ALS spinal neurons show varied and reduced mtDNA gene copy numbers and increased mtDNA gene deletions

Background Spinal cord neurons of ALS patients demonstrate reduced cytochrome oxidase histochemical activity, and ALS spinal cord tissues have increased mitochondrial DNA (mtDNA) point mutations and depleted mtDNA levels. It is presently unknown whether mtDNA abnormalities are present in single human ALS neurons. Results Using laser capture microdissection (LCM) we isolated several hundred individual anterior spinal neurons from unfixed, frozen sections of 10 ALS and 7 age-matched CTL cervical spinal cords. DNA from each individual neuron was analyzed with multiplex qPCR for ND2, CO3, and ND4, three mitochondrial DNA genes encoding respiratory proteins. Scatterplots of individual spinal neuron results showed extensive heterogeneity of mtDNA gene levels across 4-5 orders of magnitude that were much more clustered in single Purkinje neurons isolated from CTL cerebella. Plots of ratios of ND4/ND2 and CO3/ND2 showed that many but not all ALS neurons from individuals contained low ratios of these mtDNA genes, implying greater abundances of mtDNA deletions in the major arc. Single CTL cerebellar Purkinje neurons did not contain high levels of apparent mtDNA deletions observed in anterior spinal neurons. Conclusions At the time of ALS subjects\u27 deaths, many but not all surviving anterior neurons in their cervical spinal cords have reduced mtDNA gene levels and increased mtDNA deletion abundances that arise for unclear reasons. If these anterior spinal neuron mtDNA gene deficiencies contribute to bioenergetic impairments, reduced synaptic function and increased risk of degeneration, then introduction into mitochondria and expression of intact mtDNA, now available through use of recently developed recombinant human TFAM, may reverse the course of ALS

Coherent control and feedback cooling in a remotely-coupled hybrid atom-optomechanical system

Cooling to the motional ground state is an important first step in the preparation of nonclassical states of mesoscopic mechanical oscillators. Light-mediated coupling to a remote atomic ensemble has been proposed as a method to reach the ground state for low frequency oscillators. The ground state can also be reached using optical measurement followed by feedback control. Here we investigate the possibility of enhanced cooling by combining these two approaches. The combination, in general, outperforms either individual technique, though atomic ensemble-based cooling and feedback cooling each individually dominate over large regions of parameter space.Comment: 28 pages, 5 figures, 2 tables. Updated to include exemplary experimental parameters and expanded discussion of noise source

A preliminary list of the rusts of Boone County

Approved May 15, 1913, George M. ReedTypescriptPhotographs pasted inM.A. University of Missouri 1913A complete list of the rusts of Boone County has not been published. In the year 1889 Dr. B.T. Galloway of the U. S. Department of Agriculture, published a preliminary list of the parasitic fungi of Missouri. The specimens which Dr. Galloway collected were destroyed at the time of the fire which demolished the main building of the University in February, 1892. This list included one hundred and two rusts, only three of which are mentioned specifically as having been collected in Boone County. Twenty-six of the list are mentioned as common forms, and were probably found in this immediate vicinity. The larger number of the remainder of the forms were collected in Perry County, located on the Mississippi river in the southeastern part of the state. Since that date a considerable number of specimens has been collected, largely from this region, and placed in the herbarium of the University. The collection contains a total of four hundred and forty-three specimens. Most of these were collected on general field trips taken by different members of the Department of Botany, in the years 1910, 1911, and 1912, but no systematic collection of Rusts has been undertaken. It has been the purpose of the present work to identify the rusts occurring in the collection and arrange them in order according to the classification accepted at the present time. This collection and arrangement may then serve as a basis for further work. The present list doubtless includes but a small portion of the rusts to be found in this region, and the addition to the collection of other forms from this locality, as well as from the remainder of the state, is very desirable. It is quite probably that the rust flora of Missouri is as abundant and diversified as that of any other state.Includes bibliographical reference

A two-year field study with transgenic Bacillus thuringiensis maize: effects on soil microorganisms

We evaluated the changes of some soil microbiological characteristics due to the use of transgenic maize expressing Bacillus thuringiensis (Bt) toxin. A two-year field experiment was conducted (2003 and 2004). Two lines of transgenic Bt maize that express the Cry1Ab protein (event 176 and MON 810) and their near-isogenic non-Bt lines were used. Rhizosphere and non-rhizosphere soils were collected and measurements were performed during the maize cultural cycle and immediately at pre-harvest. Key soil microbiological parameters measured included the numbers of culturable aerobic bacteria, including actinomycetes, and fungi, the activity of dehydrogenase and nitrogenase enzymes and ATP content. There were clear seasonal effects in the microbial parameters as evidenced by the consistent changes in sampling dates across the two years. Differences in the measured variables were also observed between rhizosphere and non-rhizosphere soils. However, under our field conditions, the presence of Bt maize did not cause, in a general way, changes in the microbial populations of the soil or in the activity of the microbial communit

Recombinant Mitochondrial Transcription Factor A with N-terminal Mitochondrial Transduction Domain Increases Respiration and Mitochondrial Gene Expression in G11778A Leber's Hereditary Optic Neuropathy Cybrid Cells

Diseases involving mitochondrial defects usually manifest themselves in high-energy, post-mitotic tissues such as brain, retina, skeletal and cardiac muscle and frequently cause deficiencies in mitochondrial bioenergetics. We have developed a scalable procedure to produce recombinant human mitochondrial transcription factor A (TFAM) modified with an N-terminal protein transduction domain (PTD) and mitochondrial localization signal (MLS) that allow it to cross membranes and enter mitochondria through its "mitochondrial transduction domain" (MTD,=PTD+MLS). _In vitro_ studies in a classic mitochondrial disease cell model demonstrated that Alexa488-labeled MTD-TFAM rapidly entered the mitochondrial compartment. MTD-TFAM treatment of these cell lines reversibly increased oxygen consumption (respiration) rates 3-fold, levels of respiratory proteins and mitochondrial gene expression. _In vivo_ results demonstrated that respiration increased to lesser degrees in mitochondria from tissues of mice injected with MTD-TFAM. MTD-TFAM can alter mitochondrial bioenergetics and holds promise for treatment of mitochondrial diseases involving deficiencies of energy production

Continuous optical-to-mechanical quantum state transfer in the unresolved sideband regime

Optical-to-mechanical quantum state transfer is an important capability for future quantum networks, quantum communication, and distributed quantum sensing. However, existing continuous state transfer protocols operate in the resolved sideband regime, necessitating a high-quality optical cavity and a high mechanical resonance frequency. Here, we propose a continuous protocol that operates in the unresolved sideband regime. The protocol is based on feedback cooling, can be implemented with current technology, and is able to transfer non-Gaussian quantum states with high fidelity. Our protocol significantly expands the kinds of optomechanical devices for which continuous optical-to-mechanical state transfer is possible, paving the way towards quantum technological applications and the preparation of macroscopic superpositions to test the fundamentals of quantum science

Epigenetic Modifications of the PGC-1α Promoter during Exercise Induced Expression in Mice

The transcriptional coactivator, PGC-1α, is known for its role in mitochondrial biogenesis. Although originally thought to exist as a single protein isoform, recent studies have identified additional promoters which produce multiple mRNA transcripts. One of these promoters (promoter B), approximately 13.7kb upstream of the canonical PGC-1α promoter (promoter A), yields alternative transcripts present at levels much lower than the canonical PGC-1α mRNA transcript. In skeletal muscle, exercise resulted in a substantial, rapid increase of mRNA of these alternative PGC-1α transcripts. Although the β2-adrenergic receptor was identified as a signaling pathway that activates transcription from PGC-1α promoter B, it is not yet known what molecular changes occur to facilitate PGC-1α promoter B activation following exercise. We sought to determine whether epigenetic modifications were involved in this exercise response in mouse skeletal muscle. We found that DNA hydroxymethylation correlated to increased basal mRNA levels from PGC-1α promoter A, but that DNA methylation appeared to play no role in the exercise-induced activation of PGC-1α promoter B. The level of the activating histone mark H3K4me3 increased with exercise 2–4 fold across PGC- 1α promoter B, but remained unaltered past the canonical PGC-1α transcriptional start site. Together, these data show that epigenetic modifications partially explain exercise-induced changes in the skeletal muscle mRNA levels of PGC-1α isoforms