Fabrication of a Near-Infrared-Emissive Probe for Detecting Dipeptidyl Peptidase 4 in the Liver of Diabetic Mice and Clinical Serum.

Dipeptidyl peptidase 4 (DPP4) plays a key role in glucose metabolism, which has been a close target for diabetes pathology and treatment. It is significant for the evaluation of cellular DPP4 activity in various biological systems. Fluorescence imaging technology is currently a popular method for detecting enzymes in living cells due to its advantages of high selectivity, high sensitivity, high spatiotemporal resolution, and real-time visualization. Herein, a near-infrared (NIR)-emissive probe NEDP with a large Stokes shift (153 nm) was developed for the assay of DPP4 activity. Upon addition of DPP4, NEDP can emit a significant turn-on NIR fluorescence signal (673 nm) with high sensitivity and specificity. Moreover, NEDP can successfully be used for imaging of intracellular DPP4, confirming the regulation of DPP4 expression in hyperglucose and its treatment in living cells. Most importantly, NEDP can not only monitor the changes of DPP4 in vivo but also show that DPP4 in diabetes is mainly up-regulated in the liver, and the level of DPP4 is positively correlated with the pathological damage of the liver. In addition, NEDP can identify the serum of diabetic patients from healthy people through the fluorescence response to DPP4. These results demonstrated that the designed probe NEDP provides a prospective visual tool to explore the relationship between DPP4 and diabetes and would be applied for detecting serum of diabetes in the clinic.

Serum soluble alpha-klotho klotho and cognitive functioning in older adults aged 60 and 79: an analysis of cross-sectional data of the National Health and Nutrition Examination Survey 2011 to 2014.

OBJECTIVES: Klotho, consisting of membrane klotho and soluble alpha-klotho, is found to be associated with better cognitive outcomes in small samples of the aged population. We aimed to examine the association of serum soluble alpha-klotho with cognitive functioning among older adults using a nationally representative sample of U.S. older adults. METHOD: A total of 2,173 U.S. older adults aged 60-79 years in the National Health and Nutrition Examination Survey from 2011 to 2014 were included in this cross-sectional analysis. Serum soluble alpha-klotho was measured in the laboratory and analyzed with an ELISA kit. Cognitive function was measured using the Consortium to Establish a Registry for Alzheimer's Disease Word Learning subtest (CERAD-WL) immediate and delayed memory, the Animal fluency test (AFT), and the Digit Symbol Substitution Test (DSST). Test-specific and global cognition z-scores were calculated based on sample means and standard deviations. Multivariable linear regression models were applied to examine the association of quartiles and continuous value of serum soluble alpha-klotho with test-specific and global cognition z-scores. Subgroup analysis was conducted by sex. The following covariates were included in the analysis- age, sex, race/ethnicity, education, depressive symptoms, smoking status, body mass index (BMI), physical activity, stroke, prevalent coronary heart disease, total cholesterol, and systolic blood pressure. All the information was self-reported or obtained from health exams. RESULTS: Serum soluble alpha-klotho level in the lowest quartile was associated with lower z-scores for DSST (beta [ß] =-0.13, 95% confidence interval [CI]: -0.25, -0.01). For subgroup analysis, serum soluble alpha-klotho level in the lowest quartile was associated with lower z-scores for DSST (ß=-0.16, 95% CI: -0.32, -0.003) and global cognition (ß=-0.14, 95% CI: -0.28, -0.01) among female participants. No association was found between continuous serum soluble alpha-klotho and cognitive functioning among the participants. CONCLUSIONS: Lower serum soluble alpha-klotho quartile was associated with poorer cognitive functioning among older women. Future studies are expected to examine the longitudinal association between klotho levels and cognitive outcomes.

A single mitochondria-targetable fluorescent probe for visualizing cysteine and glutathione in ferroptosis of myocardial ischemia/reperfusion injury.

Ferroptosis plays an important role in the early stage of myocardial ischemia/reperfusion (MI/R) injury, which is closely associated with the antioxidant damage of mitochondrial cysteine (Cys)/glutathione (GSH)/glutathione peroxidase 4 (GPX4) axis. Visualization of Cys and GSH in mitochondria is meaningful to value ferroptosis and further contributes to understanding and preventing MI/R injury. Herein a mitochondria-targetable thiols fluorescent probe (MTTP) was designed and synthesized based on sulfonyl benzoxadiazole (SBD) chromophore with a triphenylphosphine unit as the mitochondria-targeted functional group. Cys and GSH can be differentiated by MTTP with two distinguishable emission bands (583 nm and 520 nm) through the controllable aromatic substitution-rearrangement reaction. Importantly, MTTP is capable of monitoring ferroptosis and its inhibition by measuring mitochondrial Cys and GSH. MTTP was also employed to non-invasively detect ferroptosis during oxygen and glucose deprivation/reoxygenation (OGD/R)-induced MI/R injury in H9C2 cells. In a word, MTTP provides a visual tool that can simultaneously detect Cys and GSH to monitor ferroptosis processes during MI/R injury, which helps for more deeper understanding of the role of ferroptosis in MI/R injury-related diseases.

Association between seropositivity for toxocariasis and cognitive functioning in older adults: an analysis of cross-sectional data from the US National Health and Nutrition Examination Survey (NHANES), 2011-2014.

OBJECTIVES: This study sought to examine the relationship between seropositivity for toxocariasis and cognitive functioning in a nationally representative sample of US older adults. DESIGN: A cross-sectional study. SETTING: National Health and Nutrition Examination Survey (NHANES) data collection took place in the US at participants' homes and mobile examination centres with specialised equipment. PARTICIPANTS: The study population consisted of 3188 community-dwelling US older adults aged 60 and above from the NHANES 2011 to 2014. OUTCOME MEASURES: IgG antibody against Toxocara spp was tested by a Luminex assay using recombinant rTc-CTL-1 antigen. A value >23.1 mean fluorescence intensity (MFI) indicated positive for toxocariasis and a value ≤23.1 MFI as negative for toxocariasis. The Consortium to Establish a Registry for Alzheimer's Disease Word Learning subtest immediate and delayed memory, the Animal Fluency test and the Digit Symbol Substitution Test (DSST) were used to assess cognitive functioning. Cognitive test-specific and global cognitive z scores were computed using sample means and SD. RESULTS: The study population consisted of 3188 participants who represented a total of 111 896 309 civilian citizens in the USA. The mean age of the participants was 69.6 years (standard deviation 6.8). The prevalence of toxocariasis in this population was 7.3% (95% confidence interval [CI] 6.1% to 8.5%). The survey-weighted linear regression model showed that compared with participants who were toxocariasis seronegative, those who were seropositive had lower DSST z score (beta [ß] = -0.12, 95% CI -0.22 to -0.01) and global cognition z score (ß=-0.11, 95% CI -0.22 to -0.01), after controlling for age, sex, race/ethnicity, education, depressive symptoms, smoking status, body mass index, prevalent coronary heart disease, prevalent stroke, and systolic blood pressure, physical activity, and total cholesterol. CONCLUSIONS: In our study, seropositive toxocariasis was independently and significantly associated with worse working memory, sustained attention, processing speed and global cognition in older adults. If this association is causal, public health measures to prevent human toxocariasis might help protect older adults' cognitive function.

Advances in Multifunctional Chemotherapeutic Prodrugs for Near-infrared Fluorescence Imaging-guided Therapy.

Conventional chemotherapy (CT) is associated with severe side effects and inducible resistance, making it difficult to meet clinical requirements, forcing the development of new multifunctional prodrugs for precision medicine. In recent decades, researchers and clinicians have focused on developing of multifunctional chemotherapeutic prodrugs with tumor-targeting capability, activatable and traceable chemotherapeutic activity, as a powerful tool to improve theranostic outcomes in cancer treatment. The conjugates of near-infrared (NIR) organic fluorophores and chemotherapy reagents create an exciting avenue for real-time monitoring of drug delivery and distribution, as well as the combination of chemotherapy and photodynamic therapy (PDT). Therefore, there are great opportunities for researchers to conceive and exploit multifunctional prodrugs that can visualize chemo-drugs release and tumor treatment in vivo. In this review, the design strategy and the recent progress of multifunctional organic chemotherapeutic prodrugs for activating NIR fluorescence imaging-guided therapy are described and discussed in detail. Finally, the prospects and challenges of multifunctional chemotherapeutic prodrugs for NIR fluorescence imaging-guided therapy are provided.

Treatment-Interval Changes in Serum Levels of Albumin and Histidine Correlated with Treatment Interruption in Patients with Locally Advanced Head and Neck Squamous Cell Carcinoma Completing Chemoradiotherapy under Recommended Calorie and Protein Provision.

We investigated risk factors for treatment interruption (TI) in patients with locally advanced head and neck squamous-cell carcinoma (LAHNSCC) following concurrent chemoradiotherapy (CCRT), under the provision of recommended calorie and protein intake; we also evaluated the associations between clinicopathological variables, calorie and protein supply, nutrition-inflammation biomarkers (NIBs), total body composition change (TBC), and a four-serum-amino-acid metabolite panel (histidine, leucine, ornithine, and phenylalanine) among these patients. Patients with LAHNSCC who completed the entire planned CCRT course and received at least 25 kcal/kg/day and 1 g of protein/kg/day during CCRT were prospectively recruited. Clinicopathological variables, anthropometric data, blood NIBs, CCRT-related factors, TBC data, and metabolite panels before and after treatment were collected; 44 patients with LAHNSCC were enrolled. Nine patients (20.4%) experienced TIs. Patients with TIs experienced greater reductions in hemoglobin, serum levels of albumin, uric acid, histidine, and appendicular skeletal mass, and suffered from more grade 3/4 toxicities than those with no TI. Neither increased daily calorie supply (≥30 kcal/kg/day) nor feeding tube placement was correlated with TI. Multivariate analysis showed that treatment-interval changes in serum albumin and histidine levels, but not treatment toxicity, were independently associated with TI. Thus, changes in serum levels of albumin and histidine over the treatment course could cause TI in patients with LAHNSCC following CCRT.

Lineage tracing of direct astrocyte-to-neuron conversion in the mouse cortex.

Regenerating functional new neurons in the adult mammalian central nervous system has been proven to be very challenging due to the inability of neurons to divide and repopulate themselves after neuronal loss. Glial cells, on the other hand, can divide and repopulate themselves under injury or diseased conditions. We have previously reported that ectopic expression of NeuroD1 in dividing glial cells can directly convert them into neurons. Here, using astrocytic lineage-tracing reporter mice (Aldh1l1-CreERT2 mice crossing with Ai14 mice), we demonstrate that lineage-traced astrocytes can be successfully converted into NeuN-positive neurons after expressing NeuroD1 through adeno-associated viruses. Retroviral expression of NeuroD1 further confirms that dividing glial cells can be converted into neurons. Importantly, we demonstrate that for in vivo cell conversion study, using a safe level of adeno-associated virus dosage (1010-1012 gc/mL, 1 µL) in the rodent brain is critical to avoid artifacts caused by toxic dosage, such as that used in a recent bioRxiv study (2 × 1013 gc/mL, 1 µL, mouse cortex). For therapeutic purpose under injury or diseased conditions, or for non-human primate studies, adeno-associated virus dosage needs to be optimized through a series of dose-finding experiments. Moreover, for future in vivo glia-to-neuron conversion studies, we recommend that the adeno-associated virus results are further verified with retroviruses that mainly express transgenes in dividing glial cells in order to draw solid conclusions. The study was approved by the Laboratory Animal Ethics Committee of Jinan University, China (approval No. IACUC-20180330-06) on March 30, 2018.

Independent and synergistic effects of pain, insomnia, and depression on falls among older adults: a longitudinal study.

BACKGROUND: Few studies have examined the relationship between falls and pain, insomnia and depressive symptoms which are common and risk factors in older adults. We aimed to examine the independent and synergistic effects of these risk factors on future falls among older adults. METHODS: We used data of 2558 community-dwelling older adults from 2011 (Y1) to 2015 (Y5) of the National Health and Aging Trends Study (NHATS). Pain was determined by whether participants reported bothersome pain in the last month. Insomnia was assessed by two questions about how often the participants had trouble falling asleep and maintaining sleep. Depressive symptoms were assessed by Patient Health Questionnaire-2. Generalized estimation equation (GEE) models were used to examine the independent effects of pain, insomnia and depressive symptoms at prior-wave (period y-1) on falls at current wave (period y) adjusting for covariates (age, sex, education, race/ethnicity, living arrangement, BMI, smoking, vigorous activities, number of chronic illnesses and hospitalization). The significance of the three-way interaction of these factors (pain*insomnia*depression) was tested using the aforementioned GEE models to determine their synergistic effects on falls. RESULTS: Overall, the participants were mainly 65-79 years old (68%), female (57%) and non-Hispanic White (70%). At Y1, 50.0% of the participants reported pain, 22.6% reported insomnia and 9.9% reported depressive symptoms. The incidence of falls from Y2 to Y5 was 22.4, 26.0, 28.3, and 28.9%, respectively. Participants with pain (Odds ratio [OR], 95% confidence interval [CI] = 1.36, 1.23-1.50) and depressive symptoms (OR, 95% CI = 1.43, 1.23-1.67) had high rates of falling adjusting for covariates. After further adjustment for insomnia and depressive symptoms, pain independently predicted falls (OR, 95% CI = 1.36, 1.22-1.51). Depressive symptoms also independently predicted falls after further adjusting for pain and insomnia (OR, 95% CI = 1.40, 1.20-1.63). After adjusting for pain and depression, the independent effects of insomnia were not significant. None of the interaction terms of the three risk factors were significant, suggesting an absence of their synergistic effects. CONCLUSIONS: Pain and depressive symptoms independently predict falls, but synergistic effects seem absent. Further research is needed to develop effective strategies for reducing falls in older adults, particularly with pain and depressive symptoms.

Gene therapy conversion of striatal astrocytes into GABAergic neurons in mouse models of Huntington's disease.

Huntington's disease (HD) is caused by Huntingtin (Htt) gene mutation resulting in the loss of striatal GABAergic neurons and motor functional deficits. We report here an in vivo cell conversion technology to reprogram striatal astrocytes into GABAergic neurons in both R6/2 and YAC128 HD mouse models through AAV-mediated ectopic expression of NeuroD1 and Dlx2 transcription factors. We found that the astrocyte-to-neuron (AtN) conversion rate reached 80% in the striatum and >50% of the converted neurons were DARPP32+ medium spiny neurons. The striatal astrocyte-converted neurons showed action potentials and synaptic events, and projected their axons to the targeted globus pallidus and substantia nigra in a time-dependent manner. Behavioral analyses found that NeuroD1 and Dlx2-treated R6/2 mice showed a significant extension of life span and improvement of motor functions. This study demonstrates that in vivo AtN conversion may be a disease-modifying gene therapy to treat HD and other neurodegenerative disorders.

Identifying hepatocellular carcinoma-related hub genes by bioinformatics analysis and CYP2C8 is a potential prognostic biomarker.

Hepatocellular carcinoma (HCC) is one type of the most common malignancies. However, the underlying molecular mechanisms involved in the development of HCC remain unknown. To identify the candidate genes in the progression of HCC, gene expression profiles GSE14520, GSE54236, GSE57957 and GSE64041 were downloaded from the Gene Expression Omnibus database (GEO). A total of 405 tumor and 399 para-carcinoma samples from patients with HCC were examined to identify the differentially expressed genes (DEGs), followed by function enrichment analyses including Gene Ontology (GO) functions and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. A total of 78 DEGs were screened, including 62 downregulated genes and 16 upregulated genes. Subsequently, the protein-protein interaction network (PPI) was constructed using the Search Tool for Retrieval of Interacting Genes (STRING) database. The module analysis and Hub genes validation were performed using Cytoscape software. Hierarchical clustering of hub genes was evaluated using UCSC Cancer Genomics Browser. Survival analyses of Hub genes were performed using Kaplan Meier Plotter database. Genes specifically expressed in the liver were analyzed using GENEVESTIGATOR database. CYP2C8 was identified as one of the most promising molecules among all the candidate genes. The expression profile of CYP2C8 in HCC was analyzed using ONCOMINE and UALCAN database. The expression levels of CYP2C8 in HCC samples and hepatoma cells were verified using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry analysis. In summary, DEGs and hub genes were identified in the present study, which provides novel insight on the development of HCC. CYP2C8 was downregulated in HCC and could be a potential prognostic biomarker.

Interleukin 10 Gene-Modified Bone Marrow-Derived Dendritic Cells Attenuate Liver Fibrosis in Mice by Inducing Regulatory T Cells and Inhibiting the TGF-ß/Smad Signaling Pathway.

AIM: To explore the therapeutic effects and mechanisms of interleukin 10 gene-modified bone marrow-derived dendritic cells (DC-IL10) on liver fibrosis. METHODS: In vitro, BMDCs were transfected with lentiviral-interleukin 10-GFP (LV-IL10-GFP) at the MOI of 1 : 40. Then, the phenotype (MHCII, CD80, and CD86) and allo-stimulatory ability of DC-IL10 were identified by flow cytometry, and the levels of IL-10 and IL-12 (p70) secreted into the culture supernatants were quantified by ELISA. In vivo, DC-IL10 was injected into mice with CCl4-induced liver fibrosis through the tail vein. Lymphocytes were isolated to investigate the differentiation of T cells, and serum and liver tissue were collected for biochemical, cytokine, histopathologic, immune-histochemical, and Western blot analyzes. RESULTS: In vitro, the expressions of MHCII, CD80, and CD86 in DC-IL10 were significantly suppressed, allogeneic CD4+T cells incubated with DC-IL10 showed a lower proliferative response, and the levels of IL-10 and IL-12 (p70) secreted into the DC-IL10 culture supernatants were significantly increased and decreased, respectively. In vivo, regulatory T cells (Tregs) were significantly increased, while ALT, AST, and inflammatory cytokines were significantly reduced in the DC-IL10 treatment group, and the degree of hepatic fibrosis was obviously reversed. The TGF-ß/smad pathway was inhibited following DC-IL10 treatment compared to the liver fibrosis group. CONCLUSION: IL-10 genetic modification of BMDCs may maintain DC in the state of tolerance and allow DC to induce T cell hyporesponsiveness or tolerance. DC-IL10 suppressed liver fibrosis by inducing Treg production and inhibiting the TGF-ß/smad signaling pathway.

Perchlorate Removal in Microbial Electrochemical Systems With Iron/Carbon Electrodes.

Perchlorate removal was tested in the cathode chamber of microbial electrochemical systems (MESs). Dual-chambers MESs were constructed and operated in batch mode with four kinds of cathode materials including Fe/C particles (Fe/C), zero valent iron particles (ZVI), blank carbon felt (CF), and active carbon (AC). Without external energy supply or perchlorate-reducing microbial pre-enrichment, perchlorate ( ClO 4 - ) removal could be achieved in the cathode chambers of MESs at different efficiencies. The highest ClO 4 - removal rates in these reactors were 18.96 (Fe/C, 100 Ω, 2 days), 15.84 (ZVI, 100 Ω, 2 days), 14.37 (CF, 100 Ω, 3 days), and 19.78 mg/L/day (AC, 100 Ω, 2 days). ClO 4 - degradation products were mainly Cl- and ClO 3 - , and the total chlorine in the products was lower than the theoretical input. The non-conservation of the total chlorine may be caused by the adsorption and co-precipitation related to the electrode materials. Coulombs and coulombic efficiency calculation showed that electron provided by MESs was partially responsible for ClO 4 - reduction, for the Fe/C cathode reactors, about a quarter of electron was provided by MESs.

Estimating systemic fibrosis by combining galectin-3 and ST2 provides powerful risk stratification value for patients after acute decompensated heart failure.

BACKGROUND: Two fibrosis biomarkers, galectin-3 (Gal-3) and suppression of tumorigenicity 2 (ST2), provide prognostic value additive to natriuretic peptides and traditional risk factors in patients with heart failure (HF). However, it is to be investigated whether their combined measurement before discharge provides incremental risk stratification for patients after acute HF. METHODS: A total of 344 patients with acute HF were analyzed with Gal-3, and ST2 measured. Patients were prospectively followed for 3.7 ± 1.3 years for deaths, and composite events (death/HF-related re-hospitalizations). RESULTS: The levels of Gal-3 and ST2 were only slightly related (r = 0.20, p < 0.001). The medians of Gal-3 and ST2 were 18 ng/mL and 32.4 ng/mL, respectively. These biomarkers compensated each other and characterized patients with different risk factors. According to the cutoff at median values, patients were separated into four subgroups based on high and low Gal-3 (HG and LG, respectively) and ST2 levels (HS and LS, respectively). Kaplan-Meier survival curves showed that HGHS powerfully identified patients at risk of mortality (Log rank = 21.27, p < 0.001). In multivariable analysis, combined log(Gal-3) and log(ST2) was an in-dependent predictor. For composite events, Kaplan-Meier survival curves showed a lower event- -free survival rate in the HGHS subgroup compared to others (Log rank = 34.62, p < 0.001; HGHS vs. HGLS, Log rank = 4.00, p = 0.045). In multivariable analysis, combined log(Gal-3) and log(ST2) was also an independent predictor. CONCLUSIONS: Combination of biomarkers involving heterogeneous fibrosis pathways may identify patients with high systemic fibrosis, providing powerful risk stratification value.

Effect of multidisciplinary disease management for hospitalized heart failure under a national health insurance programme.

AIM: Multidisciplinary disease management programmes (MDPs) for heart failure have been shown to be effective in Western countries. However, it is not known whether they improve outcomes in a high population density country with a national health insurance programme. METHODS: In total, 349 patients hospitalized because of heart failure were randomized into control and MDP groups. All-cause death and re-hospitalization related to heart failure were analyzed. The median follow-up period was approximately 2 years. RESULTS: Mean patient age was 60 years; 31% were women; and 50% of patients had coronary artery disease. MDP was associated with fewer all-cause deaths [hazard ratio (HR) = 0.49, 95% confidence interval (CI) = 0.27-0.91, P = 0.02] and heart failure-related re-hospitalizations (HR = 0.44, 95% CI = 0.25-0.77, P = 0.004). MDP was still associated with better outcomes for all-cause death (HR = 0.53, 95% CI = 0.29-0.98, P = 0.04) and heart failure-related re-hospitalization (HR = 0.46, 95% CI = 0.26-0.81, P = 0.007), after adjusting for age, diuretics, diabetes mellitus, chronic kidney disease, hypertension, sodium, and albumin. However, MDPs' effect on all-cause mortality and heart failure-related re-hospitalization was significantly attenuated after adjusting for angiotensin-converting enzyme inhibitors/angiotensin II receptor blockers or ß-blockers. A stratified analysis showed that MDP combined with guideline-based medication had synergistic effects. CONCLUSIONS: MDP is effective in lowering all-cause mortality and re-hospitalization rates related to heart failure under a national health insurance programme. MDP synergistically improves the effectiveness of guidelines-based medications for heart failure.

Ultrasound elastography in the differential diagnosis of benign and malignant cervical lesions.

OBJECTIVES: This study aimed to evaluate the clinical value of ultrasound elastography in the differential diagnosis of benign and malignant cervical lesions and to compare the accuracy of the elasticity score and strain ratio in differentiating cervical lesions. METHODS: B-mode sonography and ultrasound elastography were performed on 84 cervical lesions (40 benign and 44 malignant) in 84 patients. All of the images were obtained transvaginally. The elasticity score was determined by a 5-point scoring method. Calculation of the strain ratio was based on a comparison of the average strain measured in the lesion with the adjacent tissue of the same depth, size, and shape. The findings were compared with histopathologic results. With the use of receiver operating characteristic curves, the diagnostic value of the elasticity score and strain ratio methods was determined. RESULTS: The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of the elasticity score in the differential diagnosis of cervical lesions were 81.8%, 85.0%, 83.3%, 85.7%, and 81.0%, respectively, whereas those of the strain ratio were 90.9%, 90.0%, 90.5%, 90.9%, and 90.0%. A strain ratio cutoff value of 4.525 was used as a standard to distinguish benign from malignant lesions. The strain ratio values of malignant lesions were much higher than those of benign lesions (range, 4.85-8.91 versus 0.62-4.50). The differences were statistically significant (P < .01). CONCLUSIONS: Ultrasound elastography is a promising technique that is easy and rapid to perform and can help identify cervical lesions that are likely to be malignant. It is obvious that the strain ratio yielded better results than the elasticity score. Both methods are semiquantitative, but quantification of the strain ratio is finer than that of the elasticity score.

Serum interleukin-6 levels, not genotype, correlate with coronary plaque complexity.

An increased serum interleukin-6 (IL-6) level is associated with an increased risk of cardiovascular events in healthy subjects. However, it is unknown whether the level of serum IL-6 or genetic IL-6 polymorphism is correlated with the complexity of coronary plaque in patients with stable coronary artery disease (CAD). Patients with stable CAD (n = 135) were divided into 3 groups: insignificant coronary plaque (n = 77), simple coronary plaque (n = 15), and complex coronary plaque (n = 43). IL-6-174G > C polymorphism and serum levels of IL-6 and C-reactive protein (CRP) were investigated. No significant difference in the distribution of IL-6 genotypes was found among the groups. The presence of complex coronary plaque was associated with higher serum concentrations of IL-6 (P = 0.026) and CRP (P < 0.0001). To predict the presence of complex lesions, IL-6 > 5.8 ng/L and CRP > 2.6 mg/L had sensitivities of 86% and 74%, and specificities of 61% and 62%, respectively. By multivariate analysis, IL-6 > 5.8 ng/L and CRP > 2.6 mg/L were independently related to the presence of complex coronary plaque (P = 0.0002 and 0.004, respectively). IL-6 > 5.8 ng/L and CRP > 2.6 mg/L were associated with a 4.5-fold increase in the odds of having complex coronary plaque (P < 0.005). A simple measurement of the serum IL-6 level in patients with CAD can potentially identify subjects with complex coronary lesions and provide the option of aggressive medical strategies in a clinical setting.

Effects of resveratrol, a grape polyphenol, on catecholamine secretion and synthesis in cultured bovine adrenal medullary cells.

We report the effects of resveratrol, a polyphenol found in the skins of red grapes, on catecholamine secretion and synthesis in cultured bovine adrenal medullary cells. Resveratrol suppressed catecholamine secretion and (22)Na(+) and (45)Ca(2+) influx induced by acetylcholine, an agonist of nicotinic acetylcholine receptors, in a concentration-dependent manner (IC(50)=20.4, 11.0, and 62.8 microM, respectively). Resveratrol also inhibited catecholamine secretion induced by veratridine, an activator of voltage-dependent Na(+) channels, and 56 mM K(+), an activator of voltage-dependent Ca(2+) channels, at concentrations similar to those for (45)Ca(2+) influx. Resveratrol directly inhibited the current evoked by acetylcholine in Xenopus oocytes expressing alpha3beta4 neuronal nicotinic acetylcholine receptors (IC(50)=25.9 microM). Furthermore, resveratrol (IC(50)=5.32 microM) attenuated (14)C-catecholamine synthesis induced by acetylcholine. The present findings suggest that resveratrol inhibits acetylcholine-induced catecholamine secretion and synthesis through suppressing ion influx in cultured bovine adrenal medullary cells.

Dual effects of daidzein, a soy isoflavone, on catecholamine synthesis and secretion in cultured bovine adrenal medullary cells.

We recently demonstrated the occurrence and functional roles of plasma membrane estrogen receptors in cultured bovine adrenal medullary cells. Here we report the effects of daidzein, a phytoestrogen of soybeans, on catecholamine synthesis and secretion in the cells. Incubation of cells with daidzein for 20 min increased the synthesis of (14)C-catecholamines from [(14)C]tyrosine but not [(14)C]dihydroxyphenylalanine, in a concentration-dependent manner (10-1000 nm). The stimulatory effect of daidzein on (14)C-catecholamine synthesis was not inhibited by ICI182,780, a classical estrogen receptor inhibitor. Acetylcholine, a physiological secretagogue, stimulated the synthesis of (14)C-catecholamines, which was suppressed by daidzein at 1 mum. Daidzein at high concentrations (1-100 microm) suppressed catecholamine secretion induced by acetylcholine. Furthermore, daidzein (10-1000 nm) inhibited the specific binding of [(3)H]17beta-estradiol to plasma membranes isolated from bovine adrenal medulla. The present findings suggest that daidzein at low concentrations stimulates catecholamine synthesis through plasma membrane estrogen receptors but at high concentrations inhibits catecholamine synthesis and secretion induced by acetylcholine in bovine adrenal medulla. The latter effect of daidzein may be a beneficial action on the cardiovascular system.

Stimulation of catecholamine synthesis through unique estrogen receptors in the bovine adrenomedullary plasma membrane by 17beta-estradiol.

Incubation of cultured bovine adrenal medullary cells with 17beta-estradiol (E(2)) (0.3-100nM) or membrane-impermeable E(2)-bovine serum albumin (100nM) acutely increased (14)C-catecholamine synthesis from [(14)C]tyrosine. The stimulatory effect of E(2) was not inhibited by ICI182,780, a nuclear estrogen receptor inhibitor. E(2) also increased tyrosine hydroxylase activity and p44/42MAPK phosphorylation, the former of which was attenuated by U0126, an inhibitor of p44/42MAPK kinase. The plasma membrane isolated from the gland showed two classes of specific binding sites of [(3)H]E(2) with apparent K(d)s of 3.2 and 106nM, and B(max)s of 0.44 and 8.5pmol/mg protein, respectively. The high-affinity binding of [(3)H]E(2) was most strongly inhibited by E(2) and phytoestrogens, and to lesser extents by other steroid hormones, while it was enhanced by ICI182,780 and environmental estrogenic pollutants. These findings suggest that E(2) acutely stimulates catecholamine synthesis via activation of p44/42MAPK through unique estrogen receptors in the plasma membrane of bovine adrenal medulla.