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  • 1
    ISSN: 1432-2048
    Keywords: Bradyrhizobium ; Electron microscopy ; Glycine (root nodules) ; High-pressure freezing ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract High-pressure freezing of chemically untreated nodules of soybean (Glycine max (L.) Merr.), in sharp contrast to chemical fixation and prefixation, appears to preserve the ultrastructure close to the native state. This is supported by the observation that the peribacteroid membrane of high-pressure-frozen samples is tightly wrapped around the bacteroids, a finding that is fully consistent with the current views on the physiology of oxygen and metabolite transport between plant cytosol and bacteroids. In soybean root nodules, the plant tissue and the enclosed bacteria are so dissimilar that conventional aldehyde-fixation procedures are unable to preserve the overall native ultrastructure. This was demonstrated by high-pressure freezing of nodules that had been pre-fixed in glutaraldehyde at various buffer molalities: no buffer strength tested preserved all ultrastructural aspects that could be seen after high-pressure freezing of chemically untreated nodules.
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  • 2
    ISSN: 1432-072X
    Keywords: Bradyrhizobium ; Electron microscopy ; Mutants ; Nitrogen fixation ; Nodulation ; Soybean ; Symbiosis ; Transposon Tn5
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genome of the slow-growing Bradyrhizobium japonicum (strain 110) was mutagenized with transposon Tn5. A total of 1623 kanamycin/streptomycin resistant derivatives were screened in soybean infection tests for nodulation (Nod) and symbiotic nitrogen fixation (Fix). In this report we describe 14 strains possessing a stable, reproducible Nod+Fix- phenotype. These strains were also grown under microaerobic culture conditions to test them for free-living nitrogen fixation activity (Nif). In addition to strains having reduced Fix and Nif activities, there were also strains that had reduced symbiotic Fix activity but were Nif+ ex planta. Analysis of the genomic structure revealed that the majority of the strains had a single Tn5 insertion without any further apparent physical alteration. A few strains had additional insertions (by Tn5 or IS50), or a deletion, or had cointegrated part of the vector used for Tn5 mutagenesis. One of the insertions was found in a known nif gene (nifD) whereas all other mutations seem to affect different, hitherto unknown genes or operons. Several mutant strains had an altered nodulation phenotype, inducing numerous, small, widely distributed nodules. Light and electron microscopy revealed that most of these mutants were defective in different stages of bacteroid development and/or bacteroid persistence. The protein patterns of the mutants were inspected by two-dimensional gel electrophoresis after labelling microaerobic cultures with l-(35S)methionine. Of particular interest were mutants lacking a group of proteins the synthesis of which was known to be under oxygen control. Such strains can be regarded as potential regulatory mutants.
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  • 3
    ISSN: 1432-072X
    Keywords: Bradyrhizobium ; Gene cloning ; Heme ; Marker exchange mutagenesis ; Nitrogen fixation ; Respiration ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Random and site-directed Tn5-induced mutagenesis of Bradyrhizobium japonicum yielded two mutations, one in strain 2960 and the other in strain 2606::Tn5-20, which mapped close to each other but in separate genes. The corresponding wild-type genes were cloned, and their approximate location on the cloned DNA was determined. Mutant 2960 was Fix- and formed green nodules on soybean, whereas strain 2606::Tn5-20 had ca. 4% of wild-type Fix activity and formed white nodules. Cytochrome oxidase assays (Nadi tests) showed a negative reaction with both mutants, indicating a functional deficiency of cytochrome c or its terminal oxidase or both. However, the mutants grew well under aerobic conditions on minimal media with different carbon sources. Furthermore, mutant 2960 had a reduced activity in hydrogen uptake, was unable to grow anaerobically with nitrate as the terminal electron acceptor and 2960-infected soybean nodules contained little, if any, functional leghemoglobin. Southern blot analysis showed that a B. japonicum heme biosynthesis mutant [strain LO505: O'Brian MR, Kirshbom PM, Maier RJ (1987) Proc Natl Acad Sci USA 84: 8390–8393] had its mutation close to the Tn5 insertion site of our mutant 2606::Tn5-20. This finding, combined with the observed phenotypes, suggested that the genes affected in mutants 2960 and 2606::Tn5-20 were involved in some steps of heme biosynthesis thus explaining the pleiotropic respiratory deficiencies of the mutants. Similar to strain LO505, the mutant 2606::Tn5-20 (but not 2960) was defective in the activity of protoporphyrinogen IX oxidase which catalyzes the penultimate step in the heme biosynthesis pathway. This suggests that one of the two cloned genes may code for this enzyme.
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  • 4
    Publication Date: 2001-04-28
    Description: Embryonic stem (ES) cells are fully pluripotent in that they can differentiate into all cell types, including gametes. We have derived 35 ES cell lines via nuclear transfer (ntES cell lines) from adult mouse somatic cells of inbred, hybrid, and mutant strains. ntES cells contributed to an extensive variety of cell types, including dopaminergic and serotonergic neurons in vitro and germ cells in vivo. Cloning by transfer of ntES cell nuclei could result in normal development of fertile adults. These studies demonstrate the full pluripotency of ntES cells.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Wakayama, T -- Tabar, V -- Rodriguez, I -- Perry, A C -- Studer, L -- Mombaerts, P -- New York, N.Y. -- Science. 2001 Apr 27;292(5517):740-3.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉The Rockefeller University, New York, NY 10021, USA. teru@advancedcell.com〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/11326103" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Blastocyst/*cytology ; *Cell Differentiation ; Cell Line ; Cell Lineage ; Chimera ; Cloning, Organism ; Crosses, Genetic ; Dopamine/metabolism ; Embryo Transfer ; Female ; Germ Cells/*cytology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Mice, Inbred ICR ; Mice, Nude ; Neurons/*cytology ; *Nuclear Transfer Techniques ; Serotonin/metabolism ; Stem Cells/*cytology
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 5
    Publication Date: 2018-08-11
    Description: Author(s): Pascal Naubereit, Tina Gottwald, Dominik Studer, and Klaus Wendt We present high-resolution data of the single-excitation spectrum of protactinium, reaching slightly beyond the first-ionization threshold. Within this work, more than 1500 energy levels are recorded in different excitation energy ranges below 50 000 cm − 1 . Our experimental results show that the tabula... [Phys. Rev. A 98, 022505] Published Fri Aug 10, 2018
    Keywords: Atomic and molecular structure and dynamics ; high-precision measurements
    Print ISSN: 1050-2947
    Electronic ISSN: 1094-1622
    Topics: Physics
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  • 6
    Publication Date: 2009-08-21
    Description: The isolation of human induced pluripotent stem cells (iPSCs) offers a new strategy for modelling human disease. Recent studies have reported the derivation and differentiation of disease-specific human iPSCs. However, a key challenge in the field is the demonstration of disease-related phenotypes and the ability to model pathogenesis and treatment of disease in iPSCs. Familial dysautonomia (FD) is a rare but fatal peripheral neuropathy, caused by a point mutation in the IKBKAP gene involved in transcriptional elongation. The disease is characterized by the depletion of autonomic and sensory neurons. The specificity to the peripheral nervous system and the mechanism of neuron loss in FD are poorly understood owing to the lack of an appropriate model system. Here we report the derivation of patient-specific FD-iPSCs and the directed differentiation into cells of all three germ layers including peripheral neurons. Gene expression analysis in purified FD-iPSC-derived lineages demonstrates tissue-specific mis-splicing of IKBKAP in vitro. Patient-specific neural crest precursors express particularly low levels of normal IKBKAP transcript, suggesting a mechanism for disease specificity. FD pathogenesis is further characterized by transcriptome analysis and cell-based assays revealing marked defects in neurogenic differentiation and migration behaviour. Furthermore, we use FD-iPSCs for validating the potency of candidate drugs in reversing aberrant splicing and ameliorating neuronal differentiation and migration. Our study illustrates the promise of iPSC technology for gaining new insights into human disease pathogenesis and treatment.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2784695/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2784695/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Lee, Gabsang -- Papapetrou, Eirini P -- Kim, Hyesoo -- Chambers, Stuart M -- Tomishima, Mark J -- Fasano, Christopher A -- Ganat, Yosif M -- Menon, Jayanthi -- Shimizu, Fumiko -- Viale, Agnes -- Tabar, Viviane -- Sadelain, Michel -- Studer, Lorenz -- R01 NS052671/NS/NINDS NIH HHS/ -- R01 NS052671-03/NS/NINDS NIH HHS/ -- England -- Nature. 2009 Sep 17;461(7262):402-6. doi: 10.1038/nature08320. Epub 2009 Aug 19.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Developmental Biology Program, Sloan-Kettering Institute, 1275 York Ave, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/19693009" target="_blank"〉PubMed〈/a〉
    Keywords: Adolescent ; Alternative Splicing/drug effects/genetics ; Animals ; Carrier Proteins/genetics ; Cell Dedifferentiation ; Cell Differentiation ; Cell Lineage ; Cell Movement ; Cells, Cultured ; Child ; Dysautonomia, Familial/drug therapy/genetics/*pathology/*therapy ; Female ; Fibroblasts/cytology/metabolism ; Gene Expression Profiling ; Humans ; Kinetin/pharmacology/therapeutic use ; Male ; Mice ; *Models, Biological ; Neural Crest/cytology/drug effects ; Organ Specificity ; Phenotype ; Pluripotent Stem Cells/cytology/drug effects/*metabolism/*transplantation
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 7
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    Nature Publishing Group (NPG)
    Publication Date: 2010-11-19
    Description: 〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Studer, Lorenz -- England -- Nature. 2010 Nov 18;468(7322):383-4. doi: 10.1038/468383a.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/21085168" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Brain/cytology/metabolism ; DNA Methylation ; Gene Silencing ; Humans ; Induced Pluripotent Stem Cells/metabolism ; Long Interspersed Nucleotide Elements/*genetics ; Methyl-CpG-Binding Protein 2/deficiency/genetics/*metabolism ; Mice ; Neuroepithelial Cells/metabolism ; Neurons/*metabolism ; Recombination, Genetic/*genetics ; Rett Syndrome/genetics/pathology ; Transcription, Genetic/genetics
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 8
    Publication Date: 2002-02-02
    Description: 〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Cibelli, Jose B -- Grant, Kathleen A -- Chapman, Karen B -- Cunniff, Kerrianne -- Worst, Travis -- Green, Heather L -- Walker, Stephen J -- Gutin, Philip H -- Vilner, Lucy -- Tabar, Viviane -- Dominko, Tanja -- Kane, Jeff -- Wettstein, Peter J -- Lanza, Robert P -- Studer, Lorenz -- Vrana, Kent E -- West, Michael D -- P50-AA11997/AA/NIAAA NIH HHS/ -- T32-AA07565/AA/NIAAA NIH HHS/ -- New York, N.Y. -- Science. 2002 Feb 1;295(5556):819.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Advanced Cell Technology, One Innovation Drive, Worcester, MA 01605, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/11823632" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Astrocytes/cytology ; Blastocyst/*cytology/physiology ; Cell Culture Techniques ; Cell Differentiation ; Cell Division ; Cell Line ; Cell Separation ; Cloning, Organism ; Dopamine/metabolism ; Embryo, Mammalian/*cytology ; Karyotyping ; *Macaca fascicularis ; Mice ; Mice, SCID ; Neurons/cytology ; *Parthenogenesis ; Serotonin/metabolism ; Stem Cells/*cytology/physiology ; Teratoma/pathology
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 9
    Publication Date: 2011-11-08
    Description: Human pluripotent stem cells (PSCs) are a promising source of cells for applications in regenerative medicine. Directed differentiation of PSCs into specialized cells such as spinal motoneurons or midbrain dopamine (DA) neurons has been achieved. However, the effective use of PSCs for cell therapy has lagged behind. Whereas mouse PSC-derived DA neurons have shown efficacy in models of Parkinson's disease, DA neurons from human PSCs generally show poor in vivo performance. There are also considerable safety concerns for PSCs related to their potential for teratoma formation or neural overgrowth. Here we present a novel floor-plate-based strategy for the derivation of human DA neurons that efficiently engraft in vivo, suggesting that past failures were due to incomplete specification rather than a specific vulnerability of the cells. Midbrain floor-plate precursors are derived from PSCs 11 days after exposure to small molecule activators of sonic hedgehog (SHH) and canonical WNT signalling. Engraftable midbrain DA neurons are obtained by day 25 and can be maintained in vitro for several months. Extensive molecular profiling, biochemical and electrophysiological data define developmental progression and confirm identity of PSC-derived midbrain DA neurons. In vivo survival and function is demonstrated in Parkinson's disease models using three host species. Long-term engraftment in 6-hydroxy-dopamine-lesioned mice and rats demonstrates robust survival of midbrain DA neurons derived from human embryonic stem (ES) cells, complete restoration of amphetamine-induced rotation behaviour and improvements in tests of forelimb use and akinesia. Finally, scalability is demonstrated by transplantation into parkinsonian monkeys. Excellent DA neuron survival, function and lack of neural overgrowth in the three animal models indicate promise for the development of cell-based therapies in Parkinson's disease.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3245796/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3245796/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Kriks, Sonja -- Shim, Jae-Won -- Piao, Jinghua -- Ganat, Yosif M -- Wakeman, Dustin R -- Xie, Zhong -- Carrillo-Reid, Luis -- Auyeung, Gordon -- Antonacci, Chris -- Buch, Amanda -- Yang, Lichuan -- Beal, M Flint -- Surmeier, D James -- Kordower, Jeffrey H -- Tabar, Viviane -- Studer, Lorenz -- NS052671/NS/NINDS NIH HHS/ -- P50 NS047085/NS/NINDS NIH HHS/ -- P50 NS071669/NS/NINDS NIH HHS/ -- P50 NS071669-03/NS/NINDS NIH HHS/ -- England -- Nature. 2011 Nov 6;480(7378):547-51. doi: 10.1038/nature10648.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Center for Stem Cell Biology, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, New York 10065, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/22056989" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; *Brain Tissue Transplantation ; Cell Differentiation ; Cell Line ; Cell Survival ; Dopaminergic Neurons/*cytology/*transplantation ; Embryonic Stem Cells/*cytology ; Female ; Humans ; Macaca mulatta ; Mesencephalon/cytology ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Parkinson Disease/*therapy ; Rats ; Rats, Sprague-Dawley
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 10
    Publication Date: 2018-08-11
    Description: Author(s): Pascal Naubereit, Dominik Studer, Anna V. Viatkina, Andreas Buchleitner, Barbara Dietz, Victor V. Flambaum, and Klaus Wendt Recently, spectroscopic investigations of the protactinium atom applying resonant laser ionization spectroscopy revealed high-resolution data of the single-excitation spectrum of protactinium, reaching slightly beyond the first ionization potential [P. Naubereit et al. , preceding paper, Phys. Rev. ... [Phys. Rev. A 98, 022506] Published Fri Aug 10, 2018
    Keywords: Atomic and molecular structure and dynamics ; high-precision measurements
    Print ISSN: 1050-2947
    Electronic ISSN: 1094-1622
    Topics: Physics
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