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https://www.readbyqxmd.com/read/28922398/role-of-the-p2-residue-of-human-alpha-1-antitrypsin-in-determining-target-protease-specificity
#1
Hye-Shin Chung, Ji-Sun Kim, Sang Mee Lee, Soon Jae Park
Alpha 1-antitrypsin (A1AT) is a serine protease inhibitor that mainly inhibits neutrophil elastase in the lungs. A variant of A1AT at the P1 position with methionine 358 to arginine (A1AT-Pittsburgh) is a rapid inhibitor of thrombin with greatly diminished anti-elastase activity. The P2 residue (position 357) of A1AT-Pittsburgh has been shown to play an important role in interactions with thrombin and kallikrein, but the role of P2 residue in wild-type A1AT has largely been unraveled. Here, we investigated the effects of P2 proline substitutions in wild-type A1AT on interactions with porcine pancreatic elastase (PPE) and human neutrophil elastase (HNE)...
2017: PloS One
https://www.readbyqxmd.com/read/28919686/high-performance-liquid-chromatography-ultraviolet-photodiode-array-detection-method-for-aflatoxin-b1-in-cattle-feed-supplements
#2
Lazuardi Mochamad, Bambang Hermanto
AIM: The objective of the current study is to determine the concentration of aflatoxin B1 using high-performance liquid chromatography (HPLC) with a photodiode array (PDA) detector. MATERIALS AND METHODS: Aflatoxin B1 certified reference grade from Trilogy Analytical Laboratory dissolved acetonitrile (ACN) at 10 µg/mL was using standard assessment. HPLC instruments such as ultraviolet-PDA detector used a Shimadzu LC-6AD pump with DGU-20A5 degasser, communication module-20A, and PDA detector SPD-M20A with FRC-10A fraction collector...
August 2017: Veterinary World
https://www.readbyqxmd.com/read/28918898/endoplasmic-reticulum-transport-of-glutathione-by-sec61-is-regulated-by-ero1-and-bip
#3
Alise J Ponsero, Aeid Igbaria, Maxwell A Darch, Samia Miled, Caryn E Outten, Jakob R Winther, Gael Palais, Benoit D'Autréaux, Agnès Delaunay-Moisan, Michel B Toledano
In the endoplasmic reticulum (ER), Ero1 catalyzes disulfide bond formation and promotes glutathione (GSH) oxidation to GSSG. Since GSSG cannot be reduced in the ER, maintenance of the ER glutathione redox state and levels likely depends on ER glutathione import and GSSG export. We used quantitative GSH and GSSG biosensors to monitor glutathione import into the ER of yeast cells. We found that glutathione enters the ER by facilitated diffusion through the Sec61 protein-conducting channel, while oxidized Bip (Kar2) inhibits transport...
September 11, 2017: Molecular Cell
https://www.readbyqxmd.com/read/28917942/in-vivo-and-in-vitro-testing-of-native-%C3%AE-conotoxins-from-the-injected-venom-of-conus-purpurascens
#4
REVIEW
Mickelene F Hoggard, Alena M Rodriguez, Herminsul Cano, Evan Clark, Han-Shen Tae, David J Adams, Tanja A Godenschwege, Frank Marí
α-Conotoxins inhibit nicotinic acetylcholine receptors (nAChRs) and are used as probes to study cholinergic pathways in vertebrates. Model organisms, such as Drosophila melanogaster, express nAChRs in their CNS that are suitable to investigate the neuropharmacology of α-conotoxins in vivo. Here we report the paired nanoinjection of native α-conotoxin PIA and two novel α-conotoxins, PIC and PIC[O7], from the injected venom of Conus purpurascens and electrophysiological recordings of their effects on the giant fiber system (GFS) of D...
September 13, 2017: Neuropharmacology
https://www.readbyqxmd.com/read/28916383/heterologous-expression-and-structure-function-relationship-of-low-temperature-and-alkaline-active-protease-from-acinetobacter-sp-ihb-b-5011-mn12
#5
Richa Salwan, Vivek Sharma, Mohinder Pal, Ramesh Chand Kasana, Sudesh Kumar Yadav, Arvind Gulati
The gene encoding protease from Acinetobacter sp. IHB B 5011(MN12) was cloned and expressed in Escherichia coli BL21(DE3). The nucleotide sequence revealed 1323bp ORF encoding 441 amino acids protein with molecular weight 47.2kDa. The phylogenetic analysis showed clustering of Alp protease with subtilisin-like serine proteases of S8 family. The amino acid sequence was comprised of N-terminal signal peptide 1-21 amino acids, pre-peptide 22-143 amino acids, peptidase S8 domain 144-434 amino acids, and pro-peptide 435-441 amino acids at C-terminus...
September 12, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28912346/the-blood-pressure-lowering-effect-of-20-hete-blockade-in-cyp4a14-mice-is-associated-with-natriuresis
#6
Varunkumar Pandey, Victor Garcia, Ankit Gilani, Priyanka Mishra, Frank Fan Zhang, Mahesh P Paudyal, John R Falck, Alberto Nasjletti, Wen-Hui Wang, Michal L Schwartzman
20-Hydroxy-5,8,11,14-eicosatetraenoic acid (20-HETE) has been linked to pro- and anti-hypertensive actions through its ability to promote vasoconstriction and inhibit Na transport in the ascending limb of the loop of Henle, respectively. In this study, we assessed the effects of 20-HETE blockade on blood pressure, renal hemodynamics and urinary sodium excretion in Cyp4a14(-/-) male mice, which display androgen-driven 20-HETE-dependent hypertension. Administration of 20-SOLA, a water-soluble 20-HETE antagonist, in the drinking water normalized blood pressure of male Cyp4a14(-/-) hypertensive mice (124± vs 153± mmHg) while having no effect on age-matched normotensive wild-type male mice...
September 14, 2017: Journal of Pharmacology and Experimental Therapeutics
https://www.readbyqxmd.com/read/28911360/evaluation-of-the-tb-lamp-assay-for-the-rapid-diagnosis-of-pulmonary-tuberculosis-in-northern-india
#7
R Yadav, N Sharma, R Khaneja, P Agarwal, A Kanga, D Behera, S Sethi
SETTING: A tertiary care hospital in North India. OBJECTIVE: To evaluate a commercial kit-based loop-mediated isothermal amplification (TB-LAMP) assay for the diagnosis of pulmonary tuberculosis (PTB). DESIGN: A total of 530 patients presenting with PTB symptoms were enrolled and one sputum sample was collected from each patient. The TB-LAMP assay (Loopamp™ MTBC Detection kit) was performed on the raw sputum sample. The remaining sample was used for smear microscopy and mycobacterial culture...
October 1, 2017: International Journal of Tuberculosis and Lung Disease
https://www.readbyqxmd.com/read/28910577/subnuclear-distribution-of-proteins-links-with-genome-architecture
#8
Fouziya R Shah, Younus A Bhat, Ajazul H Wani
Metazoan genomes have a hierarchal 3-dimensional (3D) organization scaling from nucleosomes, loops, topologically associating domains (TADs), compartments, to chromosome territories. The 3D organization of genome has been linked with development, differentiation and disease. However, the principles governing the 3D chromatin architecture are just beginning to get unraveled. The nucleus has very high concentration of proteins and these proteins are either diffusely distributed throughout the nucleus, or aggregated in the form of foci/bodies/clusters/speckles or in combination of both...
September 14, 2017: Nucleus
https://www.readbyqxmd.com/read/28905451/gene-edited-human-kidney-organoids-reveal-mechanisms-of-disease-in-podocyte-development
#9
Yong Kyun Kim, Ido Refaeli, Craig R Brooks, Peifeng Jing, Ramila E Gulieva, Michael R Hughes, Nelly M Cruz, Yannan Liu, Angela J Churchill, Yuliang Wang, Hongxia Fu, Jeffrey W Pippin, Lih Y Lin, Stuart J Shankland, A Wayne Vogl, Kelly M McNagny, Benjamin S Freedman
A critical event during kidney organogenesis is the differentiation of podocytes, specialized epithelial cells that filter blood plasma to form urine. Podocytes derived from human pluripotent stem cells (hPSC-podocytes) have recently been generated in nephron-like kidney organoids, but the developmental stage of these cells and their capacity to reveal disease mechanisms remains unclear. Here we show that hPSC-podocytes phenocopy mammalian podocytes at the capillary loop stage (CLS), recapitulating key features of ultrastructure, gene expression, and mutant phenotype...
September 14, 2017: Stem Cells
https://www.readbyqxmd.com/read/28900596/a-review-of-demodulation-techniques-for-amplitude-modulation-atomic-force-microscopy
#10
REVIEW
Michael G Ruppert, David M Harcombe, Michael R P Ragazzon, S O Reza Moheimani, Andrew J Fleming
In this review paper, traditional and novel demodulation methods applicable to amplitude-modulation atomic force microscopy are implemented on a widely used digital processing system. As a crucial bandwidth-limiting component in the z-axis feedback loop of an atomic force microscope, the purpose of the demodulator is to obtain estimates of amplitude and phase of the cantilever deflection signal in the presence of sensor noise or additional distinct frequency components. Specifically for modern multifrequency techniques, where higher harmonic and/or higher eigenmode contributions are present in the oscillation signal, the fidelity of the estimates obtained from some demodulation techniques is not guaranteed...
2017: Beilstein Journal of Nanotechnology
https://www.readbyqxmd.com/read/28900561/effects-of-a-band-loop-on-lower-extremity-muscle-activity-and-kinematics-during-the-barbell-squat
#11
Ryan C A Foley, Brittany D Bulbrook, Duane C Button, Michael W R Holmes
BACKGROUND: Medial knee collapse can signal an underlying movement issue that, if uncorrected, can lead to a variety of knee injuries. Placing a band around the distal thigh may act as a proprioceptive aid to minimize medial collapse of the knee during squats; however, little is known about EMG and biomechanics in trained and untrained individuals during the squat with an elastic band added. HYPOTHESIS/PURPOSE: To investigate the effects of the TheraBand® Band Loop on kinematics and muscle activity of the lower extremity during a standard barbell back squat at different intensities in both trained and untrained individuals...
August 2017: International Journal of Sports Physical Therapy
https://www.readbyqxmd.com/read/28900197/structural-insights-into-the-middle-east-respiratory-syndrome-coronavirus-4a-protein-and-its-dsrna-binding-mechanism
#12
Maria Batool, Masaud Shah, Mahesh Chandra Patra, Dhanusha Yesudhas, Sangdun Choi
Middle East respiratory syndrome coronavirus (MERS-CoV) has evolved to navigate through the sophisticated network of a host's immune system. The immune evasion mechanism including type 1 interferon and protein kinase R-mediated antiviral stress responses has been recently attributed to the involvement of MERS-CoV protein 4a (p4a) that masks the viral dsRNA. However, the structural mechanism of how p4a recognizes and establishes contacts with dsRNA is not well explained. In this study, we report a dynamic mechanism deployed by p4a to engage the viral dsRNA and make it unavailable to the host immune system...
September 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28892380/gne-781-a-highly-advanced-potent-and-selective-bromodomain-inhibitor-of-cyclic-adenosine-monophosphate-response-element-binding-protein-binding-protein-cbp
#13
F Anthony Romero, Jeremy M Murray, Kwong Wah Lai, Vickie Tsui, Brian K Albrecht, Le An, Maureen H Beresini, Gladys de Leon Boenig, Sarah M Bronner, Emily W Chan, Kevin X Chen, Zhongguo Chen, Edna F Choo, Kyle Clagg, Kevin Clark, Terry D Crawford, Patrick Cyr, Denise De Almeida Nagata, Karen E Gascoigne, Jane L Grogan, Georgia Hatzivassiliou, Wei Huang, Thomas L Hunsaker, Susan Kaufman, Stefan G Koenig, Ruina Li, Yingjie Li, Xiaorong Liang, Jiangpeng Liao, Wenfeng Liu, Justin Q Ly, Jonathan Maher, Colin Masui, Mark Merchant, Yingqing Ran, Alexander M Taylor, John S Wai, Fei Wang, Xiaocang Wei, Dong Yu, Bing-Yan Zhu, Xiaoyu Zhu, Steven R Magnuson
Inhibition of the bromodomain of the transcriptional regulator CBP/P300 is an especially interesting new therapeutic approach in oncology. We recently disclosed in vivo chemical tool 1 (GNE-272) for the bromodomain of CBP that was moderately potent and selective over BRD4(1). In pursuit of a more potent and selective CBP inhibitor we used structure-based design. Constraining the aniline of 1 into a tetrahydroquinoline motif maintained potency and increased selectivity 2-fold. Structure-activity relationship studies coupled with further structure-based design targeting the LPF shelf, BC loop and KAc regions allowed us to significantly increase potency and selectivity, resulting in the identification of non-CNS penetrant 19 (GNE-781; TR-FRET IC50 = 0...
September 11, 2017: Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28891390/pathology-and-epidemiology-of-oxalate-nephrosis-in-cheetahs
#14
Emily P Mitchell, Molly E Church, Sarah M Nemser, Betsy Jean Yakes, Eric R Evans, Renate Reimschuessel, Karin Lemberger, Peter N Thompson, Karen A Terio
To investigate cases of acute oxalate nephrosis without evidence of ethylene glycol exposure, archived data and tissues from cheetahs ( Acinonyx jubatus) from North America ( n = 297), southern Africa ( n = 257), and France ( n = 40) were evaluated. Renal and gastrointestinal tract lesions were characterized in a subset of animals with ( n = 100) and without ( n = 165) oxalate crystals at death. Crystals were confirmed as calcium oxalate by Raman spectroscopy in 45 of 47 cheetahs tested. Crystals were present in cheetahs from 3...
January 1, 2017: Veterinary Pathology
https://www.readbyqxmd.com/read/28887323/atg5-mediates-a-positive-feedback-loop-between-wnt-signaling-and-autophagy-in-melanoma
#15
Abibatou Ndoye, Anna Budina-Kolomets, Curtis H Kugel, Marie R Webster, Amanpreet Kaur, Reeti Behera, Vito W Rebecca, Ling Li, Patricia A Brafford, Qin Liu, Y N Vashisht Gopal, Michael A Davies, Gordon B Mills, Xiaowei Xu, Hong Wu, Meenhard Herlyn, Michael C Nicastri, Jeffrey D Winkler, Maria S Soengas, Ravi K Amaravadi, Maureen E Murphy, Ashani T Weeraratna
Autophagy mediates resistance to various anticancer agents. In melanoma, resistance to targeted therapy has been linked to expression of Wnt5A, an intrinsic inhibitor of β-catenin, which also promotes invasion. In this study, we assessed the interplay between Wnt5A and autophagy by combining expression studies in human clinical biopsies with functional analyses in cell lines and mouse models. Melanoma cells with high Wnt5A and low β-catenin displayed increased basal autophagy. Genetic blockade of autophagy revealed an unexpected feedback loop whereby knocking down the autophagy factor ATG5 in Wnt5Ahigh cells decreased Wnt5A and increased β-catenin...
September 8, 2017: Cancer Research
https://www.readbyqxmd.com/read/28885628/fluctuation-tension-and-shape-transition-of-vesicles-renormalisation-calculations-and-monte-carlo-simulations
#16
Guillaume Gueguen, Nicolas Destainville, Manoel Manghi
It has been known for long that the fluctuation surface tension of membranes r, computed from the height fluctuation spectrum, is not equal to the bare surface tension σ, which is introduced in the theory either as a Lagrange multiplier to conserve the total membrane area or as an external constraint. In this work we relate these two surface tensions both analytically and numerically. They are also compared to the Laplace tension γ, and the mechanical frame tension τ. Using the Helfrich model and one-loop renormalisation calculations, we obtain, in addition to the effective bending modulus κeff, a new expression for the effective surface tension σeff = σ - εkBT/(2ap) where kBT is the thermal energy, ap the projected cut-off area, and ε = 3 or 1 according to the allowed configurations that keep either the projected area or the total area constant...
September 8, 2017: Soft Matter
https://www.readbyqxmd.com/read/28882898/switch-i-dependent-allosteric-signaling-in-a-g-protein-chaperone-b12-enzyme-complex
#17
Gregory C Campanello, Michael Lofgren, Adam L Yokom, Daniel R Southworth, Ruma Banerjee
G-proteins regulate various processes ranging from DNA replication and protein synthesis to cytoskeletal dynamics and cofactor assimilation, and serve as models for uncovering strategies deployed for allosteric signal transduction. MeaB is a multifunctional G-protein chaperone, which gates loading of the active 5'-deoxyadenosylcobalamin cofactor onto methylmalonyl-CoA mutase (MCM) and precludes loading of inactive cofactor forms. MeaB also safeguards MCM, which uses radical chemistry, against inactivation and rescues MCM inactivated during catalytic turnover by using the GTP-binding energy to offload inactive cofactor...
September 7, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28882005/cats-coordinates-of-atoms-by-taylor-series-protein-design-with-backbone-flexibility-in-all-locally-feasible-directions
#18
Mark A Hallen, Bruce R Donald
Motivation: When proteins mutate or bind to ligands, their backbones often move significantly, especially in loop regions. Computational protein design algorithms must model these motions in order to accurately optimize protein stability and binding affinity. However, methods for backbone conformational search in design have been much more limited than for sidechain conformational search. This is especially true for combinatorial protein design algorithms, which aim to search a large sequence space efficiently and thus cannot rely on temporal simulation of each candidate sequence...
July 15, 2017: Bioinformatics
https://www.readbyqxmd.com/read/28880281/a-right-handed-signalling-pathway-drives-heart-looping-in-vertebrates
#19
Oscar H Ocaña, Hakan Coskun, Carolina Minguillón, Prayag Murawala, Elly M Tanaka, Joan Galcerán, Ramón Muñoz-Chápuli, M Angela Nieto
Most animals show external bilateral symmetry, which hinders the observation of multiple internal left-right (L/R) asymmetries that are fundamental to organ packaging and function. In vertebrates, left identity is mediated by the left-specific Nodal-Pitx2 axis that is repressed on the right-hand side by the epithelial-mesenchymal transition (EMT) inducer Snail1 (refs 3, 4). Despite some existing evidence, it remains unclear whether an equivalent instructive pathway provides right-hand-specific information to the embryo...
September 6, 2017: Nature
https://www.readbyqxmd.com/read/28878352/s1pr1-drives-a-feed-forward-signalling-loop-to-regulate-batf3-and-the-transcriptional-programme-of-hodgkin-lymphoma-cells
#20
K Vrzalikova, M Ibrahim, M Vockerodt, T Perry, S Margielewska, L Lupino, E Nagy, E Soilleux, D Liebelt, R Hollows, A Last, G Reynolds, M Abdullah, H Curley, M Care, D Krappmann, R Tooze, J Allegood, S Spiegel, W Wei, C B J Woodman, P G Murray
The Hodgkin/Reed-Sternberg (HRS) cells of classical Hodgkin lymphoma (HL) are characterised by the aberrant activation of multiple signalling pathways. Here we show that a subset of HL displays altered expression of sphingosine-1-phosphate (S1P) receptors (S1PR). S1P activates phosphatidylinositide 3-kinase (PI3-K) in these cells that is mediated by the increased expression of S1PR1 and the decreased expression of S1PR2. We also showed that genes regulated by PI3-K signalling pathway in HL cell lines significantly overlap with the transcriptional programme of primary HRS cells...
September 7, 2017: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
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