RNAs often exhibit a high degree of conformational dynamics and heterogeneity, leading to a rugged energy landscape. However, the roles of conformational heterogeneity and rapid dynamics in molecular recognition or RNA function have not been extensively elucidated. Ultrafast time-resolved fluorescence spectroscopic experiments were used here to probe picosecond dynamics of the theophylline-binding RNA aptamer. These studies showed that multiple conformations are populated in the free RNA indicating that this aptamer employs a conformational capture mechanism for ligand binding. The base on residue 27 in an internal loop exists in at least three conformational states in the free RNA, including binding competent and incompetent states that have distinct fluorescence decay signatures indicating different base stacking interactions. Picosecond dynamics were also detected by anisotropy experiments, where these motions indicate additional dynamics for base 27. The picosecond data show that theophylline binding shifts the equilibrium for conformations of base 27 from primarily stacked in the free RNA to mostly unstacked in the RNA-theophylline complex, as observed in the previous NMR structure. In contrast, base 10 in a second internal loop is mostly pre-organized in the free RNA, consistent with it being stacked between G11 and G25, as is observed in the bound state. Picosecond dynamics were also measured on a modified aptamer that binds with higher affinity to 3-methylxanthine than theophylline. The modified aptamer shows less heterogeneity in the aptamer-3-methylxanthine complex than what is observed in the theophylline aptamer-theophylline complex.

OBJECTIVE: To study the components in aerial part of Siegesbeckia pubescens. METHOD: The compounds were isolated and purified by silica gel, sephadex LH-20 and other column chromatography. Structures were elucidated by spectroscopic methods. RESULT: Four compounds were isolated from S. pubescens and were characterized as dimethyl-21-ethenetylene-darutigenol-3-O-beta-D-glucopyranosid (1), darutigenol (2), darutoside (3), stigmaster-3-O-beta-D-glucopyranosid (4). CONCLUSION: Compound 1 is a new compound.

BACKGROUND: Pulmonary vascular diseases are increasingly recognised as important clinical conditions. Pulmonary hypertension associated with a range of aetiologies is difficult to treat and associated with progressive morbidity and mortality. Current therapies for pulmonary hypertension include phosphodiesterase type 5 inhibitors, endothelin receptor antagonists, or prostacyclin mimetics. However, none of these provide a cure and the clinical benefits of these drugs individually decline over time. There is, therefore, an urgent need to identify new treatment strategies for pulmonary hypertension. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that the PPARbeta/delta agonist GW0742 induces vasorelaxation in systemic and pulmonary vessels. Using tissue from genetically modified mice, we show that the dilator effects of GW0742 are independent of the target receptor PPARbeta/delta or cell surface prostacyclin (IP) receptors. In aortic tissue, vascular relaxant effects of GW0742 were not associated with increases in cGMP, cAMP or hyperpolarisation, but were attributed to inhibition of RhoA activity. In a rat model of hypoxia-induced pulmonary hypertension, daily oral dosing of animals with GW0742 (30 mg/kg) for 3 weeks significantly reduced the associated right heart hypertrophy and right ventricular systolic pressure. GW0742 had no effect on vascular remodelling induced by hypoxia in this model. CONCLUSIONS/SIGNIFICANCE: These observations are the first to show a therapeutic benefit of 'PPARbeta/delta' agonists in experimental pulmonary arterial hypertension and provide pre-clinical evidence to favour clinical trials in man.

Ubiquitin ligase Smurf1-deficient mice develop an increased bone mass phenotype in an age-dependent manner. It was reported that such a bone mass increase is related to enhanced activities of differentiated osteoblasts. Though osteoblasts are of mesenchymal stem cell (MSC) origin and MSC proliferation and differentiation can have significant impacts on bone formation, it remains largely unknown whether regulation of MSC plays a role in the bone mass increase of Smurf1-deficient mice. In this study, we found that bone marrow mesenchymal progenitor cells from Smurf1-/- mice form significantly increased alkaline phosphatase-positive colonies, indicating roles of MSC proliferation and differentiation in bone mass accrual of Smurf1-/- mice. Interestingly, Smurf1-/- cells have an elevated protein level of AP-1 transcription factor JunB. Biochemical experiments demonstrate that Smurf1 interacts with JunB through PY motif and targets JunB protein for ubiquitination and proteasomal degradation. Indeed, Smurf1-deficient MSCs have higher proliferation rates, consistent with the facts that both mRNA and protein of cyclin D1 are increased in Smurf1-/- cells and JunB can induce cyclinD1 promoter. Moreover, JunB overexpression induces osteoblast differentiation shown by higher expression of osteoblast markers and JunB knocking-down not only decreases osteoblast differentiation but also restores the osteogenic potential to WT level in Smurf1-/- cells. In conclusion, our results suggest that Smurf1 negatively regulates MSC proliferation and differentiation by controlling JunB turnover through an ubiquitin-proteasome pathway. (c) 2010 American Society for Bone and Mineral Research.

Recombinant human myxovirus resistance protein A (MxA) was successfully expressed by an Escherichia coli expression system. After immunization and cell fusion, a mouse hybridoma (3C2) producing MAbs to MxA was established. Hybridoma 3C2 was further characterized using indirect ELISA, Western blot analysis, immunofluorescent staining, and immunoprecipitation. The ELISA results showed that the titer of 3C2 was between 1:6400 and 1:12800 in ascitic fluids. The isotype of the monoclonal antibody was tested to be IgG1kappa. 3C2 can also specifically recognize human MxA protein in various formats by Western blot analysis, immunofluorescent staining, and immunoprecipitation assay. We further demonstrated that 3C2 could be used to detected MxA expression induce by type I interferon in A549 cell line and human peripheral blood mononuclear cells by Western blot in a dose-dependent manner.

We have revisited the water-gas shift reaction catalyzed by iron pentacarbonyl at the DFT-B3LYP level. The reaction mechanism proposed by Rozanska and Vuilleumier (Inorg. Chem. 2008, 47, 8635-8640) has been followed and revised. The results show that transition states TS4/5 and TS5/2_a actually connect other intermediates rather than those suggested by Rozanska and Vuilleumier. Furthermore, the entire reaction has been proven to proceed with processes 1 --> 2 --> 3 --> 4 --> 6 --> 7 --> 2. It is the first time that species 6 and 7 are reported as intermediates for this reaction mechanism.

The major cause of cancer mortality is the metastatic spread of tumor cells that can occur via multiple routes, including the vascular system and the lymphatic system. In this study, we developed an IgG-like fusion protein molecule [vascular endothelial growth factor (VEGF) receptor 31-immunoglobulin (VEGFR31-Ig)] which could simultaneously bind the angiogenic growth factor VEGF-A and the lymphangiogenic growth factor VEGF-C. Importantly, VEGFR31-Ig exhibited VEGF-A-binding affinity similar to that of VEGFTrap, the most potent VEGF-A binder, and VEGF-C-binding affinity comparable with that of the soluble fusion protein VEGFR3-Ig (sVEGFR3). Pharmacokinetic analysis in mice showed that VEGFR31-Ig had improved pharmacokinetic properties compared with either VEGFTrap or sVEGFR3. In a highly metastatic human hepatocellular carcinoma (HCCLM3) model in severe combined immunodeficient mice, VEGFR31-Ig potently blocked both tumor angiogenesis and lymphangiogenesis, effectively inhibiting primary tumor growth and metastasis to lungs and lymph nodes. In contrast, VEGFTrap only suppressed primary tumor growth and metastasis to lungs by inhibiting tumor angiogenesis, whereas VEGFR3 was only effective in suppressing tumor metastasis to lymph nodes by blocking tumor lymphangiogenesis. Although a combination of VEGFTrap (25 mg/kg twice weekly) and sVEGFR3 (25 mg/kg twice weekly) can achieve the same therapeutic effect as VEGFR31-Ig (25 mg/kg twice weekly) in the HCCLM3 xenograft mouse model, developing two separate receptor-Ig fusion proteins for clinical use as combination therapy is impractical, mainly owing to regulatory hurdles and cost. Taken together, the VEGFR31-Ig fusion protein presented here has been suggested to have great potential for the treatment of metastatic cancer. Cancer Res; 70(6); 2495-503.

AIM OF THE STUDY: Cinobutacini, an aqueous extract from the skin and parotid venom glands of Bufo bufo gargarizans Cantor, is a traditional Chinese medicine widely used in clinical cancer therapy in China. The present study sought to investigate the possible signaling pathway implicated in cinobutacini-induced apoptosis in the hepatocellular carcinoma cell lines HepG(2) and Bel-7402. MATERIALS AND METHODS: The effects of cinobutacini on cell proliferation of HepG(2) and Bel-7402 cells were evaluated by 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide (MTT) assays. Cell apoptosis was detected by Hoechst 33258 staining and flow cytometry analysis. The mitochondrial membrane potential (Deltapsim) and caspase-9 and -3 activity were detected using MitoCapture reagent staining and colorimetric assays, respectively. The expression of apoptosis-related proteins and release of cytochrome c were assessed by Western blot analysis. RESULTS: Cinobutacini significantly inhibited cell proliferation of both cell lines in a dose- and time-dependent manner. Marked changes in apoptotic morphology and apoptosis rates were clearly observed after cinobutacini treatment. The protein expression of Bax increased whereas that of Bcl-2 decreased, leading to an increase in the Bax/Bcl-2 ratio. Subsequently, cinobutacini disrupted the mitochondrial membrane potential (Deltapsim) and resulted in the release of cytochrome c, activation of both caspase-9 and -3, and cleavage of poly (ADP-ribose) polymerase (PARP). CONCLUSION: The present study indicated that cinobutacini can induce apoptosis of HepG(2) and Bel-7402 cells through a mitochondria-mediated apoptosis pathway.

OBJECTIVE: To study clinical characteristics of ocular ischemic syndrome (OIS). METHODS: Retrospective study. Data, including sex, age, systemic diseases, best corrected visual acuity, anterior segment, fundus, fundus fluorescence angiography (FFA), transcranial Doppler (TCD) and digital subtraction angiography (DSA), were collected from 10 patients (12 eyes) with ocular ischemic syndrome from February 2008 to April 2009. RESULTS: The mean age was 59 years. There were six males and four females. All patients were combined with heart and/or cerebrovascular diseases. Best corrected visual acuity ranged from hand movement to 1.0. Neovascularization of the iris appeared in 5 eyes. Intraocular pressure over 21 mm Hg (1 mm Hg = 0.133 kPa) was present in 3 eyes. Fundus examination showed narrow retinal arteries, dilated but not tortuous veins, fleck-shaped hemorrhage of the retina, cotton-wool exudates, optic disc edema, optic disc neovascularization, and atypical macular cherry-red spots. TCD showed stenosis of internal carotid artery at different degrees. FFA was performed in 9 patients which showed artery front, prolonged arm-retinal circulation time, retinal circulation time and choroid circulation time. All patients accepted vasodilation and microcirculation improving treatments. Visual acuity was improved at different degrees. One patient (2 eyes) with iris neovascularization was treated with retinal photocoagulation and the neovascularization disappeared after the treatment. There were 3 patients (3 eyes) had neovascular glaucoma. One patient was treated by ciliary cryotherapy. One patient was treated by retinal photocoagulation and internal carotid artery stent implantation and one were treated by internal carotid artery stent implantation. Four patients were examined by DSA. Internal carotid artery stent implantation was performed in 3 patients, subclavian artery stent implantation was performed in one patient and internal carotid artery intima decollement was performed in one patient. CONCLUSIONS: Clinical manifestations of ocular ischemic syndrome patients are complicated which are depended on the different extents of ischemia. The management of ocular ischemic syndrome requires the collaboration of ophthalmologists, neurophysicians and neurosurgeons.

A numerical toolset to help in understanding physical processes in the electron cyclotron resonance charge breeder (ECRCB) and further to help optimization and design of current and future machines is presented. The toolset consists of three modules (Monte Carlo charge breeding code, generalized electron cyclotron resonance ion source modeling, and ion extraction), each modeling different processes occurring in the ECRCB from beam injection to extraction. The toolset provides qualitative study, such as parameter studies, and scaling of the operation, and physical understanding in the ECRCB. The methodology and a sample integrated modeling are presented.

A Monte Carlo charge breeding code (MCBC) is being developed by FAR-TECH, Inc. to model the capture and charge breeding of 1+ ion beam in an electron cyclotron resonance ion source (ECRIS) device. The ECRIS plasma is simulated using the generalized ECRIS model which has two choices of boundary settings, free boundary condition and Bohm condition. The charge state distribution of the extracted beam ions is calculated by solving the steady state ion continuity equations where the profiles of the captured ions are used as source terms. MCBC simulations of the charge breeding of Rb+ showed good agreement with recent charge breeding experiments at Argonne National Laboratory (ANL). MCBC correctly predicted the peak of highly charged ion state outputs under free boundary condition and similar charge state distribution width but a lower peak charge state under the Bohm condition. The comparisons between the simulation results and ANL experimental measurements are presented and discussed.

Although heating power and gas pressure are two of the two of primary experimental "knobs" available to users of electron cyclotron resonance ion sources, there is still no clear understanding of how they interact in order to provide optimal plasma conditions. FAR-TECH, Inc. has performed a series of simulations with its generalized electron cyclotron resonance ion source model in which the power and pressure were varied over a wide range. Analysis of the numerical data produces scaling laws that predict the plasma parameters as a function of the power and pressure. These scaling laws are in general agreement with experimental data.

Chronic exposure to morphine can induce drug addiction and neural injury, but the exact mechanism is not fully understood. Here we show that morphine induces autophagy in neuroblastoma SH-SY5Y cells and in the rat hippocampus. Pharmacological approach shows that this effect appears to be mediated by PTX-sensitive G protein-coupled receptors signaling cascade. Morphine increases Beclin 1 expression and reduces the interaction between Beclin 1 and Bcl-2, thus releasing Beclin 1 for its pro-autophagic activity. Bcl-2 overexpression inhibits morphine-induced autophagy, whereas knockdown of Beclin 1 or knockout of ATG5 prevents morphine-induced autophagy. In addition, chronic treatment with morphine induces cell death, which is increased by autophagy inhibition through Beclin 1 RNAi. Our data are the first to reveal that Beclin 1 and ATG5 play key roles in morphine-induced autophagy, which may contribute to morphine-induced neuronal injury.

OBJECTIVE: To develop and evaluate a simple tool, using data collected in a rural Chinese general practice, to identify those at high risk of Type 2 diabetes (T2DM) and prediabetes (PDM). STUDY DESIGN AND SETTING: A total of 2,261 rural Chinese participants without known diabetes were used to derive and validate the models of T2DM and T2DM plus PDM. Logistic regression and classification tree analysis were used to build models. RESULTS: The significant risk factors included in the logistic regression method were age, body mass index, waist/hip ratio (WHR), duration of hypertension, family history of diabetes, and history of hypertension for T2DM and T2DM plus PDM. In the classification tree analysis, WHR and duration of hypertension were the most important determining factors in the T2DM and T2DM plus PDM model. The sensitivity, specificity, positive predictive value, negative predictive value, and receiver operating characteristic area for detecting T2DM were 74.6%, 71.6%, 23.6%, 96.0%, and 0.731, respectively. For PDM plus T2DM, the results were 65.3%, 72.5%, 33.2%, 90.7%, and 0.689, respectively. CONCLUSION: The classification tree model is a simple and accurate tool to identify those at high risk of T2DM and PDM. Central obesity strongly associates with T2DM in rural Chinese.

OBJECTIVES: Persons with serious mental illnesses (SMI) have elevated rates of comorbid medical conditions, but may also face challenges in effectively managing those conditions. METHODS: The study team developed and pilot-tested the Health and Recovery Program (HARP), an adaptation of the Chronic Disease Self-Management Program (CDSMP) for mental health consumers. A manualized, six-session intervention, delivered by mental health peer leaders, helps participants become more effective managers of their chronic illnesses. A pilot trial randomized 80 consumers with one or more chronic medical illness to either the HARP program or usual care. RESULTS: At six month follow-up, participants in the HARP program had a significantly greater improvement in patient activation than those in usual care (7.7% relative improvement vs. 5.7% decline, p=0.03 for group *time interaction), and in rates of having one or more primary care visit (68.4% vs. 51.9% with one or more visit, p=0.046 for group *time interaction). Intervention advantages were observed for physical health related quality of life (HRQOL), physical activity, medication adherence, and, and though not statistically significant, had similar effect sizes as those seen for the CDSMP in general medical populations. Improvements in HRQOL were largest among medically and socially vulnerable subpopulations. CONCLUSIONS: This peer-led, medical self-management program was feasible and showed promise for improving a range of health outcomes among mental health consumers with chronic medical comorbidities. The HARP intervention may provide a vehicle for the mental health peer workforce to actively engage in efforts to reduce morbidity and mortality among mental health consumers.

OBJECTIVE: To evaluate the diagnosis and surgical treatment of obstructive azoospermia. METHODS: We analyzed the clinical data of 56 cases of obstructive azoospermia, 43 of them with ejaculatory duct obstruction (EDO), and the other 13 suspected of epididymal obstruction. The diagnostic methods included semen analyses, measurement of fructose and neutral alpha-glucosidase in the seminal plasma, transrectal ultrasonography (TRUS), and vasography when necessary. The 43 patients with EDO were treated by transurethral resection of the ejaculatory duct (TURED), and 11 of the 13 cases of suspected epididymal obstruction were confirmed by scrotal exploration and underwent either bilateral or unilateral vasoepididymostomy. The patients were followed up for 3 -51 months for postoperative semen quality and impregnation. RESULTS: Of the 43 azoospermia patients with EDO treated by TURED, 36 (83.7%) showed improved semen parameters and 11 (25.6%) achieved pregnancies. Among the 11 cases of azoospermia with confirmed epididymal obstruction treated by vasoepididymostomy, 6 (54.5%) had sperm in the semen assay and 3 (27.3%) achieved pregnancies. CONCLUSION: Semen analyses, measurement of fructose and neutral alpha-glucosidase in the seminal plasma, TRUS and vasography are important diagnostic methods for obstructive azoospermia. TURED is effective for azoospermia with EDO, while vasoepididymostomy is preferable for the treatment of azoospermia with epididymal obstruction.

A direct method to determine the pitch and chirality of complicated hierarchical helical mesostructures is presented by using the state-of-the-art electron tomography technique.

Anatase TiO(2) single crystals with exposed {001} and {110} facets have been successfully synthesized using a modified hydrothermal technique in the presence of hydrogen peroxide and hydrofluoric acid solution; these single crystals exhibited enhanced photocatalytic activities for degradation of Methylene Blue dye under ultraviolet light irradiation.

Humans are superorganisms with two genomes that dictate phenotype, the genetically inherited human genome (25,000 genes) and the environmentally acquired human microbiome (over 1 million genes). The two genomes must work in harmonious integration as a hologenome to maintain health. Nutrition plays a crucial role in directly modulating our microbiomes and health phenotypes. Poorly balanced diets can turn the gut microbiome from a partner for health to a "pathogen" in chronic diseases, e.g. accumulating evidence supports the new hypothesis that obesity and related metabolic diseases develop because of low-grade, systemic and chronic inflammation induced by diet-disrupted gut microbiota. Due to the tight integration of gut microbiota into human global metabolism, molecular profiling of urine metabolites can provide a new window for reflecting physiological functions of gut microbiomes. Changes of gut microbiota and urine metabolites can thus be employed as new systems approaches for quantitative assessment and monitoring of health at the whole-body level with the advantage of measuring human health based on the results of interactions between the two genomes and the environment rather than just host genomic information. Large scale population-based studies in conjunction with these whole-body level systems methods will generate pre-disease biomarkers with predictive power, thus making preventive health management of populations with rapidly changing disease spectrums possible through re-engineering of the imbalanced gut microbiomes with specially designed foods/diets.