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https://www.readbyqxmd.com/read/29138463/talen-mediated-shift-of-mitochondrial-dna-heteroplasmy-in-melas-ipscs-with-m-13513g-a-mutation
#1
Naoki Yahata, Yuji Matsumoto, Minoru Omi, Naoki Yamamoto, Ryuji Hata
Induced pluripotent stem cells (iPSCs) are suitable for studying mitochondrial diseases caused by mitochondrial DNA (mtDNA) mutations. Here, we generated iPSCs from a patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) with the m.13513G>A mutation. The patient's dermal fibroblasts were reprogrammed, and we established two iPSC clones with and without mutant mtDNA. Furthermore, we tried to decrease mutant mtDNA level in iPSCs using transcription activator-like effector nucleases (TALENs)...
November 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29132331/adaptation-of-the-goldenbraid-modular-cloning-system-and-creation-of-a-toolkit-for-the-expression-of-heterologous-proteins-in-yeast-mitochondria
#2
Ana Pérez-González, Ryan Kniewel, Marcel Veldhuizen, Hemant K Verma, Mónica Navarro-Rodríguez, Luis M Rubio, Elena Caro
BACKGROUND: There is a need for the development of synthetic biology methods and tools to facilitate rapid and efficient engineering of yeast that accommodates the needs of specific biotechnology projects. In particular, the manipulation of the mitochondrial proteome has interesting potential applications due to its compartmentalized nature. One of these advantages resides in the fact that metalation occurs after protein import into mitochondria, which contains pools of iron, zinc, copper and manganese ions that can be utilized in recombinant metalloprotein metalation reactions...
November 13, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29128917/heterologous-expression-of-mitochondrial-nicotinamide-adenine-dinucleotide-transporter-ndt1-from-aspergillus-fumigatus-rescues-impaired-growth-in-%C3%AE-ndt1%C3%AE-ndt2-saccharomyces-cerevisiae-strain
#3
Laís de Lourdes de Lima Balico, Emerson de Souza Santos, Silveli Suzuki-Hatano, Lucas Oliveira Sousa, Ana Elisa Caleiro Seixas Azzolini, Yara Maria Lucisano-Valim, Taisa Magnani Dinamarco, Vinicius Kannen, Sérgio Akira Uyemura
Our understanding of nicotinamide adenine dinucleotide mitochondrial transporter 1 (Ndt1A) in Aspergillus fumigatus remains poor. Thus, we investigated whether Ndt1A could alter fungi survival. To this end, we engineered the expression of an Ndt1A-encoding region in a Δndt1Δndt2 yeast strain. The resulting cloned Ndt1A protein promoted the mitochondrial uptake of nicotinamide adenine dinucleotide (NAD(+)), generating a large mitochondrial membrane potential. The NAD(+) carrier utilized the electrochemical proton gradient to drive NAD(+) entrance into mitochondria when the mitochondrial membrane potential was sustained by succinate...
November 11, 2017: Journal of Bioenergetics and Biomembranes
https://www.readbyqxmd.com/read/29127121/pkc-epsilon-is-a-key-regulator-of-mitochondrial-redox-homeostasis-in-acute-myeloid-leukemia
#4
Daniela Di Marcantonio, Esteban Martinez, Simone Sidoli, Jessica Vadaketh, Margaret Nieborowska-Skorska, Anushk Gupta, Jacob Michael Meadows, Francesca Ferraro, Elena Masselli, Grant A Challen, Michael D Milsom, Claudia Scholl, Stefan Froehling, Siddharth Balachandran, Tomasz Skorski, Benjamin Garcia, Prisco Mirandola, Giuliana Gobbi, Ramiro Garzon, Marco Vitale, Stephen M Sykes
PURPOSE: The intracellular redox environment of acute myeloid leukemia (AML) cells is often highly oxidized compared to healthy hematopoietic progenitors and this is purported to contribute to disease pathogenesis. However, the redox regulators that allow AML cell survival in this oxidized environment remain largely unknown. EXPERIMENTAL DESIGN AND RESULTS: We show that RNA interference-mediated inhibition of the serine/threonine kinase PKC-epsilon (PKCe) reduces cell survival in a diverse panel of patient-derived AML samples and significantly delays disease onset in a genetically engineered mouse model (GEMM) of AML driven by MLL-AF9...
November 10, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/29121949/quantifying-engineered-nanomaterial-toxicity-comparison-of-common-cytotoxicity-and-gene-expression-measurements
#5
Donald H Atha, Amber Nagy, Andrea Steinbrück, Allison M Dennis, Jennifer A Hollingsworth, Varsha Dua, Rashi Iyer, Bryant C Nelson
BACKGROUND: When evaluating the toxicity of engineered nanomaterials (ENMS) it is important to use multiple bioassays based on different mechanisms of action. In this regard we evaluated the use of gene expression and common cytotoxicity measurements using as test materials, two selected nanoparticles with known differences in toxicity, 5 nm mercaptoundecanoic acid (MUA)-capped InP and CdSe quantum dots (QDs). We tested the effects of these QDs at concentrations ranging from 0.5 to 160 µg/mL on cultured normal human bronchial epithelial (NHBE) cells using four common cytotoxicity assays: the dichlorofluorescein assay for reactive oxygen species (ROS), the lactate dehydrogenase assay for membrane viability (LDH), the mitochondrial dehydrogenase assay for mitochondrial function, and the Comet assay for DNA strand breaks...
November 9, 2017: Journal of Nanobiotechnology
https://www.readbyqxmd.com/read/29107503/abrogating-mitochondrial-dynamics-in-mouse-hearts-accelerates-mitochondrial-senescence
#6
Moshi Song, Antonietta Franco, Julie A Fleischer, Lihong Zhang, Gerald W Dorn
Mitochondrial fusion and fission are critical to heart health; genetically interrupting either is rapidly lethal. To understand whether it is loss of, or the imbalance between, fusion and fission that underlies observed cardiac phenotypes, we engineered mice in which Mfn-mediated fusion and Drp1-mediated fission could be concomitantly abolished. Compared to fusion-defective Mfn1/Mfn2 cardiac knockout or fission-defective Drp1 cardiac knockout mice, Mfn1/Mfn2/Drp1 cardiac triple-knockout mice survived longer and manifested a unique pathological form of cardiac hypertrophy...
October 21, 2017: Cell Metabolism
https://www.readbyqxmd.com/read/29097312/insights-on-the-conformational-dynamics-of-human-frataxin-through-modifications-of-loop-1
#7
Martín E Noguera, Martín Aran, Clara Smal, Diego S Vazquez, María Georgina Herrera, Ernesto A Roman, Nadine Alaimo, Mariana Gallo, Javier Santos
Human frataxin (FXN) is a highly conserved mitochondrial protein involved in iron homeostasis and activation of the iron-sulfur cluster assembly. FXN deficiency causes the neurodegenerative disease Friedreich's Ataxia. Here, we investigated the effect of alterations in loop-1, a stretch presumably essential for FXN function, on the conformational stability and dynamics of the native state. We generated four loop-1 variants, carrying substitutions, insertions and deletions. All of them were stable and well-folded proteins...
October 30, 2017: Archives of Biochemistry and Biophysics
https://www.readbyqxmd.com/read/29094619/acute-in-vitro-and-in-vivo-toxicity-of-a-commercial-grade-boron-nitride-nanotube-mixture
#8
Vamsi K Kodali, Jenny R Roberts, Mohammad Shoeb, Michael G Wolfarth, Lindsey Bishop, Tracy Eye, Mark Barger, Katherine A Roach, Sherri Friend, Diane Schwegler-Berry, Bean T Chen, Aleksandr Stefaniak, Kevin C Jordan, Roy R Whitney, Dale W Porter, Aaron D Erdely
Boron nitride nanotubes (BNNTs) are an emerging engineered nanomaterial attracting significant attention due to superior electrical, chemical and thermal properties. Currently, the toxicity profile of this material is largely unknown. Commercial grade BNNTs are composed of a mixture (BNNT-M) of ∼50-60% BNNTs, and ∼40-50% impurities of boron and hexagonal boron nitride. We performed acute in vitro and in vivo studies with commercial grade BNNT-M, dispersed by sonication in vehicle, in comparison to the extensively studied multiwalled carbon nanotube-7 (MWCNT-7)...
October 2017: Nanotoxicology
https://www.readbyqxmd.com/read/29093165/sequences-flanking-the-transmembrane-segments-facilitate-mitochondrial-localization-and-membrane-fusion-by-mitofusin
#9
Xiaofang Huang, Xin Zhou, Xiaoyu Hu, Amit S Joshi, Xiangyang Guo, Yushan Zhu, Quan Chen, William A Prinz, Junjie Hu
Mitochondria constantly divide and fuse. Homotypic fusion of the outer mitochondrial membranes requires the mitofusin (MFN) proteins, a family of dynamin-like GTPases. MFNs are anchored in the membrane by transmembrane (TM) segments, exposing both the N-terminal GTPase domain and the C-terminal tail (CT) to the cytosol. This arrangement is very similar to that of the atlastin (ATL) GTPases, which mediate fusion of endoplasmic reticulum (ER) membranes. We engineered various MFN-ATL chimeras to gain mechanistic insight into MFN-mediated fusion...
November 1, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29080362/metabolic-phenotyping-of-cho-cells-varying-in-cellular-biomass-accumulation-and-maintenance-during-fed-batch-culture
#10
Alejandro Fernandez-Martell, Yusuf B Johari, David C James
CHO cell lines capable of high-level recombinant protein product biosynthesis during fed-batch culture are still generally obtained by intensive empirical screening of transfected cells rather than knowledge-guided cellular engineering. In this study, we investigate how CHO cell lines create and maintain cellular biosynthetic capacity during fed-batch culture to achieve the optimal combination of rapid exponential proliferation and extended maintenance of high cell biomass concentration. We perform a comparative meta-analysis of mitochondrial and glycolytic functions of 22 discrete parental CHO cell lineages varying in fed-batch culture performance to test the hypotheses that (i) "biomass-intensive" CHO cells exhibit conserved differences in metabolic programming and (ii) it is possible to isolate parental CHO cell lines with a biomass-intensive phenotype to support fed-batch bioproduction processes...
October 28, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29079356/parkin-overexpression-in-human-mesenchymal-stromal-cells-from-wharton-s-jelly-suppresses-6-hydroxydopamine-induced-apoptosis-potential-therapeutic-strategy-in-parkinson-s-disease
#11
A R Bonilla-Porras, A Arevalo-Arbelaez, J F Alzate-Restrepo, C Velez-Pardo, M Jimenez-Del-Rio
BACKGROUND AIMS: Stem cell transplantation is an excellent option for regenerative or replacement therapy. However, deleterious microenvironmental and endogenous factors (e.g., oxidative stress) compromise ongoing graft survival and longevity. Therefore, (transient or stable) genetically modified cells may be reasonably thought to resist oxidative stress-induced damage. Genetic engineering of mesenchymal stromal cells (MSCs) obtained from Wharton's jelly tissue may offer some therapeutic potential...
October 24, 2017: Cytotherapy
https://www.readbyqxmd.com/read/29072519/determination-of-genetic-variations-in-the-genus-dryomys-thomas-1906-rodentia-gliridae-distributed-in-turkey-using-nadh-dehydrogenase-1-nd1-gene
#12
Teoman Kankılıç, Perinçek Seçkinozan Şeker, Arif Can Erdik, Tolga Kankılıç, Engin Selvi, Nuri Yiğit, Ercüment Çolak
Genetic diversity and phylogeny of Dryomys nitedula and Dryomys laniger from Turkey was described in the present study by using mitochondrial DNA NADH dehydrogenase 1 gene (ND1). Genetic variation in ND1 gene was determined by two model-based phylogenetic analyses and a network analysis revealed 27 haplotypes of D. nitedula constructing four main lineages (Thrace, Anatolia, North-eastern Anatolia and Şavşat) that have non-overlapping geographic distributions and no shared haplotypes, but on the other hand, three haplotypes were detected in four samples of D...
October 26, 2017: Mitochondrial DNA. Part A. DNA Mapping, Sequencing, and Analysis
https://www.readbyqxmd.com/read/29070706/does-egg-donation-for-mitochondrial-replacement-techniques-generate-parental-responsibilities
#13
César Palacios-González
Children created through mitochondrial replacement techniques (MRTs) are commonly presented as possessing 50% of their mother's nuclear DNA, 50% of their father's nuclear DNA and the mitochondrial DNA of an egg donor. This lab-engineered genetic composition has prompted two questions: Do children who are the product of an MRT procedure have three genetic parents? And, do MRT egg donors have parental responsibilities for the children created? In this paper, I address the second question and in doing so I also address the first one...
October 25, 2017: Journal of Medical Ethics
https://www.readbyqxmd.com/read/29064010/toxicity-of-copper-oxide-cuo-nanoparticles-on-human-blood-lymphocytes
#14
Evelyn Assadian, Mohammad Hadi Zarei, Ali Ghanadzadeh Gilani, Mehrzad Farshin, Hamid Degampanah, Jalal Pourahmad
CuO nanoparticles (CuO-NPs) serve several important functions in human life, particularly in the fields of medicine, engineering, and technology. These nanoparticles have been utilized as catalysts, semiconductors, sensors, gaseous and solid ceramic pigments, and magnet rotatable devices. Further use for CuO-NPs has been employed in the pharmaceutical industry especially in the production of anti-microbial fabric treatments or prevention of infections caused by Escherichia coli and methicillin-resistant Staphylococcus aureus...
October 24, 2017: Biological Trace Element Research
https://www.readbyqxmd.com/read/29057653/mitochondrion-targeting-environment-sensitive-red-fluorescent-probe-for-highly-sensitive-detection-and-imaging-of-vicinal-dithiol-containing-proteins
#15
Feng Liu, Hai-Juan Liu, Xian-Jun Liu, Wen Chen, Fenglin Wang, Ru-Qin Yu, Jian-Hui Jiang
Mitochondrial vicinal dithiol-containing proteins (VDPs) are key regulators in cellular redox homeostasis and useful markers for diagnostics of redox-dependent diseases. Current probes fail to target mitochondrial VDPs and show limited sensitivity and response rate. We develop a novel fluorescent probe using an engineered benzoxadiazole fluorophore that allows selective targeting of mitochondria and exhibits highly sensitive environment responsiveness. This probe is almost nonfluorescent in aqueous media, while delivering intense fluorescence upon binding to VDPs via a cyclic dithiaarsane ligand...
November 7, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/29056456/re-creation-of-a-key-step-in-the-evolutionary-switch-from-c3-to-c4-leaf-anatomy
#16
Peng Wang, Roxana Khoshravesh, Shanta Karki, Ronald Tapia, C Paolo Balahadia, Anindya Bandyopadhyay, W Paul Quick, Robert Furbank, Tammy L Sage, Jane A Langdale
The C4 photosynthetic pathway accounts for ∼25% of primary productivity on the planet despite being used by only 3% of species. Because C4 plants are higher yielding than C3 plants, efforts are underway to introduce the C4 pathway into the C3 crop rice. This is an ambitious endeavor; however, the C4 pathway evolved from C3 on multiple independent occasions over the last 30 million years, and steps along the trajectory are evident in extant species. One approach toward engineering C4 rice is to recapitulate this trajectory, one of the first steps of which was a change in leaf anatomy...
November 6, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/29044478/enhanced-pyruvate-production-in-candida-glabrata-by-carrier-engineering
#17
Zhengshan Luo, Song Liu, Guocheng Du, Sha Xu, Jingwen Zhou, Jian Chen
Pyruvate is an important organic acid that plays a key role in the central metabolic pathway. Manipulating transporters is an efficient strategy to enhance production of target organic acids and a means to understand the effects of altered intracellular pyruvate content on global metabolic networks. Efforts have been made to manipulate mitochondrial pyruvate carrier (MPC) to transport pyruvate into different subcellular compartments in Candida glabrata to demonstrate the effects of the subcellular distribution of pyruvate on central carbon metabolism...
October 16, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29029479/the-genetically-engineered-drug-rhcnb-induces-apoptosis-via-a-mitochondrial-route-in-tumor-cells
#18
Yang Yang, Huan Yang, Jinju Yang, Li Li, Benqiong Xiang, Qun Wei
The calcineurin B subunit (CNB) has antitumor activity. We showed previously that recombinant human CNB (rhCNB) also had strong anti-tumor activity in vivo, and was thus a promising candidate anti-tumor drug. It appeared to kill tumor cells via immunomodulation. Here, we show that rhCNB inhibits the proliferation of human hepatoma HepG-2 cells, resulting in their apoptosis. Exogenous CNB was found to localize to mitochondria in tumor cells and activate the mitochondrial apoptosis pathway, as indicated by a decrease of mitochondrial transmembrane potential, release of cytochrome C and activation of caspase-9, which then activates caspase-3...
September 12, 2017: Oncotarget
https://www.readbyqxmd.com/read/28978108/tunicamycin-induced-endoplasmic-reticulum-stress-promotes-apoptosis-of-prostate-cancer-cells-by-activating-mtorc1
#19
Prasun Guha, Engin Kaptan, Padmaja Gade, Dhananjaya V Kalvakolanu, Hafiz Ahmed
Studies suggest that tunicamycin may work as a therapeutic drug to cancer cells by inducing stress in the endoplasmic reticulum (ER) through unfolded protein response (UPR) and thereby promoting apoptosis. However, mechanisms of the prolonged activation of the UPR under sustained ER stress in the regulation of cell apoptosis are largely unknown. To delineate the role of candidate genes in the apoptotic process under ER stress and to search for new therapeutic strategies to treat metastatic castration resistant prostate cancer, we performed whole genome expression microarray analysis in tunicamycin treated metastatic androgen-insensitive prostate cancer cells, PC-3...
September 15, 2017: Oncotarget
https://www.readbyqxmd.com/read/28961779/nanopore-sequencing-enables-near-complete-de-novo-assembly-of-saccharomyces-cerevisiae-reference-strain-cen-pk113-7d
#20
Alex N Salazar, Arthur R Gorter de Vries, Marcel van den Broek, Melanie Wijsman, Pilar de la Torre Cortés, Anja Brickwedde, Nick Brouwers, Jean-Marc G Daran, Thomas Abeel
The haploid Saccharomyces cerevisiae strain CEN.PK113-7D is a popular model system for metabolic engineering and systems biology research. Current genome assemblies are based on short-read sequencing data scaffolded based on homology to strain S288C. However, these assemblies contain large sequence gaps, particularly in subtelomeric regions, and the assumption of perfect homology to S288C for scaffolding introduces bias. In this study, we obtained a near-complete genome assembly of CEN.PK113-7D using only Oxford Nanopore Technology's MinION sequencing platform...
September 13, 2017: FEMS Yeast Research
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