Phosphodiesterase 8 (PDE8): Distribution and Cellular Expression and Association with Alzheimer's Disease.

Phosphodiesterase 8 (PDE8), as a member of PDE superfamily, specifically promotes the hydrolysis and degradation of intracellular cyclic adenosine monophosphate (cAMP), which may be associated with pathogenesis of Alzheimer's disease (AD). However, little is currently known about potential role in the central nervous system (CNS). Here we investigated the distribution and expression of PDE8 in brain of mouse, which we believe can provide evidence for studying the role of PDE8 in CNS and the relationship between PDE8 and AD. Here, C57BL/6J mice were used to observe the distribution patterns of two subtypes of PDE8, PDE8A and PDE8B, in different sexes in vivo by western blot (WB). Meanwhile, C57BL/6J mice were also used to demonstrate the distribution pattern of PDE8 in selected brain regions and localization in neural cells by WB and multiplex immunofluorescence staining. Furthermore, the triple transgenic (3×Tg-AD) mice and wild type (WT) mice of different ages were used to investigate the changes of PDE8 expression in the hippocampus and cerebral cortex during the progression of AD. PDE8 was found to be widely expressed in multiple tissues and organs including heart, kidney, stomach, brain, and liver, spleen, intestines, and uterus, with differences in expression levels between the two subtypes of PDE8A and PDE8B, as well as two sexes. Meanwhile, PDE8 was widely distributed in the brain, especially in areas closely related to cognitive function such as cerebellum, striatum, amygdala, cerebral cortex, and hippocampus, without differences between sexes. Furthermore, PDE8A was found to be expressed in neuronal cells, microglia and astrocytes, while PDE8B is only expressed in neuronal cells and microglia. PDE8A expression in the hippocampus of both female and male 3×Tg-AD mice was gradually increased with ages and PDE8B expression was upregulated only in cerebral cortex of female 3×Tg-AD mice with ages. However, the expression of PDE8A and PDE8B was apparently increased in both cerebral cortex and hippocampus in both female and male 10-month-old 3×Tg-AD mice compared WT mice. These results suggest that PDE8 may be associated with the progression of AD and is a potential target for its prevention and treatment in the future.

Discovery of a novel chemotype as DYRK1A inhibitors against Alzheimer's disease: Computational modeling and biological evaluation.

Dual-specificity tyrosine phosphorylation-regulated kinase 1 A (DYRK1A) plays an essential role in Tau and Aß pathology closely related to Alzheimer's disease (AD). Accumulative evidence has demonstrated DYRK1A inhibition is able to reduce the pathological features of AD. Nevertheless, there is no approved DYRK1A inhibitor for clinical use as anti-AD therapy. This is somewhat due to the lack of effective and safe chemotypes of DYRK1A inhibitors. To address this issue, we carried out in silico screening, in vitro assays and in vivo efficacy evaluation with the aim to discover a new class of DYRK1A inhibitors for potential treatment of AD. By in silico screening, we selected and purchased 16 potential DYRK1A inhibitors from the Specs chemical library. Among them, compound Q17 (Specs ID: AO-476/40829177) potently inhibited DYRK1A. The hydrogen bonds between compound Q17 and two amino acid residues named GLU239 and LYS188, were uncovered by molecular docking and molecular dynamics simulation. The cell-based assays showed that compound Q17 could protect the SH-SY5Y human neuroblastoma cell line from okadaic acid (OA)-induced injury by targeting DYRK1A. More importantly, compound Q17 significantly improved cognitive dysfunction of 3 × Tg-AD mice, ameliorated pathological changes, and attenuated Tau hyperphosphorylation as well as Aß deposition. In summary, our computational modeling strategy is effective to identify novel chemotypes of DYRK1A inhibitors with great potential to treat AD, and the identified compound Q17 in this study is worthy of further study.

In Situ Oral Metabolism Analysis of Astringent Compounds in Tea by Paper Spray Mass Spectrometry, Electrospray Mass Spectrometry, Turbidimetry, and Sensory Evaluation.

The phenolic compounds (PCs) are the primary components responsible for the astringency of tea infusions, and this astringency is intricately linked to the in situ oral metabolism of PCs in saliva. Initially, a total of 54 PCs were identified in tea infusions by electrospray mass spectrometry (ESI-MS). Subsequently, an in vivo metabolism analysis of PCs during varying drinking times and oral locations was conducted by both paper spray mass spectrometry (PS-MS) and sensory evaluation. The metabolism of PCs within oral saliva was a prolonged process, the residual PCs were distributed across diverse oral regions after drinking tea infusion, and the higher residual PC content reflected the stronger astringency intensity. Furthermore, an in vitro metabolism analysis of PCs under varied reaction temperatures and durations was performed by ESI-MS and turbidimetry. As the reaction time extended, more PCs in tea was interacting with saliva. Moreover, the higher temperatures facilitated this interaction between PCs and saliva. Therefore, this investigation establishes a foundation for further elucidating the mechanisms underlying astringency formation.

Discovery and Flavor Characterization of High-Grade Markers in Baked Green Tea.

Green tea is a popular beverage around the world and possesses a unique flavor. The flavor qualities of green tea are closely related to its grade and this relationship has not yet been studied. Three baked green teas with similar flavor were studied, namely, Huangshan Maofeng, Taiping Houkui, and Shucheng Xiaolanhua. A total of 34 odor compounds were identified by solid phase microextraction (SPME) combined with two-dimensional comprehensive gas chromatography-olfactometry-mass spectrometry analysis (GC×GC-O-MS). The results of the clustering analysis showed that the content of D-limonene and linalool in the high-grade (Grade A) tea was much higher than the content in other grades, so they were identified as odor markers of Grade A baked green tea. The taste components of different grades of green tea infusion were analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS) and HPLC. A combination of clustering analysis, principal component analysis (PCA), and orthogonal partial least squares discrimination analysis (OPLS-DA) indicated that galloylglucose, digalloylglucose, trigalloyglucose, strictinin, and gallic acid could be used as taste markers of Grade A baked green tea. Therefore, the results in this paper reveal the substances responsible for the odor and taste markers of high-grade baked green tea.

FAPD: An Astringency Threshold and Astringency Type Prediction Database for Flavonoid Compounds Based on Machine Learning.

Astringency is a puckering or velvety sensation mainly derived from flavonoid compounds in food. The traditional experimental approach for astringent compound discovery was labor-intensive and cost-consuming, while machine learning (ML) can greatly accelerate this procedure. Herein, we propose the Flavonoid Astringency Prediction Database (FAPD) based on ML. First, the Molecular Fingerprint Similarities (MFSs) and thresholds of flavonoid compounds were hierarchically clustering analyzed. For the astringency threshold prediction, four regressions models (i.e., Gaussian Process Regression (GPR), Support Vector Regression (SVR), Random Forest (RF), and Gradient Boosted Decision Tree (GBDT)) were established, and the best model was RF which was interpreted by the SHapley Additive exPlanations (SHAP) approach. For the astringency type prediction, six classification models (i.e., RF, GBDT, Gaussian Naive Bayes (GNB), Support Vector Machine (SVM), k-Nearest Neighbor (kNN), and Stochastic Gradient Descent (SGD)) were established, and the best model was SGD. Furthermore, over 1200 natural flavonoid compounds were discovered and built into the customized FAPD. In FAPD, the astringency thresholds were achieved by RF; the astringency types were distinguished by SGD, and the real and predicted astringency types were verified by t-Distributed Stochastic Neighbor Embedding (t-SNE). Therefore, ML models can be used to predict the astringency threshold and astringency type of flavonoid compounds, which provides a new paradigm to research the molecular structure-flavor property relationship of food components.

Crystal Facet-Modulated WO3 Nanoplate Photoanode for Photoelectrochemical Glyoxal Semi-oxidation into Glyoxylic Acid.

Transforming glyoxal to value-added glyoxylic acid (GA) is highly desirable but challenging due to the uncontrollable over-oxidation. In this work, we report on a first demonstration of semi-oxidation of glyoxal with high selectivity (86.5%) and activity on WO3 nanoplate photoanode through the photoelectrochemical strategy. The optimization of reactivity was achieved via crystal facet regulation, showing a satisfactory GA production rate of 308.4 mmol m-2 h-2, 84.0% faradaic efficiency, and 4.3% total solar-to-glyoxylic acid efficiency on WO3 with enriched {200} facets at 1.6 V versus RHE. WO3 with a high {200} facet ratio exhibits more efficient electron-hole transfer kinetics, resulting in the facilitated formation of hydroxyl radicals (•OH) and glyoxal radicals. Meanwhile, the theoretical calculation results indicate that the high selectivity and activity come from the strong adsorption ability for glyoxal and the low reaction energy for glyoxal radical generation on the (200) facets of WO3. Moreover, the high energy demand toward oxalic acid production on WO3 leads to the exciting semi-oxidation process.

Development of Mo-Modified Pseudoboehmite Supported Ni Catalysts for Efficient Hydrogen Production from Formic Acid.

Formic acid (FA), as a safe and renewable liquid hydrogen storage material, has attracted extensive attention. In this paper, a series of Mo-modified pseudoboehmite supported Ni catalysts were developed and evaluated for efficient hydrogen production from formic acid. Pseudoboehmite (PB) as a catalyst carrier was used for the first time. Ni/PB and NiMo/PB possessed a mesostructure, and the pore size distribution was mainly concentrated between 2 and 20 nm. The oxygen vacancies caused by Mo enhanced Ni anchoring, thus inhibiting Ni sintering. Compared with Ni10/PB (7.62 nm), Ni10Mo1/PB had smaller Ni particles (5.08 nm). The Ni-O-Al solid solutions formed through the interaction of Ni with the PB improved the catalytic performance. Ni10Mo1/PB gave the highest conversion of 92.8% with a H2 selectivity of 98% at 300 °C, and the catalyst activity hardly decreased during the 50 h stability test. In short, Ni10Mo1/PB was a promising catalyst for hydrogen production from formic acid because of the oxygen vacancy anchoring effect as well as the formation of Ni-O-Al solid solutions which could effectively suppress the Ni sintering.

TiO2@PDA inorganic-organic core-shell skeleton supported Pd nanodots for enhanced electrocatalytic hydrodechlorination.

The development of catalysts with high atom utilization and activity is the biggest challenge for electrocatalytic hydrodechlorination (EHDC) technology. Herein, a design strategy of TiO2@PDA inorganic-organic core-shell skeleton for loading lower dosage of noble palladium (Pd) with robust activity is reported. The self-supported TiO2@PDA nanorod arrays provides exposed surface area for anchoring Pd and PDA as interlayer controls the Pd nucleation to form nanodots with high dispersion, realizing high atom utilization. Moreover, the strong interaction between PDA and Pd realizes the coexistence of electron-rich and deficient Pd species with suitable proportion, which facilitate the H* formation and the C-Cl bond activation, respectively, resulting in the promoted activity. The optimal TiO2@PDA/Pd electrode exhibits a low dosage of Pd (0.093 mg cm-2) and excellent activity for 4-chlorophenol reduction with a mass activity (MA) of 23.96 min-1g-1, which is 3.31 times as high as that of TiO2/Pd. The design scheme with inorganic-organic core-shell skeleton as support is benefit for developing highly efficient and lower price elctrocatalysts for EHDC.

Differences of characteristic aroma compounds in Rougui tea leaves with different roasting temperatures analyzed by switchable GC-O-MS and GC × GC-O-MS and sensory evaluation.

Roasting, an important process to refine Wuyi Rock tea, could impart different types of aroma to the final products. This study focuses on the differences in aroma characteristics among three kinds of refined teas, named light fire (LF), moderate fire (MF), and high fire (HF). A combination of solid phase microextraction (SPME) and a switchable system between GC-O-MS and GC × GC-O-MS was utilized to identify the odorants. In total, 97 aroma-active compounds could be smelled at the sniffing port, comprising alcohols, aldehydes, ketones, esters, heterocycles, and terpenes. However, only 52 obtained r-OAV >1. Significant differences were uncovered by the application of principal component analysis (PCA) and partial least squares regression (PLSR). Thereby, MF and HF had a more similar aroma profile, while in LF samples, alcohols, aliphatic aldehydes and some ketones were responsible for the aroma profile, such as (E,E)-2,4-hexadienal, octanal, hexanal, (E,Z)-2,6-nonadienal, (E)-ß-ionone, 3-octen-2-one etc. Strecker aldehydes had a great impact on the aroma of MF, including 2-methylpropanal, 2-methylbutanal, 3-methylbutanal etc. Some N-heterocyclic compounds also affected the overall aroma, for instance, 6-methyl-2-ethylpyrazine. In HF, the majority of aroma compounds increased with increasing roasting temperature, especially N-heterocyclic compounds as well as furfural and 5-methyl-2-furancarboxaldehyde, which are all closely related to the Maillard reaction. Besides, 5-methyl-2-(1-methylethenyl)-4-hexen-1-ol, trans-linalooloxide and 2-nonanone also remarkably influenced the aroma of HF. In addition, it was supposed that most amino acids that participated in the Maillard reaction during roasting were decomposed from the compounds that combined with tea polyphenols and amino acids.

Rapid visualized characterization of phenolic taste compounds in tea extract by high-performance thin-layer chromatography coupled to desorption electrospray ionization mass spectrometry.

Phenolic compounds are the important taste source of tea infusion. In this paper, the phenolic compounds in tea extracts were separated by high-performance thin-layer chromatography (HPTLC), and then in-situ determined by desorption electrospray ionization mass spectrometry (DESI-MS). Total 44 phenolic compounds in tea extracts were accurately confirmed by NIST library as well as reference substances. The clustering results of heat-map can better reflect the differences of phenolic compounds in different categories and subcategories of teas. Besides, the contents of hydrolyzable tannins, including galloylglucose, digalloylglucose, trigalloyglucose and strictinin, were positively correlated with the grades of green tea. The method validation and quantification results of exemplified five phenolic compounds in teas were also obtained, and LODs, LOQs and recoveries were ranging between 1.5-15.9 µg/mL, 5.1-53.1 µg/mL, and 79%-117.6%, respectively. Moreover, HPTLC-DESI-MS can save tenfold analytical time compared to HPLC-MS. Therefore, HPTLC-DESI-MS was a rapid, efficient characterization method of phenolic compounds in tea extracts.

Structural identification of sour compounds in wine and tea by ambient ionization mass spectrometry according to characteristic product ion and neutral loss.

Sourness is an important food taste for human. A rapid, accurate method was used to generalize the structure similarity and diversity of sour compounds. Based on the product ion and neutral loss of sour compounds, ambient ionization techniques combined with quadrupole-Orbitrap mass spectrometry (AI-Q-Orbitrap) was employed. According to the behavior of sour compounds in the process of high collision dissociation (HCD) of MS/MS, three fragmentation pathway schemes were proposed: (1) charge-driven fragmentation and CO2 loss, (2) six-membered ring rearrangement and Cα-Cß cleavage, and (3) elimination rearrangement and H2O, CO2 and CO loss in succession. Besides, structure information about characteristic product ions and characteristic neutral losses was summarized. Finally, multi-class sour compounds including monoacids, diacids, polyacids and phenolic acids in wine and tea were identified and compared. Therefore, sour compounds and their structure information can be determined by AI-MS based on characteristic product ion and neutral loss.

Monomethylfumarate protects against ovariectomy-related changes in body composition.

Osteoporosis, low bone mass that increases fracture susceptibility, affects approximately 75 million individuals in the United States, Europe and Japan, with the number of osteoporotic fractures expected to increase by more than 3-fold over the next 50 years. Bone mass declines with age, although the mechanisms for this decrease are unclear. Aging enhances production of reactive oxygen species, which can affect bone formation and breakdown. The multiple sclerosis drug Tecfidera contains dimethylfumarate, which is rapidly metabolized to monomethylfumarate (MMF); MMF is thought to function through nuclear factor erythroid-derived-2-like-2 (Nrf2), a transcription factor activated by oxidative stress which induces the expression of endogenous anti-oxidant systems. We hypothesized that MMF-elicited increases in anti-oxidants would inhibit osteopenia induced by ovariectomy, as a model of aging-related osteoporosis and high oxidative stress. We demonstrated that MMF activated Nrf2 and induced anti-oxidant Nrf2 target gene expression in bone marrow-derived mesenchymal stem cells. Sham-operated or ovariectomized adult female mice were fed chow with or without MMF and various parameters monitored. Ovariectomy produced the expected effects, decreasing bone mineral density and increasing body weight, fat mass, bone marrow adiposity and serum receptor activator of nuclear factor-kappa-B ligand (RANKL) levels. MMF decreased fat but not lean mass. MMF improved trabecular bone microarchitecture after adjustment for body weight, although the unadjusted data showed few differences; MMF also tended to increase adjusted cortical bone and to reduce bone marrow adiposity and serum RANKL levels. Because these results suggest the possibility that MMF might be beneficial for bone, further investigation seems warranted.

Quantitation of isoprenoids for natural rubber biosynthesis in natural rubber latex by liquid chromatography with tandem mass spectrometry.

The natural rubber molecule is one of the end products of isoprenoids metabolism in the plant. Dimethylallyl diphosphate (DMAPP) and farnesyl pyrophosphate (FPP) are two typical isoprenoids which control the rate of biosynthesis and the molecular weight of natural rubber. A rapid, nonradioactive method for quantitation of DMAPP and FPP in natural rubber latex by liquid chromatography tandem with mass spectrometry (LC-MS/MS) was reported. DMAPP and FPP were determined in the multiple reaction monitoring mode(MRM)followed by separation with a silica-based C18 column. The external standard quantitative method was established, and the results showed limits of quantitation (LOQs) were 28 ng/ml and 33 ng/ml for DMAPP and FPP, respectively. The concentrations were detected 70-96 ng/ml and 242-375 ng/ml for these two isoprenoids in natural rubber latex. Recoveries of the method were in the range 81-93%. Daytime comparison experiments found that FPP had better stability than DMAPP.

Applications of DART-MS for food quality and safety assurance in food supply chain.

Direct analysis in real time (DART) represents a new generation of ion source which is used for rapid ionization of small molecules under ambient conditions. The combination of DART and various mass spectrometers allows analyzing multiple food samples with simple or no sample treatment, or in conjunction with prevailing protocolized sample preparation methods. Abundant applications by DART-MS have been reviewed in this paper. The DART-MS strategy applied to food supply chain (FSC), including production, processing, and storage and transportation, provides a comprehensive solution to various food components, contaminants, authenticity, and traceability. Additionally, typical applications available in food analysis by other ambient ionization mass spectrometers were summarized, and fundamentals mainly including mechanisms, devices, and parameters were discussed as well. © 2015 Wiley Periodicals, Inc. Mass Spec Rev. 36:161-187, 2017.

Rapid screening and quantification of residual pesticides and illegal adulterants in red wine by direct analysis in real time mass spectrometry.

A rapid method to screen and quantify multi-class analytic targets in red wine has been developed by direct analysis in real time (DART) coupled with triple quadruple tandem mass spectrometry (QqQ-MS). A modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) procedure was used for increasing analytical speed and reducing matrix effect, and the multiple reaction monitoring (MRM) in DART-MS/MS ensured accurate analysis. One bottle of wine containing 50 pesticides and 12 adulterants, i.e., preservatives, antioxidant, sweeteners, and azo dyes, could be totally determined less than 12min. This method exhibited proper linearity (R2≥0.99) in the range of 1-1000ng/mL for pesticides and 10-5000ng/mL for adulterants. The limits of detection (LODs) were obtained in a 0.5-50ng/mL range for pesticides and 5-50ng/mL range for adulterants, and the limits of quantification (LOQs) were in a 1-100ng/mL range for pesticides and 10-250ng/mL range for adulterants. Three spiked levels for each analyte in wine were evaluated, and the recoveries were in a scope of 75-120%. The results demonstrated DART-MS/MS was a rapid and simple method, and could be applied to rapid analyze residual pesticides and illegal adulterants in a large quantities of red wine.

Simultaneous qualitation and quantitation of natural trans-1,4-polyisoprene from Eucommia ulmoides Oliver by gel permeation chromatography (GPC).

Natural trans-1,4-polyisoprene (TPI) as a functional biomaterial has aroused great interest for rubber industrial product use. Here, we proposed a method that enables simultaneous analysis of the content and molecular-weight distribution (MWD) of natural TPI by gel permeation chromatography (GPC). The natural TPIs were collected from leaves, fruit coatings and bark of Eucommia ulmoides Oliver (E. ulmoides) through toluene extraction followed by ethanol purification. The results of TPI contents from leaves and fruit coatings were shown ca. 3.5% and 13.8%, respectively. Accordingly, limits of detection (LODs) of TPI were 0.58mg/mL from leaves and 0.47mg/mL from fruit coatings. The MWDs of TPI demonstrated a bimodal distribution from leaves, a unimodal distribution from bark, and a unimodal distribution with a tiny peak shoulder from fruit coatings. In real-life E. ulmoides analysis, the results from three independent methods (GPC, gravimetric method, and infrared spectroscopy) were obtained with good consistency.

Determination of Dicyandiamide in Powdered Milk Using Direct Analysis in Real Time Quadrupole Time-of-Flight Tandem Mass Spectrometry.

The direct analysis in real time (DART) ionization source coupled with quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS) system has the capability to desorb analytes directly from samples without sample cleanup or chromatographic separation. In this work, a method based on DART/Q-TOF MS/MS has been developed for rapid identification of dicyandiamide (DCD) present in powdered milk. Simple sample extraction procedure employing acetonitrile-water (80:20, v/v) mixture was followed by direct, high-throughput determination of sample extracts spread on a steel mesh of the transmission module by mass spectrometry under ambient conditions. The method has been evaluated for both qualitative and quantitative analysis of DCD in powdered milk. Variables including experimental apparatus, DART gas heater temperature, sample presentation speed, and vacuum pressure were investigated. The quantitative method was validated with respect to linearity, sensitivity, repeatability, precision, and accuracy by using external standards. After optimization of these parameters, a limit of detection (LOD) of 100 µg kg(-1) was obtained for DCD with a linear working range from 100 to 10000 µg kg(-1) and a satisfactory correlation coefficient (R(2)) of 0.9997. Good recovery (80.08%-106.47%) and repeatability (RSD = 3.0%-5.4%) were achieved for DCD. The DART/Q-TOF MS/MS-based method provides a rapid, efficient, and powerful scheme to analyze DCD in powdered milk with limited sample preparation, thus reducing time and complexity of quality control.