Search Results (151)

Spurred by the enormous therapeutic success of glucagon-like peptide-1 receptor (GLP-1R) agonists (GLP1-RAs) against diabetes and obesity, glucagon family receptor pharmacology has garnered a tremendous amount of interest. Glucagon family receptors, e.g., the glucagon receptor itself (GCGR), the GLP-1R, and the glucose-dependent insulinotropic peptide receptor (GIPR), belong to the incretin receptor superfamily, i.e., receptors that increase blood glucose-dependent insulin secretion. All incretin receptors are class B1 G protein-coupled receptors (GPCRs), coupling to the G_s type of heterotrimeric G proteins which activates adenylyl cyclase (AC) to produce cyclic adenosine monophosphate (cAMP). Most GPCRs undergo desensitization, i.e., uncouple from G proteins and internalize, thanks to interactions with the βarrestins (arrestin-2 and -3). Since the βarrestins can also mediate their own G protein-independent signaling, any given GPCR can theoretically signal (predominantly) either via G proteins or βarrestins, i.e., be a G protein- or βarrestin-“biased” receptor, depending on the bound ligand. A plethora of experimental evidence suggests that the GLP-1R does not undergo desensitization in physiologically relevant tissues in vivo, but rather, it produces robust and prolonged cAMP signals. A particular property of constant cycling between the cell membrane and caveolae/lipid rafts of the GLP-1R may underlie its lack of desensitization. In contrast, GIPR signaling is extensively mediated by βarrestins and the GIPR undergoes significant desensitization, internalization, and downregulation, which may explain why both agonists and antagonists of the GIPR exert the same physiological effects. Here, we discuss this evidence and make a case for the GLP-1R being a phenotypically or functionally G_s-selective receptor. We also discuss the implications of this for the development of GLP-1R poly-ligands, which are increasingly pursued for the treatment of obesity and other diseases. Full article

Hyperekplexia (OMIM 149400), a sensorimotor syndrome of perinatal clinical relevance, causes newborns to display an energic startle reflex in response to certain trivial stimuli. This condition can be lethal due to apnea episodes. The disease is caused by a blockade of glycinergic neurotransmission. Glycinergic interneurons preserve their identity by the activity of the surface glycine transporter GlyT2, which supplies glycine to presynaptic terminals to maintain glycine content in synaptic vesicles. Loss-of-function mutations in the GlyT2 gene (SLC6A5) cause a presynaptic form of human hyperekplexia. Here, we describe a new GlyT2 variant found in an infantile patient diagnosed with hyperekplexia. A missense mutation in the open reading frame of the GlyT2 gene inherited in homozygosity caused the substitution G449E in a residue highly conserved across the phylogenetic scale. The sequences of the glycine receptor genes GLRA1 and GLRB did not show abnormalities. We expressed the recombinant GlyT2 variant in heterologous cells and analyzed its pathogenic mechanism. The transporter was totally inactive, behaving as a bona fide loss-of-function mutant. Furthermore, the mutation promoted the abnormal insertion of the protein into the membrane, leading to its large incorporation into lipid rafts. However, there was no apparent alteration of wild-type trafficking upon mutant coexpression, as the mutant was prematurely degraded from the endoplasmic reticulum. Rescue with chemical chaperones was not possible for this mutant. Proteomics demonstrated that the expression of the mutant induced the unfolded protein response and interfered with raft-dependent processes. Therefore, the new variant causes a loss of function regarding GlyT2 activity but a gain of function as a cell proteostasis disturber. Full article

The evaluation of tourism climate comfort is a systematic assessment of the climate conditions of tourist destinations. It is of great significance for improving the tourism experience, promoting the sustainable development of the tourism industry, and protecting the natural environment. In this study, the CiteSpace software was used to conduct a bibliometrics analysis of the study on tourism climate comfort in China, and the conceptual framework of this study was established based on the bibliometrics results. In the conceptual framework, this study firstly summarized eight indicators widely used in the current evaluation of tourism climate comfort. Secondly, four key technical means in the evaluation process were summarized, including geographic information system, analytic hierarchy process, fuzzy comprehensive evaluation, and cluster analysis. And three calculation methods of tourism climate comfort period were summarized, namely number of days with comfortable climate, five-day moving average method, and probability of climate-suitable days. Subsequently, the main application areas of tourism climate comfort evaluation were introduced: (1) exploration of the relationship between climate comfort and tourism activities (i.e., heat/cold-escape tourism, ice-snow tourism, outdoor rafting, coastal tourism, and other types of tourism activities); (2) exploration of the relationship between climate comfort and tourist flow; (3) the response of climate comfort to climate change; and (4) tourism climate regionalization. Finally, the main problems of current research and future development directions were proposed. Full article

Understanding the complex phenomena of interactions between the elements of a combined piled raft foundation (CPRF) is essential for the proper design of such foundations. To evaluate the effects of mutual influence among the CPRF’s elements, a series of long-term measurements of selected physical quantities related to the performance of the foundation were conducted on a building with a frame structure, stiffening walls, and monolithic technology, consisting of seven aboveground stories and one underground story. The analysis distinguishes the real deformations resulting from temperature changes and from stress strains resulting from load changes. The two types of deformations were subjected to further interpretation of only changes in the stress and strain over time. Changes in stress values in the subsoil, as well as strain measurements in the vertical direction of concrete columns, were recorded to assess the load distribution between the CPRF’s components. The numerical analysis results obtained for a fragment of the monitored foundation were compared with actual measurement results to verify the numerical model of interaction between the structure and the soil. Field monitoring and FEA methods were used to compare the long-term deformation analysis, and they helped to minimize the monitoring time. This comparison also served to supplement and simultaneously expand the dataset of test results on a real-world scale. Full article

The rapid regulatory mechanism of light-induced state transitions (STs) in oxygenic photosynthesis is particularly appealing for membrane-based applications. This interest stems from the unique ability of the thylakoid membrane protein cytochrome b₆f (cytb₆f) to increase or decrease its hydrophobic thickness (d_P) in parallel with the reduction or oxidation of the PQ pool induced by changes in light quality. This property appears to be the long-sought biophysical driver behind the reorganizations of membrane proteins during STs. This study decisively advances the hydrophobic mismatch (HMM) model for cytb₆f-driven STs by thoroughly analyzing thirteen X-ray crystal and eight cryo-electron microscopy cytb₆f structures. It uncovers the lipid nanoenvironments that cytb₆f, with different hydrophobic thicknesses, selectively attracts. Under optimal, stationary conditions for photosynthesis in low light, when there is hydrophobic matching between the hydrophobic thicknesses of cytb₆f d_P and that of the bulk thylakoid lipid phase d_L, d_P = d_L, cytb₆f predominantly binds to anionic lipids—several phosphatidylglycerol (PG) molecules and one sulfoquinovosyldiacylglycerol (SQDG) molecule. Upon the induction of the transition to State 2, when d_P increases and induces a positive HMM (d_P > d_L), the neutral, non-bilayer-forming lipid monogalactosyldiacylglycerol (MGDG) replaces some of the bound PGs. Upon the induction of the transition to State 1, when d_P decreases and induces a negative HMM (d_P < d_L), PGs and SQDG detach from their binding sites, and two neutral, bilayer-forming lipids such as digalactosyldiacylglycerol (DGDG) occupy two sites. Additionally, this research uncovers two lipid-mediated signaling pathways from Chla to the center of flexibility, the Phe/Tyr124^fg-loop-suIV residue—one of which involves β-carotene. This study identifies two novel types of lipid raft-like nanodomains that are devoid of typical components, such as sphingomyelin and cholesterol. These findings firmly validate the HMM model and underscore the STs as the first recognized functional process that fully utilizes the unique and evolutionarily conserved composition of just four thylakoid lipid classes. This research contributes to our understanding of membrane dynamics in general and STs in particular. It introduces a novel and simple approach for reversible protein reorganization driven purely by biophysical mechanisms, with promising implications for various membrane-based applications. Full article

The complexes of negatively charged polysaccharides with lipid vesicles have been shown to have applications in medicine, bioremediation, water purification, and construction of nano-biosensors. This article presents research on the formation of these complexes based on the interactions between three types of liposomes, DOPC liposomes (which contain a lipid bilayer in the liquid-disordered (Ld) state), RAFT liposomes (which contain liquid-ordered (Lo) lipid raft domains surrounded by lipids in the Ld state) and SPH–CHL liposomes (which contain a lipid bilayer in the Lo state), and two selected anionic polysaccharides, polysialic acid (PSA) and polygalacturonic acid (PGA). The analysis was conducted using a toluidine blue (TB) probe and the absorption spectroscopy technique. In contrast to DOPC and SPH–CHL liposomes, binding of negatively charged PSA or PGA chains to RAFT liposomes induced a TB absorption maximum shift from 630 nm to 560 nm. The obtained results indicate that toluidine blue can be applied for monitoring the formation of these nano-complexes, and that the boundaries between Ld/Lo domains within membranes in RAFT liposomes can significantly enhance the binding affinity of negatively charged polysaccharides to the lipid bilayer surface. The observed metachromatic shift in TB absorption suggests that negatively charged PSA and PGA chains interact with the Ld/Lo boundaries within RAFT liposome membranes. Full article

Directionally solidified (DS) superalloys have become a primary material choice for turbine blade applications. Due to the complex shape of the blades, certain regions inevitably experience stress axes oriented orthogonally to the crystal growth direction. Therefore, this study explores the creep characteristics of a DS superalloy in different orientations (transverse (T) versus longitudinal (L) with respect to grain growth direction) under intermediate and high temperatures (980 °C and 1070 °C), while simultaneously analyzing their respective deformation mechanisms and microstructural transformation behaviors. Experimental findings reveal pronounced orientation-dependent variations in creep performance, deformation modes, and microstructural development. Notably, the T specimen exhibits higher creep resistance at 980 °C, which can provide a basis for the design of some components that require high creep resistance and maintain small deformation. At 980 °C, L specimens primarily undergo γ′ phase shearing via antiphase boundaries (APBs) pairs, whereas T specimen exhibits APB pairs and superlattice intrinsic stacking faults (SISFs) shearing mechanisms. At 1070 °C, the L specimen exhibits dislocation shearing of γ′ alongside dislocation bypassing of tertiary γ′, while the T specimen demonstrates dislocation climbing within the γ channels. Additionally, the L specimen exhibits significant N-type rafting, while the T specimen shows significant Ostwald ripening characteristics, with an Ostwald ripening rate constant of 1.04 × 10⁻²⁰ m³/h. Full article

Novel amphiphilic block copolymers of N-vinyl pyrrolidone (NVP) and either n-hexyl methacrylate (HMA, PNVP-b-PHMA) or stearyl methacrylate (SMA, PNVP-b-PSMA) were prepared by RAFT polymerization techniques and the sequential addition of monomers starting from the polymerization of NVP and using two different Chain Transfer Agents, CTAs. PNVP-b-PHMA are amorphous block copolymers containing constituent blocks with both high and low Tg values, whereas PNVP-b-PSMA are amorphous–semi-crystalline copolymers. Samples with different molecular weights and compositions were obtained. The copolymers were microphase-separated, but partial mixing was also observed. The presence of the amorphous PNVP block reduced the crystallinity of the PSMA blocks in the PNVP-b-PSMA copolymers. The thermal stability of the blocks was influenced by both constituents. The self-assembly behavior in THF, which is a selective solvent for polymethacrylate blocks, and in aqueous solutions, where PNVP was soluble, was examined. Unimolecular or low-aggregation-number micelles were obtained in THF for both types of samples. On the contrary, high-aggregation-number, spherical, and compact micelles were revealed in aqueous solutions. The increase in the steric hindrance of the side ester group of the polymethacrylate chain led to slightly lower degrees of association. The hydrophobic compound curcumin was efficiently encapsulated within the micellar core of the supramolecular structures in aqueous solutions. Micelles with higher aggregation numbers were more efficient in the encapsulation of curcumin. The results of this study were compared with those obtained from other block copolymers based on PNVP. Full article

By testing the creep properties of a Re/Ru-containing single-crystal alloy specimen and examining the microstructural evolution of the allow at different stages of creep using scanning electron microscopy (SEM) and transmission electron microscopy (TEM), the deformation and damage mechanisms of the alloy under ultra-high temperature conditions were investigated. It was observed that a dislocation network forms before the rafting of the γ′ phase. As creep progresses, this network becomes increasingly dense and complete. Moreover, the dislocation network undergoes a transformation from the <110>-type to the <100>-type configuration, with a hybrid <110>-<100>-type network representing an intermediate state during the transition. Stacking faults were also identified within the γ′ phase, suggesting that the stacking fault energy of this alloy is lower compared to that of other alloys. During creep, dislocations that penetrate the γ′ phase can undergo cross slip from the {111} plane to the {100} plane under applied stress, resulting in the formation of Kear–Wilsdorf (K–W) immobile dislocation locks. These locks hinder further dislocation movement within the γ′ phase. It is concluded that the damage mechanism of the alloy at the later stage of creep under 120 MPa/1160 °C involves initial crack formation at the interface of the twisted raft-like γ/γ′ two-phase structure. As creep continues, the crack propagates in a direction perpendicular to the applied stress axis. Full article

The creep properties of directionally solidified superalloys are largely influenced by the degradation rate of the γ/γ’ microstructure and the dislocation motion, which exhibit distinct mechanisms under varying temperature and stress conditions. In this study, the creep deformation mechanisms and microstructural evolution of a directionally solidified nickel-based superalloy in the longitudinal (L) and transverse (T) orientations at 850 °C are comprehensively investigated. Creep testing and characterization of the dislocation structure revealed superior creep properties in the L direction compared to the T direction. The creep mechanism in the L direction involves the activation of multiple {111}<110> slip systems, shearing the γ’ precipitates through antiphase boundaries (APBs). Conversely, the creep mechanism in the T direction involves the activation of {111}<112> slip systems, shearing the γ’ precipitates through a superlattice intrinsic stacking fault (SISF) and forming slip bands inclined to the stress axis. Aluminum was identified as the controlling element for the γ’ rafting. The longitudinal specimens exhibited P-type rafting due to the activation of multiple slip systems and sufficient plastic strain flow from the dislocation motion. In contrast, the transverse specimens show little rafting due to limited slip system activation. These findings can serve as a reference for better understanding the anisotropy of directionally solidified superalloys and provide a basis for their broader application. Full article

The EFR3 (Eighty-Five Requiring 3) protein and its homologs are rather poorly understood eukaryotic plasma membrane peripheral proteins. They belong to the armadillo-like family of superhelical proteins. In higher vertebrates two paralog genes, A and B were found, each expressing at least 2–3 protein isoforms. EFR3s are involved in several physiological functions, mostly including phosphatidyl inositide phosphates, e.g., phototransduction (insects), GPCRs, and insulin receptors regulated processes (mammals). Mutations in the EFR3A were linked to several types of human disorders, i.e., neurological, cardiovascular, and several tumors. Structural data on the atomic level indicate the extended superhelical rod-like structure of the first two-thirds of the molecule with a typical armadillo repeat motif (ARM) in the N-terminal part and a triple helical motif in its C-terminal part. EFR3s’ best-known molecular function is anchoring the giant phosphatidylinositol 4-kinase A complex to the plasma membrane crucial for cell signaling, also linked directly to the KRAS mutant oncogenic function. Another function connected to the newly uncovered interaction of EFR3A with flotillin-2 may be the participation of the former in the organization and regulation of the membrane raft domain. This review presents EFR3A as an intriguing subject of future studies. Full article

Background: Sorting nexin 19 (SNX19) is important in the localization and trafficking of the dopamine D₁ receptor (D₁R) to lipid raft microdomains. However, the interaction between SNX19 and the lipid raft components caveolin-1 or flotillin-1 and, in particular, their roles in the cellular endocytosis and cell membrane trafficking of the D₁R have not been determined. Methods: Caveolin-1 and flotillin-1 motifs were analyzed by in silico analysis; colocalization was observed by confocal immunofluorescence microscopy; protein-protein interaction was determined by co-immunoprecipitation. Results: In silico analysis revealed the presence of putative caveolin-1 and flotillin-1 binding motifs within SNX19. In mouse and human renal proximal tubule cells (RPTCs), SNX19 was localized mainly in lipid rafts. In mouse RPTCs transfected with wild-type (WT) Snx19, fenoldopam (FEN), a D₁-like receptor agonist, increased the colocalization of SNX19 with caveolin-1 and flotillin-1. FEN also increased the co-immunoprecipitation of SNX19 with caveolin-1 and flotillin-1, effects that were prevented by SCH39166, a D₁-like receptor antagonist. The FEN-mediated increase in the residence of SNX19 in lipid rafts and the colocalization of the D₁R with caveolin-1 and flotilin-1 were attenuated by the deletion of a caveolin-1 (YHTVNRRYREF) (ΔCav1) or a flotillin-1 (EEGPGTETETGLPVS) (ΔFlot1) binding motif. The FEN-mediated increase in intracellular cAMP production was also impaired by the deletion of either the flotillin-1 or caveolin-1 binding motif. Nocodazole, a microtubule depolymerization inhibitor, interfered with the FEN-mediated increase in the colocalization between SNX19 and D₁R. Conclusion: SNX19 contains caveolin-1 and flotillin-1 binding motifs, which play an important role in D₁R endocytosis and signaling. Full article

Research on the extraction of satellite information for the areas of coastal fish cages and rafts is important to quickly grasp the pattern and structure of the coastal fishery aquaculture industry. This study proposes a multi-feature and rule-based object-oriented image classification (MROIC) model, integrating spatial-spectral enhancement techniques with object-based image analysis classification methods. The MROIC model enhances spectral information by constructing ratio bands alongside principal component analysis, subsequently employing rule sets, edge detection algorithms, and comprehensive algorithmic merging techniques. It is applicable to satellite image classification tasks in complex environments, including influence of clouds and vessels. The information of fish cage and raft facilities is extracted via the MROIC model on the southwest coast of Xiapu County, Fujian Province, as an example. The results showed that the MROIC model attained an average total classification accuracy of 90.43% and a Kappa coefficient of 0.80. Extracting the area of fisheries facilities under the influence of clouds and vessels can provide better extraction accuracy and lower omission error. The MROIC model proposed in this study demonstrates high extraction accuracy and strong applicability, offering technical support for government planning in fishery facility areas and aiding in the risk assessment and management efficiency of fishery facility insurance. Full article

Mycoplasma bovis (M. bovis) is capable of causing pneumonia, arthritis, mastitis, and various other ailments in cattle of all age groups, posing a significant threat to the healthy progression of the worldwide cattle industry. The invasion of non-phagocytic host cells serves as a pivotal mechanism enabling M. bovis to evade the immune system and penetrate mucosal barriers, thereby promoting its spread. To investigate the differences in M. bovis invasion into four types of non-phagocytic cells (Madin–Darby bovine kidney (MDBK) cells, embryonic bovine lung (EBL) cells, bovine embryo tracheal (EBTr) cells and bovine turbinate (BT) cells) and further elucidate its invasion mechanism, this study first optimized the experimental methods for M. bovis invasion into cells. Utilizing laser scanning confocal microscopy, transmission electron microscopy, and high-content live-cell imaging systems, the invasion process of M. bovis into four types of non-phagocytic cells was observed. The invasion rates of three different strains of M. bovis (PG45, 07801, 08M) were quantified through the plate counting method. In order to clarify the specific pathway of M. bovis invasion into cells, chlorpromazine (CPZ), amiloride (AMI), and methyl-β-cyclodextrin (M-β-CD) were used to inhibit CLR-mediated clathrin-dependent endocytosis (CDE) pathway, macropinocytosis, and lipid raft pathway, respectively. Subsequently, the invasion rates of PG45 into these four types of cells were measured. Using siRNA technology, the expression of clathrin (CLR) in EBL cells was knocked down to further verify the role of CLR in the invasion process of M. bovis. The results showed that the optimal conditions for M. bovis to invade non-phagocytic cells were a multiplicity of infection (MOI) of 1000 and an optimal invasion time of 4 h. All three strains of M. bovis have the ability to invade the four types of non-phagocytic cells, yet their invasion abilities vary significantly. Observations from transmission electron microscopy further confirmed that at 120 min post-infection, PG45 had successfully invaded EBL cells and was present within endocytic vesicles. It is noteworthy that almost all PG45 successfully escaped from the endocytic vesicles after 240 min of infection had passed. Through chemical inhibition experiments and CLR protein knockdown experiments, it was found that when the CDE and lipid raft pathways were blocked or CLR protein expression was reduced, the invasion rates of PG45, 07801, and 08M in MDBK, EBL, EBTr, and BT cells were significantly decreased (p < 0.05). The above results indicate that M. bovis can invade all types of non-phagocytic cells through endocytic pathways involving CDE (clathrin-dependent endocytosis) or lipid raft-mediated endocytosis, and possesses the ability to escape from phagosomes. Full article

Halophytes, such as Salicornia species, are promising new foods and are consumed for their pleasant salty taste and nutritional value. Since Salicornia is perishable, modified atmospheric packaging (MAP) can be a useful tool, in combination with proper temperature, to halt further quality degradation in this type of product. The purpose of this study was to investigate the effect of MAP, with or without refrigeration, to extend the shelf life of glasswort (Salicornia europaea L.) grown hydroponically (floating raft system) in a greenhouse with a nutrient solution containing 0 g/L (C) or 12.5 g/L of NaCl (T). The dry matter content, weight loss, respiration rate, biochemical composition, color, antioxidant capacity, and sensorial attributes were determined in shoots after harvest and during storage in plastic bags filled with technical air or with MAP at 4 or 20 °C for 120 h. At harvest, plants supplied with salt-enriched solution (T) showed a significant improvement in nutritional value and sensory profile. Storage in air at room temperature (20 °C) accelerated weight loss and diminished color stability, particularly in non-salinity samples (C), while MAP extended the shelf life of all the samples regardless of the storage temperature adopted. Optimal storage conditions were observed when MAP was combined with refrigeration, which allowed to effectively preserve shoots sensory acceptability for a period of about seven days. Future research could further explore the long-term effects on the nutritional value and sensory quality of S. europaea under various combinations of MAP and different storage temperatures ranging between 4 °C and 20 °C. Full article