No abstract

Many G protein-coupled receptors possess carboxyl-terminal motifs ideal for interaction with PDZ scaffold proteins, which can control receptor trafficking and signaling in a cell-specific manner. To gain a panoramic view of ␤ 1 -adrenergic receptor (␤ 1 AR) interactions with PDZ scaffolds, the ␤ 1 AR carboxyl terminus was screened against a newly developed proteomic array of PDZ domains. These screens confirmed ␤ 1 AR associations with several previously identified PDZ partners, such as PSD-95, MAGI-2, GIPC, and CAL. Moreover, two novel ␤ 1 AR-interacting proteins, SAP97 and MAGI-3, were also identified. The ␤ 1 AR carboxyl terminus was found to bind specifically to the first PDZ domain of MAGI-3, with the last four amino acids (E-S-K-V) of ␤ 1 AR being the key determinants of the interaction. Full-length ␤ 1 AR robustly associated with full-length MAGI-3 in cells, and this association was abolished by mutation of the ␤ 1 AR terminal valine residue to alanine (V477A), as determined by co-immunoprecipitation experiments and immunofluorescence co-localization studies. MAGI-3 co-expression with ␤ 1 AR profoundly impaired ␤ 1 AR-mediated ERK1/2 activation but had no apparent effect on ␤ 1 AR-mediated cyclic AMP generation or agonist-promoted ␤ 1 AR internalization. These findings revealed that the interaction of MAGI-3 with ␤ 1 AR can selectively regulate specific aspects of receptor signaling. Moreover, the screens of the PDZ domain proteomic array provide a comprehensive view of ␤ 1 AR interactions with PDZ scaffolds, thereby shedding light on the molecular mechanisms by which ␤ 1 AR signaling and trafficking can be regulated in a cell-specific manner.Cellular responses to a given hormone or neurotransmitter can vary markedly between different types of cells. In many cases, such cellular differences are because of differential expression of receptor subtypes. However, even cells expressing exactly the same receptor subtypes can exhibit distinct responses to a given ligand, because receptors often behave quite differently in distinct cellular environments. The trafficking and signaling properties of G protein-coupled receptors (GPCRs), 3 which comprise the largest family of cell surface receptors, are known to be especially influenced by cellular context. The activity of a GPCR depends not only on the complement of downstream effectors expressed in a given cell but also on the set of expressed G proteins, kinases, and scaffold proteins that directly interact with the receptor to shape its signaling capability.PDZ scaffolds comprise a key class of GPCR-interacting proteins that can strongly influence receptor trafficking and signaling. The term PDZ is derived from the names of the first three proteins in which these domains were first identified: the post-synaptic density protein PSD-95, the Drosophila protein Discs-large, and the tight junction protein ZO-1. PDZ domains are ϳ90 amino acids in length and bind to specific carboxyl-terminal motifs on their target proteins. There are three general classes of PDZ domain...

Neuronal growth cones are motile structures located at the end of axons that translate extracellular guidance information into directional movements. Despite the important role of growth cones in neuronal development and regeneration, relatively little is known about the topography and mechanical properties of distinct subcellular growth cone regions under live conditions. In this study, we used the AFM to study the P domain, T zone, and C domain of live Aplysia growth cones. The average height of these regions was calculated from contact mode AFM images to be 183 +/- 33, 690 +/- 274, and 1322 +/- 164 nm, respectively. These findings are consistent with data derived from dynamic mode images of live and contact mode images of fixed growth cones. Nano-indentation measurements indicate that the elastic moduli of the C domain and T zone ruffling region ranged between 3-7 and 7-23 kPa, respectively. The range of the measured elastic modulus of the P domain was 10-40 kPa. High resolution images of the P domain suggest its relatively high elastic modulus results from a dense meshwork of actin filaments in lamellipodia and from actin bundles in the filopodia. The increased mechanical stiffness of the P and T domains is likely important to support and transduce tension that develops during growth cone steering.

Label-free microfluidic paper-based electrochemical aptasensor for ultrasensitive and simultaneous multiplexed detection of cancer biomarkers. Biosensors and Bioelectronics, 136, pp. 84-90. There may be differences between this version and the published version. You are advised to consult the publisher's version if you wish to cite from it.

No abstract

IMPORTANCE A randomized clinical trial is needed to evaluate what is the best photodynamic therapy (PDT) protocol to use for acute central serous chorioretinopathy.OBJECTIVE To compare the efficacy and safety of a 50% dose of verteporfin (a method of PDT) with the efficacy and safety of a 30% dose for acute central serous chorioretinopathy. DESIGN, SETTING, AND PARTICIPANTSA multicenter, noninferiority, double-masked, randomized, controlled, clinical trial in which 131 patients (131 eyes) with acute central serous chorioretinopathy for less than 6 months were recruited with a follow-up of 12 months from university-based ophthalmology practices.INTERVENTIONS Patients were randomly assigned to either a 50% dose of verteporfin (the 50%-dose PDT group) or a 30% dose (the 30%-dose PDT group). MAIN OUTCOMES AND MEASURESThe 2 primary outcome measures were the proportion of eyes with complete absorption of subretinal fluid and the proportion of eyes with complete disappearance of fluorescein leakage at 6 and 12 months. The secondary outcome measures included the subretinal fluid recurrent rate, the fluorescein leakage recurrent rate at 12 months, the mean best-corrected visual acuity, the retinal thickness of the foveal center, and the maximum retinal thickness at each scheduled visit. RESULTSThe noninferiority of the 30%-dose PDT compared with the 50%-dose PDT for the primary outcomes was not demonstrated. The optical coherence tomography-based improvement rate in the 30%-dose PDT group was less than that in the 50%-dose PDT group both at 6 months (73.8% vs 92.9%; α = 0.0125, P = .006) and at 12 months (75.4% vs 94.6%; α = 0.0125, P = .004). The fluorescein angiography-based improvement rate in the 30%-dose PDT group was less than that in the 50%-dose PDT group both at 6 months (68.9% vs 91.1%; α = 0.0125, P = .003) and at 12 months (68.9% vs 92.9%; α = 0.0125, P = .001). The subretinal fluid recurrence rate in the 30%-dose PDT group was greater than that in the 50%-dose PDT group (24.0% vs 5.7% at 12 months; P = .010, determined by use of the log-rank test). The fluorescein leakage recurrent rate in the 30%-dose PDT group was significantly higher than that in the 50%-dose PDT group (16.7% vs 3.8% at 12 months; P = .03, determined by use of the log-rank test). No ocular adverse event was encountered in the study. CONCLUSIONS AND RELEVANCEA 50% dose of verteporfin may be more effective at resolving subretinal fluid and fluorescein leakage, and with better visual outcomes, than a 30% dose for acute central serous chorioretinopathy. TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT01574430

Angiogenesis has an important role in tumour cell growth and metastasis. Anisomycin has been shown to inhibit tumour cell growth. However, whether anisomycin can inhibit angiogenesis of tumours has not been reported. The present study demonstrated that there was a positive correlation between tumour angiogenesis and the number of CD44 + /CD133 + serous human ovarian cancer stem cells (HuOCSCs). Subsequently, it was confirmed that anisomycin significantly inhibited the proliferation, invasion, tumorigenic ability and tumour angiogenesis of HuOCSCs. Gene expression profiling by cDNA microarrays revealed that the expression levels of vascular endothelial cell markers, platelet-derived growth factors, Notch pathway components and 27 tumour angiogenesis-related genes were significantly decreased in the anisomycin-treated group compared with the control group. Further experiments demonstrated that the expression levels of endogenous long non-coding RNA (lncRNA) maternally expressed 3 (Meg3) were significantly decreased in anisomycin-treated HuOCSCs, whereas the expression levels of microRNA (miR)-421 were significantly increased. The results of luciferase reporter assays indicated that, when miR-421 was overexpressed in cells, the luciferase activities of wild-type platelet derived growth factor receptor α (PDGFRA) 3' untranslated region and Meg3 reporter plasmids were significantly decreased. Overexpression of miR-421 in HuOCSCs significantly enhanced the anisomycin-mediated inhibition of HuOCSC proliferation. Taken together, the present results demonstrated that anisomycin inhibited the activation downstream of the Notch1 pathway by attenuating the molecular sponge effect of the lncRNA-Meg3/miR-421/PDGFRA axis, ultimately inhibiting angiogenesis, proliferation and invasion in ovarian cancer cells.

We compare multifocal intraocular lenses (MFIOLs) to monofocal IOLs for visual acuity (VA), contrast sensitivity, and adverse events using data from 21 randomized controlled trials with 2951 subjects. There was no statistical difference between uncorrected distance VA and corrected distance VA. Compared with monofocal IOLs, MFIOLs showed a better performance on uncorrected intermediate VA measured at 60 cm and uncorrected near VA; the mean differences were-0.06 (95% confidence interval [