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Tiezheng Li, Chao Li, David N Quan, William E Bentley, Lai-Xi Wang
Chemoenzymatic glycan remodeling of antibodies using an endoglycosidase and its mutant is emerging as an attractive approach for producing homogeneous antibody glycoforms. We report in this paper a site-specific covalent immobilization of the endoglycosidases (Endo-S2 and its glycosynthase mutant D184M) using a recombinant microbial transglutaminase (MTG) and evaluation of the immobilized enzymes in deglycosylation and glycosylation of a therapeutic antibody. The site-specific covalent immobilization was achieved by introduction of a Q-tag at the C-terminus of the recombinant enzymes followed by conjugation of the enzymes to a primary amine-containing solid support through MTG-catalyzed transglutamination...
February 15, 2018: Carbohydrate Research
Yingyi Yu, Yuanyuan Shi, Xinyi Zuo, Qi Feng, Yu Hou, Wei Tang, Yi Lu, Fan Yi, Ming Hou, Yuan Yu, Jun Peng
INTRODUCTION: Progranulin (PGRN) is emerging as a critical immune mediator involved in a variety of autoimmune disorders. However, its role in immune thrombocytopenia (ITP) remains unclear. MATERIALS AND METHODS: In this study, the enzyme-linked immunosorbent assay was used for determining the plasma levels of PGRN in ITP patients vs. healthy controls. In addition, the role of PGRN in ITP was investigated in two kinds of ITP murine models. Further, we explored whether PGRN functioned by affecting the number of T regulatory cells (Tregs) using flow cytometry...
February 17, 2018: Thrombosis Research
P J M van Kleeff, J Gao, S Mol, N Zwart, H Zhang, K W Li, A H de Boer
Potassium (K+ ) is a vital ion for many processes in the plant and fine-tuned ion channels control the K+ -fluxes across the plasma membrane. GORK is an outward-rectifying K+ -channel with important functions in stomatal closure and in root K+ -homeostasis. In this study, post-translational modification of the Arabidopsis GORK ion channel and its regulation by 14-3-3 proteins was investigated. To investigate the possible interaction between GORK and 14-3-3s an in vivo pull-down from an Arabidopsis protein extract with recombinant GORK C-terminus (GORK-C) indeed identified endogenous 14-3-3s (LAMBDA, CHI, NU) as binding partners in a phosphorylation dependent manner...
February 16, 2018: Plant Physiology and Biochemistry: PPB
Ling-Ling Zhao, Kaimin Hui, Yue Wang, Hui Wang, Qian Ren, Xin-Cang Li
Galactoside-binding lectins, also known as galectins, play crucial roles in innate immune response in invertebrates. In this study, three cDNA sequences from Hyriopsis cumingii were identified and collectively called HcGalec genes. Each of the three deduced HcGalec proteins contained a galactose-binding lectin domain or a GLECT domain. All the three HcGalec genes are mainly present in the hepatopancreas and gills, and their expression is induced by bacterial challenge. Three recombinant HcGalec proteins can bind and agglutinate (Ca2+ -dependent) various microorganisms, including Gram-positive and Gram-negative bacteria...
February 20, 2018: Fish & Shellfish Immunology
Meijun Yuan, Mingxiao Ning, Panpan Wei, Wenjing Hao, Yunting Jing, Wei Gu, Wen Wang, Qingguo Meng
Serpin families classified serine protease inhibitors regulate various physiological processes. However, there is not study on the role of serpin in immune responses against Spiroplasma eriocheiris as a novel causative pathogen in the Chinese mitten crab, Eriocheir sinensis. In our study, quantitative real-time PCR (qRT-PCR) revealed that the mRNA transcripts of Esserpin-2 were ubiquitous in every tissue, relative higher expression in hepatopancreas, gill and hemocytes, while the intestine, muscle, heart and nerve showed relative lower expression...
February 20, 2018: Fish & Shellfish Immunology
Yeon Duk Woo, Jaemoon Koh, Hye-Ryun Kang, Hye Young Kim, Doo Hyun Chung
BACKGROUND: The XCL1-XCR1 axis has been reported to play a role in immune homeostasis and inflammation. However, it is not known whether this axis has a critical function in allergic asthma. OBJECTIVE: In the present study, we explored that the iNKT cell-mediated XCL1-XCR1 axis regulated the allergic asthma. METHODS: Ovalbumin (OVA) or house dust mite (HDM)-induced asthma was developed in XCL1 or XCR1 knockout (KO) mice. RESULTS: XCL1 or XCR1 KO mice showed attenuation in airway hyperresponsiveness (AHR), numbers of CD103+ dendritic cells (DCs), and Th2 responses in the lungs compared with wild-type (WT) mice during OVA or HDM-induced asthma...
February 20, 2018: Journal of Allergy and Clinical Immunology
Kelly A Curtis, Daphne Morrison, Donna L Rudolph, Anupama Shankar, Laura S P Bloomfield, William M Switzer, S Michele Owen
Isothermal nucleic acid amplification techniques, such as reverse-transcription loop-mediated isothermal amplification (RT-LAMP), exhibit characteristics that are suitable for the development of a rapid, low-cost NAT that can be used at the POC. For demonstration of utility for global use, studies are needed to validate the performance of RT-LAMP for the detection of divergent subtypes. In this study, we designed and evaluated multiplexed HIV-1 integrase RT-LAMP primers to detect subtypes within group M, along with an RNase P positive internal processing and amplification control...
February 20, 2018: Journal of Virological Methods
Aleksi Sipola, Pekka Marttinen, Jukka Corander
Summary: The advent of genomic data from densely sampled bacterial populations has created a need for flexible simulators by which models and hypotheses can be efficiently investigated in the light of empirical observations. Bacmeta provides fast stochastic simulation of neutral evolution within a large collection of interconnected bacterial populations with completely adjustable connectivity network. Stochastic events of mutations, recombinations, insertions/deletions, migrations and microepidemics can be simulated in discrete non-overlapping generations with a Wright-Fisher model that operates on explicit sequence data of any desired genome length...
February 20, 2018: Bioinformatics
Tomoki Yoshikawa, Hikaru Fujii, Akiko Okutani, Miho Shibamura, Natsumi Omura, Kazutaka Egawa, Hirofumi Kato, Takuya Inagaki, Shizuko Harada, Souichi Yamada, Shigeru Morikawa, Masayuki Saijo
LC16m8 (m8), a highly attenuated vaccinia virus (VAC) strain, was developed as a smallpox vaccine, and its safety and immunogenicity have been confirmed. Here, we aimed to develop a system that recovers infectious m8 from a bacterial artificial chromosome (BAC) that retains the full-length viral genomic DNA (m8-BAC system). The infectious virus was successfully recovered from a VAC-BAC plasmid, named pLC16m8-BAC. Furthermore, the bacterial replicon-free virus was generated by intramolecular homologous recombination and was successfully recovered from a modified VAC-BAC plasmid, named pLC16m8...
2018: PloS One
Michael White, Andrew Freistaedter, Gwendolyn J B Jones, Emmanuel Zervos, Rachel L Roper
Pancreatic cancer is the 5th leading cause of cancer deaths, and there are no effective treatments. We developed a poxvirus platform vaccine with improved immunogenicity and inserted the mesothelin gene to create an anti-mesothelin cancer vaccine. Mesothelin expression is mostly restricted to tumors in adult mammals and thus may be a good target for cancer treatment. We show here that the modified vaccinia virus Ankara (MVA) virus expressing mesothelin and the enhanced MVA virus missing the immunosuppressive A35 gene and expressing mesothelin were both safe in mice and were able to induce IFN-gamma secreting T cells in response to mesothelin expressing tumor cells...
2018: PloS One
Azam Hemmati, Hadi Hassannia, Saeideh Milani, Reza Hadavi, Fatemeh Ghaemimanesh, Hodjattallah Rabbani
Receptor tyrosine kinase ROR1 has been introduced as an interesting prognostic cancer marker in histopathology. The aim of this study was to produce a polyclonal antibody (PAb) against recombinant human ROR1 protein to be used as a tool for investigation of ROR1 expression in human cancer tissue blocks. The extracellular part of human ROR1 recombinant protein was expressed using pET-28b(+) plasmid in Escherichia coli Bl21(DE3) host. The recombinant ROR1, as a candidate immunogen, was purified and injected to a New Zealand rabbit...
February 2018: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Joseph A Harvey, Laura S Itzhaki, Ewan Main
Harnessing and controlling self-assembly is an important step in developing proteins as novel biomaterials. With this goal, here we report the design of a general genetically programmed system that covalently concatenates multiple distinct protein domains into specific assembled arrays. It is driven by iterative intein-mediated Native Chemical Ligation (NCL) under mild native conditions. The system uses a series of initially inert recombinant protein fusions that sandwich the protein modules to be ligated between one of a number of different affinity tags and an intein protein domain...
February 23, 2018: ACS Synthetic Biology
Francisco Solano
The production of pigment in mammalian melanocytes requires the contribution of at least three melanogenic enzymes, tyrosinase and two other accessory enzymes called the tyrosinase-related proteins (Trp1 and Trp2), which regulate the type and amount of melanin. The last two proteins are paralogues to tyrosinase, and they appeared late in evolution by triplication of the tyrosinase gene. Tyrosinase is a copper-enzyme, and Trp2 is a zinc-enzyme. Trp1 has been more elusive, and the direct identification of its metal cofactor has never been achieved...
February 23, 2018: International Journal of Molecular Sciences
Suman Bishnoi, Ritudhwaj Tiwari, Sharad Gupta, Siddappa N Byrareddy, Debasis Nayak
Modern oncotherapy approaches are based on inducing controlled apoptosis in tumor cells. Although a number of apoptosis-induction approaches are available, site-specific delivery of therapeutic agents still remain the biggest hurdle in achieving the desired cancer treatment benefit. Additionally, systemic treatment-induced toxicity remains a major limiting factor in chemotherapy. To specifically address drug-accessibility and chemotherapy side effects, oncolytic virotherapy (OV) has emerged as a novel cancer treatment alternative...
February 23, 2018: Viruses
Xuelian Wu, Judy Nancy Hart, Xiaoming Wen, Liang Wang, Yi Du, Shixue Dou, Yun Hau Ng, Rose Amal, Jason Scott
It has been reported that photogenerated electrons and holes can be directed toward specific crystal facets of a semiconductor particle, which is believed to arise from differences in their surface electronic structures, suggesting that different facets can act as either photo-reduction or photo-oxidation sites. This study examines the propensity for this effect to occur in faceted, plate-like bismuth molybdate (Bi2MoO6), which is a useful photocatalyst for water oxidation. Photo-excited electrons and holes are shown to be spatially separated toward the {100} and {001}/{010} facets of Bi2MoO6, respectively, by facet-dependent photo-deposition of noble metals (Pt, Au and Ag) and metal oxides (PbO2, MnOx and CoOx)...
February 23, 2018: ACS Applied Materials & Interfaces
Michael Przybylski
Alpha-galactosidase (αGal) is a lysosomal enzyme that hydrolyses the alphagalactosyl moiety from glycosphingo-lipids. Mutations in the αGal genes lead to defect enzyme resulting in substrate accumulation and pathophysiology. The deficiency of αGal, called Fabry's Disease (FD), belongs to the lysosomal storage diseases. Effective treatment of FD has been developed by enzyme replacement therapy (ERT) by infusion of recombinant enzyme to increase enzyme levels and reduce accumulated substrate. Immuno-reactivity and IgG antibody formation are major, therapy-limiting complications of ERT...
February 23, 2018: ChemMedChem
Pavla Stojkova, Petra Spidlova, Juraj Lenco, Helena Rehulkova, Lucie Kratka, Jiri Stulik
The nucleoid-associated HU proteins are small abundant DNA-binding proteins in bacterial cell which play an important role in the initiation of DNA replication, cell division, SOS response, control of gene expression and recombination. HU proteins bind to double stranded DNA non-specifically, but they exhibit high affinity to abnormal DNA structures as four-way junctions, gaps or nicks, which are generated during DNA damage. In many pathogens HU proteins regulate expression of genes involved in metabolism and virulence...
February 23, 2018: Virulence
Alexander Gorka, Tsuyoshi Yamamoto, Jianjian Zhu, Martin Schnermann
Optical control over protein expression could provide a means to interrogate a range of biological processes. One approach has employed caged ligands of the estrogen receptor (ER) in combination with broadly used ligand-dependent Cre recombinase proteins. Existing approaches used UV or blue wavelengths, which hinders their application in tissue settings. Additionally, issues of payload diffusion can impede fine spatial control over the recombination process. Here we detail the chemical optimization a near-infrared (NIR) light-activated variant of the ER antagonist cyclofen...
February 22, 2018: Chembiochem: a European Journal of Chemical Biology
Solmaz Saboohi, Hans J Griesser, Bryan R Coad, Robert D Short, Andrew Michelmore
Historically, there have been two opposing views regarding deposition mechanisms in plasma polymerisation, radical growth and direct ion deposition, with neither being able to fully explain the chemistry of the resultant coating. Deposition rate and film chemistry are dependent on the chemistry of the plasma phase and thus the activation mechanisms of species in the plasma are critical to understanding the relative contributions of various chemical and physical routes to plasma polymer formation. In this study, we investigate the roles that hydrogen plays in activating and deactivating reactive plasma species...
February 23, 2018: Physical Chemistry Chemical Physics: PCCP
Na Liu, ChiYung Yam
As an alternative to methylammonium lead triiodide (MAPbI3 ), formamidinium lead triiodide (FAPbI3 ) perovskites have recently attracted significant attention because of their higher stability and smaller band gaps. Here, based on first-principles calculations, we investigate systematically the intrinsic defects in FAPbI3 . While methylammonium (MA)-related defects MAI and IMA in MAPbI3 have high formation energies, we found that formamidinium (FA)-related defects VFA , FAI and IFA in FAPbI3 have much lower formation energies...
February 23, 2018: Physical Chemistry Chemical Physics: PCCP
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