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Reverse fusion

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https://www.readbyqxmd.com/read/28942999/optimized-purification-of-a-fusion-protein-by-reversed-phase-high-performance-liquid-chromatography-informed-by-the-linear-solvent-strength-model
#1
Isaac B Falconer, Colin T Mant, C James McKnight, Liliya Vugmeyster, Robert Hodges
Fusion protein systems are commonly used for expression of small proteins and peptides. An important criterion for a fusion protein system to be useful is the ability to separate the protein of interest from the tag. Additionally, because no protease cleaves fusion proteins with 100% efficiency, the ability to separate the desired peptide from any remaining uncleaved protein is also necessary. This is likely to be the more difficult task as at least a portion of the sequence of the fusion protein is identical to that of the protein of interest...
September 13, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28939434/inhibition-of-drp1-mediated-mitochondrial-fission-improves-mitochondrial-dynamics-and-bioenergetics-stimulating-neurogenesis-in-hippocampal-progenitor-cells-from-a-down-syndrome-mouse-model
#2
Daniela Valenti, Leonardo Rossi, Domenico Marzulli, Francesco Bellomo, Domenico De Rasmo, Anna Signorile, Rosa Anna Vacca
Functional and structural damages to mitochondria have been critically associated with the pathogenesis of Down syndrome (DS), a human multifactorial disease caused by trisomy of chromosome 21 and associated with neurodevelopmental delay, intellectual disability and early neurodegeneration. Recently, we demonstrated in neural progenitor cells (NPCs) isolated from the hippocampus of Ts65Dn mice -a widely used model of DS - a severe impairment of mitochondrial bioenergetics and biogenesis and reduced NPC proliferation...
September 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28934472/controlled-re-activation-of-epigenetically-silenced-tet-promoter-driven-transgene-expression-by-targeted-demethylation
#3
Natascha Gödecke, Lisha Zha, Shawal Spencer, Sara Behme, Pamela Riemer, Michael Rehli, Hansjörg Hauser, Dagmar Wirth
Faithful expression of transgenes in cell cultures and mice is often challenged by locus dependent epigenetic silencing. We investigated silencing of Tet-controlled expression cassettes within the mouse ROSA26 locus. We observed pronounced DNA methylation of the Tet promoter concomitant with loss of expression in mES cells as well as in differentiated cells and transgenic animals. Strikingly, the ROSA26 promoter remains active and methylation free indicating that this silencing mechanism specifically affects the transgene, but does not spread to the host's chromosomal neighborhood...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28930199/reduced-fluorescent-protein-switching-fatigue-by-binding-induced-emissive-state-stabilization
#4
Thijs Roebroek, Sam Duwé, Wim Vandenberg, Peter Dedecker
Reversibly switchable fluorescent proteins (RSFPs) enable advanced fluorescence imaging, though the performance of this imaging crucially depends on the properties of the labels. We report on the use of an existing small binding peptide, named Enhancer, to modulate the spectroscopic properties of the recently developed rsGreen series of RSFPs. Fusion constructs of Enhancer with rsGreen1 and rsGreenF revealed an increased molecular brightness and pH stability, although expression in living E. coli or HeLa cells resulted in a decrease of the overall emission...
September 20, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28929274/minimally-invasive-posterior-cervical-foraminotomy-for-treatment-of-radiculopathy-an-effective-time-tested-and-cost-efficient-motion-preservation-technique
#5
REVIEW
L Papavero, R Kothe
OBJECTIVE: To relieve foraminal root impingement due to lateral soft disc fragments, bony spurs, or other rarer causes. INDICATIONS: Soft disc fragment whose bulk is >2/3 lateral to the lateral border of the thecal sac. Intraforaminal dorsal bony narrowing of the root canal. Intraforaminal synovial cyst, extra/intradural tumor. CONTRAINDICATIONS: Paramedian and median soft/hard disc protrusions. Kyphosis of the index level. SURGICAL TECHNIQUE: Patient prone in reverse Trendelenburg position with the head fixed in a Mayfield clamp...
September 19, 2017: Operative Orthopädie und Traumatologie
https://www.readbyqxmd.com/read/28926421/a-novel-hydroxyphenyl-hydrazone-derivate-ycl0426-inhibits-cancer-cell-proliferation-through-sequestering-iron
#6
Feifei Li, Long Long, Junhai Xiao, Chen Wang, Wei Li, Song Li, Changqi Zhao, Lili Wang
Cancer cells have an increased requirement for iron than normal cells, and iron chelators are under active consideration for cancer treatment. The metal-sequestering potential and antiproliferative mechanisms of a novel hydroxyphenyl hydrazone derivate YCL0426 were investigated here. Antiproliferative activity of YCL0426 was detected by MTT assay. The iron-sequestering potential was evaluated by ferrozine-Fe(II) sequestering assay and Fe(II) titration assay. Cell-cycle-arresting profile was checked by flow cytometry and the DNA synthesis status was evaluated by BrdU incorporation assay...
September 18, 2017: Anti-cancer Drugs
https://www.readbyqxmd.com/read/28917636/parallel-approach-on-sorting-of-genes-in-search-of-optimal-solution
#7
Pranav Kumar, G Sahoo
An important tool for comparing genome analysis is the rearrangement event that can transform one given genome into other. For finding minimum sequence of fission and fusion, we have proposed here an algorithm and have shown a transformation example for converting the source genome into the target genome. The proposed algorithm comprises of circular sequence i.e. "cycle graph" in place of mapping. The main concept of algorithm is based on optimal result of permutation. These sorting processes are performed in constant running time by showing permutation in the form of cycle...
September 13, 2017: Genomics
https://www.readbyqxmd.com/read/28916764/rapid-and-reversible-epigenome-editing-by-endogenous-chromatin-regulators
#8
Simon M G Braun, Jacob G Kirkland, Emma J Chory, Dylan Husmann, Joseph P Calarco, Gerald R Crabtree
Understanding the causal link between epigenetic marks and gene regulation remains a central question in chromatin biology. To edit the epigenome we developed the FIRE-Cas9 system for rapid and reversible recruitment of endogenous chromatin regulators to specific genomic loci. We enhanced the dCas9-MS2 anchor for genome targeting with Fkbp/Frb dimerizing fusion proteins to allow chemical-induced proximity of a desired chromatin regulator. We find that mSWI/SNF (BAF) complex recruitment is sufficient to oppose Polycomb within minutes, leading to activation of bivalent gene transcription in mouse embryonic stem cells...
September 15, 2017: Nature Communications
https://www.readbyqxmd.com/read/28914467/cloning-expression-pattern-analysis-and-subcellular-localization-of-capra-hircus-scd1-gene-with-production-of-transgenic-mice
#9
Qisheng Zuo, Kai Jin, Jiuzhou Song, Yani Zhang, Bichun Li
This study aimed to clone the Stearoyl-CoA desaturase 1 (SCD1) gene derived from Xuhuai goat (Capra hircus), and analyze the sub-cellular localization in cells and tissues. The cDNA was cloned by reverse transcription polymerase chain reaction (RT-PCR). pEGFP-SCD1 vector was constructed to detect sub-cellular localization and tissue distribution. pEGFP-SCD1 was transfected into NIH-3T3 cells using polyethylene imine (PEI) and observed under fluorescence inversion microscope system 48 h after transfection. The expression level of SCD1 was detected by RT-PCR...
September 15, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28900483/sulforaphane-induced-apoptosis-via-promotion-of-mitochondrial-fusion-and-erk1-2-mediated-26s-proteasome-degradation-of-novel-pro-survival-bim-and-upregulation-of-bax-in-human-non-small-cell-lung-cancer-cells
#10
Yang Geng, Yan Zhou, Sai Wu, Yabin Hu, Kai Lin, Yalin Wang, Zhongnan Zheng, Wei Wu
Previous studies in our laboratory showed that sulforaphane (SFN) induced apoptosis by sustained activation of extracellular regulated protein kinases 1/2 (ERK1/2). However, the underlying mechanisms associated with SFN-induced apoptosis and downstream cascades which are modulated by ERK1/2 were not elucidated. Herein we demonstrated for the first time that alteration of mitochondrial dynamics contributed to SFN-induced apoptosis in human non-small cell lung cancer (NSCLC) cells. Reports showed that protein Bim not only induced apoptosis but also promoted proliferation under certain circumstances...
2017: Journal of Cancer
https://www.readbyqxmd.com/read/28891599/expression-in-escherichia-coli-of-fusion-protein-comprising-%C3%AE-conotoxin-txib-and-preservation-of-selectivity-to-nicotinic-acetylcholine-receptors-in-the-purified-product
#11
Jinpeng Yu, Xiaopeng Zhu, Yang Yang, Sulan Luo, Dongting Zhangsun
Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels, which are widely distributed in the central and peripheral nervous system. The α6β2* nAChR is an important subtype, which is closely associated with nicotine addiction and movement disorders etc. α-conotoxin TxIB with 16-amino acid residues specifically targets α6β2* nAChR with no obvious effect on other nAChR subtypes. However, chemical synthesis of TxIB is expensive, and the quantity of native TxIB extracted from cone snail is limited...
September 11, 2017: Chemical Biology & Drug Design
https://www.readbyqxmd.com/read/28890970/dynamic-protein-self-assembly-driven-by-host-guest-chemistry-and-the-folding-unfolding-feature-of-a-mutually-exclusive-protein
#12
Ruidi Wang, Shanpeng Qiao, Linlu Zhao, Chunxi Hou, Xiumei Li, Yao Liu, Quan Luo, Jiayun Xu, Hongbin Li, Junqiu Liu
A novel exploration utilizing a well-designed fusion protein containing a redox stimuli-responsive domain was developed to construct dynamic protein self-assemblies induced by cucurbit[8]uril-based supramolecular interactions. The reversible interconversion of the morphology of the assemblies between nanowires and nanorings was regulated precisely by redox conditions.
September 21, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28885344/surgical-treatment-for-lumbar-hyperlordosis-after-resection-of-a-spinal-lipoma-associated-with-spina-bifida-a-case-report
#13
Tatsuya Sato, Ikuho Yonezawa, Shingo Onda, Kei Yoshikawa, Hiromitsu Takano, Yukitoshi Shimamura, Takatoshi Okuda, Kazuo Kaneko
RATIONALE: A hyperlordosis deformity of the lumbar spine is relatively rare, and surgical treatment has not been comprehensively addressed. In this case report, we describe the clinical presentation, surgical treatment, and medium-term follow-up of a patient presenting with a progressive lumbar hyperlordosis deformity after resection of a spinal lipoma associated with spina bifida. PATIENT CONCERNS: The patient was a 20-year-old woman presenting with a progressive hyperlordosis deformity of the lumbar spine associated with significant back pain (visual analog pain score of 89/100 mm), but with no neurological symptoms...
September 2017: Medicine (Baltimore)
https://www.readbyqxmd.com/read/28879687/recombinant-synthesis-of-hybrid-lipid-peptide-polymer-fusions-that-self-assemble-and-encapsulate-hydrophobic-drugs
#14
Kelli M Luginbuhl, Davoud Mozhdehi, Michael Dzuricky, Parisa Yousefpour, Fred C Huang, Nicholas R Mayne, Kristen Buehne, Ashutosh Chilkoti
Inspired by biohybrid molecules that are synthesized in Nature through post-translational modification (PTM), we have exploited a eukaryotic PTM to recombinantly synthesize lipid-polypeptide hybrid materials. By co-expressing yeast N-myristoyltransferase with an elastin-like polypeptide (ELP) fused to a short recognition sequence in E. coli, we show robust and high yield modification of the ELP with myristic acid. The ELP's reversible phase behavior is retained upon myristoylation and can be tuned to span a 30-60 °C...
September 6, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28878349/peptide-mediated-delivery-of-donor-mitochondria-improves-mitochondrial-function-and-cell-viability-in-human-cybrid-cells-with-the-melas-a3243g-mutation
#15
Jui-Chih Chang, Fredrik Hoel, Ko-Hung Liu, Yau-Huei Wei, Fu-Chou Cheng, Shou-Jen Kuo, Karl Johan Tronstad, Chin-San Liu
The cell penetrating peptide, Pep-1, has been shown to facilitate cellular uptake of foreign mitochondria but further research is required to evaluate the use of Pep-1-mediated mitochondrial delivery (PMD) in treating mitochondrial defects. Presently, we sought to determine whether mitochondrial transplantation rescue mitochondrial function in a cybrid cell model of mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) disease. Following PMD, recipient cells had internalized donor mitochondria after 1 h, and expressed higher levels of normal mitochondrial DNA, particularly at the end of the treatment and 11 days later...
September 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28877639/novel-solidified-reverse-micellar-solution-based-mucoadhesive-nano-lipid-gels-encapsulating-miconazole-nitrate-loaded-nanoparticles-for-improved-treatment-of-oropharyngeal-candidiasis
#16
Franklin Chimaobi Kenechukwu, Anthony Amaechi Attama, Emmanuel Chinedum Ibezim
OBJECTIVE: To develop and evaluate solidified-reverse-micellar-solution (SRMS)-based oromucosal nano lipid gels for improved localised delivery of miconazole nitrate (MN). METHODS: Phospholipon(®) 90G and Softisan(®) 154 (3:7) were used to prepare SRMS by fusion. Solid lipid nanoparticles (SLNs, 0.25-1.0% w/w MN) formulated with the SRMS by high shear homogenisation were employed to prepare mucoadhesive nano lipid gels. Physicochemical characterisation, drug release in simulated salivary fluid (SSF) (pH 6...
September 6, 2017: Journal of Microencapsulation
https://www.readbyqxmd.com/read/28877060/clinicopathologic-diversity-of-undifferentiated-sarcoma-with-bcor-ccnb3-fusion-analysis-of-11-cases-with-a-reappraisal-of-the-utility-of-immunohistochemistry-for-bcor-and-ccnb3
#17
Atsuji Matsuyama, Eisuke Shiba, Yoshihisa Umekita, Kanae Nosaka, Takihiro Kamio, Hiroyuki Yanai, Chika Miyasaka, Reiko Watanabe, Ichiro Ito, Tomoko Tamaki, Shinichi Hayashi, Masanori Hisaoka
Undifferentiated sarcoma harboring the BCOR-CCNB3 fusion is characterized by its predilection to affect skeletons of adolescent males, cellular small round/spindle cell morphology, and CCNB3 immunoreactivity. We analyzed 11 cases of BCOR-CCNB3 sarcoma, 10 of which were identified in a reverse transcription-polymerase chain reaction-based screen of 85 patient samples recorded in our database as unclassified small round or spindle cell sarcomas. BCOR rearrangements were confirmed by fluorescence in situ hybridization in 8 tumors...
September 4, 2017: American Journal of Surgical Pathology
https://www.readbyqxmd.com/read/28870615/luspatercept-for-the-treatment-of-anaemia-in-patients-with-lower-risk-myelodysplastic-syndromes-pace-mds-a-multicentre-open-label-phase-2-dose-finding-study-with-long-term-extension-study
#18
Uwe Platzbecker, Ulrich Germing, Katharina S Götze, Philipp Kiewe, Karin Mayer, Jörg Chromik, Markus Radsak, Thomas Wolff, Xiaosha Zhang, Abderrahmane Laadem, Matthew L Sherman, Kenneth M Attie, Aristoteles Giagounidis
BACKGROUND: Myelodysplastic syndromes are characterised by ineffective erythropoiesis. Luspatercept (ACE-536) is a novel fusion protein that blocks transforming growth factor beta (TGF β) superfamily inhibitors of erythropoiesis, giving rise to a promising new investigative therapy. We aimed to assess the safety and efficacy of luspatercept in patients with anaemia due to lower-risk myelodysplastic syndromes. METHODS: In this phase 2, multicentre, open-label, dose-finding study (PACE-MDS), with long-term extension, eligible patients were aged 18 years or older, had International Prognostic Scoring System-defined low or intermediate 1 risk myelodysplastic syndromes or non-proliferative chronic myelomonocytic leukaemia (white blood cell count <13 000/μL), and had anaemia with or without red blood cell transfusion support...
September 1, 2017: Lancet Oncology
https://www.readbyqxmd.com/read/28869107/molecular-characterization-of-a-series-of-solitary-fibrous-tumors-including-immunohistochemical-expression-of-stat6-and-natb2-stat6-fusion-transcripts-using-reverse-transcriptase-rt-polymerase-chain-reaction-pcr-technique-an-indian-experience
#19
Bharat Rekhi, Omshree Shetty, Parul Tripathi, Prachi Bapat, Mukta Ramadwar, Jyoti Bajpai, Ajay Puri
A solitary fibrous tumor (SFT) is characterized by a diverse clinicopathologic spectrum. Recent studies have unraveled STAT6 as a useful diagnostic immunohistochemical (IHC) marker for a SFT and NAB2-STAT6 as its specific gene fusion transcript. Thirty-three SFTs were tested for STAT6 immunostaining by polymer detection technique. STAT6 immunoexpression was further graded, based on intensity (mild, moderate and strong) and percentage of immunopositive tumor cells, ranging from 1 to 25%(1+); 26-50%(2+); 51-75%(3+) and in more than 75%(4+) tumor nuclei...
August 26, 2017: Pathology, Research and Practice
https://www.readbyqxmd.com/read/28867626/enhanced-cytopathic-effect-of-japanese-encephalitis-virus-strain-sa14-14-2-probable-association-of-mutation-in-amino-acid-of-its-envelope-protein
#20
Barnali Nath, Ashutosh Gupta, Siraj A Khan, Sachin Kumar
Japanese encephalitis (JE) is a mosquito-borne viral disease. It is a global public health concern since it causes an acute encephalitis syndrome (AES). A large number of JE/AES cases are reported to occur in areas with established or developing JE vaccination program. Partial vaccine coverage and emergence of new variants of Japanese encephalitis virus (JEV) might be playing an important role. The envelope protein (E) of JEV is a major antigenic determinant and responsible for immunogenic responses as well as membrane fusion and virion assembly...
September 1, 2017: Microbial Pathogenesis
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