γ in the jejunum compared with the LPS treatment. LPS treatment increased the caspase-3 activity of the ileum mucosal (P<0.05), and this effect was significantly reduced by PEG-pGLP-2 treatment. These results indicate that PEG-pGLP-2 infusion alleviates the severity of intestinal injury in weaning piglets by reducing the secretion of inflammatory cytokines and the caspase-3 activity, and increasing the villus height/crypt depth ratio.