Stability studies and structure identification of anthocyanins from Tradescantia pallida

Anthocyanins are an important type of natural red pigments for use in replacing the artificial red colorants in foods. The major problem with anthocyanins is their instability. The search for more stable anthocyanins, especially at near neutral pH, was the motivation for this research. The anthocyanins from the leaves and stems of Tradescantia pallida were found to be more stable than conventional anthocyanins of grapes and blackberries at near neutral pH, such as pH 5.5, in a model beverage storage study of more than 54 weeks. The high stability of this type of anthocyanin may eventually expand the application of anthocyanin from high acid to low acid foods. Furthermore, the stability of T. pallida anthocyanins(TPA) was significantly reduced by tannic acid, ascorbic acid and their interaction in a 2-factor 3-level factorial experiment, in contrast to the enhancement by tannic acid, of the color and stability of the conventional anthocyanins from grapes and blackberries. Because of the significant stability of TPA, understanding of their chemical structure became necessary. A new approach of identifying anthocyanin structure was developed, which includes both qualitatively identifying the deacylated anthocyanins, with comparison to the known deacylated anthocyanins of Zebrina pendula through HPLC and TLC, the sugars with TLC, and the acyl groups with TLC and HPLC, and quantitatively measuring the ratio of sugar to the aglycone with the dinitrosalicylic acid method and the acyl/aglycone ratio with HPLC. The pigments of T. pallida were found to consist of mainly two anthocyanins, one in much higher abundance than the other. The major one was identified as cyanidin-3,7,-diglucosylferuloyl-3\sp\prime-glycosylferuloylglucoside, and the minor one cyanidin-3,7,3\sp\prime-triglycosylferulic acid. This study indicated the possibility of applying this chemical approach of anthocyanin structure identification especially to those anthocyanins whose acyl groups were qualitatively identified but not quantitatively measured

Water Pricing Policy in Tarim Basin of China

China ranks the thirteenth among countries with serious water shortage problems in the world. The average amount of water owned per person is only 2400 m3/year, which is about 1/4 of the world average. But unfortunately, the efficiency of water use, especially in agriculture in some arid areas, is very low, only 20% - 30% in some areas. The main reason is that water prices are too low to protect the water resources. In this paper, the Tarim Basin of southern Xinjiang is selected to study the water supply costs and farmer's tolerance of water expenses based on a great amount of data collected in the four prefectures in the Tarim Basin. Then, three steps are suggested for water pricing reform in the Tarim Basin. Finally, several possible water pricing patterns are presented, such as water coupons, seasonal floating prices, and water price counting in kind but paying in currency. The conclusion is that the present water price system should be reformed and the water price can be increased to some extent for agricultural use even in Xinjiang, a developing area in China

Effect of cerebellar transcranial magnetic stimulation with double-cone coil on dysphagia after subacute infratentorial stroke: A randomized, single-blinded, controlled trial

Background: A 10-Hz cerebellar repetitive transcranial magnetic stimulation (rTMS) could increase corticobulbar tract excitability in healthy individuals. However, its clinical efficacy for poststroke dysphagia (PSD) remains unclear. Objective: To investigate the effectiveness of 10-Hz cerebellar rTMS in PSD patients with infratentorial stroke (IS). Methods: In this single-blinded, randomized controlled trial, 42 PSD patients with subacute IS were allocated to three groups: bilateral cerebellar rTMS (biCRB-rTMS), unilateral cerebellar rTMS (uniCRB-rTMS), or sham-rTMS. The stimulation parameters were 5 trains of 50 stimuli at 10 Hz with an interval of 10 s at 90% of the thenar resting motor threshold (RMT). The Functional Oral Intake Scale (FOIS) was assessed at T0 (baseline), T1 (day 0 after intervention), and T2 (day 14 after intervention), whereas the Dysphagia Outcome and Severity Scale (DOSS), Penetration Aspiration Scale (PAS), and neurophysiological parameters were evaluated at T0 and T1. Results: Significant time and intervention interaction effects were observed for the FOIS score (F = 3.045, p = 0.022). The changes in the FOIS scores at T1 and T2 were both significantly higher in the biCRB-rTMS group than in the sham-rTMS group (p < 0.05). The uniCRB-rTMS and biCRB-rTMS groups demonstrated greater changes in the DOSS and PAS at T1, compared with the sham-rTMS group (p < 0.05). Bilateral corticobulbar tract excitability partly increased in the biCRB-rTMS and uniCRB-rTMS groups at T1, compared with T0. The percent changes in corticobulbar tract excitability parameters at T1 showed no difference among three groups. Conclusions: A 10-Hz bilateral cerebellar rTMS is a promising, noninvasive treatment for subacute infratentorial PSD

CX-MS Datasets for "Comprehensive Structure and Functional Adaptations of the Yeast Nuclear Pore Complex"

Data Files Description: NPC_XL_Identification_Inter_Crosslinked.csv: Inter-protein cross-links identified by pLink 2; NPC_XL_spectra.mgf: MS2 spectra data for the identified cross-links; NPC_XL_proteins.fasta : Protein sequences used for search.This repository contains chemical cross-linking mass spectrometry data of affinity-purified Yeast nuclear pore complexes.[Sample Processing] NPCs were immuno-purified from Mlp1 tagged S. cerevisiae strains (Kim et al., 2018). After native elution, 1.0 mM disuccinimidyl suberate (DSS) was added and the sample was incubated at 25ºC for 40 min with shaking (1,200 rpm). The reaction was quenched by adding a final concentration of 50 mM freshly prepared ammonium bicarbonate and incubating for 20 min with shaking (1,200 rpm) at 25ºC. The sample (50 µg) was then concentrated and denatured at 98ºC for 5 min in a solubilization buffer (10% solution of 1-dodecyl-3-methylimidazolium chloride (C12-mim-Cl) in 50 mM ammonium bicarbonate, pH 8.0, 100 mM DTT). After denaturation, the sample was centrifuged at 21,130 g for 10 min and the supernatant was transferred to a 100 kDa MWCO ultrafiltration unit (MRCF0R100, Microcon). The sample was quickly spun at 1,000 g for 2 min and washed twice with 50 mM ammonium bicarbonate. After alkylation (50 mM iodoacetamide), the cross-linked NPC in-filter was digested by trypsin and lysC O/N at 37ºC. After proteolysis, the sample was recovered by centrifugation and peptides were fractionated into 10-12 fractions by using a stage tip self-packed with basic C18 resins (Dr. Masch GmbH). Fractionated samples were pooled prior to LC/MS analysis. Desalted cross-link peptides were dissolved in the sample loading buffer (5% Methanol, 0.2% FA), separated with an automated nanoLC device (nLC1200, Thermo Fisher), and analyzed by an Orbitrap Q Exactive HFX (Pharma mode) mass spectrometer (Thermo Fisher) as previously described (Xiang et al., 2020; Xiang et al., 2021). Briefly, peptides were loaded onto an analytical column (C18, 1.6 μm particle size, 100 Å pore size, 75 μm × 25 cm; IonOpticks) and eluted using a 120-min liquid chromatography gradient. The flow rate was approximately 300 nl/min. The spray voltage was 1.7 kV. The QE HF-X instrument was operated in the data-dependent mode, where the top 10 most abundant ions (mass range 380 – 2,000, charge state 4 - 8) were fragmented by high-energy collisional dissociation (HCD). The target resolution was 120,000 for MS and 15,000 for tandem MS (MS/MS) analyses. The quadrupole isolation window was 1.8 Th; the maximum injection time for MS/MS was set at 200 ms.[Data processing] The raw data were searched with pLink2 (Chen et al., 2019b). An initial MS1 search window of 5 Da was allowed to cover all isotopic peaks of the cross-linked peptides. The data were automatically filtered using a mass accuracy of MS1 ≤ 10 ppm (parts per million) and MS2 ≤ 20 ppm of the theoretical monoisotopic (A0) and other isotopic masses (A+1, A+2, A+3, and A+4) as specified in the software. Other search parameters included cysteine carbamidomethyl as a fixed modification and methionine oxidation as a variable modification. A maximum of two trypsin missed-cleavage sites was allowed. The initial search results were obtained using a default 5% false discovery rate (FDR) expected by the target-decoy search strategy. Spectra were manually verified to improve data quality (Kim et al., 2018; Shi et al., 2014). Cross-linking data were analyzed and plotted with CX-Circos (http://cx-circos.net).N

Comprehensive structure and functional adaptations of the yeast nuclear pore complex.

Nuclear pore complexes (NPCs) mediate the nucleocytoplasmic transport of macromolecules. Here we provide a structure of the isolated yeast NPC in which the inner ring is resolved by cryo-EM at sub-nanometer resolution to show how flexible connectors tie together different structural and functional layers. These connectors may be targets for phosphorylation and regulated disassembly in cells with an open mitosis. Moreover, some nucleoporin pairs and transport factors have similar interaction motifs, which suggests an evolutionary and mechanistic link between assembly and transport. We provide evidence for three major NPC variants that may&nbsp;foreshadow functional specializations at the nuclear periphery. Cryo-electron tomography extended these studies, providing a model of the in situ NPC with a radially expanded inner ring. Our comprehensive model reveals features of the nuclear basket and central transporter, suggests a role for the lumenal Pom152 ring in restricting dilation, and highlights structural plasticity that may be required for transport