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Hong Lin, Jordi Magrane, Amy Rattelle, Anna Stepanova, Alexander Galkin, Elisia M Clark, Yi Na Dong, Sarah M Halawani, David R Lynch
Friedreich ataxia (FRDA), the most common recessive inherited ataxia, results from deficiency of frataxin, a small mitochondrial protein crucial for iron-sulphur cluster formation and ATP production. Frataxin deficiency is associated with mitochondrial dysfunction in FRDA patients and animal models; however, early mitochondrial pathology in FRDA cerebellum remains elusive. Using frataxin knock-in/knockout (KIKO) mice and KIKO mice carrying the mitoDendra transgene, we show early cerebellar deficits in mitochondrial biogenesis and respiratory chain complexes in this FRDA model...
November 1, 2017: Disease Models & Mechanisms
Patrick K McCamphill, Larissa Ferguson, Wayne S Sossin
Mechanistic target of rapamycin complex 1 (mTORC1)-dependent protein synthesis is required for many forms of synaptic plasticity and memory, but the downstream pathways important for synaptic plasticity are poorly understood. Long-term facilitation (LTF) in Aplysia is a form of synaptic plasticity that is closely linked to behavioral memory and an attractive model system for examining the important downstream targets for mTORC1 in regulating synaptic plasticity. Although mTORC1-regulated protein synthesis has been strongly linked to translation initiation, translation elongation is also regulated by mTORC1 and LTF leads to an mTORC1-dependent decrease in eukaryotic elongation factor 2 (eEF2) phosphorylation...
July 2017: Journal of Neurochemistry
Yair Lotan, Jodi Antonelli, Inmaculada Buendia Jiménez, Hakam Gharbi, Ron Herring, Allison Beaver, Aphrihl Dennis, Dendra Von Merveldt, Suzie Carter, Adam Cohen, John Poindexter, Orson W Moe, Margaret S Pearle
Preventing dehydration in subjects at risk may provide a means of primary prevention of kidney stones. The purpose of this pilot study was to assess the hydration status of an at-risk group of steel plant workers based on end-of-shift ('post-shift') spot urine osmolality and 24-h urinary stone risk parameters. 100 volunteers were recruited from Gerdau Midlothian steel mill in Texas on 11/14/14 and 12/5/14. Clinical data were recorded and post-shift spot urine sample was used to measure urine osmolality. Participants were invited to submit a 24-h urine sample within 4 weeks of enrollment...
April 2017: Urolithiasis
Jae-Hun Jeoung, Sebastian Goetzl, Sandra Elisabeth Hennig, Jochen Fesseler, Christina Wörmann, Julia Dendra, Holger Dobbek
The reductive acetyl-coenzyme A (acetyl-CoA) pathway, also known as the Wood-Ljungdahl pathway, allows reduction and condensation of two molecules of carbon dioxide (CO2) to build the acetyl-group of acetyl-CoA. Productive utilization of CO2 relies on a set of oxygen sensitive metalloenzymes exploiting the metal organic chemistry of nickel and cobalt to synthesize acetyl-CoA from activated one-carbon compounds. In addition to the central catalysts, CO dehydrogenase and acetyl-CoA synthase, ATPases are needed in the pathway...
May 2014: Biological Chemistry
Jordi Magrané, Czrina Cortez, Wen-Biao Gan, Giovanni Manfredi
Neuronal mitochondrial morphology abnormalities occur in models of familial amyotrophic lateral sclerosis (ALS) associated with SOD1 and TDP43 mutations. These abnormalities have been linked to mitochondrial axonal transport defects, but the temporal and spatial relationship between mitochondrial morphology and transport alterations in these two distinct genetic forms of ALS has not been investigated in vivo. To address this question, we crossed SOD1 (wild-type SOD1(WT) and mutant SOD1(G93A)) or TDP43 (mutant TDP43(A315T)) transgenic mice with mice expressing the fluorescent protein Dendra targeted to mitochondria in neurons (mitoDendra)...
March 15, 2014: Human Molecular Genetics
Youngnam N Jin, Yanxun V Yu, Soner Gundemir, Chulman Jo, Mei Cui, Kim Tieu, Gail V W Johnson
Huntington disease (HD) is an inherited neurodegenerative disease resulting from an abnormal expansion of polyglutamine in huntingtin (Htt). Compromised oxidative stress defense systems have emerged as a contributing factor to the pathogenesis of HD. Indeed activation of the Nrf2 pathway, which plays a prominent role in mediating antioxidant responses, has been considered as a therapeutic strategy for the treatment of HD. Given the fact that there is an interrelationship between impairments in mitochondrial dynamics and increased oxidative stress, in this present study we examined the effect of mutant Htt (mHtt) on these two parameters...
2013: PloS One
Verónica A Lombardo, Anje Sporbert, Salim Abdelilah-Seyfried
Embryogenesis is a dynamic process that is best studied by using techniques that allow the documentation of developmental changes in vivo. The use of genetically-encoded fluorescent proteins has proven a valuable strategy for elucidating dynamic morphogenetic processes as they occur in the intact organism. During the past decade, the development of photoactivatable and photoconvertible fluorescent proteins has opened the possibility to investigate the fate of discrete subpopulations of tagged proteins. Unlike photoactivatable proteins, photoconvertible fluorescent proteins (PCFPs) are readily tracked and imaged in their native emission state prior to photoconversion, making it easier to identify and select regions by optical inspection...
2012: Journal of Visualized Experiments: JoVE
Viktor Karlovitch Lund, Yvonne DeLotto, Robert DeLotto
We have assembled a molecular toolkit enabling the facile generation of C-terminal fluorescent protein fusion constructs for the construction of transgenic Drosophila lines in the form of a series of modified vectors for P-element transformation. These vectors contain one of a variety of fluorescent tags including GFP, mCherry, Venus, Cerulean, photoactivatable paGFP and the red-to-green photoconvertible protein Dendra and provide a range of options with respect to transcriptional regulation. The vectors have been extensively tested in vivo and can produce fluorescent chimeric proteins that are functional...
July 2011: Fly
Youmna Kfoury, Niclas Setterblad, Marwan El-Sabban, Alessia Zamborlini, Zeina Dassouki, Hiba El Hajj, Olivier Hermine, Claudine Pique, Hugues de Thé, Ali Saïb, Ali Bazarbachi
The human T-lymphotropic virus type I oncoprotein Tax is critical for T-cell transformation, acting mainly through nuclear factor kappa B essential modulator (NEMO) binding and subsequent nuclear factor-κB activation. Tax localizes to Tax nuclear bodies and to the centrosome and is subjected to ubiquitylation and small ubiquitin-like modifier (SUMO)ylation, which are both necessary for complete transcriptional activation. Using the photoconvertible fluorophore Dendra-2 coupled with live video confocal microscopy, we show for the first time that the same Tax molecule shuttles among Tax nuclear bodies and between these nuclear bodies and the centrosome, depending on its posttranslational modifications...
January 6, 2011: Blood
Susan M Baker, Robert W Buckheit, Matthias M Falk
BACKGROUND: Green fluorescent protein (GFP) and other FP fusions have been extensively utilized to track protein dynamics in living cells. Recently, development of photoactivatable, photoswitchable and photoconvertible fluorescent proteins (PAFPs) has made it possible to investigate the fate of discrete subpopulations of tagged proteins. Initial limitations to their use (due to their tetrameric nature) were overcome when monomeric variants, such as Dendra, mEos, and mKikGR were cloned/engineered...
February 22, 2010: BMC Cell Biology
Aristides B Arrenberg, Filippo Del Bene, Herwig Baier
Expression of halorhodopsin (NpHR), a light-driven microbial chloride pump, enables optical control of membrane potential and reversible silencing of targeted neurons. We generated transgenic zebrafish expressing enhanced NpHR under control of the Gal4/UAS system. Electrophysiological recordings showed that eNpHR stimulation effectively suppressed spiking of single neurons in vivo. Applying light through thin optic fibers positioned above the head of a semi-restrained zebrafish larva enabled us to target groups of neurons and to simultaneously test the effect of their silencing on behavior...
October 20, 2009: Proceedings of the National Academy of Sciences of the United States of America
Kevin C Flynn, Chi W Pak, Alisa E Shaw, Frank Bradke, James R Bamburg
Axonogenesis involves a shift from uniform delivery of materials to all neurites to preferential delivery to the putative axon, supporting its more rapid extension. Waves, growth cone-like structures that propagate down the length of neurites, were shown previously to correlate with neurite growth in dissociated cultured hippocampal neurons. Waves are similar to growth cones in their structure, composition and dynamics. Here, we report that waves form in all undifferentiated neurites, but occur more frequently in the future axon during initial neuronal polarization...
October 2009: Developmental Neurobiology
Joseph Selvin
To explore the functional role of the antagonistic producer strain Streptomyces dendra sp. nov. MSI051 in the host sponge Dendrilla nigra, hypothetical factors including the antagonistic potential of MSI051 against biofilm bacteria and a ubiquitous defense enzyme phospholipase A2 (PLA2) in host sponge as well as in bacterial symbiont MSI051 were determined. The host sponge D. nigra and associated bacterial symbiont MSI051 contained high levels of PLA2. The host sponge showed PLA2 activity to the extent of 1032 U/L, with a specific activity of 2021 U/g, and strain MSI051 showed similar activity...
May 2009: Current Microbiology
Nadya G Gurskaya, Vladislav V Verkhusha, Alexander S Shcheglov, Dmitry B Staroverov, Tatyana V Chepurnykh, Arkady F Fradkov, Sergey Lukyanov, Konstantin A Lukyanov
Green fluorescent protein (GFP) and GFP-like proteins represent invaluable genetically encoded fluorescent probes. In the last few years a new class of photoactivatable fluorescent proteins (PAFPs) capable of pronounced light-induced spectral changes have been developed. Except for tetrameric KFP1 (ref. 4), all known PAFPs, including PA-GFP, Kaede, EosFP, PS-CFP, Dronpa, PA-mRFP1 and KikGR require light in the UV-violet spectral region for activation through one-photon excitation--such light can be phototoxic to some biological systems...
April 2006: Nature Biotechnology
Javier Hernáandez, Samira Syed, Geoffrey Weiss, Gabriel Fernandes, Dendra von Merveldt, Dean A Troyer, Joseph W Basler, Ian M Thompson
PURPOSE: Studies suggest that vitamin E may decrease the risk of prostate cancer. The Prevention Research Veteran Affairs E-vitamin Nutrition Trial is a randomized, double-blind, placebo controlled study designed to assess the effects of vitamin E supplementation on biomarkers associated with prostate cancer risk in peripheral blood and prostate tissue. MATERIALS AND METHODS: A total of 44 patients with increased prostate specific antigen (PSA) and/or abnormal digital rectal examination on initial evaluation were randomized to receive 400 IU vitamin E (22) vs placebo (22)...
August 2005: Journal of Urology
Ian M Thompson, Joseph Basler, Deborah Hensley, Dendra von Merveldt, Carol A Jenkins, Betsy Higgins, Robin Leach, Dean Troyer, Brad Pollock
Prostate cancer prevention is now one of the most aggressively investigated areas of urologic oncology, with > 30,000 men currently participating in clinical trials in the United States alone. The Prostate Cancer Prevention Trial will complete end-of-study prostate biopsies in May 2004, and the Selenium and Vitamin E Cancer Prevention Trial is rapidly reaching its accrual goal 1-2 years ahead of schedule. These 2 studies will give definitive answers regarding 3 of the most important potential preventive interventions: finasteride, vitamin E, and selenium...
March 2003: Clinical Prostate Cancer
J Kress, A Rosner, A Hirsch
The convergent synthesis of a new class of chiral dendrimers is described. Owing to their structural resemblance to depsipeptides they are called depsipeptide dendrimers. The ex-chiral pool synthesis starts from (R,R)-, (S,S)-, and meso-tartaric acid as branching units and dipeptides or tripeptides consisting of glycine, (L)-alanine, and (L)-leucine as chiral-spacer building blocks. The key intermediates for the convergent assembly of such depsipeptide dendrimers are the peptide-tartaric acid conjugates 13a,b, 19a,b, 25, and 27, which contain either an unprotected C terminus of the peptide chain (13 a,b, 25) or two unprotected hydroxy groups within the tartaric acid termini...
January 2000: Chemistry: a European Journal
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