Associations of iron metabolism and inflammation with all-cause and cardiovascular mortality in a large NHANES community sample: Moderating and mediating effects.

BACKGROUND AND AIMS: This study aimed to assess the associations between serum iron concentration, C-reactive protein (CRP) concentration and the risk of all-cause mortality and cardiovascular mortality in the general population and to explore potential mediating and moderating effects. METHODS AND RESULTS: This study analyzed data from the National Health and Nutrition Examination Survey spanning the years 1999-2010, encompassing 23,634 participants. Cox proportional hazards regression models were employed to investigate the independent associations of serum iron and CRP with all-cause and cardiovascular mortality. Moderation and mediation analyses explored the moderating effect of CRP on the association between the serum iron concentration and all-cause and cardiovascular mortality, and the mediating role of the serum iron concentration in the association between the CRP concentration and all-cause and cardiovascular mortality. After multivariate adjustments in the Cox model, serum iron and CRP levels were independently correlated with both all-cause and cardiovascular mortality risk. Moderation analyses revealed a more pronounced correlation between the serum iron concentration and both all-cause and cardiovascular mortality in participants with higher CRP levels. Mediation analysis indicated that the serum iron concentration partly mediated the impact of CRP on the risk of all-cause mortality (13.79%) and cardiovascular mortality (24.12%). CONCLUSION: Serum iron and CRP are independently associated with all-cause and cardiovascular mortality. Moreover, the associations between serum iron concentrations and both all-cause and cardiovascular mortality are more pronounced in individuals with elevated CRP. Serum iron partially mediates the effect of CRP on all-cause and cardiovascular mortality.

A Coarse-Grained Interaction Model for Sodium Dominant Montmorillonite.

Montmorillonite is the main crystalline mineral present in bentonite. It is an absorbent, swelling material; the physical chemistry underlying its ability to absorb water and swell occurs at the nanoscale, governed by electrical double-layer interactions. In turn, absorption and swelling lead to important changes in the macroscopic transport properties of the clay. Mesoscale models can help us establish a link between these nanoscale processes and macroscale properties, notably by providing a detailed description of its pore network. Models on the scale of hundreds to thousands of nanometers are required, which cannot realistically be handled using traditional all-atom molecular dynamics simulations. This work presents a coarse-grained (CG) mesoscale model of sodium montmorillonite. In our model, montmorillonite platelets are represented by two types of particles: central nonhydrogen-bonded particles and edge hydrogen-bonding particles. The particle interactions are described by two-body potentials, which were optimized based on all-atom molecular dynamics simulations. Specifically, several potential mean force calculations involving dry and hydrated montmorillonite were performed, using the ClayFF potential to calculate interatomic forces. The CG model was validated by testing the scalability of the model, testing its ability to reproduce potentials of mean force reported elsewhere in the literature, and by comparing the calculated elastic properties of a system containing 1000 Na montmorillonite platelets to experimentally measured elastic properties of bentonite. The simulated elastic properties obtained using our mesoscale model agree with these experimental values.

Silver oxide model surface improves computational simulation of surface-enhanced Raman spectroscopy on silver nanoparticles.

Surface-enhanced Raman spectroscopy (SERS) coupled with density functional theory (DFT) computations can characterise the adsorption orientation of a molecule on a nanoparticle surface. When using DFT to simulate SERS on a silver surface, one typically employs an atom (Ag), ion (Ag+), or cluster (Agx or Agx+) as the model surface. Here, by examining the nucleobase 2,6-diaminopurine (2,6-DAP) and then generalising our strategy to three other molecules, we show that employing silver oxide (Ag2O) as the model surface can quantitatively improve the accuracy of simulated SERS.

High pressure chemistry of thioaldehydes: A first-principles molecular dynamics study.

First-principles molecular dynamics simulations are used to investigate the chemical behavior of bulk thioacetaldehyde (MeC(H)S) in response to changes in pressure, P. The simulations show that these molecules oligomerize in response to applied P. Oligomerization is initiated through C-S bond formation, with constrained dynamics simulations showing that the barrier to this reaction step is lowered significantly by applied P. Subsequent reactions involving the formation of additional C-S bonds or radical processes that lead to S-S and C-C bonds lengthen the oligomers. Oligomerization is terminated through proton transfer or the formation of rings. The mechanistic details of all reactions are examined. The results indicate that the P-induced reactivity of the MeC(H)S-based system differs significantly from that of analogous MeC(H)O-based systems, which have been reported previously. Comparison with the MeC(H)O study shows that replacing oxygen with sulfur significantly lowers the P required to initiate oligomerization (from 26 GPa to 5 GPa), increases the types of reactions in which systems of this type can take part, and increases the variety of products formed through these reactions. These differences can be explained in terms of the electronic structures of these systems, which may be useful for certain high P applications.

The mechanisms and therapeutic potential of clopidogrel in mitigating diabetic cardiomyopathy in db/db mice.

Clopidogrel has been shown to play a protective role against diabetic nephropathy. However, whether clopidogrel exerts a protective effect against diabetic cardiomyopathy (DCM) is unknown. Three-month-old male db/db mice were administered clopidogrel daily at doses of 5, 10, and 20 mg/kg by gavage for 5 months. Here, we showed that clopidogrel effectively attenuated diabetes-induced cardiac hypertrophy and cardiac dysfunction by inhibiting cardiac fibrosis, inflammatory responses, and oxidative stress damage in db/db mice. Diabetes-induced cardiac fibrosis was inhibited by clopidogrel treatment via blockade of the TGF-ß1/Smad3/P2RY12 pathway and inhibition of macrophage infiltration in db/db mice. The protective effects of clopidogrel against oxidative damage were mediated by the induction of the Nrf2 signaling pathway. Taken together, our findings provide strong evidence that clopidogrel is a promising effective agent for the treatment of DCM by alleviating diabetes-induced cardiac hypertrophy and dysfunction. P2RY12 might be an effective target for the treatment of DCM.

The causal effect of Helicobacter pylori infection on coronary heart disease is mediated by the body mass index: a Mendelian randomization study.

The association between Helicobacter pylori (H. pylori) infection and coronary heart disease (CHD) remains controversial, with an unclear causal link. This study employed bidirectional Mendelian randomization (MR) method, using H. pylori infection as the exposure, to investigate its causal relationship with CHD diagnosis, prognosis, and potential pathogenesis. H. pylori infection exhibited a causal association with body mass index (BMI) (ß = 0.022; 95% CI 0.008-0.036; p = 0.001). Conversely, there was no discernible connection between H. pylori infection and the diagnosis of CHD (OR = 0.991; 95% CI 0.904-1.078; p = 0.842; IEU database; OR = 1.049; 95% CI 0.980-1.118; p = 0.178; FinnGen database) or CHD prognosis (OR = 0.999; 95% CI 0.997-1.001; p = 0.391; IEU database; OR = 1.022; 95% CI 0.922-1.123; p = 0.663; FinnGen database). Reverse MR analysis showed no causal effect of CHD on H. pylori infection. Our findings further support that H. pylori infection exerts a causal effect on CHD incidence, mediated by BMI. Consequently, eradicating or preventing H. pylori infection may provide an indirect clinical benefit for patients with CHD.

Txnip promotes autophagic apoptosis in diabetic cardiomyopathy by upregulating FoxO1 and its acetylation.

Autophagy dysfunction and apoptosis exacerbate the risk of heart failure in patients with diabetic cardiomyopathy (DCM). However, the interactions between autophagy and apoptosis in DCM and their underlying mechanisms remain poorly understood. This study induced type 1 DCM in C57BL/6 mice via streptozotocin injection and exposed H9C2 cells to high glucose to investigate these mechanisms. The study revealed a significant elevation in autophagic vesicles and compromised autophagic flux, accompanied by pronounced myocardial cell apoptosis in the myocardium of diabetic mice. Long-term exposure to high glucose in H9C2 cells led to enhanced autophagosome formation and impaired autophagic flux, while inhibition of autophagy with 3-MA reduced cell apoptosis. Additionally, we observed an increase in Txnip expression in the myocardium of diabetic mice and in high glucose-treated H9C2 cells, which regulates autophagic apoptosis in high glucose-treated H9C2 cells. Furthermore, Txnip regulates autophagic apoptosis through the modulation of forkhead box-1 (FoxO1) expression and acetylation. Prolonged high glucose exposure resulted in increased levels of phosphorylated sirtuin 1 (SIRT1) and reduced SIRT1/FoxO1 interaction, changes that were ameliorated by Txnip knockdown. Txnip overexpression elevated FoxO1 levels, which could be suppressed by NAC and GSH. These findings revealed that Txnip mediates autophagic apoptosis in DCM by upregulating FoxO1 via ROS and enhancing FoxO1 acetylation through the suppression of SIRT1 activity. The discovery of this new mechanism provides new perspectives and potential therapeutic targets for understanding and treating DCM.

Electrochemical surface-enhanced Raman spectroscopy (E-SERS) of novel biodegradable ionic liquids.

Electrochemical SERS (E-SERS) was used for the first time to study the interfacial behavior of a class of pyridinium-based biodegradable ionic liquids at a silver nanoparticle (AgNP) electrode surface. An isomeric series of ionic liquids (IL) based on 3-butoxycarbonyl-1-methylpyridinium bis(trifluoromethanesulfonyl)imide were prepared, which have demonstrable biodegradability. It was found that all four of the isomeric ionic liquids studied exhibited excellent electrochemical stability as binary mixtures combined with methanol, with the absence of any specific redox processes occurring over nearly 3.0 V of applied potential. Normal Raman measurements of the neat isobutyl IL showed a signal rich in vibrational features, with strong contributions from both the anion and the bulky organic cation. E-SERS of the neat isobutyl IL was shown to exhibit excellent potential stability, with no potential-induced orientational change at the metal surface. When the ionic liquids were prepared as methanolic binary mixtures, dissociation of the IL ions was observed, and only the organic cation was shown to adsorb at the Ag/solution interface. The nature of the substituent on the ester group of the IL series was observed to have a significant effect on the orientation of the cation on the metal surface, based on the application of the metal surface selection rules combined with computational data. Notably, the isobutyl and sec-butyl isomers were observed to have an orientation wherein the pyridinium ring was oriented perpendicular to the surface, while the tert-butyl and n-butyl isomers were observed to have an orientation wherein the pyridinium ring was lying flat on the metal surface.

Prognostic Impact of Serum CRP Level in Head and Neck Squamous Cell Carcinoma.

Objective: This study evaluated the association of pretreatment serum C-reactive protein (CRP) level with prognosis in patients with head and neck squamous cell carcinoma (HNSCC). Methods: Within a single-center retrospective study, HNSCC patients receiving treatment between 2014 and 2016 were analyzed regarding the prognostic value of CRP serum levels. X-Tile software was used to determine the optimal cutoff value of serum CRP level. The log-rank test and Kaplan-Meier method were used to assess the effects of CRP level on prognosis in patients with HNSCC. Univariate and multivariate analyses (enter method) using a Cox proportional hazards model were utilized to identify prognostic indicators of progression-free survival (PFS) as the primary outcome and overall survival (OS) as the secondary outcome. Results: A total of 221 patients with HNSCC were assessed for eligibility, and 208 cases were included in the analysis. The HNSCC patients in the low-group (CRP ≤11.3 mg/L) showed better survival than those in the high-group (CRP > 11.3 mg/L). The univariate and multivariate analyses showed that N1-3 stage and a high serum CRP level (>11.3 mg/L) were unfavorable prognostic factors for PFS and OS in patients with HNSCC. Conclusion: Serum CRP level is an independent prognostic marker for patients with HNSCC. CRP level could be regarded as a novel prognostic factor for HNSCC patients.

Head and Neck Squamous Cell Carcinoma: NT5E Could Be a Prognostic Biomarker.

Head and neck squamous cell carcinoma (HNSCC) is a type of tumour with a relatively poor prognosis. In recent years, immune checkpoint inhibitors, such as CTLA-4 and PD-1/PDL-1 inhibitors, have improved the treatment status of advanced tumours. However, the emergence of drug resistance has brought difficulties to clinical treatment, and new immune checkpoint research is imminent. The hypoxia-adenosine pathway, in which CD73 encoded by the NT5E gene is a key enzyme for adenosine production, has been identified as an immune checkpoint of great potential. Therefore, NT5E may play an important role in HNSCC. We performed a detailed bioinformatics analysis of NT5E in HNSCC, and the results showed that the overexpression of NT5E in HNSCC was associated with poor prognosis. Our further investigation of the coexpression pattern of HNSCC could provide a reference for drug resistance and immunotherapy studies.

Sestrin2 is an endogenous antioxidant that improves contractile function in the heart during exposure to ischemia and reperfusion stress.

Sestrin2 (Sesn2) is a stress-inducible protein that plays a critical role in the response to ischemic stress. We recently recognized that Sesn2 may protect the heart against ischemic insults by reducing the generation of reactive oxygen species (ROS). After 45 min of ischemia followed by 24 h of reperfusion, myocardial infarcts were significantly larger in Sesn2 KO hearts than in wild-type hearts. Isolated cardiomyocytes from wild-type hearts treated with hypoxia and reoxygenation (H/R) stress showed significantly greater Sesn2 levels, compared with normoxic hearts (p < 0.05). Intriguingly, the administration of adeno-associated virus 9-Sesn2 into Sesn2 knockout (KO) hearts rescued Sesn2 protein levels and significantly improved the cardiac function of Sesn2 KO mice exposed to ischemia and reperfusion. The rescued levels of Sesn2 in Sesn2 KO hearts significantly ameliorated ROS generation and the activation of ROS-related stress signaling pathways during ischemia and reperfusion. Moreover, the rescued Sesn2 levels in Sesn2 KO cardiomyocytes improved the maximal velocity of cardiomyocyte shortening by H/R stress. Rescued Sesn2 levels also improved peak height, peak shortening amplitude, and maximal velocity of the re-lengthening of Sesn2 KO cardiomyocytes subjected to H/R. Finally, the rescued Sesn2 levels significantly augmented intracellular calcium levels and reduced the mean time constant of transient calcium decay in Sesn2 KO cardiomyocytes exposed to H/R. Overall, these findings indicated that Sesn2 can act as an endogenous antioxidant to maintain intracellular redox homeostasis under ischemic stress conditions.

The Ganglioside Monosialotetrahexosylganglioside Protects Auditory Hair Cells Against Neomycin-Induced Cytotoxicity Through Mitochondrial Antioxidation: An in vitro Study.

Neomycin is a common ototoxic aminoglycoside antibiotic that causes sensory hearing disorders worldwide, and monosialotetrahexosylganglioside (GM1) is reported to have antioxidant effects that protect various cells. However, little is known about the effect of GM1 on neomycin-induced hair cell (HC) ototoxic damage and related mechanism. In this study, cochlear HC-like HEI-OC-1 cells along with whole-organ explant cultures were used to establish an in vitro neomycin-induced HC damage model, and then the apoptosis rate, the balance of oxidative and antioxidant gene expression, reactive oxygen species (ROS) levels and mitochondrial membrane potential (MMP) were measured. GM1 could maintain the balance of oxidative and antioxidant gene expression, inhibit the accumulation of ROS and proapoptotic gene expression, promoted antioxidant gene expression, and reduce apoptosis after neomycin exposure in HEI-OC-1 cells and cultured cochlear HCs. These results suggested that GM1 could reduce ROS aggregation, maintain mitochondrial function, and improve HC viability in the presence of neomycin, possibly through mitochondrial antioxidation. Hence, GM1 may have potential clinical value in protecting against aminoglycoside-induced HC injury.

Expression Patterns of Three Important Hormone Genes and Respiratory Metabolism in Antheraea pernyi during Pupal Diapause under a Long Photoperiod.

The Chinese oak silkworm is commonly used in pupal diapause research. In this study, a long photoperiod was used to trigger pupal diapause termination. Genes encoding three hormones, namely prothoracicotropic hormone (PTTH), ecdysis triggering hormone (ETH), and eclosion hormone (EH), were studied. Additionally, ecdysteroids (mainly 20-hydroxyecdysone, 20E) were quantified by HPLC. Pupal diapause stage was determined by measuring respiratory intensity. The pupae enter a low metabolic rate, which starts approximately 1 month after pupal emergence. ApPTTH expression showed a small increase at 14 days and then a larger increase from 35 days under the long photoperiod treatment. A similar pattern was observed for the titer of 20E in the hemolymph. However, ApETH expression later increased under the long photoperiod treatment (42 days) just before eclosion. Moreover, ApEH expression increased from 21 to 35 days, and then decreased before ecdysis. These results suggest that hormone-related gene expression is closely related to pupal development. Our study lays a foundation for future diapause studies in A. pernyi.

The Multifaceted Role of Astrocyte Connexin 43 in Ischemic Stroke Through Forming Hemichannels and Gap Junctions.

Ischemic stroke is a multi-factorial cerebrovascular disease with high worldwide morbidity and mortality. In the past few years, multiple studies have revealed the underlying mechanism of ischemia/reperfusion injury, including calcium overload, amino acid toxicity, oxidative stress, and inflammation. Connexin 43 (Cx43), the predominant connexin protein in astrocytes, has been recently proven to display non-substitutable roles in the pathology of ischemic stroke development and progression through forming gap junctions and hemichannels. Under normal conditions, astrocytic Cx43 could be found in hemichannels or in the coupling with other hemichannels on astrocytes, neurons, or oligodendrocytes to form the neuro-glial syncytium, which is involved in metabolites exchange between communicated cells, thus maintaining the homeostasis of the CNS environment. In ischemic stroke, the phosphorylation of Cx43 might cause the degradation of gap junctions and the opening of hemichannels, contributing to the release of inflammatory mediators. However, the remaining gap junctions could facilitate the exchange of protective and harmful metabolites between healthy and injured cells, protecting the injured cells to some extent or damaging the healthy cells depending on the balance of the exchange of protective and harmful metabolites. In this study, we review the changes in astrocytic Cx43 expression and distribution as well as the influence of these changes on the function of astrocytes and other cells in the CNS, providing new insight into the pathology of ischemic stroke injury; we also discuss the potential of astrocytic Cx43 as a target for the treatment of ischemic stroke.

Determination of Alternaria toxins in drinking water by ultra-performance liquid chromatography tandem mass spectrometry.

A sensitive and reliable analytical method has been developed and validated for the determination of five Alternaria toxins, including tenuazonic acid (TeA), alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), and tentoxin (TEN), in drinking water using a one-step enrichment and clean-up strategy followed by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Drinking water samples were preprocessed using excess sodium sulfite to remove residual chlorine, and the pH was adjusted by formic acid. Analytes were concentrated and purified from the water samples using hydrophilic-lipophilic balanced (HLB) solid-phase extraction (SPE) cartridges. Chromatographic separation was performed on an Acquity HSS C18 column using 0.1 mM ammonium carbonate and methanol as the mobile phase. The average recoveries at three spiked levels (0.1, 0.5, and 1 ng/L for TeA, AOH, and ALT; 0.01, 0.05, and 0.1 ng/L for TEN and AME) were 76.1-106.5%, with an intra-day precision less than 15.5% and inter-day precision of 11.8-16.5%. The limits of detection (LODs) were 0.05 ng/L for TeA, AOH, and ALT and 0.005 ng/L for TEN and AME. The limits of quantification (LOQs) were 0.1 ng/L for TeA, AOH, and ALT and 0.01 ng/L for TEN and AME. The developed method was applied to monitor 289 drinking water samples collected in Beijing from 2015 to 2017, and TeA and TEN were found in 28 samples, with concentrations ranging from 0.16 to 2.7 ng/L and 0.21 to 2.2 ng/L, respectively.

ß-catenin regulates IRF3-mediated innate immune signalling in colorectal cancer.

OBJECTIVE: ß-catenin is one of the most critical oncogenes associated with many kinds of human cancers, especially in the human CRC. Innate immunity recognizes tumour derived damage-associated molecular patterns (DAMPs) and primes the anti-tumour adaptive responses. While the function of ß-catenin in CRC tumourigenesis is well established, its impact on innate immune evasion is largely unknown. The aim of this study is to characterize the role of ß-catenin in inhibiting RIG-I-like receptor (RLR)-mediated IFN-ß signalling in colorectal cancer. MATERIALS AND METHODS: Immunohistochemical staining and western blotting were conducted to study the expression of ß-catenin, IRF3 and phospho-IRF3 (p-IRF3) in CRC samples and cell lines. Plaque assay determining virus replication was performed to assess the regulation of ß-catenin on IFN-ß signalling. The inhibition of ß-catenin on RLR-mediated IFN-ß signalling was further studied by real-time analyses and reporter assays in the context of lentiviral-mediated ß-catenin stably knocking down. Lastly, co-immunoprecipitation and nuclear fractionation assay were conducted to monitor the interaction between ß-catenin and IRF3. RESULTS: We found that high expression of ß-catenin positively correlated with the expression of IRF3 in CRC cells. Overexpression of ß-catenin increased the viral replication. Conversely knocking down of ß-catenin inhibited viral replication. Furthermore, our data demonstrated that ß-catenin could inhibit the expression of IFN-ß and interferon-stimulated gene 56 (ISG56). Mechanistically, we found that ß-catenin interacted with IRF3 and blocked its nuclear translocation. CONCLUSION: Our study reveals an unprecedented role of ß-catenin in enabling innate immune evasion in CRC.

Biomarkers of cigarette smoking and DNA methylating agents: Raman, SERS and DFT study of 3-methyladenine and 7-methyladenine.

3-Methyladenine and 7-methyladenine are biomarkers of DNA damage from exposure to methylating agents. For example, the concentration of 3-methyladenine increases significantly in the urine of cigarette smokers. Surface-enhanced Raman spectroscopy (SERS) has shown much potential for detection of biomolecules, including DNA. Much work has been dedicated to the canonical nucleobases, with comparatively fewer investigations of modified DNA and modified DNA nucleobases. Herein, Raman spectroscopy and SERS are used to examine the adsorption orientations of 3-methyladenine and 7-methyladenine on Ag nanoparticles. Density functional theory (DFT) calculations at the B3LYP level are used to support the conclusions via simulated spectra of the nucleobases and of Ag+/nucleobase complexes. The results herein show that 7-methyladenine adsorbs upright via its N3 and N9 atoms side, similarly to adenine. 3-Methyladenine adsorbs in a very tilted or flat orientation on the Ag nanoparticles. These findings will be useful for future SERS or other nanoparticle-based bioanalytical assays for detection of these methyladenines or other modified nucleobases.

Hyper activation of ß-catenin signalling induced by IKKε inhibition thwarts colorectal cancer cell proliferation.

OBJECTIVE: Aberrant activation of Wnt/ß-catenin signalling contributes significantly to the development of human colorectal cancers and ß-catenin is the key signalling molecule transducing canonical Wnt/ß-catenin signalling. Therefore, ß-catenin is a promising therapeutic target for cancer treatment. This study demonstrates that the oncogenic IKKε kinase phosphorylates ß-catenin to restrain its hyper activation, therefore promoting colorectal cancer (CRC) cell proliferation. MATERIALS AND METHODS: IKKε and ß-catenin expression levels in human colorectal cancer tissues and cell lines were analysed by immunohistochemical staining and Western blotting. The regulation of IKKε on Wnt/ß-catenin signalling pathway was studied by reporter assay and real-time PCR analysis in the context of IKKε stably knocking down. Co-immunoprecipitation was conducted to monitor the interaction between IKKε and ß-catenin. Kinase assay was performed to measure ß-catenin post-translational modifications induced by IKKε. RESULTS: Oncogenic IKKε kinase is required for the proliferation of colorectal cancer cells. Mechanistically, inhibition of IKKε results in ß-catenin hyper activation and thwarts CRC cell proliferation. Furthermore, IKKε phosphorylates ß-catenin and inhibits the activation of ß-catenin signalling. CONCLUSION: Our study suggests that IKKε is a potential target to combat CRC induced by aberrant Wnt/ß-catenin signalling.