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https://www.readbyqxmd.com/read/29684909/hyphenation-of-size-exclusion-chromatography-to-native-ion-mobility-mass-spectrometry-for-the-analytical-characterization-of-therapeutic-antibodies-and-related-products
#1
Anthony Ehkirch, Oscar Hernandez-Alba, Olivier Colas, Alain Beck, Davy Guillarme, Sarah Cianférani
Mass spectrometry performed in non-denaturing conditions (native MS), and its hyphenation to ion mobility spectrometry (IM-MS), have gained interest for the qualitative and quantitative characterization of intact monoclonal antibody-related (mAb) products. However, one main drawback is the manual sample preparation, which hampers its routine use in high throughput automated environments. Size exclusion chromatography (SEC) appears as an interesting technique to perform online buffer exchange in an automated way...
April 12, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/29684813/metabolite-characterization-of-ambrisentan-in-in-vitro-and-in-vivo-matrices-by-uhplc-qtof-ms-ms-detection-of-glutathione-conjugate-of-epoxide-metabolite-evidenced-by-in-vitro-gsh-trapping-assay
#2
P Johnsi Rani, Chiguru Vishnuvardhan, Rakesh D Nimbalkar, Prabha Garg, N Satheeshkumar
The focus of the present study is on in vitro and in vivo metabolite identification of ambrisentan (AMBR) a selective endothelin type - A (ETA) receptor antagonist using quadruple time-of-flight mass spectrometry (QTOF/MS). in vitro metabolism study was conducted by incubating AMBR in rat liver microsomes (RLM), rat and human liver S9 fractions. In vivo study was carried out through the collection of urine, faeces and plasma samples at various time points after oral administration of AMBR in suspension form at a dose of 25 mg/kg to six male Sprague - Dawley (SD) rats...
April 12, 2018: Journal of Pharmaceutical and Biomedical Analysis
https://www.readbyqxmd.com/read/29684785/photoreceptor-calcium-sensor-proteins-in-detergent-resistant-membrane-rafts-are-regulated-via-binding-to-caveolin-1
#3
Vasiliy I Vladimirov, Evgeni Yu Zernii, Viktoriia E Baksheeva, Hanna Wimberg, Alexey S Kazakov, Natalya K Tikhomirova, Ekaterina L Nemashkalova, Vladimir A Mitkevich, Andrey A Zamyatnin, Valery M Lipkin, Pavel P Philippov, Sergei E Permyakov, Ivan I Senin, Karl-W Koch, Dmitry V Zinchenko
Rod cell membranes contain cholesterol-rich detergent-resistant membrane (DRM) rafts, which accumulate visual cascade proteins as well as proteins involved in regulation of phototransduction such as rhodopsin kinase and guanylate cyclases. Caveolin-1 is the major integral component of DRMs, possessing scaffolding and regulatory activities towards various signaling proteins. In this study, photoreceptor Ca2+ -binding proteins recoverin, NCS1, GCAP1, and GCAP2, belonging to neuronal calcium sensor (NCS) family, were recognized as novel caveolin-1 interacting partners...
April 14, 2018: Cell Calcium
https://www.readbyqxmd.com/read/29684759/using-structurally-defined-oligosaccharides-to-understand-the-interactions-between-proteins-and-heparan-sulfate
#4
REVIEW
Ding Xu, Katelyn Arnold, Jian Liu
Heparan sulfate (HS) is widely present on the animal cell surface and in the extracellular matrix. HS achieves its biological functions by interacting with proteins to change proteins' conformation, oligomerization state and cellular location. The challenging question to study HS is how to dissect the relationship between the structures of HS and the biological activities. In the past several years, crucial techniques have been developed to overcome this challenge. A novel chemoenzymatic method to synthesize structurally defined HS oligosaccharides has offered a key access to this class of sulfated carbohydrate molecules...
April 20, 2018: Current Opinion in Structural Biology
https://www.readbyqxmd.com/read/29684709/the-role-of-natural-selection-in-shaping-genetic-variation-in-a-promising-chagas-disease-drug-target-trypanosoma-cruzi-trans-sialidase
#5
Joseph P Gallant, Raquel Asunción Lima-Cordón, Silvia Justi, Maria Carlota Monroy, Toni Viola, Lori Stevens
Rational drug design creates innovative therapeutics based on knowledge of the biological target to provide more effective and responsible therapeutics. Chagas disease, endemic throughout Latin America, is caused by Trypanosoma cruzi, a protozoan parasite. Current therapeutics are problematic with widespread calls for new approaches. Researchers are using rational drug design for Chagas disease and one target receiving considerable attention is the T. cruzi trans-sialidase protein (TcTS). In T. cruzi, trans-sialidase catalyzes the transfer of sialic acid from a mammalian host to coat the parasite surface membrane and avoid immuno-detection...
April 20, 2018: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/29684708/structural-based-design-synthesis-and-antitumor-activity-of-novel-alloxazine-analogues-with-potential-selective-kinase-inhibition
#6
Waleed H Malki, Ahmed M Gouda, Hamdy E A Ali, Rabaa Al-Rousan, Doaa Samaha, Ashraf N Abdalla, Juan Bustamante, Zakaria Y Abd Elmageed, Hamed I Ali
Protein kinases are promising therapeutic targets for cancer therapy. Here, we applied multiple approaches to optimize the potency and selectivity of our reported alloxazine scaffold. Flexible moieties at position 2 of the hetero-tricyclic system were incorporated to fit into the ATP binding site and extend to the adjacent allosteric site and selectively inhibit protein kinases. This design led to potential selective inhibition of ABL1, CDK1/Cyclin A1, FAK, and SRC kinase by 30-59%. Cytotoxicity was improved by ∼50 times for the optimized lead (10b; IC50  = 40 nM) against breast cancer (MCF-7) cells...
April 17, 2018: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/29684677/human-stem-cell-based-corneal-tissue-mimicking-structures-using-laser-assisted-3d-bioprinting-and-functional-bioinks
#7
Anni Sorkio, Lothar Koch, Laura Koivusalo, Andrea Deiwick, Susanna Miettinen, Boris Chichkov, Heli Skottman
There is a high demand for developing methods to produce more native-like 3D corneal structures. In the present study, we produced 3D cornea-mimicking tissues using human stem cells and laser-assisted bioprinting (LaBP). Human embryonic stem cell derived limbal epithelial stem cells (hESC-LESC) were used as a cell source for printing epithelium-mimicking structures, whereas human adipose tissue derived stem cells (hASCs) were used for constructing layered stroma-mimicking structures. The development and optimization of functional bioinks was a crucial step towards successful bioprinting of 3D corneal structures...
April 16, 2018: Biomaterials
https://www.readbyqxmd.com/read/29684597/comparative-chloroplast-genomics-between-the-invasive-weed-mikania-micrantha-and-its-indigenous-congener-mikania-cordata-structure-variation-identification-of-highly-divergent-regions-divergence-time-estimation-and-phylogenetic-analysis
#8
Yingjuan Su, Lu Huang, Zhen Wang, Ting Wang
Mikania micrantha and Mikania cordata are the only two species in genus Mikania (Asteraceae) in China. They share very similar morphological and life-history characteristics but occupy quite different habitats. Most importantly, they generate totally different ecological consequences. While M. micrantha has become an exotic invasive weed, M. cordata exists as an indigenous species with no harmful effects on native plants or habitats. As a continuous study of our previously reported M. micrantha chloroplast (cp) genome, in this study we have further sequenced the M...
April 20, 2018: Molecular Phylogenetics and Evolution
https://www.readbyqxmd.com/read/29684584/deletion-of-epithelial-cell-specific-cdc42-leads-to-enamel-hypermaturation-in-a-conditional-knockout-mouse-model
#9
Zhihui Tian, Xiaolin Lv, Min Zhang, Xueer Wang, Yinghua Chen, Pei Tang, Pengcheng Xu, Lu Zhang, Buling Wu, Lin Zhang
Recent evidence suggests that GTPases Rho family plays an important role in tooth development; however, the role of Cdc42 in tooth development remains unclear. We aimed to investigate the function of Cdc42 in tooth development and amelogenesis. We generated an epithelial cell-specific K5-Cdc42 knockout (KO) mouse to evaluate post-eruption dental phenotypes using a K5-Cre driver line. This model overcomes the previously reported perinatal lethality. Tooth phenotypes were analyzed by micro X-ray, micro-computed tomography (CT), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), wear rate, shear strength, and a microhardness test...
April 20, 2018: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29684538/molecular-cloning-characterization-and-expression-analysis-of-c-ebp-%C3%AE-%C3%AE-and-%C3%AE-in-adipose-related-tissues-and-adipocyte-of-duck-anas-platyrhynchos
#10
Jiamin Qiu, Wanxia Wang, Shenqiang Hu, Yushi Wang, Wenqiang Sun, Jiwei Hu, Jiwen Wang
CCAAT/enhancer binding protein α, β, δ (C/EBP α, β, δ) are essential transcriptional factors in regulating adipose development. However, information about their sequence characteristics and functions during adipocyte development still remains scarce in birds. In present study, we found that duck C/EBP α, β, δ differed in their phosphorylation sites and low complexity regions (LCRs) among their orthologs and paralogs. Phylogenetic analysis showed that C/EBP α, β, δ had different evolutionary patterns, and each of duck C/EBP α, β, δ was strikingly diverged from orthologs of other Aves...
April 20, 2018: Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
https://www.readbyqxmd.com/read/29684533/lipid-coated-mannitol-core-microparticles-for-sustained-release-of-protein
#11
Bifeng Wang, Wolfgang Friess
Parenteral sustained release systems for proteins which provide therapeutic levels over a longer period avoiding frequent administration, which preserve protein stability during manufacturing, storage and application and which are biodegradable and highly biocompatible in the body are intensively sought after. The aim of this study was to generate and study mannitol core microparticles loaded with a monoclonal antibody IgG1 and coated with lipid either hard fat or glyceryl stearate at different coating levels...
April 20, 2018: European Journal of Pharmaceutics and Biopharmaceutics
https://www.readbyqxmd.com/read/29684512/functional-characterization-and-molecular-modeling-of-the-mutations-in-cyp21a2-gene-from-patients-with-congenital-adrenal-hyperplasia
#12
Ragini Khajuria, Rama Walia, Anil Bhansali, Rajendra Prasad
Steroid 21-Hydroxylase deficiency is an inherited autosomal recessive metabolic disorder of the adrenal steroidogenesis caused due to mutations in the CYP21A2 gene in 95% of CAH cases. Notably, the de novo mutations arise at the rate of 3-5%, therefore the functional characterization is of utmost importance for categorization of the novel mutations. Herein, we have functionally characterized the CYP21A2 missense mutations viz., p. F306V and p. H365 N. Notably, both the mutations were harbored by the patients exhibiting the non classical phenotype...
April 20, 2018: Biochimie
https://www.readbyqxmd.com/read/29684511/the-effect-of-oxysterols-on-nerve-impulses
#13
REVIEW
Maryem Bezine, Amira Namsi, Randa Sghaier, Rym Ben Khalifa, Haithem Hamdouni, Fatiha Brahmi, Iham Badreddine, Wafa Mihoubi, Thomas Nury, Anne Vejux, Amira Zarrouk, Jérôme de Sèze, Thibault Moreau, Boubker Nasser, Gérard Lizard
The propagation of nerve impulses in myelinated nerve fibers depends on a number of factors involving the myelin and neural axons. In several neurodegenerative diseases, nerve impulses can be affected by the structural and biochemical characteristics of the myelin sheath and the activity of ion channels located in the nodes of Ranvier. Though it is generally accepted that lipid disorders are involved in the development of neurodegenerative diseases, little is known about their impact on nerve impulses. Cholesterol oxide derivatives (also called oxysterols), which are either formed enzymatically or as a result of cholesterol auto-oxidation or both, are often found in abnormal levels in the brain and body fluids of patients with neurodegenerative diseases...
April 20, 2018: Biochimie
https://www.readbyqxmd.com/read/29684503/dynamics-based-community-analysis-and-perturbation-response-scanning-of-allosteric-interaction-networks-in-the-trap1-chaperone-structures-dissect-molecular-linkage-between-conformational-asymmetry-and-sequential-atp-hydrolysis
#14
Gennady M Verkhivker
Allosteric interactions of the Hsp90 chaperones with cochaperones and diverse protein clients can often exhibit distinct asymmetric features that determine regulatory mechanisms and cellular functions in many signaling networks. The recent crystal structures of the mitochondrial Hsp90 isoform TRAP1 in complexes with ATP analogs have provided first evidence of significant asymmetry in the closed dimerized state that triggers independent activity of the chaperone protomers, whereby preferential hydrolysis of the buckled protomer is followed by conformational flipping between protomers and hydrolysis of the second protomer...
April 20, 2018: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29684483/three-dimensional-protein-shape-similarity-analysis-based-on-hybrid-features
#15
Zhong Li, Jiangjiang Yu, Hailong Hu, Shuiwang Ji
The analysis of protein similarity is a matter of concern in the bioinformatics field, since studying the protein similarity can help understand the protein structure-function relationship. To this aim, several methods have been proposed, but currently, protein similarity results are still not satisfactory. Here we presented a novel method for evaluating the similarity of 3D protein models based on hybrid features, including the local diameter (LD), the salient geometric feature (SGF) and the heat kernel signature (HKS)...
April 20, 2018: Gene
https://www.readbyqxmd.com/read/29684466/the-g-protein-coupled-receptor-gpr34-the-past-20-years-of-a-grownup
#16
REVIEW
Torsten Schöneberg, Jaroslawna Meister, Alexander Bernd Knierim, Angela Schulz
Research on GPR34, which was discovered in 1999 as an orphan G protein-coupled receptor of the rhodopsin-like class, disclosed its physiologic relevance only piece by piece. Being present in all recent vertebrate genomes analyzed so far it seems to improve the fitness of species although it is not essential for life and reproduction as GPR34-deficient mice demonstrate. However, closer inspection of macrophages and microglia, where it is mainly expressed, revealed its relevance in immune cell function. Recent data clearly demonstrate that GPR34 function is required to arrest microglia in the M0 homeostatic non-phagocytic phenotype...
April 20, 2018: Pharmacology & Therapeutics
https://www.readbyqxmd.com/read/29684428/validation-of-a-cd1b-tetramer-assay-for-studies-of-human-mycobacterial-infection-or-vaccination
#17
Erik D Layton, Krystle K Q Yu, Malisa T Smith, Thomas J Scriba, Stephen C De Rosa, Chetan Seshadri
CD1 tetramers loaded with lipid antigens facilitate the identification of rare lipid-antigen specific T cells present in human blood and tissue. Because CD1 proteins are structurally non-polymorphic, these tetramers can be applied to genetically diverse human populations, unlike MHC-I and MHC-II tetramers. However, there are no standardized assays to quantify and characterize lipid antigen-specific T cells present within clinical samples. We incorporated CD1b tetramers loaded with the mycobacterial lipid glucose monomycolate (GMM) into a multi-parameter flow cytometry assay...
April 20, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29684416/development-of-antibody-biomarkers-of-long-term-and-recent-dengue-virus-infections
#18
Eduardo J M Nascimento, James W Huleatt, Marli T Cordeiro, Priscila M S Castanha, James K George, Eduard Grebe, Alex Welte, Monique Brown, Donald S Burke, Ernesto T A Marques
Dengue virus (DENV) infections elicit antibody responses to the non-structural protein 1 (NS1) that are associated with protection against disease. However, the antibody isotypes and subclasses involved, and their kinetics have not been extensively studied. We characterized the antibody responses to DENV NS1 by enzyme-linked immunosorbent assay (ELISA) in a longitudinal cohort of 266 confirmed dengue cases in Recife, Northeast Brazil. Samples were collected during the febrile phase and up to over 3 years after onset of symptoms...
April 20, 2018: Journal of Virological Methods
https://www.readbyqxmd.com/read/29684376/angiotensin-ii-cyclic-analogs-as-tools-to-investigate-at-1-r-biased-signaling-mechanisms
#19
David St-Pierre, Jérôme Cabana, Brian J Holleran, Élie Besserer-Offroy, Emanuel Escher, Gaétan Guillemette, Pierre Lavigne, Richard Leduc
G protein coupled receptors (GPCRs) produce pleiotropic effects by their capacity to engage numerous signaling pathways once activated. Functional selectivity (also called biased signaling), where specific compounds can bring GPCRs to adopt conformations that enable selective receptor coupling to distinct signaling pathways, continues to be significantly investigated. However, an important but often overlooked aspect of functional selectivity is the capability of ligands such as angiotensin II (AngII) to adopt specific conformations that may preferentially bind to selective GPCRs structures...
April 20, 2018: Biochemical Pharmacology
https://www.readbyqxmd.com/read/29684364/inhibition-of-mitochondrial-fission-preserves-photoreceptors-after-retinal-detachment
#20
Xiangjun She, Xinmin Lu, Tong Li, Junran Sun, Jian Liang, Yuanqi Zhai, Shiqi Yang, Qing Gu, Fang Wei, Hong Zhu, Fenghua Wang, Xueting Luo, Xiaodong Sun
Photoreceptor degeneration is a leading cause of visual impairment worldwide. Separation of neurosensory retina from the underlying retinal pigment epithelium is a prominent feature preceding photoreceptor degeneration in a variety of retinal diseases. Although ophthalmic surgeries have been well developed to restore retinal structures, post-op patients usually experience progressive photoreceptor degeneration and irreversible vision loss that is incurable at present. Previous studies point to a critical role of mitochondria-mediated apoptotic pathway in photoreceptor degeneration, but the upstream triggers remain largely unexplored...
April 20, 2018: American Journal of Pathology
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