Computational modelling of dynamic cAMP responses to GPCR agonists for exploration of GLP-1R ligand effects in pancreatic ß-cells and neurons.

The glucagon-like peptide-1 receptor (GLP-1R) is a class B G protein-coupled receptor (GPCR) which plays important physiological roles in insulin release and promoting fullness. GLP-1R agonists initiate cellular responses by cyclic AMP (cAMP) pathway signal transduction. Understanding of the potential of GLP-1R agonists in the treatment of type 2 diabetes may be advanced by considering the cAMP dynamics for agonists at GLP-1R in both pancreatic ß-cells (important in insulin release) and neurons (important in appetite regulation). Receptor desensitisation in the cAMP pathway is known to be an important regulatory mechanism, with different ligands differentially promoting G protein activation and desensitisation. Here, we use mathematical modelling to quantify and understand experimentally obtained cAMP timecourses for two GLP-1R agonists, exendin-F1 (ExF1) and exendin-D3 (ExD3), which give markedly different signals in ß-cells and neurons. We formulate an ordinary differential equation (ODE) model for the dynamics of cAMP signalling in response to G protein-coupled receptor (GPCR) ligands, encompassing ligand binding, receptor activation, G protein activation, desensitisation and second messenger generation. We validate our model initially by fitting to timecourse data for HEK293 cells, then proceed to parameterise the model for ß-cells and neurons. Through numerical simulation and sensitivity studies, our analysis adds support to the hypothesis that ExF1 offers more potential glucose regulation benefit than ExD3 over long timescales via signalling in pancreatic ß-cells, but that there is little difference between the two ligands in the potential appetite suppression effects offered via long-time signalling in neurons on the same timescales.

Ultrafast Metal-Free Microsupercapacitor Arrays Directly Store Instantaneous High-Voltage Electricity from Mechanical Energy Harvesters.

Harvesting renewable mechanical energy is envisioned as a promising and sustainable way for power generation. Many recent mechanical energy harvesters are able to produce instantaneous (pulsed) electricity with a high peak voltage of over 100 V. However, directly storing such irregular high-voltage pulse electricity remains a great challenge. The use of extra power management components can boost storage efficiency but increase system complexity. Here utilizing the conducting polymer PEDOT:PSS, high-rate metal-free micro-supercapacitor (MSC) arrays are successfully fabricated for direct high-efficiency storage of high-voltage pulse electricity. Within an area of 2.4 × 3.4 cm2 on various paper substrates, large-scale MSC arrays (comprising up to 100 cells) can be printed to deliver a working voltage window of 160 V at an ultrahigh scan rate up to 30 V s-1 . The ultrahigh rate capability enables the MSC arrays to quickly capture and efficiently store the high-voltage (≈150 V) pulse electricity produced by a droplet-based electricity generator at a high efficiency of 62%, significantly higher than that (<2%) of the batteries or capacitors demonstrated in the literature. Moreover, the compact and metal-free features make these MSC arrays excellent candidates for sustainable high-performance energy storage in self-charging power systems.

Butyrate as a promising therapeutic target in cancer: From pathogenesis to clinic (Review).

Cancer is one of the leading causes of mortality worldwide. The etiology of cancer has not been fully elucidated yet, and further enhancements are necessary to optimize therapeutic efficacy. Butyrate, a short­chain fatty acid, is generated through gut microbial fermentation of dietary fiber. Studies have unveiled the relevance of butyrate in malignant neoplasms, and a comprehensive understanding of its role in cancer is imperative for realizing its full potential in oncological treatment. Its full antineoplastic effects via the activation of G protein­coupled receptors and the inhibition of histone deacetylases have been also confirmed. However, the underlying mechanistic details remain unclear. The present study aimed to review the involvement of butyrate in carcinogenesis and its molecular mechanisms, with a particular emphasis on its association with the efficacy of tumor immunotherapy, as well as discussing relevant clinical studies on butyrate as a therapeutic target for neoplastic diseases to provide new insights into cancer treatment.

Abolishing ß-arrestin recruitment is necessary for the full metabolic benefits of G protein-biased glucagon-like peptide-1 receptor agonists.

AIM: Earlier studies have shown that peptide glucagon-like peptide-1 receptor (GLP-1R) agonists with reduced ß-arrestin recruitment show enhanced anti-hyperglycaemic efficacy through avoidance of GLP-1R desensitization. However, the ligand modifications needed to decrease ß-arrestin recruitment usually also reduces GLP-1R affinity, therefore higher doses are needed. Here we aimed to develop new, long-acting, G protein-biased GLP-1R agonists with acute signalling potency comparable with semaglutide, to provide insights into specific experimental and therapeutic scenarios. MATERIALS AND METHODS: New GLP-1R agonist peptides were assessed using a variety of in vitro and in vivo assays. RESULTS: First, we show that very substantial reductions in ß-arrestin recruitment efficacy are required to realize fully the benefits of GLP-1R agonism on blood glucose lowering in mice, with more moderate reductions being less effective. Secondly, our lead compound (SRB107) performs substantially better than semaglutide for effects on blood glucose and weight loss, which may be jointly attributable to its biased agonist action and protracted pharmacokinetics. Thirdly, we show that biased agonist-specific GLP-1R internalization profiles occur at clinically relevant pharmacological concentrations. Finally, we show that SRB107 cAMP signalling is differentially modulated by single and double GLP1R coding variants seen in human populations, with implications for GLP-1R agonist pharmacogenomics. CONCLUSIONS: Completely abolishing ß-arrestin recruitment improves the anti-hyperglycaemic effects of GLP-1R agonists in mice.

GWAS of random glucose in 476,326 individuals provide insights into diabetes pathophysiology, complications and treatment stratification.

Conventional measurements of fasting and postprandial blood glucose levels investigated in genome-wide association studies (GWAS) cannot capture the effects of DNA variability on 'around the clock' glucoregulatory processes. Here we show that GWAS meta-analysis of glucose measurements under nonstandardized conditions (random glucose (RG)) in 476,326 individuals of diverse ancestries and without diabetes enables locus discovery and innovative pathophysiological observations. We discovered 120 RG loci represented by 150 distinct signals, including 13 with sex-dimorphic effects, two cross-ancestry and seven rare frequency signals. Of these, 44 loci are new for glycemic traits. Regulatory, glycosylation and metagenomic annotations highlight ileum and colon tissues, indicating an underappreciated role of the gastrointestinal tract in controlling blood glucose. Functional follow-up and molecular dynamics simulations of lower frequency coding variants in glucagon-like peptide-1 receptor (GLP1R), a type 2 diabetes treatment target, reveal that optimal selection of GLP-1R agonist therapy will benefit from tailored genetic stratification. We also provide evidence from Mendelian randomization that lung function is modulated by blood glucose and that pulmonary dysfunction is a diabetes complication. Our investigation yields new insights into the biology of glucose regulation, diabetes complications and pathways for treatment stratification.

Enhanced Endosomal Signaling and Desensitization of GLP-1R vs GIPR in Pancreatic Beta Cells.

The incretin receptors, glucagon-like peptide-1 receptor (GLP-1R) and glucose-dependent insulinotropic polypeptide receptor (GIPR), are prime therapeutic targets for the treatment of type 2 diabetes (T2D) and obesity. They are expressed in pancreatic beta cells where they potentiate insulin release in response to food intake. Despite GIP being the main incretin in healthy individuals, GLP-1R has been favored as a therapeutic target due to blunted GIPR responses in T2D patients and conflicting effects of GIPR agonists and antagonists in improving glucose tolerance and preventing weight gain. There is, however, a recently renewed interest in GIPR biology, following the realization that GIPR responses can be restored after an initial period of blood glucose normalization and the recent development of dual GLP-1R/GIPR agonists with superior capacity for controlling blood glucose levels and weight. The importance of GLP-1R trafficking and subcellular signaling in the control of receptor outputs is well established, but little is known about the pattern of spatiotemporal signaling from the GIPR in beta cells. Here, we have directly compared surface expression, trafficking, and signaling characteristics of both incretin receptors in pancreatic beta cells to identify potential differences that might underlie distinct pharmacological responses associated with each receptor. Our results indicate increased cell surface levels, internalization, degradation, and endosomal vs plasma membrane activity for the GLP-1R, while the GIPR is instead associated with increased plasma membrane recycling, reduced desensitization, and enhanced downstream signal amplification. These differences might have potential implications for the capacity of each incretin receptor to control beta cell function.

Prediction of bimetal embedded in two-dimensional materials for CO2 reduction electrocatalysis with a new integrated descriptor.

CO2 reduction catalysis plays an important role in the process of converting harmful exhaust gas into useful fuels. However, the product complexity and the difficult hydrogenation in critical steps make it difficult to find a suitable catalyst for CO2 reduction. In this work, we report homo/hetero bimetal embedded in two-dimensional materials for electrocatalysis and discovered a new descriptor. We chose ß12-borophene accommodating two transition metal atoms for efficient CO2RR as a model system. We found that MnCo and VV systems are promising for CO2 reduction with good stability and high selectivity over HER. Through least absolute shrinkage and selection operator (LASSO) regression, we discovered a new integrated descriptor containing the spin moment of the metals and the descriptor is linked with the performance of the first step of CO2 hydrogenation. The MnCo system could catalyze a C1 process with low free energy change of the rate determining step. The VV system could also conduct the C2 process with low free energy change of the rate determining step. Bader charge analysis shows the ability of the borophene substrate to provide or hold electrons. This work demonstrates homonuclear and heteronuclear biatomic catalysts with high activity for CO2RR.

Evaluation of efficacy- versus affinity-driven agonism with biased GLP-1R ligands P5 and exendin-F1.

The glucagon-like peptide-1 receptor (GLP-1R) is an important regulator of glucose homeostasis and has been successfully targeted for the treatment of type 2 diabetes. Recently described biased GLP-1R agonists with selective reductions in ß-arrestin versus G protein coupling show improved metabolic actions in vivo. However, two prototypical G protein-favouring GLP-1R agonists, P5 and exendin-F1, are reported to show divergent effects on insulin secretion. In this study we aimed to resolve this discrepancy by performing a side-by-side characterisation of these two ligands across a variety of in vitro and in vivo assays. Exendin-F1 showed reduced acute efficacy versus P5 for several readouts, including recruitment of mini-G proteins, G protein-coupled receptor kinases (GRKs) and ß-arrestin-2. Maximal responses were also lower for both GLP-1R internalisation and the presence of active GLP-1R-mini-Gs complexes in early endosomes with exendin-F1 treatment. In contrast, prolonged insulin secretion in vitro and sustained anti-hyperglycaemic efficacy in mice were both greater with exendin-F1 than with P5. We conclude that the particularly low acute efficacy of exendin-F1 and associated reductions in GLP-1R downregulation appear to be more important than preservation of endosomal signalling to allow sustained insulin secretion responses. This has implications for the ongoing development of affinity- versus efficacy-driven biased GLP-1R agonists as treatments for metabolic disease.

Ligand-Specific Factors Influencing GLP-1 Receptor Post-Endocytic Trafficking and Degradation in Pancreatic Beta Cells.

The glucagon-like peptide-1 receptor (GLP-1R) is an important regulator of blood glucose homeostasis. Ligand-specific differences in membrane trafficking of the GLP-1R influence its signalling properties and therapeutic potential in type 2 diabetes. Here, we have evaluated how different factors combine to control the post-endocytic trafficking of GLP-1R to recycling versus degradative pathways. Experiments were performed in primary islet cells, INS-1 832/3 clonal beta cells and HEK293 cells, using biorthogonal labelling of GLP-1R to determine its localisation and degradation after treatment with GLP-1, exendin-4 and several further GLP-1R agonist peptides. We also characterised the effect of a rare GLP1R coding variant, T149M, and the role of endosomal peptidase endothelin-converting enzyme-1 (ECE-1), in GLP1R trafficking. Our data reveal how treatment with GLP-1 versus exendin-4 is associated with preferential GLP-1R targeting towards a recycling pathway. GLP-1, but not exendin-4, is a substrate for ECE-1, and the resultant propensity to intra-endosomal degradation, in conjunction with differences in binding affinity, contributes to alterations in GLP-1R trafficking behaviours and degradation. The T149M GLP-1R variant shows reduced signalling and internalisation responses, which is likely to be due to disruption of the cytoplasmic region that couples to intracellular effectors. These observations provide insights into how ligand- and genotype-specific factors can influence GLP-1R trafficking.

The Influence of Peptide Context on Signaling and Trafficking of Glucagon-like Peptide-1 Receptor Biased Agonists.

Signal bias and membrane trafficking have recently emerged as important considerations in the therapeutic targeting of the glucagon-like peptide-1 receptor (GLP-1R) in type 2 diabetes and obesity. In the present study, we have evaluated a peptide series with varying sequence homology between native GLP-1 and exendin-4, the archetypal ligands on which approved GLP-1R agonists are based. We find notable differences in agonist-mediated cyclic AMP signaling, recruitment of ß-arrestins, endocytosis, and recycling, dependent both on the introduction of a His → Phe switch at position 1 and the specific midpeptide helical regions and C-termini of the two agonists. These observations were linked to insulin secretion in a beta cell model and provide insights into how ligand factors influence GLP-1R function at the cellular level.

Design of a Graphene Nitrene Two-Dimensional Catalyst Heterostructure Providing a Well-Defined Site Accommodating One to Three Metals, with Application to CO2 Reduction Electrocatalysis for the Two-Metal Case.

Recently, the reduction of CO2 to fuels has been the subject of numerous studies, but the selectivity and activity remain inadequate. Progress has been made on single-site two-dimensional catalysts based on graphene coupled to a metal and nitrogen for the CO2 reduction reaction (CO2RR); however, the product is usually CO, and the metal-N environment remains ambiguous. We report a novel two-dimensional graphene nitrene heterostructure (grafiN6) providing well-defined active sites (N6) that can bind one to three metals for the CO2RR. We find that homobimetallic FeFe-grafiN6 could reduce CO2 to CH4 at -0.61 V and to CH3CH2OH at -0.68 V versus reversible hydrogen electrode, with high product selectivity. Moreover, the heteronuclear FeCu-grafiN6 system may be significantly less affected by hydrogen evolution reaction, while maintaining a low limiting potential (-0.68 V) for C1 and C2 mechanisms. Binding metals to one N6 site but not the other could promote efficient electron transport facilitating some reaction steps. This framework for single or multiple metal sites might also provide unique catalytic sites for other catalytic processes.

Predicted Optimal Bifunctional Electrocatalysts for the Hydrogen Evolution Reaction and the Oxygen Evolution Reaction Using Chalcogenide Heterostructures Based on Machine Learning Analysis of in Silico Quantum Mechanics Based High Throughput Screening.

Two-dimensional van der Waals heterostructure materials, particularly transition metal dichalcogenides (TMDC), have proved to be excellent photoabsorbers for solar radiation, but performance for such electrocatalysis processes as water splitting to form H2 and O2 is not adequate. We propose that dramatically improved performance may be achieved by combining two independent TMDC while optimizing such descriptors as rotational angle, bond length, distance between layers, and the ratio of the bandgaps of two component materials. In this paper we apply the least absolute shrinkage and selection operator (LASSO) process of artificial intelligence incorporating these descriptors together with quantum mechanics (density functional theory) to predict novel structures with predicted superior performance. Our predicted best system is MoTe2/WTe2 with a rotation of 300°, which is predicted to have an overpotential of 0.03 V for HER and 0.17 V for OER, dramatically improved over current electrocatalysts for water splitting.

Continual in-plane displacement measurement with temporal wavelet transform speckle pattern interferometry.

A heterodyne temporal speckle pattern interferometer that measures the in-plane displacement dynamically has been built. The object is displaced in its plane continuously and the frequency-modulated output signals with a carrier frequency are recorded by a CCD camera. The displacement information is extracted with wavelet transform technique. Preliminary experiments have been performed with such interferometer. The respective measurement results recovered from wavelet transform and Fourier transform are compared.

[The changes of hemodynamic parameters, pathology and c-kit mRNA expression in myocardium after acute myocardial infarction in rats].

This study was aimed to investigate the changes of hemodynamic parameters, pathology and c kit mRNA expression in myocardium after acute myocardial infarctionin (AMI) in rats, and to elucidate the relationship between these three kinds of changes. Sixty six adult male SD rats were randomly divided into normal group, Sham groups and ligation groups. The rat model of AMI was set up by ligating the left anterior descending artery. Hemodynamic parameters, pathological changes and c kit mRNA expression in myocardiam were examined. The results revealed that there were no statistically significant differences in hemodynamic parameters between normal group and Sham groups. Compared with the normal group, all ligation groups exhibited significantly decreased left ventricular systolic pressure (LVSP) and +/-dp/dtmax (P<0.01), and increased left ventricular end diastolic pressure (LVEDP, P<0.01). In the other ligation groups, compared with 6th hour group after ligation, there appeared striking increase of LVSP, LVEDP and +/-dp/dtmax (P<0.05). HE staining in myocardiam showed that there are necrosis and derangement at 24th hour group after ligation ,and a great number of inflammatory cells infiltration around the infarct zone at 3rd day group after ligation, and granulation tissue infiltrated into the infarct zone at 14th day group after ligation. In all five time points groups after ligation, the levels of c-kit mRNA expression were 0.99 fold, 1.06 fold, 1.46 fold, 1.91 fold and 2.67 fold, respectively, compared with Sham groups. The results suggest that cardiac stem cells in myocardium might contribute to the role of regenerating myocardium via self proliferation after acute myocardial infarction, but further investigation is still needed.

[Effects of ovariectomy and estradiol on microarchitecture and biomechanical properties in rat vertebrae].

This study was aimed at observing the effects of ovariectomy and estradiol on the microarchitecture of cancellous bone and exploring the influence of microarchitectural change on the biomechanical properties. Thirty 6-month-old unmated female SD rats were randomly divided into 3 groups (10 rat each): sham-operated control group (Sham), ovariectomized group (OVX)and Estradiol Benzoate treated group (EBT). All rats were housed in standard environmental conditions. Five months after operation, the rats were sacrificed. The biomechanical properties of the third lumbar vertebras (L3) were measured with compression testing in vitro. Micro-CT scanning was performed on the fourth lumbar vertebras (L4) in vitro. In comparison with the corresponding variables of Sham, the bone volume fraction (BV/TV) and the trabecular number (Tb. N) of OVX were reduced remarkably, and the trabecular separation (Tb. Sp) and the structural model index (SMI) of OVX were enhanced obviously. These facts implicated that the bone trabecular plate-like structure of OVX were decreased. BV/TV, Tb. N and the trabecular thickness (Tb. Th) of EBT were greater than those of OVX. Tb. Sp and SMI of EBT were much smaller than those of OVX. The results of mechanical test showed that the maximum forioe (Fmax), the maximum stress (sigmamax) and the elastic modeulus (E) of the lumbar vertebral cancellous bone of OVX were declined sharply, while the aforesaid biomechanical index of EBT was improved distinctly. The performance of three-dimensional micro-CT and the mechanical testing to assess microarchitecture of cancellous bone are useful for evaluating the state of osteoporosis and the antiosteoporotic effect of agents.

Effect of chitosan molecular weight and deacetylation degree on hemostasis.

Comparative studies have been carried out among solid-state chitosan soliquoid, chitosan acetic acid physiological saline solution, and carboxymethyl chitosan physiological saline solution to discover the hemostatic effect of molecular weight (M(w)) and deacetylation degree (DA) of chitosan. It was found that solid-state chitosan and chitosan acetic acid physiological saline solution performed different hemostatic mechanisms. When blood mixed with chitosan acetic acid physiological saline solution, the erythrocytes aggregated and were deformed. The DA, especially a low DA, in the chitosan acetic acid physiological saline solution, had a significant effect on the unusual aggregation and deformation of erythrocytes, compared with the effect of M(w) within a range between 10(5) and 10(6). However, this phenomenon could not be observed in solid-state chitosan soliquoid. Solid-state chitosan with a low DA absorbed more platelets and was more hemostatic. Carboxymethyl chitosan physiological saline solution had nothing to do with the aggregation and deformation of erythrocytes but caused local rouleau. The values of thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (APTT), and fibrinogen concentration (FIB) were measured after the blood was mixed with solid-state chitosan soliquoid, chitosan acetic acid physiological saline solution, and carboxymethyl chitosan physiological saline solution, separately. The results demonstrated that coagulation factors might not be activated by them.

[Research of the hemostasis effect of chitosan acetic acid solution].

The effect of chitosan with different molecular weight and deacetylation degree on blood hemostasis was tested. The experiments found evident alteration of red blood cell morphology and unusual coagglutination between erythrocytes in the anticoagulant blood which was treated by chitosan acetic acid solution. The red blood cells clot formation experiments showed that chitosan with low deacetylation degree (60%-70%) caused more red blood cells to assemble when compared versus chitosan with other deacetylation degrees. The effect of molecular weight between 10(5)-10(6) was not obvious. The thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (APTT), and concentration of fibrinogen (FIB) of blood treated by chitosan acetic acid solution were measured. The results proved that the hemostasis property of chitosan acetic acid solution was independent of the platelets and the normal "Cascade-like" coagulation pathway.