Supersoft Elastic Bottlebrush Microspheres with Stimuli-Responsive Color-Changing Properties in Brine.

Solvent-free supersoft elastomer is highly desirable for building photonic structures with significant stimuli-responsive color changes. We report supersoft elastic porous microspheres with vivid structural colors obtained via self-assembly of amphiphilic bottlebrush block copolymers at the water/oil interface templated by ordered water-in-oil-in-water double emulsions. The porous structure is composed of cross-linked bottlebrush polydimethylsiloxane (PDMS) as the supersoft elastic skeleton and bottlebrush poly(ethylene oxide) (PEO) as the internal responsive layer. The obtained microspheres show large reversible volume changes through well-controlled dehydration or hydration of PEO in response to salt ions in an aqueous environment. As a result, full-spectrum colors are obtained dependent on different salt concentrations. In-situ observation of color reflection of a microsphere indicates a gradual structural transition from the outside to the inside corresponding to migration of water molecules and salt ions. Moreover, rod-like bottlebrush PEO exhibits an anion-induced salting-out behavior different from that of random coil polymers. The significantly responsive behaviors of bottlebrush block copolymer (BBCP) assemblies in the presence of salt ions primarily rely on the supersoft elastic skeleton of the porous structure, providing a facile route to the creation of stimuli-responsive photonic materials by low-cost self-assembly methods.

A minimization principle for transition paths of maximum flux for collective variables.

Considered is the construction of transition paths of conformational changes for proteins and other macromolecules, using methods that do not require the generation of dynamics trajectories. Special attention is given to the use of a reduced set of collective variables for describing such paths. A favored way to define transition paths is to seek channels through the transition state having cross sections with a high reactive flux (density of last hitting points of reactive trajectories). Given here is a formula for reactive flux that is independent of the parameterization of "collective variable space." This formula is needed for the principal curve of the reactive flux (as in the revised finite temperature string method) and for the maximum flux transition (MaxFlux) path. Additionally, a resistance functional is derived for narrow tubes, which when minimized yields a MaxFlux path. A strategy for minimization is outlined in the spirit of the string method. Finally, alternative approaches based on determining trajectories of high probability are considered, and it is observed that they yield paths that depend on the parameterization of collective variable space, except in the case of zero temperature, where such a path coincides with a MaxFlux path.

Interplay between insecticide-treated bed-nets and mosquito demography: implications for malaria control.

Although malaria prevalence has witnessed a significant reduction within the past decade, malaria still constitutes a major health and economic problem, especially to low-income countries. Insecticide-treated nets (ITNs) remain one of the primary measures for preventing the malignant disease. Unfortunately, the success of ITN campaigns is hampered by improper use and natural decay in ITN-efficacy over time. Many models aimed at studying malaria transmission and control fail to account for this decay, as well as mosquito demography and feeding preferences exhibited by mosquitoes towards humans. Omitting these factors can misrepresent disease risk, while understanding their effects on malaria dynamics can inform control policy. We present a model for malaria dynamics that incorporates these factors, and a systematic analysis, including stability and sensitivity analyses of the model under different conditions. The model with constant ITN-efficacy exhibits a backward bifurcation emphasizing the need for sustained control measures until the basic reproduction number, R0, drops below a critical value at which control is feasible. The infectious and partially immune human populations and R0 are highly sensitive to the probability that a mosquito feeds successfully on a human, ITN coverage and the maximum biting rate of mosquitoes, irrespective of whether ITN-efficacy is constant or declines over time. This implies that ITNs play an important role in disease control. When ITN-efficacy wanes over time, we identify disease risks and corresponding ITN coverage, as well as feeding preference levels for which the disease can be controlled or eradicated. Our study leads to important insights that could assist in the design and implementation of better malaria control strategies. We conclude that ITNs that can retain their effectiveness for longer periods will be more appropriate in the fight against malaria and that making more ITNs available to highly endemic regions is necessary for malaria containment.

[Toxoplasma gondii Rhoptry Protein 17 Inhibits the Apoptosis of Mouse Macrophages via Activation of Activator Protein 1 Signaling].

Objective: To investigate the effect of Toxoplasma gondii rhoptry protein 17（ROP17） on γ-interferon (IFN-γ)-induced apoptosis of mouse J774A.1 monocyte macrophages. Methods: The J774A.1 cells were transfected with recombinant plasmid p3×Flag-CMV-14/TgROP17 or empty plasmid p3×Flag-CMV-14. After addition of IFN-γ, flow cytometry and Western blotting were performed to detect apoptosis and the protein levels of phosphorylated c-Jun and apoptosis-related proteins cleaved Caspase-3, Bcl-2, Bcl-xL and Bcl-3. The p3×Flag-CMV-14/TgROP17 plasmid and c-Jun shRNA were co-transfected into J774A.1 cells, after which IFN-γ was added to induce cell apoptosis. The levels of cleaved Caspase-3 and Bcl-3 were analyzed using Western blotting. Results: Flow cytometry showed that the apoptosis rate of cells overexpressing ROP17［（3.73±0.51）%ï¼½ was significantly lower than that of the control cells［（7.78±1.10）%, P<0.05ï¼½. Western blotting showed significant differences in protein levels of phosphorylated c-Jun（0.196±0.028 vs. 0.075±0.010）, Bcl-3（0.461±0.063 vs. 0.108±0.013） and cleaved Caspase 3（0.015±0.004 vs. 0.174±0.026） between the cells overexpressing ROP17 and control cells （all P<0.05）. However, the levels of Bcl-2 and Bcl-xL were not significantly different between the cells overexpressing ROP17 and the control. When the expression of c-Jun and phosphorylation of c-Jun were inhibited by c-Jun shRNA, the relative level of cleaved Caspase 3 in the RNA interferenced cells and control cells was 0.147±0.024 and 0.087±0.010, respectively (P<0.05), and the relative level of Bcl-3 was 0.085±0.010 and 0.162±0.011, respectively (P<0.05). Conclusion: The anti-apoptosis effect of ROP17 is dependent on the phosphorylation of c-Jun and the expression of Bcl-3.

Quantifying the impact of decay in bed-net efficacy on malaria transmission.

Insecticide-treated nets (ITNs) are at the forefront of malaria control programs and even though the percentage of households in sub-Saharan Africa that owned nets increased from 3% in 2000 to 53% in 2012, many children continue to die from malaria. The potential impact of ITNs on reducing malaria transmission is limited due to inconsistent or improper use, as well as physical decay in effectiveness. Most mathematical models for malaria transmission have assumed a fixed effectiveness rate for bed-nets, which can overestimate the impact of nets on malaria control. We develop a model for malaria spread that captures the decrease in ITN effectiveness due to physical and chemical decay, as well as human behavior as a function of time. We perform uncertainty and sensitivity analyses to identify and rank parameters that play a critical role in malaria transmission. These analyses show that the basic reproduction number R0, and the infectious human population are most sensitive to bed-net coverage and the biting rate of mosquitoes. Our results show the existence of a backward bifurcation for the case in which ITN efficacy is constant over time, which occurs for some range of parameters and is characterized by high malaria mortality in humans. This result implies that bringing R0 to less than one is not enough for malaria elimination but rather additional efforts will be necessary to control the disease. For the case in which ITN efficacy decays over time, we determine coverage levels required to control malaria for different ITN efficacies and demonstrate that ITNs with longer useful lifespans perform better in malaria control. We conclude that malaria control programs should focus on increasing bed-net coverage, which can be achieved by enhancing malaria education and increasing bed-net distribution in malaria endemic regions.

Modeling the synergistic interplay between malaria dynamics and economic growth.

The mosquito-borne disease (malaria) imposes significant challenges on human health, healthcare systems, and economic growth/productivity in many countries. This study develops and analyzes a model to understand the interplay between malaria dynamics, economic growth, and transient events. It uncovers varied effects of malaria and economic parameters on model outcomes, highlighting the interdependence of the reproduction number (R0) on both malaria and economic factors, and a reciprocal relationship where malaria diminishes economic productivity, while higher economic output is associated with reduced malaria prevalence. This emphasizes the intricate interplay between malaria dynamics and socio-economic factors. The study offers insights into malaria control and underscores the significance of optimizing external aid allocation, especially favoring an even distribution strategy, with the most significant reduction observed in an equal monthly distribution strategy compared to longer distribution intervals. Furthermore, the study shows that controlling malaria in high mosquito biting areas with limited aid, low technology, inadequate treatment, or low economic investment is challenging. The model exhibits a backward bifurcation implying that sustainability of control and mitigation measures is essential even when R0 is slightly less than one. Additionally, there is a parameter regime for which long transients are feasible. Long transients are critical for predicting the behavior of dynamic systems and identifying factors influencing transitions; they reveal reservoirs of infection, vital for disease control. Policy recommendations for effective malaria control from the study include prioritizing sustained control measures, optimizing external aid allocation, and reducing mosquito biting.

The impact of bed-net use on malaria prevalence.

Malaria infection continues to be a major problem in many parts of the world including the Americas, Asia, and Africa. Insecticide-treated bed-nets have shown to reduce malaria cases by 50%; however, improper handling and human behavior can diminish their effectiveness. We formulate and analyze a mathematical model that considers the transmission dynamics of malaria infection in mosquito and human populations and investigate the impact of bed-nets on its control. The effective reproduction number is derived and existence of backward bifurcation is presented. The backward bifurcation implies that the reduction of R below unity alone is not enough to eradicate malaria, except when the initial cases of infection in both populations are small. Our analysis demonstrate that bed-net usage has a positive impact in reducing the reproduction number R. The results show that if 75% of the population were to use bed-nets, malaria could be eliminated. We conclude that more data on the impact of human and mosquito behavior on malaria spread is needed to develop more realistic models and better predictions.

PTPN1 is a prognostic biomarker related to cancer immunity and drug sensitivity: from pan-cancer analysis to validation in breast cancer.

Background: Protein tyrosine phosphatase non-receptor type 1 (PTPN1), a member of the protein tyrosine phosphatase superfamily, has been identified as an oncogene and therapeutic target in various cancers. However, its precise role in determining the prognosis of human cancer and immunological responses remains elusive. This study investigated the relationship between PTPN1 expression and clinical outcomes, immune infiltration, and drug sensitivity in human cancers, which will improve understanding regarding its prognostic value and immunological role in pan-cancer. Methods: The PTPN1 expression profile was obtained from The Cancer Genome Atlas and Cancer Cell Line Encyclopedia databases. Kaplan-Meier, univariate Cox regression, and time-dependent receiver operating characteristic curve analyses were utilized to clarify the relationship between PTPN1 expression and the prognosis of pan-cancer patients. The relationships between PTPN1 expression and the presence of tumor-infiltrated immune cells were analyzed using Estimation of Stromal and Immune cells in Malignant Tumor tissues using Expression data and Tumor Immune Estimation Resource. The cell counting kit-8 (CCK-8) assay was performed to examine the effects of PTPN1 level on the sensitivity of breast cancer cells to paclitaxel. Immunohistochemistry and immunoblotting were used to investigate the relationship between PTPN1 expression, immune cell infiltration, and immune checkpoint gene expression in human breast cancer tissues and a mouse xenograft model. Results: The pan-cancer analysis revealed that PTPN1 was frequently up-regulated in various cancers. High PTPN1 expression was associated with poor prognosis in most cancers. Furthermore, PTPN1 expression correlated highly with the presence of tumor-infiltrating immune cells and the expression of immune checkpoint pathway marker genes in different cancers. Furthermore, PTPN1 significantly predicted the prognosis for patients undergoing immunotherapy. The results of the CCK-8 viability assay revealed that PTPN1 knockdown increased the sensitivity of MDA-MB-231 and MCF-7 cells to paclitaxel. Finally, our results demonstrated that PTPN1 was associated with immune infiltration and immune checkpoint gene expression in breast cancer. Conclusion: PTPN1 was overexpressed in multiple cancer types and correlated with the clinical outcome and tumor immunity, suggesting it could be a valuable potential prognostic and immunological biomarker for pan-cancer.

A Novel Inflammatory Response-Related Gene Signature Predicts Immune Status and Prognosis of Breast Cancer.

Purpose: Breast cancer is the most common type of cancer and the leading cause of cancer-related death in women worldwide. In this study, we aimed to construct an inflammatory response-related gene model for predicting the immune status and prognosis of breast cancer patients. Methods: We obtained the inflammatory response-related genes from the Molecular Signatures Database. Furthermore, we used univariate Cox regression analysis, the least absolute shrinkage and selection operator (LASSO) regression analysis, and multivariate Cox regression to construct an inflammatory response-related gene signature (IRGS) model based on dataset obtained from The Cancer Genome Atlas (TCGA). Patients were consequently categorized into high-risk and low-risk groups. Kaplan-Meier analysis was used to compare the overall survival (OS) of high-risk and low-risk groups. Following that, we validated the model using a dataset (GSE96058) acquired from Gene Expression Omnibus (GEO) database. Univariate and multivariate Cox analyses were used to determine the independent prognostic value of the IRGS in the TCGA and GSE96058 cohorts. A nomogram was constructed to predict the OS in the TCGA cohort. Further, we used Gene Set Enrichment Analysis (GSEA), CIBERSORT, and single-sample Gene Set Enrichment Analysis (ssGSEA) to evaluate the associations of IRGS with immune-associated pathways and immune infiltration. Finally, the relationship between the expression of the signature genes and drug sensitivity was conducted using Pearson correlation analysis. Results: We established an IRGS to stratify breast cancer patients into the low-risk and high-risk groups. In both the training and validation sets, patients in the high-risk group had significantly shorter OS than those in the low-risk group. The risk score was significantly correlated with the clinical characteristics and could be used as a tool to predict the prognosis of breast cancer. Moreover, we found that the IRGS risk score was an independent predictor of OS in breast cancer patients, and a nomogram model based on IRGS risk score and other clinical factors could effectively predict the prognosis of breast cancer patients. Furthermore, the IRGS risk score was correlated with immune characteristics and was inversely associated with the abundance of immune cell infiltration. Patients with a low IRGS risk score had higher expression levels of immune checkpoint genes, suggesting that IRGS can be used as a potential indicator for immunotherapy. Finally, we found that the expression levels of prognostic genes were significantly correlated with tumor cell sensitivity to chemotherapeutic drugs. Conclusion: Overall, these findings suggest that the IRGS can be used to predict the prognosis and immune status of breast cancer patients and provide new therapeutic targets for the treatment of these patients.

PTPRO-related CD8+ T-cell signatures predict prognosis and immunotherapy response in patients with breast cancer.

Background: Poor immunogenicity and extensive immunosuppressive T-cell infiltration in the tumor immune microenvironment (TIME) have been identified as potential barriers to immunotherapy success in "immune-cold" breast cancers. Thus, it is crucial to identify biomarkers that can predict immunotherapy efficacy. Protein tyrosine phosphatase receptor type O (PTPRO) regulates multiple kinases and pathways and has been implied to play a regulatory role in immune cell infiltration in various cancers. Methods: ESTIMATE and single-sample gene set enrichment analysis (ssGSEA) were performed to uncover the TIME landscape. The correlation analysis of PTPRO and immune infiltration was performed to characterize the immune features of PTPRO. Univariate and multivariate Cox analyses were applied to determine the prognostic value of various variables and construct the PTPRO-related CD8+ T-cell signatures (PTSs). The Kaplan-Meier curve and the receiver operating characteristic (ROC) curve were used to estimate the performance of PTS in assessing prognosis and immunotherapy response in multiple validation datasets. Results: High PTPRO expression was related to high infiltration levels of CD8+ T cells, as well as macrophages, activated dendritic cells (aDCs), tumor-infiltrating lymphocytes (TILs), and Th1 cells. Given the critical role of CD8+ T cells in the TIME, we focused on the impact of PTPRO expression on CD8+ T-cell infiltration. The prognostic PTS was then constructed using the TCGA training dataset. Further analysis showed that the PTS exhibited favorable prognostic performance in multiple validation datasets. Of note, the PTS could accurately predict the response to immune checkpoint inhibitors (ICIs). Conclusion: PTPRO significantly impacts CD8+ T-cell infiltration in breast cancer, suggesting a potential role of immunomodulation. PTPRO-based PTS provides a new immune cell paradigm for prognosis, which is valuable for immunotherapy decisions in cancer patients.

Chemerin/ChemR23 signaling axis is involved in the endothelial protection by K(ATP) channel opener iptakalim.

AIM: To elucidate the modulation of the chemerin/ChemR23 axis by iptakalim-induced opening of K(ATP) channels and to determine the role of the chemerin/ChemR23 axis in the iptakalim-mediated endothelial protection. METHODS: Cultured rat aortic endothelial cells (RAECs) were used. Chemerin secretion and ChemR23 protein expression were investigated using Western blot analysis. The gene expression level of ChemR23 was examined with RT-PCR. In addition, the release of nitric oxide (NO) was measured with a nitric oxide assay. RESULTS: Homocysteine, uric acid, high glucose, or oxidized low-density lipoprotein (ox-LDL) down-regulated the chemerin secretion and ChemR23 gene/protein expression in RAECs as a function of concentration and time, which was reversed by pretreatment with iptakalim (1-10 µmol/L). Moreover, these effects of iptakalim were abolished in the presence of the K(ATP) channel antagonist glibenclamide (1 µmol/L). Both iptakalim and recombinant chemerin restored the impaired NO production in RAECs induced by uric acid, and the effects were abolished by anti-ChemR23 antibodies. CONCLUSION: Iptakalim via opening K(ATP) channels enhanced the endothelial chemerin/ChemR23 axis and NO production, thus improving endothelial function.

The G1 phase optical reporter serves as a sensor of CDK4/6 inhibition in vivo.

Visualization of cell-cycle G1 phase for monitoring the early response of cell cycle specific drug remains challenging. In this study, we developed genetically engineered bioluminescent reporters by fusing full-length cyclin E to the C-terminal luciferase (named as CycE-Luc and CycE-Luc2). Next, HeLa cell line or an ER-positive breast cancer cell line MCF-7 was transfected with these reporters. In cellular assays, the bioluminescent signal of CycE-Luc and CycE-Luc2 was accumulated in the G1 phase and decreased after exiting from the G1 phase. The expression of CycE-Luc and CycE-Luc2 fusion protein was regulated in a cell cycle-dependent manner, which was mediated by proteasome ubiquitination and degradation. Next, our in vitro and in vivo experiment confirmed that the cell cycle arrested by anti-cancer agents (palbociclib or 5-FU) was monitored quantitatively and dynamically by bioluminescent imaging of these reporters in a real-time and non-invasive manner. Thus, these optical reporters could reflect the G1 phase alternation of cell cycle, and might become a future clinically translatable approach for predicting and monitoring response to palbociclib in patients with ER-positive breast cancer.

Flow-Induced Crystallization in Polyethylene: Effect of Flow Time on Development of Shish-Kebab.

The flow-induced formation and relaxation of the representative oriented shish-kebab structure were studied with synchrotron small-angle X-ray scattering (SAXS) method. The flow duration was varied from 2 to 6 s at an identical strain rate to reveal the effect of flow time on stability and dimension of formed shish. It was found that the short flow time of 2 s was able to generate shish during flow, which, however, relaxed during the isothermal process after cessation of flow. An increase in flow time can improve the shish stability and the long flow time of 6 s can generate the stable shish that nucleate the growth of kebab lamellae. In addition, the quantitative analysis of SAXS results showed that with increasing flow time from 2 to 6 s, the shish length increased from 242 to 574 nm, while the shish diameter remained around 34 nm. This detailed information of the formed shish-kebab structure can be used to shed light on their evolution that occurred during flow from 2 to 6 s, where shish grew at a longitudinal speed of around 80 nm/s, and there was an improvement in the stability and nucleation capability for kebab lamellae.

A Mathematical model of Schistosoma mansoni in Biomphalaria glabrata with control strategies.

We describe and analyze a mathematical model for schistosomiasis in which infected snails are distinguished from susceptible through increased mortality and no reproduction. We based the model on the same derivation as Anderson and May (J. Anim. Ecol. 47:219-247, 1978), Feng and Milner (A New Mathematical Model of Schistosomiasis, Mathematical Models in Medical and Health Science, Nashville, TN, 1997. Innov. Appl. Math., Vanderbilt Univ. Press, Nashville, pp. 117-128, 1998), and May and Anderson (J. Anim. Ecol. 47:249-267, 1978), but used logistic growth both in human and snail hosts. We introduce a parameter r, the effective coverage of medical treatment/prevention to control the infection. We determine a reproductive number for the disease directly related to its persistence and extinction. Finally, we obtain a critical value for r that indicates the minimum treatment effort needed in order to clear out the disease from the population.

Electrochemical Behaviors of Methylene Blue on DNA Modified Electrode and Its Application to the Detection of PCR Product from NOS Sequence.

An electrochemical DNA biosensor for the detection of NOS gene sequences from genetically modified organisms (GMOs) is presented in this paper. Single-stranded DNA (ssDNA) was covalently attached through the carboxylate ester formed by the 3'- hydroxy end of the DNA with the carboxyl of a mercaptoacetic acid self-assembled monolayer-modified gold electrode using N-hydroxysuccinimide (NHS) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) as linkers. The electrochemical behavior of methylene blue (MB) on the ssDNA and dsDNA modified gold electrode were carefully studied. Compared with ssDNA/Au electrode, an increase of redox peak current of MB on dsDNA/Au electrode was found, which could be further used for monitoring the recognition of DNA hybridization. Based on this result, the polymerase chain reaction (PCR) product of the common inserts NOS terminator from real GMOs samples was detected successfully.

A mathematical model studying mosquito-stage transmission-blocking vaccines.

A compartmental deterministic model is proposed to evaluate the effectiveness of transmission-blocking vaccines of malaria, which targets at the parasite stage in the mosquito. The model is rigorously analyzed and numerical simulations are performed. The results and implications are discussed.

Bioluminescence imaging of DNA synthetic phase of cell cycle in living animals.

Bioluminescence reporter proteins have been widely used in the development of tools for monitoring biological events in living cells. Currently, some assays like flow cytometry analysis are available for studying DNA synthetic phase (S-phase) targeted anti-cancer drug activity in vitro; however, techniques for imaging of in vivo models remain limited. Cyclin A2 is known to promote S-phase entry in mammals. Its expression levels are low during G1-phase, but they increase at the onset of S-phase. Cyclin A2 is degraded during prometaphase by ubiquitin-dependent, proteasome-mediated proteolysis. In this study, we have developed a cyclin A2-luciferase (CYCA-Luc) fusion protein targeted for ubiquitin-proteasome dependent degradation, and have evaluated its utility in screening S-phase targeted anti-cancer drugs. Similar to endogenous cyclin A2, CYCA-Luc accumulates during S-phase and is degraded during G2/M-phase. Using Cdc20 siRNA we have demonstrated that Cdc20 can mediate CYCA-Luc degradation. Moreover, using noninvasive bioluminescent imaging, we demonstrated accumulation of CYCA-Luc in response to 10-hydroxycamptothecin (HCPT), an S-phase targeted anti-cancer drug, in human tumor cells in vivo and in vitro. Our results indicate that a CYCA-Luc fusion reporter system can be used to monitor S-phase of cell cycle, and evaluate pharmacological activity of anti-cancer drug HCPT in real time in vitro and in vivo, and is likely to provide an important tool for screening such drugs.

Identification of oligomer proanthocyanidins (F2) isolated from grape seeds as a formyl peptide receptor 1 partial agonist.

Formyl peptide receptor 1 (FPR1) plays an important role in the rapid progression of glioblastoma and has been considered as a molecular target for the treatment. Previously, we have shown that oligomer proanthocyanidins (F2, degree of polymerization 2-15), isolated from grape seeds, inhibited FPR1-mediated chemotaxis of U-87 glioblastoma cells. In the present study, we investigated the capacity of F2 to interact with FPR1. The cross attenuation of chemotaxis revealed that F2 shared FPR1 with formyl-methionyl-leucyl-phenylalanine (fMLF), which is a prototype agonist of FPR1. F2 was chemotactic for U-87 cells, and the chemotactic response was abolished when FPR1 gene was silenced or FPR1 was competitively occupied. We further show that F2 specifically blocked the binding of fluorescent agonist to FPR1. Interestingly, F2 exhibited the characteristic of a partial agonist for FPR1, as shown by its capacity to activate FPR1-mediated PI3K-PKC-MAPK pathways. Meanwhile, F2 also attenuated fMLF-triggered MAPK activation, suggesting that F2 could antagonize the effect of an agonist. Furthermore, F2 abolished the invasion of U-87 cells induced by fMLF. Thus, we have identified F2 as a novel, partial agonist for FPR1, which may be useful for glioblastoma therapy.

Stimulation of non-neuronal muscarinic receptors enhances chemerin/ChemR23 system in dysfunctional endothelial cells.

AIMS: Endothelial cells play a pivotal role in vascular intimal inflammation during cardiovascular diseases. The chemerin/ChemR23 system in endothelial cells is one of physiological mechanisms that regulate inflammatory responses. Our previous studies indicated that stimulation of non-neuronal muscarinic receptor (NNMR) improved endothelial dysfunction. However, the relationship between the chemerin/ChemR23 signaling axis and NNMR in endothelial cell is poorly understood. Here, we first investigated whether the modulation of chemerin/ChemR23 signaling axis is involved in NNMR-mediated endothelial protection. MAIN METHODS: Cultured rat aortic endothelial cells (RAECs) were used. The ChemR23 protein expression and chemerin secretion were measured using Western blot analysis. The gene expression level of ChemR23 was examined with reverse transcriptase PCR (RT-PCR). The production of nitric oxide (NO) was determined by a nitrate reductase assay kit. KEY FINDINGS: A sharp decline of chemerin secretion and ChemR23 protein/gene expression was observed in RAECs after exposed to homocysteine at concentration of 0.5 mmol/L. Arecoline (10 µmol/L) pretreatment increased ChemR23 protein expression as well as mRNA expression, and enhanced the secretion of chemerin. Arecoline could also reverse the decreased ChemR23 mRNA expression induced by uric acid, high glucose, or oxidized low-density lipoprotein. Furthermore, the modulation of arecoline on chemerin/ChemR23 signaling axis was absolutely abolished in the presence of the nonselective muscarinic receptors antagonist atropine 1 µmol/L. Additionally, arecoline improved endothelial dysfunction by increasing the reduced NO production induced by uric acid, which was blocked by anti-ChemR23 antibody. SIGNIFICANCE: The chemerin/ChemR23 signaling axis participates in NNMR-mediated protection against endothelial dysfunction in cardiovascular system.

αC helix as a switch in the conformational transition of Src/CDK-like kinase domains.

One mechanism of regulating the catalytic activity of protein kinases is through conformational transitions. Despite great diversity in the structural changes involved in the transitions, a certain set of changes within the kinase domain (KD) has been observed for many kinases including Src and CDK2. We investigated this conformational transition computationally to identify the topological features that are energetically critical to the transition. Results from both molecular dynamics sampling and transition path optimization highlight the displacement of the αC helix as the major energy barrier, mediating the switch of the KD between the active and down-regulated states. The critical role of the αC helix is noteworthy by providing a rationale for a number of activation and deactivation mechanisms known to occur in cells. We find that kinases with the αC helix displacement exist throughout the kinome, suggesting that this feature may have emerged early in evolution.