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https://www.readbyqxmd.com/read/28933792/a-common-haplotype-containing-functional-cacna1h-variants-is-frequently-coinherited-with-increased-tpsab1-copy-number
#1
Jonathan J Lyons, Stephanie C Stotz, Jack Chovanec, Yihui Liu, Katie L Lewis, Celeste Nelson, Thomas DiMaggio, Nina Jones, Kelly D Stone, Heejong Sung, Leslie G Biesecker, Michael A Colicos, Joshua D Milner
PurposeCaV3.2 signaling contributes to nociception, pruritus, gastrointestinal motility, anxiety, and blood pressure homeostasis. This calcium channel, encoded by CACNA1H, overlaps the human tryptase locus, wherein increased TPSAB1 copy number causes hereditary α-tryptasemia. Germ-line CACNA1H variants may contribute to the variable expressivity observed with this genetic trait.MethodsTryptase-encoding sequences at TPSAB1 and TPSB2, and TPSG1 and CACNA1H variants were genotyped in 46 families with hereditary α-tryptasemia syndrome...
September 21, 2017: Genetics in Medicine: Official Journal of the American College of Medical Genetics
https://www.readbyqxmd.com/read/28933646/visualizing-trna-dependent-mistranslation-in-human-cells
#2
Jeremy T Lant, Matthew D Berg, Daniel Hw Sze, Kyle S Hoffman, Ibukunoluwa C Akinpelu, Matthew Turk, Ilka U Heinemann, Martin L Duennwald, Christopher J Brandl, Patrick O'Donoghue
High-fidelity translation and a strictly accurate proteome were originally assumed as essential to life and cellular viability. Yet recent studies in bacteria and eukaryotic model organisms suggest that proteome-wide mistranslation can provide selective advantages and is tolerated in the cell at higher levels than previously thought (one error in 6.9 × 10(-4) in yeast) with a limited impact on phenotype. Previously, we selected a tRNA(Pro) containing a single mutation that induces mistranslation with alanine at proline codons in yeast...
September 21, 2017: RNA Biology
https://www.readbyqxmd.com/read/28928504/bioactivity-based-antioxidative-components-screening-and-evaluation-in-grape-seed-proanthocyanidin-extract
#3
Jie Gao, Shengtao Lv, Changzhi Li, Jin Tao, Chengqi Jia, Min Jiang, Yuanyuan Hou, Xiaotao Hou, Jiagang Deng, Gang Bai
Grape seed proanthocyanidin extract (GSPE), a type of functional food, possesses potent antioxidant activity. In this study, GSPE protected human embryonic kidney 293 (HEK 293) cells from H2O2-induced cell injury and oxidative stress in a dose-dependent manner. The key effective constituents that exerted the most potent antioxidative activity in GSPE were screened by using a modified ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF MS) integrated 2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) radical cation antioxidative activity analysis system...
August 2017: Journal of Food Science and Technology
https://www.readbyqxmd.com/read/28928437/as1411-aptamer-anionic-linear-globular-dendrimer-g2-iohexol-selective-nano-theranostics
#4
Pardis Mohammadzadeh, Reza Ahangari Cohan, Seyedeh Masoumeh Ghoreishi, Ahmad Bitarafan-Rajabi, Mehdi Shafiee Ardestani
Molecular theranostics is of the utmost interest for diagnosis as well as treatment of different malignancies. In the present study, anionic linear globular dendrimer G2 is employed as a suitable carrier for delivery and AS1411 aptamer is exploited as the targeting agent to carry Iohexol specifically to the human breast cancer cells (MCF-7). Dendrimer G2 was prepared and conjugation of dendrimer and aptamer was carried out thereafter. Based on the data yielded by AFM, morphology of smooth and spherical non-targeted dendrimer changed to the rough aspherical shape when it conjugated...
September 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28923388/synthesis-of-cytotoxically-active-derivatives-based-on-alkylated-2-3-seco-triterpenoids
#5
Anastasia V Konysheva, Vladimir O Nebogatikov, Irina A Tolmacheva, Maxim V Dmitriev, Victoria V Grishko
Extremely low content of biologically active triterpenoids with the fragmented or contracted ring A extractable from plants is the main disadvantage of their use in drug discovery and practical pharmacology. Development of new methods for synthesis of these compounds and their structural analogs from bioavailable triterpene precursors gives an opportunity to obtain promising agents for pharmacology with excellent yields. A new approach to synthesis of alkylated A-seco-triterpenoids, including the Beckmann fragmentation of 3-methyl-substituted allobetulin or betulinic acid methyl ester with 2-hydroxyimino group in the ring A was proposed...
September 6, 2017: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28921433/large-scale-transient-transfection-of-suspension-mammalian-cells-for-vlp-production
#6
Laura Cervera, Amine A Kamen
Large-scale transient transfection of mammalian cell suspension cultures enables the production of biological products in sufficient quantity and under stringent quality attributes to perform accelerated in vitro evaluations and has the potential to support preclinical or even clinical studies. Here we describe the methodology to produce VLPs in a 3L bioreactor, using suspension HEK 293 cells and PEIPro as a transfection reagent. Cells are grown in the bioreactor to 1 × 10(6) cells/mL and transfected with a plasmid DNA-PEI complex at a ratio of 1:2...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28919489/zinc-chloride-induced-trpa1-activation-does-not-contribute-to-toxicity-in-vitro
#7
Dirk Steinritz, Franziska Zehfuß, Bernhard Stenger, Annette Schmidt, Tanja Popp, Kai Kehe, Harald Mückter, Horst Thiermann, Thomas Gudermann
The hygroscopic zinc chloride (ZnCl2) is often used to generate smoke screens. Severe adverse pulmonary health effects have been associated with inhalation of ZnCl2 smokes. The underlying molecular toxicology is not known. Recent studies have shown that the Transient Receptor Potential Channel A1 (TRPA1) is important for sensing toxic chemicals. TRPA1 was shown to be activated by Zn(2+) which was linked to pain and inflammation. In the present study, we investigated whether TRPA1 activation contributes to ZnCl2-mediated toxicity in vitro...
September 14, 2017: Toxicology Letters
https://www.readbyqxmd.com/read/28915601/mir-148a-impairs-ras-erk-signaling-in-b-lymphocytes-by-targeting-sos-proteins
#8
Julia Alles, Nicole Ludwig, Stefanie Rheinheimer, Petra Leidinger, Friedrich A Grässer, Andreas Keller, Eckart Meese
Although microRNAs have been recognized as central cellular regulators, there is an evident lack of knowledge about their targets. Here, we analyzed potential target genes for miR-148a functioning in Ras signaling in B cells, including SOS1 and SOS2. A dual-luciferase reporter assay showed significantly decreased luciferase activity upon ectopic overexpression of miR-148a in HEK-293T cells that were co-transfected with the 3'UTR of either SOS1 or SOS2. Each of the 3'UTRs of SOS1 and SOS2 contained two binding sites for miR-148a both of which were necessary for the decreased luciferase activity...
August 22, 2017: Oncotarget
https://www.readbyqxmd.com/read/28910156/antiproliferative-effects-of-bacillus-coagulans-unique-is2-in-colon-cancer-cells
#9
Ratna Sudha Madempudi, Arunasree M Kalle
In the present study, the in vitro anticancer (antiproliferative) effects of Bacillus coagulans Unique IS2 were evaluated on human colon cancer (COLO 205), cervical cancer (HeLa), and chronic myeloid leukemia (K562) cell lines with a human embryonic kidney cell line (HEK 293T) as noncancerous control cells. The Cytotoxicity assay (MTT) clearly demonstrated a 22%, 31.7%, and 19.5% decrease in cell proliferation of COLO 205, HeLa, and K562 cells, respectively, when compared to the noncancerous HEK 293T cells...
September 14, 2017: Nutrition and Cancer
https://www.readbyqxmd.com/read/28902888/dendrimer-based-selective-autophagy-induction-rescues-%C3%AE-f508-cftr-and-inhibits-pseudomonas-aeruginosa-infection-in-cystic-fibrosis
#10
Scott Mackenzie Brockman, Manish Bodas, David Silverberg, Ajit Sharma, Neeraj Vij
BACKGROUND: Cystic Fibrosis (CF) is a genetic disorder caused by mutation(s) in the CF-transmembrane conductance regulator (Cftr) gene. The most common mutation, ΔF508, leads to accumulation of defective-CFTR protein in aggresome-bodies. Additionally, Pseudomonas aeruginosa (Pa), a common CF pathogen, exacerbates obstructive CF lung pathology. In the present study, we aimed to develop and test a novel strategy to improve the bioavailability and potentially achieve targeted drug delivery of cysteamine, a potent autophagy-inducing drug with anti-bacterial properties, by developing a dendrimer (PAMAM-DEN)-based cysteamine analogue...
2017: PloS One
https://www.readbyqxmd.com/read/28902504/synthesis-of-altrose-poly-amido-saccharides-with-%C3%AE-n-1%C3%A2-2-d-amide-linkages-a-right-handed-helical-conformation-engineered-in-at-the-monomer-level
#11
Ruiqing Xiao, Eric L Dane, Jialiu Zeng, Christopher James McKnight, Mark W Grinstaff
The design and synthesis of amide-linked saccharide oligomers and polymers, which are predisposed to fold into specific ordered secondary structures, is of significant interest. Herein, right-handed helical poly-amido-saccharides (PASs) with β-N-(1→2)-D-amide linkages are synthesized by the anionic ring-opening polymerization of an altrose β-lactam monomer (alt-lactam). The right-handed helical conformation is engineered into the polymer by pre-installing the β configuration of the lactam ring in the monomer via the stereospecific [2+2] cycloaddition of trichloroacetyl isocyanate with a D-glycal possessing a 3-benzyloxyl group oriented to the α-face of the pyranose...
September 13, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28901319/expression-and-significance-of-twist-estrogen-receptor-and-e-cadherin-in-human-breast-cancer-cells-and-tissues
#12
Ruizhi Tan, Li Wang, Jie Song, Jianchun Li, Tao He
OBJECTIVES: Breast cancer is one of the most common malignancies in women, and the tumor cells' invasion and metastasis is the main cause of death. Recent reports showed that Twist, a transcription factor, plays multiple roles in breast cancer initiation, progress, and metastasis. However, the underlying mechanisms of Twist in tumor invasion and metastasis of breast cancer still remain unclear. Here, we examined the correlation between Twist, E-cadherin, and estrogen receptor (ER) in promoting invasion and metastasis in breast cancer cells and tissues...
2017: Journal of Cancer Research and Therapeutics
https://www.readbyqxmd.com/read/28900772/clathrin-dependent-internalization-signaling-and-metabolic-processing-of-guanylyl-cyclase-natriuretic-peptide-receptor-a
#13
Naveen K Somanna, Indra Mani, Satyabha Tripathi, Kailash N Pandey
Cardiac hormones, atrial and brain natriuretic peptides (ANP and BNP), have pivotal roles in renal hemodynamics, neuroendocrine signaling, blood pressure regulation, and cardiovascular homeostasis. Binding of ANP and BNP to the guanylyl cyclase/natriuretic peptide receptor-A (GC-A/NPRA) induces rapid internalization and trafficking of the receptor via endolysosomal compartments, with concurrent generation of cGMP. However, the mechanisms of the endocytotic processes of NPRA are not well understood. The present study, using (125)I-ANP binding assay and confocal microscopy, examined the function of dynamin in the internalization of NPRA in stably transfected human embryonic kidney-293 (HEK-293) cells...
September 12, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28899725/nitric-oxide-donor-ru-terpy-bdq-no-3-induces-uncoupling-and-phosphorylation-of-endothelial-nitric-oxide-synthase-promoting-oxidant-production
#14
Simone R Potje, Zhenlong Chen, Suellen D'Arc S Oliveira, Lusiane M Bendhack, Roberto S da Silva, Marcelo G Bonini, Cristina Antoniali, Richard D Minshall
[Ru(terpy)(bdq)NO](3+) (TERPY) is a nitric oxide (NO) donor that promotes relaxation of the mesenteric artery and aorta in rats. We sought to investigate whether it acts as both an NO donor and endothelial NO synthase (eNOS) activator, as shown previously for nitroglycerin. Human umbilical vein endothelial cells (HUVECs) and human embryonic kidney 293 cells transfected with empty vector (HEK) or eNOS cDNA (HEK-eNOS) were treated with TERPY (1µM) for different lengths of time. eNOS expression, dimerization, and Ser(1177) phosphorylation, caveolin-1 (Cav-1) oligomerization, Cav-1 Tyr(14) phosphorylation were evaluated by Western blotting...
September 9, 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/28898905/taguchi-experimental-design-for-optimization-of-recombinant-human-growth-hormone-production-in-cho-cell-lines-and-comparing-its-biological-activity-with-prokaryotic-growth-hormone
#15
Zahra Sadat Aghili, Sayyed Hamid Zarkesh-Esfahani
Growth hormone deficiency results in growth retardation in children and the GH deficiency syndrome in adults and they need to receive recombinant-GH in order to rectify the GH deficiency symptoms. Mammalian cells have become the favorite system for production of recombinant proteins for clinical application compared to prokaryotic systems because of their capability for appropriate protein folding, assembly, post-translational modification and proper signal. However, production level in mammalian cells is generally low compared to prokaryotic hosts...
September 12, 2017: Drug Research
https://www.readbyqxmd.com/read/28893906/n-terminal-splicing-extensions-of-the-human-myo1c-gene-fine-tune-the-kinetics-of-the-three-full-length-myosin-ic-isoforms
#16
Lilach Zattelman, Ronit Regev, Marko Ušaj, Patrick Y A Reinke, Sven Giese, Abraham O Samson, Manuel H Taft, Dietmar J Manstein, Arnon Henn
The MYO1C gene produces three alternatively spliced isoforms, differing only in their N-terminal regions (NTRs). These isoforms, which exhibit both specific and overlapping nuclear and cytoplasmic functions, have different expression levels and nucleo-cytoplasmic partitioning. To investigate the effect of NTR extensions on the enzymatic behavior of individual isoforms, we over-expressed and purified the three full-length human isoforms from suspension-adapted HEK cells. MYO1C(C) favored the actomyosin closed state (AM(C)), MYO1C(16) populated the actomyosin open state (AM(O)) and AM(C) equally, and MYO1C(35) favored the AM(O) state...
September 11, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28892414/efficacy-of-recombinant-adenovirus-expressing-a-fusion-gene-from-gm-csf-and-epstein-barr-virus-lmp2a-in-a-mouse-tumor-model
#17
Qingjie Xue, Xiuzheng Li, Chunqing Yang, Bingyuan Ji, Yunqing Li, Yingchun Yan, Xinxin Yang, Chunmei Wang, Ting Chen
In this study, purified GM-CSF and LMP2A mRNAs were amplified by PCR. Then, the GM-CSF and LMP2A sequences were connected by the polypeptide linker (Gly4Ser)3 using gene splicing by overlap extension. The constructed fusion gene GC2A was inserted into the adenovirus vector. Then the recombinant vector was introduced into HEK 293T cells by calcium phosphate transfection to package the adenovirus. The levels of antibodies against the GM-CSF and LMP2Afusion proteins were measured by ELISA, and the CTL activity of the mouse splenic lymphocytes was determined by lactate dehydrogenase (LDH) release assay...
September 11, 2017: Human Vaccines & Immunotherapeutics
https://www.readbyqxmd.com/read/28888009/synthesis-and-evaluation-of-the-cytotoxic-and-anti-proliferative-properties-of-zno-quantum-dots-against-mcf-7-and-mda-mb-231-human-breast-cancer-cells
#18
Roshini A, Srikanth Jagadeesan, Young-Jae Cho, Jong-Hwan Lim, Kyung Hyun Choi
Current trends in therapeutic research are the application of nanomaterial carriers for cancer therapy. One such molecule, ZnO, originally used in diagnosis and as a drug carrier, is gaining importance for its biological properties. Here, we report for the first time, the scope of ZnO QDs for enhanced cytotoxicity against MCF-7 and metastatic MDA-MB-231 human breast cancer cells. Unlike other ZnO nanostructures, ZnO QDs are dispersed and small sized (8-10nm) which is believed to greatly increase the cellular uptake...
December 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
https://www.readbyqxmd.com/read/28887969/viability-of-hek-293-cells-on-poly-%C3%AE-hydroxybutyrate-phb-biosynthesized-from-a-mutant-azotobacter-vinelandii-strain-cast-film-and-electrospun-scaffolds
#19
Angel Romo-Uribe, Angelica Meneses-Acosta, Maraolina Domínguez-Díaz
Sterilization, cytotoxicity and cell viability are essential properties defining a material for medical applications and these characteristics were investigated for poly(β-hydroxybutyrate) (PHB) of 230kDa obtained by bacterial synthesis from a mutant strain of Azotobacter vinelandii. Cell viability was investigated for two types of PHB scaffolds, solution cast films and non-woven electrospun fibrous membranes, and the efficiency was compared against a culture dish. The biosynthesized PHB was sterilized by ultraviolet radiation and autoclave, it was found that the thermal properties and intrinsic viscosity remained unchanged indicating that the sterilization methods did not degrade the polymer...
December 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
https://www.readbyqxmd.com/read/28887303/dynamic-nuclear-polarization-facilitates-monitoring-of-pyruvate-metabolism-in-trypanosoma-brucei
#20
You Zhuo, Ciro D Cordeiro, S Khan Hekmatyar, Roberto Docampo, James H Prestegard
Dynamic nuclear polarization (DNP) provides sensitivity improvements that make NMR a viable method for following metabolic conversions in real time. There are now many in vivo applications to animal systems and even to diagnosis of human disease. However, application to microbial systems is rare. Here we demonstrate its application to the pathogenic protozoan, Trypanosoma brucei, using hyperpolarized (13)C1- pyruvate as a substrate and compare the parasite metabolism to that of commonly cultured mammalian cell lines, HEK-293 and Hep-G2...
September 8, 2017: Journal of Biological Chemistry
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