Discovery of novel iminosydnone compounds with insecticidal activities based on the binding mode of triflumezopyrim.

Triflumezopyrim (TFM) is a new mesoionic insecticide developed by DuPont. Like other neonicotinoid insecticides, it binds to the orthosteric site of the nicotinic acetylcholine receptor (nAChR), but the binding mode has not been reported. Nicotinic acetylcholine binding proteins (nAChBPs) are ideal alternative structure for nAChRs. In this study, molecular docking, molecular dynamics (MD) simulations, binding free energy calculation, and per-residue binding free energy decomposition were used to study the binding modes of TFM and other 12 mesoionic insecticides. By comparing the binding free energy and the insecticidal activity, it was found that the sub-pocket around the benzyl group of the mesoionic insecticide is the key area for maintaining its activity, which is composed of A: Val116, A: Met124, A: Ile126, B: Trp155 and B: Val156. In order to verify the druggability of the sub-pocket, a series of iminosydnone compounds were designed and synthesized based on the structure of the sub-pocket. The lethality rate of compound 1 against Mythimna separata were 100% at 500 mg/L. Our research provides a basis for designing new mesoionic insecticides based on structure.

Linking servant leadership to followers' thriving at work: self-determination theory perspective.

Previous studies have confirmed that servant leadership has a positive impact on thriving at work, however, the psychological mechanism in this process has not been fully understood. Based on Self-Determination Theory, this study examines the mediating effect of basic psychological needs and the moderating effect of power distance on the relationship between servant leadership and followers' thriving at work. The results from the between-subject experimental design (Study 1) indicate that servant leadership can satisfy followers' three basic psychological needs. And the results from a questionnaire survey of 455 civil servants at two-time points (Study 2) indicate: (1) Servant leadership has a significantly positive impact on followers' thriving at work; (2) All three basic psychological needs satisfaction serve as a mediator in the relationship between servant leadership and followers' thriving at work; (3) Power distance negatively moderates the relationship between servant leadership and the satisfaction of three basic psychological needs, meaning that the lower on the power distance, the stronger the positive relationship between servant leadership and the satisfaction of three basic psychological needs; (4) Power distance negatively moderates the mediating effects of competence needs satisfaction and relatedness needs satisfaction in the relationship between servant leadership and followers' thriving at work, indicating that the lower on the power distance, the stronger the mediating effects. Our findings highlight the important role of servant leadership in fostering followers' thriving at work and explore the critical role of basic psychological needs satisfaction. This provides empirical evidence to further refine theories regarding thriving at work, and suggests that in order to promote employee thriving, it is important to guide leaders to reevaluating and repositioning their roles.

Calcium antagonists for acute ischemic stroke.

BACKGROUND: The sudden loss of blood supply in ischemic stroke is associated with the increase of calcium ions within neurons. Inhibiting this increase could protect neurons and hence might reduce neurological impairment, disability and handicap after stroke. OBJECTIVES: To determine whether calcium antagonists reduce the risk of death or dependency after acute ischemic stroke. To investigate the influence of different drugs, dosages, routes of administration, time intervals after stroke and trial design on the risk of a primary outcome. SEARCH METHODS: We searched the Cochrane Stroke Group Trials Register (January 2012), MEDLINE (1950 to December 2011), EMBASE (1980 to December 2011), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library, 2011 issue 4) and four Chinese databases (December 2011): Chinese Biological Medicine Database (CBM-disc), China National Knowledge Infrastructure (CNKI), Chinese scientific periodical database of VIP information and Wanfang Data. We also contacted trialists and researchers. SELECTION CRITERIA: All truly randomized trials comparing a calcium antagonist with control in patients with acute ischemic stroke. DATA COLLECTION AND ANALYSIS: Two authors assessed all trials and extracted the data. We used death or dependency at the end of long-term follow-up (at least three months) in activities of daily living as the primary outcome. Analyses were, if possible, intention-to-treat. MAIN RESULTS: We included 34 trials including 7731 patients. There was no effect of calcium antagonists on the primary outcome (risk ratio (RR) 1.05; 95% confidence interval (CI) 0.98 to 1.13), or on death at the end of follow-up (RR 1.07, 95% CI 0.98 to 1.17). Comparisons of different doses of nimodipine suggested that the highest doses were associated with poorer outcome. AUTHORS' CONCLUSIONS: No evidence is available using calcium antagonists in patients with acute ischemic stroke is effective.

Release of copper and indomethacin from intrauterine devices immersed in simulated uterine fluid.

OBJECTIVES: To study the release of cupric ions and indomethacin from copper-bearing intrauterine devices (Cu-IUDs) and to assess the influence of their surface area of copper and the pH of the test medium on the release of cupric ions. METHODS: Cu-IUDs were incubated in simulated uterine fluid (SUF). The in vitro release of cupric ions and indomethacin was determined by means of a flame atomic absorption spectrometer and a UV 752 spectrophotometer, respectively, over a period of 220 days. The effect of indomethacin on the pH of the SUF was monitored during the cupric ions release study. RESULTS: The Cu-IUDs released cupric ions in vitro according to a biphasic pattern. After an initial burst release, the rate slowed down and then became steady. The release of indomethacin from two medicated Cu-IUDs was biphasic and, in both cases, in accordance with the in vitro dissolution model of the drug. The surface area of copper of Cu-IUDs and the pH change induced by indomethacin incorporated in their frame affected the release rate of copper (p < 0.05). CONCLUSION: The surface area of the copper and the pH of the test medium modulate the in vitro release of cupric ions from Cu-IUDs.

Rapid Detection of Candida albicans by Polymerase Spiral Reaction Assay in Clinical Blood Samples.

Candida albicans is the most common human yeast pathogen which causes mucosal infections and invasive fungal diseases. Early detection of this pathogen is needed to guide preventative and therapeutic treatment. The aim of this study was to establish a polymerase spiral reaction (PSR) assay that rapidly and accurately detects C. albicans and to assess the clinical applicability of PSR-based diagnostic testing. Internal transcribed spacer 2 (ITS2), a region between 5.8S and 28S fungal ribosomal DNA, was used as the target sequence. Four primers were designed for amplification of ITS2 with the PSR method, which was evaluated using real time turbidity monitoring and visual detection using a pH indicator. Fourteen non-C. albicans yeast strains were negative for detection, which indicated the specificity of PSR assay was 100%. A 10-fold serial dilution of C. albicans genomic DNA was subjected to PSR and conventional polimerase chain reaction (PCR) to compare their sensitivities. The detection limit of PSR was 6.9 pg/µl within 1 h, 10-fold higher than that of PCR (69.0 pg/µl). Blood samples (n = 122) were collected from intensive care unit and hematological patients with proven or suspected C. albicans infection at two hospitals in Beijing, China. Both PSR assay and the culture method were used to analyze the samples. Of the 122 clinical samples, 34 were identified as positive by PSR. The result was consistent with those obtained by the culture method. In conclusion, a novel and effective C. albicans detection assay was developed that has a great potential for clinical screening and point-of-care testing.

Melittin-induced membrane permeability: a nonosmotic mechanism of cell death.

Derived from honeybees, melittin is a 26-amino acid, alpha-helical, membrane-attack protein that efficiently kills mammalian cells. To investigate the contribution of colloid-osmotic effects to the mechanism of cell death, we studied the effect of melittin on lymphocyte membrane permeability and cell volumes. Melittin concentrations of 0.5 to 2.0 microM induced release of membrane permeability markers without total disruption of the cell membrane. At these melittin concentrations, electrical-impedance cytometry demonstrated melittin-induced changes in red blood cell volumes (P<0.01), but no change in lymphocyte cell volumes (P>0.05). Streaming video microscopy, obtaining images of melittin-treated lymphocytes at 80-ms intervals, demonstrated a loss of optical density (P<0.001) suggesting a flattening of the cell but no significant increase in cell perimeter (P>0.05). Real-time multiparameter flow cytometry of melittin-treated lymphocytes confirmed simultaneous loss of the cytoplasmic marker, calcein, and uptake of the DNA dye, ethidium homodimer, but demonstrated no increase in forward light scatter. Transmission-electron microscopy of melittin-treated lymphocytes showed normal cell volumes but discontinuities in the cell membrane suggesting direct membrane toxicity. We conclude that melittin causes lymphocyte death by a "leaky patch" mechanism that is independent of colloid-osmotic effects.

Antithrombotic therapy for ischaemic stroke patients with AF/RHD in West China daily practice.

BACKGROUND AND OBJECTIVE: Although, current evidence-based guidelines advocate anticoagulation as a strong recommendation in IS patients with AF/RHD, the underuse of anticoagulation in IS patients with AF/RHD has been found in clinical practice worldwide. Nevertheless, there was little information about implementation of antithrombotic therapy to prevent stroke for patients with AF and/or RHD in daily practice in West China. Our study determined to clarify the patterns, adherence and comparative effect of antithrombotic treatment during 1-year follow-up in IS patients with AF and/or RHD. METHOD: Consecutive patients with acute IS and AF/RHD admitted to Department of Neurology, West China Hospital from November 2010 to October 2011 were included in the study. RESULTS: 155 consecutive patients were analysed in this study. One hundred thirteen patients have been diagnosed as AF and/or RHD before admission. Of these, 49 (43.4%) patients were receiving antithrombotic therapy before the time of admission, including nine (8.0%) patients receiving warfarin. At 12  months after stroke onset, 109 (81.3%) patients were on antithrombotics, and 46 (34.3%) patients were on warfarin alone. The persistence rate of warfarin use at 1 year was 77.8%. Moreover, there were 80 (81.6%) patients never starting to use warfarin. Compared with no antithrombotic therapy, anti-platelets and warfarin reduced death risk significantly during 1-year after stroke onset (P = 0.005). CONCLUSION: Our study suggests that overall real-world use of warfarin in IS patients with AF and/or RHD is low before and after admission in West China. Implementation study on this respect should be conducted in this area to improve the daily practice.

Structural mapping of immunoglobulin subclasses using multiplexed bead flow cytometry.

Monoclonal hybridomas secrete immunoglobulin molecules with a single specificity and distinct class/subclass structure. The determination of immunoglobulin structure can be used to facilitate hybridoma colony management and predict monoclonality. In this report, we used multiplexed bead flow cytometry to define hybridoma class/subclass. The assay was sufficiently sensitive to detect 50 ng/mL of antibody. The multiplexed bead assay efficiently defined traditional class/subclass determinants as well as more subtle patterns of crossreactivity. Further, the assay was combined with Poisson statistics to provide a numerical estimate of hybridoma monoclonality. The sensitivity and flexibility of this approach suggests the utility of multiplexed bead flow cytometry in the early management of immunoglobulin-secreting hybridomas.

Epstein-Barr virus with the latent infection nuclear antigen 3B completely deleted is still competent for B-cell growth transformation in vitro.

The Epstein-Barr virus (EBV) nuclear antigen 3B (EBNA-3B) is considered nonessential for EBV-mediated B-cell growth transformation in vitro based on three virus isolates with EBNA-3B mutations. Two of these isolates could potentially express truncated EBNA-3B products, and, similarly, we now show that the third isolate, IB4, has a point mutation and in-frame deletion of 263 amino acids. In order to test whether a virus with EBNA-3B completely deleted can immortalize B-cell growth, we first cloned the EBV genome as a bacterial artificial chromosome (BAC) and showed that the BAC-derived virus was B-cell immortalization competent. Deletion of the entire EBNA-3B open reading frame from the EBV BAC had no adverse impact on growth of EBV-immortalized B cells, providing formal proof that EBNA-3B is not essential for EBV-mediated B-cell growth transformation in vitro.

Vessel painting of the microcirculation using fluorescent lipophilic tracers.

Flexible approaches to defining microvessel morphometry are useful in the study of both acute and chronic structural changes of the microcirculation. In this report, we examined the utility of the intravascular infusion of lipophilic carbocyanine tracers in the structural assessment of the retina, skin, lung, and colon microcirculation. The microvessel labeling technique, here termed fluorescent vessel painting, involved the intravascular injection of sulfonated lipophilic carbocyanine tracers. The utility of vessel painting in morphometry was assessed using morphometric comparisons with corrosion casting and 2-dimensional and 3-dimensional scanning electron microscopy. The comparisons demonstrated that fluorescent vessel painting modestly overestimated the interbranch angles, interbranch distances, and vessel diameters of the 2D mucosal plexus of the colon. These differences were narrowed with the application of confocal microscopy. The advantages of fluorescence vessel painting included (1) the filling of all tissues including the relatively high resistance microvessels of the mouse skin, (2) the ability to use tissue counterstains such as DAPI, and (3) the prolonged stability of the lipophilic tracer after aldehyde fixation. These studies suggest the utility of fluorescent vessel painting as a complementary technique to corrosion casting in the morphometric study of the microcirculation.

Novel nonnucleoside inhibitor of hepatitis C virus RNA-dependent RNA polymerase.

A novel nonnucleoside inhibitor of hepatitis C virus (HCV) RNA-dependent RNA polymerase (RdRp), [(1R)-5-cyano-8-methyl-1-propyl-1,3,4,9-tetrahydropyano[3,4-b]indol-1-yl] acetic acid (HCV-371), was discovered through high-throughput screening followed by chemical optimization. HCV-371 displayed broad inhibitory activities against the NS5B RdRp enzyme, with 50% inhibitory concentrations ranging from 0.3 to 1.8 microM for 90% of the isolates derived from HCV genotypes 1a, 1b, and 3a. HCV-371 showed no inhibitory activity against a panel of human polymerases, including mitochondrial DNA polymerase gamma, and other unrelated viral polymerases, demonstrating its specificity for the HCV polymerase. A single administration of HCV-371 to cells containing the HCV subgenomic replicon for 3 days resulted in a dose-dependent reduction of the steady-state levels of viral RNA and protein. Multiple treatments with HCV-371 for 16 days led to a >3-log10 reduction in the HCV RNA level. In comparison, multiple treatments with a similar inhibitory dose of alpha interferon resulted in a 2-log10 reduction of the viral RNA level. In addition, treatment of cells with a combination of HCV-371 and pegylated alpha interferon resulted in an additive antiviral activity. Within the effective antiviral concentrations of HCV-371, there was no effect on cell viability and metabolism. The intracellular antiviral specificity of HCV-371 was demonstrated by its lack of activity in cells infected with several DNA or RNA viruses. Fluorescence binding studies show that HCV-371 binds the NS5B with an apparent dissociation constant of 150 nM, leading to high selectivity and lack of cytotoxicity in the antiviral assays.