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Assay development

Zheng-Zheng Zhang, Yun-Feng Zhou, Wei-Ping Li, Chuan Jiang, Zhong Chen, Huan Luo, Bin Song
BACKGROUND: Although many strategies have been developed to modify the biological and biomechanical environment of the meniscal suture repair to improve the chances of healing, the failure rates remain high. Thus, new methods to promote meniscal regeneration and repair are needed. HYPOTHESIS: Administration of magnesium (via a repair using magnesium stitches) might enhance recruitment and adherence of endogenous stem cells to the site of the lesion, thereby promoting in situ meniscal regeneration and chondroprotective functions...
February 20, 2019: American Journal of Sports Medicine
Dirk Lindenblatt, Mareike Horn, Claudia Götz, Karsten Niefind, Ines Neundorf, Markus Pietsch
The ubiquitously expressed Ser/Thr kinase CK2 is a key regulator in a variety of key processes in normal and malignant cells. Due to its distinctive anti-apoptotic and tumor-driving properties, elevated levels of CK2 have frequently been found in tumors of different origin. In recent years, development of CK2 inhibitors has largely been focused on ATP-competitive compounds; however, targeting the CK2α/CK2β interface has emerged as a further concept that might avoid selectivity issues. In order to address the CK2 subunit interaction site, we have synthesized halogenated CK2β-mimicking cyclic peptides modified with the cell-penetrating peptide sC18 to mediate cellular uptake...
February 20, 2019: ChemMedChem
Jianming Wu, Hemant K Mishra, Bruce Walcheck
Human NK cell antitumor activities involve Ab-dependent cell-mediated cytotoxicity (ADCC), which is a key mechanism of action for several clinically successful tumor-targeting therapeutic mAbs. Human NK cells exclusively recognize these Abs by the Fcγ receptor CD16A (FcγRIIIA), one of their most potent activating receptors. Unlike other activating receptors on NK cells, CD16A undergoes a rapid down-regulation in expression by a proteolytic process following NK cell activation with various stimuli. In this review, the role of a disintegrin and metalloproteinase-17 (ADAM17) in CD16A cleavage and as a regulatory checkpoint is discussed...
February 20, 2019: Journal of Leukocyte Biology
Yvonne J Rosenberg, George K Lewis, David C Montefiori, Celia C LaBranche, Mark G Lewis, Lori A Urban, Jonathan P Lees, Lingjun Mao, Xiaoming Jiang
Recombinant antibodies play increasingly important roles as immunotherapeutic treatments for human cancers as well as inflammatory and infectious diseases and have revolutionized their management. In addition, their therapeutic potential may be enhanced by the introduction of defined mutations in the crystallizable fragment (Fc) domains eg YTE (M252Y/S254T/T256E) and LS (M428L/N434S), as a consequence of increased half-lives and prolonged duration of protection. However, the functional properties of any biologic may be compromised by unanticipated immunogenicity in humans, rendering them ineffective...
2019: PloS One
Nitin Agrawal, Soili I Lehtonen, Meri Uusi-Mäkelä, Purvi Jain, Sari Viitala, Juha A E Määttä, Niklas Kähkönen, Latifeh Azizi, Tiina A Riihimäki, Markku S Kulomaa, Mark S Johnson, Vesa P Hytönen, Tomi T Airenne
Chicken avidin (Avd) and streptavidin from Streptomyces avidinii are extensively used in bionanotechnology due to their extremely tight binding to biotin (Kd ~ 10-15 M for chicken Avd). We previously reported engineered Avds known as antidins, which have micro- to nanomolar affinities for steroids, non-natural ligands of Avd. Here, we report the 2.8 Å X-ray structure of the sbAvd-2 (I117Y) antidin co-crystallized with progesterone. We describe the creation of new synthetic phage display libraries and report the experimental as well as computational binding analysis of progesterone-binding antidins...
2019: PloS One
Fengjiao Xu, Jinhe Chen, Cuiyan Xiao, Feng Cong, Lei Ma, Robert J Moore, Ren Huang, Pengju Guo
Resistance to aminoglycoside antibiotics is now common in pathogenic bacteria, making treatment of infections difficult. The rapid spread of resistance is mainly related to the dissemination of genes encoding aminoglycoside-modifying enzymes (AMEs). Staphylococci and enterococci are opportunistic human pathogens capable of causing a wide range of infections. Isolates from clinical cases are often found to be resistant to aminoglycosides. The aim of the present study was to develop a bead-based xTAG assay for the simultaneous detection of five prevalent aminoglycoside resistance genes in staphylococci and enterococci, including aac(6')-Ie-aph(2″)-Ia, aph(3')-IIIa, ant(4')-Ia, ant(9)-Ia, and ant(6)-Ia...
February 20, 2019: Microbial Drug Resistance: MDR: Mechanisms, Epidemiology, and Disease
Roman P Simon, Tobias Rumpf, Vaida Linkuvienė, Daumantas Matulis, Asifa Akhtar, Manfred Jung
Lysine acetyltransferases (KATs, also termed histone acetyltransferases, HATs) catalyze the acetylation of substrate lysine residues by employing the cofactor acetyl-coenzyme A (AcCoA), thereby providing a dynamic control mechanism of protein function. Because of their major involvement in cell development and homeostasis, small molecule modulators of KAT activity are urgently needed to assess their therapeutic potential and for probing their underlying biology. Recent advances in the field suggest that targeting the cofactor binding site represents a promising strategy for identifying potent and selective ligands...
February 20, 2019: Journal of Medicinal Chemistry
Roger Lafontaine Mesquita Taborda, Luiz Antônio da Silva, Patricia Puccinelli Orlandi, Flávia Serrano Batista, Renata Santos Rodrigues, Najla Benevides Matos
BACKGROUND: Enteroaggregative Escherichia coli (EAEC) is one of the main acute and chronic diarrhea causes both in children and adults, mainly in developing countries. OBJECTIVE: The aim of the present study is to characterize EAEC strains isolated from faecal samples and to identify genes potentially contributing to virulence, biofilm production and antimicrobial resistance in children admitted to a pediatric hospital in Porto Velho, Rondônia State. METHODS: The total of 1,625 E...
October 2018: Arquivos de Gastroenterologia
Mohammad Mahdi Aeinehvand, Laura Weber, Martín Jiménez, Andrea Palermo, Maria Bauer, Felix F Loeffler, Fatimah Ibrahim, Frank Breitling, Jan Korvink, Marc Madou, Dario Mager, Sergio O Martínez-Chapa
Reversible valves on centrifugal microfluidic platforms facilitate the automation of bioanalytical assays, especially of those requiring a series of steps (such as incubation) in a single reaction chamber. In this study, we present fixed elastic reversible (FER) valves and tunable elastic reversible (TER) valves that are easy to fabricate, implement and control. In the FER valve the compression of an elastic barrier/patch against a microchamber's outlet prevents the release of liquid. The valve sealing pressure was determined by adjusting the engraving depth of the valve-seat at which the elastic patch was located, this allows to set the sealing pressure during disc fabrication...
February 20, 2019: Lab on a Chip
Melissa Si Ammour, Giorgia Fedele, Caterina Morcia, Valeria Terzi, Vittorio Rossi
Quantification of colonization of grape bunch trash by Botrytis cinerea is crucial for Botrytis bunch rot (BBR) control. A previously developed qPCR method was adapted to quantify B. cinerea DNA in grape bunch trash, and a colonization coefficient (CC) was calculated as the ratio between the DNA concentrations of B. cinerea and of Vitis vinifera. CC values increased linearly with the number of conidia of B. cinerea or the quantity of mycelium of B. cinerea added to the bunch trash increased. CC values also increased linearly in bunch trash samples containing increasing percentages of B...
February 20, 2019: Phytopathology
Olga Bondareva, Roman Tsaryk, Vesna Bojovic, Maria Odenthal-Schnittler, Arndt F Siekmann, Hans-J Schnittler
AIM: Oscillatory shear stress (OSS) is an atheroprone hemodynamic force that occurs in areas of vessel irregularities and is implicated in the pathogenesis of atherosclerosis. Changes in signaling and transcriptional program in response to OSS have been vigorously studied, however, the underlying changes in the chromatin landscape controlling transcription remain to be elucidated. Here, we investigated the changes in the regulatory element (RE) landscape of endothelial cells under atheroprone OSS conditions in an in vitro model...
February 20, 2019: Cardiovascular Research
Xi He, Ping Chen, Ji Zhang, Tian-Ying Luo, Hai-Jiao Wang, Yan-Hong Liu, Xiao-Qi Yu
Carbon dots have attracted rapidly growing interest in recent years. In this report, we prepared two cationic polymer-derived carbon dots (Taea-CD and Cyclen-CD, collectively called C-dots) via a hydrothermal method. Transmission electron microscopy (TEM) results show that the C-dots were sphere-like and the size distribution was 1.8 ± 0.4 nm for Taea-CD and 5.4 ± 2 nm for Cyclen-CD. The C-dots emitted bright blue fluorescence under UV light (365 nm). Confocal laser scanning microscopy (CLSM) assay indicates that the C-dots-mediated transfection process could be detected in real time, and their tunable fluorescence emission under different wavelengths could satisfy varying requirements...
February 20, 2019: Biomaterials Science
Li Yingchun, Wang Huihan, Zhang Rong, Zhang Guojun, Yang Ying, Liu Zhuogang
BACKGROUND Accumulating evidence suggests that plant-derived molecules may prove extremely beneficial in the development of chemotherapy for deadly cancer types. Multiple myeloma is a rare and incurable type of cancers. Very little research has been directed towards the development of chemotherapy for the management of multiple myeloma. Here, the anticancer effects of a plant-derived triterpenoid, Asiaticoside, were examined against the drug-resistant myeloma cell line KM3/BTZ. MATERIAL AND METHODS Cell viability was determined by CCK-8 assay and autophagy was checked by transmission electron microscopy...
February 20, 2019: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
Yudi Xiong, Tianqi Li, Ganiou Assani, Huan Ling, Qian Zhou, Yangyang Zeng, Fuxiang Zhou, Yunfeng Zhou
Cervical cancer remains one of the main factors leading to tumor-related death worldwide. Many strategies of cancer treatment such as chemotherapy are developed and used nowadays. However, for the cancer chemotherapy resistance, reduction of the limitation of cancer chemotherapy efficacy is one of the aims of several oncology teams. Moreover, the cyclin-dependent kinase 4/6-cyclin D-retinoblastoma protein-E2F pathway is an important mechanism for cell cycle control and its dysregulation is one of the key factors for cancers development including cervical cancer...
February 18, 2019: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Piyoosh Sharma, Avanish Tripathi, Prabhash Nath Tripathi, Santosh Kumar Prajapati, Ankit Seth, Manish Kumar Tripathi, Pavan Srivastava, Vinod Tiwari, Sairam Krishnamurthy, Sushant Kumar Shrivastava
The multitarget-directed strategy offers an effective and promising paradigm to treat the complex neurodegenerative disorder, such as Alzheimer's disease (AD). Herein, a series of N-benzylpiperidine analogs (17-31 and 32-46) were designed and synthesized as multi-functional inhibitors of acetylcholinesterase (AChE) and β-secretase-1 (BACE-1) with moderate to excellent inhibitory activities. Among the tested inhibitors, 25, 26, 40, and 41 presented the most significant and balanced inhibition against both the targets...
February 14, 2019: European Journal of Medicinal Chemistry
Hugues Prevet, Martin Moune, Abdalkarim Tanina, Christian Kemmer, Adrien Herledan, Rosangela Frita, Alexandre Wohlkönig, Marilyne Bourotte, Baptiste Villemagne, Florence Leroux, Marc Gitzinger, Alain R Baulard, Benoit Déprez, René Wintjens, Nicolas Willand, Marion Flipo
Tuberculosis (TB) caused by the pathogen Mycobacterium tuberculosis, represents one of the most challenging threat to public health worldwide, and with the increasing resistance to approved TB drugs, it is needed to develop new strategies to address this issue. Ethionamide is one of the most widely used drugs for the treatment of multidrug-resistant TB. It is a prodrug that requires activation by mycobacterial monooxygenases to inhibit the enoyl-ACP reductase InhA, which is involved in mycolic acid biosynthesis...
February 10, 2019: European Journal of Medicinal Chemistry
Ivan D Tarandovskiy, Paul W Buehler, Fazoil I Ataullakhanov, Elena Karnaukhova
Package inserts for C1-esterase inhibitor (C1INH) products include warnings for an elevated risk of possible thrombosis in certain individuals, referring to thromboembolic events (TEEs) that were reported to occur after C1INH infusions. However, the mechanism(s) that could explain possible development of TEEs due to C1INH remains unknown. In this work, we evaluated plausible impact of C1INH on the protein C (PC) anticoagulant system. We performed thrombin generation (TG) assays (TGA) and analyzed spatial fibrin clot propagation using thrombodynamics in plasma of individual donors after the addition of thrombomodulin (TM) and C1INH...
February 12, 2019: Thrombosis Research
Christelle Zaghrini, Barbara Seitz-Polski, Joana Justino, Guillaume Dolla, Christine Payré, Noémie Jourde-Chiche, Anne-Els Van de Logt, Caroline Booth, Emma Rigby, Jennie Lonnbro-Widgren, Jenny Nystrom, Christophe Mariat, Zhao Cui, Jack F M Wetzels, GianMarco Ghiggeri, Laurence H Beck, Pierre Ronco, Hanna Debiec, Gérard Lambeau
Autoantibodies against phospholipase A2 receptor 1 (PLA2R1) and thrombospondin type 1 domain-containing 7A (THSD7A) are emerging as biomarkers to classify membranous nephropathy (MN) and to predict outcome or response to treatment. Anti-THSD7A autoantibodies are detected by Western blot and indirect immunofluorescence test (IIFT). Here, we developed a sensitive enzyme-linked immunosorbent assay (ELISA) optimized for quantitative detection of anti-THSD7A autoantibodies. Among 1012 biopsy-proven MN patients from 6 cohorts, 28 THSD7A-positive patients were identified by ELISA, indicating a prevalence of 2...
March 2019: Kidney International
Taylor M Nichols, Nolan W Kennedy, Danielle Tullman-Ercek
Metabolic engineers seek to produce high-value products from inexpensive starting materials in a sustainable and cost-effective manner by using microbes as cellular factories. However, pathway development and optimization can be arduous tasks, complicated by pathway bottlenecks and toxicity. Pathway organization has emerged as a potential solution to these issues, and the use of protein- or DNA-based scaffolds has successfully increased the production of several industrially relevant compounds. These efforts demonstrate the usefulness of pathway colocalization and spatial organization for metabolic engineering applications...
2019: Methods in Enzymology
Kavitha Siva, Fredrik Ek, Jun Chen, Abdul Ghani Alattar, Kristmundur Sigmundsson, Roger Olsson, Marcin Wlodarski, Thomas Lundbäck, Johan Flygare
Diamond-Blackfan anemia (DBA) is a bone marrow failure syndrome caused by mutations in ribosomal protein genes. Pathogenic mechanisms are poorly understood but involve severely reduced proliferation of erythroid precursors. Because current DBA therapies are ineffective and associated with severe side effects, disease-specific therapies are urgently needed. We hypothesized that druggable molecular pathways underlying the defect can be revealed through phenotypic small-molecule screens. Accordingly, a screening assay was developed using c-kit+ fetal liver erythroid progenitors from a doxycycline-inducible DBA mouse model...
March 2019: SLAS Discovery
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