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Interest in galactomannans and its derivatives as a functional health supplement is growing based on physicochemical properties. In this work, the optimized conditions of sulfated tara gum (STG) with a maximum DS of 0.66 by box-behnken design (BBD) were obtained as following: ratio of chlorosulfonic acid/pyridine 3:1, reaction time 4 h and reaction temperature 40 °C. The structure features of STG such as the degree of substitution (DS), substitution position, weight average molar mass (MW), monosaccharide components and chain conformation were investigated. Decreasing of MW, the increasing of Z-average radius of gyration (〈S2〉Z1/2) and specific volume for gyration (SVg) were obtained by SEC-MALLS. In addition, the structural properties of four sulfated galactomannans were comparatively investigated and analyzed based on our earlier reports of sulfated fenugreek gum, guar gum and locust bean gum. A conclusion was drown that higher galactose branch could enhance steric hindrance, which was inferred as one of the significant factors for the derivatization efficiency, thus affecting the DS, MW and conformational transition of sulfated galactomannans. This study will provide valuable information for further research on the comparison of bioactivities and medical application of galactomannans family.

Precise positioning using smartphones has been a topic of interest especially after Google decided to provide raw GNSS measurement through their Android platform. Currently, the greatest limitations in precise positioning with smartphone Global Navigation Satellite System (GNSS) sensors are the quality and availability of satellite-to-smartphone ranging measurements. Many papers have assessed the quality of GNSS pseudorange and carrier-phase measurements in various environments. In addition, there is growing research in the inclusion of a priori information to model signal blockage, multipath, etc. In this contribution, numerical estimation of actual range errors in smartphone GNSS precise positioning in realistic environments is performed using a geodetic receiver as a reference. The range errors are analyzed under various environments and by placing smartphones on car dashboards and roofs. The distribution of range errors and their correlation to prefit residuals is studied in detail. In addition, a comparison of range errors between different constellations is provided, aiming to provide insight into the quantitative understanding of measurement behavior. This information can be used to further improve measurement quality control, and optimize stochastic modeling and position estimation processes.

Global warming is known to affect the growth, development and reproduction of insects. In this study, the larvae developmental process and endogenous hormone levels under high temperature (36 °C) stress were investigated in the lepidopteran model insect Bombyx mori (B. mori). After high temperature treatment, the duration of 5th instar larvae was shortened by 28 ± 2 h, the content of 20-hydroxyecdysone(20E) in hemolymph was significantly increased, and the transcription levels of the 20E response genes E93, Br–C, USP and E75 were up-regulated by 1.35-, 1.25-, 1.28-, and 1.27-fold, respectively. High temperature treatment also elevated the phosphorylation level of Akt and activated the downstream BmCncC/keap1 pathway, and the transcription levels of the 20E synthesis-related genes cyp302a1, cyp306a1, cyp314a1 and cyp315a1 were up-regulated by 1.12-, 1.51-, 2.17- and 1.23-fold, respectively. The transcription levels of cyp302a1 and cyp306a1 were significantly decreased in BmN cells after treatment with the double stranded RNA of BmCncC (dsBmCncC), whereas their transcription levels were significantly increased (2.15- and 1.31-fold, respectively) after treatment with the CncC agonist Curcumin. These results demonstrated that high temperature treatment promoted the metamorphosis and the BmCncC/keap1 pathway played a role in the metamorphosis of B. mori. Our results provided clues for understanding the CncC/keap1 pathway-mediated regulation of metamorphosis of Lepidopteran insects.

The growth and proliferation of most cancer cells involve the excessive uptake of glucose mediated by glucose transporters. An effective strategy for cancer therapy has been to inhibit the GLUTs that are usually overexpressed in a variety of tumor cells. 2-NBDG is a GLUT1 substrate that can be used as a probe for GLUT1 inhibitors. An accurate and simple assay for 2-NBDG in a HEK293T cell model overexpressing GLUT1 was developed using liquid chromatography-tandem mass spectrometry. Chromatographic separation was achieved using a Xbridge® Amide column (3.5 μm, 2.1 mm × 150 mm, Waters) with acetonitrile-water containing 2 μM ammonium acetate (80:20, v/v) at a flow rate of 0.25 mL/min. Mass detection was conducted in the parallel reaction monitoring (PRM) mode. The calibration curve for 2-NBDG showed good linearity in the concentration range of 5–500 ng/mL with satisfactory precision, a relative standard deviation ranging from 2.92 to 9.59% and accuracy with a relative error ranging from −13.14 to 7.34%. This method was successfully applied to quantify the uptake of GLUT1-mediated 2-NBDG, and the results clearly indicated inhibition of GLUT1 by WZB117 and quercetin (two potent glucose transporter inhibitors) in the GLUT1-HEK293T cell model. This study provides a convenient and accurate method for high-throughput screening of selective and promising GLUT1 inhibitors.

Silk gland is a major organ of Bombyx mori for the synthesis and secretion of silk protein. Phoxim exposure can be toxic to B. mori and causes a decrease of fibroin synthesis, finally affecting the silk production in industry. To study the mechanism of metabolism and detoxification of silk gland under phoxim exposure, we measured the residual quantity of phoxim in silk gland and hemolymph after phoxim exposure, and the detoxifying enzymes-related genes and enzyme activity were also investigated. Results indicated that the residual amount of phoxim existed up to 24 h in silk gland compared with that in hemolymph, suggesting that phoxim can accumulate in the silk glands within a certain time course. The transcriptional levels of PI3K/Akt genes, including Akt, Tor1, p70s6k and 4e-bp, were up-regulated by 6.919, 1.358, 10.766 and 7.708-fold, respectively. The expression of two downstream genes (CncC and Keap1) was up-regulated by 1.939 and 3.373-fold, respectively. In addition, the transcriptional levels of detoxification-related genes including CYP6AB, CYP306A, CarE2, GST1 and GSTd1 were up-regulated by 1.731, 1.221, 1.366, 1.376 and 6.591-fold, respectively. The enzymatic activity of CYP450, CarE and GST were increased over time. These results provided possible insights into the injury of silk gland and the transcriptional response to detoxifying enzymes-related genes in silkworm after phoxim exposure.

Folate metabolism pathway is mainly associated with syntheses of nucleic acids and proteins as well as DNA methylation and repair. Cytoplasmic C1 tetrahydrofolate (C1-THF) synthase is a central enzyme in folate metabolism and plays an important role in C1-THF conversion. The full-length sequence of silkworm (Bombyx mori) cytoplasmic C1-THF synthase (BmC1-THF synthase) gene including an open reading frame (ORF) of 932 amino acid residues was cloned. Phylogenetic analysis indicated that BmC1-THF synthase shares 84% and 81% identity with C1-THF synthase of Papilio Xuthus and Danaus plexippus, respectively. Conserved sequence analysis showed that the 330 to 710 amino acid residues of BmC1-THF synthase were highly conserved among various organisms. Recombinant BmC1-THF synthase protein with a molecular weight of about 105.5 kDa was expressed in E. coli cells and identified using SDS-PAGE and western blot analysis. The expression of BmC1-THF synthase gene in eight silkworm tissues revealed that BmC1-THF synthase gene is widely expressed in various tissues of silkworm and highly expressed in the fat body. The expression characteristics of BmC1-THF synthase gene in response to oxidative stress were conducted under phoxim and high temperature (30 °C) exposure. The results showed that the transcriptional level of BmC1-THF synthase gene was upregulated by 3.435-fold and 6.845-fold after phoxim and high temperature (30 °C) exposure, respectively, indicating that BmC1-THF synthase gene may play a significant role in response of silkworm to oxidative stresses.

Various nanoparticles, such as silver nanoparticles (AgNPs) and titanium nanoparticles (TiO2 NPs) are increasingly used in industrial processes. Because they are released into the environment, research into their influence on the biosphere is necessary. Among its other effects, dietary TiO2 NPs promotes silk protein synthesis in silkworms, which prompted our hypothesis that TiO2 NPs influence protein kinase B (Akt)/Target of rapamycin (Tor) signaling pathway (Akt/Tor) signaling in their silk glands. The Akt/Tor signaling pathway is a principle connector integrating cellular reactions to growth factors, metabolites, nutrients, protein synthesis, and stress. We tested our hypothesis by determining the influence of dietary TiO2 NPs (for 72 h) and, separately, of two Akt/Tor pathway inhibitors (LY294002 and rapamycin) on expression of Akt/Tor signaling pathway genes and proteins in the silk glands. TiO2 NPs treatments led to increased accumulation of mRNAs for Akt, Tor1 and Tor2 by 1.6-, 12.1-, and 4.8-fold. Dietary inhibitors led to 2.6- to 4-fold increases in mRNAs encoding Akt and substantial decreases in mRNAs encoding Tor1 and Tor2. Western blot analysis showed that dietary TiO2 NPs increased the phosphorylation of Akt and its downstream proteins. LY294002 treatments led to inhibition of Akt phosphorylation and its downstream proteins and rapamycin treatments similarly inhibited the phosphorylation of Tor-linked downstream proteins. These findings support our hypothesis that TiO2 NPs influence Akt/Tor signaling in silk glands. The significance of this work is identification of specific sites of TiO2 NPs actions.

Bombyx mori is an economic insect of the Lepidoptera. Its posterior silk gland (PSG) is an important organ for fibroin synthesis. In order to study the occurrence of apoptosis in PSG and the role of PI3K/Akt signaling pathway during spinning period, changes in morphology of silk gland, expressions of fibroin components Fib-H, Fib-L and P25 and Akt, TOR2, P70S6K and S6 in PI3K/Akt pathway, expressions of apoptosis related genes caspase-3, caspase-9 and activity of caspase-3 were explored. The results showed that the morphology of silk gland dramatically degenerated; transcription of Fib-H, Fib-L, and P25 gradually declined with time; and Fib-L protein level reduced by 0.6-fold at 72 h. Moreover, the transcription levels of Akt, TOR2, P70S6K, and S6 also decreased by 0.3-, 0.8-, 0.7-, and 0.1-fold, respectively, indicating that the downregulation of PI3K/Akt signaling pathway could lead to reduction in fibroin synthesis. In addition, the transcription levels of caspase-3 and caspase-9 increased by 1.3- and 3.6-fold, respectively, and the enzyme activity of caspase-3 grew at a maximum of 1.6-fold. The results showed the occurrence of apoptosis in PSG during spinning period. In conclusion, the present study indicated that both the decline in fibroin components and the increase in apoptosis-related genes were regulated by PI3K/Akt signaling pathway during spinning period, which shed new light on the functions of PI3K/Akt signaling pathway.

Aminoacyl-tRNA synthetases are the key enzymes for protein synthesis. Glycine, alanine, serine and tyrosine are the major amino acids composing fibroin of silkworm. Among them, the genes of alanyl-tRNA synthetase (AlaRS) and glycyl-tRNA synthetase (GlyRS) have been cloned. In this study, the seryl-tRNA synthetase (SerRS) and tyrosyl-tRNA synthetase (TyrRS) genes from silkworm were cloned. Their full length are 1709 bp and 1868 bp and contain open reading frame (ORF) of 1485 bp and 1575 bp, respectively. RT-PCR examination showed that the transcription levels of SerRS, TyrRS, AlaRS and GlyRS are significantly higher in silk gland than in other tissues. In addition, their transcription levels are much higher in middle and posterior silk gland than in anterior silk gland. Moreover, treatment of silkworms with phoxim, an inhibitor of silk protein synthesis, but not TiO2 NP, an enhancer of silk protein synthesis, significantly reduced the transcription levels of aaRS and content of free amino acids in posterior silk gland, therefore affecting silk protein synthesis, which may be the mechanism of phoxim-silking disorders. Furthermore, low concentration of TiO2 NPs showed no effect on the transcription of aaRS and content of free amino acids, suggesting that TiO2 NPs promotes silk protein synthesis possibly by increasing the activity of fibroin synthase in silkworm.

Insect fat body is an important intermediate metabolic organ that plays an important role in protein metabolism and detoxification. In order to study the effects of TiO2 NPs and phoxim on fat body protein synthesis through MAPK and PI3K/Akt signaling pathways in silkworms, we determined the effects of TiO2 NPs and phoxim, alone and in combination, on fat body protein content of silkworms, analyzed the gene expression profile of the fat body, and verified the expression of characteristic genes. We found that TiO2 NPs and phoxim alone increased the total protein content of the fat body, and up-regulated MAPK and PI3K/Akt signaling pathway genes. TiO2 NPs up-regulated the expression of two growth and development-related genes-insulin-like peptide and neuropeptide receptor B-by 5.17 and 3.89-fold, respectively. Phoxim up-regulated the expression of detoxification genes-P450, GST, and CarE2. Pretreatment with TiO2 NPs could reduce phoxim-increased total protein content and up-regulated MAPK and PI3K/Akt signaling pathway genes and detoxification genes; the activities of detoxification enzymes were consistent with the gene expression pattern. Our results showed that MAPK and PI3K/Akt signaling pathways both regulate fat body protein synthesis in silkworms, but the target proteins induced to express were different under different inducing factors. Our finding may provide a reference for investigating the mechanism of protein synthesis regulation through MAPK and PI3K/Akt signaling pathways.