To explore the relationship between plasma microRNA126 (miR-126) level and coronary collateral circulation (CCC) formation and to determine whether the miR-126 in plasma could serve as a blood-based biomarker for CCC in patients with severely narrowed coronary arteries (CAD).
Ataxia-telangiectasia mutated (ATM), a member of the phosphatidylinositol 3 kinase-like kinase family, is a master regulator of the double strand DNA break-repair pathway after genotoxic stress. Here, we found ATM serves as an essential regulator of TNF-induced NF-kB pathway. We observed that TNF exposure of cells rapidly induced DNA double strand breaks and activates ATM. TNF-induced ROS promote nuclear IKK? association with ubiquitin and its complex formation with ATM for nuclear export. Activated cytoplasmic ATM is involved in the selective recruitment of the E3-ubiquitin ligase ?-TrCP to phospho-I?B? proteosomal degradation. Importantly, ATM binds and activates the catalytic subunit of protein kinase A (PKAc), ribosmal S6 kinase that controls RelA Ser 276 phosphorylation. In ATM knockdown cells, TNF-induced RelA Ser 276 phosphorylation is significantly decreased. We further observed decreased binding and recruitment of the transcriptional elongation complex containing cyclin dependent kinase-9 (CDK9; a kinase necessary for triggering transcriptional elongation) to promoters of NF-?B-dependent immediate-early cytokine genes, in ATM knockdown cells. We conclude that ATM is a nuclear damage-response signal modulator of TNF-induced NF-?B activation that plays a key scaffolding role in I?B? degradation and RelA Ser 276 phosphorylation. Our study provides a mechanistic explanation of decreased innate immune response associated with A-T mutation.
High-normal blood pressure is considered a precursor of stage 1 hypertension that is associated with metabolic disorders. This study aims to investigate whether the pharmacologic treatment of high-normal blood pressure affects metabolism, especially in abdominally obese individuals, and the pharmacoeconomics of two antihypertensive agents, telmisartan and indapamide. Subjects with high-normal blood pressure were randomly assigned to receive telmisartan, indapamide or placebo for 3 years. All the subjects were instructed to modify their lifestyle to reduce blood pressure throughout the study. A total of 221 subjects were randomly assigned to telmisartan, 213 to indapamide and 230 to placebo. After the 3-year intervention, blood pressure was lower in the telmisartan and indapamide groups (P<0.05), FPG in the telmisartan group was lower during the first 2 years (P<0.05) and no characteristic differences were found in those with abdominal obesity among the three groups (P>0.05). The percentage of subjects with metabolic syndrome was significantly decreased in the telmisartan and indapamide groups (P<0.05), but was only significantly decreased in the telmisartan group for subjects with abdominal obesity (P<0.05). The acquisition cost for telmisartan was ~1.86 times higher than for indapamide for a similar antihypertensive effect. The intervention for high-normal blood pressure with telmisartan and indapamide appeared to be feasible and reduced the risk of metabolic syndrome. Telmisartan was more effective, whereas indapamide had better pharmacoeconomic benefits.Hypertension Research advance online publication, 2 October 2014; doi:10.1038/hr.2014.148.
We evaluated the impact of clopidogrel 150 mg/d in patients with chronic kidney disease (CKD) having clopidogrel resistance (CR) after percutaneous coronary intervention (PCI); 1076 consecutive patients with coronary artery disease (CAD) having CKD were enrolled. Maximal platelet aggregation (MPA) was assessed before, 24 hours, and 30 days after a 300-mg loading dose of clopidogrel prior to PCI. After PCI, 370 patients with CR were randomized to receive clopidogrel 75 mg/d (n = 184) or 150 mg/d (n = 186) for 30 days. Stent thrombosis (ST), major adverse cardiac events (MACEs), and bleeding were analyzed after 1 month. Patients in the 150 mg group had significant lower rates of ST and MACE. There was no significant difference in major or minor bleeding. Patients in the 150 mg group had lower MPA and greater inhibition of platelet aggregation. One-month administration of 150 mg/d of clopidogrel decreases the rate of ST and MACE without increasing bleeding in patients with CKD having CR after PCI.
Asthma is an idiopathic disease associated with episodic inflammation and reversible airway obstruction that is triggered by environmental agents. Allergic and infectious agents trigger asthmatic exacerbations through the innate immune response (IIR). The IIR is activated by sentinel cells in the airways to elaborate inflammatory cytokines and protective mucosal interferons whose actions are designed to limit the spread of the organism, as well as to activate the adaptive immune response. We address the structure of the IIR pathway in sentinel cells of the airway and describe observations on its dysregulation. The IIR is triggered in a cell-type specific manner by germline-encoded pathogen recognition receptors (PPRs) including plasma membrane Toll-like receptors (TLRs) and the cytoplasmic Retinoic Acid-inducible Gene (RIG)-I-like RNA helicases, and protein kinase R (PKR). Although their mechanisms of intracellular signaling differ, both pathways converge on a small group of transcriptional effectors, nuclear factor-?B (NF-?B), IFN regulatory factor (IRF), and signal transducer and activator of transcription (STAT). We describe several distinct techniques to quantitate the IIR including assays based on quantitative real-time PCR (Q-RT-PCR) of NF-?B and IRF3-regulated genes, multiplex bead-based analysis of secreted proteins/cytokines and more recent developments in targeted, quantitative selected reaction monitoring (SRM)-mass spectrometry (MS). Application of these methods for quantitation of the IIR will further our understanding of the role of the IIR in asthma and its contribution to disease heterogeneity.
Abstract Background: Visit-to-visit variability in blood pressure (BP) was demonstrated to correlate with cardiovascular events independent of mean BP. The goal of the present study was to investigate the correlation of visit-to-visit BP variability with artery stiffness and myocardial perfusion in on-treated hypertensive patients. Methods: BP was measured in 271 hypertensive patients at every visit over the course of the antihypertensive treatment, and the standard deviation (SD), coefficient of variation (CV), maximum, and minimum in serial BP were calculated. Non-invasive pulse wave analysis was performed in all patients. Results: Compared with baseline, carotid-femoral pulse wave velocity (cfPWV), aortic augmentation index (Aix) and Aix adjusted to a "standard heart rate" of 75 beats/min (Aix@HR75) were markedly declined, and sub-endocardial viability ratio (SEVR) was obviously increased in each group (p?0.001). The changes of cfPWV, SEVR, Aix and Aix@HR75 in patients with lower SD of systolic blood pressure (SBP) were significantly greater than those in patients with higher SD of SBP. And the changes were statistically correlated with both SD and CV of serial SBP during follow-up, even after adjusted for mean SBP and mean diastolic blood pressure (DBP). Conclusion: Visit-to-visit SBP variability is independently correlated with changes of artery stiffness and myocardial perfusion in on-treated hypertensive patients.
Signal Transducers and Activator of Transcription-3 (STAT3) are latent transcription factors that are regulated by post-translational modifications (PTMs) in response to cellular activation by the IL-6 superfamily of cytokines to regulate cell cycle progression and/or apoptosis. Here we observe that STAT3 is inducibly mono-ubiquitinated and investigate its consequences. Using domain mapping and highly specific selected reaction monitoring-mass spectrometric assays, we identify lysine (K) 97 in its NH2-terminal domain as the major mono-ubiquitin conjugation site. We constructed a mono-ubiquitinated mimic consisting of a deubiquitinase-resistant monomeric ubiquitin fused to the NH2 terminus of STAT3 (ubiquitinated-STAT3 FP). In complex assays of ectopically expressed ubi-STAT3-FP, we observed enhanced complex formation with bromodomain-containing protein 4 (BRD4), a component of the activated positive transcriptional elongation factor (P-TEFb) complex. Chromatin immunoprecipitation experiments in STAT3(+/-) and STAT3(-/-) MEFs showed BRD4 recruitment to STAT3-dependent suppressor of cytokine signaling-3 gene (SOCS3). The effect of a selective small molecule inhibitor of BRD4, JQ1, to inhibit SOCS3 expression demonstrated the functional role of BRD4 for STAT3-dependent transcription. Additionally, ectopic ubiquitinated-STAT3 FP expression upregulated BCL2, BCL2L1, APEX1, SOD2, CCND1 and MYC expression indicating the role of ubiquitinated STAT3 in anti-apoptosis and cellular proliferation. Finally we observed that ubiquitinated-STAT3 FP suppressed TNF?-induced apoptotic cell death, indicating the functional importance of mono-ubiquitinated STAT3 in antiapoptotic gene expression. We conclude that STAT3 mono-ubiquitination is a key trigger in BRD4-dependent antiapoptotic and pro-proliferative gene expression programs. Thus, inhibiting the STAT3 mono-ubiquitination-BRD4 pathway may be a novel therapeutic target for the treatment of STAT3-dependent proliferative diseases.
Modulators (Ms) are proteins that modify the activity of transcription factors (TFs) and influence expression of their target genes (TGs). To discover modulators of NF-?B/RelA, we first identified 365 NF-?B/RelA-binding proteins using liquid chromatography-tandem mass spectrometry (LC-MS/MS). We used a probabilistic model to infer 8349 (M, NF-?B/RelA, TG) triplets and their modes of modulatory action from our combined LC-MS/MS and ChIP-Seq (ChIP followed by next generation sequencing) data, published RelA modulators and TGs, and a compendium of gene expression profiles. Hierarchical clustering of the derived modulatory network revealed functional subnetworks and suggested new pathways modulating RelA transcriptional activity. The modulators with the highest number of TGs and most non-random distribution of action modes (measured by Shannon entropy) are consistent with published reports. Our results provide a repertoire of testable hypotheses for experimental validation. One of the NF-?B/RelA modulators we identified is STAT1. The inferred (STAT1, NF-?B/RelA, TG) triplets were validated by LC-selected reaction monitoring-MS and the results of STAT1 deletion in human fibrosarcoma cells. Overall, we have identified 562 NF-?B/RelA modulators, which are potential drug targets, and clarified mechanisms of achieving NF-?B/RelA multiple functions through modulators. Our approach can be readily applied to other TFs.
Trimetazidine has been shown to improve angina pectoris and left ventricular (LV) function in diabetic patients with ischaemic cardiomyopathy. The objective of this study was to evaluate the effects of trimetazidine on recurrent angina pectoris and LV structure after drug-eluting stent (DES) implantation in elderly multivessel coronary heart disease (CHD) patients with diabetes mellitus (DM) and a left ventricular ejection fraction (LVEF) of ? 50 %.
Hippocampal network hyperexcitability is considered an early indicator of Alzheimer's disease (AD) memory impairment. Some AD mouse models exhibit similar network phenotypes. In this study we focused on dentate gyrus (DG) granule cell spontaneous and evoked properties in 9-month-old Tg2576 mice that model AD amyloidosis and cognitive deficits. Using whole-cell patch-clamp recordings, we found that Tg2576 DG granule cells exhibited spontaneous EPSCs that were higher in frequency but not amplitude compared with wild-type mice, suggesting hyperactivity of DG granule cells via a presynaptic mechanism. Further support of a presynaptic mechanism was revealed by increased I-O relationships and probability of release in Tg2576 DG granule cells. Since we and others have shown that activation of the peroxisome proliferator-activated receptor gamma (PPAR?) axis improves hippocampal cognition in mouse models for AD as well as benefitting memory performance in some humans with early AD, we investigated how PPAR? agonism affected synaptic activity in Tg2576 DG. We found that PPAR? agonism normalized the I-O relationship of evoked EPSCs, frequency of spontaneous EPSCs, and probability of release that, in turn, correlated with selective expression of DG proteins essential for presynaptic SNARE function that are altered in patients with AD. These findings provide evidence that DG principal cells may contribute to early AD hippocampal network hyperexcitability via a presynaptic mechanism, and that hippocampal cognitive enhancement via PPAR? activation occurs through regulation of presynaptic vesicular proteins critical for proper glutamatergic neurotransmitter release, synaptic transmission, and short-term plasticity.
We present an integrated dynamical cross-talk model of the epithelial innate immune response (IIR) incorporating RIG-I and TLR3 as the two major pattern recognition receptors (PRR) converging on the RelA and IRF3 transcriptional effectors. bioPN simulations reproduce biologically relevant gene-and protein abundance measurements in response to time course, gene silencing and dose-response perturbations both at the population and single cell level. Our computational predictions suggest that RelA and IRF3 are under auto- and cross-regulation. We predict, and confirm experimentally, that RIG-I mRNA expression is controlled by IRF7. We also predict the existence of a TLR3-dependent, IRF3-independent transcription factor (or factors) that control(s) expression of MAVS, IRF3 and members of the IKK family. Our model confirms the observed dsRNA dose-dependence of oscillatory patterns in single cells, with periods of 1-3 hr. Model fitting to time series, matched by knockdown data suggests that the NF-?B module operates in a different regime (with different coefficient values) than in the TNF?-stimulation experiments. In future studies, this model will serve as a foundation for identification of virus-encoded IIR antagonists and examination of stochastic effects of viral replication. Our model generates simulated time series, which reproduce the noisy oscillatory patterns of activity (with 1-3 hour period) observed in individual cells. Our work supports the hypothesis that the IIR is a phenomenon that emerged by evolution despite highly variable responses at an individual cell level.
To investigate variability in self-measured home blood pressure (HBP) and its effects on carotid artery atherosclerosis and endothelial function in normotensive and mild-moderate hypertensive individuals.
There is conflicting evidence regarding the effectiveness of remote ischemic preconditioning (RIPC) in patients undergoing elective percutaneous coronary intervention (PCI). Therefore, we prospectively enrolled elderly patients with coronary heart disease (CHD) with diabetes mellitus (DM) undergoing elective drug-eluting stent (DES) implantation. They were randomized to receive RIPC within 2 hours before PCI (n = 102) or not (controls, n = 98). Baseline clinical characteristics were similar between the 2 groups. Despite a trend toward decline, the median high-sensitivity cardiac troponin I (hscTnI) level (P = .256) and the incidence of myocardial infarction (MI) type 4a (P = .106) in the RIPC group 16 hours after PCI procedure was not significantly different from the control group. The RIPC could attenuate the release of a myocardial biomarker but failed to show a significant effect on hscTnI level or MI type 4a incidence after PCI procedure in elderly patients with CHD having DM undergoing elective DES implantation.
The effect of statins in patients with acute coronary syndrome (ACS) at advanced age with lower low-density lipoprotein cholesterol (LDL-C) levels undergoing percutaneous coronary intervention (PCI) remains unknown. We evaluated the effect of statins in 220 Chinese patients with ACS aged ?75 years with low LDL-C undergoing PCI. Biomarkers were measured before and 6 hours after PCI, and patients were followed up for 1 year. Biomarkers in the statin group at 6 hours post-PCI were lower than controls (creatine kinase-myocardial band 14.2 ± 5.78 vs 47.3 ± 16.4 IU/L, P = .03; cardiac troponin I 0.36 ± 0.12 vs 1.33 ± 0.47 ng/mL, P = .01; and high-sensitivity C-reactive protein 7.6 ± 4.3 vs 13.6 ± 4.5 mg/L, P = .001, respectively). Significant differences were found in major adverse cardiac events at 1 year (P = .02-.01), while target lesion revascularization alone was less at 3 months between the 2 groups (P = .03). This study demonstrates that elderly patients with ACS having low LDL-C benefit from statins regardless of type, dosage, and duration of statin administration prior to PCI.
A heterotrophic/biofilm-electrode autotrophic denitrification reactor (HAD-BER) was developed to improve denitrification efficiency and reduce the consumption of organic carbon source. Maximum nitrate removal efficiency of 99.9% was gained under the optimum current density of 200 mA/m(2). The number of heterotrophic denitrification bacteria (HDB) 2.0 × 10(5) and hydrogen autotrophic denitrification bacteria (ADB) 2.0 × 10(3) in per milliliter biofilm solution were observed by the most probable number (MPN) culture, demonstrating that HDB and ADB coexist in the HAD-BER. The inorganic carbon source for autotrophic denitrification was supplied by the dissolved carbon dioxide (CO2) evolved from the heterotrophic denitrification process, indicating that there was synergistic interaction between the HDB and ADB, i.e., the organic carbon source used for denitrification could be decreased in the HAD-BER. Therefore, the developed HAD-BER would be a promising approach for nitrate removal from groundwater.
Respiratory syncytial virus (RSV) is a negative-sense single-stranded RNA virus responsible for lower respiratory tract infections. During infection, the presence of double-stranded RNA (dsRNA) activates the interferon (IFN) regulatory factor 3 (IRF3) transcription factor, an event triggering expression of immediate early, IFN-stimulated genes (ISGs). We examine the role of transcriptional elongation in control of IRF3-dependent ISG expression. RSV infection induces ISG54, ISG56, and CIG5 gene expression in an IRF3-dependent manner demonstrated by IRF3 small interfering RNA (siRNA) silencing in both A549 epithelial cells and IRF3(-/-) MEFs. ISG expression was mediated by the recruitment of IRF3, CDK9, polymerase II (Pol II), and phospho-Ser(2) carboxy-terminal domain (CTD) Pol II to the IFN-stimulated response element (ISRE) binding sites of the IRF3-dependent ISG promoters in native chromatin. We find that RSV infection enhances the activated fraction of cyclin-dependent kinase 9 (CDK9) by promoting its association with bromodomain 4 (BRD4) and disrupting its association with the inhibitory 7SK small nuclear RNA. The requirement of CDK9 activity for ISG expression was shown by siRNA-mediated silencing of CDK9 and by a selective CDK9 inhibitor in A549 cells. In contrast, RSV-induced beta interferon (IFN-?) expression is not influenced by CDK9 inhibition. Using transcript-selective quantitative real-time reverse transcription-PCR (Q-RT-PCR) assays for the ISG54 gene, we observed that RSV induces transition from short to fully spliced mRNA transcripts and that this transition is blocked by CDK9 inhibition in both A549 and primary human small airway epithelial cells. These data indicate that transcription elongation plays a major role in RSV-induced ISG expression and is mediated by IRF3-dependent recruitment of activated CDK9. CDK9 activity may be a target for immunomodulation in RSV-induced lung disease.
The NF-?B transcription factor mediates the inflammatory response through distinct (canonical and non-canonical) signaling pathways. The mechanisms controlling utilization of either of these pathways are largely unknown. Here we observe that TNF stimulation induces delayed NF-?B2/p100 processing and investigate the coupling mechanism. TNF stimulation induces TNF-associated factor-1 (TRAF-1) that directly binds NF-?B-inducing kinase (NIK) and stabilizes it from degradation by disrupting its interaction with TRAF2·cIAP2 ubiquitin ligase complex. We show that TRAF1 depletion prevents TNF-induced NIK stabilization and reduces p52 production. To further examine the interactions of TRAF1 and NIK with NF-?B2/p100 processing, we mathematically modeled TRAF1·NIK as a coupling signaling complex and validated computational inference by siRNA knockdown to show non-canonical pathway activation is dependent not only on TRAF1 induction but also NIK stabilization by forming TRAF1·NIK complex. Thus, these integrated computational-experimental studies of TNF-induced TRAF1 expression identified TRAF1·NIK as a central complex linking canonical and non-canonical pathways by disrupting the TRAF2-cIAP2 ubiquitin ligase complex. This feed-forward kinase pathway is essential for the activation of non-canonical pathway.
We evaluated the impact of transradial coronary procedures on the vasodilatory function of the radial artery. A total of 65 patients who underwent transradial coronary procedures were enrolled. All patients were examined with B-mode high-resolution ultrasound. Radial artery baseline diameter and response to flow-mediated dilation (FMD) and nitroglycerin-mediated dilation (NMD) were measured in the right radial artery. The FMD of the right radial artery was 11.5%, 4.1%, and 0.7%, respectively, before the procedures, 1 day, and 3 months after the procedures (P < .05 at 1 day, P < .01 at 3 months). The NMD of the right radial artery was 17.6%, 5.4%, and 6.3%, respectively, before the procedures, 1 day, and 3 months after the procedures (P < .05 at 1 day, P < .05 at 3 months). Transradial coronary procedures decrease radial artery FMD and NMD resulting in immediate and persistent blunting of vasodilatory function.
The authors investigated effects of excessive salt intake and potassium supplementation on ambulatory arterial stiffness index (AASI) and endothelin-1 (ET-1) in salt-sensitive and non-salt-sensitive individuals. AASI and symmetric AASI (s-AASI) were used as indicators of arterial stiffness. Plasma ET-1 levels were used as an index of endothelial function. Chronic salt-loading and potassium supplementation were studied in 155 normotensive to mild hypertensive patients from rural northern China. After 3 days of baseline investigation, participants were maintained sequentially for 7 days each on diets of low salt (51.3 mmol/d), high salt (307.7 mmol/d), and high salt+potassium (60 mmol/d). Ambulatory 24-hour blood pressure (BP) and plasma ET-1 were measured at baseline and on the last 2 days of each intervention. High-salt intervention significantly increased BP, AASI, s-AASI (all P<.001); potassium supplementation reversed increased plasma ET-1 levels. High-salt-induced changes in BP, s-AASI, and plasma ET-1 were greater in salt-sensitive individuals. Potassium supplementation decreased systolic BP and ET-1 to a significantly greater extent in salt-sensitive vs non-salt-sensitive individuals (P<.001). Significant correlations were identified between s-AASI and ET-1 change ratios in response to both high-salt intervention and potassium supplementation (P<.001). Reducing dietary salt and increasing daily potassium improves arterial compliance and ameliorates endothelial dysfunction.
The innate immune response (IIR) is a coordinated intracellular signaling network activated by the presence of pathogen-associated molecular patterns that limits pathogen spread and induces adaptive immunity. Although the precise temporal activation of the various arms of the IIR is a critical factor in the outcome of a disease, currently there are no quantitative multiplex methods for its measurement. In this study, we investigate the temporal activation pattern of the IIR in response to intracellular double-stranded RNA stimulation using a quantitative 10-plex stable isotope dilution-selected reaction monitoring-MS assay. We were able to observe rapid activation of both NF-?B and IRF3 signaling arms, with IRF3 demonstrating a transient response, whereas NF-?B underwent a delayed secondary amplification phase. Our measurements of the NF-?B-I?B? negative feedback loop indicate that about 20% of I?B? in the unstimulated cell is located within the nucleus and represents a population that is rapidly degraded in response to double-stranded RNA. Later in the time course of stimulation, the nuclear I?B? pool is repopulated first prior to its cytoplasmic accumulation. Examination of the IRF3 pathway components shows that double-stranded RNA induces initial consumption of the RIG-I PRR and the IRF3 kinase (TBK1). Stable isotope dilution-selected reaction monitoring-MS measurements after siRNA-mediated IRF3 or RelA knockdown suggests that a low nuclear threshold of NF-?B is required for inducing target gene expression, and that there is cross-inhibition of the NF-?B and IRF3 signaling arms. Finally, we were able to measure delayed noncanonical NF-?B activation by quantifying the abundance of the processed (52 kDa) NF-?B2 subunit in the nucleus. We conclude that quantitative proteomics measurement of the individual signaling arms of the IIR in response to system perturbations is significantly enabled by stable isotope dilution-selected reaction monitoring-MS-based quantification, and that this technique will reveal novel insights into the dynamics and connectivity of the IIR.
A novel biosorbent derived from agricultural residue - walnut shell (WS) is reported to remove cesium from aqueous solution. Nickel hexacyanoferrate (NiHCF) was incorporated into this biosorbent, serving as a high selectivity trap agent for cesium. Field emission scanning electron microscope (FE-SEM) and thermogravimetric and differential thermal analysis (TG-DTA) were utilized for the evaluation of the developed biosorbent. Determination of kinetic parameters for adsorption was carried out using pseudo first-order, pseudo second-order kinetic models and intra-particle diffusion models. Adsorption equilibrium was examined using Langmuir, Freundlich and Dubinin-Radushkevich adsorption isotherms. A satisfactory correlation coefficient and relatively low chi-square analysis parameter ?(2) between the experimental and predicted values of the Freundlich isotherm demonstrate that cesium adsorption by NiHCF-WS is a multilayer chemical adsorption. Thermodynamic studies were conducted under different reaction temperatures and results indicate that cesium adsorption by NiHCF-WS is an endothermic (?H° > 0) and spontaneous (?G° < 0) process.
In the present study, a novel tablet porous material (TPM) was developed from Kanuma clay (K-clay), corn starch, and calcium oxide. Laboratory-scale batch experiments were conducted to evaluate the phosphate adsorption capacity of TPM from aqueous solution. The adsorption isotherms, adsorption kinetics, phosphate recycling, and major factors such as temperature, pH, and dosage were investigated. The phosphate adsorption results fitted the Freundlich isotherm model very well, and the adsorption process was an endothermic and spontaneous reaction which could be described by a pseudo second-order kinetic model. The maximum phosphate adsorption capacity was 4.39 mg g(-1), and its equilibrium could be attained in 2h. The solution pH had little effect on TPM phosphate removal when pH varied from 5.0 to 9.0. 70.29% of adsorbed phosphate could be recycled when 0.2N HCl was used as eluant, and the present developed TPM could be recovered and reused for 5 times. This novel developed TPM is a promising adsorbent than other clay mineral materials for phosphate removal from wastewater.
Quantitative measurement of the major regulatory proteins in signaling networks poses several technical challenges, including low abundance, the presence of post-translational modifications (PTMs), and the lack of suitable affinity detection reagents. Using the innate immune response (IIR) as a model signaling pathway, we illustrate the approach of stable isotope dilution (SID)-selected reaction monitoring (SRM)-mass spectrometry (MS) assays for quantification of low abundance signaling proteins. A work flow for SID-SRM-MS assay development is established for proteins with experimentally observed MS spectra and for those without. Using the interferon response factor (IRF)-3 transcription factor as an example, we illustrate the steps in high responding signature peptide identification, SID-SRM-MS assay optimization, and evaluation. SRM assays for normalization of IIR abundance to invariant housekeeping proteins are presented. We provide an example of SID-SRM assay development for post-translational modification (PTM) detection using an activating phospho-Ser modified NF-?B/RelA transcription factor, and describe challenges inherent in PTM-SID-SRM-MS assay development. Application of highly qualified quantitative, SID-SRM-MS assays will enable a systems-level approach to understanding the dynamics and kinetics of signaling in host cells, such as the IIR.
In this study, natural Akadama clay was used for Cr (VI) removal from aqueous solution. Batch experiments were carried out to investigate the effect of contact time, initial pH, and adsorbent dose on Cr (VI) adsorption. Results showed that Cr (VI) adsorption on natural Akadama clay reached equilibrium in 180 min. The Cr (VI) removal efficiency of 46.8% without pH adjustment increased to 73.8% at the optimum initial pH of 2. The maximum adsorption capacity was 4.29 mg g(-1) at an initial concentration of 50.0 mg L(-1) and adsorbent dosage of 5 g L(-1). The equilibrium data fitted Freundlich isotherm better than Langmuir isotherm, and they were well explained by pseudo-second-order kinetic model. Adsorption mechanism analysis proved that electrostatic adsorption dominated during the removal process. Results from this study demonstrate that natural Akadama clay has the potential to be an efficient adsorbent for Cr (VI) adsorption compared to other natural mineral adsorbents.
An intensified biofilm-electrode reactor (IBER) was developed to treat nitrate-contaminated drinking water. Different running conditions were conducted to investigate the behavior of autotrophic denitrification (AD) and heterotrophic denitrification (HD) in the IBER. In AD process, the nitrate nitrogen coulomb-reduction rate was used to evaluate the performance of the reactor. The maximum NO(3)(-)-N removal efficiency was 6.8% at the current of 60 mA, while nitrate nitrogen coulomb-reduction rate was 0.024 mg C(-1). The optimum conditions for HD process were C/N=0.8 and HRT=8h, under which complete NO(3)(-)-N removal and no NO(2)(-)-N accumulation were observed. With the cooperative effect of AD and HD in the heterotrophic and autotrophic denitrification (HAD) process, large treatment capacity, high denitrification efficiency, and low nitrite and ammonia accumulation were achieved. The results proved that HAD process was superior to single AD and HD for nitrate removal in the IBER.
Atherosclerosis is a chronic inflammatory disease regulated by T lymphocyte subsets. Th17 cells reportedly play important roles in the development of inflammatory and autoimmune diseases. In this study, we investigated the contributions of circulating Th17 cells and plasma Th17-associated cytokines to carotid artery plaques.
The transradial approach (TRA) is commonly applied for coronary catheterization. However, there are few reports on the safety and feasibility of transradial catheterization in patients with prior coronary artery bypass graft (CABG) surgery. We retrospectively evaluated 124 consecutive patients who underwent graft angiography and intervention via the transradial (TRA group, n = 68) or transfemoral approach (TFA group, n = 56). The baseline clinical characteristics between the 2 groups were similar except for prior myocardial infarction. No significant difference (P > .05)was observed in procedure time, the success rate of puncture, angiography, and intervention procedure between the 2 groups. There was no significant difference in major adverse cardiac and cerebrovascular events during hospitalization. However, the vascular access site complications were significantly lower (P = .021) and the duration of hospitalization was shorter (P = .007) in the TRA group. The TRA for coronary bypass graft angiography and intervention was safe and feasible.
Nuclear Factor-?B (NF-?B) is a family of inducible transcription factors regulated by stimulus-induced protein interactions. In the cytoplasm, the NF-?B member RelA transactivator is inactivated by binding inhibitory I?Bs, whereas in its activated state, the serine-phosphorylated protein binds the p300 histone acetyltransferase. Here we describe the isolation of a ssDNA aptamer (termed P028F4) that binds to the activated (I?B?-dissociated) form of RelA with a K(D) of 6.4 × 10(-10), and its application in an enrichment-mass spectrometric quantification assay. ssDNA P028F4 competes with cognate duplex high affinity NF-?B binding sites for RelA binding in vitro, binds activated RelA in eukaryotic nuclei and reduces TNF?-stimulated endogenous NF-?B dependent gene expression. Incorporation of P028F4 as an affinity isolation step enriches for serine 536 phosphorylated and p300 coactivator complexed RelA, simultaneously depleting I?B?·RelA complexes. A stable isotope dilution (SID)-selected reaction monitoring (SRM)- mass spectrometry (MS) assay for RelA was developed that produced a linear response over 1,000 fold dilution range of input protein and had a 200 amol lower limit of quantification. This multiplex SID-SRM-MS RelA assay was used to quantify activated endogenous RelA in cytokine-stimulated eukaryotic cells isolated by single-step P028F4 enrichment. The aptamer-SID-SRM-MS assay quantified the fraction of activated RelA in subcellular extracts, detecting the presence of a cytoplasmic RelA reservoir unresponsive to TNF? stimulation. We conclude that aptamer-SID-SRM-MS is a versatile tool for quantification of activated NF-?B/RelA and its associated complexes in response to pathway activation.
Metabolic syndrome (MetS) is regarded as a risk factor for coronary artery disease (CAD). However, the influence of MetS on morbidity and mortality after drug-eluting stent (DES) implantation in Chinese patients with CAD remains unknown. We evaluated the impact of MetS on the clinical outcome of 1224 patients following DES implantation. After a mean follow-up of 35.4 months, patients with MetS had a significantly higher incidence of all-cause death and major adverse cardiovascular events (MACE) compared with patients without MetS (P < .001). Analyses of individual MetS components showed that dysglycemia at the time of DES implantation predicted increased all-cause mortality, while the presence of hypertension and dysglycemia predicted increased incidence of MACE.
An intensified biofilm-electrode reactor (IBER) combining heterotrophic and autotrophic denitrification was developed for treatment of nitrate contaminated groundwater. The reactor was evaluated with synthetic groundwater (NO(3)(-)-N50 mg L(-1)) under different hydraulic retention times (HRTs), carbon to nitrogen ratios (C/N) and electric currents (I). The experimental results demonstrate that high nitrate and nitrite removal efficiency (100%) were achieved at C/N = 1, HRT = 8h, and I = 10 mA. C/N ratios were reduced from 1 to 0.5 and the applied electric current was changed from 10 to 100 mA, showing that the optimum running condition was C/N = 0.75 and I = 40 mA, under which over 97% of NO(3)(-)-N was removed and organic carbon (methanol) was completely consumed in treated water. Simultaneously, the denitrification mechanism in this system was analyzed through pH variation in effluent. The CO(2) produced from the anode acted as a good pH buffer, automatically controlling pH in the reaction zone. The intensified biofilm-electrode reactor developed in the study was effective for the treatment of groundwater polluted by nitrate.
We describe a method for ratio estimations in (18)O-water labeling experiments acquired from low resolution isotopically resolved data. The method is implemented in a software package specifically designed for use in experiments making use of zoom-scan mode data acquisition. Zoom-scan mode data allow commonly used ion trap mass spectrometers to attain isotopic resolution, which makes them amenable to use in labeling schemes such as (18)O-water labeling, but algorithms and software developed for high resolution instruments may not be appropriate for the lower resolution data acquired in zoom-scan mode. The use of power spectrum analysis is proposed as a general approach that may be uniquely suited to these data types. The software implementation uses a power spectrum to remove high-frequency noise and band-filter contributions from coeluting species of differing charge states. From the elemental composition of a peptide sequence, we generate theoretical isotope envelopes of heavy-light peptide pairs in five different ratios; these theoretical envelopes are correlated with the filtered experimental zoom scans. To automate peptide quantification in high-throughput experiments, we have implemented our approach in a computer program, MassXplorer. We demonstrate the application of MassXplorer to two model mixtures of known proteins and to a complex mixture of mouse kidney cortical extract. Comparison with another algorithm for ratio estimations demonstrates the increased precision and automation of MassXplorer.
Numerous metabolic pathways have been implicated in diabetes-induced renal injury, yet few studies have utilized unbiased systems biology approaches for mapping the interconnectivity of diabetes-dysregulated proteins that are involved. We utilized a global, quantitative, differential proteomic approach to identify a novel retinoic acid hub in renal cortical protein networks dysregulated by type 2 diabetes.
Shandong Province is located in the east of China. The province is characterized by robust economic development, with a rural population accounting for 56% of the total population. However, no data are available regarding temporal changes in the prevalence, awareness, treatment and control of hypertension among this population. Three independent cross-sectional surveys were performed in 1991 (n=8359), 2002 (n=18922) and 2007 (n=20167) in the rural area population, aged 35-74 years. The sampling included a survey on blood pressure and associated risk factors. Although the rate of smoking and alcohol consumption decreased significantly from 1991 to 2007, the prevalence of overweight and obesity increased, whereas that of high-strength physical activity decreased remarkably. In 1991, 2002 and 2007, the prevalence of prehypertension was 33.8, 61.5 and 54.6%, respectively. The prevalence of hypertension was 20.4, 24.5 and 30.6%, respectively. Overall, the rate of hypertension awareness, treatment and control showed a steady increase over the 16-year period, although the control rate of hypertension is still far from being satisfactory. In conclusion, among the Chinese rural population, the prevalence of prehypertension and hypertension increased significantly from 1991 to 2007. Public health programs are required to improve this situation in Chinese rural populations.
Although a science advisory recommending 12 months of dual antiplatelet therapy after drug-eluting stents implantation was published recently, the optimal duration of dual antiplatelet therapy has not yet been precisely determined.
A fiber-based biofilm reactor was developed using a laboratory-scale apparatus for treatment of nitrate-contaminated groundwater. Denitrification bacteria were inoculated by anaerobic sludge from a wastewater treatment plant. Nitrate removal efficiency, nitrite accumulation, COD and pH in the treated water were investigated under various conditions set by several parameters including hydraulic retention times (HRTs) (24, 20, 16, 12, 8, 4 and 2h), influent nitrate loading (around 50, 100 and 150 NO(3)(-)-N mg L(-1)), pH (5, 6, 7, 8, and 9) and ratios of carbon to nitrogen (C/N=3.00, 2.00, 1.50 1.25 and 1.00). The experimental results demonstrated that the optimum reaction parameters were pH 7-7.5,C/N=1.25 and HRT=8h, under which over 99% of NO(3)(-)-N was removed, almost no NO(2)(-)-N accumulated and COD was nearly zero in treated water when the concentration of NO(3)(-)-N was around 100.00 mg L(-1) in influent.
Economy has developed rapidly in China, and the clustering of cardiovascular risk factors in subjects increased remarkably over the past two decades. However, no data are available regarding the temporal prevalence of hyperuricemia and its correlates in this rapidly developing area, especially in the inland area. The cross-sectional survey was based on a random sample of 4,218 residents aged 35-64 years in the Jinan area. Hyperuricemia was defined as serum uric acid ? 416 ?mol/L in men and ? 357 ?mol/L in women. Subjects underwent physical examination and fasting blood testing. Complete data were available for analysis from 1,979 men and 2,062 women. The age-adjusted prevalence of hyperuricemia was 6.4 % for men and 2.1 % for women. The prevalence of hyperuricemia was greater in urban (6.7 %) than in rural areas (1.7 %) of Jinan city. Multivariate logistic regression models revealed hyperuricemia associated with hypertriglyceridemia [men: odds ratio (OR) = 6.101, 95 % confidence interval (CI) 4.064-9.159; women: OR = 7.103, 95 % CI 3.578-14.099] and high serum creatinine level (men: OR = 2.603, 95 % CI 1.602-4.230; women: OR = 5.237, 95 % CI 2.667-10.284). Hyperuricemia was also significantly associated with male sex, urban residence, hypertension, obesity, and hypercholesterolemia. Age (1-year increase) was negatively associated with hyperuricemia in men but positively associated with hyperuricemia in women. In conclusion, the prevalence of hyperuricemia is higher in urban than rural areas of Jinan, China. Male sex, urban residence, hypertension, obesity, hypercholesterolemia, hypertriglyceridemia, and high serum creatinine level contributed to hyperuricemia in this population.
We previously reported that the peroxisome proliferator-activated receptor ? (PPAR?) agonist rosiglitazone (RSG) improved hippocampus-dependent cognition in the Alzheimers disease (AD) mouse model, Tg2576. RSG had no effect on wild-type littermate cognitive performance. Since extracellular signal-regulated protein kinase mitogen-activated protein kinase (ERK MAPK) is required for many forms of learning and memory that are affected in AD, and since both PPAR? and ERK MAPK are key mediators of insulin signaling, the current study tested the hypothesis that RSG-mediated cognitive improvement induces a hippocampal PPAR? pattern of gene and protein expression that converges with the ERK MAPK signaling axis in Tg2576 AD mice. In the hippocampal PPAR? transcriptome, we found significant overlap between peroxisome proliferator response element-containing PPAR? target genes and ERK-regulated, cAMP response element-containing target genes. Within the Tg2576 dentate gyrus proteome, RSG induced proteins with structural, energy, biosynthesis and plasticity functions. Several of these proteins are known to be important for cognitive function and are also regulated by ERK MAPK. In addition, we found the RSG-mediated augmentation of PPAR? and ERK2 activity during Tg2576 cognitive enhancement was reversed when hippocampal PPAR? was pharmacologically antagonized, revealing a coordinate relationship between PPAR? transcriptional competency and phosphorylated ERK that is reciprocally affected in response to chronic activation, compared with acute inhibition, of PPAR?. We conclude that the hippocampal transcriptome and proteome induced by cognitive enhancement with RSG harnesses a dysregulated ERK MAPK signal transduction pathway to overcome AD-like cognitive deficits in Tg2576 mice. Thus, PPAR? represents a signaling system that is not crucial for normal cognition yet can intercede to restore neural networks compromised by AD.
A novel ceramic adsorbent was developed to adsorb ammonium from high concentration ammonium contaminated wastewater. Typical gardening cultivation mediums in Japan-Kanuma clay and Akadama clay were used to synthesize the ceramic adsorbent. Static batch experiments were conducted to investigate the effect of various parameters such as contact time, initial ammonium concentration, adsorbent dosage, and competing cations during the ammonium adsorption process. The results revealed that the Freundlich isotherm model fitted better with the adsorption process than the Langmuir model, and the adsorption process was well described by pseudo-second-order kinetic model. The maximum nitrogen adsorption capacity of the ceramic adsorbent was 75.5 mg g(-1) at an initial NH(4)(+)-N concentration of 10,000 mg L(-1), dosage of 20 g L(-1), and contact time of 480 min. Results demonstrated that the low-cost ceramic adsorbent directly used as nitrogen fertilizer was feasible for its high ammonium nitrogen content, nontoxic effect on the environment and excellent soil properties.
The authors retrospectively examined data from 3 surveys on hypertension according to the 2010 Chinese Guidelines for the Management of Hypertension. These surveys were conducted in 1991, 1999, and 2007, and included 85,371 individuals 18 years and older who were living in Shandong Province, China. Age-standardized prevalent hypertension increased from 15.6% in 1991 to 17.3% in 1999 and 32.7% in 2007 (both P<.0001). The ascending prevalence can be partially explained by increasing body weight. Among individuals with hypertension, awareness increased significantly from 27.8% in 1991 to 39.1% in 1999 and 49.2% in 2007. The proportion of pharmacologic treatment also considerably improved, with the estimate of 12.9%, 28.1%, and 43.3% in the 3 surveys, respectively. Hypertension control increased from 3.0% to 4.4% to 7.1% in the past 20 years. The upward trend in blood pressure control was mostly attributable to a rise among men and persons at middle age. This study suggests that the prevalence of hypertension increased in the Shandong population from 1991 to 2007. Although substantially improved, control rates were still unacceptably low. Comprehensive strategies are urgently required to put into practice for the management of hypertension in Shandong Province, China.
Traumatic brain injury (TBI) is a complex and common problem resulting in the loss of cognitive function. In order to build a comprehensive knowledge base of the proteins that underlie these cognitive deficits, we employed unbiased quantitative mass spectrometry, proteomics, and bioinformatics to identify and quantify dysregulated proteins in the CA3 subregion of the hippocampus in the fluid percussion model of TBI in rats. Using stable isotope 18O-water differential labeling and multidimensional tandem liquid chromatography (LC)-MS/MS with high stringency statistical analyses and filtering, we identified and quantified 1002 common proteins, with 124 increased and 76 decreased. The ingenuity pathway analysis (IPA) bioinformatics tool identified that TBI had profound effects on downregulating global energy metabolism, including glycolysis, the Krebs cycle, and oxidative phosphorylation, as well as cellular structure and function. Widespread upregulation of actin-related cytoskeletal dynamics was also found. IPA indicated a common integrative signaling node, calcineurin B1 (CANB1, CaNB?, or PPP3R1), which was downregulated by TBI. Western blotting confirmed that the calcineurin regulatory subunit, CANB1, and its catalytic binding partner PP2BA, were decreased without changes in other calcineurin subunits. CANB1 plays a critical role in downregulated networks of calcium signaling and homeostasis through calmodulin and calmodulin-dependent kinase II to highly interconnected structural networks dominated by tubulins. This large-scale knowledge base lays the foundation for the identification of novel therapeutic targets for cognitive rescue in TBI.
Junín virus (JUNV), an arenavirus, is the causative agent of Argentine hemorrhagic fever, an infectious human disease with 15-30% case fatality. The pathogenesis of AHF is still not well understood. Elevated levels of interferon and cytokines are reported in AHF patients, which might be correlated to the severity of the disease. However the innate immune response to JUNV infection has not been well evaluated. Previous studies have suggested that the virulent strain of JUNV does not induce IFN in human macrophages and monocytes, whereas the attenuated strain of JUNV was found to induce IFN response in murine macrophages via the TLR-2 signaling pathway. In this study, we investigated the interaction between JUNV and IFN pathway in human epithelial cells highly permissive to JUNV infection. We have determined the expression pattern of interferon-stimulated genes (ISGs) and IFN-? at both mRNA and protein levels during JUNV infection. Our results clearly indicate that JUNV infection activates the type I IFN response. STAT1 phosphorylation, a downstream marker of activation of IFN signaling pathway, was readily detected in JUNV infected IFN-competent cells. Our studies also demonstrated for the first time that RIG-I was required for IFN production during JUNV infection. IFN activation was detected during infection by either the virulent or attenuated vaccine strain of JUNV. Curiously, both virus strains were relatively insensitive to human IFN treatment. Our studies collectively indicated that JUNV infection could induce host type I IFN response and provided new insights into the interaction between JUNV and host innate immune system, which might be important in future studies on vaccine development and antiviral treatment.
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