Synaptic Targeting and Function of SAPAPs Mediated by Phosphorylation-Dependent Binding to PSD-95 MAGUKs

The PSD-95/SAPAP/Shank complex functions as the major scaffold in orchestrating the formation and plasticity of the post-synaptic densities (PSDs). We previously demonstrated that the exquisitely specific SAPAP/Shank interaction is critical for Shank synaptic targeting and Shank-mediated synaptogenesis. Here, we show that the PSD-95/SAPAP interaction, SAPAP synaptic targeting, and SAPAP-mediated synaptogenesis require phosphorylation of the N-terminal repeat sequences of SAPAPs. The atomic structure of the PSD-95 guanylate kinase (GK) in complex with a phosphor-SAPAP repeat peptide, together with biochemical studies, reveals the molecular mechanism underlying the phosphorylation-dependent PSD-95/SAPAP interaction, and it also provides an explanation of a PSD-95 mutation found in patients with intellectual disabilities. Guided by the structural data, we developed potent non-phosphorylated GK inhibitory peptides capable of blocking the PSD-95/SAPAP interaction and interfering with PSD-95/SAPAP-mediated synaptic maturation and strength. These peptides are genetically encodable for investigating the functions of the PSD-95/SAPAP interaction in vivo. Using structural biology, cell biology, and electrophysiology approaches, Zhu et al. demonstrate that phosphorylation of the N-terminal repeating sequences of SAPAPs is required for the SAPAP/PSD-95 complex formation and SAPAP's synaptic targeting and maturation functions. They also developed a potent non-phosphorylated PSD-95 GK inhibitory peptide that can effectively disrupt the SAPAP/PSD-95 complex formation and thus inhibit excitatory synaptic activities. Keywords: GK domain; PSD-95; SAPAP; MAGUK; postsynaptic density; synaptic scaffold proteins; synaptogenesis; synaptic plasticit

Binocular Goggle Augmented Imaging and Navigation System provides real-time fluorescence image guidance for tumor resection and sentinel lymph node mapping

The inability to identify microscopic tumors and assess surgical margins in real-time during oncologic surgery leads to incomplete tumor removal, increases the chances of tumor recurrence, and necessitates costly repeat surgery. To overcome these challenges, we have developed a wearable goggle augmented imaging and navigation system (GAINS) that can provide accurate intraoperative visualization of tumors and sentinel lymph nodes in real-time without disrupting normal surgical workflow. GAINS projects both near-infrared fluorescence from tumors and the natural color images of tissue onto a head-mounted display without latency. Aided by tumor-targeted contrast agents, the system detected tumors in subcutaneous and metastatic mouse models with high accuracy (sensitivity = 100%, specificity = 98% ± 5% standard deviation). Human pilot studies in breast cancer and melanoma patients using a near-infrared dye show that the GAINS detected sentinel lymph nodes with 100% sensitivity. Clinical use of the GAINS to guide tumor resection and sentinel lymph node mapping promises to improve surgical outcomes, reduce rates of repeat surgery, and improve the accuracy of cancer staging

Pins is not required for spindle orientation in the Drosophila wing disc.

In animal cells, mitotic spindles are oriented by the dynein/dynactin motor complex, which exerts a pulling force on astral microtubules. Dynein/dynactin localization depends on Mud/NUMA, which is typically recruited to the cortex by Pins/LGN. In Drosophila neuroblasts, the Inscuteable/Baz/Par-6/aPKC complex recruits Pins apically to induce vertical spindle orientation, whereas in epithelial cells Dlg recruits Pins laterally to orient the spindle horizontally. Here we investigate division orientation in the Drosophila imaginal wing disc epithelium. Live imaging reveals that spindle angles vary widely during prometaphase and metaphase, and therefore do not reliably predict division orientation. This finding prompted us to re-examine mutants that have been reported to disrupt division orientation in this tissue. Loss of Mud misorients divisions, but Inscuteable expression and aPKC, dlg and pins mutants have no effect. Furthermore, Mud localizes to the apical-lateral cortex of the wing epithelium independently of both Pins and cell cycle stage. Thus, Pins is not required in the wing disc because there are parallel mechanisms for Mud localization and hence spindle orientation, making it a more robust system than in other epithelia.This work was supported by a Wellcome Trust Principal Fellowship to DStJ [080007] and by core support from the Wellcome Trust [092096] and Cancer Research UK [A14492]. DTB was supported by a Marie Curie Fellowship and the Wellcome Trust. HEL was supported by a Herchel Smith Studentship.This is the author accepted manuscript. The final version is available from The Company of Biologists via http://dx.doi.org/10.1242/dev.13547

A-769662 Protects Osteoblasts from Hydrogen Dioxide-Induced Apoptosis through Activating of AMP-Activated Protein Kinase (AMPK)

Here we report that 5'-monophosphate (AMP)-activated protein kinase (AMPK) agonist A-769662 inhibited hydrogen peroxide (H2O2)-induced viability loss and apoptosis of human and mouse osteoblast cells. H2O2-induced moderate AMPK activation in osteoblast cells, which was enhanced by A-769662. Inactivation of AMPK by its inhibitor compound C, or by target shRNA-mediated silencing and kinase dead (KD) mutation exacerbated H2O2-induced cytotoxicity in osteoblast cells. A-769662-mediated protective effect against H2O2 was also blocked by AMPK inhibition or depletion. A-769662 inhibited reactive oxygen species (ROS) accumulation by H2O2 in osteoblast cells. Meanwhile, H2O2-induced ATP depletion was inhibited by A-769662, but was aggravated by compound C. Further, H2O2 induced AMPK-dependent and pro-survival autophagy in cultured osteoblast cells, which was enhanced by A-769662. Our results suggested that activation of AMPK by H2O2 is anti-apoptosis and pro-survival in osteoblast cells, probably due to its anti-oxidant, pro-autophagy and ATP preservation abilities, and A-769662-mediated cell-protective effect in osteoblast cells requires AMPK activation. Our study suggests that A-769662 might be further investigated as a novel anti-osteonecrosis agent

Catadioptric freeform optical system design for LED off-axis road illumination applications

The aim of this paper is to develop a new composite structure of catadioptric optical system containing both freeform refractive surface and freeform total internal reflective (TIR) surface for LED road illumination applications. The role of freeform refractive part is to generate the shifted general rectangular illumination pattern to optimally match the shape of the road surface. The application of TIR mechanism is aimed to control the stray light in the sidewalk direction of the road luminaire and maximize the efficient energy efficiency. In this paper, we use the "double pole" ray mapping technique to design the refractive optical surface and the theta-phi coordinate ray mapping technique to derive the freeform TIR surface. The simulation shows that the novel catadioptric design has relatively high collection efficiency, thus high average illuminance level inside the effective illumination area. This lens also has good control of stray light on the backside of the road luminaire. (c) 2017 Optical Society of America under the terms of the OSA Open Access Publishing AgreementWuhan Science and Technology Bureau [2017010201010110]; Huazhong University of Science and Technology [2017KFYXJJ026]Open access journal.This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Structural analyses of key features in the KANK1·KIF21A complex yield mechanistic insights into the cross-talk between microtubules and the cell cortex

The cross-talk between dynamic microtubules and the cell cortex plays important roles in cell division, polarity, and migration. A critical adaptor that links the plus ends of microtubules with the cell cortex is the KANK N-terminal motif and ankyrin repeat domains 1 (KANK1)/kinesin family member 21A (KIF21A) complex. Genetic defects in these two proteins are associated with various cancers and developmental diseases, such as congenital fibrosis of the extraocular muscles type 1. However, the molecular mechanism governing the KANK1/KIF21A interaction and the role of the conserved ankyrin (ANK) repeats in this interaction are still unclear. In this study, we present the crystal structure of the KANK1 center dot KIF21A complex at 2.1 angstrom resolution. The structure, together with biochemical studies, revealed that a five-helix-bundle-capping domain immediately preceding the ANK repeats of KANK1 forms a structural and functional supramodule with its ANK repeats in binding to an evolutionarily conserved peptide located in the middle of KIF21A. We also show that several missense mutations present in cancer patients are located at the interface of the KANK1 center dot KIF21A complex and destabilize its formation. In conclusion, our study elucidates the molecular basis underlying the KANK1/KIF21A interaction and also provides possible mechanistic explanations for the diseases caused by mutations in KANK1 and KIF21A.National Natural Science Foundation of China [31470733, U1532121]; Shanghai Municipal Science and Technology Commission, China ("Yang-Fan program") Grant [14YF1406700]; Strategic Priority Research Program of the Chinese Academy of Sciences [XDB08030104]12 month embargo; published online: 20 November 2017This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Discrimination of Minced Mutton Adulteration Based on Sized-Adaptive Online NIRS Information and 2D Conventional Neural Network

Single-probe near-infrared spectroscopy (NIRS) usually uses different spectral information for modelling, but there are few reports about its influence on model performance. Based on sized-adaptive online NIRS information and the 2D conventional neural network (CNN), minced samples of pure mutton, pork, duck, and adulterated mutton with pork/duck were classified in this study. The influence of spectral information, convolution kernel sizes, and classifiers on model performance was separately explored. The results showed that spectral information had a great influence on model accuracy, of which the maximum difference could reach up to 12.06% for the same validation set. The convolution kernel sizes and classifiers had little effect on model accuracy but had significant influence on classification speed. For all datasets, the accuracy of the CNN model with mean spectral information per direction, extreme learning machine (ELM) classifier, and 7 × 7 convolution kernel was higher than 99.56%. Considering the rapidity and practicality, this study provides a fast and accurate method for online classification of adulterated mutton

Quantitative Detection of Chromium Pollution in Biochar Based on Matrix Effect Classification Regression Model

Returning biochar to farmland has become one of the nationally promoted technologies for soil remediation and improvement in China. Rapid detection of heavy metals in biochar derived from varied materials can provide a guarantee for contaminated soil, avoiding secondary pollution. This work aims first to apply laser-induced breakdown spectroscopy (LIBS) for the quantitative detection of Cr in biochar. Learning from the principles of traditional matrix effect correction methods, calibration samples were divided into 1–3 classifications by an unsupervised hierarchical clustering method based on the main elemental LIBS data in biochar. The prediction samples were then divided into diverse classifications of calibration samples by a supervised K-nearest neighbor (KNN) algorithm. By comparing the effects of multiple partial least squares regression (PLSR) models, the results show that larger numbered classifications have a lower averaged relative standard deviations of cross-validation (ARSDCV) value, signifying a better calibration performance. Therefore, the 3 classification regression model was employed in this study, which had a better prediction performance with a lower averaged relative standard deviations of prediction (ARSDP) value of 8.13%, in comparison with our previous research and related literature results. The LIBS technology combined with matrix effect classification regression model can weaken the influence of the complex matrix effect of biochar and achieve accurate quantification of contaminated metal Cr in biochar