Search

Abstract With the proceeding of global warming and water eutrophication, the phenomenon of green tide has garnered significant societal interest. Consequently, researchers had increasingly focused on the potential applications of green algae biomass, particularly its polysaccharides. The polysaccharide serves as the primary active constituent of green algae and has demonstrated numerous advantageous biological activities, including antioxidant, antiviral, anticoagulant, hypolipidemic and immuno-modulatory activities. The favorable bioavailability and solubility of green algae oligosaccharides are attributed to their low molecular weight. So there has been a growing interest in researching green algae polysaccharides and oligosaccharides for the utilization of marine biological resources. This review summarized the extraction, purification, chemical structure, composition, biological activity, and potential applications prospect of polysaccharides and oligosaccharides derived from green algae. The review could be helpful for expanding the applications of polysaccharides and oligosaccharides of green algae. Graphical Abstract

An improved method was developed for the detection of 16 polycyclic aromatic hydrocarbons (PAHs) in mainstream cigarette smoke during combustion. The methodological approach involved matrix matching standard solution preparation, direct injection of the extraction solution, and measurement of standard linearity curves to quantitatively analyze the release of target compounds. Considering the diverse physical and chemical properties as well as distribution patterns of the 16 PAHs in cigarette smoke, the impact of room temperature and high-temperature ion sources on the response of target compounds was investigated. Additionally, the preparation methods of matrix solvents and the matrix effects on the quantitative results of target compounds were also explored. Furthermore, a systematic study was conducted on the smoke release distribution of commercially available cigarettes with different circumferences. The results demonstrated that direct injection of sample extract eliminates the need for sample purification, concentration, and other processing steps employed in current detection methods. This simplifies sample pretreatment procedures while reducing organic solvent usage and consumables consumption. Moreover, it resolves issues related to low operational efficiency when dealing with large quantities of samples. The use of matrix solvent to prepare standard solution effectively reduces the influence of matrix effect, and solves the technical problem of low recovery of target substance in flue gas caused by no purification treatment when using gas chromatography-tandem mass spectrometry (GC-MS/MS) to analyze 16 kinds of PAHs. Under the normal working parameters of the instrument, the electron impact ion source (EI) was used under high temperature to improve the ionization efficiency of the targets in complex matrix and to ensure the accurate analysis of targets with low concentrations. The correlation coefficients (R2) of the calibration curves for 16 PAHs are 0.996 4-0.999 8, the limits of dectection and the limits of quantification are 0.29-1.48, 0.95-4.94 ng/cigarette, respectively. Under different concentration levels, the average recovery is 87.71%-123.26%, and the precision is within 1.58%-9.45%. By analyzing the reference cigarette 3R4F, there is no significant difference between this method and those reported in the literature. The total amount of PAHs released from 58 brands of cigarette samples are 742.76-1 505.99 ng/cigarette. At the same time, with the decrease of tar content in cigarette, the release of PAHs will be reduced accordingly. This method is simple, practical, stable, and accurate, providing more technical means for the detection of trace PAHs in cigarette smoke.

Abstract Background Although GqPCR activation often leads to cell survival by activating the PI3K/AKT pathway, it was previously shown that in several cell types AKT activity is reduced and leads to JNK activation and apoptosis. The mechanism of AKT inactivation in these cells involves an IGBP1-coupled PP2Ac switch that induces the dephosphorylation and inactivation of both PI3K and AKT. However, the machinery involved in the initiation of PP2A switch is not known. Methods We used phospho-mass spectrometry to identify the phosphorylation site of PP2Ac, and raised specific antibodies to follow the regulation of this phosphorylation. Other phosphorylations were monitored by commercial antibodies. In addition, we used coimmunoprecipitation and proximity ligation assays to follow protein–protein interactions. Apoptosis was detected by a TUNEL assay as well as PARP1 cleavage using SDS-PAGE and Western blotting. Results We identified Ser24 as a phosphorylation site in PP2Ac. The phosphorylation is mediated mainly by classical PKCs (PKCα and PKCβ) but not by novel PKCs (PKCδ and PKCε). By replacing the phosphorylated residue with either unphosphorylatable or phosphomimetic residues (S24A and S24E), we found that this phosphorylation event is necessary and sufficient to mediate the PP2A switch, which ultimately induces AKT inactivation, and a robust JNK-dependent apoptosis. Conclusion Our results show that the PP2A switch is induced by PKC-mediated phosphorylation of Ser24-PP2Ac and that this phosphorylation leads to apoptosis upon GqPCR induction of various cells. We propose that this mechanism may provide an unexpected way to treat some cancer types or problems in the endocrine machinery.

Abstract The Thermomyces lanuginosus lipase (TLLs) was successfully immobilized within a novel hydrogel matrix through a two-step crosslinking method. TLLs were initially crosslinked through the Schiff base reaction by oxidized carboxymethyl cellulose (OCMC). The water-soluble OCMC@TLLs complex was subsequently crosslinked by carboxymethyl chitosan (CMCSH) in a microfluidic apparatus to form the CMCHS/OCMC@TLLs microspheres. The CD (Circular Dichroism, CD) and FT-IR (Fourier Transform infrared spectroscopy, FT-IR) spectra demonstrated that the crosslinking of TLLs with OCMC resulted in a less significant impact on their structure compared to that with glutaraldehyde. CMCHS/OCMC@TLLs showed decreased catalytic performance due to the mass transfer resistance, while its thermal stability was greatly improved. The CMCHS/OCMC@TLLs were used to catalyze the lauroylation of arbutin in tetrahydrofuran. After 12 h of reaction under optimal conditions, the yield of 6′-O-lauryl arbutin reached an impressive 92.12%. The prepared 6′-O-lauryl arbutin has high lipophilicity and exhibits similar tyrosinase inhibitory activity and higher antioxidant activity compared to its parent compound. Graphical Abstract

Abstract Ulva is one of the main green algae causing green tide disasters. Ulvan is the primarily component polysaccharide of the cell wall of Ulva and its complex structure and monosaccharide composition resulted in various biological activities. However, the high-value and effective utilization of extracted ulvan have been obstructed by limitations ranging from large molecular weight and low solubility to poor bioavailability. Ulva oligosaccharide obtained by degrading ulvan can not only ideally retain the various biological activities of ulvan very well but also effectively solve the problems of low solubility and poor bioavailability. The preparation and biological activity studies of ulvan and Ulva oligosaccharides have become a hot spot in the field of marine biological resources development research. At present, the comprehensive reviews of ulvan and Ulva oligosaccharides are still scarce. What are overviewed in this paper are the chemical composition, structure, extraction, and purification of ulvan and Ulva oligosaccharides, where research progress on the biological activities of ulvan and Ulva oligosaccharides is summarized and prospected. A theoretical and practical basis has been provided for further research on ulvan and Ulva oligosaccharides, as well as the high-value development and effective utilization of marine algae resources. Graphical Abstract

Rapid extraction and concentration systems based on green materials such as cellulose or lignin are promising. However, there is still a need to optimize the material properties and production processes. Unlike conventional cellulose or lignin sorbent materials, aquatic reed root cells can concentrate external organic pollutants in the water and accumulate them in the plant. Inspired by this, a new nanocellulose–lignin aerogel (NLAG) was designed, in which nanocellulose was used as a substrate and lignin and polyamide epoxy chloropropane were used to crosslink cellulose in order to enhance the strength of the NLGA, resulting in good mechanical stability and water–oil amphiphilic properties. In practical applications, the organic membrane on the NLAG can transport organic pollutants from water to the NLAG, where they are immobilized. This is evidenced by the fact that the aerogel can remove more than 93% of exogenous phenol within a few minutes, highly enriching it inside. In addition, the aerogel facilitates filtration and shape recovery for reuse. This work establishes a novel biopolymer–aerogel-based extraction system with the advantages of sustainability, high efficiency, stability, and easy detachability, which are hard for the traditional adsorbent materials to attain.

Summary: Ferroptosis is a form of regulated cell death with roles in degenerative diseases and cancer. Excessive iron-catalyzed peroxidation of membrane phospholipids, especially those containing the polyunsaturated fatty acid arachidonic acid (AA), is central in driving ferroptosis. Here, we reveal that an understudied Golgi-resident scaffold protein, MMD, promotes susceptibility to ferroptosis in ovarian and renal carcinoma cells in an ACSL4- and MBOAT7-dependent manner. Mechanistically, MMD physically interacts with both ACSL4 and MBOAT7, two enzymes that catalyze sequential steps to incorporate AA in phosphatidylinositol (PI) lipids. Thus, MMD increases the flux of AA into PI, resulting in heightened cellular levels of AA-PI and other AA-containing phospholipid species. This molecular mechanism points to a pro-ferroptotic role for MBOAT7 and AA-PI, with potential therapeutic implications, and reveals that MMD is an important regulator of cellular lipid metabolism.

Objective: This study aimed to select a suitable drying method for preserved yellow peach. Methods: Preserved yellow peach was prepared by hot air drying (HAD), microwave-hot air drying (MHD) and vacuum freeze drying (VFD). Its physical properties, nutritional components, sensory and aroma indexes were studied. Results: Compared with traditional HAD, the efficiency of MHD was increased by 56.1%, and the preserved fruit was significantly better than HAD in color, nutritional composition, sensory evaluation, aroma components and content; The preserved yellow peach fruit by VFD has the best color, the strongest rehydration, the best tissue structure integrity of the product, the highest contents of nutrients vitamin C, and total phenol and titratable acid, and the highest sensory score is 91.58. It has the most aroma components and contents, and uniquely has two aroma components: 2-hexenol and trans-2-hexenal. Conclusion: Compared with HAD, the preserved yellow peach fruit prepared by MHD is better in all aspects, and MHD is more suitable for the production of common preserved yellow peach fruit. The quality of preserved yellow peach fruit prepared by VFD is the best. However, due to the high-energy and -cost consumption, it is recommended to produce high-quality preserved fruit.

Neutralisation assays are key to understanding immune responses to SARS-CoV-2 infection or vaccination. Here, the authors report a surrogate virus neutralization assay called Neu-SATiN, which measures neutralization directly from sera, and allows easy adaptation to variant-specific testing.

Abstract Background G protein-coupled receptors (GPCRs) usually regulate cellular processes via activation of intracellular signaling pathways. However, we have previously shown that in several cell lines, GqPCRs induce immediate inactivation of the AKT pathway, which leads to JNK-dependent apoptosis. This apoptosis-inducing AKT inactivation is essential for physiological functions of several GqPCRs, including those for PGF2α and GnRH. Methods Here we used kinase activity assays of PI3K and followed phosphorylation state of proteins using specific antibodies. In addition, we used coimmunoprecipitation and proximity ligation assays to follow protein–protein interactions. Apoptosis was detected by TUNEL assay and PARP1 cleavage. Results We identified the mechanism that allows the unique stimulated inactivation of AKT and show that the main regulator of this process is the phosphatase PP2A, operating with the non-canonical regulatory subunit IGBP1. In resting cells, an IGBP1-PP2Ac dimer binds to PI3K, dephosphorylates the inhibitory pSer608-p85 of PI3K and thus maintains its high basal activity. Upon GqPCR activation, the PP2Ac-IGBP1 dimer detaches from PI3K and thus allows the inhibitory dephosphorylation. At this stage, the free PP2Ac together with IGBP1 and PP2Aa binds to AKT, causing its dephosphorylation and inactivation. Conclusion Our results show a stimulated shift of PP2Ac from PI3K to AKT termed “PP2A switch” that represses the PI3K/AKT pathway, providing a unique mechanism of GPCR-stimulated dephosphorylation. Video Abstract

Understanding customer needs is of great significance to enhance service quality and competitive advantage. However, for the tourism industry, it is still unclear how to mine service improvement strategies from tourist-generated online reviews. This paper aims to develop a data-driven approach to conduct a fine-grained dimension analysis of customer satisfaction with tourism services. First, this paper uses Latent Dirichlet Allocation to explore the key dimensions of tourist satisfaction from online reviews. Next, based on the Chinese sentiment dictionary, tourists’ emotional attitudes towards each service dimension can be identified. Then, the backpropagation neural network is used to measure the complex relationship between tourists’ sentiment orientations towards different dimensions and their satisfaction. Finally, according to the improved Kano model, multi-dimensional attribute classification is realized to support the strategic analysis of tourism service quality improvement. The proposed method is empirically verified through a real tourism review dataset. The results exhibit the theoretical and practical implications of our method.

Summary: Met is an oncogene aberrantly activated in multiple cancers. Therefore, to better understand Met biology and its role in disease we applied the Mammalian Membrane Two-Hybrid (MaMTH) to generate a targeted interactome map of its interactions with human SH2/PTB-domain-containing proteins. We identified thirty interaction partners, including sixteen that were previously unreported. Non-small cell lung cancer (NSCLC)-focused functional characterization of a Met-interacting protein, BLNK, revealed that BLNK is a positive regulator of Met signaling, and modulates localization, including ligand-dependent trafficking of Met in NSCLC cell lines. Furthermore, the interaction between Met and GRB2 is increased in the presence of BLNK, and the constitutive interaction between BLNK and GRB2 is increased in the presence of active Met. Tumor phenotypical assays uncovered roles for BLNK in anchorage-independent growth and chemotaxis of NSCLC cell lines. Cumulatively, this study provides a Met-interactome and delineates a role for BLNK in regulating Met biology in NSCLC context.

Resibufogenin (RB) is a major active ingredient in the traditional Chinese medicine Chansu and has garnered considerable attention for its efficacy in the treatment of cancer. However, the anticancer effects and underlying mechanisms of RB on glioblastoma (GBM) remain unknown. Here, we found that RB induced G2/M phase arrest and inhibited invasion in a primary GBM cell line, P3#GBM, and two GBM cell lines, U251 and A172. Subsequently, we demonstrated that RB-induced G2/M phase arrest occurred through downregulation of CDC25C and upregulation of p21, which was caused by activation of the MAPK/ERK pathway, and that RB inhibited GBM invasion by elevating intercellular Ca2+ to suppress the Src/FAK/Paxillin focal adhesion pathway. Intriguingly, we confirmed that upon RB binding to ATP1A1, Na+-K+-ATPase was activated as a receptor and then triggered the intracellular MAPK/ERK pathway and Ca2+-mediated Src/FAK/Paxillin focal adhesion pathway, which led to G2/M phase arrest and inhibited the invasion of GBM cells. Taken together, our findings reveal the antitumor mechanism of RB by targeting the ATP1A1 signaling cascade and two key signaling pathways and highlight the potential of RB as a new class of promising anticancer agents.

Serological tests are important diagnostic and disease surveillance tools for addressing the COVID-19 pandemic. Here, the authors present a tri-part Nanoluciferase based assay to detect antibodies against SARS-CoV-2.

The enzymatic degradation of seaweed polysaccharides is gaining interest for its potential in the production of functional oligosaccharides and fermentable sugars. Herein, a novel alginate lyase, AlyRm3, was cloned from a marine strain, Rhodothermus marinus DSM 4252. The AlyRm3 showed optimal activity (37,315.08 U/mg) at 70 °C and pH 8.0, with the sodium alginate used as a substrate. Noticeably, AlyRm3 was stable at 65 °C and also exhibited 30% of maximal activity at 90 °C. These results indicated that AlyRm3 is a thermophilic alginate lyase that efficiently degrades alginate at high industrial temperatures (>60 °C). The FPLC and ESI−MS analyses suggested that AlyRm3 primarily released disaccharides and trisaccharides from the alginate, polyM, and polyG in an endolytic manner. In the saccharification process of sodium alginate (0.5%, w/v), the AlyRm3 yielded numerous reducing sugars (1.73 g/L) after 2 h of reaction. These results indicated that AlyRm3 has a high enzymatic capacity for saccharifying the alginate, and could be used to saccharify the alginate biomass before the main fermentation process for biofuels. These properties make AlyRm3 a valuable candidate for both fundamental research and industrial applications.

Abstract Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a chloride and bicarbonate channel in secretory epithelia with a critical role in maintaining fluid homeostasis. Mutations in CFTR are associated with Cystic Fibrosis (CF), the most common lethal autosomal recessive disorder in Caucasians. While remarkable treatment advances have been made recently in the form of modulator drugs directly rescuing CFTR dysfunction, there is still considerable scope for improvement of therapeutic effectiveness. Here, we report the application of a high‐throughput screening variant of the Mammalian Membrane Two‐Hybrid (MaMTH‐HTS) to map the protein–protein interactions of wild‐type (wt) and mutant CFTR (F508del), in an effort to better understand CF cellular effects and identify new drug targets for patient‐specific treatments. Combined with functional validation in multiple disease models, we have uncovered candidate proteins with potential roles in CFTR function/CF pathophysiology, including Fibrinogen Like 2 (FGL2), which we demonstrate in patient‐derived intestinal organoids has a significant effect on CFTR functional expression.

Efficient lubrication is the key to reducing tool friction and wear, decreasing cutting force and enhancing tool cutting performance. To find a more efficient way to improve the lubrication performance of the tools in high-speed cutting hard-cutting materials, YG6X cemented carbide milling inserts textured with micro-groove parallel to the main cutting edge and Fe3O4 nanofluids with a mass concentration of 0.5% were fabricated, the friction and wear tests and contact angle tests were carried out to investigate the coupling lubrication effects of micro-textures and nanofluids. Results obtained in this work indicated that the friction coefficient of micro-texture sample under nanofluids lubrication was reduced by 92.2% when compared with the un-textured sample under dry friction, and the contact angle of the nanofluids droplet on micro-texture surface was 72.1% lower than that of the emulsion droplet on un-textured surface. Based on the results, a series of experiments were further carried out on high-speed milling Ti6Al4V using micro-texture milling cutter under the nanofluids lubrication. The results showed that cutting forces of micro-texture milling cutters under nanofluids lubrication were reduced by 19.1%, 30.8%, 37.7% and 30.0% when compared with un-textured milling cutters under emulsion lubrication at the spindle speeds of 1000r/min, 2000r/min, 3000r/min and 4000r/min.

Protein-protein interactions are fundamental to the regulation of protein activity and cellular phyisology. Here the authors present Split Intein-Mediated Protein Ligation, which uses bait and prey proteins fused to intein fragments to generate single intact proteins upon interaction.

The hydrodynamic characteristic of transonic motion projectiles with different head diameters are investigated by numerical simulation. Compressibility effect in liquid-phase water are modeled using the Tait state equation. The result shows that with increasing of velocity the compression waves transfer to shock waves，which cause the significant increasing of pressure and decreasing the dimensions of supercavities. While the increasing of head diameter, the thickness, the vapor volume fraction and the drag coefficient of supercavities are all enhanced, which is conducive to the stability of transonic-speed projectiles. The cavity dynamics of the different head projectiles are compared, and the results shows when Mach number is in high region, the truncated cone head projectile is enveloped by a cavity which results in less drag and better stability.

Alginate lyases with unique biochemical properties have irreplaceable value in food and biotechnology industries. Herein, the first new hybrid action mode Thalassotalea algicola-derived alginate lyase gene (TAPL7A) with both psychrophilic and cold-tolerance was cloned and expressed heterologously in E. coli. With the highest sequence identity (43%) to the exolytic alginate lyase AlyA5 obtained from Zobellia galactanivorans, TAPL7A was identified as a new polysaccharide lyases family 7 (PL7) alginate lyase. TAPL7A has broad substrate tolerance with specific activities of 4186.1 U/mg, 2494.8 U/mg, 2314.9 U/mg for polyM, polyG, and sodium alginate, respectively. Biochemical characterization of TAPL7A showed optimal activity at 15 °C, pH 8.0. Interestingly, TAPL7A exhibits both extreme psychrophilic and cold tolerance, which other cold-adapted alginate lyase do not possess. In a wide range of 5–30 °C, the activity can reach 80–100%, and the residual activity of more than 70% can still be maintained after 1 h of incubation. Product analysis showed that TAPL7A adopts a hybrid endo/exo-mode on all three substrates. FPLC and ESI-MS confirmed that the final products of TAPL7A are oligosaccharides with degrees of polymerization (Dps) of 1–2. This study provides excellent alginate lyase candidates for low-temperature environmental applications in food, agriculture, medicine and other industries.

Abstract Background The alginate oligosaccharides have been widely used in agriculture, medicine, and food industries due to their versatile physiological functions such as antioxidant, anticoagulant, and antineoplastic activities. The bifunctional alginate lyases can degrade the alginate polysaccharide more efficiently into alginate oligosaccharides. Therefore, it is crucial to discover new bifunctional alginate lyase for alginate oligosaccharide production. Results Herein, a novel bifunctional alginate lyase FsAlgB was cloned and identified from deep-sea bacterium Flammeovirga sp. NJ-04, which exhibited broad substrate specificity and the highest activity (1760.8 U/mg) at pH 8.0 and 40 °C. Furthermore, the K m values of FsAlgB towards polyG (0.69 mM) and polyMG (0.92 mM) were lower than that towards sodium alginate (1.28 mM) and polyM (2.06 mM). Recombinant FsAlgB was further characterized as an endolytic alginate lyase, and it can recognize the tetrasaccharide as the minimal substrate and cleave the glycosidic bonds between the subsites of − 3 and + 1. Conclusion This study provided extended insights into the substrate recognition and degrading pattern of alginate lyases with broad substrate specificity.

Background/Aims: Gq protein-coupled receptors (GqPCRs) regulate various cellular processes including mainly proliferation and differentiation. In a previous study, we found that in prostate cancer cells, the GqPCR of GnRH induces apoptosis by reducing the PKC-dependent AKT activity and elevating JNK phosphorylation. Since it was thought that GqPCR induces mainly activation of AKT, we undertook to examine how general is this phenomenon and understand its signaling. Methods: We used various cells to follow the phosphorylation of signaling components using western blotting. Results: In a screen of 21 cell lines, we found that PKC activation results in the reduction of AKT activity, which correlates nicely to JNK activation and in some cases to apoptosis. To further understand the signaling pathways involved in this stimulation, we studied in detail the SVOG-4O and αT3-1 cells. We found that PGF2α and GnRH agonist (GnRH-a) indeed induce significant Gq- and PKC- dependent apoptosis in these cells. This is mediated by two signaling branches downstream of PKC, which converge at the level of MLK3 upstream of JNK. One branch consists on c-Src activation of the JNK cascade and the second involves reduction of AKT activity that alleviates its inhibitory effect on MLK3, to allow the flow of the c-Src signal to JNK. At the MAPKK level, we found that the signal is transmitted by MKK7 and not MKK4. Conclusion: Our results present a general mechanism that mediates a GqPCR-induced, death receptors-independent, apoptosis in physiological, as well as cancer-related systems.

The high-valued utilization of Ulva (previously known as Enteromorpha) bioresources has drawn increasing attention due to the periodic blooms of world-wide green tide. The polysaccharide is the main functional component of Ulva and exhibits various physiological activities. The Ulva oligosaccharide as the degradation product of polysaccharide not only possesses some obvious activities, but also possesses excellent solubility and bioavailability. Both Ulva polysaccharides and oligosaccharides hold promising potential in the food industry as new functional foods or food additives. Studies on Ulva polysaccharides and oligosaccharides are increasing and have been the focus of the marine bioresources field. However, the comprehensive review of this topic is still rare and do not cover the recent advances of the structure, isolation, preparation, activity and applications of Ulva polysaccharides and oligosaccharides. This review systematically summarizes and discusses the recent advances of chemical composition, extraction, purification, structure, and activity of Ulva polysaccharides as well as oligosaccharides. In addition, the potential applications as new functional food and food additives have also been considered, and these will definitely expand the applications of Ulva oligosaccharides in the food and medical fields.

Gap junctional intercellular communication (GJIC) composed of connexin proteins is considered vital to cancer onset and progression since 50 years ago based on Lowenstein and Kano's works, however altered expression of connexins is still a lesser known “hallmark” of cancer. Although many studies support the hypothesis that connexins are tumor suppressors, recent evidence indicates that, in some tumor types including glioma, they may play contradictory role in some specific stages of tumor progression. We thus conduct a meta-analysis to evaluate the prognostic role of Cx43 in glioma for the unanswered questions that whether Cx43 is a beneficial or insalubrity factor for glioma. Eight studies with 1,706 patients were included for meta-analysis. The results showed that Cx43 expression was a clearly negative factor with tumor grades (I2 = 34%, P < 0.001) and beneficial for OS (n = 3, HR 2.62, 95%CI 1.47–4.68; P = 0.001). Subgroup analysis also found that Cx43 had different expression in Asian young patients vs. other groups. In conclusion, this article summarize the prognostic value of Cx43 and offer a clinical evidence for the notion that Cx43 is generally a tumor suppressor and beneficial for the patients' survival time.

Everyone likes Porsche but few can afford it. Budget constraints always play a critical role in a customer’s decision-making. The literature disproportionally focuses on how firms can induce customer valuations toward the product, but does not address how to assess the influence of budget constraints. We study these questions in the context of a referral reward program (RRP). RRP is a prominent marketing strategy that utilizes recommendations passed from existing customers to their friends and effectively stimulates word of mouth (WoM). We build a stylized game-theoretical model with a nested Stackelberg game involving three players: a firm, an existing customer, and a potential customer who is a friend of the existing customer. The budget is the friend’s private information. We show that RRPs might be optimal when the friend has either a low or a high valuation, but they work differently in each situation because of the budget. Furthermore, there are two budget thresholds, a fixed one and a variable one, which limit a firm’s ability to use rewards.

Alginate oligosaccharides with different degrees of polymerization (DPs) possess diverse physiological activities. Therefore, in recent years, increasing attention has been drawn to the use of enzymes for the preparation of alginate oligosaccharides for food and industrial applications. Previously, we identified and characterized a novel bifunctional alginate lyase Aly7A, which can specifically release trisaccharide from three different substrate types with a unique degradation pattern. Herein, we investigated its degradation pattern by modular truncation and molecular docking. The results suggested that Aly7A adopted a unique action mode towards different substrates with the substrate chain sliding into the binding pocket of the catalytic domain to position the next trisaccharide for cleavage. Deletion of the Aly7A carbohydrate binding module (CBM) domain resulted in a complex distribution of degradation products and no preference for trisaccharide formation, indicating that the CBM may act as a “controller” during the trisaccharide release process. This study further testifies CBM as a regulator of product distribution and provides new insights into well-defined generation of alginate oligosaccharides with associated CBMs.

SERINC5 is a host restriction factor that impairs infectivity of HIV-1 and other primate lentiviruses and is counteracted by the viral accessory protein Nef. However, the importance of SERINC5 antagonism for viral replication and cytopathicity remained unclear. Here, we show that the Nef protein of the highly divergent SIVcol lineage infecting mantled guerezas (Colobus guereza) is a potent antagonist of SERINC5, although it lacks the CD4, CD3 and CD28 down-modulation activities exerted by other primate lentiviral Nefs. In addition, SIVcol Nefs decrease CXCR4 cell surface expression, suppress TCR-induced actin remodeling, and counteract Colobus but not human tetherin. Unlike HIV-1 Nef proteins, SIVcol Nef induces efficient proteasomal degradation of SERINC5 and counteracts orthologs from highly divergent vertebrate species, such as Xenopus frogs and zebrafish. A single Y86F mutation disrupts SERINC5 and tetherin antagonism but not CXCR4 down-modulation by SIVcol Nef, while mutation of a C-proximal di-leucine motif has the opposite effect. Unexpectedly, the Y86F change in SIVcol Nef had little if any effect on viral replication and CD4+ T cell depletion in preactivated human CD4+ T cells and in ex vivo infected lymphoid tissue. However, SIVcol Nef increased virion infectivity up to 10-fold and moderately increased viral replication in resting peripheral blood mononuclear cells (PBMCs) that were first infected with HIV-1 and activated three or six days later. In conclusion, SIVcol Nef lacks several activities that are conserved in other primate lentiviruses and utilizes a distinct proteasome-dependent mechanism to counteract SERINC5. Our finding that evolutionarily distinct SIVcol Nefs show potent anti-SERINC5 activity supports a relevant role of SERINC5 antagonism for viral fitness in vivo. Our results further suggest this Nef function is particularly important for virion infectivity under conditions of limited CD4+ T cell activation.

To obtain a global picture of how protein phosphatases are involved in receptor tyrosine kinase (RTK) signaling, we mapped the RTK-phosphatase interactome. Analyses of selected interactions revealed detailed mechanisms of their actions. This study provides new knowledge to better understand cancer development and to identify novel therapeutic targets.

Abstract G‐protein‐coupled receptors (GPCRs) are the largest family of integral membrane receptors with key roles in regulating signaling pathways targeted by therapeutics, but are difficult to study using existing proteomics technologies due to their complex biochemical features. To obtain a global view of GPCR‐mediated signaling and to identify novel components of their pathways, we used a modified membrane yeast two‐hybrid (MYTH) approach and identified interacting partners for 48 selected full‐length human ligand‐unoccupied GPCRs in their native membrane environment. The resulting GPCR interactome connects 686 proteins by 987 unique interactions, including 299 membrane proteins involved in a diverse range of cellular functions. To demonstrate the biological relevance of the GPCR interactome, we validated novel interactions of the GPR37, serotonin 5‐HT4d, and adenosine ADORA2A receptors. Our data represent the first large‐scale interactome mapping for human GPCRs and provide a valuable resource for the analysis of signaling pathways involving this druggable family of integral membrane proteins.

A considerable amount of marketing research has reported that consumers are more saliently influenced by friends (strong social ties) than by acquaintances and strangers (weak social ties). To shed light on the neural and psychological processes underlying such phenomenon, in this study we designed an amended S1-S2 paradigm (product-[reviewer-review]) that is based on realistic consumer purchase experiences. After incoming all given information (product, reviewer, review), participants were required to state their purchase intentions. The neurocognitive and emotional processes related to friend and stranger stimuli were delineated to suggest how social ties influence consumers during their shopping processes. Larger P2 (fronto-central scalp areas) and P3 (central and posterior-parietal scalp areas) components under stranger condition were elicited successfully. These findings demonstrate that the cognitive and emotional processing of friend and stranger stimuli occurs at stages of neural activity, and can be indicated by the P2 and P3 components. Electrophysiological data also support the hypothesis that different neural and emotional processing magnitude and strength underlie friend and stranger effect in the context of consumer purchase. During this process, the perception of stimuli evoked P2, subsequently emotional processing and attention modulation were activated and indicated by P2 and P3. The friend dominated phenomenon can be interpreted as the result of distinctive neurocognitive and emotional processing magnitude, which suggests that psychological and emotional factors can guide consumer decision making. This study consolidates that event related potential (ERP) methodology is likely to be a more sensitive method for investigating consumer behaviors. From the perspectives of management and marketing, our findings show that the P2 and P3 components can be employed as an indicator to probe the influential factors of consumer purchase intentions.

Alginate lyases have been widely used to prepare alginate oligosaccharides in food, agricultural, and medical industries. Therefore, discovering and characterizing novel alginate lyases with excellent properties has drawn increasing attention. Herein, a novel alginate lyase FsAlyPL6 of Polysaccharide Lyase (PL) 6 family is identified and biochemically characterized from Flammeovirga sp. NJ-04. It shows highest activity at 45 °C and could retain 50% of activity after being incubated at 45 °C for 1 h. The Thin-Layer Chromatography (TLC) and Electrospray Ionization Mass Spectrometry (ESI-MS) analysis indicates that FsAlyPL6 endolytically degrades alginate polysaccharide into oligosaccharides ranging from monosaccharides to pentasaccharides. In addition, the action pattern of the enzyme is also elucidated and the result suggests that FsAlyPL6 could recognize tetrasaccharide as the minimal substrate and cleave the glycosidic bonds between the subsites of −1 and +3. The research provides extended insights into the substrate recognition and degradation pattern of PL6 alginate lyases, which may further expand the application of alginate lyases.

Currently, wetland stability is under threat due to the joint effects of global climate change and human activity, especially in lakes. Hence, it is necessary to evaluate the health status of wetland ecosystems such as lakes, identify the variables causing the wetland degradation and work to protect the wetlands from the identified variables in the future. Based on fourteen high-resolution autumn remote sensing images from 1989–2013, the classification characteristics and spatial distribution patterns of wetland landscapes in Poyang Lake were studied through quantitative interpretation technology. An established health assessment index system named the EHCI (Ecological Health Comprehensive Index) was used to assess the health status of Poyang Lake. Additionally, the relationship between water regime and health status of wetland landscape distribution of Poyang Lake were investigated by multivariate statistical analysis. The results demonstrated: (1) The total area of three first level (or six second level) types of wetland landscapes showed a stable status, which was long-term maintaining at about 3026 km2 from 1989–2013. (2) The water area shows a downward trend, while the areas of vegetation and land-water transition zone show upward trends. (3) The proposed EHCI of the Poyang Lake wetland presented a downward trend. According to the EHCI results from 1989–2013, the health status of Poyang Lake wetland was healthy for two years, unhealthy for four years and sub-healthy for eight years. (4) The water level fluctuation greatly affected the EHCI, and the effect became greater as the water level increased. These results contribute to the understanding of specific effects of hydrological process on the health status of the Poyang Lake wetland. In addition they provide a scientific reference for the maintenance of stable ecosystem functions of the seasonal freshwater lake. These results contribute to the understanding of specific effects of hydrological process on the health status of the Poyang Lake. In addition they provide a scientific reference for the maintenance of stable ecosystem functions of the seasonal freshwater lake.

This study aimed to explore the effects of a lexical approach to Chinese English major students' reading comprehension ability as well as students' opinions of this approach. A quasiexperimental study was carried out in a Normal University in southern China for 11 weeks. An intact class of 45 first-year English major students at intermediate level participated in the study. They were instructed using a lexical approach to study four long reading texts. The instruments included reading comprehension tests, a questionnaire, and a semistructured interview. The results showed that students made much progress in reading comprehension after the treatment and they had positive opinions of the lexical approach. It can be concluded from this that the lexical approach had positive impacts on Chinese English major students' reading comprehension ability and it suggests that this approach could be implemented in English language teaching and learning for English majors in higher education.

Enzymatic preparation of alginate oligosaccharides with versatile bioactivities by alginate lyases has attracted increasing attention due to its featured characteristics, such as wild condition and specific products. In this study, AlgNJ-07, a novel polyM-specific alginate lyase with high specific activity and pH stability, has been purified from the newly isolated marine bacterium Serratia marcescens NJ-07. It has a molecular weight of approximately 25 kDa and exhibits the maximal activity of 2742.5 U/mg towards sodium alginate under 40 °C at pH 9.0. Additionally, AlgNJ-07 could retain more than 95% of its activity at pH range of 8.0–10.0, indicating it possesses excellent pH-stability. Moreover, it shows high activity and affinity towards polyM block and no activity to polyG block, which suggests that it is a strict polyM-specific alginate lyase. The degradation pattern of AlgNJ-07 has also been explored. The activity of AlgNJ-07 could be activated by NaCl with a low concentration (100–300 mM). It can be observed that AlgNJ-07 can recognize the trisaccharide as the minimal substrate and hydrolyze the trisaccharide into monosaccharide and disaccharide. The TLC and ESI-MS analysis indicate that it can hydrolyze substrates in a unique endolytic manner, producing not only oligosaccharides with Dp of 2–5 but also a large fraction of monosaccharide. Therefore, it may be a potent tool to produce alginate oligosaccharides with lower Dps (degree of polymerization).

Considering the advantages of distribution and maximum parallelism of membrane computing and availability of discrete Morse theory to deal with discrete structure, in this paper, combining discrete Morse theory and membrane computing, a novel membrane structure—P system with chain structure, is proposed, which is constructed on the basis of discrete gradient vector path of the discrete Morse theory. At the theoretical level, due to its unique chain structure, compared with traditional P system, its structure, object, and rule are described in details. In the practical aspect, a specific application example, P chain system for arithmetic operation, is presented to demonstrate the superiority, computational efficiency, and ability of P system with chain structure. Moreover, a simulation system of arithmetic operations based on P chain system is designed, giving a visual display of the implementation of P chain system for arithmetic operation, and verifying the feasibility and effectiveness of P chain system.

Emergency incidents, including natural disasters, terrorist attacks, public health outbreaks, and industrial and mining accidents, and so forth, result in severe human casualties and property losses. Emergency facilities, which provide relief materials and services, play an important role in rescue management. The decision of where to locate the emergency rescue facilities is very important, as it determines the efficiency and effectiveness of the emergency management process. This paper develops a modified -median problem model that accounts for rescue time limitations. A variable neighbourhood search- (VNS-) based algorithm is developed for the model considered. The modified VNS algorithm exhibits good performance on -median benchmark problems. A case from Western China is studied, and a reasonable location decision is then made for emergency rescue facilities using the modified VNS algorithm. The paper also compares the results with and without considering the rescue time limitation.

The ERK5 cascade is a MAPK pathway that transmits both mitogenic and stress signals, yet its mechanism of activation is not fully understood. Using intracellular calcium modifiers, we found that ERK5 activation by EGF is inhibited both by the depletion and elevation of intracellular calcium levels. This calcium effect was found to occur upstream of MEKK2, which is the MAP3K of the ERK5 cascade. Co-immunoprecipitation revealed that EGF increases MEKK2 binding to the adaptor protein Lad1, and this interaction was reduced by the intracellular calcium modifiers, indicating that a proper calcium concentration is required for the interactions and transmission of EGF signals to ERK5. In vitro binding assays revealed that the proper calcium concentration is required for a direct binding of MEKK2 to Lad1. The binding of these proteins is not affected by c-Src-mediated phosphorylation on Lad1, but slightly affects the Tyr phosphorylation of MEKK2, suggesting that the interaction with Lad1 is necessary for full Tyr phosphorylation of MEKK2. In addition, we found that changes in calcium levels affect the EGF-induced nuclear translocation of MEKK2 and thereby its effect on the nuclear ERK5 activity. Taken together, these findings suggest that calcium is required for EGF-induced ERK5 activation, and this effect is probably mediated by securing proper interaction of MEKK2 with the upstream adaptor protein Lad1.