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Sample records for maintenance protein tam41

  1. Fanconi anemia proteins in telomere maintenance.

    PubMed

    Sarkar, Jaya; Liu, Yie

    2016-07-01

    Mammalian chromosome ends are protected by nucleoprotein structures called telomeres. Telomeres ensure genome stability by preventing chromosome termini from being recognized as DNA damage. Telomere length homeostasis is inevitable for telomere maintenance because critical shortening or over-lengthening of telomeres may lead to DNA damage response or delay in DNA replication, and hence genome instability. Due to their repetitive DNA sequence, unique architecture, bound shelterin proteins, and high propensity to form alternate/secondary DNA structures, telomeres are like common fragile sites and pose an inherent challenge to the progression of DNA replication, repair, and recombination apparatus. It is conceivable that longer the telomeres are, greater is the severity of such challenges. Recent studies have linked excessively long telomeres with increased tumorigenesis. Here we discuss telomere abnormalities in a rare recessive chromosomal instability disorder called Fanconi Anemia and the role of the Fanconi Anemia pathway in telomere biology. Reports suggest that Fanconi Anemia proteins play a role in maintaining long telomeres, including processing telomeric joint molecule intermediates. We speculate that ablation of the Fanconi Anemia pathway would lead to inadequate aberrant structural barrier resolution at excessively long telomeres, thereby causing replicative burden on the cell. Published by Elsevier B.V.

  2. Getting Folded: Chaperone proteins in muscle development, maintenance and disease

    PubMed Central

    Smith, Daniel A.; Carland, Carmen R.; Guo, Yiming; Bernstein, Sanford I.

    2014-01-01

    Chaperone proteins are critical for protein folding and stability, and hence are necessary for normal cellular organization and function. Recent studies have begun to interrogate the role of this specialized class of proteins in muscle biology. During development, chaperone-mediated folding of client proteins enables their integration into nascent sarcomeres. In addition to assisting with muscle differentiation, chaperones play a key role in maintenance of muscle tissues. Further, disruption of the chaperone network can result in neuromuscular disease. In this review, we discuss how chaperones are involved in myofibrillogenesis, sarcomere maintenance and muscle disorders. We also consider the possibilities of therapeutically targeting chaperones to treat muscle disease. PMID:25125177

  3. Unwinding protein complexes in ALTernative telomere maintenance.

    PubMed

    Bhattacharyya, Saumitri; Sandy, April; Groden, Joanna

    2010-01-01

    Telomeres are composed of specialized chromatin that includes DNA repair/recombination proteins, telomere DNA-binding proteins and a number of three dimensional nucleic acid structures including G-quartets and D-loops. A number of studies suggest that the BLM and WRN recQ-like helicases play important roles in recombination-mediated mechanisms of telomere elongation or Alternative Lengthening of Telomeres (ALT), processes that maintain/elongate telomeres in the absence of telomerase. BLM and WRN localize within ALT-associated nuclear bodies in telomerase-negative immortalized cell lines and interact with the telomere-specific proteins POT1, TRF1 and TRF2. Helicase activity is modulated by these interactions. BLM functions in DNA double-strand break repair processes such as non-homologous end joining, homologous recombination-mediated repair, resolution of stalled replication forks and synthesis-dependent strand annealing, although its precise functions at the telomeres are speculative. WRN also functions in DNA replication, recombination and repair, and in addition to its helicase domain, includes an exonuclease domain not found in other recQ-like helicases. The biochemical properties of BLM and WRN are, therefore, important in biological processes other than DNA replication, recombination and repair. In this review, we discuss some previous and recent findings of human rec-Q-like helicases and their role in telomere elongation during ALT processes.

  4. Protein diets, body weight loss and weight maintenance.

    PubMed

    Martens, Eveline A P; Westerterp-Plantenga, Margriet S

    2014-01-01

    The review addresses briefly the relevance of protein diets for body weight loss and weight maintenance. The addition of recent findings on age-dependent protein requirements, specific effects of protein intake and protein source, the relevance of the other dietary macronutrients, especially of 'low-carb', 'protein leverage', the mechanisms of protein-induced satiety, and food-reward makes the review up-to-date. Different effects of protein diets in different age groups result from age-dependent protein requirements that are primarily related to effects on body composition. A protein intake of 0.8 g/kg/day is sufficient to sustain a negative energy balance in adults, irrespective of the protein source. 'Low-carb' diets trace back to the protein-induced effects. Evidence that protein intake drives energy intake as suggested by the 'Protein leverage hypothesis' is scarce and equivocal. Finally, limited protein-induced food reward may affect compliance to a protein diet. An implication of the findings for clinical practice is that a protein intake of 0.8-1.2 g/kg/day is sufficient to sustain satiety, energy expenditure, and fat-free mass, independent of a dietary 'low-carb' content. Limited protein-induced food reward may affect compliance to a protein diet.

  5. Protein interactions in genome maintenance as novel antibacterial targets.

    PubMed

    Marceau, Aimee H; Bernstein, Douglas A; Walsh, Brian W; Shapiro, Walker; Simmons, Lyle A; Keck, James L

    2013-01-01

    Antibacterial compounds typically act by directly inhibiting essential bacterial enzyme activities. Although this general mechanism of action has fueled traditional antibiotic discovery efforts for decades, new antibiotic development has not kept pace with the emergence of drug resistant bacterial strains. These limitations have severely restricted the therapeutic tools available for treating bacterial infections. Here we test an alternative antibacterial lead-compound identification strategy in which essential protein-protein interactions are targeted rather than enzymatic activities. Bacterial single-stranded DNA-binding proteins (SSBs) form conserved protein interaction "hubs" that are essential for recruiting many DNA replication, recombination, and repair proteins to SSB/DNA nucleoprotein substrates. Three small molecules that block SSB/protein interactions are shown to have antibacterial activity against diverse bacterial species. Consistent with a model in which the compounds target multiple SSB/protein interactions, treatment of Bacillus subtilis cultures with the compounds leads to rapid inhibition of DNA replication and recombination, and ultimately to cell death. The compounds also have unanticipated effects on protein synthesis that could be due to a previously unknown role for SSB/protein interactions in translation or to off-target effects. Our results highlight the potential of targeting protein-protein interactions, particularly those that mediate genome maintenance, as a powerful approach for identifying new antibacterial compounds.

  6. Biochemical function of mouse minichromosome maintenance 2 protein.

    PubMed

    Ishimi, Y; Komamura, Y; You, Z; Kimura, H

    1998-04-03

    Minichromosome maintenance (MCM) proteins play an essential role in eukaryotic DNA replication and bind to chromatin before the initiation of DNA replication. We reported that MCM protein complexes consisting of MCM2, -4, -6, and -7 bind strongly to a histone-Sepharose column (Ishimi, Y., Ichinose, S., Omori, A., Sato, K., and Kimura, H. (1996) J. Biol. Chem. 271, 24115-24122). Here, we have analyzed this interaction at the molecular level. We found that among six mouse MCM proteins, only MCM2 binds to histone; amino acid residues 63-153 are responsible for this binding. The region required for nuclear localization of MCM2 was mapped near this histone-binding domain. Far-Western blotting analysis of truncated forms of H3 histone indicated that amino acid residues 26-67 of H3 histone are required for binding to MCM2. We have also shown that mouse MCM2 can inhibit the DNA helicase activity of the human MCM4, -6, and -7 protein complex. These results suggest that MCM2 plays a different role in the initiation of DNA replication than the other MCM proteins.

  7. Axonal maintenance, glia, exosomes, and heat shock proteins

    PubMed Central

    Tytell, Michael; Lasek, Raymond J.; Gainer, Harold

    2016-01-01

    Of all cellular specializations, the axon is especially distinctive because it is a narrow cylinder of specialized cytoplasm called axoplasm with a length that may be orders of magnitude greater than the diameter of the cell body from which it originates. Thus, the volume of axoplasm can be much greater than the cytoplasm in the cell body. This fact raises a logistical problem with regard to axonal maintenance. Many of the components of axoplasm, such as soluble proteins and cytoskeleton, are slowly transported, taking weeks to months to travel the length of axons longer than a few millimeters after being synthesized in the cell body. Furthermore, this slow rate of supply suggests that the axon itself might not have the capacity to respond fast enough to compensate for damage to transported macromolecules. Such damage is likely in view of the mechanical fragility of an axon, especially those innervating the limbs, as rapid limb motion with high impact, like running, subjects the axons in the limbs to considerable mechanical force. Some researchers have suggested that local, intra-axonal protein synthesis is the answer to this problem. However, the translational state of axonal RNAs remains controversial. We suggest that glial cells, which envelop all axons, whether myelinated or not, are the local sources of replacement and repair macromolecules for long axons. The plausibility of this hypothesis is reinforced by reviewing several decades of work on glia-axon macromolecular transfer, together with recent investigations of exosomes and other extracellular vesicles, as vehicles for the transmission of membrane and cytoplasmic components from one cell to another. PMID:26962444

  8. Maintenance of native-like protein dynamics may not be required for engineering functional proteins.

    PubMed

    Gobeil, Sophie M C; Clouthier, Christopher M; Park, Jaeok; Gagné, Donald; Berghuis, Albert M; Doucet, Nicolas; Pelletier, Joelle N

    2014-10-23

    Proteins are dynamic systems, and understanding dynamics is critical for fully understanding protein function. Therefore, the question of whether laboratory engineering has an impact on protein dynamics is of general interest. Here, we demonstrate that two homologous, naturally evolved enzymes with high degrees of structural and functional conservation also exhibit conserved dynamics. Their similar set of slow timescale dynamics is highly restricted, consistent with evolutionary conservation of a functionally important feature. However, we also show that dynamics of a laboratory-engineered chimeric enzyme obtained by recombination of the two homologs exhibits striking difference on the millisecond timescale, despite function and high-resolution crystal structure (1.05 Å) being conserved. The laboratory-engineered chimera is thus functionally tolerant to modified dynamics on the timescale of catalytic turnover. Tolerance to dynamic variation implies that maintenance of native-like protein dynamics may not be required when engineering functional proteins.

  9. Blood profiling of proteins and steroids during weight maintenance with manipulation of dietary protein level and glycaemic index.

    PubMed

    Wang, Ping; Holst, Claus; Astrup, Arne; Bouwman, Freek G; van Otterdijk, Sanne; Wodzig, Will K W H; Andersen, Malene R; van Baak, Marleen A; Rasmussen, Lone G; Martinez, J Alfredo; Jebb, Susan A; Pfeiffer, Andreas F H; Kafatos, Anthony; Handjieva-Darlenska, Teodora; Hlavaty, Petr; Saris, Wim H M; Mariman, Edwin C M

    2012-01-01

    Weight regain after weight loss is common. In the Diogenes dietary intervention study, a high-protein and low-glycaemic index (GI) diet improved weight maintenance. The objective of the present study was to identify (1) blood profiles associated with continued weight loss and weight regain (2) blood biomarkers of dietary protein and GI levels during the weight-maintenance phase. Blood samples were collected at baseline, after 8 weeks of low-energy diet-induced weight loss and after a 6-month dietary intervention period from female continued weight losers (n 48) and weight regainers (n 48), evenly selected from four dietary groups that varied in protein and GI levels. The blood concentrations of twenty-nine proteins and three steroid hormones were measured. The changes in analytes during weight maintenance largely correlated negatively with the changes during weight loss, with some differences between continued weight losers and weight regainers. Increases in leptin (LEP) and C-reactive protein (CRP) were significantly associated with weight regain (P < 0·001 and P = 0·005, respectively), and these relationships were influenced by the diet. Consuming a high-protein and high-GI diet dissociated the positive relationship between the change in LEP concentration and weight regain. CRP increased during the weight-maintenance period only in weight regainers with a high-protein diet (P < 0·001). In addition, testosterone, luteinising hormone, angiotensinogen, plasminogen activator inhibitor-1, resistin, retinol-binding protein 4, insulin, glucagon, haptoglobin and growth hormone were also affected by the dietary intervention. The blood profile reflects not only the weight change during the maintenance period, but also the macronutrient composition of the dietary intervention, especially the protein level.

  10. Protein kinase M ζ and the maintenance of long-term memory.

    PubMed

    Zhang, Yang; Zong, Wei; Zhang, Lei; Ma, Yuanye; Wang, Jianhong

    2016-10-01

    Although various molecules have been found to mediate the processes of memory acquisition and consolidation, the molecular mechanism to maintain memory still remains elusive. In recent years, a molecular pathway focusing on protein kinase Mζ (PKMζ) has become of interest to researchers because of its potential role in long-term memory maintenance. PKMζ is an isoform of protein kinase C (PKC) and has a related structure that influences its function in maintaining memory. Considerable evidence has been gathered on PKMζ activity, including loss of function studies using PKMζ inhibitors, such as PKMζ inhibitory peptide (ZIP), suggesting PKMζ plays an important role in long-term memory maintenance. This review provides an overview of the role of PKMζ in long-term memory and outlines the molecular structure of PKMζ, the molecular mechanism of PKMζ in long-term memory maintenance and future directions of PKMζ research. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. A Light Harvesting Complex-Like Protein in Maintenance of Photosynthetic Components in Chlamydomonas.

    PubMed

    Zhao, Lei; Cheng, Dongmei; Huang, Xiahe; Chen, Mei; Dall'Osto, Luca; Xing, Jiale; Gao, Liyan; Li, Lingyu; Wang, Yale; Bassi, Roberto; Peng, Lianwei; Wang, Yingchun; Rochaix, Jean-David; Huang, Fang

    2017-08-01

    Using a genetic approach, we have identified and characterized a novel protein, named Msf1 (Maintenance factor for photosystem I), that is required for the maintenance of specific components of the photosynthetic apparatus in the green alga Chlamydomonas reinhardtii Msf1 belongs to the superfamily of light-harvesting complex proteins with three transmembrane domains and consensus chlorophyll-binding sites. Loss of Msf1 leads to reduced accumulation of photosystem I and chlorophyll-binding proteins/complexes. Msf1is a component of a thylakoid complex containing key enzymes of the tetrapyrrole biosynthetic pathway, thus revealing a possible link between Msf1 and chlorophyll biosynthesis. Protein interaction assays and greening experiments demonstrate that Msf1 interacts with Copper target homolog1 (CHL27B) and accumulates concomitantly with chlorophyll in Chlamydomonas, implying that chlorophyll stabilizes Msf1. Contrary to other light-harvesting complex-like genes, the expression of Msf1 is not stimulated by high-light stress, but its protein level increases significantly under heat shock, iron and copper limitation, as well as in stationary cells. Based on these results, we propose that Msf1 is required for the maintenance of photosystem I and specific protein-chlorophyll complexes especially under certain stress conditions. © 2017 American Society of Plant Biologists. All Rights Reserved.

  12. Role of Atypical Protein Kinases in Maintenance of Long-Term Memory and Synaptic Plasticity.

    PubMed

    Borodinova, A A; Zuzina, A B; Balaban, P M

    2017-03-01

    Investigation of biochemical mechanisms underlying the long-term storage of information in nervous system is one of main problems of modern neurobiology. As a molecular basis of long-term memory, long-term changes in kinase activities, increase in the level and changes in the subunit composition of receptors in synaptic membranes, local activity of prion-like proteins, and epigenetic modifications of chromatin have been proposed. Perhaps a combination of all or of some of these factors underlies the storage of long-term memory in the brain. Many recent studies have shown an exclusively important role of atypical protein kinases (PKCζ, PKMζ, and PKCι/λ) in processes of learning, consolidation and maintenance of memory. The present review is devoted to consideration of mechanisms of transcriptional and translational control of atypical protein kinases and their roles in induction and maintenance of long-term synaptic plasticity and memory in vertebrates and invertebrates.

  13. Separation of stem cell maintenance and transposon silencing functions of Piwi protein.

    PubMed

    Klenov, Mikhail S; Sokolova, Olesya A; Yakushev, Evgeny Y; Stolyarenko, Anastasia D; Mikhaleva, Elena A; Lavrov, Sergey A; Gvozdev, Vladimir A

    2011-11-15

    Piwi-interacting RNAs (piRNAs) and Piwi proteins have the evolutionarily conserved function of silencing of repetitive genetic elements in germ lines. The founder of the Piwi subfamily, Drosophila nuclear Piwi protein, was also shown to be required for the maintenance of germ-line stem cells (GSCs). Hence, null mutant piwi females exhibit two types of abnormalities, overexpression of transposons and severely underdeveloped ovaries. It remained unknown whether the failure of GSC maintenance is related to transposon derepression or if GSC self-renewal and piRNA silencing are two distinct functions of the Piwi protein. We have revealed a mutation, piwi(Nt), removing the nuclear localization signal of the Piwi protein. piwi(Nt) females retain the ability of GSC self-renewal and a near-normal number of egg chambers in the ovarioles but display a drastic transposable element derepression and nuclear accumulation of their transcripts in the germ line. piwi(Nt) mutants are sterile most likely because of the disturbance of piRNA-mediated transposon silencing. Analysis of chromatin modifications in the piwi(Nt) ovaries indicated that Piwi causes chromatin silencing only of certain types of transposons, whereas others are repressed in the nuclei without their chromatin modification. Thus, Piwi nuclear localization that is required for its silencing function is not essential for the maintenance of GSCs. We suggest that the Piwi function in GSC self-renewal is independent of transposon repression and is normally realized in the cytoplasm of GSC niche cells.

  14. Functional diversification of CLAVATA3-related CLE proteins in meristem maintenance in rice.

    PubMed

    Suzaki, Takuya; Yoshida, Akiko; Hirano, Hiro-Yuki

    2008-08-01

    Postembryonic development in plants depends on the activity of the shoot apical meristem (SAM) and root apical meristem (RAM). In Arabidopsis thaliana, CLAVATA signaling negatively regulates the size of the stem cell population in the SAM by repressing WUSCHEL. In other plants, however, studies of factors involved in stem cell maintenance are insufficient. Here, we report that two proteins closely related to CLAVATA3, FLORAL ORGAN NUMBER2 (FON2) and FON2-LIKE CLE PROTEIN1 (FCP1/Os CLE402), have functionally diversified to regulate the different types of meristem in rice (Oryza sativa). Unlike FON2, which regulates the maintenance of flower and inflorescence meristems, FCP1 appears to regulate the maintenance of the vegetative SAM and RAM. Constitutive expression of FCP1 results in consumption of the SAM in the vegetative phase, and application of an FCP1 CLE peptide in vitro disturbs root development by misspecification of cell fates in the RAM. FON1, a putative receptor of FON2, is likely to be unnecessary for these FCP1 functions. Furthermore, we identify a key amino acid residue that discriminates between the actions of FCP1 and FON2. Our results suggest that, although the basic framework of meristem maintenance is conserved in the angiosperms, the functions of the individual factors have diversified during evolution.

  15. The HU Protein Is Important for Apicoplast Genome Maintenance and Inheritance in Toxoplasma gondii

    PubMed Central

    Reiff, Sarah B.; Vaishnava, Shipra

    2012-01-01

    The apicoplast, a chloroplast-like organelle, is an essential cellular component of most apicomplexan parasites, including Plasmodium and Toxoplasma. The apicoplast maintains its own genome, a 35-kb DNA molecule that largely encodes proteins required for organellar transcription and translation. Interference with apicoplast genome maintenance and function is a validated target for drug therapy for malaria and toxoplasmosis. However, the many proteins required for genome maintenance and inheritance remain largely unstudied. Here we genetically characterize a nucleus-encoded homolog to the bacterial HU protein in Toxoplasma gondii. In bacteria, HU is a DNA-binding structural protein with fundamental roles in transcription, replication initiation, and DNA repair. Immunofluorescence assays reveal that in T. gondii this protein localizes to the apicoplast. We have found that the HU protein from Toxoplasma can successfully complement bacterial ΔhupA mutants, supporting a similar function. We were able to construct a genetic knockout of HU in Toxoplasma. This Δhu mutant is barely viable and exhibits significant growth retardation. Upon further analysis of the mutant phenotype, we find that this mutant has a dramatically reduced apicoplast genome copy number and, furthermore, suffers defects in the segregation of the apicoplast organelle. Our findings not only show that the HU protein is important for Toxoplasma cell biology but also demonstrate the importance of the apicoplast genome in the biogenesis of the organelle. PMID:22611021

  16. Noncanonical SMC protein in Mycobacterium smegmatis restricts maintenance of Mycobacterium fortuitum plasmids.

    PubMed

    Panas, Michael W; Jain, Paras; Yang, Hui; Mitra, Shimontini; Biswas, Debasis; Wattam, Alice Rebecca; Letvin, Norman L; Jacobs, William R

    2014-09-16

    Research on tuberculosis and leprosy was revolutionized by the development of a plasmid transformation system in the fast-growing surrogate, Mycobacterium smegmatis. This transformation system was made possible by the successful isolation of a M. smegmatis mutant strain mc(2)155, whose efficient plasmid transformation (ept) phenotype supported the replication of Mycobacterium fortuitum pAL5000 plasmids. In this report, we identified the EptC gene, the loss of which confers the ept phenotype. EptC shares significant amino acid sequence homology and domain structure with the MukB protein of Escherichia coli, a structural maintenance of chromosomes (SMC) protein. Surprisingly, M. smegmatis has three paralogs of SMC proteins: EptC and MSMEG_0370 both share homology with Gram-negative bacterial MukB; and MSMEG_2423 shares homology with Gram-positive bacterial SMCs, including the single SMC protein predicted for Mycobacterium tuberculosis and Mycobacterium leprae. Purified EptC was shown to bind ssDNA and stabilize negative supercoils in plasmid DNA. Moreover, an EptC-mCherry fusion protein was constructed and shown to bind to DNA in live mycobacteria, and to prevent segregation of plasmid DNA to daughter cells. To our knowledge, this is the first report of impaired plasmid maintenance caused by a SMC homolog, which has been canonically known to assist the segregation of genetic materials.

  17. Protein turnover forms one of the highest maintenance costs in Lactococcus lactis.

    PubMed

    Lahtvee, Petri-Jaan; Seiman, Andrus; Arike, Liisa; Adamberg, Kaarel; Vilu, Raivo

    2014-07-01

    Protein turnover plays an important role in cell metabolism by regulating metabolic fluxes. Furthermore, the energy costs for protein turnover have been estimated to account for up to a third of the total energy production during cell replication and hence may represent a major limiting factor in achieving either higher biomass or production yields. This work aimed to measure the specific growth rate (μ)-dependent abundance and turnover rate of individual proteins, estimate the ATP cost for protein production and turnover, and compare this with the total energy balance and other maintenance costs. The lactic acid bacteria model organism Lactococcus lactis was used to measure protein turnover rates at μ = 0.1 and 0.5 h(-1) in chemostat experiments. Individual turnover rates were measured for ~75% of the total proteome. On average, protein turnover increased by sevenfold with a fivefold increase in growth rate, whilst biomass yield increased by 35%. The median turnover rates found were higher than the specific growth rate of the bacterium, which suggests relatively high energy consumption for protein turnover. We found that protein turnover costs alone account for 38 and 47% of the total energy produced at μ = 0.1 and 0.5 h(-1), respectively, and gene ontology groups Energy metabolism and Translation dominated synthesis costs at both growth rates studied. These results reflect the complexity of metabolic changes that occur in response to changes in environmental conditions, and signify the trade-off between biomass yield and the need to produce ATP for maintenance processes.

  18. Role of cholecystokinin in induction and maintenance of dietary protein-stimulated pancreatic growth.

    PubMed

    Green, G M; Jurkowska, G; Berube, F L; Rivard, N; Guan, D; Morisset, J

    1992-04-01

    The role of cholecystokinin (CCK) in induction and maintenance of pancreatic growth stimulated by a high-protein diet was investigated. Rats adapted to 5% casein diet were switched to 70% casein for 21 days. MK-329, a CCK receptor antagonist, was administered at 2.5 mg.kg-1.day-1 ip, beginning on day zero (day zero treatment) or day 7 (midcourse treatment) of feeding 70% casein and thereafter. Another group was returned to 5% casein after 7 days of feeding 70% casein. Feeding 70% casein significantly stimulated increases of 32, 87, 74, 216, and 1,450% in pancreatic DNA, RNA, wet weight, protein content, and chymotrypsin content, respectively. Midcourse treatment with MK-329 was more effective than day zero treatment, and it completely reversed increases in pancreatic weight and RNA content, partially reversed increases in protein and chymotrypsin content, and had no effect on DNA content. Return to 5% casein rapidly reversed increases in pancreatic parameters, except for DNA. The results indicate that CCK is essential for induction and maintenance of dietary protein-stimulated pancreatic hypertrophy.

  19. Separation of stem cell maintenance and transposon silencing functions of Piwi protein

    PubMed Central

    Klenov, Mikhail S.; Sokolova, Olesya A.; Yakushev, Evgeny Y.; Stolyarenko, Anastasia D.; Mikhaleva, Elena A.; Lavrov, Sergey A.; Gvozdev, Vladimir A.

    2011-01-01

    Piwi-interacting RNAs (piRNAs) and Piwi proteins have the evolutionarily conserved function of silencing of repetitive genetic elements in germ lines. The founder of the Piwi subfamily, Drosophila nuclear Piwi protein, was also shown to be required for the maintenance of germ-line stem cells (GSCs). Hence, null mutant piwi females exhibit two types of abnormalities, overexpression of transposons and severely underdeveloped ovaries. It remained unknown whether the failure of GSC maintenance is related to transposon derepression or if GSC self-renewal and piRNA silencing are two distinct functions of the Piwi protein. We have revealed a mutation, piwiNt, removing the nuclear localization signal of the Piwi protein. piwiNt females retain the ability of GSC self-renewal and a near-normal number of egg chambers in the ovarioles but display a drastic transposable element derepression and nuclear accumulation of their transcripts in the germ line. piwiNt mutants are sterile most likely because of the disturbance of piRNA-mediated transposon silencing. Analysis of chromatin modifications in the piwiNt ovaries indicated that Piwi causes chromatin silencing only of certain types of transposons, whereas others are repressed in the nuclei without their chromatin modification. Thus, Piwi nuclear localization that is required for its silencing function is not essential for the maintenance of GSCs. We suggest that the Piwi function in GSC self-renewal is independent of transposon repression and is normally realized in the cytoplasm of GSC niche cells. PMID:22065765

  20. Yeast hnRNP-related proteins contribute to the maintenance of telomeres

    SciTech Connect

    Lee-Soety, Julia Y.; Jones, Jennifer; MacGibeny, Margaret A.; Remaly, Erin C.; Daniels, Lynsey; Ito, Andrea; Jean, Jessica; Radecki, Hannah; Spencer, Shannon

    2012-09-14

    Highlights: Black-Right-Pointing-Pointer Yeast hnRNP-related proteins are able to prevent faster senescence in telomerase-null cells. Black-Right-Pointing-Pointer The conserved RRMs in Npl3 are important for telomere maintenance. Black-Right-Pointing-Pointer Human hnRNP A1 is unable to complement the lack of NPL3 in yeast. Black-Right-Pointing-Pointer Npl3 and Cbc2 may work as telomere capping proteins. -- Abstract: Telomeres protect the ends of linear chromosomes, which if eroded to a critical length can become uncapped and lead to replicative senescence. Telomerase maintains telomere length in some cells, but inappropriate expression facilitates the immortality of cancer cells. Recently, proteins involved in RNA processing and ribosome assembly, such as hnRNP (heterogeneous nuclear ribonucleoprotein) A1, have been found to participate in telomere maintenance in mammals. The Saccharomyces cerevisiae protein Npl3 shares significant amino acid sequence similarities with hnRNP A1. We found that deleting NPL3 accelerated the senescence of telomerase null cells. The highly conserved RNA recognition motifs (RRM) in Npl3 appear to be important for preventing faster senescence. Npl3 preferentially binds telomere sequences in vitro, suggesting that Npl3 may affect telomeres directly. Despite similarities between the two proteins, human hnRNP A1 is unable to complement the lack of Npl3 to rescue accelerated senescence in tlc1 npl3 cells. Deletion of CBC2, which encodes another hnRNP-related protein that associates with Npl3, also accelerates senescence. Potential mechanisms by which hnRNP-related proteins maintain telomeres are discussed.

  1. Acute myeloid leukemia fusion proteins deregulate genes involved in stem cell maintenance and DNA repair

    PubMed Central

    Alcalay, Myriam; Meani, Natalia; Gelmetti, Vania; Fantozzi, Anna; Fagioli, Marta; Orleth, Annette; Riganelli, Daniela; Sebastiani, Carla; Cappelli, Enrico; Casciari, Cristina; Sciurpi, Maria Teresa; Mariano, Angela Rosa; Minardi, Simone Paolo; Luzi, Lucilla; Muller, Heiko; Di Fiore, Pier Paolo; Frosina, Guido; Pelicci, Pier Giuseppe

    2003-01-01

    Acute myelogenous leukemias (AMLs) are genetically heterogeneous and characterized by chromosomal rearrangements that produce fusion proteins with aberrant transcriptional regulatory activities. Expression of AML fusion proteins in transgenic mice increases the risk of myeloid leukemias, suggesting that they induce a preleukemic state. The underlying molecular and biological mechanisms are, however, unknown. To address this issue, we performed a systematic analysis of fusion protein transcriptional targets. We expressed AML1/ETO, PML/RAR, and PLZF/RAR in U937 hemopoietic precursor cells and measured global gene expression using oligonucleotide chips. We identified 1,555 genes regulated concordantly by at least two fusion proteins that were further validated in patient samples and finally classified according to available functional information. Strikingly, we found that AML fusion proteins induce genes involved in the maintenance of the stem cell phenotype and repress DNA repair genes, mainly of the base excision repair pathway. Functional studies confirmed that ectopic expression of fusion proteins constitutively activates pathways leading to increased stem cell renewal (e.g., the Jagged1/Notch pathway) and provokes accumulation of DNA damage. We propose that expansion of the stem cell compartment and induction of a mutator phenotype are relevant features underlying the leukemic potential of AML-associated fusion proteins. PMID:14660751

  2. A simple protein-energy wasting score predicts survival in maintenance hemodialysis patients.

    PubMed

    Moreau-Gaudry, Xavier; Jean, Guillaume; Genet, Leslie; Lataillade, Dominique; Legrand, Eric; Kuentz, François; Fouque, Denis

    2014-11-01

    Nutritional status is a powerful predictor of survival in maintenance hemodialysis patients but remains challenging to assess. We defined a new Protein Energy Wasting (PEW) score based on the nomenclature proposed by the International Society of Renal Nutrition and Metabolism in 2008. This score, graded from 0 (worse) to 4 (best) was derived from 4 body nutrition compartments: serum albumin, body mass index, a normalized serum creatinine value, and protein intake as assessed by nPNA. We applied this score to 1443 patients from the ARNOS prospective dialysis cohort and provide survival data from 2005 until 2008. Patients survival at 3.5 year. Survival ranged from 84%-69% according to the protein-energy wasting score. There was a clear-cut reduction in survival (5%-7%; P < 0.01) for each unit decrement in the score grade. There was a 99% survival at 1 year for patients with the score of 4. In addition, the 6-month variation of this PEW score also strongly predicted patients' survival (P < 0.01). A new simple and easy-to-get PEW score predicts survival in maintenance hemodialysis patients. Furthermore, increase of this nutritional score over time also indicates survival improvement, and may help to better identify subgroups of patients with a high mortality rate, in which nutrition support should be enforced. Copyright © 2014 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

  3. Potential role of minichromosome maintenance protein 2 as a screening biomarker in esophageal cancer high-risk population in China.

    PubMed

    Huang, Bo; Hu, Bin; Su, Min; Tian, Dongping; Guo, Yu; Lian, Shiyong; Liu, Zhicai; Wu, Xianying; Li, Qiaoshan; Zheng, Ruiming; Gao, Yuxia

    2011-06-01

    Minichromosome maintenance proteins are novel proliferative markers that have been proposed as diagnostic markers in many cancers. We evaluated the potential role of minichromosome maintenance protein 2 as a screening biomarker and compared it with proliferating cell nuclear antigen and Ki67 in a population survey of esophageal squamous cell carcinoma. A total of 299 esophageal samples from a high-risk region in China, including 171 from an endoscopy population survey, 30 from brushing cytology, and 98 from surgery and autopsy, underwent immunostaining with minichromosome maintenance protein 2, proliferating cell nuclear antigen, and Ki67 antibodies. Minichromosome maintenance protein 2 expression was confined to the proliferative compartment of normal and abnormal esophageal epithelium and particularly manifested in the surface layer of dysplasia and carcinoma in situ. The expression of proliferating cell nuclear antigen and Ki67 was positively correlated with that of minichromosome maintenance protein 2 (r(s) >0.39, P < .01); but their positive nuclei seldom reached the surface layer, and the labeling indices were significantly lower than those for minichromosome maintenance protein 2 in dysplasia (P < .05) and carcinoma in situ (P < .001). The sensitivity and specificity of minichromosome maintenance protein 2 in diagnosing dysplasia were 91.3% and 61.8%, respectively, higher than those for proliferating cell nuclear antigen (88.4% and 47.1%) and Ki67 (78.3% and 57.8%). Nine of 10 cancer and paracancerous surface-brushing samples expressed minichromosome maintenance protein 2, and the detection was higher than that for proliferating cell nuclear antigen (8/10 and 7/10) and Ki67 (7/10 and 7/10). However, none of 10 normal surface-brushing samples expressed the 3 markers. Minichromosome maintenance protein 2 is more sensitive and specific than proliferating cell nuclear antigen and Ki67 in indicating esophageal dysplasia. Minichromosome maintenance protein 2

  4. Expression, function, and targeting of the nuclear exporter chromosome region maintenance 1 (CRM1) protein.

    PubMed

    Ishizawa, Jo; Kojima, Kensuke; Hail, Numsen; Tabe, Yoko; Andreeff, Michael

    2015-09-01

    Nucleocytoplasmic trafficking of proteins/RNAs is essential to normal cellular function. Indeed, accumulating evidence suggests that cancer cells escape anti-neoplastic mechanisms and benefit from pro-survival signals via the dysregulation of this system. The nuclear exporter chromosome region maintenance 1 (CRM1) protein is the only protein in the karyopherin-β protein family that contributes to the trafficking of numerous proteins and RNAs from the nucleus. It is considered to be an oncogenic, anti-apoptotic protein in transformed cells, since it reportedly functions as a gatekeeper for cell survival, including affecting p53 function, and ribosomal biogenesis. Furthermore, abnormally high expression of CRM1 is correlated with poor patient prognosis in various malignancies. Therapeutic targeting of CRM1 has emerged as a novel cancer treatment strategy, starting with a clinical trial with leptomycin B, the original specific inhibitor of CRM1, followed by development of several next-generation small molecules. KPT-330, a novel member of the CRM1-selective inhibitors of nuclear export (SINE) class of compounds, is currently undergoing clinical evaluation for the therapy of various malignancies. Results from these trials suggest that SINE compounds may be particularly useful against hematological malignancies, which often become refractory to standard chemotherapeutic agents. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Expression, function, and targeting of the nuclear exporter chromosome region maintenance 1 (CRM1) protein

    PubMed Central

    Ishizawa, Jo; Kojima, Kensuke; Hail, Numsen; Tabe, Yoko; Andreeff, Michael

    2015-01-01

    Nucleocytoplasmic trafficking of proteins/RNAs is essential to normal cellular function. Indeed, accumulating evidence suggests that cancer cells escape anti-neoplastic mechanisms and benefit from pro-survival signals via the dysregulation of this system. The nuclear exporter chromosome region maintenance 1 (CRM1) protein is the only protein in the karyopherin-β protein family that contributes to the trafficking of numerous proteins and RNAs from the nucleus. It is considered to be an oncogenic, anti-apoptotic protein in transformed cells, since it reportedly functions as a gatekeeper for cell survival, including affecting p53 function, and ribosomal biogenesis. Furthermore, abnormally high expression of CRM1 is correlated with poor patient prognosis in various malignancies. Therapeutic targeting of CRM1 has emerged as a novel cancer treatment strategy, starting with a clinical trial with leptomycin B, the original specific inhibitor of CRM1, followed by development of several next-generation small molecules. KPT-330, a novel member of the CRM1-selective inhibitors of nuclear export (SINE) class of compounds, is currently undergoing clinical evaluation for the therapy of various malignancies. Results from these trials suggest that SINE compounds may be particularly useful against hematological malignancies, which often become refractory to standard chemotherapeutic agents. PMID:26048327

  6. High Dose Omega-3 Fatty Acid Administration and Skeletal Muscle Protein Turnover in Maintenance Hemodialysis Patients.

    PubMed

    Deger, Serpil Muge; Hung, Adriana M; Ellis, Charles D; Booker, Cindy; Bian, Aihua; Chen, Guanhua; Abumrad, Naji N; Ikizler, T Alp

    2016-07-07

    Protein energy wasting and systemic inflammation are prevalent in maintenance hemodialysis (MHD) patients. Omega-3 (ω-3) fatty acids have anti-inflammatory properties and have been shown to improve protein homeostasis. We hypothesized that administration of high-dose (2.9 g/d) ω-3 would be associated with decreased muscle protein breakdown in MHD patients with systemic inflammation. This is a substudy from a randomized, placebo-controlled study (NCT00655525). Patients were recruited between September 2008 and June 2011. Primary inclusion criteria included signs of chronic inflammation (average C-reactive protein of ≥5 mg/L over three consecutive measurements), lack of active infectious or inflammatory disease, no hospitalization within 1 month prior to the study, and not receiving steroids (>5 mg/d) and/or immunosuppressive agents. The primary outcomes were forearm muscle and whole body protein breakdown and synthesis before and after the intervention. The patients received ω-3 (n=11) versus placebo (n=9) for 12 weeks. Analysis of covariance was used to compare outcome variables at 12 weeks. Models were adjusted for a propensity score that was derived from age, sex, race, baseline high sensitivity C-reactive protein, diabetes mellitus, and fat mass because the groups were not balanced for several characteristics. Compared with placebo, ω-3 supplementation was significantly associated with decreased muscle protein breakdown at 12 weeks (-31, [interquartile range, -98--13] versus 26 [interquartile range, 13-87] µg/100 ml per min; P=0.01), which remained significant after multivariate adjustment (-46, [95% confidence interval, -102 to -1] µg/100 ml per min). ω-3 Supplementation resulted in decreased forearm muscle protein synthesis while the rate in the placebo group increased; however, there is no longer a statistically significant difference in skeletal muscle protein synthesis or in net protein balance after multivariate adjustment. There was no

  7. Diagnosis of pancreaticobiliary malignancy by detection of minichromosome maintenance protein 5 in biliary brush cytology

    PubMed Central

    Keane, Margaret G; Huggett, Matthew T; Chapman, Michael H; Johnson, Gavin J; Webster, George J; Thorburn, Douglas; Mackay, James; Pereira, Stephen P

    2017-01-01

    Background: Biliary brush cytology is the standard method of evaluating biliary strictures, but is insensitive at detecting malignancy. In pancreaticobiliary cancer minichromosome maintenance replication proteins (MCM 2–7) are dysregulated in the biliary epithelium and MCM5 levels are elevated in bile samples. This study aimed to validate an immunocolorimetric ELISA assay for MCM5 as a pancreaticobiliary cancer biomarker in biliary brush samples. Methods: Biliary brush specimens were collected prospectively at ERCP from patients with a biliary stricture. Collected samples were frozen at −80 °C. The supernatant was washed and lysed cells incubated with HRP-labelled anti-MCM5 mouse monoclonal antibody. Test positivity was determined by optical density absorbance. Patients underwent biliary brush cytology or additional investigations as per clinical routine. Results: Ninety-seven patients were included in the study; 50 had malignant strictures. Median age was 65 years (range 21–94) and 51 were male. Compared with final diagnosis the MCM5 assay had a sensitivity for malignancy of 65.4% compared with 25.0% for cytology. In the 72 patients with paired MCM5 assay and biliary brush cytology, MCM5 demonstrated an improved sensitivity (55.6% vs 25.0% P=0.0002) for the detection of malignancy. Conclusions: Minichromosome maintenance replication protein5 is a more sensitive indicator of pancreaticobiliary malignancy than standard biliary brush cytology. PMID:28081547

  8. BDNF Facilitates L-LTP Maintenance in the Absence of Protein Synthesis through PKMζ

    PubMed Central

    Mei, Fan; Nagappan, Guhan; Ke, Yang; Sacktor, Todd C.; Lu, Bai

    2011-01-01

    Late-phase long term potentiation (L-LTP) is thought to be the cellular basis for long-term memory (LTM). While LTM as well as L-LTP is known to depend on transcription and translation, it is unclear why brain-derived neurotrophic factor (BDNF) could sustain L-LTP when protein synthesis is inhibited. The persistently active protein kinase ζ (PKMζ) is the only molecule implicated in perpetuating L-LTP maintenance. Here, in mouse acute brain slices, we show that inhibition of PKMζ reversed BDNF-dependent form of L-LTP. While BDNF did not alter the steady-state level of PKMζ, BDNF together with the L-LTP inducing theta-burst stimulation (TBS) increased PKMζ level even without protein synthesis. Finally, in the absence of de novo protein synthesis, BDNF maintained TBS-induced PKMζ at a sufficient level. These results suggest that BDNF sustains L-LTP through PKMζ in a protein synthesis-independent manner, revealing an unexpected link between BDNF and PKMζ. PMID:21747912

  9. Bromodomain Proteins Contribute to Maintenance of Bloodstream Form Stage Identity in the African Trypanosome

    PubMed Central

    Schulz, Danae; Mugnier, Monica R.; Paulsen, Eda-Margaret; Kim, Hee-Sook; Chung, Chun-wa W.; Tough, David F.; Rioja, Inmaculada; Prinjha, Rab K.; Papavasiliou, F. Nina; Debler, Erik W.

    2015-01-01

    Trypanosoma brucei, the causative agent of African sleeping sickness, is transmitted to its mammalian host by the tsetse. In the fly, the parasite’s surface is covered with invariant procyclin, while in the mammal it resides extracellularly in its bloodstream form (BF) and is densely covered with highly immunogenic Variant Surface Glycoprotein (VSG). In the BF, the parasite varies this highly immunogenic surface VSG using a repertoire of ~2500 distinct VSG genes. Recent reports in mammalian systems point to a role for histone acetyl-lysine recognizing bromodomain proteins in the maintenance of stem cell fate, leading us to hypothesize that bromodomain proteins may maintain the BF cell fate in trypanosomes. Using small-molecule inhibitors and genetic mutants for individual bromodomain proteins, we performed RNA-seq experiments that revealed changes in the transcriptome similar to those seen in cells differentiating from the BF to the insect stage. This was recapitulated at the protein level by the appearance of insect-stage proteins on the cell surface. Furthermore, bromodomain inhibition disrupts two major BF-specific immune evasion mechanisms that trypanosomes harness to evade mammalian host antibody responses. First, monoallelic expression of the antigenically varied VSG is disrupted. Second, rapid internalization of antibodies bound to VSG on the surface of the trypanosome is blocked. Thus, our studies reveal a role for trypanosome bromodomain proteins in maintaining bloodstream stage identity and immune evasion. Importantly, bromodomain inhibition leads to a decrease in virulence in a mouse model of infection, establishing these proteins as potential therapeutic drug targets for trypanosomiasis. Our 1.25Å resolution crystal structure of a trypanosome bromodomain in complex with I-BET151 reveals a novel binding mode of the inhibitor, which serves as a promising starting point for rational drug design. PMID:26646171

  10. Intercellular chaperone transmission via exosomes contributes to maintenance of protein homeostasis at the organismal level.

    PubMed

    Takeuchi, Toshihide; Suzuki, Mari; Fujikake, Nobuhiro; Popiel, H Akiko; Kikuchi, Hisae; Futaki, Shiroh; Wada, Keiji; Nagai, Yoshitaka

    2015-05-12

    The heat shock response (HSR), a transcriptional response that up-regulates molecular chaperones upon heat shock, is necessary for cell survival in a stressful environment to maintain protein homeostasis (proteostasis). However, there is accumulating evidence that the HSR does not ubiquitously occur under stress conditions, but largely depends on the cell types. Despite such imbalanced HSR among different cells and tissues, molecular mechanisms by which multicellular organisms maintain their global proteostasis have remained poorly understood. Here, we report that proteostasis can be maintained by molecular chaperones not only in a cell-autonomous manner but also in a non-cell-autonomous manner. We found that elevated expression of molecular chaperones, such as Hsp40 and Hsp70, in a group of cells improves proteostasis in other groups of cells, both in cultured cells and in Drosophila expressing aggregation-prone polyglutamine proteins. We also found that Hsp40, as well as Hsp70 and Hsp90, is physiologically secreted from cells via exosomes, and that the J domain at the N terminus is responsible for its exosome-mediated secretion. Addition of Hsp40/Hsp70-containing exosomes to the culture medium of the polyglutamine-expressing cells results in efficient suppression of inclusion body formation, indicating that molecular chaperones non-cell autonomously improve the protein-folding environment via exosome-mediated transmission. Our study reveals that intercellular chaperone transmission mediated by exosomes is a novel molecular mechanism for non-cell-autonomous maintenance of organismal proteostasis that could functionally compensate for the imbalanced state of the HSR among different cells, and also provides a novel physiological role of exosomes that contributes to maintenance of organismal proteostasis.

  11. Acetylation Reader Proteins: Linking Acetylation Signaling to Genome Maintenance and Cancer.

    PubMed

    Gong, Fade; Chiu, Li-Ya; Miller, Kyle M

    2016-09-01

    Chromatin-based DNA damage response (DDR) pathways are fundamental for preventing genome and epigenome instability, which are prevalent in cancer. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) catalyze the addition and removal of acetyl groups on lysine residues, a post-translational modification important for the DDR. Acetylation can alter chromatin structure as well as function by providing binding signals for reader proteins containing acetyl-lysine recognition domains, including the bromodomain (BRD). Acetylation dynamics occur upon DNA damage in part to regulate chromatin and BRD protein interactions that mediate key DDR activities. In cancer, DDR and acetylation pathways are often mutated or abnormally expressed. DNA damaging agents and drugs targeting epigenetic regulators, including HATs, HDACs, and BRD proteins, are used or are being developed to treat cancer. Here, we discuss how histone acetylation pathways, with a focus on acetylation reader proteins, promote genome stability and the DDR. We analyze how acetylation signaling impacts the DDR in the context of cancer and its treatments. Understanding the relationship between epigenetic regulators, the DDR, and chromatin is integral for obtaining a mechanistic understanding of genome and epigenome maintenance pathways, information that can be leveraged for targeting acetylation signaling, and/or the DDR to treat diseases, including cancer.

  12. Intimin-Mediated Export of Passenger Proteins Requires Maintenance of a Translocation-Competent Conformation

    PubMed Central

    Adams, Thorsten M.; Wentzel, Alexander; Kolmar, Harald

    2005-01-01

    Intimins from pathogenic bacteria promote intimate bacterial adhesion to epithelial cells. Several structurally similar domains form on the bacterial cell surface an extended rigid rod that exposes the carboxy-terminal domain, which interacts with the translocated intimin receptor. We constructed a series of intimin-derived fusion proteins consisting of carboxy-terminally truncated intimin and the immunoglobulin light-chain variable domain REIv, ubiquitin, calmodulin, β-lactamase inhibitor protein, or β-lactamase. By systematically investigating the intimin-mediated cell surface exposure of these passenger domains in the presence or absence of compounds that interfere with outer membrane stability or passenger domain folding, we acquired experimental evidence that intimin-mediated protein export across the outer membrane requires, prior to export, the maintenance of a translocation-competent conformation that may be distinct from the final protein structure. We propose that, during export, competition exists between productive translocation and folding of the passenger domain in the periplasm into a stable conformation that is not compatible with translocation through the bacterial outer membrane. These results may expand understanding of the mechanism by which intimins are inserted into the outer membrane and expose extracellular domains on the cell surface. PMID:15629924

  13. Small-molecule tools for dissecting the roles of SSB/protein interactions in genome maintenance

    SciTech Connect

    Lu, Duo; Bernstein, Douglas A.; Satyshur, Kenneth A.; Keck, James L.

    2010-09-03

    Bacterial single-stranded DNA-binding proteins (SSBs) help to recruit a diverse array of genome maintenance enzymes to their sites of action through direct protein interactions. For all cases examined to date, these interactions are mediated by the evolutionarily conserved C terminus of SSB (SSB-Ct). The essential nature of SSB protein interactions makes inhibitors that block SSB complex formation valuable biochemical tools and attractive potential antibacterial agents. Here, we identify four small molecules that disrupt complexes formed between Escherichia coli SSB and Exonuclease I (ExoI), a well-studied SSB-interacting enzyme. Each compound disrupts ExoI/SSB-Ct peptide complexes and abrogates SSB stimulation of ExoI nuclease activity. Structural and biochemical studies support a model for three of the compounds in which they compete with SSB for binding to ExoI. The fourth appears to rely on an allosteric mechanism to disrupt ExoI/SSB complexes. Subsets of the inhibitors block SSB-Ct complex formation with two other SSB-interaction partners as well, which highlights their utility as reagents for investigating the roles of SSB/protein interactions in diverse DNA replication, recombination, and repair reactions.

  14. Acetylation Reader Proteins: Linking Acetylation Signaling to Genome Maintenance and Cancer

    PubMed Central

    Miller, Kyle M.

    2016-01-01

    Chromatin-based DNA damage response (DDR) pathways are fundamental for preventing genome and epigenome instability, which are prevalent in cancer. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) catalyze the addition and removal of acetyl groups on lysine residues, a post-translational modification important for the DDR. Acetylation can alter chromatin structure as well as function by providing binding signals for reader proteins containing acetyl-lysine recognition domains, including the bromodomain (BRD). Acetylation dynamics occur upon DNA damage in part to regulate chromatin and BRD protein interactions that mediate key DDR activities. In cancer, DDR and acetylation pathways are often mutated or abnormally expressed. DNA damaging agents and drugs targeting epigenetic regulators, including HATs, HDACs, and BRD proteins, are used or are being developed to treat cancer. Here, we discuss how histone acetylation pathways, with a focus on acetylation reader proteins, promote genome stability and the DDR. We analyze how acetylation signaling impacts the DDR in the context of cancer and its treatments. Understanding the relationship between epigenetic regulators, the DDR, and chromatin is integral for obtaining a mechanistic understanding of genome and epigenome maintenance pathways, information that can be leveraged for targeting acetylation signaling, and/or the DDR to treat diseases, including cancer. PMID:27631103

  15. Human CDC45 protein binds to minichromosome maintenance 7 protein and the p70 subunit of DNA polymerase alpha.

    PubMed

    Kukimoto, I; Igaki, H; Kanda, T

    1999-11-01

    Budding yeast CDC45 encodes Cdc45p, an essential protein required to trigger initiation of DNA replication in late G1 phase. We cloned four and one species of the human Cdc45p homolog cDNA, resulting from different splicing patterns, from HeLa cell and human placenta cDNA libraries, respectively. A comparison of the cDNAs and the genomic sequence showed that the longest encoding a 610-amino acid protein was comprised of 20 exons. One species, which lacks exon 7 and contains the shorter of two exons 18, was identical with the previously reported CDC45L cDNA and constituted 24 out of 28 clones from HeLa cells. Splicing was different in HeLa cells and TIG-1 cells, a human diploid cell line. Human CDC45 protein was found to bind directly in vitro to human minichromosome maintenance 7 protein (hMCM7) and to the p70 subunit of DNA polymerase alpha. The data support a thesis that human CDC45 acts as a molecular tether to mediate loading of the DNA polymerase alpha on to the DNA replication complex through binding to hMCM7.

  16. Role of platelets in maintenance of pulmonary vascular permeability to protein

    SciTech Connect

    Lo, S.K.; Burhop, K.E.; Kaplan, J.E.; Malik, A.B. )

    1988-04-01

    The authors examined the role of platelets in maintenance of pulmonary vascular integrity by inducing thrombocytopenia in sheep using antiplatelet serum (APS). A causal relationship between thrombocytopenia and increase in pulmonary vascular permeability was established by platelet repletion using platelet-rich plasma (PRP). Sheep were chronically instrumented and lung lymph fistulas prepared to monitor pulmonary lymph flow (Q{sub lym}). A balloon catheter was positioned in the left atrium to assess pulmonary vascular permeability to protein after raising the left atrial pressure (P{sub la}). Thrombocytopenia was maintained for 3 days by daily intramuscular APS injections. In studies using cultured bovine pulmonary artery endothelial monolayers, transendothelia permeability of {sup 125}I-labeled albumin was reduced 50 and 95%, respectively, when 2.5 {times} 10{sup 7} or 5 {times} 10{sup 7} platelets were added onto endothelial monolayers. However, addition of 5 {times} 10{sup 6} platelets or 5 {times} 10{sup 7} red blood cells did not reduce endothelial monolayer albumin permeability. Results indicate that platelets are required for the maintenance of pulmonary vascular permeability. Reduction in permeability appears to involve an interaction of platelets with the endothelium.

  17. Structure of an octameric form of the minichromosome maintenance protein from the archaeon Pyrococcus abyssi

    PubMed Central

    Cannone, Giuseppe; Visentin, Silvia; Palud, Adeline; Henneke, Ghislaine; Spagnolo, Laura

    2017-01-01

    Cell division is a complex process that requires precise duplication of genetic material. Duplication is concerted by replisomes. The Minichromosome Maintenance (MCM) replicative helicase is a crucial component of replisomes. Eukaryotic and archaeal MCM proteins are highly conserved. In fact, archaeal MCMs are powerful tools for elucidating essential features of MCM function. However, while eukaryotic MCM2-7 is a heterocomplex made of different polypeptide chains, the MCM complexes of many Archaea form homohexamers from a single gene product. Moreover, some archaeal MCMs are polymorphic, and both hexameric and heptameric architectures have been reported for the same polypeptide. Here, we present the structure of the archaeal MCM helicase from Pyrococcus abyssi in its single octameric ring assembly. To our knowledge, this is the first report of a full-length octameric MCM helicase. PMID:28176822

  18. Diets with High or Low Protein Content and Glycemic Index for Weight-Loss Maintenance

    PubMed Central

    Larsen, Thomas Meinert; Dalskov, Stine-Mathilde; van Baak, Marleen; Jebb, Susan A.; Papadaki, Angeliki; Pfeiffer, Andreas F.H.; Martinez, J. Alfredo; Handjieva-Darlenska, Teodora; Kunešová, Marie; Pihlsgård, Mats; Stender, Steen; Holst, Claus; Saris, Wim H.M.; Astrup, Arne

    2012-01-01

    Background Studies of weight-control diets that are high in protein or low in glycemic index have reached varied conclusions, probably owing to the fact that the studies had insufficient power. Methods We enrolled overweight adults from eight European countries who had lost at least 8% of their initial body weight with a 3.3-MJ (800-kcal) low-calorie diet. Participants were randomly assigned, in a two-by-two factorial design, to one of five ad libitum diets to prevent weight regain over a 26-week period: a low-protein and low-glycemic-index diet, a low-protein and high-glycemic-index diet, a high-protein and low-glycemic-index diet, a high-protein and high-glycemic-index diet, or a control diet. Results A total of 1209 adults were screened (mean age, 41 years; body-mass index [the weight in kilograms divided by the square of the height in meters], 34), of whom 938 entered the low-calorie-diet phase of the study. A total of 773 participants who completed that phase were randomly assigned to one of the five maintenance diets; 548 completed the intervention (71%). Fewer participants in the high-protein and the low-glycemic-index groups than in the low-protein–high-glycemic-index group dropped out of the study (26.4% and 25.6%, respectively, vs. 37.4%; P = 0.02 and P = 0.01 for the respective comparisons). The mean initial weight loss with the low-calorie diet was 11.0 kg. In the analysis of participants who completed the study, only the low-protein–high-glycemic-index diet was associated with subsequent significant weight regain (1.67 kg; 95% confidence interval [CI], 0.48 to 2.87). In an intention-to-treat analysis, the weight regain was 0.93 kg less (95% CI, 0.31 to 1.55) in the groups assigned to a high-protein diet than in those assigned to a low-protein diet (P = 0.003) and 0.95 kg less (95% CI, 0.33 to 1.57) in the groups assigned to a low-glycemic-index diet than in those assigned to a high-glycemic-index diet (P = 0.003). The analysis involving

  19. Effect of Ramipril on Urinary Protein Excretion in Maintenance Renal Transplant Patients Converted to Sirolimus.

    PubMed

    Mandelbrot, D A; Alberú, J; Barama, A; Marder, B A; Silva, H T; Flechner, S M; Flynn, A; Healy, C; Li, H; Tortorici, M A; Schulman, S L

    2015-12-01

    This prospective, randomized, double-blind, placebo-controlled study evaluated the effects of ramipril on urinary protein excretion in renal transplant patients treated with sirolimus following conversion from a calcineurin inhibitor. Patients received ramipril or placebo for up to 6 weeks before conversion and 52 weeks thereafter. Doses were increased if patients developed proteinuria (urinary protein/creatinine ratio ≥0.5); losartan was given as rescue therapy for persistent proteinuria. The primary end point was time to losartan initiation. Of 295 patients randomized, 264 met the criteria for sirolimus conversion (ramipril, 138; placebo, 126). At 52 weeks, the cumulative rate of losartan initiation was significantly lower with ramipril (6.2%) versus placebo (23.2%) (p < 0.001). No significant differences were observed between ramipril and placebo for change in glomerular filtration rate from baseline (p = 0.148) or in the number of patients with biopsy-confirmed acute rejection (13 vs. 5, respectively; p = 0.073). One patient in the placebo group died due to cerebrovascular accident. Treatment-emergent adverse events were consistent with the known safety profile of sirolimus and were not potentiated by ramipril co-administration. Ramipril was effective in reducing the incidence of proteinuria for up to 1 year following conversion to sirolimus in maintenance renal transplant patients.

  20. Selective inhibition of protein arginine methyltransferase 5 blocks initiation and maintenance of B-cell transformation

    PubMed Central

    Alinari, Lapo; Mahasenan, Kiran V.; Yan, Fengting; Karkhanis, Vrajesh; Chung, Ji-Hyun; Smith, Emily M.; Quinion, Carl; Smith, Porsha L.; Kim, Lisa; Patton, John T.; Lapalombella, Rosa; Yu, Bo; Wu, Yun; Roy, Satavisha; De Leo, Alessandra; Pileri, Stefano; Agostinelli, Claudio; Ayers, Leona; Bradner, James E.; Chen-Kiang, Selina; Elemento, Olivier; Motiwala, Tasneem; Majumder, Sarmila; Byrd, John C.; Jacob, Samson; Sif, Said; Li, Chenglong

    2015-01-01

    Epigenetic events that are essential drivers of lymphocyte transformation remain incompletely characterized. We used models of Epstein-Barr virus (EBV)–induced B-cell transformation to document the relevance of protein arginine methyltransferase 5 (PRMT5) to regulation of epigenetic-repressive marks during lymphomagenesis. EBV+ lymphomas and transformed cell lines exhibited abundant expression of PRMT5, a type II PRMT enzyme that promotes transcriptional silencing of target genes by methylating arginine residues on histone tails. PRMT5 expression was limited to EBV-transformed cells, not resting or activated B lymphocytes, validating it as an ideal therapeutic target. We developed a first-in-class, small-molecule PRMT5 inhibitor that blocked EBV-driven B-lymphocyte transformation and survival while leaving normal B cells unaffected. Inhibition of PRMT5 led to lost recruitment of a PRMT5/p65/HDAC3-repressive complex on the miR96 promoter, restored miR96 expression, and PRMT5 downregulation. RNA-sequencing and chromatin immunoprecipitation experiments identified several tumor suppressor genes, including the protein tyrosine phosphatase gene PTPROt, which became silenced during EBV-driven B-cell transformation. Enhanced PTPROt expression following PRMT5 inhibition led to dephosphorylation of kinases that regulate B-cell receptor signaling. We conclude that PRMT5 is critical to EBV-driven B-cell transformation and maintenance of the malignant phenotype, and that PRMT5 inhibition shows promise as a novel therapeutic approach for B-cell lymphomas. PMID:25742700

  1. Cerebral klotho protein as a humoral factor for maintenance of baroreflex.

    PubMed

    Chen, L-J; Cheng, M-F; Ku, P-M; Cheng, J-T

    2015-02-01

    The klotho protein produced by the choroid plexus is known as a humoral factor in central nervous system. Many hormones affecting the baroreflex sensitivity have been introduced in the brain. However, role of klotho in the baroreflex sensitivity is still unknown. Recently, mutations in the klotho gene have been linked to cardiovascular diseases in both animals and human subjects. Also, silencing of brain klotho has been reported to enhance cold-induced elevation of blood pressure. Thus, we investigated the role of klotho in maintenance of central cardiovascular reflex sensitivity. Male Wistar Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs) were used. Either klotho shRNA or scramble shRNA was also ICV-infused into the brains of WKY rats to investigate the role of klotho in brain. Recombinant klotho or rat IgG was infused into the cerebral paraventricle (ICV) of SHRs for further understanding the role of klotho in hypertension. The baroreflex sensitivity was detected using the challenge with a depressor dose of sodium nitroprusside (SNP, 50 μg/kg) or with a pressor dose of phenylephrine (PE, 8 μg/kg). We found that silencing of klotho expression in the brain decreased the baroreflex sensitivity in WKY rats. Also, modulation of the blood pressure for one week altered the cardiovascular homeostasis and resulted in an increased expression of klotho in medulla oblongata. Moreover, the baroreflex sensitivity was restored in SHRs that received recombinant klotho through ICV brain. Thus, klotho is involved in the maintenance of baroreflex sensitivity in the brain.

  2. Maintenance of pluripotency in human embryonic stem cells stably over-expressing enhanced green fluorescent protein.

    PubMed

    Liu, Yi-Ping; Dovzhenko, Oksana V; Garthwaite, Mark A; Dambaeva, Svetlana V; Durning, Maureen; Pollastrini, Leah M; Golos, Thaddeus G

    2004-12-01

    The availability of human embryonic stem (HES) cells with a readily evaluated genetic marker such as green fluorescent protein (GFP) could facilitate a number of experimental opportunities. We constructed a novel plasmid with two elongation factor-1alpha (EF-1alpha) promoters (YPL2) to obtain a vector with mammalian promoters for simultaneous transgene expression in HES cells. An enhanced green fluorescent protein (EGFP) cDNA was inserted under the control of the first EF-1alpha promoter to construct plasmid YPL2-EGFP. The second EF1-alpha promoter was upstream of the neomycin resistance gene. H1 HES cells were transfected with YPL2-EGFP using Fugene 6. Following 100 microg/ml neomycin selection, individual colonies demonstrating stable EGFP expression were observed. After 4 months of passage under neomycin selection, the cells continued to maintain typical HES cell morphology. Undifferentiated cells showed no change in EGFP expression as determined by FACS analysis. Immunostaining demonstrated maintenance of Oct-3/4 expression in undifferentiated H1EGFP cells that was indistinguishable from wild-type HES cells. Addition of 10 ng/ml bone morphogenic protein-4 (BMP-4) to the cells provoked morphological and functional differentiation to trophoblasts, but no loss of EGFP expression. Following injection of EGFP-HES cells into immunodeficient mice, there was robust formation of teratomas that demonstrated a broad range of morphological pluripotency with widespread EGFP expression. EGFP expression was also maintained in differentiating embryoid bodies formed from EGFP-HES cells. This report demonstrates that ES cells carrying EGFP will be useful in diverse areas of embryonic stem cell research.

  3. Minichromosome maintenance protein 7 regulates phagocytosis in kuruma shrimp Marsupenaeus japonicas against white spot syndrome virus.

    PubMed

    Wang, Zhi; Zhu, Fei

    2016-08-01

    Minichromosome maintenance protein (MCM7) belongs to the MCM protein family and participates in the MCM complex by playing a role in the cell replication cycle and chromosome initiation of eukaryotes. Previously, we found that several genes, including MCM7, were over-expressed in Drosophila melanogaster after white spot syndrome virus (WSSV) infection. In this study, we aimed to further research the MCM7 of kuruma shrimp, Marsupenaeus japonicus (mjMCM7) and determine its role in the innate immune system. To this end, we cloned the entire 2307-bp mjMCM7 sequence, including a 1974-bp open reading frame (ORF) encoding a 658-aa-long protein. Real-time PCR showed that the gene was primarily expressed in the hemolymph and hepatopancreas and over-expressed in shrimp challenged with WSSV. Gene function study was carried out by knocking down the expression of MCM7 using small interference RNA (siRNA). The results revealed that β-actin, hemocyanin, prophenoloxidase (proPO) and tumor necrosis factor-α (TNF-α) were up-regulated while the cytoskeleton proteins such as myosin and Rho were significantly down-regulated at 24 h after treatment. The results indicate a possible relationship between mjMCM7 and the innate immune system, and suggest that mjMCM7 may play a role in phagocytosis. After WSSV challenge, WSSV copies and mortality count were both higher in the MCM7-siRNA-treated groups at 60 h after treatment, and the mortality count approached that of the control groups over time. The phagocytosis rate was significantly lower in the MCM7-siRNA-treated group than in the WSSV group. The findings of this study confirm that mjMCM7 positively regulates phagocytosis and plays an important role against WSSV. These results could help researchers to further understand the function of the MCM7 protein and reveal its potential role in the innate immunity of invertebrates.

  4. British Dietetic Association evidence-based guidelines for the protein requirements of adults undergoing maintenance haemodialysis or peritoneal dialysis.

    PubMed

    Naylor, H L; Jackson, H; Walker, G H; Macafee, S; Magee, K; Hooper, L; Stewart, L; MacLaughlin, H L

    2013-08-01

    Existing nutritional guidelines suggest that protein requirements of adults with stage five chronic kidney disease undergoing haemodialysis (HD) or peritoneal dialysis (PD) are increased as a result of protein losses during dialysis. The present review aimed to update previous guidance and develop evidence-based practice guidelines on the protein requirements of adults undergoing maintenance dialysis. Following a PICO approach (Participants or Population, Intervention or Exposure, Comparison and Outcome), four research questions were formulated to investigate the total protein requirement and protein quality required by adults undergoing HD and PD. A comprehensive, systematic review was undertaken using the databases Medline, EMBASE and the Cochrane Library from 2005 to September 2009 for HD studies and from 1997 to September 2009 for PD studies. The literature search yielded 2931 studies, which were assessed for inclusion. Following appraisal, 19 studies in HD and 18 studies in PD met the inclusion criteria and were systematically reviewed. Limited good quality evidence supports the recommendations that: (i) adults undergoing maintenance HD require a minimum protein intake of 1.1 g kg(-1) ideal body weight (IBW) per day; and (ii) adults undergoing maintenance PD require a minimum protein intake of 1.0-1.2 kg(-1) IBW per day, in conjunction with an adequate energy intake. There were no studies that addressed the quality of protein for either HD or PD. Evidence suggests that nutritional status may be maintained with lower protein intakes than previously recommended. However, the evidence base is limited and further randomised controlled trials are required to establish the optimal protein intake for dialysis patients. © 2013 The Authors Journal of Human Nutrition and Dietetics © 2013 The British Dietetic Association Ltd.

  5. The kinesin related motor protein, Eg5, is essential for maintenance of pre-implantation embryogenesis

    SciTech Connect

    Castillo, Andrew; Justice, Monica J. . E-mail: mjustice@bcm.tmc.edu

    2007-06-08

    Eg5 is a plus end directed kinesin related motor protein (KRP) previously shown to be involved in the assembly and maintenance of the mitotic spindle. KRPs are molecular motors capable of generating forces upon microtubules (MTs) in dividing cells and driving structural rearrangements necessary in the developing spindle. In vitro experiments demonstrate that loss of Eg5 results in cell cycle arrest and defective centrosome separation resulting in the development of monopolar spindles. Here we describe mice with a genetrap insertion in Eg5. Heterozygous mutant mice appear phenotypically normal. In contrast, embryos homozygous for the Eg5 null allele recovered at embryonic days 2.5-3.5 display signs of a proliferation defect as reduced cell numbers and failure of compaction and progression to the blastocyst stage was observed. These data, in conjunction with previous in vitro data, suggest that loss of Eg5 results in abnormal spindle structure, cell cycle arrest and thereby reduced cell proliferation of early cleavage pre-implantation embryos. These observations further support the conclusion that Eg5 is essential for cell division early in mouse development, and that maternal contribution may sustain the embryo through the maternal to zygotic transition at which point supplies of functional Eg5 are exhausted, preventing further cell cleavage.

  6. Checkpoint proteins influence telomeric silencing and length maintenance in budding yeast.

    PubMed Central

    Longhese, M P; Paciotti, V; Neecke, H; Lucchini, G

    2000-01-01

    A complex network of surveillance mechanisms, called checkpoints, interrupts cell cycle progression when damage to the genome is detected or when cells fail to complete DNA replication, thus ensuring genetic integrity. In budding yeast, components of the DNA damage checkpoint regulatory network include the RAD9, RAD17, RAD24, MEC3, DDC1, RAD53, and MEC1 genes that are proposed to be involved in different aspects of DNA metabolism. We provide evidence that some DNA damage checkpoint components play a role in maintaining telomere integrity. In fact, rad53 mutants specifically enhance repression of telomere-proximal transcription via the Sir-mediated pathway, suggesting that Rad53 might be required for proper chromatin structure at telomeres. Moreover, Rad53, Mec1, Ddc1, and Rad17 are necessary for telomere length maintenance, since mutations in all of these genes cause a decrease in telomere size. The telomeric shortening in rad53 and mec1 mutants is further enhanced in the absence of SIR genes, suggesting that Rad53/Mec1 and Sir proteins contribute to chromosome end protection by different pathways. The finding that telomere shortening, but not increased telomeric repression of gene expression in rad53 mutants, can be suppressed by increasing dNTP synthetic capacity in these strains suggests that transcriptional silencing and telomere integrity involve separable functions of Rad53. PMID:10924458

  7. Maintenance of Membrane Integrity and Permeability Depends on a Patched-Related Protein in Caenorhabditis elegans.

    PubMed

    Choi, Myung-Kyu; Son, Sangwon; Hong, Mingi; Choi, Min Sung; Kwon, Jae Young; Lee, Junho

    2016-04-01

    Membrane integrity is critical for cell survival, defects of which cause pathological symptoms such as metabolic diseases. In this study, we used ethanol sensitivity of the nematode Caenorhabditis elegans to identify genetic factors involved in membrane integrity. InC. elegans, acute exposure to a high concentration (7% v/v) of ethanol changes membrane permeability, as measured by propidium iodide staining, and causes paralysis. We used the timing of complete paralysis as an indicator for alteration of membrane integrity in our genetic screen, and identified ptr-6 as a gene that confers ethanol resistance when mutated. PTR-6 is a patched-related protein and contains a sterol sensing domain. Inhibition of two PTR-encoding genes,ptr-15 and ptr-23, and mboa-1, encoding an Acyl Co-A: cholesterol acyltransferase homolog, restored ethanol sensitivity of the ptr-6 mutant, suggesting that these ptr genes and mboa-1 are involved in the maintenance of membrane integrity and permeability. Our results suggest that C. elegans can be used as a model system to identify factors involved in metabolic diseases and to screen for therapeutic drugs. Copyright © 2016 by the Genetics Society of America.

  8. The protein kinase TOUSLED is required for maintenance of transcriptional gene silencing in Arabidopsis

    PubMed Central

    Wang, Yu; Liu, Jun; Xia, Ran; Wang, Junguo; Shen, Jie; Cao, Rui; Hong, Xuhui; Zhu, Jian-Kang; Gong, Zhizhong

    2007-01-01

    TOUSLED-like kinases (TLKs) are highly conserved in plants and animals, but direct evidence linking TLKs and transcriptional gene silencing is lacking. We isolated two new alleles of TOUSLED (TSL). Mutations of TSL in ros1 reactivate the transcriptionally silent 35S-NPTII transgene and the transcriptionally silent endogenous loci TSI (TRANSCRIPTIONAL SILENCING INFORMATION). Chromatin immunoprecipitation (ChIP) analysis shows that histone H3Lys9 dimethylation is decreased in the reactivated transgene and endogenous TSI loci in the tsl ros1 mutant. However, there is no change in DNA methylation in the affected loci. Western blot and ChIP assay suggest that TSL might not be responsible for histone H3Ser10 phosphorylation. The tsl seedlings were more sensitive to DNA damage reagent methyl methanesulphonate and UV-B light. Our results provide direct evidence for a crucial role of the TOUSLED protein kinase in the maintenance of transcriptional gene silencing in some genomic regions in a DNA-methylation-independent manner in Arabidopsis. PMID:17110953

  9. The protein kinase TOUSLED is required for maintenance of transcriptional gene silencing in Arabidopsis.

    PubMed

    Wang, Yu; Liu, Jun; Xia, Ran; Wang, Junguo; Shen, Jie; Cao, Rui; Hong, Xuhui; Zhu, Jian-Kang; Gong, Zhizhong

    2007-01-01

    TOUSLED-like kinases (TLKs) are highly conserved in plants and animals, but direct evidence linking TLKs and transcriptional gene silencing is lacking. We isolated two new alleles of TOUSLED (TSL). Mutations of TSL in ros1 reactivate the transcriptionally silent 35S-NPTII transgene and the transcriptionally silent endogenous loci TSI (TRANSCRIPTIONAL SILENCING INFORMATION). Chromatin immunoprecipitation (ChIP) analysis shows that histone H3Lys9 dimethylation is decreased in the reactivated transgene and endogenous TSI loci in the tsl ros1 mutant. However, there is no change in DNA methylation in the affected loci. Western blot and ChIP assay suggest that TSL might not be responsible for histone H3Ser10 phosphorylation. The tsl seedlings were more sensitive to DNA damage reagent methyl methanesulphonate and UV-B light. Our results provide direct evidence for a crucial role of the TOUSLED protein kinase in the maintenance of transcriptional gene silencing in some genomic regions in a DNA-methylation-independent manner in Arabidopsis.

  10. Role of Homer Proteins in the Maintenance of Sleep-Wake States

    PubMed Central

    Naidoo, Nirinjini; Ferber, Megan; Galante, Raymond J.; McShane, Blake; Hu, Jia Hua; Zimmerman, John; Maislin, Greg; Cater, Jacqui; Wyner, Abraham

    2012-01-01

    Sleep is an evolutionarily conserved process that is linked to diurnal cycles and normal daytime wakefulness. Healthy sleep and wakefulness are integral to a healthy lifestyle; this occurs when an organism is able to maintain long bouts of both sleep and wake. Homer proteins, which function as adaptors for group 1 metabotropic glutamate receptors, have been implicated in genetic studies of sleep in both Drosophila and mouse. Drosophila express a single Homer gene product that is upregulated during sleep. By contrast, vertebrates express Homer as both constitutive and immediate early gene (H1a) forms, and H1a is up-regulated during wakefulness. Genetic deletion of Homer in Drosophila results in fragmented sleep and in failure to sustain long bouts of sleep, even under increased sleep drive. However, deletion of Homer1a in mouse results in failure to sustain long bouts of wakefulness. Further evidence for the role of Homer1a in the maintenance of wake comes from the CREB alpha delta mutant mouse, which displays a reduced wake phenotype similar to the Homer1a knockout and fails to up-regulate Homer1a upon sleep loss. Homer1a is a gene whose expression is induced by CREB. Sustained behaviors of the sleep/wake cycle are created by molecular pathways that are distinct from those for arousal or short bouts, and implicate an evolutionarily-conserved role for Homer in sustaining these behaviors. PMID:22532843

  11. Nuclear HMGA1 nonhistone chromatin proteins directly influence mitochondrial transcription, maintenance, and function

    SciTech Connect

    Dement, Gregory A.; Maloney, Scott C.; Reeves, Raymond . E-mail: reevesr@mail.wsu.edu

    2007-01-01

    We have previously demonstrated that HMGA1 proteins translocate from the nucleus to mitochondria and bind to mitochondrial DNA (mtDNA) at the D-loop control region [G.A. Dement, N.R. Treff, N.S. Magnuson, V. Franceschi, R. Reeves, Dynamic mitochondrial localization of nuclear transcription factor HMGA1, Exp. Cell Res. 307 (2005) 388-401.] [11]. To elucidate possible physiological roles for such binding, we employed methods to analyze mtDNA transcription, mitochondrial maintenance, and other organelle functions in transgenic human MCF-7 cells (HA7C) induced to over-express an HA-tagged HMGA1 protein and control (parental) MCF-7 cells. Quantitative real-time (RT) PCR analyses demonstrated that mtDNA levels were reduced approximately 2-fold in HMGA1 over-expressing HA7C cells and flow cytometric analyses further revealed that mitochondrial mass was significantly reduced in these cells. Cellular ATP levels were also reduced in HA7C cells and survival studies showed an increased sensitivity to killing by 2-deoxy-D-glucose, a glycolysis-specific inhibitor. Flow cytometric analyses revealed additional mitochondrial abnormalities in HA7C cells that are consistent with a cancerous phenotype: namely, increased reactive oxygen species (ROS) and increased mitochondrial membrane potential ({delta}{psi}{sub m}). Additional RT-PCR analyses demonstrated that gene transcripts from both the heavy (ND2, COXI, ATP6) and light (ND6) strands of mtDNA were up-regulated approximately 3-fold in HA7C cells. Together, these mitochondrial changes are consistent with many previous reports and reveal several possible mechanisms by which HMGA1 over-expression, a common feature of naturally occurring cancers, may affect tumor progression.

  12. Role of the polarity protein Scribble for podocyte differentiation and maintenance.

    PubMed

    Hartleben, Björn; Widmeier, Eugen; Wanner, Nicola; Schmidts, Miriam; Kim, Sung Tae; Schneider, Lisa; Mayer, Britta; Kerjaschki, Dontscho; Miner, Jeffrey H; Walz, Gerd; Huber, Tobias B

    2012-01-01

    The kidney filter represents a unique assembly of podocyte epithelial cells that tightly enwrap the glomerular capillaries with their complex foot process network. While deficiency of the polarity proteins Crumbs and aPKC result in impaired podocyte foot process architecture, the function of basolateral polarity proteins for podocyte differentiation and maintenance remained unclear. Here we report, that Scribble is expressed in developing podocytes, where it translocates from the lateral aspects of immature podocytes to the basal cell membrane and foot processes of mature podocytes. Immunogold electron microscopy reveals membrane associated localisation of Scribble predominantly at the basolateral site of foot processes. To further study the role of Scribble for podocyte differentiation Scribble(flox/flox) mice were generated by introducing loxP-sites into the Scribble introns 1 and 8 and these mice were crossed to NPHS2.Cre mice and Cre deleter mice. Podocyte-specific Scribble knockout mice develop normally and display no histological, ultrastructural or clinical abnormalities up to 12 months of age. In addition, no increased susceptibility to glomerular stress could be detected in these mice. In contrast, constitutive Scribble knockout animals die during embryonic development indicating the fundamental importance of Scribble for embryogenesis. Like in podocyte-specific Scribble knockout mice, the development of podocyte foot processes and the slit diaphragm was unaffected in kidney cultures from constitutive Scribble knockout animals. In summary these results indicate that basolateral polarity signaling via Scribble is dispensable for podocyte function, highlighting the unique feature of podocyte development with its significant apical membrane expansions being dominated by apical polarity complexes rather than by basolateral polarity signaling.

  13. Minichromosome maintenance 7 protein is a reliable biological marker for human cervical progressive disease

    PubMed Central

    Lobato, Soraya; Tafuri, Alexandre; Fernandes, Paula Ávila; Caliari, Marcelo Vidigal; Silva, Marcos Xavier; Xavier, Marcelo Antônio Pascoal

    2012-01-01

    Objective This study focused on comparing the expression levels of p16, Ki-67, and minichromosome maintenance 7 (MCM7) protein in normal and affected cervical epithelium to ascertain the biological significance of these markers in detecting progressive cervical disease. Methods A quantitative and based on-scanning-microscopy analysis of the three markers expression was performed in normal and cervical intraepithelial neoplasia (CIN) I, II, and III tissues. p16 area as well as p16, Ki-67, and MCM7 positive cells or nuclei were evaluated according to their distribution and extent through the cervical epithelium. Results A clear p16 over-expression was observed in all the dysplastic epithelium tissue samples. The quantitative analysis of p16 area as well as the number of p16 positive cells was able to better discriminate the CIN lesions grades than the usual semi-quantitative analysis. The average Ki-67 labeling indexes for the normal epithelium, CIN I, CIN II, and CIN III groups were 19.8%, 27.3%, 32.8%, and 37.1%, respectively, whereas the mean MCM7 labeling indexes for the correspondent grades were 27.0%, 30.4%, 50.5%, and 67.2%. The Ki-67 and MCM7 labeling indexes were closely correlated with the CIN histological grade, with higher labeling indexe values obtained from the more severe lesions (p<0.05), being the MCM7 labeling indexes the highest values in all the CIN categories (p<0.05). Conclusion We observed a good correlation among the p16, Ki-67, and MCM7 data. In addition, MCM7 demonstrated to be a more efficient and sensitive marker to assess disease progression in the uterine cervix. PMID:22355461

  14. Diagnosis of bladder cancer by immunocytochemical detection of minichromosome maintenance protein-2 in cells retrieved from urine.

    PubMed

    Saeb-Parsy, K; Wilson, A; Scarpini, C; Corcoran, M; Chilcott, S; McKean, M; Thottakam, B; Rai, B; Nabi, G; Rana, D; Perera, M; Stewart, K; Laskey, R A; Neal, D E; Coleman, N

    2012-10-09

    We tested the accuracy of immunocytochemistry (ICC) for minichromosome maintenance protein-2 (MCM-2) in diagnosing bladder cancer, using cells retrieved from urine. Adequate samples were obtained from 497 patients, the majority presenting with gross haematuria (GH) or undergoing cystoscopic surveillance (CS) following previous bladder cancer. We performed an initial study of 313 patients, followed by a validation study of 184 patients. In all cases, presence/absence of bladder cancer was established by cystoscopy/biopsy. In the initial study, receiver operator characteristic analysis showed an area under the curve of 0.820 (P<0.0005) for the GH group and 0.821 (P<0.01) for the CS group. Optimal sensitivity/specificity were provided by threshold values of 50+ MCM-2-positive cells in GH samples and 200+ cells in CS samples, based on a minimum total cell number of 5000. Applying these thresholds to the validation data set gave 81.3% sensitivity, 76.0% specificity and 92.7% negative predictive value (NPV) in GH and 63.2% sensitivity, 89.9% specificity and 89.9% NPV in CS. Minichromosome maintenance protein-2 ICC provided clinically relevant improvements over urine cytology, with greater sensitivity in GH and greater specificity in CS (P=0.05). Minichromosome maintenance protein-2 ICC is a reproducible and accurate test that is suitable for both GH and CS patient groups.

  15. Telomere binding of checkpoint sensor and DNA repair proteins contributes to maintenance of functional fission yeast telomeres.

    PubMed Central

    Nakamura, Toru M; Moser, Bettina A; Russell, Paul

    2002-01-01

    Telomeres, the ends of linear chromosomes, are DNA double-strand ends that do not trigger a cell cycle arrest and yet require checkpoint and DNA repair proteins for maintenance. Genetic and biochemical studies in the fission yeast Schizosaccharomyces pombe were undertaken to understand how checkpoint and DNA repair proteins contribute to telomere maintenance. On the basis of telomere lengths of mutant combinations of various checkpoint-related proteins (Rad1, Rad3, Rad9, Rad17, Rad26, Hus1, Crb2, Chk1, Cds1), Tel1, a telomere-binding protein (Taz1), and DNA repair proteins (Ku70, Rad32), we conclude that Rad3/Rad26 and Tel1/Rad32 represent two pathways required to maintain telomeres and prevent chromosome circularization. Rad1/Rad9/Hus1/Rad17 and Ku70 are two additional epistasis groups, which act in the Rad3/Rad26 pathway. However, Rad3/Rad26 must have additional target(s), as cells lacking Tel1/Rad32, Rad1/Rad9/Hus1/Rad17, and Ku70 groups did not circularize chromosomes. Cells lacking Rad3/Rad26 and Tel1/Rad32 senesced faster than a telomerase trt1Delta mutant, suggesting that these pathways may contribute to telomere protection. Deletion of taz1 did not suppress chromosome circularization in cells lacking Rad3/Rad26 and Tel1/Rad32, also suggesting that two pathways protect telomeres. Chromatin immunoprecipitation analyses found that Rad3, Rad1, Rad9, Hus1, Rad17, Rad32, and Ku70 associate with telomeres. Thus, checkpoint sensor and DNA repair proteins contribute to telomere maintenance and protection through their association with telomeres. PMID:12196391

  16. β-Catenin Accumulation Is Associated With Increased Expression of Nanog Protein and Predicts Maintenance of MSC Self-Renewal.

    PubMed

    Yu, Sang-Jin; Kim, Hyun-Je; Lee, Eui Seok; Park, Chung-Gyu; Cho, Su Jin; Jeon, Soung-Hoo

    2017-02-16

    Human mesenchymal stem cells (hMSCs) are self-renewing cells with the ability to differentiate into organized, functional network of cells. Recent studies have revealed that activation of the Wnt/β-catenin pathway by a glycogen synthase kinase (GSK)-3-specific pharmacological inhibitor, Bio, results in the maintenance of self-renewal in both mouse and human ES cells. The molecular mechanism behind the maintenance of hMSCs by these factors, however, is not fully understood. We found that rEGF enhances the level of β-catenin, a component of the Wnt/β-catenin signaling pathway. Furthermore, it was found that β-catenin upregulates Nanog. EGF activates the β-catenin pathway via the Ras protein and also increased the Nanog protein and gene expression levels 2 h after rEGF treatment. These results suggest that adding EGF can enhance β-catenin and Nanog expression in MSCs and facilitate EGF-mediated maintenance of MSC self-renewal. EGF was shown to augment MSC proliferation while preserving early progenitors within MSC population and thus did not induce differentiation. Thus, EGF not only can be used to expand MSC in vitro but also be utilized to autologous transplantation of MSCs in vivo.

  17. Involvement of protein kinase ζ in the maintenance of hippocampal long-term potentiation in rats with chronic visceral hypersensitivity.

    PubMed

    Chen, Aiqin; Bao, Chengjia; Tang, Ying; Luo, Xiaoqing; Guo, Lixia; Liu, Bin; Lin, Chun

    2015-05-01

    The hippocampal long-term potentiation (LTP) was implicated in the formation of visceral hypersensitivity in rats with irritable bowel syndrome in our previous study. Recent studies have shown that protein kinase M ζ (PKMζ) may be responsible for the maintenance of LTP in memory formation. However, it remains unclear whether PKMζ is involved in the visceral hypersensitivity. In this study, a rat model of visceral hypersensitivity was generated by neonatal maternal separation (NMS). The visceral hypersensitivity was assessed by recording responses of the external oblique abdominal muscle to colorectal distension. Our results demonstrated that hippocampal LTP and visceral hypersensitivity were enhanced significantly in rats of NMS. ζ-Pseudosubstrate inhibitory peptide (ZIP) could dose dependently inhibit the maintenance of Cornu Ammonis area 1 LTP in rats of NMS. Furthermore, Western blot data showed that the expression of hippocampal phosphorylated PKMζ (p-PKMζ) significantly increased in rats of NMS. In addition, bilateral intrahippocampal injections of ZIP attenuated the visceral hypersensitivity dose dependently in rats of NMS. The maximal inhibition was observed at 30 min, and significant inhibition lasted for 1.5-2 h after ZIP application. Besides, data from the open-field test and Morris water maze showed that ZIP did not influence the movement and spatial procedural memory in rats of NMS. In conclusion, p-PKMζ might be a critical protein in the maintenance of hippocampal LTP, which could result in visceral hypersensitivity.

  18. Involvement of protein kinase ζ in the maintenance of hippocampal long-term potentiation in rats with chronic visceral hypersensitivity

    PubMed Central

    Chen, Aiqin; Bao, Chengjia; Tang, Ying; Luo, Xiaoqing; Guo, Lixia; Liu, Bin

    2015-01-01

    The hippocampal long-term potentiation (LTP) was implicated in the formation of visceral hypersensitivity in rats with irritable bowel syndrome in our previous study. Recent studies have shown that protein kinase M ζ (PKMζ) may be responsible for the maintenance of LTP in memory formation. However, it remains unclear whether PKMζ is involved in the visceral hypersensitivity. In this study, a rat model of visceral hypersensitivity was generated by neonatal maternal separation (NMS). The visceral hypersensitivity was assessed by recording responses of the external oblique abdominal muscle to colorectal distension. Our results demonstrated that hippocampal LTP and visceral hypersensitivity were enhanced significantly in rats of NMS. ζ-Pseudosubstrate inhibitory peptide (ZIP) could dose dependently inhibit the maintenance of Cornu Ammonis area 1 LTP in rats of NMS. Furthermore, Western blot data showed that the expression of hippocampal phosphorylated PKMζ (p-PKMζ) significantly increased in rats of NMS. In addition, bilateral intrahippocampal injections of ZIP attenuated the visceral hypersensitivity dose dependently in rats of NMS. The maximal inhibition was observed at 30 min, and significant inhibition lasted for 1.5–2 h after ZIP application. Besides, data from the open-field test and Morris water maze showed that ZIP did not influence the movement and spatial procedural memory in rats of NMS. In conclusion, p-PKMζ might be a critical protein in the maintenance of hippocampal LTP, which could result in visceral hypersensitivity. PMID:25761958

  19. The Intersubunit Bridge B1b of the Bacterial Ribosome Facilitates Initiation of Protein Synthesis and Maintenance of Translational Fidelity.

    PubMed

    Lilleorg, Silva; Reier, Kaspar; Remme, Jaanus; Liiv, Aivar

    2017-04-07

    In bacteria, ribosomal subunits are connected via 12 intersubunit bridges involving RNA-RNA, RNA-protein, and protein-protein interactions. The only protein-protein bridge in the ribosome is ribosomal intersubunit bridge 1b (B1b), which is mainly formed by the bacterial protein L31 (bL31) and connects the head domain of 30S subunit and the central protuberance of the 50S subunit. It is known to be the most dynamic intersubunit bridge. Here, we have evaluated the role of bL31 and thereby the bridge B1b in the working cycle of the ribosome. First, bL31-deficient ribosomes are severely compromised in their ability to ensure translational fidelity particularly in reading frame maintenance in vivo. Second, in the absence of bL31, the rate of initiation is significantly reduced both in vivo and in vitro. Third, polysome profile and subunit reassociation assays demonstrate that bL31 is important for stabilizing subunit joining in vivo and in vitro. Together, our results demonstrate that bL31 is important for determining translational fidelity and stabilizing subunit association. We conclude that the only protein-protein intersubunit bridge of the bacterial ribosome facilitates translation initiation and is essential for maintaining the reading frame of mRNA translation.

  20. Associations between dairy protein intake and body weight and risk markers of diabetes and CVD during weight maintenance.

    PubMed

    Bendtsen, Line Q; Lorenzen, Janne K; Larsen, Thomas M; van Baak, Marleen; Papadaki, Angeliki; Martinez, J Alfredo; Handjieva-Darlenska, Teodora; Jebb, Susan A; Kunešová, Marie; Pfeiffer, Andreas F H; Saris, Wim H M; Astrup, Arne; Raben, Anne

    2014-03-14

    Dairy products have previously been reported to be associated with beneficial effects on body weight and metabolic risk markers. Moreover, primary data from the Diet, Obesity and Genes (DiOGenes) study indicate a weight-maintaining effect of a high-protein-low-glycaemic index diet. The objective of the present study was to examine putative associations between consumption of dairy proteins and changes in body weight and metabolic risk markers after weight loss in obese and overweight adults. Results were based on secondary analyses of data obtained from overweight and obese adults who completed the DiOGenes study. The study consisted of an 8-week weight-loss phase and a 6-month weight-maintenance (WM) phase, where the subjects were given five different diets varying in protein content and glycaemic index. In the present study, data obtained from all the subjects were pooled. Dairy protein intake was estimated from 3 d dietary records at two time points (week 4 and week 26) during the WM phase. Body weight and metabolic risk markers were determined at baseline (week -9 to -11) and before and at the end of the WM phase (week 0 and week 26). Overall, no significant associations were found between consumption of dairy proteins and changes in body weight and metabolic risk markers. However, dairy protein intake tended to be negatively associated with body weight gain (P=0·08; β=-0·17), but this was not persistent when controlled for total protein intake, which indicates that dairy protein adds no additional effect to the effect of total protein. Therefore, the present study does not report that dairy proteins are more favourable than other proteins for body weight regulation.

  1. The role of maintenance proteins in the preservation of epithelial cell identity during mammary gland remodeling and breast cancer initiation.

    PubMed

    Coradini, Danila; Oriana, Saro

    2014-02-01

    During normal postnatal mammary gland development and adult remodeling related to the menstrual cycle, pregnancy, and lactation, ovarian hormones and peptide growth factors contribute to the delineation of a definite epithelial cell identity. This identity is maintained during cell replication in a heritable but DNA-independent manner. The preservation of cell identity is fundamental, especially when cells must undergo changes in response to intrinsic and extrinsic signals. The maintenance proteins, which are required for cell identity preservation, act epigenetically by regulating gene expression through DNA methylation, histone modification, and chromatin remodeling. Among the maintenance proteins, the Trithorax (TrxG) and Polycomb (PcG) group proteins are the best characterized. In this review, we summarize the structures and activities of the TrxG and PcG complexes and describe their pivotal roles in nuclear estrogen receptor activity. In addition, we provide evidence that perturbations in these epigenetic regulators are involved in disrupting epithelial cell identity, mammary gland remodeling, and breast cancer initiation.

  2. Metabolic Turnover of Synaptic Proteins: Kinetics, Interdependencies and Implications for Synaptic Maintenance

    PubMed Central

    Cohen, Laurie D.; Zuchman, Rina; Sorokina, Oksana; Müller, Anke; Dieterich, Daniela C.; Armstrong, J. Douglas; Ziv, Tamar; Ziv, Noam E.

    2013-01-01

    Chemical synapses contain multitudes of proteins, which in common with all proteins, have finite lifetimes and therefore need to be continuously replaced. Given the huge numbers of synaptic connections typical neurons form, the demand to maintain the protein contents of these connections might be expected to place considerable metabolic demands on each neuron. Moreover, synaptic proteostasis might differ according to distance from global protein synthesis sites, the availability of distributed protein synthesis facilities, trafficking rates and synaptic protein dynamics. To date, the turnover kinetics of synaptic proteins have not been studied or analyzed systematically, and thus metabolic demands or the aforementioned relationships remain largely unknown. In the current study we used dynamic Stable Isotope Labeling with Amino acids in Cell culture (SILAC), mass spectrometry (MS), Fluorescent Non–Canonical Amino acid Tagging (FUNCAT), quantitative immunohistochemistry and bioinformatics to systematically measure the metabolic half-lives of hundreds of synaptic proteins, examine how these depend on their pre/postsynaptic affiliation or their association with particular molecular complexes, and assess the metabolic load of synaptic proteostasis. We found that nearly all synaptic proteins identified here exhibited half-lifetimes in the range of 2–5 days. Unexpectedly, metabolic turnover rates were not significantly different for presynaptic and postsynaptic proteins, or for proteins for which mRNAs are consistently found in dendrites. Some functionally or structurally related proteins exhibited very similar turnover rates, indicating that their biogenesis and degradation might be coupled, a possibility further supported by bioinformatics-based analyses. The relatively low turnover rates measured here (∼0.7% of synaptic protein content per hour) are in good agreement with imaging-based studies of synaptic protein trafficking, yet indicate that the metabolic load

  3. Role of G-protein-coupled estrogen receptor (GPER/GPR30) in maintenance of meiotic arrest in fish oocytes.

    PubMed

    Thomas, Peter

    2017-03-01

    An essential role for GPER (formerly known as GPR30) in regulating mammalian reproduction has not been identified to date, although it has shown to be involved in the regulation a broad range of other estrogen-dependent functions. In contrast, an important reproductive role for GPER in the maintenance of oocyte meiotic arrest has been identified in teleost fishes, which is briefly reviewed here. Recent studies have clearly shown that ovarian follicle production of estradiol-17β (E2) maintains meiotic arrest in several teleost species through activation of GPER coupled to a stimulatory G protein (Gs) on oocyte plasma membranes resulting in stimulation of cAMP production and maintenance of elevated cAMP levels. Studies with denuded zebrafish oocytes and with microinjection of GPER antisense oligonucleotides into oocytes have demonstrated the requirement for both ovarian follicle production of estrogens and expression of GPER on the oocyte surface for maintenance of meiotic arrest. This inhibitory action of E2 on the resumption of meiosis is mimicked by the GPER-selective agonist G-1, by the GPER agonists and nuclear ER antagonists, ICI 182,780 and tamoxifen, and also by the xenoestrogen bisphenol-A (BPA) and related alkylphenols. GPER also maintains meiotic arrest of zebrafish oocytes through estrogen- and BPA-dependent GPER activation of epidermal growth factor receptor (EGFR) and mitogen-activated protein kinase (MAPK) signaling. Interestingly, progesterone receptor component 1 (PGRMC1) is also involved in estrogen maintenance of meiotic arrest through regulation of EGFR expression on the oocyte plasma membrane. The preovulatory surge in LH secretion induces the ovarian synthesis of progestin hormones that activate a membrane progestin receptor alpha (mPRα)/inhibitory G protein (Gi) pathway. It also increases ovarian synthesis of the catecholestrogen, 2-hydroxy-estradiol-17β (2-OHE2) which inhibits the GPER/Gs/adenylyl cyclase pathway. Both of these LH actions

  4. Role of protein synthesis and DNA methylation in the consolidation and maintenance of long-term memory in Aplysia.

    PubMed

    Pearce, Kaycey; Cai, Diancai; Roberts, Adam C; Glanzman, David L

    2017-01-09

    Previously, we reported that long-term memory (LTM) in Aplysia can be reinstated by truncated (partial) training following its disruption by reconsolidation blockade and inhibition of PKM (Chen et al., 2014). Here, we report that LTM can be induced by partial training after disruption of original consolidation by protein synthesis inhibition (PSI) begun shortly after training. But when PSI occurs during training, partial training cannot subsequently establish LTM. Furthermore, we find that inhibition of DNA methyltransferase (DNMT), whether during training or shortly afterwards, blocks consolidation of LTM and prevents its subsequent induction by truncated training; moreover, later inhibition of DNMT eliminates consolidated LTM. Thus, the consolidation of LTM depends on two functionally distinct phases of protein synthesis: an early phase that appears to prime LTM; and a later phase whose successful completion is necessary for the normal expression of LTM. Both the consolidation and maintenance of LTM depend on DNA methylation.

  5. Computational comparison of a calcium-dependent jellyfish protein (apoaequorin) and calmodulin-cholesterol in short-term memory maintenance.

    PubMed

    Morrill, Gene A; Kostellow, Adele B; Gupta, Raj K

    2017-03-06

    Memory reconsolidation and maintenance depend on calcium channels and on calcium/calmodulin-dependent kinases regulating protein turnover in the hippocampus. Ingestion of a jellyfish protein, apoaequorin, reportedly protects and/or improves verbal learning in adults and is currently widely advertised for use by the elderly. Apoaequorin is a member of the EF-hand calcium binding family of proteins that includes calmodulin. Calmodulin-1 (148 residues) differs from Apoaequorin (195 residues) in that it contains four rather than three Ca(2+)-binding sites and three rather than four cholesterol-binding (CRAC, CARC) domains. All three cholesterol-binding CARC domains in calmodulin have a high interaction affinity for cholesterol compared to only two high affinity CARC domains in apoaequorin. Both calmodulin and apoaequorin can form dimers with a potential of eight bound Ca(2+) ions and six high affinity-bound cholesterol molecules in calmodulin with six bound Ca(2+) ions and a mixed population of eight cholesterols bound to both CARC and CRAC domains in apoaqueorin. MEMSAT-SVM analysis indicates that both calmodulin and apoaqueorin have a pore-lining region. The Peptide-Cutter algorithm predicts that calmodulin-1 contains 11 trypsin-specific cleavage sites (compared to 21 in apoaqueorin), four of which are potentially blocked by cholesterol and three are within the Ca-binding domains and/or the pore-lining region. Three are clustered between the third and fourth Ca(2+)-binding sites. Only calmodulin pore-lining regions contain Ca(2+) binding sites and as dimers may insert into the plasma membrane of neural cells and act as Ca(2+) channels. In a dietary supplement, bound cholesterol may protect both apoaequorin and calmodulin from proteolysis in the gut as well as facilitate uptake across the blood-brain barrier. Our results suggest that a physiological calmodulin-cholesterol complex, not cholesterol-free jellyfish protein, may better serve as a dietary supplement to

  6. Long term weight maintenance after advice to consume low carbohydrate, higher protein diets--a systematic review and meta analysis.

    PubMed

    Clifton, P M; Condo, D; Keogh, J B

    2014-03-01

    Meta analysis of short term trials indicates that a higher protein, lower carbohydrate weight loss diet enhances fat mass loss and limits lean mass loss compared with a normal protein weight loss diet. Whether this benefit persists long term is not clear. We selected weight loss studies in adults with at least a 12 month follow up in which a higher percentage protein/lower carbohydrate diet was either planned or would be expected for either weight loss or weight maintenance. Studies were selected regardless of the success of the advice but difference in absolute and percentage protein intake at 12 months was used as a moderator in the analysis. Data was analysed using Comprehensive Meta analysis V2 using a random effects analysis. As many as 32 studies with 3492 individuals were analysed with data on fat and lean mass, glucose and insulin from 18 to 22 studies and lipids from 28 studies. A recommendation to consume a lower carbohydrate, higher protein diet in mostly short term intensive interventions with long term follow up was associated with better weight and fat loss but the effect size was small-standardised means of 0.14 and 0.22, p = 0.008 and p < 0.001 respectively (equivalent to 0.4 kg for both). A difference of 5% or greater in percentage protein between diets at 12 mo was associated with a 3 fold greater effect size compared with <5% (p = 0.038) in fat mass (0.9 vs. 0.3 kg). Fasting triglyceride and insulin were also lower with high protein diets with effect sizes of 0.17 and 0.22, p = 0.003 and p = 0.042 respectively. Other lipids and glucose were not different. The short term benefit of higher protein diets appears to persist to a small degree long term. Benefits are greater with better compliance to the diet. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Gap junction proteins: master regulators of the planarian stem cell response to tissue maintenance and injury.

    PubMed

    Peiris, T Harshani; Oviedo, Néstor J

    2013-01-01

    Gap junction (GJ) proteins are crucial mediators of cell-cell communication during embryogenesis, tissue regeneration and disease. GJ proteins form plasma membrane channels that facilitate passage of small molecules across cells and modulate signaling pathways and cellular behavior in different tissues. These properties have been conserved throughout evolution, and in most invertebrates GJ proteins are known as innexins. Despite their critical relevance for physiology and disease, the mechanisms by which GJ proteins modulate cell behavior are poorly understood. This review summarizes findings from recent work that uses planarian flatworms as a paradigm to analyze GJ proteins in the complexity of the whole organism. The planarian model allows access to a large pool of adult somatic stem cells (known as neoblasts) that support physiological cell turnover and tissue regeneration. Innexin proteins are present in planarians and play a fundamental role in controlling neoblast behavior. We discuss the possibility that GJ proteins participate as cellular sensors that inform neoblasts about local and systemic physiological demands. We believe that functional analyses of GJ proteins will bring a complementary perspective to studies that focus on the temporal expression of genes. Finally, integrating functional studies along with molecular genetics and epigenetic approaches would expand our understanding of cellular regulation in vivo and greatly enhance the possibilities for rationally modulating stem cell behavior in their natural environment. This article is part of a Special Issue entitled: The communicating junctions, roles and dysfunctions. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. A structural-maintenance-of-chromosomes hinge domain-containing protein is required for RNA-directed DNA methylation.

    PubMed

    Kanno, Tatsuo; Bucher, Etienne; Daxinger, Lucia; Huettel, Bruno; Böhmdorfer, Gudrun; Gregor, Wolfgang; Kreil, David P; Matzke, Marjori; Matzke, Antonius J M

    2008-05-01

    RNA-directed DNA methylation (RdDM) is a process in which dicer-generated small RNAs guide de novo cytosine methylation at the homologous DNA region. To identify components of the RdDM machinery important for Arabidopsis thaliana development, we targeted an enhancer active in meristems for methylation, which resulted in silencing of a downstream GFP reporter gene. This silencing system also features secondary siRNAs, which trigger methylation that spreads beyond the targeted enhancer region. A screen for mutants defective in meristem silencing and enhancer methylation retrieved six dms complementation groups, which included the known factors DRD1 (ref. 3; a SNF2-like chromatin-remodeling protein) and Pol IVb subunits. Additionally, we identified a previously unknown gene DMS3 (At3g49250), encoding a protein similar to the hinge-domain region of structural maintenance of chromosomes (SMC) proteins. This finding implicates a putative chromosome architectural protein that can potentially link nucleic acids in facilitating an RNAi-mediated epigenetic modification involving secondary siRNAs and spreading of DNA methylation.

  9. Key challenges for the creation and maintenance of specialist protein resources

    PubMed Central

    Holliday, Gemma L; Bairoch, Amos; Bagos, Pantelis G; Chatonnet, Arnaud; Craik, David J; Finn, Robert D; Henrissat, Bernard; Landsman, David; Manning, Gerard; Nagano, Nozomi; O’Donovan, Claire; Pruitt, Kim D; Rawlings, Neil D; Saier, Milton; Sowdhamini, Ramanathan; Spedding, Michael; Srinivasan, Narayanaswamy; Vriend, Gert; Babbitt, Patricia C; Bateman, Alex

    2015-01-01

    As the volume of data relating to proteins increases, researchers rely more and more on the analysis of published data, thus increasing the importance of good access to these data that vary from the supplemental material of individual articles, all the way to major reference databases with professional staff and long-term funding. Specialist protein resources fill an important middle ground, providing interactive web interfaces to their databases for a focused topic or family of proteins, using specialized approaches that are not feasible in the major reference databases. Many are labors of love, run by a single lab with little or no dedicated funding and there are many challenges to building and maintaining them. This perspective arose from a meeting of several specialist protein resources and major reference databases held at the Wellcome Trust Genome Campus (Cambridge, UK) on August 11 and 12, 2014. During this meeting some common key challenges involved in creating and maintaining such resources were discussed, along with various approaches to address them. In laying out these challenges, we aim to inform users about how these issues impact our resources and illustrate ways in which our working together could enhance their accuracy, currency, and overall value. Proteins 2015; 83:1005–1013. © 2015 The Authors. Proteins: Structure, Function, and Bioinformatics Published by Wiley Periodicals, Inc. PMID:25820941

  10. Effect of short-term low-protein diet supplemented with keto acids on hyperphosphatemia in maintenance hemodialysis patients.

    PubMed

    Li, Haiming; Long, Quan; Shao, Chunhai; Fan, Hong; Yuan, Li; Huang, Bihong; Gu, Yong; Lin, Shanyan; Hao, Chuanming; Chen, Jing

    2011-01-01

    To evaluate the effects of short-term restriction of dietary protein intake (DPI) supplemented with keto acids on hyperphosphatemia in maintenance hemodialysis (MHD) patients. Forty MHD patients with uncontrolled hyperphosphatemia were randomized to either low DPI with keto acid-supplemented (sLP) or normal DPI (NP) group for 8 weeks. After 8 weeks, the sLP group was shifted to NP for another 8 weeks. Low-protein diet (LPD) was individualized with total caloric intake 30-35 kcal/kg/day, protein intake of 0.8 g/kg/day and phosphate intake of 500 mg/day. Keto acids were supplied in a dosage of 12 pills per day. Calcium phosphorous metabolism index and nutritional index (serum albumin, total protein, somatometric measurements, 3-day diaries and Mini-Nutritional Assessment score) were recorded. C-reactive protein, CO(2) combining power and Kt/V were measured to evaluate the inflammation, metabolic acidosis and dialysis adequacy, respectively. Serum phosphorus level and calcium-phosphate product were significantly decreased at the end of the first 8 weeks in the sLP group compared to the basal value and the NP group (p < 0.001). No difference was observed in C-reactive protein, Kt/V and nutritional index, while CO(2) combining power was significantly higher at week 8 in the sLP group (p < 0.001). Short-term restriction of DPI supplemented with keto acids could decrease hyperphosphatemia and calcium-phosphate product, while keeping stable nutritional status among MHD patients. Copyright © 2010 S. Karger AG, Basel.

  11. G protein-coupled receptors in stem cell maintenance and somatic reprogramming to pluripotent or cancer stem cells.

    PubMed

    Choi, Hye Yeon; Saha, Subbroto Kumar; Kim, Kyeongseok; Kim, Sangsu; Yang, Gwang-Mo; Kim, BongWoo; Kim, Jin-hoi; Cho, Ssang-Goo

    2015-02-01

    G protein-coupled receptors (GPCRs) are a large class of transmembrane receptors categorized into five distinct families: rhodopsin, secretin, adhesion, glutamate, and frizzled. They bind and regulate 80% of all hormones and account for 20-50% of the pharmaceuticals currently on the market. Hundreds of GPCRs integrate and coordinate the functions of individual cells, mediating signaling between various organs. GPCRs are crucial players in tumor progression, adipogenesis, and inflammation. Several studies have also confirmed their central roles in embryonic development and stem cell maintenance. Recently, GPCRs have emerged as key players in the regulation of cell survival, proliferation, migration, and self-renewal in pluripotent (PSCs) and cancer stem cells (CSCs). Our study and other reports have revealed that the expression of many GPCRs is modulated during the generation of induced PSCs (iPSCs) or CSCs as well as during CSC sphere formation. These GPCRs may have crucial roles in the regulation of selfrenewal and other biological properties of iPSCs and CSCs. This review addresses the current understanding of the role of GPCRs in stem cell maintenance and somatic reprogramming to PSCs or CSCs.

  12. G protein-coupled receptors in stem cell maintenance and somatic reprogramming to pluripotent or cancer stem cells

    PubMed Central

    Choi, Hye Yeon; Saha, Subbroto Kumar; Kim, Kyeongseok; Kim, Sangsu; Yang, Gwang-Mo; Kim, BongWoo; Kim, Jin-hoi; Cho, Ssang-Goo

    2015-01-01

    G protein-coupled receptors (GPCRs) are a large class of transmembrane receptors categorized into five distinct families: rhodopsin, secretin, adhesion, glutamate, and frizzled. They bind and regulate 80% of all hormones and account for 20-50% of the pharmaceuticals currently on the market. Hundreds of GPCRs integrate and coordinate the functions of individual cells, mediating signaling between various organs. GPCRs are crucial players in tumor progression, adipogenesis, and inflammation. Several studies have also confirmed their central roles in embryonic development and stem cell maintenance. Recently, GPCRs have emerged as key players in the regulation of cell survival, proliferation, migration, and self-renewal in pluripotent (PSCs) and cancer stem cells (CSCs). Our study and other reports have revealed that the expression of many GPCRs is modulated during the generation of induced PSCs (iPSCs) or CSCs as well as during CSC sphere formation. These GPCRs may have crucial roles in the regulation of selfrenewal and other biological properties of iPSCs and CSCs. This review addresses the current understanding of the role of GPCRs in stem cell maintenance and somatic reprogramming to PSCs or CSCs. [BMB Reports 2015; 48(2): 68-80] PMID:25413305

  13. Mtr4-like protein coordinates nuclear RNA processing for heterochromatin assembly and for telomere maintenance

    PubMed Central

    Lee, Nathan N.; Chalamcharla, Venkata R.; Reyes-Turcu, Francisca; Mehta, Sameet; Zofall, Martin; Balachandran, Vanivilasini; Dhakshnamoorthy, Jothy; Taneja, Nitika; Yamanaka, Soichiro; Zhou, Ming; Grewal, Shiv I. S.

    2013-01-01

    SUMMARY The regulation of protein-coding and noncoding RNAs is linked to nuclear processes including chromatin modifications and gene silencing. However, the mechanisms that distinguish RNAs and mediate their functions are poorly understood. We describe a nuclear RNA processing network in fission yeast with a core module comprising the Mtr4-like protein, Mtl1, and the zinc finger protein, Red1. The Mtl1-Red1 core promotes degradation of mRNAs and noncoding RNAs, and associates with different proteins to assemble heterochromatin via distinct mechanisms. Mtl1 also forms Red1-independent interactions with evolutionarily conserved proteins named Nrl1 and Ctr1, which associate with splicing factors. Whereas Nrl1 targets transcripts with cryptic introns to form heterochromatin at developmental genes and retrotransposons, Ctr1 functions in processing intron-containing telomerase RNA. Together with our discovery of widespread cryptic introns, including in noncoding RNAs, these findings reveal unique cellular strategies for recognizing regulatory RNAs and coordinating their functions in response to developmental and environmental cues. PMID:24210919

  14. Biochemical characterization of the minichromosome maintenance (MCM) protein of the crenarchaeote Aeropyrum pernix and its interactions with the origin recognition complex (ORC) proteins.

    PubMed

    Atanassova, Neli; Grainge, Ian

    2008-12-16

    Replication in archaea is carried out by proteins that are homologues of eukaryotic counterparts. However, the archaeal systems tend to be much simpler with fewer different genes encoding the core functions than in eukaryotic counterparts. In many archaea, there is a single minichromosome maintenance (MCM) homologue, presumed to be the replicative helicase and between one and three origin recognition complex (ORC) homologues involved in binding to the replication origins. Here we describe the cloning and characterization of the MCM protein from the crenarchaeote Aeropyrum pernix. Like other eukaryotic and archaeal MCM proteins, it is found to be an ATP-dependent DNA helicase, and the putative active site residues involved in ATP binding and hydrolysis are confirmed by mutation. Deletion of the N-terminal 256 amino acids yielded a protein with higher ATPase activity in the absence of DNA and retained robust helicase activity. Interactions with the ORC proteins of A. pernix were examined, and it was found that both ORC homologues could inhibit the helicase activity of MCM. Further it was found that ORC2 could autophosphorylate in the presence of ATP and more remarkably could phosphorylate MCM in a species-specific manner.

  15. Evaluation of Minichromosome Maintenance Protein 7 and c-KIT as Prognostic Markers in Feline Cutaneous Mast Cell Tumours.

    PubMed

    Dobromylskyj, M J; Rasotto, R; Melville, K; Smith, K C; Berlato, D

    2015-11-01

    Mast cell tumours (MCTs) are a common skin tumour in cats, but there is currently no histological grading system or reliable prognostic marker for this species (unlike the situation for dogs). This study utilized a set of 71 feline cutaneous MCTs with known clinical outcomes to assess the potential of various prognostic markers, including the cellular proliferation marker minichromosome maintenance protein (MCM)-7, mitotic index and various KIT labelling characteristics, including KIT positivity, KIT labelling pattern and KIT immunoreactivity score (IS). Of the factors studied, the mitotic index and the KIT labelling pattern were the only features associated significantly with survival times, while the proliferation marker MCM7 and the KIT IS were not. The study also highlights the variability of KIT labelling characteristics between tumours, which may prevent use of this marker as a diagnostic and prognostic tool.

  16. Maintenance energy requirements of beef cows and relationship with cow and calf performance, metabolic hormones, and functional proteins.

    PubMed

    Cooper-Prado, M J; Long, N M; Davis, M P; Wright, E C; Madden, R D; Dilwith, J W; Bailey, C L; Spicer, L J; Wettemann, R P

    2014-08-01

    Gestating Angus, nonlactating, spring-calving cows were used to determine variation in maintenance energy requirements (MR); to evaluate the relationship among MR and cow and calf performance, plasma concentrations of IGF-I, T4, glucose, insulin, and ruminal temperature; and to describe the LM proteome and evaluate protein abundance in cows with different MR. Cows (4 to 7 yr of age) with a BCS of 5.0 ± 0.2 and BW of 582 ± 37 kg in the second to third trimester of gestation were studied in 3 trials (trial 1, n = 23; trial 2, n = 32; trial 3, n = 38). Cows were individually fed a complete diet in amounts to meet predicted MR (Level 1 Model of NRC), and feed intake was adjusted weekly until constant BW was achieved for at least 21 d (maintenance). Cows were classified on the basis of MR as low (>0.5 SD less than mean, LMR), moderate (±0.5 SD of mean, MMR), or high (>0.5 SD more than mean, HMR) MR. Blood samples were taken at maintenance and at 2 mo postpartum in trial 2. Muscle biopsies were taken from LMR and HMR after cows consumed actual MR for 28 d (trial 2) or 21 d (trial 3). Proteins from LM were separated by 2-dimensional difference gel electrophoresis and were identified, and abundance was quantified and compared. The greatest differences in MR between cows were 29%, 24%, and 25% in trials 1, 2, and 3, respectively. Daily MR (NEm, kcal·BW(-0.75)·d(-1)) averaged 89.2 ± 6.3, 93.0 ± 4.9, and 90.4 ± 4.6 in trials 1, 2, and 3, respectively. Postpartum BW and BCS, calf birth and weaning weights, postpartum luteal activity, and ruminal temperature were not influenced by MR of the cows. Concentrations of IGF-I were greater (P = 0.001) in plasma of MMR compared with LMR cows consuming predicted MR diets, and MR was negatively correlated with concentrations of IGF-I in plasma (r = -0.38; P = 0.05) at 2 mo postpartum. A total of 103 proteins were isolated from LM; 52 gene products were identified. Abundance of specific proteins in the LM was not influenced (P > 0

  17. Structure of a double hexamer of the Pyrococcus furiosus minichromosome maintenance protein N-terminal domain

    SciTech Connect

    Meagher, Martin; Enemark, Eric J.

    2016-06-22

    The crystal structure of the N-terminal domain of thePyrococcus furiosusminichromosome maintenance (MCM) protein as a double hexamer is described. The MCM complex is a ring-shaped helicase that unwinds DNA at the replication fork of eukaryotes and archaea. Prior to replication initiation, the MCM complex assembles as an inactive double hexamer at specific sites of DNA. The presented structure is highly consistent with previous MCM double-hexamer structures and shows two MCM hexamers with a head-to-head interaction mediated by the N-terminal domain. Minor differences include a diminished head-to-head interaction and a slightly reduced inter-hexamer rotation.

  18. Cytoplasmic poly (A)-binding protein critically regulates epidermal maintenance and turnover in the planarian Schmidtea mediterranea.

    PubMed

    Bansal, Dhiru; Kulkarni, Jahnavi; Nadahalli, Kavana; Lakshmanan, Vairavan; Krishna, Srikar; Sasidharan, Vidyanand; Geo, Jini; Dilipkumar, Shilpa; Pasricha, Renu; Gulyani, Akash; Raghavan, Srikala; Palakodeti, Dasaradhi

    2017-09-01

    Identifying key cellular events that facilitate stem cell function and tissue organization is crucial for understanding the process of regeneration. Planarians are powerful model system to study regeneration and stem cell (neoblast) function. Here, using planaria, we show that the initial events of regeneration, such as epithelialization and epidermal organization are critically regulated by a novel cytoplasmic poly A-binding protein, SMED-PABPC2. Knockdown of smed-pabpc2 leads to defects in epidermal lineage specification, disorganization of epidermis and ECM, and deregulated wound healing, resulting in the selective failure of neoblast proliferation near the wound region. Polysome profiling suggests that epidermal lineage transcripts, including zfp-1, are translationally regulated by SMED-PABPC2. Together, our results uncover a novel role for SMED-PABPC2 in the maintenance of epidermal and ECM integrity, critical for wound healing and subsequent processes for regeneration. © 2017. Published by The Company of Biologists Ltd.

  19. Expression of chromosomal regional maintenance protein-1 may be associated with subcellular survivin expression in human gastric and colorectal carcinoma

    PubMed Central

    Shintani, Michiko; Tashiro, Akito; Sangawa, Akiko; Yamao, Naoki; Kamoshida, Shingo

    2016-01-01

    Survivin, a member of the inhibitor of apoptosis protein family, is a potential prognostic marker and molecular target for anticancer therapies. Chromosomal regional maintenance protein-1 (CRM-1) mediates the nuclear export of proteins such as survivin. The aims of the present study were to compare the expression and subcellular localization of CRM-1 in human gastric and colorectal carcinomas and to assess the association between CRM-1 and survivin expression in these tumor types. The nuclear and cytoplasmic CRM-1 expression rates in gastric carcinoma were 61% (42/69) and 29% (20/69), respectively, while the nuclear and cytoplasmic CRM-1 expression rates in colorectal carcinoma were 55% (43/78) and 37% (29/78), respectively. Nuclear and cytoplasmic CRM-1 expression was found to be significantly correlated with nuclear and cytoplasmic survivin expression in colorectal carcinoma, but not gastric carcinoma. These results indicate that CRM-1 expression patterns differ between gastric and colorectal carcinomas and thus, we hypothesize that CRM-1-mediated nuclear export of survivin may be deregulated in gastric carcinoma. Therefore, CRM-1 may exhibit different functions in gastric and colorectal carcinoma. PMID:28105170

  20. Evidence for a Role of Arabidopsis CDT1 Proteins in Gametophyte Development and Maintenance of Genome Integrity[C][W

    PubMed Central

    Domenichini, Séverine; Benhamed, Moussa; De Jaeger, Geert; Van De Slijke, Eveline; Blanchet, Sophie; Bourge, Mickaël; De Veylder, Lieven; Bergounioux, Catherine; Raynaud, Cécile

    2012-01-01

    Meristems retain the ability to divide throughout the life cycle of plants, which can last for over 1000 years in some species. Furthermore, the germline is not laid down early during embryogenesis but originates from the meristematic cells relatively late during development. Thus, accurate cell cycle regulation is of utmost importance to avoid the accumulation of mutations during vegetative growth and reproduction. The Arabidopsis thaliana genome encodes two homologs of the replication licensing factor CDC10 Target1 (CDT1), and overexpression of CDT1a stimulates DNA replication. Here, we have investigated the respective functions of Arabidopsis CDT1a and CDT1b. We show that CDT1 proteins have partially redundant functions during gametophyte development and are required for the maintenance of genome integrity. Furthermore, CDT1-RNAi plants show endogenous DNA stress, are more tolerant than the wild type to DNA-damaging agents, and show constitutive induction of genes involved in DNA repair. This DNA stress response may be a direct consequence of reduced CDT1 accumulation on DNA repair or may relate to the ability of CDT1 proteins to form complexes with DNA polymerase ε, which functions in DNA replication and in DNA stress checkpoint activation. Taken together, our results provide evidence for a crucial role of Arabidopsis CDT1 proteins in genome stability. PMID:22773747

  1. Evidence for a role of Arabidopsis CDT1 proteins in gametophyte development and maintenance of genome integrity.

    PubMed

    Domenichini, Séverine; Benhamed, Moussa; De Jaeger, Geert; Van De Slijke, Eveline; Blanchet, Sophie; Bourge, Mickaël; De Veylder, Lieven; Bergounioux, Catherine; Raynaud, Cécile

    2012-07-01

    Meristems retain the ability to divide throughout the life cycle of plants, which can last for over 1000 years in some species. Furthermore, the germline is not laid down early during embryogenesis but originates from the meristematic cells relatively late during development. Thus, accurate cell cycle regulation is of utmost importance to avoid the accumulation of mutations during vegetative growth and reproduction. The Arabidopsis thaliana genome encodes two homologs of the replication licensing factor CDC10 Target1 (CDT1), and overexpression of CDT1a stimulates DNA replication. Here, we have investigated the respective functions of Arabidopsis CDT1a and CDT1b. We show that CDT1 proteins have partially redundant functions during gametophyte development and are required for the maintenance of genome integrity. Furthermore, CDT1-RNAi plants show endogenous DNA stress, are more tolerant than the wild type to DNA-damaging agents, and show constitutive induction of genes involved in DNA repair. This DNA stress response may be a direct consequence of reduced CDT1 accumulation on DNA repair or may relate to the ability of CDT1 proteins to form complexes with DNA polymerase ε, which functions in DNA replication and in DNA stress checkpoint activation. Taken together, our results provide evidence for a crucial role of Arabidopsis CDT1 proteins in genome stability.

  2. Protein Kinase D Controls the Integrity of Golgi Apparatus and the Maintenance of Dendritic Arborization in Hippocampal Neurons

    PubMed Central

    Czöndör, Katalin; Ellwanger, Kornelia; Fuchs, Yannick F.; Lutz, Sylke; Gulyás, Márton; Mansuy, Isabelle M.; Hausser, Angelika; Pfizenmaier, Klaus

    2009-01-01

    Protein kinase D (PKD) is known to participate in various cellular functions, including secretory vesicle fission from the Golgi and plasma membrane-directed transport. Here, we report on expression and function of PKD in hippocampal neurons. Expression of an enhanced green fluorescent protein (EGFP)-tagged PKD activity reporter in mouse embryonal hippocampal neurons revealed high endogenous PKD activity at the Golgi complex and in the dendrites, whereas PKD activity was excluded from the axon in parallel with axonal maturation. Expression of fluorescently tagged wild-type PKD1 and constitutively active PKD1S738/742E (caPKD1) in neurons revealed that both proteins were slightly enriched at the trans-Golgi network (TGN) and did not interfere with its thread-like morphology. By contrast, expression of dominant-negative kinase inactive PKD1K612W (kdPKD1) led to the disruption of the neuronal Golgi complex, with kdPKD1 strongly localized to the TGN fragments. Similar findings were obtained from transgenic mice with inducible, neuron-specific expression of kdPKD1-EGFP. As a prominent consequence of kdPKD1 expression, the dendritic tree of transfected neurons was reduced, whereas caPKD1 increased dendritic arborization. Our results thus provide direct evidence that PKD activity is selectively involved in the maintenance of dendritic arborization and Golgi structure of hippocampal neurons. PMID:19211839

  3. Relatively high-protein or 'low-carb' energy-restricted diets for body weight loss and body weight maintenance?

    PubMed

    Soenen, Stijn; Bonomi, Alberto G; Lemmens, Sofie G T; Scholte, Jolande; Thijssen, Myriam A M A; van Berkum, Frank; Westerterp-Plantenga, Margriet S

    2012-10-10

    'Low-carb' diets have been suggested to be effective in body weight (BW) management. However, these diets are relatively high in protein as well. To unravel whether body-weight loss and weight-maintenance depends on the high-protein or the 'low-carb' component of the diet. Body-weight (BW), fat mass (FM), blood- and urine-parameters of 132 participants (age=50 ± 12 yr; BW=107 ± 20 kg; BMI=37 ± 6 kg/m(2); FM=47.5 ± 11.9 kg) were compared after 3 and 12 months between four energy-restricted diets with 33% of energy requirement for the first 3 months, and 67% for the last 9 months: normal-protein normal-carbohydrate (NPNC), normal-protein low-carbohydrate (NPLC); high-protein normal-carbohydrate (HPNC), high-protein low-carbohydrate (HPLC); 24h N-analyses confirmed daily protein intakes for the normal-protein diets of 0.7 ± 0.1 and for the high-protein diets of 1.1 ± 0.2g/kg BW (p<0.01). BW and FM decreased over 3 months (p<0.001): HP (-14.1 ± 4 kg; -11.9 ± 1.7 kg) vs. NP (-11.5 ± 4 kg; -9.3 ± 0.7 kg) (p<0.001); LC (-13.5 ± 4 kg; -11.0 ± 1.2 kg) vs. NC (-12.3 ± 3 kg; -10.3 ± 1.1 kg) (ns). Diet × time interaction showed HPLC (-14.7 ± 5 kg; -11.9 ± 1.6 kg) vs. HPNC (-13.8 ± 3 kg; -11.9 ± 1.8 kg) (ns); NPLC (-12.2 ± 4 kg; -10.0 ± 0.8 kg) vs. NPNC (-10.7 ± 4 kg; -8.6 ± 0.7 kg) (ns); HPLC vs. NPLC (p<0.001); HPNC vs. NPNC (p<0.001). Decreases over 12 months (p<0.001) showed HP (-12.8 ± 4 kg; -9.1 ± 0.8 kg) vs. NP (-8.9 ± 3 kg; -7.7 ± 0.6 kg) (p<0.001); LC (-10.6 ± 4 kg; -8.3 ± 0.7 kg) vs. NC (11.1 ± 3 kg; 9.3 ± 0.7 kg) (ns). Diet × time interaction showed HPLC (-11.6 ± 5 kg ; -8.2 ± 0.7 kg) vs. HPNC (-14.1 ± 4 kg; -10.0 ± 0.9 kg) (ns); NPNC (-8.2 ± 3 kg; -6.7 ± 0.6 kg) vs. NPLC (-9.7 ± 3 kg; -8.5 ± 0.7 kg) (ns); HPLC vs. NPLC (p<0.01); HPNC vs. NPNC (p<0.01). HPNC vs. all other diets reduced diastolic blood pressure more. Relationships between changes in BW, FM, FFM or metabolic parameters and energy percentage of fat in the diet

  4. Maintenance of a Protein Structure in the Dynamic Evolution of TIMPs over 600 Million Years

    PubMed Central

    Nicosia, Aldo; Maggio, Teresa; Costa, Salvatore; Salamone, Monica; Tagliavia, Marcello; Mazzola, Salvatore; Gianguzza, Fabrizio; Cuttitta, Angela

    2016-01-01

    Deciphering the events leading to protein evolution represents a challenge, especially for protein families showing complex evolutionary history. Among them, TIMPs represent an ancient eukaryotic protein family widely distributed in the animal kingdom. They are known to control the turnover of the extracellular matrix and are considered to arise early during metazoan evolution, arguably tuning essential features of tissue and epithelial organization. To probe the structure and molecular evolution of TIMPs within metazoans, we report the mining and structural characterization of a large data set of TIMPs over approximately 600 Myr. The TIMPs repertoire was explored starting from the Cnidaria phylum, coeval with the origins of connective tissue, to great apes and humans. Despite dramatic sequence differences compared with highest metazoans, the ancestral proteins displayed the canonical TIMP fold. Only small structural changes, represented by an α-helix located in the N-domain, have occurred over the evolution. Both the occurrence of such secondary structure elements and the relative solvent accessibility of the corresponding residues in the three-dimensional structures raises the possibility that these sites represent unconserved element prone to accept variations. PMID:26957029

  5. Levels of the E2 interacting protein TopBP1 modulate papillomavirus maintenance stage replication

    SciTech Connect

    Kanginakudru, Sriramana; DeSmet, Marsha; Thomas, Yanique; Morgan, Iain M.; Androphy, Elliot J.

    2015-04-15

    The evolutionarily conserved DNA topoisomerase II beta-binding protein 1 (TopBP1) functions in DNA replication, DNA damage response, and cell survival. We analyzed the role of TopBP1 in human and bovine papillomavirus genome replication. Consistent with prior reports, TopBP1 co-localized in discrete nuclear foci and was in complex with papillomavirus E2 protein. Similar to E2, TopBP1 is recruited to the region of the viral origin of replication during G1/S and early S phase. TopBP1 knockdown increased, while over-expression decreased transient virus replication, without affecting cell cycle. Similarly, using cell lines harboring HPV-16 or HPV-31 genome, TopBP1 knockdown increased while over-expression reduced viral copy number relative to genomic DNA. We propose a model in which TopBP1 serves dual roles in viral replication: it is essential for initiation of replication yet it restricts viral copy number. - Highlights: • Protein interaction study confirmed In-situ interaction between TopBP1 and E2. • TopBP1 present at papillomavirus ori in G1/S and early S phase of cell cycle. • TopBP1 knockdown increased, over-expression reduced virus replication. • TopBP1 protein level change did not influence cell survival or cell cycle. • TopBP1 displaced from papillomavirus ori after initiation of replication.

  6. Concise review: Fragile X proteins in stem cell maintenance and differentiation.

    PubMed

    Li, Yue; Zhao, Xinyu

    2014-07-01

    Fragile X syndrome (FXS), the most common genetic form of autism spectrum disorder, is caused by deficiency of the fragile X mental retardation protein (FMRP). Despite extensive research and scientific progress, understanding how FMRP regulates brain development and function remains a major challenge. FMRP is a neuronal RNA-binding protein that binds about a third of messenger RNAs in the brain and controls their translation, stability, and cellular localization. The absence of FMRP results in increased protein synthesis, leading to enhanced signaling in a number of intracellular pathways, including the mTOR, mGLuR5, ERK, Gsk3β, PI3K, and insulin pathways. Until recently, FXS was largely considered a deficit of mature neurons; however, a number of new studies have shown that FMRP may also play important roles in stem cells, among them neural stem cells, germline stem cells, and pluripotent stem cells. In this review, we will cover these newly discovered functions of FMRP, as well as the other two fragile X-related proteins, in stem cells. We will also discuss the literature on the use of stem cells, particularly neural stem cells and induced pluripotent stem cells, as model systems for studying the functions of FMRP in neuronal development.

  7. Key challenges for the creation and maintenance of specialist protein resources.

    PubMed

    Holliday, Gemma L; Bairoch, Amos; Bagos, Pantelis G; Chatonnet, Arnaud; Craik, David J; Finn, Robert D; Henrissat, Bernard; Landsman, David; Manning, Gerard; Nagano, Nozomi; O'Donovan, Claire; Pruitt, Kim D; Rawlings, Neil D; Saier, Milton; Sowdhamini, Ramanathan; Spedding, Michael; Srinivasan, Narayanaswamy; Vriend, Gert; Babbitt, Patricia C; Bateman, Alex

    2015-06-01

    As the volume of data relating to proteins increases, researchers rely more and more on the analysis of published data, thus increasing the importance of good access to these data that vary from the supplemental material of individual articles, all the way to major reference databases with professional staff and long-term funding. Specialist protein resources fill an important middle ground, providing interactive web interfaces to their databases for a focused topic or family of proteins, using specialized approaches that are not feasible in the major reference databases. Many are labors of love, run by a single lab with little or no dedicated funding and there are many challenges to building and maintaining them. This perspective arose from a meeting of several specialist protein resources and major reference databases held at the Wellcome Trust Genome Campus (Cambridge, UK) on August 11 and 12, 2014. During this meeting some common key challenges involved in creating and maintaining such resources were discussed, along with various approaches to address them. In laying out these challenges, we aim to inform users about how these issues impact our resources and illustrate ways in which our working together could enhance their accuracy, currency, and overall value.

  8. Different phenotypes in vivo are associated with ATPase motif mutations in Schizosaccharomyces pombe minichromosome maintenance proteins.

    PubMed Central

    Gómez, Eliana B; Catlett, Michael G; Forsburg, Susan L

    2002-01-01

    The six conserved MCM proteins are essential for normal DNA replication. They share a central core of homology that contains sequences related to DNA-dependent and AAA(+) ATPases. It has been suggested that the MCMs form a replicative helicase because a hexameric subcomplex formed by MCM4, -6, and -7 proteins has in vitro DNA helicase activity. To test whether ATPase and helicase activities are required for MCM protein function in vivo, we mutated conserved residues in the Walker A and Walker B motifs of MCM4, -6, and -7 and determined that equivalent mutations in these three proteins have different in vivo effects in fission yeast. Some mutations reported to abolish the in vitro helicase activity of the mouse MCM4/6/7 subcomplex do not affect the in vivo function of fission yeast MCM complex. Mutations of consensus CDK sites in Mcm4p and Mcm7p also have no phenotypic consequences. Co-immunoprecipitation analyses and in situ chromatin-binding experiments were used to study the ability of the mutant Mcm4ps to associate with the other MCMs, localize to the nucleus, and bind to chromatin. We conclude that the role of ATP binding and hydrolysis is different for different MCM subunits. PMID:11973289

  9. Determinants of the serum concentrations of low molecular weight proteins in patients on maintenance hemodialysis.

    PubMed

    Kabanda, A; Jadoul, M; Pochet, J M; Lauwerys, R; van Ypersele de Strihou, C; Bernard, A

    1994-06-01

    Factors influencing the serum concentrations of low molecular weight proteins (LMWP) during long-term hemodialysis were studied in 112 patients undergoing dialysis for an average of 61.1 months (range 1 to 243). These patients were treated with AN69, cellulose acetate, cuprophan or polysulfone membranes. The following proteins were measured in serum before and after a four hour dialysis session: cystatin C (CYST C), beta 2-microglobulin (beta 2 m), Clara cell protein (CC16) and retinol-binding protein (RBP). Predialysis levels of the four proteins were markedly elevated. In simple regression analysis, pre-dialysis serum concentrations of beta 2 m and CC16 weakly correlated with the duration of dialysis treatment, but these relations completely disappeared when a stepwise regression analysis was performed using as predictors age, sex, residual diuresis, body weight loss (BWL), duration of hemodialysis and the type or ultrafiltration coefficient (UFC) of the membranes. The only significant determinants which emerged from this analysis were the residual diuresis and age which negatively correlated with CYST C, beta 2m and CC16 (residual diuresis only), and sex which influenced CYST C. During the dialysis session, the microproteins underwent changes that were related to their molecular radius, the membrane UFC and the BWL. After adjustment for the latter, high flux membranes (UFC > or = 15 ml/h.m2.mm Hg) allowed up to 50% of CYST C and 25% of beta 2m to be removed. No significant elimination of CC16 and RBP was evident. On the basis of these results, we estimated the effective pore radius of high flux membranes between 1.5 and 1.7 nm and that of low flux membranes as below 1.5 nm.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. CEP152 is a genome maintenance protein disrupted in Seckel syndrome.

    PubMed

    Kalay, Ersan; Yigit, Gökhan; Aslan, Yakup; Brown, Karen E; Pohl, Esther; Bicknell, Louise S; Kayserili, Hülya; Li, Yun; Tüysüz, Beyhan; Nürnberg, Gudrun; Kiess, Wieland; Koegl, Manfred; Baessmann, Ingelore; Buruk, Kurtulus; Toraman, Bayram; Kayipmaz, Saadettin; Kul, Sibel; Ikbal, Mevlit; Turner, Daniel J; Taylor, Martin S; Aerts, Jan; Scott, Carol; Milstein, Karen; Dollfus, Helene; Wieczorek, Dagmar; Brunner, Han G; Hurles, Matthew; Jackson, Andrew P; Rauch, Anita; Nürnberg, Peter; Karagüzel, Ahmet; Wollnik, Bernd

    2011-01-01

    Functional impairment of DNA damage response pathways leads to increased genomic instability. Here we describe the centrosomal protein CEP152 as a new regulator of genomic integrity and cellular response to DNA damage. Using homozygosity mapping and exome sequencing, we identified CEP152 mutations in Seckel syndrome and showed that impaired CEP152 function leads to accumulation of genomic defects resulting from replicative stress through enhanced activation of ATM signaling and increased H2AX phosphorylation.

  11. CEP152 is a genome maintenance protein disrupted in Seckel syndrome

    PubMed Central

    Kalay, Ersan; Yigit, Gökhan; Aslan, Yakup; Brown, Karen E; Pohl, Esther; Bicknell, Louise S; Kayserili, Hülya; Li, Yun; Tüysüz, Beyhan; Nürnberg, Gudrun; Kiess, Wieland; Koegl, Manfred; Baessmann, Ingelore; Buruk, Kurtulus; Toraman, Bayram; Kayipmaz, Saadettin; Kul, Sibel; Ikbal, Mevlit; Turner, Daniel J; Taylor, Martin S; Aerts, Jan; Scott, Carol; Milstein, Karen; Dollfus, Helene; Wieczorek, Dagmar; Brunner, Han G; Hurles, Matthew; Jackson, Andrew P; Rauch, Anita; Nürnberg, Peter; Karagüzel, Ahmet; Wollnik, Bernd

    2012-01-01

    Functional impairment of DNA damage response pathways leads to increased genomic instability. Here we describe the centrosomal protein CEP152 as a new regulator of genomic integrity and cellular response to DNA damage. Using homozygosity mapping and exome sequencing, we identified CEP152 mutations in Seckel syndrome and showed that impaired CEP152 function leads to accumulation of genomic defects resulting from replicative stress through enhanced activation of ATM signaling and increased H2AX phosphorylation. PMID:21131973

  12. In vivo requirement of protein prenylation for maintenance of retinal cytoarchitecture and photoreceptor structure

    PubMed Central

    1995-01-01

    Recent studies have demonstrated that inhibition of mevalonate synthesis in cultured cells leads to altered cell morphology due to inhibition of protein prenylation. To investigate the effects in vivo of mevalonate deprivation in nondividing, terminally differentiated neural cells, we have analyzed the effects on retinal tissue of intravitreal injection of lovastatin, a potent inhibitor of the mevalonate-producing enzyme, HMG-CoA reductase. A single injection of lovastatin (0.25 mumol) produced profound dysplastic-like changes in adult rat retinas primarily involving the photoreceptor layer. Within 2 d after injection, photoreceptor nuclei migrated in a circular pattern resulting in the formation of rosette-like structures by 4 d. Also during this period, photoreceptor inner and outer segment degeneration was evident. By 21 d, intact photoreceptor nuclei with remnants of inner and outer segments were dispersed throughout all retinal layers. To investigate the biochemical specificity of the lovastatin-induced alterations, and to distinguish the relative importance of the various branches of the mevalonate pathway, the incorporation of [3H]acetate into retinal lipids was examined in the presence and absence of metabolic inhibitors. HPLC analysis of lovastatin-treated retinas revealed a dramatic reduction in the incorporation of intravitreally injected [3H]acetate into nonsaponifiable lipids, compared with controls. In contrast, intravitreal injection of NB-598, a specific inhibitor of squalene epoxidase, eliminated the conversion of newly synthesized squalene to sterols without obvious pathology. Hence, involvement to the sterol branch of isoprenoid metabolism in the lovastatin-induced morphologic disruption was obviated. Intravitreal injection of 0.27 mumol of N-acetyl-S-trans,trans-farnesyl-L-cysteine (AFC), an inhibitor of carboxyl methyltransferase activity and prenylated protein function, produced morphologic changes that were virtually indistinguishable from

  13. Maintaining protein homeostasis: early and late endosomal dual recycling for the maintenance of intracellular pools of the plasma membrane protein Chs3

    PubMed Central

    Arcones, Irene; Sacristán, Carlos; Roncero, Cesar

    2016-01-01

    The major chitin synthase activity in yeast cells, Chs3, has become a paradigm in the study of the intracellular traffic of transmembrane proteins due to its tightly regulated trafficking. This includes an efficient mechanism for the maintenance of an extensive reservoir of Chs3 at the trans-Golgi network/EE, which allows for the timely delivery of the protein to the plasma membrane. Here we show that this intracellular reservoir of Chs3 is maintained not only by its efficient AP-1–mediated recycling, but also by recycling through the retromer complex, which interacts with Chs3 at a defined region in its N-terminal cytosolic domain. Moreover, the N-terminal ubiquitination of Chs3 at the plasma membrane by Rsp5/Art4 distinctly labels the protein and regulates its retromer-mediated recycling by enabling Chs3 to be recognized by the ESCRT machinery and degraded in the vacuole. Therefore the combined action of two independent but redundant endocytic recycling mechanisms, together with distinct labels for vacuolar degradation, determines the final fate of the intracellular traffic of the Chs3 protein, allowing yeast cells to regulate morphogenesis, depending on environmental constraints. PMID:27798229

  14. Nutritional Status of Maintenance Dialysis Patients: Low Lean Body Mass Index and Obesity Are Common, Protein-Energy Wasting Is Uncommon.

    PubMed

    Koefoed, Mette; Kromann, Charles Boy; Juliussen, Sophie Ryberg; Hvidtfeldt, Danni; Ekelund, Bo; Frandsen, Niels Erik; Marckmann, Peter

    2016-01-01

    Maintenance dialysis patients are at increased risk of abnormal nutritional status due to numerous causative factors, both nutritional and non-nutritional. The present study assessed the current prevalence of protein-energy wasting, low lean body mass index and obesity in maintenance dialysis patients, and compared different methods of nutritional assessment. In a cross-sectional study conducted in 2014 at Roskilde Hospital, Denmark, we performed anthropometry (body weight, skinfolds, mid-arm, waist, and hip circumferences), and determined plasma albumin and normalized protein catabolic rate in order to assess the prevalence of protein-energy wasting, low lean body mass index and obesity in these patients. Seventy-nine eligible maintenance dialysis patients participated. The prevalence of protein-energy wasted patients was 4% (95% CI: 2-12) as assessed by the coexistence of low lean body mass index and low fat mass index. Low lean body mass index was seen in 32% (95% CI: 22-44). Obesity prevalence as assessed from fat mass index was 43% (95% CI: 32-55). Coexistence of low lean body mass index and obesity was seen in 10% (95% CI: 5-19). The prevalence of protein-energy wasting and obesity varied considerably, depending on nutritional assessment methodology. Our data indicate that protein-energy wasting is uncommon, whereas low lean body mass index and obesity are frequent conditions among patients in maintenance dialysis. A focus on how to increase and preserve lean body mass in dialysis patients is suggested in the future. In order to clearly distinguish between shortage, sufficiency and abundance of protein and/or fat deposits in maintenance dialysis patients, we suggest the simple measurements of lean body mass index and fat mass index.

  15. Nutritional Status of Maintenance Dialysis Patients: Low Lean Body Mass Index and Obesity Are Common, Protein-Energy Wasting Is Uncommon

    PubMed Central

    Koefoed, Mette; Kromann, Charles Boy; Juliussen, Sophie Ryberg; Hvidtfeldt, Danni; Ekelund, Bo; Frandsen, Niels Erik; Marckmann, Peter

    2016-01-01

    Background and Aims Maintenance dialysis patients are at increased risk of abnormal nutritional status due to numerous causative factors, both nutritional and non-nutritional. The present study assessed the current prevalence of protein-energy wasting, low lean body mass index and obesity in maintenance dialysis patients, and compared different methods of nutritional assessment. Methods In a cross-sectional study conducted in 2014 at Roskilde Hospital, Denmark, we performed anthropometry (body weight, skinfolds, mid-arm, waist, and hip circumferences), and determined plasma albumin and normalized protein catabolic rate in order to assess the prevalence of protein-energy wasting, low lean body mass index and obesity in these patients. Results Seventy-nine eligible maintenance dialysis patients participated. The prevalence of protein-energy wasted patients was 4% (95% CI: 2–12) as assessed by the coexistence of low lean body mass index and low fat mass index. Low lean body mass index was seen in 32% (95% CI: 22–44). Obesity prevalence as assessed from fat mass index was 43% (95% CI: 32–55). Coexistence of low lean body mass index and obesity was seen in 10% (95% CI: 5–19). The prevalence of protein-energy wasting and obesity varied considerably, depending on nutritional assessment methodology. Conclusions Our data indicate that protein-energy wasting is uncommon, whereas low lean body mass index and obesity are frequent conditions among patients in maintenance dialysis. A focus on how to increase and preserve lean body mass in dialysis patients is suggested in the future. In order to clearly distinguish between shortage, sufficiency and abundance of protein and/or fat deposits in maintenance dialysis patients, we suggest the simple measurements of lean body mass index and fat mass index. PMID:26919440

  16. Estimation of protein requirement for maintenance in adult parrots (Amazona spp.) by determining inevitable N losses in excreta.

    PubMed

    Westfahl, C; Wolf, P; Kamphues, J

    2008-06-01

    Especially in older pet birds, an unnecessary overconsumption of protein--presumably occurring in human custody--should be avoided in view of a potential decrease in the excretory organs' (liver, kidney) efficiency. Inevitable nitrogen (N)-losses enable the estimation of protein requirement for maintenance, because these losses have at least to be replaced to maintain N equilibrium. To determine the inevitable N losses in excreta of adult amazons (Amazona spp.), a frugivor-granivorous avian species from South America, adult amazons (n = 8) were fed a synthetic nearly N-free diet (in dry matter; DM: 37.8% starch, 26.6% sugar, 11.0% fat) for 9 days. Throughout the trial, feed and water intake were recorded, the amounts of excreta were measured and analysed for DM and ash content, N (Dumas analysis) and uric acid (enzymatic-photometric analysis) content. Effects of the N-free diet on body weight (BW) and protein-related blood parameters were quantified and compared with data collected during a previous 4-day period in which a commercial seed mixture was offered to the birds. After feeding an almost N-free diet for 9 days, under the conditions of a DM intake (20.1 g DM/bird/day) as in seeds and digestibility of organic matter comparable with those when fed seeds (82% and 76% respectively), it was possible to quantify the inevitable N losses via excrements to be 87.2 mg/bird/day or 172.5 mg/kg BW(0.75)/day. Assuming a utilization coefficient of 0.57 this leads to an estimated protein need of approximately 1.9 g/kg BW(0.75)/day (this value does not consider further N losses via feathers and desquamated cells; with the prerequisite that there is a balanced amino acid pattern).

  17. Both Maintenance and Avoidance of RNA-Binding Protein Interactions Constrain Coding Sequence Evolution.

    PubMed

    Savisaar, Rosina; Hurst, Laurence D

    2017-05-01

    While the principal force directing coding sequence (CDS) evolution is selection on protein function, to ensure correct gene expression CDSs must also maintain interactions with RNA-binding proteins (RBPs). Understanding how our genes are shaped by these RNA-level pressures is necessary for diagnostics and for improving transgenes. However, the evolutionary impact of the need to maintain RBP interactions remains unresolved. Are coding sequences constrained by the need to specify RBP binding motifs? If so, what proportion of mutations are affected? Might sequence evolution also be constrained by the need not to specify motifs that might attract unwanted binding, for instance because it would interfere with exon definition? Here, we have scanned human CDSs for motifs that have been experimentally determined to be recognized by RBPs. We observe two sets of motifs-those that are enriched over nucleotide-controlled null and those that are depleted. Importantly, the depleted set is enriched for motifs recognized by non-CDS binding RBPs. Supporting the functional relevance of our observations, we find that motifs that are more enriched are also slower-evolving. The net effect of this selection to preserve is a reduction in the over-all rate of synonymous evolution of 2-3% in both primates and rodents. Stronger motif depletion, on the other hand, is associated with stronger selection against motif gain in evolution. The challenge faced by our CDSs is therefore not only one of attracting the right RBPs but also of avoiding the wrong ones, all while also evolving under selection pressures related to protein structure. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  18. Membrane Frizzled Related Protein is necessary for the normal development and maintenance of photoreceptor outer segments

    PubMed Central

    Won, Jungyeon; Smith, Richard S.; Peachey, Neal S.; Wu, Jiang; Hicks, Wanda L.; Naggert, Jürgen K.; Nishina, Patsy M.

    2009-01-01

    A 4 base pair deletion in a splice donor site of the Mfrp (membrane-type frizzled-related protein) gene, herein referred to as Mfrprd6/rd6, is predicted to lead to the skipping of exon 4 and photoreceptor degeneration in retinal degeneration 6 (rd6) mutant mice. Little, however, is known about the function of the protein or how the mutation causes the degenerative retinal phenotype. Here we examine ultrastructural changes in the retina of Mfrprd6/rd6 mice to determine the earliest effects of the mutation. We also extend the reported observations of the expression pattern of the dicistronic Mfrp/C1qtnf5 message and the localization of these and other retinal pigment epithelium (RPE) and retinal proteins during development and assess the ability of RPE cells to phagocytize outer segments in mutant and WT mice. At the ultrastructural level, outer segments do not develop normally in Mfrprd6/rd6 mutants. They are disorganized and become progressively shorter as mutant mice age. Additionally, there are focal areas in which there is a reduction of apical RPE microvilli. At P25, the rod ERG a-wave of Mfrprd6/rd6 mice is reduced in amplitude by ~50% as are ERG components generated by the RPE. Examination of β-catenin localization and Fos and Tcf-1 expression, intermediates of the canonical Wnt-pathway, showed they were not different between mutant and WT mice, suggesting that MFRP may operate through an alternative pathway. Finally, impaired outer segment phagocytosis was observed in Mfrprd6/rd6 mice both in standard ambient lighting conditions and with bright light exposure when compared to WT controls. PMID:18764959

  19. Role of protein synthesis and DNA methylation in the consolidation and maintenance of long-term memory in Aplysia

    PubMed Central

    Pearce, Kaycey; Cai, Diancai; Roberts, Adam C; Glanzman, David L

    2017-01-01

    Previously, we reported that long-term memory (LTM) in Aplysia can be reinstated by truncated (partial) training following its disruption by reconsolidation blockade and inhibition of PKM (Chen et al., 2014). Here, we report that LTM can be induced by partial training after disruption of original consolidation by protein synthesis inhibition (PSI) begun shortly after training. But when PSI occurs during training, partial training cannot subsequently establish LTM. Furthermore, we find that inhibition of DNA methyltransferase (DNMT), whether during training or shortly afterwards, blocks consolidation of LTM and prevents its subsequent induction by truncated training; moreover, later inhibition of DNMT eliminates consolidated LTM. Thus, the consolidation of LTM depends on two functionally distinct phases of protein synthesis: an early phase that appears to prime LTM; and a later phase whose successful completion is necessary for the normal expression of LTM. Both the consolidation and maintenance of LTM depend on DNA methylation. DOI: http://dx.doi.org/10.7554/eLife.18299.001 PMID:28067617

  20. Kruppel-like zinc finger protein Glis2 is essential for the maintenance of normal renal functions.

    PubMed

    Kim, Yong-Sik; Kang, Hong Soon; Herbert, Ronald; Beak, Ju Youn; Collins, Jennifer B; Grissom, Sherry F; Jetten, Anton M

    2008-04-01

    To obtain insight into the physiological functions of the Krüppel-like zinc finger protein Gli-similar 2 (Glis2), mice deficient in Glis2 expression were generated. Glis2 mutant (Glis2(mut)) mice exhibit significantly shorter life spans than do littermate wild-type (WT) mice due to the development of progressive chronic kidney disease with features resembling nephronophthisis. Glis2(mut) mice develop severe renal atrophy involving increased cell death and basement membrane thickening in the proximal convoluted tubules. This development is accompanied by infiltration of lymphocytic inflammatory cells and interstitial/glomerular fibrosis. The severity of the fibrosis, inflammatory infiltrates, and glomerular and tubular changes progresses with age. Blood urea nitrogen and creatinine increase, and Glis2(mut) mice develop proteinuria and ultimately die prematurely of renal failure. A comparison of the gene expression profiles of kidneys from 25-day-old/60-day-old WT and Glis2(mut) mice by microarray analysis showed increased expressions of many genes involved in immune responses/inflammation and fibrosis/tissue remodeling in kidneys of Glis2(mut) mice, including several cytokines and adhesion and extracellular matrix proteins. Our data demonstrate that a deficiency in Glis2 expression leads to tubular atrophy and progressive fibrosis, similar to nephronophthisis, that ultimately results in renal failure. Our study indicates that Glis2 plays a critical role in the maintenance of normal kidney architecture and functions.

  1. Krüppel-Like Zinc Finger Protein Glis2 Is Essential for the Maintenance of Normal Renal Functions▿ †

    PubMed Central

    Kim, Yong-Sik; Kang, Hong Soon; Herbert, Ronald; Beak, Ju Youn; Collins, Jennifer B.; Grissom, Sherry F.; Jetten, Anton M.

    2008-01-01

    To obtain insight into the physiological functions of the Krüppel-like zinc finger protein Gli-similar 2 (Glis2), mice deficient in Glis2 expression were generated. Glis2 mutant (Glis2mut) mice exhibit significantly shorter life spans than do littermate wild-type (WT) mice due to the development of progressive chronic kidney disease with features resembling nephronophthisis. Glis2mut mice develop severe renal atrophy involving increased cell death and basement membrane thickening in the proximal convoluted tubules. This development is accompanied by infiltration of lymphocytic inflammatory cells and interstitial/glomerular fibrosis. The severity of the fibrosis, inflammatory infiltrates, and glomerular and tubular changes progresses with age. Blood urea nitrogen and creatinine increase, and Glis2mut mice develop proteinuria and ultimately die prematurely of renal failure. A comparison of the gene expression profiles of kidneys from 25-day-old/60-day-old WT and Glis2mut mice by microarray analysis showed increased expressions of many genes involved in immune responses/inflammation and fibrosis/tissue remodeling in kidneys of Glis2mut mice, including several cytokines and adhesion and extracellular matrix proteins. Our data demonstrate that a deficiency in Glis2 expression leads to tubular atrophy and progressive fibrosis, similar to nephronophthisis, that ultimately results in renal failure. Our study indicates that Glis2 plays a critical role in the maintenance of normal kidney architecture and functions. PMID:18227149

  2. Npl3, a new link between RNA-binding proteins and the maintenance of genome integrity.

    PubMed

    Santos-Pereira, José M; Herrero, Ana B; Moreno, Sergio; Aguilera, Andrés

    2014-01-01

    The mRNA is co-transcriptionally bound by a number of RNA-binding proteins (RBPs) that contribute to its processing and formation of an export-competent messenger ribonucleoprotein particle (mRNP). In the last few years, increasing evidence suggests that RBPs play a key role in preventing transcription-associated genome instability. Part of this instability is mediated by the accumulation of co-transcriptional R loops, which may impair replication fork (RF) progression due to collisions between transcription and replication machineries. In addition, some RBPs have been implicated in DNA repair and/or the DNA damage response (DDR). Recently, the Npl3 protein, one of the most abundant heterogeneous nuclear ribonucleoproteins (hnRNPs) in yeast, has been shown to prevent transcription-associated genome instability and accumulation of RF obstacles, partially associated with R-loop formation. Interestingly, Npl3 seems to have additional functions in DNA repair, and npl3∆ mutants are highly sensitive to genotoxic agents, such as the antitumor drug trabectedin. Here we discuss the role of Npl3 in particular, and RBPs in general, in the connection of transcription with replication and genome instability, and its effect on the DDR.

  3. Npl3, a new link between RNA-binding proteins and the maintenance of genome integrity

    PubMed Central

    Santos-Pereira, José M; Herrero, Ana B; Moreno, Sergio; Aguilera, Andrés

    2014-01-01

    The mRNA is co-transcriptionally bound by a number of RNA-binding proteins (RBPs) that contribute to its processing and formation of an export-competent messenger ribonucleoprotein particle (mRNP). In the last few years, increasing evidence suggests that RBPs play a key role in preventing transcription-associated genome instability. Part of this instability is mediated by the accumulation of co-transcriptional R loops, which may impair replication fork (RF) progression due to collisions between transcription and replication machineries. In addition, some RBPs have been implicated in DNA repair and/or the DNA damage response (DDR). Recently, the Npl3 protein, one of the most abundant heterogeneous nuclear ribonucleoproteins (hnRNPs) in yeast, has been shown to prevent transcription-associated genome instability and accumulation of RF obstacles, partially associated with R-loop formation. Interestingly, Npl3 seems to have additional functions in DNA repair, and npl3∆ mutants are highly sensitive to genotoxic agents, such as the antitumor drug trabectedin. Here we discuss the role of Npl3 in particular, and RBPs in general, in the connection of transcription with replication and genome instability, and its effect on the DDR. PMID:24694687

  4. Maintenance of asymmetric cellular localization of an auxin transport protein through interaction with the actin cytoskeleton

    NASA Technical Reports Server (NTRS)

    Muday, G. K.

    2000-01-01

    In shoots, polar auxin transport is basipetal (that is, from the shoot apex toward the base) and is driven by the basal localization of the auxin efflux carrier complex. The focus of this article is to summarize the experiments that have examined how the asymmetric distribution of this protein complex is controlled and the significance of this polar distribution. Experimental evidence suggests that asymmetries in the auxin efflux carrier may be established through localized secretion of Golgi vesicles, whereas an attachment of a subunit of the efflux carrier to the actin cytoskeleton may maintain this localization. In addition, the idea that this localization of the efflux carrier may control both the polarity of auxin movement and more globally regulate developmental polarity is explored. Finally, evidence indicating that the gravity vector controls auxin transport polarity is summarized and possible mechanisms for the environmentally induced changes in auxin transport polarity are discussed.

  5. Gcn5 and SAGA Regulate Shelterin Protein Turnover and Telomere Maintenance

    PubMed Central

    Atanassov, Boyko S.; Evrard, Yvonne A.; Multani, Asha S.; Zhang, Zhijing; Tora, László; Devys, Didier; Chang, Sandy; Dent, Sharon Y.R.

    2009-01-01

    SUMMARY Histone acetyltransferases (HATs) play important roles in gene regulation and DNA repair by influencing the accessibility of chromatin to transcription factors and repair proteins. Here we show that deletion of Gcn5 leads to telomere dysfunction in mouse and human cells. Biochemical studies reveal that depletion of Gcn5 or ubiquitin specific protease 22 (Usp22), which is another bona fide component of the Gcn5-containing SAGA complex, increases ubiquitination and turnover of TRF1, a primary component of the telomeric shelterin complex. Inhibition of the proteasome or over expression of USP22 opposes this effect. The USP22 deubiquitinating module requires association with SAGA complexes for activity, and we find that depletion of Gcn5 compromises this association in mammalian cells. Thus, our results indicate that Gcn5 regulates TRF1 levels through effects on Usp22 activity and SAGA integrity. PMID:19683498

  6. Maintenance of asymmetric cellular localization of an auxin transport protein through interaction with the actin cytoskeleton

    NASA Technical Reports Server (NTRS)

    Muday, G. K.

    2000-01-01

    In shoots, polar auxin transport is basipetal (that is, from the shoot apex toward the base) and is driven by the basal localization of the auxin efflux carrier complex. The focus of this article is to summarize the experiments that have examined how the asymmetric distribution of this protein complex is controlled and the significance of this polar distribution. Experimental evidence suggests that asymmetries in the auxin efflux carrier may be established through localized secretion of Golgi vesicles, whereas an attachment of a subunit of the efflux carrier to the actin cytoskeleton may maintain this localization. In addition, the idea that this localization of the efflux carrier may control both the polarity of auxin movement and more globally regulate developmental polarity is explored. Finally, evidence indicating that the gravity vector controls auxin transport polarity is summarized and possible mechanisms for the environmentally induced changes in auxin transport polarity are discussed.

  7. Transmembrane protein CD9 is glioblastoma biomarker, relevant for maintenance of glioblastoma stem cells

    PubMed Central

    Podergajs, Neža; Motaln, Helena; Rajčević, Uroš; Verbovšek, Urška; Koršič, Marjan; Obad, Nina; Espedal, Heidi; Vittori, Miloš; Herold-Mende, Christel; Miletic, Hrvoje; Bjerkvig, Rolf; Turnšek, Tamara Lah

    2016-01-01

    The cancer stem cell model suggests that glioblastomas contain a subpopulation of stem-like tumor cells that reproduce themselves to sustain tumor growth. Targeting these cells thus represents a novel treatment strategy and therefore more specific markers that characterize glioblastoma stem cells need to be identified. In the present study, we performed transcriptomic analysis of glioblastoma tissues compared to normal brain tissues revealing sensible up-regulation of CD9 gene. CD9 encodes the transmembrane protein tetraspanin which is involved in tumor cell invasion, apoptosis and resistance to chemotherapy. Using the public REMBRANDT database for brain tumors, we confirmed the prognostic value of CD9, whereby a more than two fold up-regulation correlates with shorter patient survival. We validated CD9 gene and protein expression showing selective up-regulation in glioblastoma stem cells isolated from primary biopsies and in primary organotypic glioblastoma spheroids as well as in U87-MG and U373 glioblastoma cell lines. In contrast, no or low CD9 gene expression was observed in normal human astrocytes, normal brain tissue and neural stem cells. CD9 silencing in three CD133+ glioblastoma cell lines (NCH644, NCH421k and NCH660h) led to decreased cell proliferation, survival, invasion, and self-renewal ability, and altered expression of the stem-cell markers CD133, nestin and SOX2. Moreover, CD9-silenced glioblastoma stem cells showed altered activation patterns of the Akt, MapK and Stat3 signaling transducers. Orthotopic xenotransplantation of CD9-silenced glioblastoma stem cells into nude rats promoted prolonged survival. Therefore, CD9 should be further evaluated as a target for glioblastoma treatment. PMID:26573230

  8. Transmembrane protein CD9 is glioblastoma biomarker, relevant for maintenance of glioblastoma stem cells.

    PubMed

    Podergajs, Neža; Motaln, Helena; Rajčević, Uroš; Verbovšek, Urška; Koršič, Marjan; Obad, Nina; Espedal, Heidi; Vittori, Miloš; Herold-Mende, Christel; Miletic, Hrvoje; Bjerkvig, Rolf; Turnšek, Tamara Lah

    2016-01-05

    The cancer stem cell model suggests that glioblastomas contain a subpopulation of stem-like tumor cells that reproduce themselves to sustain tumor growth. Targeting these cells thus represents a novel treatment strategy and therefore more specific markers that characterize glioblastoma stem cells need to be identified. In the present study, we performed transcriptomic analysis of glioblastoma tissues compared to normal brain tissues revealing sensible up-regulation of CD9 gene. CD9 encodes the transmembrane protein tetraspanin which is involved in tumor cell invasion, apoptosis and resistance to chemotherapy. Using the public REMBRANDT database for brain tumors, we confirmed the prognostic value of CD9, whereby a more than two fold up-regulation correlates with shorter patient survival. We validated CD9 gene and protein expression showing selective up-regulation in glioblastoma stem cells isolated from primary biopsies and in primary organotypic glioblastoma spheroids as well as in U87-MG and U373 glioblastoma cell lines. In contrast, no or low CD9 gene expression was observed in normal human astrocytes, normal brain tissue and neural stem cells. CD9 silencing in three CD133+ glioblastoma cell lines (NCH644, NCH421k and NCH660h) led to decreased cell proliferation, survival, invasion, and self-renewal ability, and altered expression of the stem-cell markers CD133, nestin and SOX2. Moreover, CD9-silenced glioblastoma stem cells showed altered activation patterns of the Akt, MapK and Stat3 signaling transducers. Orthotopic xenotransplantation of CD9-silenced glioblastoma stem cells into nude rats promoted prolonged survival. Therefore, CD9 should be further evaluated as a target for glioblastoma treatment.

  9. Immunohistochemical expression of minichromosome maintenance complex protein 2 predicts biochemical recurrence in prostate cancer: a tissue microarray and digital imaging analysis-based study of 428 cases.

    PubMed

    Toubaji, Antoun; Sutcliffe, Siobhan; Chaux, Alcides; Lecksell, Kristen; Hicks, Jessica; De Marzo, Angelo M; Platz, Elizabeth A; Netto, George J

    2012-11-01

    Prostate cancer remains a major health problem in the United States. Established clinicopathologic parameters such as Gleason score, T stage, and prostate-specific antigen levels are currently the guiding tools for prognostication and disease management. The addition of biomarkers could increase the accuracy of these parameters for predicting disease progression, response to therapy, and survival. In this regard, the goal of this study was to evaluate minichromosome maintenance complex protein 2 and Ki-67 immunohistochemical expression as predictors of outcome in prostate cancer. For this purpose, 11 tissue microarrays were constructed using tumor and nontumor samples from 428 patients. Patients were divided into short-term (mean, 2.9 years) and long-term (mean, 14.1 years) follow-up groups. End points were biochemical recurrence for the short-term follow-up group and prostate cancer-related death for the long-term follow-up group. All men in the long-term follow-up group had biochemical recurrence at the time of recruitment. Expression of both markers was higher in tumor than in nontumor glands. Percentage of minichromosome maintenance complex protein 2 was associated with Gleason score in both groups. Percentage of Ki-67 was associated with Gleason score and pathologic stage only in the short-term follow-up group. Higher minichromosome maintenance complex protein 2 percentages were associated with biochemical recurrence in the short-term follow-up group. In the long-term follow-up group, neither minichromosome maintenance complex protein 2 nor Ki-67 levels predicted prostate cancer death. In conclusion, our results suggest that in patients treated by radical prostatectomy for clinically localized prostate cancer, immunohistochemistry for minichromosome maintenance complex protein 2 expression could be used to predict biochemical recurrence, independent of other known clinicopathologic factors.

  10. The effect of a low-fat, high-protein or high-carbohydrate ad libitum diet on weight loss maintenance and metabolic risk factors.

    PubMed

    Claessens, M; van Baak, M A; Monsheimer, S; Saris, W H M

    2009-03-01

    High-protein (HP) diets are often advocated for weight reduction and weight loss maintenance. The aim was to compare the effect of low-fat, high-carbohydrate (HC) and low-fat, HP ad libitum diets on weight maintenance after weight loss induced by a very low-calorie diet, and on metabolic and cardiovascular risk factors in healthy obese subjects. Forty-eight subjects completed the study that consisted of an energy restriction period of 5-6 weeks followed by a weight maintenance period of 12 weeks. During weight maintenance subjects received maltodextrin (HC group) or protein (HP group) (casein (HPC subgroup) or whey (HPW subgroup)) supplements (2 x 25 g per day), respectively and consumed a low-fat diet. Subjects in the HP diet group showed significantly better weight maintenance after weight loss (2.3 kg difference, P=0.04) and fat mass reduction (2.2 kg difference, P=0.02) than subjects in the HC group. Triglyceride (0.6 mM difference, P=0.01) and glucagon (9.6 pg ml(-1) difference, P=0.02) concentrations increased more in the HC diet group, while glucose (0.3 mM difference, P=0.02) concentration increased more in the HP diet group. Changes in total cholesterol, low-density lipoprotein-cholesterol, high-density lipoprotein-cholesterol, insulin, HOMAir index, HbA1c, leptin and adiponectin concentrations did not differ between the diets. No differences were found between the casein- or whey-supplemented HP groups. These results show that low-fat, high-casein or whey protein weight maintenance diets are more effective for weight control than low-fat, HC diets and do not adversely affect metabolic and cardiovascular risk factors in weight-reduced moderately obese subjects without metabolic or cardiovascular complications.

  11. The E1 Protein of Human Papillomavirus Type 16 Is Dispensable for Maintenance Replication of the Viral Genome

    PubMed Central

    Egawa, Nagayasu; Nakahara, Tomomi; Ohno, Shin-ichi; Narisawa-Saito, Mako; Yugawa, Takashi; Fujita, Masatoshi; Yamato, Kenji; Natori, Yukikazu

    2012-01-01

    Papillomavirus genomes are thought to be amplified to about 100 copies per cell soon after infection, maintained constant at this level in basal cells, and amplified for viral production upon keratinocyte differentiation. To determine the requirement for E1 in viral DNA replication at different stages, an E1-defective mutant of the human papillomavirus 16 (HPV16) genome featuring a translation termination mutation in the E1 gene was used. The ability of the mutant HPV16 genome to replicate as nuclear episomes was monitored with or without exogenous expression of E1. Unlike the wild-type genome, the E1-defective HPV16 genome became established in human keratinocytes only as episomes in the presence of exogenous E1 expression. Once established, it could replicate with the same efficiency as the wild-type genome, even after the exogenous E1 was removed. However, upon calcium-induced keratinocyte differentiation, once again amplification was dependent on exogenous E1. These results demonstrate that the E1 protein is dispensable for maintenance replication but not for initial and productive replication of HPV16. PMID:22238312

  12. Post-irradiation phosphorylation of structural maintenance chromosome 1 (SMC1) is independent of the Fanconi protein pathway

    SciTech Connect

    Nahas, Shareef A.; Lai, C.-H.; Gatti, Richard A. . E-mail: rgatti@mednet.ucla.edu

    2005-03-15

    Purpose: To confirm the sensitivity of cells from patients with Fanconi anemia (FA) to ionizing radiation, and to determine whether the phosphorylation of structural maintenance chromosome 1 (SMC1) was associated with radiosensitivity, as it is in other DNA repair disorders. Methods and materials: Using lymphoblastoid cell lines from FA patients before and after exposure to ionizing radiation, the colony survival assay, radioresistant DNA synthesis, and SMC1 phosphorylation were measured. FA lymphoblastoid cell lines that had been transfected with the wild-type FANC gene were used as controls. Results: Cells from FA patients of six complementation groups were radiosensitive. Despite this, SMC1 phosphorylation was normal in each case; radioresistant DNA synthesis, a measure of S phase checkpoint integrity, was defective in FANCD2 lymphoblastoid cell lines and was corrected in FANCD2 + D2 cells. Conclusions: The data indicate that the FANC pathway proteins play a major role in the cellular responses to ionizing radiation, but not in SMC1 phosphorylation or in the S phase checkpoint of FANCD2-deficient cells. Thus, SMC1 activation is not a common denominator of radiosensitivity, as has been suggested by radiation responses of cells from ataxia-telangiectasia, Nijmegen breakage syndrome, or Mre11 deficiency patients.

  13. Serum pregnancy-associated plasma protein A correlates with inflammation and malnutrition in patients treated with maintenance hemodialysis.

    PubMed

    Mazur-Laskowska, Małgorzata; Bała-Błądzińska, Agnieszka; Zegartowska, Paulina; Dumnicka, Paulina; Ząbek-Adamska, Anna; Kapusta, Maria; Maleszka, Aleksandra; Maziarz, Barbara; Kuźniewski, Marek; Kuśnierz-Cabala, Beata

    2015-01-01

    Advanced chronic kidney disease (CKD) leads to complications such as anemia, electrolyte abnormalities, bone and mineral disorder, and malnutrition-inflammation-atherosclerosis (MIA) syndrome, that result in high cardiovascu- lar mortality. One of the biomarkers associated with inflammation and cardiovascular events is pregnancy-associated plasma protein A (PAPP-A). The aim of the study was to measure serum PAPP-A in hemodialyzed CKD patients, and to investigate its correlations with the laboratory markers of the complications. We enrolled 78 consecutive stable adult CKD patients treated with maintenance hemodialysis for median period of 60 months. PAPP-A concentrations were measured with by electrochemiluminescence immunoassay. Average serum PAPP-A in hemodialyzed patients was almost two times higher than the upper reference limit. It positively correlated with N-terminal pro-brain natriuretic peptide (NT-proBNP), serum sodium, and the markers of inflammation and malnutrition. In conclusion, serum PAPP-A seems a useful biomarker associated with cardiovascular dysfunction, inflammatory state and malnutrition in hemodialysis patients.

  14. Protein kinase C is likely to be involved in zoosporogenesis and maintenance of flagellar motility in the peronosporomycete zoospores.

    PubMed

    Islam, Md Tofazzal; von Tiedemann, Andreas; Laatsch, Hartmut

    2011-08-01

    The motility of zoospores is critical in the disease cycles of Peronosporomycetes that cause devastating diseases in plants, fishes, vertebrates, and microbes. In the course of screening for secondary metabolites, we found that ethyl acetate extracts of a marine Streptomyces sp. strain B5136 rapidly impaired the motility of zoospores of the grapevine downy mildew pathogen Plasmopara viticola at 0.1 μg/ml. The active principle in the extracts was identified as staurosporine, a known broad-spectrum inhibitor of protein kinases, including protein kinase C (PKC). In the presence of staurosporine (2 nM), zoospores moved very slowly in their axis or spun in tight circles, instead of displaying straight swimming in a helical fashion. Compounds such as K-252a, K-252b, and K-252c structurally related to staurosporine also impaired the motility of zoospores in a similar manner but at varying doses. Among the 22 known kinase inhibitors tested, the PKC inhibitor chelerythrine was the most potent to arrest the motility of zoospores at concentrations starting from 5 nM. Inhibitors that targeted kinase pathways other than PKC pathways did not practically show any activity in impairing zoospore motility. Interestingly, both staurosporine (5 nM) and chelerythrine (10 nM) also inhibited the release of zoospores from the P. viticola sporangia in a dose-dependent manner. In addition, staurosporine completely suppressed downy mildew disease in grapevine leaves at 2 μM, suggesting the potential of small-molecule PKC inhibitors for the control of peronosporomycete phytopathogens. Taken together, these results suggest that PKC is likely to be a key signaling mediator associated with zoosporogenesis and the maintenance of flagellar motility in peronosporomycete zoospores.

  15. A Role for the Twins Protein Phosphatase (PP2A-B55) in the Maintenance of Drosophila Genome Integrity.

    PubMed

    Merigliano, Chiara; Marzio, Antonio; Renda, Fioranna; Somma, Maria Patrizia; Gatti, Maurizio; Vernì, Fiammetta

    2017-03-01

    The protein phosphatase 2A (PP2A) is a conserved heterotrimeric enzyme that regulates several cellular processes including the DNA damage response and mitosis. Consistent with these functions, PP2A is mutated in many types of cancer and acts as a tumor suppressor. In mammalian cells, PP2A inhibition results in DNA double strand breaks (DSBs) and chromosome aberrations (CABs). However, the mechanisms through which PP2A prevents DNA damage are still unclear. Here, we focus on the role of the Drosophila twins (tws) gene in the maintenance of chromosome integrity; tws encodes the B regulatory subunit (B/B55) of PP2A. Mutations in tws cause high frequencies of CABs (0.5 CABs/cell) in Drosophila larval brain cells and lead to an abnormal persistence of γ-H2Av repair foci. However, mutations that disrupt the PP4 phosphatase activity impair foci dissolution but do not cause CABs, suggesting that a delayed foci regression is not clastogenic. We also show that Tws is required for activation of the G2/M DNA damage checkpoint while PP4 is required for checkpoint recovery, a result that points to a conserved function of these phosphatases from flies to humans. Mutations in the ATM-coding gene tefu are strictly epistatic to tws mutations for the CAB phenotype, suggesting that failure to dephosphorylate an ATM substrate(s) impairs DNA DSBs repair. In addition, mutations in the Ku70 gene, which do not cause CABs, completely suppress CAB formation in tws Ku70 double mutants. These results suggest the hypothesis that an improperly phosphorylated Ku70 protein can lead to DNA damage and CABs. Copyright © 2017 by the Genetics Society of America.

  16. Effect of a high-protein diet on maintenance of blood pressure levels achieved after initial weight loss: the DiOGenes randomized study.

    PubMed

    Engberink, M F; Geleijnse, J M; Bakker, S J L; Larsen, T M; Handjieva-Darlesnka, T; Kafatos, A; Martinez, J A; Pfeiffer, A F H; Kunešová, M; Jebb, S A; Holst, C; Astrup, A; Saris, W H M; Brink, E J; van Baak, M A

    2015-01-01

    Randomized trials have shown significant blood pressure (BP) reductions after increased protein compared with carbohydrate intake, but the effect on BP maintenance after initial weight loss is unclear. We examined the effect of a high-protein diet on the maintenance of reduced BP after weight loss in 420 overweight adults from the Diet, Obesity and Genes study. After an 8-week weight-loss period (>8% BW), subjects (42±6 years) were randomized to either a high-protein diet (23-28 en% protein) or a lower-protein control diet (10-15 en% protein) for 26 weeks. BMI after weight loss was 30.3±4.3 kg m(-2), BP was 118/73 mm Hg and 28 subjects (6.5%) used antihypertensive agents. Systolic BP during 26 weeks of weight maintenance dietary intervention increased in both treatment groups, but it was 2.2 mm Hg less (95% CI: -4.6 to 0.2 mm Hg, P=0.08) in the high-protein group than in the lower-protein control group. In 191 (pre)hypertensive subjects (baseline systolic BP⩾120 mm Hg), a larger difference was observed (-4.2 mm Hg (-7.7, -0.7), P=0.02). The effect was attenuated after adjustment for initial BP (-3.4 mm Hg (-6.9, -0.03), P=0.048), and after additional adjustment for weight change (-2.7 mm Hg (-6.1, 0.4), P=0.11). Adjustment for 24-h urinary excretion of sodium and potassium did not change the results. Diastolic BP yielded similar results. These findings suggest that a BP reduction after weight loss is better maintained when the intake of protein is increased at the expense of carbohydrates. This effect is partly mediated by body weight.

  17. A Randomized Clinical Trial of the Effect of Pentoxifylline on C-Reactive Protein Level and Dialysis Adequacy in End-stage Renal Disease Patients on Maintenance Hemodialysis.

    PubMed

    Soltani, Parvin; Ketabi Moghaddam, Pardis; Haghverdi, Farshid; Cheraghi, Ali

    2016-09-01

     C-reactive protein (CRP) is increased among patients on maintenance hemodialysis. Such inflammatory markers can result in protein-energy deficit syndromes and low adequacy of dialysis in these patients. This study evaluated the effect of pentoxifylline on serum CRP level and KT/V in end-stage renal disease patients on maintenance hemodialysis. This 1-month randomized, double-blind, placebo-controlled clinical trial involving 73 patients with end-stage renal disease on maintenance hemodialysis assessed the effectiveness of 400 mg/d of pentoxifylline on serum CRP level decrease and improvement of dialysis adequacy. The difference in mean serum CRP levels of the pentoxifylline and placebo groups was not significant before study. While CRP showed showed a significant increase in the placebo group after completing the interventions (P = .01), the difference was nonsignificant in the pentoxifylline group (P = .53). The difference in the mean adequacy of dialysis was not significant before the interventions between the two groups, while there was a significant increase in the pentoxifylline group (P = .01) and a nonsignificant increase in the placebo group (P = .31) after the interventions. Among patients on maintenance hemodialysis, a 1-month trial of pentoxifylline was associated with a substantial improvement of adequacy of dialysis and a significant prevention from serum CRP level increase, but not a significant reduction in the mean serum CRP level.

  18. Protein Kinase M[Zeta] Is Essential for the Induction and Maintenance of Dopamine-Induced Long-Term Potentiation in Apical CA1 Dendrites

    ERIC Educational Resources Information Center

    Navakkode, Sheeja; Sajikumar, Sreedharan; Sacktor, Todd Charlton; Frey, Julietta U.

    2010-01-01

    Dopaminergic D1/D5-receptor-mediated processes are important for certain forms of memory as well as for a cellular model of memory, hippocampal long-term potentiation (LTP) in the CA1 region of the hippocampus. D1/D5-receptor function is required for the induction of the protein synthesis-dependent maintenance of CA1-LTP (L-LTP) through activation…

  19. DomainSieve: a protein domain-based screen that led to the identification of dam-associated genes with potential link to DNA maintenance.

    PubMed

    Brézellec, Pierre; Hoebeke, Mark; Hiet, Marie-Sophie; Pasek, Sophie; Ferat, Jean-Luc

    2006-08-15

    The Dam methyltransferase (DamMT) activity, broadly distributed in association with restriction endonucleases, as part of the restriction-modification defense systems, has evolved to become intimately associated with essential biological functions in a few organisms. In Escherichia coli, DamMT is involved in multiple aspects of DNA maintenance, replication initiation, daughter chromosome segregation, DNA mismatch repair, gene expression control, etc. The participation of DamMT in such a diverse set of functions required that other genes adapted, or emerged through evolution, in response to the DamMT-induced modification of the genomic environment. One example is SeqA, a protein that senses the methylation status of the origin of replication of the chromosome to control the timing of replication initiation. Interestingly, seqA is only present in a few DamMT-specifying proteobacteria. This observation led us to hypothesize that other genes, specifying related functions, might also be found in these organisms. To test this hypothesis, we implemented a large-scale comparative genomic screen meant to identify genes specifying DNA methylation sensing domains, probably involved in DNA maintenance functions. We carried out a phylogenetic analysis of DamMT, identifying two contrasting behaviors of the protein. Based on this phylogeny, we defined precisely a set of genomes, in which the protein activity is likely to be involved in DNA maintenance functions, the 'resident' dam genomes. We defined a second set of genomes, in which DamMT is not resident. We developped a new tool, 'DomainSieve', in order to screen these two sets for protein domains that are strictly associated with 'resident' dam genomes. This approach was rewarding and generated a list of genes, among which some, at least, specify activities with clear linkage to DamMT-dependent DNA methylation and DNA maintenance. DomainSieve is implemented as a web resource and is accessible at http://stat.genopole.cnrs.fr/ds/.

  20. Protein Kinase M[Zeta] Is Essential for the Induction and Maintenance of Dopamine-Induced Long-Term Potentiation in Apical CA1 Dendrites

    ERIC Educational Resources Information Center

    Navakkode, Sheeja; Sajikumar, Sreedharan; Sacktor, Todd Charlton; Frey, Julietta U.

    2010-01-01

    Dopaminergic D1/D5-receptor-mediated processes are important for certain forms of memory as well as for a cellular model of memory, hippocampal long-term potentiation (LTP) in the CA1 region of the hippocampus. D1/D5-receptor function is required for the induction of the protein synthesis-dependent maintenance of CA1-LTP (L-LTP) through activation…

  1. Sirtinol, a Sir2 protein inhibitor, affects stem cell maintenance and root development in Arabidopsis thaliana by modulating auxin-cytokinin signaling components

    PubMed Central

    Singh, Sharmila; Singh, Alka; Yadav, Sandeep; Gautam, Vibhav; Singh, Archita; Sarkar, Ananda K.

    2017-01-01

    In Arabidopsis thaliana, besides several key transcription factors and chromatin modifiers, phytohormones auxin and cytokinin play pivotal role in shoot and root meristem maintenance, and lateral root (LR) development. Sirtinol, a chemical inhibitor of Sir2 proteins, is known to promote some auxin induced phenotypes in Arabidopsis. However, its effect on plant stem cell maintenance or organ formation remained unaddressed. Here we show that sirtinol affects meristem maintenance by altering the expression of key stem cell regulators, cell division and differentiation by modulating both auxin and cytokinin signaling in Arabidopsis thaliana. The expression of shoot stem cell niche related genes WUSCHEL (WUS) and CLAVATA3 (CLV3) was upregulated, whereas SHOOT MERISTEMLESS (STM) was downregulated in sirtinol treated seedlings. The expression level and domain of key root stem cell regulators PLETHORA (PLTs) and WUS-Related Homeobox 5 (WOX5) were altered in sirtinol treated roots. Sirtinol affects LR development by disturbing proper auxin transport and maxima formation, similar to 2,4-dichlorophenoxyacetic acid (2,4-D). Sirtinol also affects LR formation by altering cytokinin biosynthesis and signaling genes in roots. Therefore, sirtinol affects shoot and root growth, meristem maintenance and LR development by altering the expression of cytokinin-auxin signaling components, and regulators of stem cells, meristems, and LRs. PMID:28195159

  2. Memory reconsolidation and its maintenance depend on L-voltage-dependent calcium channels and CaMKII functions regulating protein turnover in the hippocampus.

    PubMed

    Da Silva, Weber Cláudio; Cardoso, Gabriela; Bonini, Juliana Sartori; Benetti, Fernando; Izquierdo, Ivan

    2013-04-16

    Immediate postretrieval bilateral blockade of long-acting voltage-dependent calcium channels (L-VDCCs), but not of glutamatergic NMDA receptors, in the dorsal CA1 region of the hippocampus hinders retention of long-term spatial memory in the Morris water maze. Immediate postretrieval bilateral inhibition of calcium/calmodulin-dependent protein kinase (CaMK) II in dorsal CA1 does not affect retention of this task 24 h later but does hinder it 5 d later. These two distinct amnesic effects are abolished if protein degradation by proteasomes is inhibited concomitantly. These results indicate that spatial memory reconsolidation depends on the functionality of L-VDCC in dorsal CA1, that maintenance of subsequent reconsolidated memory trace depends on CaMKII, and these results also suggest that the role played by both L-VDCC and CaMKII is to promote the retrieval-dependent, synaptically localized enhancement of protein synthesis necessary to counteract a retrieval-dependent, synaptic-localized enhancement of protein degradation, which has been described as underlying the characteristic labilization of the memory trace triggered by retrieval. Thus, conceivably, L-VDCC and CaMKII would enhance activity-dependent localized protein renewal, which may account for the improvement of the long-term efficiency of the synapses responsible for the maintenance of reactivated long-term spatial memory.

  3. Memory reconsolidation and its maintenance depend on L-voltage-dependent calcium channels and CaMKII functions regulating protein turnover in the hippocampus

    PubMed Central

    Da Silva, Weber Cláudio; Cardoso, Gabriela; Bonini, Juliana Sartori; Benetti, Fernando; Izquierdo, Ivan

    2013-01-01

    Immediate postretrieval bilateral blockade of long-acting voltage–dependent calcium channels (L-VDCCs), but not of glutamatergic NMDA receptors, in the dorsal CA1 region of the hippocampus hinders retention of long-term spatial memory in the Morris water maze. Immediate postretrieval bilateral inhibition of calcium/calmodulin-dependent protein kinase (CaMK) II in dorsal CA1 does not affect retention of this task 24 h later but does hinder it 5 d later. These two distinct amnesic effects are abolished if protein degradation by proteasomes is inhibited concomitantly. These results indicate that spatial memory reconsolidation depends on the functionality of L-VDCC in dorsal CA1, that maintenance of subsequent reconsolidated memory trace depends on CaMKII, and these results also suggest that the role played by both L-VDCC and CaMKII is to promote the retrieval-dependent, synaptically localized enhancement of protein synthesis necessary to counteract a retrieval-dependent, synaptic-localized enhancement of protein degradation, which has been described as underlying the characteristic labilization of the memory trace triggered by retrieval. Thus, conceivably, L-VDCC and CaMKII would enhance activity-dependent localized protein renewal, which may account for the improvement of the long-term efficiency of the synapses responsible for the maintenance of reactivated long-term spatial memory. PMID:23576750

  4. Role of protein kinase C in the induction and maintenance of serotonin-dependent enhancement of the glutamate response in isolated siphon motor neurons of Aplysia californica.

    PubMed

    Villareal, Greg; Li, Quan; Cai, Diancai; Fink, Ann E; Lim, Travis; Bougie, Joanna K; Sossin, Wayne S; Glanzman, David L

    2009-04-22

    Serotonin (5-HT) mediates learning-related facilitation of sensorimotor synapses in Aplysia californica. Under some circumstances 5-HT-dependent facilitation requires the activity of protein kinase C (PKC). One critical site of PKC's contribution to 5-HT-dependent synaptic facilitation is the presynaptic sensory neuron. Here, we provide evidence that postsynaptic PKC also contributes to synaptic facilitation. We investigated the contribution of PKC to enhancement of the glutamate-evoked potential (Glu-EP) in isolated siphon motor neurons in cell culture. A 10 min application of either 5-HT or phorbol ester, which activates PKC, produced persistent (> 50 min) enhancement of the Glu-EP. Chelerythrine and bisindolylmaleimide-1 (Bis), two inhibitors of PKC, both blocked the induction of 5-HT-dependent enhancement. An inhibitor of calpain, a calcium-dependent protease, also blocked 5-HT's effect. Interestingly, whereas chelerythrine blocked maintenance of the enhancement, Bis did not. Because Bis has greater selectivity for conventional and novel isoforms of PKC than for atypical isoforms, this result implicates an atypical isoform in the maintenance of 5-HT's effect. Although induction of enhancement of the Glu-EP requires protein synthesis (Villareal et al., 2007), we found that maintenance of the enhancement does not. Maintenance of 5-HT-dependent enhancement appears to be mediated by a PKM-type fragment generated by calpain-dependent proteolysis of atypical PKC. Together, our results suggest that 5-HT treatment triggers two phases of PKC activity within the motor neuron, an early phase that may involve conventional, novel or atypical isoforms of PKC, and a later phase that selectively involves an atypical isoform.

  5. Comparative Effects of 3 Popular Diets on Lipids, Endothelial Function and C-Reactive Protein during Weight Maintenance

    PubMed Central

    2009-01-01

    Although popular diets focus on weight loss and their favorable biochemical and physiological effects, fewer investigations have evaluated the biological impact of these diets during weight maintenance. To study this issue, three popular diets, Atkins, South Beach and Ornish were tested in a randomized and counterbalanced, crossover study between January and December 2006. Participants completed each of the three 4-week isocaloric dietary intervention phases followed by a 4-week washout period. They were weighed weekly and caloric adjustments made if weight change exceeded 1 kg. At the completion of each dietary phase, 3-day food records were analyzed, fasting blood sampled and brachial artery reactivity testing (BART) performed. Eighteen adults completed all 3 isocaloric dietary phases. During the South Beach and Ornish maintenance phase there were significant reductions in low density lipoprotein cholesterol (LDL-C) (11.8% [P=0.01], 16.6% [P=0.0006], respectively) compared to pre-diet baseline. Moreover, in contrast to the Atkins maintenance phase, significant reductions in LDL-C and apolipoprotein B levels were observed after the South Beach (P=0.003, P=0.05, repeated measures ANOVA) and Ornish maintenance phase (P=0.0004, P=0.006, repeated measures ANOVA). Brachial artery testing revealed an inverse correlation between flow-mediated vasodilatation and intake of saturated fat (r=−0.33; p=0.016). These data suggest that during weight maintenance, less favorable biological effects are observed during a simulated, high fat Atkins diet when compared to the South Beach and Ornish diet. The findings support additional study in subjects with visceral obesity and the metabolic syndrome, in whom an increased risk of coronary disease at baseline may be accentuated with chronic consumption of a diet that exhibits unfavorable effects on lipids and endothelial function. PMID:19328268

  6. Comparative effects of three popular diets on lipids, endothelial function, and C-reactive protein during weight maintenance.

    PubMed

    Miller, Michael; Beach, Valerie; Sorkin, John D; Mangano, Charles; Dobmeier, Christine; Novacic, Danica; Rhyne, Jeffrey; Vogel, Robert A

    2009-04-01

    Although popular diets focus on weight loss and their favorable biochemical and physiological effects, fewer investigations have evaluated the biological impact of these diets during weight maintenance. To study this issue, three popular diets-Atkins, South Beach, and Ornish-were tested in a randomized and counterbalanced crossover study between January and December 2006. Participants completed each of the three 4-week isocaloric dietary intervention phases followed by a 4-week washout period. They were weighed weekly and caloric adjustments made if weight change exceeded 1 kg. At the completion of each dietary phase, 3-day food records were analyzed, fasting blood sampled, and brachial artery reactivity testing performed. Eighteen adults completed all three isocaloric dietary phases. During the South Beach and Ornish maintenance phase, there were significant reductions in low-density lipoprotein cholesterol (11.8%; P=0.01, 16.6%; P=0.0006, respectively) compared to prediet baseline. In addition, in contrast to the Atkins maintenance phase, significant reductions in low-density lipoprotein cholesterol and apolipoprotein B levels were observed after the South Beach (P=0.003, P=0.05; repeated measures analyses of variance) and Ornish maintenance phases (P=0.0004, P=0.006, repeated measures analyses of variance). Brachial artery testing revealed an inverse correlation between flow-mediated vasodilatation and intake of saturated fat (r=-0.33; P=0.016). These data suggest that during weight maintenance, less favorable biological effects are observed during a simulated, high-fat Atkins diet when compared to the South Beach and Ornish diet. The findings support additional study in subjects with visceral obesity and the metabolic syndrome, in whom an increased risk of coronary disease at baseline may be accentuated with chronic consumption of a diet that exhibits unfavorable effects on lipids and endothelial function.

  7. Preventative Maintenance.

    ERIC Educational Resources Information Center

    Migliorino, James

    Boards of education must be convinced that spending money up front for preventive maintenance will, in the long run, save districts' tax dollars. A good program of preventive maintenance can minimize disruption of service; reduce repair costs, energy consumption, and overtime; improve labor productivity and system equipment reliability; handle…

  8. Preventative Maintenance.

    ERIC Educational Resources Information Center

    Migliorino, James

    Boards of education must be convinced that spending money up front for preventive maintenance will, in the long run, save districts' tax dollars. A good program of preventive maintenance can minimize disruption of service; reduce repair costs, energy consumption, and overtime; improve labor productivity and system equipment reliability; handle…

  9. Maintenance Downtime

    Atmospheric Science Data Center

    2013-07-10

    ... will be unavailable March 5, 2013 8:00 am to 5:00 pm due to database maintenance. Date(s):  Tuesday, March 5, 2013 ... will be unavailable March 5, 2013 8:00 am to 5:00 pm due to database maintenance. ...

  10. Maintenance Budgeting.

    ERIC Educational Resources Information Center

    Smith, J. McCree

    Three methods for the preparation of maintenance budgets are discussed--(1) a traditional method, inconclusive and obsolete, based on gross square footage, (2) the formula approach method based on building classification (wood-frame, masonry-wood, masonry-concrete) with maintenance cost factors for each type plus custodial service rates by type of…

  11. Software Maintenance.

    ERIC Educational Resources Information Center

    Cannon, Glenn; Jobe, Holly

    Proper cleaning and storage of audiovisual aids is outlined in this brief guide. Materials and equipment needed for first line maintenance are listed, as well as maintenance procedures for records, audio and video tape, film, filmstrips, slides, realia, models, prints, graphics, maps, and overhead transparencies. A 15-item quiz on software…

  12. Comparison of Subjective Global Assessment and Protein Energy Wasting Score to Nutrition Evaluations Conducted by Registered Dietitian Nutritionists in Identifying Protein Energy Wasting Risk in Maintenance Hemodialysis Patients.

    PubMed

    Sum, Simon Siu-Man; Marcus, Andrea F; Blair, Debra; Olejnik, Laura A; Cao, Joyce; Parrott, J Scott; Peters, Emily N; Hand, Rosa K; Byham-Gray, Laura D

    2017-09-01

    To compare the 7-point subjective global assessment (SGA) and the protein energy wasting (PEW) score with nutrition evaluations conducted by registered dietitian nutritionists in identifying PEW risk in stage 5 chronic kidney disease patients on maintenance hemodialysis. This study is a secondary analysis of a cross-sectional study entitled "Development and Validation of a Predictive energy Equation in Hemodialysis". PEW risk identified by the 7-point SGA and the PEW score was compared against the nutrition evaluations conducted by registered dietitian nutritionists through data examination from the original study (reference standard). A total of 133 patients were included for the analysis. The sensitivity, specificity, positive and negative predictive value (PPV and NPV), positive and negative likelihood ratio (PLR and NLR) of both scoring tools were calculated when compared against the reference standard. The patients were predominately African American (n = 112, 84.2%), non-Hispanic (n = 101, 75.9%), and male (n = 80, 60.2%). Both the 7-point SGA (sensitivity = 78.6%, specificity = 59.1%, PPV = 33.9%, NPV = 91.2%, PLR = 1.9, and NLR = 0.4) and the PEW score (sensitivity = 100%, specificity = 28.6%, PPV = 27.2%, NPV = 100%, PLR = 1.4, and NLR = 0) were more sensitive than specific in identifying PEW risk. The 7-point SGA may miss 21.4% patients having PEW and falsely identify 40.9% of patients who do not have PEW. The PEW score can identify PEW risk in all patients, but 71.4% of patients identified may not have PEW risk. Both the 7-point SGA and the PEW score could identify PEW risk. The 7-point SGA was more specific, and the PEW score was more sensitive. Both scoring tools were found to be clinically confident in identifying patients who were actually not at PEW risk. Copyright © 2017 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

  13. Green tea catechin plus caffeine supplementation to a high-protein diet has no additional effect on body weight maintenance after weight loss.

    PubMed

    Hursel, Rick; Westerterp-Plantenga, Margriet S

    2009-03-01

    Green tea (epigallocatechin gallate + caffeine) and protein each were shown to improve body weight maintenance after weight loss. We investigated the effect of a green tea-caffeine mixture added to a high-protein (HP) diet on weight maintenance (WM) after body weight loss in moderately obese subjects. A randomized, placebo-controlled, double-blind parallel trial was conducted in 80 overweight and moderately obese subjects [age (mean +/- SD): 44 +/- 2 y; body mass index (BMI; in kg/m(2)): 29.6 +/- 2.0] matched for sex, age, BMI, height, body mass, and with a habitually low caffeine intake. A very-low-energy diet intervention during 4 wk was followed by 3 mo of WM; during the WM period, the subjects received a green tea-caffeine mixture (270 mg epigallocatechin gallate + 150 mg caffeine/d) or placebo, both in addition to an adequate protein (AP) diet (50-60 g protein/d) or an HP diet (100-120 g protein/d). Subjects lost 7.0 +/- 1.6 kg, or 8.2 +/- 2.0%, body weight (P < 0.001). During the WM phase, WM, resting energy expenditure, and fat-free mass (FFM) increased relatively in both the HP groups and in the AP + green tea-caffeine mixture group (P < 0.05), whereas respiratory quotient and body fat mass decreased, all compared with the AP + placebo group. Satiety increased only in both HP groups (P < 0.05). The green tea-caffeine mixture was only effective with the AP diet. The green tea-caffeine mixture, as well as the HP diet, improved WM independently through thermogenesis, fat oxidation, sparing FFM, and, for the HP diet, satiety; a possible synergistic effect failed to appear.

  14. Arp2/3 promotes junction formation and maintenance in the Caenorhabditis elegans intestine by regulating membrane association of apical proteins

    PubMed Central

    Bernadskaya, Yelena Y.; Patel, Falshruti B.; Hsu, Hsiao-Ting; Soto, Martha C.

    2011-01-01

    It has been proposed that Arp2/3, which promotes nucleation of branched actin, is needed for epithelial junction initiation but is less important as junctions mature. We focus here on how Arp2/3 contributes to the Caenorhabditis elegans intestinal epithelium and find important roles for Arp2/3 in the maturation and maintenance of junctions in embryos and adults. Electron microscope studies show that embryos depleted of Arp2/3 form apical actin-rich microvilli and electron-dense apical junctions. However, whereas apical/basal polarity initiates, apical maturation is defective, including decreased apical F-actin enrichment, aberrant lumen morphology, and reduced accumulation of some apical junctional proteins, including DLG-1. Depletion of Arp2/3 in adult animals leads to similar intestinal defects. The DLG-1/AJM-1 apical junction proteins, and the ezrin–radixin–moesin homologue ERM-1, a protein that connects F-actin to membranes, are required along with Arp2/3 for apical F-actin enrichment in embryos, whereas cadherin junction proteins are not. Arp2/3 affects the subcellular distribution of DLG-1 and ERM-1. Loss of Arp2/3 shifts both ERM-1 and DLG-1 from pellet fractions to supernatant fractions, suggesting a role for Arp2/3 in the distribution of membrane-associated proteins. Thus, Arp2/3 is required as junctions mature to maintain apical proteins associated with the correct membranes. PMID:21697505

  15. Protein kinase Mζ is essential for the induction and maintenance of dopamine-induced long-term potentiation in apical CA1 dendrites

    PubMed Central

    Navakkode, Sheeja; Sajikumar, Sreedharan; Sacktor, Todd Charlton; Frey, Julietta U.

    2010-01-01

    Dopaminergic D1/D5-receptor-mediated processes are important for certain forms of memory as well as for a cellular model of memory, hippocampal long-term potentiation (LTP) in the CA1 region of the hippocampus. D1/D5-receptor function is required for the induction of the protein synthesis-dependent maintenance of CA1-LTP (L-LTP) through activation of the cAMP/PKA-pathway. In earlier studies we had reported a synergistic interaction of D1/D5-receptor function and N-methyl-D-aspartate (NMDA)-receptors for L-LTP. Furthermore, we have found the requirement of the atypical protein kinase C isoform, protein kinase Mζ (PKMζ) for conventional electrically induced L-LTP, in which PKMζ has been identified as a LTP-specific plasticity-related protein (PRP) in apical CA1-dendrites. Here, we investigated whether the dopaminergic pathway activates PKMζ. We found that application of dopamine (DA) evokes a protein synthesis-dependent LTP that requires synergistic NMDA-receptor activation and protein synthesis in apical CA1-dendrites. We identified PKMζ as a DA-induced PRP, which exerted its action at activated synaptic inputs by processes of synaptic tagging. PMID:21084457

  16. Topoisomerase II-associated protein PAT1H1 is involved in the root stem cell niche maintenance in Arabidopsis thaliana.

    PubMed

    Yu, Qianqian; Liu, Jiajia; Zheng, Huihui; Jia, Yuebin; Tian, Huiyu; Ding, Zhaojun

    2016-06-01

    PAT1H1, one of the homologues of Topoisomerase II-associated protein, is involved in the maintenance of root stem cell niche through the interaction with NINJA. The root stem cell niche, which possesses four mitotically inactive quiescent cells (QC) and the surrounding mitotically active stem cells, is critical for root development in Arabidopsis thaliana. However, the molecular regulation of the maintenance of root stem cell niche identity is still not fully understood. Here we show that one of the homologues of Topoisomerase II-associated protein, here named as PAT1H1, could regulate root stem cell niche identity. The pat1h1 mutant showed higher frequency of QC cell division and root distal stem cell (DSC) differentiation. With a high expression in roots, PAT1H1 was found to interact with the jasmonic acid (JA) signalling negative regulator Novel Interactor of JAZ (NINJA) and thus regulate root DSC niche identity. Consistent with the active QC cell division, which rarely occurs in wild-type controls, the pat1h1 mutant displayed higher expression of CYCB1 in the root stem cell niche. Together our data reveals that PAT1H1 maintains root stem cell niche stability through the interaction with NINJA and the regulation of cell division.

  17. Chromatin Association of Human Origin Recognition Complex, Cdc6, and Minichromosome Maintenance Proteins during the Cell Cycle: Assembly of Prereplication Complexes in Late Mitosis

    PubMed Central

    Méndez, Juan; Stillman, Bruce

    2000-01-01

    Evidence obtained from studies with yeast and Xenopus indicate that the initiation of DNA replication is a multistep process. The origin recognition complex (ORC), Cdc6p, and minichromosome maintenance (MCM) proteins are required for establishing prereplication complexes, upon which initiation is triggered by the activation of cyclin-dependent kinases and the Dbf4p-dependent kinase Cdc7p. The identification of human homologues of these replication proteins allows investigation of S-phase regulation in mammalian cells. Using centrifugal elutriation of several human cell lines, we demonstrate that whereas human Orc2 (hOrc2p) and hMcm proteins are present throughout the cell cycle, hCdc6p levels vary, being very low in early G1 and accumulating until cells enter mitosis. hCdc6p can be polyubiquitinated in vivo, and it is stabilized by proteasome inhibitors. Similar to the case for hOrc2p, a significant fraction of hCdc6p is present on chromatin throughout the cell cycle, whereas hMcm proteins alternate between soluble and chromatin-bound forms. Loading of hMcm proteins onto chromatin occurs in late mitosis concomitant with the destruction of cyclin B, indicating that the mitotic kinase activity inhibits prereplication complex formation in human cells. PMID:11046155

  18. The Symmetrical Structure of Structural Maintenance of Chromosomes (SMC) and MukB Proteins: Long, Antiparallel Coiled Coils, Folded at a Flexible Hinge

    PubMed Central

    Melby, Thomas E.; Ciampaglio, Charles N.; Briscoe, Gina; Erickson, Harold P.

    1998-01-01

    Structural maintenance of chromosomes (SMC) proteins function in chromosome condensation and several other aspects of DNA processing. They are large proteins characterized by an NH2-terminal nucleotide triphosphate (NTP)-binding domain, two long segments of coiled coil separated by a hinge, and a COOH-terminal domain. Here, we have visualized by EM the SMC protein from Bacillus subtilis (BsSMC) and MukB from Escherichia coli, which we argue is a divergent SMC protein. Both BsSMC and MukB show two thin rods with globular domains at the ends emerging from the hinge. The hinge appears to be quite flexible: the arms can open up to 180°, separating the terminal domains by 100 nm, or close to near 0°, bringing the terminal globular domains together. A surprising observation is that the ∼300–amino acid–long coiled coils are in an antiparallel arrangement. Known coiled coils are almost all parallel, and the longest antiparallel coiled coils known previously are 35–45 amino acids long. This antiparallel arrangement produces a symmetrical molecule with both an NH2- and a COOH-terminal domain at each end. The SMC molecule therefore has two complete and identical functional domains at the ends of the long arms. The bifunctional symmetry and a possible scissoring action at the hinge should provide unique biomechanical properties to the SMC proteins. PMID:9744887

  19. AGAMOUS Terminates Floral Stem Cell Maintenance in Arabidopsis by Directly Repressing WUSCHEL through Recruitment of Polycomb Group Proteins[W

    PubMed Central

    Liu, Xigang; Kim, Yun Ju; Müller, Ralf; Yumul, Rae Eden; Liu, Chunyan; Pan, Yanyun; Cao, Xiaofeng; Goodrich, Justin; Chen, Xuemei

    2011-01-01

    Floral stem cells produce a defined number of floral organs before ceasing to be maintained as stem cells. Therefore, floral stem cells offer an ideal model to study the temporal control of stem cell maintenance within a developmental context. AGAMOUS (AG), a MADS domain transcription factor essential for the termination of floral stem cell fate, has long been thought to repress the stem cell maintenance gene WUSCHEL (WUS) indirectly. Here, we uncover a role of Polycomb Group (PcG) genes in the temporally precise repression of WUS expression and termination of floral stem cell fate. We show that AG directly represses WUS expression by binding to the WUS locus and recruiting, directly or indirectly, PcG that methylates histone H3 Lys-27 at WUS. We also show that PcG acts downstream of AG and probably in parallel with the known AG target KNUCKLES to terminate floral stem cell fate. Our studies identify core components of the network governing the temporal program of floral stem cells. PMID:22028461

  20. Small G protein Rac GTPases regulate the maintenance of glioblastoma stem-like cells in vitro and in vivo.

    PubMed

    Lai, Yun-Ju; Tsai, Jui-Cheng; Tseng, Ying-Ting; Wu, Meng-Shih; Liu, Wen-Shan; Lam, Hoi-Ian; Yu, Jei-Hwa; Nozell, Susan E; Benveniste, Etty N

    2017-03-14

    Glioblastoma is the most common and aggressive malignant brain tumor in adults. The existence of glioblastoma stem cells (GSCs) or stem-like cells (stemloids) may account for its invasiveness and high recurrence. Rac proteins belong to the Rho small GTPase subfamily which regulates cell movement, proliferation, and survival. To investigate whether Rac proteins can serve as therapeutic targets for glioblastoma, especially for GSCs or stemloids, we examined the potential roles of Rac1, Rac2 and Rac3 on the properties of tumorspheres derived from glioblastoma cell lines. Tumorspheres are thought to be glioblastoma stem-like cells. We showed that Rac proteins promote the STAT3 and ERK activation and enhance cell proliferation and colony formation of glioblastoma stem-like cells. Knockdown of Rac proteins reduces the expression of GSC markers, such as CD133 and Sox2. The in vivo effects of Rac proteins in glioblastoma were further studied in zebrafish and in the mouse xenotransplantation model. Knocking-down Rac proteins abolished the angiogenesis effect induced by the injected tumorspheres in zebrafish model. In the CD133+-U373-tumorsphere xenotransplanted mouse model, suppression of Rac proteins decreased the incidence of tumor formation and inhibited the tumor growth. Moreover, knockdown of Rac proteins reduced the sphere forming efficiency of cells derived from these tumors. In conclusion, not only Rac1 but also Rac2 and 3 are important for glioblastoma tumorigenesis and can serve as the potential therapeutic targets against glioblastoma and its stem-like cells.

  1. Maintenance of energy expenditure on high-protein vs. high-carbohydrate diets at a constant body weight may prevent a positive energy balance.

    PubMed

    Martens, E A; Gonnissen, H K; Gatta-Cherifi, B; Janssens, P L; Westerterp-Plantenga, M S

    2015-10-01

    Relatively high-protein diets are effective for body weight loss, and subsequent weight maintenance, yet it remains to be shown whether these diets would prevent a positive energy balance. Therefore, high-protein diet studies at a constant body weight are necessary. The objective was to determine fullness, energy expenditure, and macronutrient balances on a high-protein low-carbohydrate (HPLC) diet compared with a high-carbohydrate low-protein (HCLP) diet at a constant body weight, and to assess whether effects are transient or sustained after 12 weeks. A randomized parallel study was performed in 14 men and 18 women [mean ± SD age: 24 ± 5 y; BMI (in kg/m(2)): 22.8 ± 2.0] on diets containing 30/35/35 (HPLC) or 5/60/35 (HCLP) % of energy from protein/carbohydrate/fat. Significant interactions between dietary intervention and time on total energy expenditure (TEE) (P = 0.013), sleeping metabolic rate (SMR) (P = 0.040), and diet-induced thermogenesis (DIT) (P = 0.027) appeared from baseline to wk 12. TEE was maintained in the HPLC diet group, while it significantly decreased throughout the intervention period in the HCLP diet group (wk 1: P = 0.002; wk 12: P = 0.001). Energy balance was maintained in the HPLC diet group, and became positive in the HCLP diet group at wk 12 (P = 0.008). Protein balance varied directly according to the amount of protein in the diet, and diverged significantly between the diets (P = 0.001). Fullness ratings were significantly higher in the HPLC vs. the HCLP diet group at wk 1 (P = 0.034), but not at wk 12. Maintenance of energy expenditure on HPLC vs. HCLP diets at a constant body weight may prevent development of a positive energy balance, despite transiently higher fullness. The study was registered on clinicaltrials.gov with Identifier: NCT01551238. Copyright © 2014 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

  2. A Small Zinc Finger Thylakoid Protein Plays a Role in Maintenance of Photosystem II in Arabidopsis thaliana[W][OA

    PubMed Central

    Lu, Yan; Hall, David A.; Last, Robert L.

    2011-01-01

    This work identifies LOW QUANTUM YIELD OF PHOTOSYSTEM II1 (LQY1), a Zn finger protein that shows disulfide isomerase activity, interacts with the photosystem II (PSII) core complex, and may act in repair of photodamaged PSII complexes. Two mutants of an unannotated small Zn finger containing a thylakoid membrane protein of Arabidopsis thaliana (At1g75690; LQY1) were found to have a lower quantum yield of PSII photochemistry and reduced PSII electron transport rate following high-light treatment. The mutants dissipate more excess excitation energy via nonphotochemical pathways than wild type, and they also display elevated accumulation of reactive oxygen species under high light. After high-light treatment, the mutants have less PSII–light-harvesting complex II supercomplex than wild-type plants. Analysis of thylakoid membrane protein complexes showed that wild-type LQY1 protein comigrates with the PSII core monomer and the CP43-less PSII monomer (a marker for ongoing PSII repair and reassembly). PSII repair and reassembly involve the breakage and formation of disulfide bonds among PSII proteins. Interestingly, the recombinant LQY1 protein demonstrates a protein disulfide isomerase activity. LQY1 is more abundant in stroma-exposed thylakoids, where key steps of PSII repair and reassembly take place. The absence of the LQY1 protein accelerates turnover and synthesis of PSII reaction center protein D1. These results suggest that the LQY1 protein may be involved in maintaining PSII activity under high light by regulating repair and reassembly of PSII complexes. PMID:21586683

  3. The Mcm2-7-interacting domain of human mini-chromosome maintenance 10 (Mcm10) protein is important for stable chromatin association and origin firing.

    PubMed

    Izumi, Masako; Mizuno, Takeshi; Yanagi, Ken-Ichiro; Sugimura, Kazuto; Okumura, Katsuzumi; Imamoto, Naoko; Abe, Tomoko; Hanaoka, Fumio

    2017-08-04

    The protein mini-chromosome maintenance 10 (Mcm10) was originally identified as an essential yeast protein in the maintenance of mini-chromosome plasmids. Subsequently, Mcm10 has been shown to be required for both initiation and elongation during chromosomal DNA replication. However, it is not fully understood how the multiple functions of Mcm10 are coordinated or how Mcm10 interacts with other factors at replication forks. Here, we identified and characterized the Mcm2-7-interacting domain in human Mcm10. The interaction with Mcm2-7 required the Mcm10 domain that contained amino acids 530-655, which overlapped with the domain required for the stable retention of Mcm10 on chromatin. Expression of truncated Mcm10 in HeLa cells depleted of endogenous Mcm10 via siRNA revealed that the Mcm10 conserved domain (amino acids 200-482) is essential for DNA replication, whereas both the conserved and the Mcm2-7-binding domains were required for its full activity. Mcm10 depletion reduced the initiation frequency of DNA replication and interfered with chromatin loading of replication protein A, DNA polymerase (Pol) α, and proliferating cell nuclear antigen, whereas the chromatin loading of Cdc45 and Pol ϵ was unaffected. These results suggest that human Mcm10 is bound to chromatin through the interaction with Mcm2-7 and is primarily involved in the initiation of DNA replication after loading of Cdc45 and Pol ϵ. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. A chloroplast-targeted DnaJ protein contributes to maintenance of photosystem II under chilling stress

    PubMed Central

    Meng, Qingwei

    2014-01-01

    DnaJ proteins act as essential molecular chaperones in protein homeostasis and protein complex stabilization under stress conditions. The roles of a tomato (Lycopersicon esculentum) chloroplast-targeted DnaJ protein (LeCDJ1), whose expression was upregulated by treatment at 4 and 42 °C, and with high light, NaCl, polyethylene glycol, and H2O2, were investigated here using sense and antisense transgenic tomatoes. The sense plants exhibited not only higher chlorophyll content, fresh weight and net photosynthetic rate, but also lower accumulation of reactive oxygen species and membrane damage under chilling stress. Moreover, the maximal photochemistry efficiency of photosystem II (PSII) (F v/F m) and D1 protein content were higher in the sense plants and lower in the antisense plants, and the photoinhibitory quenching was lower in the sense plants and higher in the antisense plants, suggesting that the inhibition of PSII was less severe in the sense plants and more severe in the antisense plants compared with the wild type. Furthermore, the PSII protein complexes were also more stable in the sense plants. Interestingly, the sense plants treated with streptomycin (SM), an inhibitor of organellar translation, still showed higher F v/F m, D1 protein content and PSII stability than the SM-untreated antisense plants. This finding suggested that the protective effect of LeCDJ1 on PSII was, at least partially, independent of D1 protein synthesis. Furthermore, chloroplast heat-shock protein 70 was identified as the partner of LeCDJ1. These results indicate that LeCDJ1 has essential functions in maintaining PSII under chilling stress. PMID:24227338

  5. Regulation of synaptic Rac1 activity, long-term potentiation maintenance, and learning and memory by BCR and ABR Rac GTPase-activating proteins.

    PubMed

    Oh, Daeyoung; Han, Seungnam; Seo, Jinsoo; Lee, Jae-Ran; Choi, Jeonghoon; Groffen, John; Kim, Karam; Cho, Yi Sul; Choi, Han-Saem; Shin, Hyewon; Woo, Jooyeon; Won, Hyejung; Park, Soon Kwon; Kim, Soo-Young; Jo, Jihoon; Whitcomb, Daniel J; Cho, Kwangwook; Kim, Hyun; Bae, Yong Chul; Heisterkamp, Nora; Choi, Se-Young; Kim, Eunjoon

    2010-10-20

    Rho family small GTPases are important regulators of neuronal development. Defective Rho regulation causes nervous system dysfunctions including mental retardation and Alzheimer's disease. Rac1, a member of the Rho family, regulates dendritic spines and excitatory synapses, but relatively little is known about how synaptic Rac1 is negatively regulated. Breakpoint cluster region (BCR) is a Rac GTPase-activating protein known to form a fusion protein with the c-Abl tyrosine kinase in Philadelphia chromosome-positive chronic myelogenous leukemia. Despite the fact that BCR mRNAs are abundantly expressed in the brain, the neural functions of BCR protein have remained obscure. We report here that BCR and its close relative active BCR-related (ABR) localize at excitatory synapses and directly interact with PSD-95, an abundant postsynaptic scaffolding protein. Mice deficient for BCR or ABR show enhanced basal Rac1 activity but only a small increase in spine density. Importantly, mice lacking BCR or ABR exhibit a marked decrease in the maintenance, but not induction, of long-term potentiation, and show impaired spatial and object recognition memory. These results suggest that BCR and ABR have novel roles in the regulation of synaptic Rac1 signaling, synaptic plasticity, and learning and memory, and that excessive Rac1 activity negatively affects synaptic and cognitive functions.

  6. Regulation of Synaptic Rac1 Activity, Long-Term Potentiation Maintenance, and Learning and Memory by BCR and ABR Rac GTPase-Activating Proteins

    PubMed Central

    Oh, Daeyoung; Han, Seungnam; Seo, Jinsoo; Lee, Jae-Ran; Choi, Jeonghoon; Groffen, John; Kim, Karam; Cho, Yi Sul; Choi, Han-Saem; Shin, Hyewon; Woo, Jooyeon; Won, Hyejung; Park, Soon Kwon; Kim, Soo-Young; Jo, Jihoon; Whitcomb, Daniel J.; Cho, Kwangwook; Kim, Hyun; Bae, Yong Chul; Heisterkamp, Nora; Choi, Se-Young; Kim, Eunjoon

    2016-01-01

    Rho family small GTPases are important regulators of neuronal development. Defective Rho regulation causes nervous system dysfunctions including mental retardation and Alzheimer’s disease. Rac1, a member of the Rho family, regulates dendritic spines and excitatory synapses, but relatively little is known about how synaptic Rac1 is negatively regulated. Breakpoint cluster region (BCR) is a Rac GTPase-activating protein known to form a fusion protein with the c-Abl tyrosine kinase in Philadelphia chromosome-positive chronic myelogenous leukemia. Despite the fact that BCR mRNAs are abundantly expressed in the brain, the neural functions of BCR protein have remained obscure. We report here that BCR and its close relative active BCR-related (ABR) localize at excitatory synapses and directly interact with PSD-95, an abundant postsynaptic scaffolding protein. Mice deficient for BCR or ABR show enhanced basal Rac1 activity but only a small increase in spine density. Importantly, mice lacking BCR or ABR exhibit a marked decrease in the maintenance, but not induction, of long-term potentiation, and show impaired spatial and object recognition memory. These results suggest that BCR and ABR have novel roles in the regulation of synaptic Rac1 signaling, synaptic plasticity, and learning and memory, and that excessive Rac1 activity negatively affects synaptic and cognitive functions. PMID:20962234

  7. Providing a diet containing only maintenance levels of energy and protein during the latter stages of pregnancy resulted in a prolonged delivery time during parturition in rats.

    PubMed

    Tanaka, Y; Kadokawa, H

    2012-01-01

    In mammals, a prolonged delivery time during parturition is dangerous for both mother and fetus, although the mechanisms that prolong delivery are unclear. To investigate whether nutrition affects delivery time, we administered two feeds containing maintenance (L-feed) or higher (H-feed) levels of energy and protein at different points during the latter half of pregnancy and compared the effects of the various treatments on delivery time in rats. After the rats had been maintained on the L-feed and then copulated on pro-oestrus (Day 0), pregnant females were randomly allocated to one of three groups: (1) the no-improvement group, which was fed L-feed throughout gestation; (2) the early group, which was fed L-feed until Day 11 of gestation and then switched to H-feed; and (3) the late group, which was fed L-feed until Day 16 of gestation and then switched to H-feed. There was no significant difference in the number of pups among the three groups. However, delivery time was significantly longer in the no-improvement group (73.7±5.2 min) than the early (46.9±5.6 min) and late (55.4±5.5 min) groups. Consuming a maintenance diet during the latter half of pregnancy resulted in a prolonged delivery time.

  8. The BEACH protein LRBA is required for hair bundle maintenance in cochlear hair cells and for hearing.

    PubMed

    Vogl, Christian; Butola, Tanvi; Haag, Natja; Hausrat, Torben J; Leitner, Michael G; Moutschen, Michel; Lefèbvre, Philippe P; Speckmann, Carsten; Garrett, Lillian; Becker, Lore; Fuchs, Helmut; Hrabe de Angelis, Martin; Nietzsche, Sandor; Kessels, Michael M; Oliver, Dominik; Kneussel, Matthias; Kilimann, Manfred W; Strenzke, Nicola

    2017-09-11

    Lipopolysaccharide-responsive beige-like anchor protein (LRBA) belongs to the enigmatic class of BEACH domain-containing proteins, which have been attributed various cellular functions, typically involving intracellular protein and membrane transport processes. Here, we show that LRBA deficiency in mice leads to progressive sensorineural hearing loss. In LRBA knockout mice, inner and outer hair cell stereociliary bundles initially develop normally, but then partially degenerate during the second postnatal week. LRBA deficiency is associated with a reduced abundance of radixin and Nherf2, two adaptor proteins, which are important for the mechanical stability of the basal taper region of stereocilia. Our data suggest that due to the loss of structural integrity of the central parts of the hair bundle, the hair cell receptor potential is reduced, resulting in a loss of cochlear sensitivity and functional loss of the fraction of spiral ganglion neurons with low spontaneous firing rates. Clinical data obtained from two human patients with protein-truncating nonsense or frameshift mutations suggest that LRBA deficiency may likewise cause syndromic sensorineural hearing impairment in humans, albeit less severe than in our mouse model. © 2017 The Authors.

  9. The conserved LIM domain-containing focal adhesion protein ZYX-1 regulates synapse maintenance in Caenorhabditis elegans

    PubMed Central

    Luo, Shuo; Schaefer, Anneliese M.; Dour, Scott; Nonet, Michael L.

    2014-01-01

    We describe the identification of zyxin as a regulator of synapse maintenance in mechanosensory neurons in C. elegans. zyx-1 mutants lacked PLM mechanosensory synapses as adult animals. However, most PLM synapses initially formed during development but were subsequently lost as the animals developed. Vertebrate zyxin regulates cytoskeletal responses to mechanical stress in culture. Our work provides in vivo evidence in support of such a role for zyxin. In particular, zyx-1 mutant synaptogenesis phenotypes were suppressed by disrupting locomotion of the mutant animals, suggesting that zyx-1 protects mechanosensory synapses from locomotion-induced forces. In cultured cells, zyxin is recruited to focal adhesions and stress fibers via C-terminal LIM domains and modulates cytoskeletal organization via the N-terminal domain. The synapse-stabilizing activity was mediated by a short isoform of ZYX-1 containing only the LIM domains. Consistent with this notion, PLM synaptogenesis was independent of α-actinin and ENA-VASP, both of which bind to the N-terminal domain of zyxin. Our results demonstrate that the LIM domain moiety of zyxin functions autonomously to mediate responses to mechanical stress and provide in vivo evidence for a role of zyxin in neuronal development. PMID:25252943

  10. The conserved LIM domain-containing focal adhesion protein ZYX-1 regulates synapse maintenance in Caenorhabditis elegans.

    PubMed

    Luo, Shuo; Schaefer, Anneliese M; Dour, Scott; Nonet, Michael L

    2014-10-01

    We describe the identification of zyxin as a regulator of synapse maintenance in mechanosensory neurons in C. elegans. zyx-1 mutants lacked PLM mechanosensory synapses as adult animals. However, most PLM synapses initially formed during development but were subsequently lost as the animals developed. Vertebrate zyxin regulates cytoskeletal responses to mechanical stress in culture. Our work provides in vivo evidence in support of such a role for zyxin. In particular, zyx-1 mutant synaptogenesis phenotypes were suppressed by disrupting locomotion of the mutant animals, suggesting that zyx-1 protects mechanosensory synapses from locomotion-induced forces. In cultured cells, zyxin is recruited to focal adhesions and stress fibers via C-terminal LIM domains and modulates cytoskeletal organization via the N-terminal domain. The synapse-stabilizing activity was mediated by a short isoform of ZYX-1 containing only the LIM domains. Consistent with this notion, PLM synaptogenesis was independent of α-actinin and ENA-VASP, both of which bind to the N-terminal domain of zyxin. Our results demonstrate that the LIM domain moiety of zyxin functions autonomously to mediate responses to mechanical stress and provide in vivo evidence for a role of zyxin in neuronal development.

  11. WHC Maintenance

    NASA Image and Video Library

    2009-04-28

    ISS019-E-011464 (28 April 2009) --- Japan Aerospace Exploration Agency (JAXA) astronaut Koichi Wakata, Expedition 19/20 flight engineer, performs in-flight maintenance on the waste and hygiene compartment located in the Destiny laboratory of the International Space Station.

  12. WHC Maintenance

    NASA Image and Video Library

    2009-04-28

    ISS019-E-011471 (28 April 2009) --- Japan Aerospace Exploration Agency (JAXA) astronaut Koichi Wakata, Expedition 19/20 flight engineer, performs in-flight maintenance on the waste and hygiene compartment located in the Destiny laboratory of the International Space Station.

  13. Mdm31 and Mdm32 are inner membrane proteins required for maintenance of mitochondrial shape and stability of mitochondrial DNA nucleoids in yeast.

    PubMed

    Dimmer, Kai Stefan; Jakobs, Stefan; Vogel, Frank; Altmann, Katrin; Westermann, Benedikt

    2005-01-03

    The MDM31 and MDM32 genes are required for normal distribution and morphology of mitochondria in the yeast Saccharomyces cerevisiae. They encode two related proteins located in distinct protein complexes in the mitochondrial inner membrane. Cells lacking Mdm31 and Mdm32 harbor giant spherical mitochondria with highly aberrant internal structure. Mitochondrial DNA (mtDNA) is instable in the mutants, mtDNA nucleoids are disorganized, and their association with Mmm1-containing complexes in the outer membrane is abolished. Mutant mitochondria are largely immotile, resulting in a mitochondrial inheritance defect. Deletion of either one of the MDM31 and MDM32 genes is synthetically lethal with deletion of either one of the MMM1, MMM2, MDM10, and MDM12 genes, which encode outer membrane proteins involved in mitochondrial morphogenesis and mtDNA inheritance. We propose that Mdm31 and Mdm32 cooperate with Mmm1, Mmm2, Mdm10, and Mdm12 in maintenance of mitochondrial morphology and mtDNA.

  14. Slow Co-Evolution of the MAGO and Y14 Protein Families Is Required for the Maintenance of Their Obligate Heterodimerization Mode

    PubMed Central

    He, Chaoying

    2014-01-01

    The exon junction complex (EJC) plays important roles in RNA metabolisms and the development of eukaryotic organisms. MAGO (short form of MAGO NASHI) and Y14 (also Tsunagi or RBM8) are the EJC core components. Their biological roles have been well investigated in various species, but the evolutionary patterns of the two gene families and their protein-protein interactions are poorly known. Genome-wide survey suggested that the MAGO and Y14 two gene families originated in eukaryotic organisms with the maintenance of a low copy. We found that the two protein families evolved slowly; however, the MAGO family under stringent purifying selection evolved more slowly than the Y14 family that was under relative relaxed purifying selection. MAGO and Y14 were obliged to form heterodimer in a eukaryotic organism, and this obligate mode was plesiomorphic. Lack of binding of MAGO to Y14 as functional barrier was observed only among distantly species, suggesting that a slow co-evolution of the two protein families. Inter-protein co-evolutionary signal was further quantified in analyses of the Tol-MirroTree and co-evolution analysis using protein sequences. About 20% of the 41 significantly correlated mutation groups (involving 97 residues) predicted between the two families was clade-specific. Moreover, around half of the predicted co-evolved groups and nearly all clade-specific residues fell into the minimal interaction domains of the two protein families. The mutagenesis effects of the clade-specific residues strengthened that the co-evolution is required for obligate MAGO-Y14 heterodimerization mode. In turn, the obliged heterodimerization in an organism serves as a strong functional constraint for the co-evolution of the MAGO and Y14 families. Such a co-evolution allows maintaining the interaction between the proteins through large evolutionary time scales. Our work shed a light on functional evolution of the EJC genes in eukaryotes, and facilitates to understand the co

  15. Determination of energy and protein requirement for maintenance and growth and evaluation for the effects of gender upon nutrient requirement in Dorper × Hu Crossbred Lambs.

    PubMed

    Nie, Hai Tao; Zhang, Hao; You, Ji Hao; Wang, Feng

    2015-06-01

    This study aimed to determine energy and protein requirement of Dorper × Hu crossbred lambs and further to evaluate the effect of gender upon nutrient requirement parameters. Forty-two female lambs (18.60 ± 1.57 kg) and 42 male lambs (18.30 ± 1.28 kg) were used. In comparative slaughter trial, 30 of animals from each gender group were randomly selected and assigned to ad libitum (AL), low restriction (LR) and high restriction (HR) group, and then were slaughtered when lambs under AL treatment reached target BW of 20, 28, and 35 kg, to determine body energy and nitrogen retained. In digestibility trial, remaining 12 female (18.01 ± 1.66 kg) and 12 male lambs (18.43 ± 1.17 kg) were randomly assigned to three feeding treatments in accordance with the design of comparative slaughter trial, to evaluate dietary energetic values at different feed intake levels. The combined data indicated that metabolizable energy (ME) requirement for maintenance (MEm; 400.61 ± 20.31 vs. 427.24 ± 18.70 kJ kg(-1) of shrunk BW(0.75); SBW(0.75)), partial efficiency of ME utilization for maintenance (k m; 0.64 ± 0.02 vs. 0.65 ± 0.03), partial efficiency of ME utilization for growth (k g ; 0.42 ± 0.03 vs. 0.44 ± 0.02), and net protein (NP) requirement for maintenance (NPm; 1.83 ± 0.17 vs. 1.99 ± 0.28 g kg(-1) of SBW(0.75)) did not differ (P > 0.05) due to gender; although not statistically different, the mean value of Net energy (NE) requirement for maintenance (NEm) for male lambs (260.62 ± 13.21 kJ kg(-1) of SBW(0.75)) were 5 % greater than that (274.16 ± 11.99 kJ kg(-1) of SBW(0.75)) of female lambs. Additionally, rams have greater amounts of NP requirement for growth (NPg, 15.94 to 44.32 g d(-1)) than those of ewes (13.07 to 32.95 g d(-1)) at the similar condition of BW and ADG. In conclusion, we suggested that our results of energy and protein requirement for growth ranged between the NRC

  16. Maintenance of the marginal-zone B cell compartment specifically requires the RNA-binding protein ZFP36L1.

    PubMed

    Newman, Rebecca; Ahlfors, Helena; Saveliev, Alexander; Galloway, Alison; Hodson, Daniel J; Williams, Robert; Besra, Gurdyal S; Cook, Charlotte N; Cunningham, Adam F; Bell, Sarah E; Turner, Martin

    2017-04-10

    RNA-binding proteins of the ZFP36 family are best known for inhibiting the expression of cytokines through binding to AU-rich elements in the 3' untranslated region and promoting mRNA decay. Here we identified an indispensable role for ZFP36L1 as the regulator of a post-transcriptional hub that determined the identity of marginal-zone B cells by promoting their proper localization and survival. ZFP36L1 controlled a gene-expression program related to signaling, cell adhesion and locomotion; it achieved this in part by limiting expression of the transcription factors KLF2 and IRF8, which are known to enforce the follicular B cell phenotype. These mechanisms emphasize the importance of integrating transcriptional and post-transcriptional processes by RNA-binding proteins for maintaining cellular identity among closely related cell types.

  17. Body protein index based on bioelectrical impedance analysis is a useful new marker assessing nutritional status: applications to patients with chronic renal failure on maintenance dialysis.

    PubMed

    Nakao, Toshiyuki; Kanazawa, Yoshie; Nagaoka, Yume; Iwasawa, Hideaki; Uchinaga, Asako; Matsumoto, Hiroshi; Okada, Tomonari; Yoshino, Maki

    2007-01-01

    Evaluation and monitoring of nutritional status is a fundamental concept in providing nutritional care to patients with end-stage renal failure. There have been, however, few practically available indices assessing whole body protein stores of patients. We enrolled 448 end-stage renal disease patients, 394 on maintenance hemodialysis (HD) and 54 on continuous ambulatory peritoneal dialysis (PD) in this study. 83 Age- and sex-matched subjects (controls) whose creatinine clearance was more than 70 ml/min and urinary protein excretion was less than 1.0 g/day were also recruited for comparison. To assess whole body somatic protein stores, we devised the body protein index (BPI). The volume of body protein mass was measured by multifrequency bioelectrical impedance analysis and then BPI was calculated as body protein mass (kg) divided by height in meters (m2). Based on BPI, we defined the nutritional status of the patients as normal if the value was within -10% of the mean value of control subjects, -10 to -14% as mild malnutrition, -15 to -19% as moderate malnutrition, and <-20% as severe malnutrition. The required time for measurement was 5.2 +/- 1.3 min and coefficient of variation of measurements was 0.8 +/- 0.2%. Among men the mean BPI in both HD and PD patients was significantly lower than those of control subjects (4.25 +/- 0.37, 4.38 +/- 0.34 vs. 4.72 +/- 0.37 kg/m2, p < 0.001). In women, BPI was significantly lower in HD patients than in control subjects (3.65 +/- 0.34 vs. 4.00 +/- 0.34 kg/m2, p < 0.033), whereas only a nonsignificant lower tendency was found in PD patients (3.83 +/- 0.39 kg/m2, p = 0.067). There were no significant differences in BPI values between diabetic and non-diabetic subjects, both in men (4.26 +/- 0.41 vs. 4.25 +/- 0.36 kg/m2) and women (3.69 +/- 0.36 vs. 3.65 +/- 0.34 kg/m2). Based on BPI nutritional categories, 113 (28.7%) of all HD patients were classified as having mild malnutrition, 57 (14.5%) as having moderate malnutrition, 40

  18. Phosphorylation state of a Tob/BTG protein, FOG-3, regulates initiation and maintenance of the Caenorhabditis elegans sperm fate program.

    PubMed

    Lee, Myon-Hee; Kim, Kyung Won; Morgan, Clinton T; Morgan, Dyan E; Kimble, Judith

    2011-05-31

    FOG-3, the single Caenorhabditis elegans Tob/BTG protein, directs germ cells to adopt the sperm fate at the expense of oogenesis. Importantly, FOG-3 activity must be maintained for the continued production of sperm that is typical of the male sex. Vertebrate Tob proteins have antiproliferative activity and ERK phosphorylation of Tob proteins has been proposed to abrogate "antiproliferative" activity. Here we investigate FOG-3 phosphorylation and its effect on sperm fate specification. We found both phosphorylated and unphosphorylated forms of FOG-3 in nematodes. We then interrogated the role of FOG-3 phosphorylation in sperm fate specification. Specifically, we assayed FOG-3 transgenes for rescue of a fog-3 null mutant. Wild-type FOG-3 rescued both initiation and maintenance of sperm fate specification. A FOG-3 mutant with its four consensus ERK phosphorylation sites substituted to alanines, called FOG-3(4A), rescued partially: sperm were made transiently but not continuously in both sexes. A different FOG-3 mutant with its sites substituted to glutamates, called FOG-3(4E), had no rescuing activity on its own, but together with FOG-3(4A) rescue was complete. Thus, when FOG-3(4A) and FOG-3(4E) were both introduced into the same animals, sperm fate specification was not only initiated but also maintained, resulting in continuous spermatogenesis in males. Our findings suggest that unphosphorylated FOG-3 initiates the sperm fate program and that phosphorylated FOG-3 maintains that program for continued sperm production typical of males. We discuss implications of our results for Tob/BTG proteins in vertebrates.

  19. Directed Molecular Evolution of an Engineered Gammaretroviral Envelope Protein with Dual Receptor Use Shows Stable Maintenance of Both Receptor Specificities

    PubMed Central

    Friis, Kristina Pagh; Iturrioz, Xavier; Thomsen, Jonas; Alvear-Perez, Rodrigo; Bahrami, Shervin; Llorens-Cortes, Catherine

    2015-01-01

    ABSTRACT We have previously reported the construction of a murine leukemia virus-based replication-competent gammaretrovirus (SL3-AP) capable of utilizing the human G protein-coupled receptor APJ (hAPJ) as its entry receptor and its natural receptor, the murine Xpr1 receptor, with equal affinities. The apelin receptor has previously been shown to function as a coreceptor for HIV-1, and thus, adaptation of the viral vector to this receptor is of significant interest. Here, we report the molecular evolution of the SL3-AP envelope protein when the virus is cultured in cells harboring either the Xpr1 or the hAPJ receptor. Interestingly, the dual receptor affinity is maintained even after 10 passages in these cells. At the same time, the chimeric viral envelope protein evolves in a distinct pattern in the apelin cassette when passaged on D17 cells expressing hAPJ in three separate molecular evolution studies. This pattern reflects selection for reduced ligand-receptor interaction and is compatible with a model in which SL3-AP has evolved not to activate hAPJ receptor internalization. IMPORTANCE Few successful examples of engineered retargeting of a retroviral vector exist. The engineered SL3-AP envelope is capable of utilizing either the murine Xpr1 or the human APJ receptor for entry. In addition, SL3-AP is the first example of an engineered retrovirus retaining its dual tropism after several rounds of passaging on cells expressing only one of its receptors. We demonstrate that the virus evolves toward reduced ligand-receptor affinity, which sheds new light on virus adaptation. We provide indirect evidence that such reduced affinity leads to reduced receptor internalization and propose a novel model in which too rapid receptor internalization may decrease virus entry. PMID:26608314

  20. Membrane frizzled-related protein is necessary for the normal development and maintenance of photoreceptor outer segments.

    PubMed

    Won, Jungyeon; Smith, Richard S; Peachey, Neal S; Wu, Jiang; Hicks, Wanda L; Naggert, Jürgen K; Nishina, Patsy M

    2008-01-01

    A 4 base pair deletion in a splice donor site of the Mfrp (membrane-type frizzled-related protein) gene, herein referred to as Mfrprd6/rd6, is predicted to lead to the skipping of exon 4 and photoreceptor degeneration in retinal degeneration 6 (rd6) mutant mice. Little, however, is known about the function of the protein or how the mutation causes the degenerative retinal phenotype. Here we examine ultrastructural changes in the retina of Mfrprd6/rd6 mice to determine the earliest effects of the mutation. We also extend the reported observations of the expression pattern of the dicistronic Mfrp/C1qtnf5 message and the localization of these and other retinal pigment epithelium (RPE) and retinal proteins during development and assess the ability of RPE cells to phagocytize outer segments (OSs) in mutant and wild-type (WT) mice. At the ultrastructural level, OSs do not develop normally in Mfrprd6/rd6 mutants. They are disorganized and become progressively shorter as mutant mice age. Additionally, there are focal areas in which there is a reduction of apical RPE microvilli. At P25, the rod electroretinogram (ERG) a-wave of Mfrprd6/rd6 mice is reduced in amplitude by ~50% as are ERG components generated by the RPE. Examination of beta-catenin localization and Fos and Tcf-1 expression, intermediates of the canonical Wnt pathway, showed that they were not different between mutant and WT mice, suggesting that MFRP may operate through an alternative pathway. Finally, impaired OS phagocytosis was observed in Mfrprd6/rd6 mice both in standard ambient lighting conditions and with bright light exposure when compared to WT controls.

  1. Periodontal maintenance.

    PubMed

    Tan, A E S

    2009-09-01

    The main goal of periodontal therapy is to establish an oral environment compatible with periodontal health by the physical disruption of the plaque biofilm and adjunctive chemical means if required. Implicit in this objective is the ongoing requirement of detection and interception of new and recurrent disease, which continues at selected intervals for the life of the dentition after the initial ("active") phase of periodontal treatment. This concept of ongoing periodontal maintenance therapy has been embraced as the mandatory requirement for favourable periodontal outcomes based on institutional clinical trials and in practice-based studies in various parts of the world. This review examines the ramifications of periodontal maintenance therapy based upon a multi-level assessment of logistic issues and risk factors at three levels: (1) The patient level - treatment time; patient attendance compliance; and homecare measures, antiseptics/antibiotics and smoking. (2) The level of the individual tooth - tooth loss; and evaluation of success versus survival. (3) The level of each tooth surface ("site") - probing depth, loss of attachment and bleeding on probing; and changes in clinical attachment levels. In spite of the diversity of studies conducted, there is agreement on the efficacy of periodontal maintenance therapy when compared with studies on untreated populations and in treated cases that were not maintained.

  2. MICROTUBULE-ASSOCIATED PROTEIN65 is essential for maintenance of phragmoplast bipolarity and formation of the cell plate in Physcomitrella patens.

    PubMed

    Kosetsu, Ken; de Keijzer, Jeroen; Janson, Marcel E; Goshima, Gohta

    2013-11-01

    The phragmoplast, a plant-specific apparatus that mediates cytokinesis, mainly consists of microtubules (MTs) arranged in a bipolar fashion, such that their plus ends interdigitate at the equator. Membrane vesicles are thought to move along the MTs toward the equator and fuse to form the cell plate. Although several genes required for phragmoplast MT organization have been identified, the mechanisms that maintain the bipolarity of phragmoplasts remain poorly understood. Here, we show that engaging phragmoplast MTs in a bipolar fashion in protonemal cells of the moss Physcomitrella patens requires the conserved MT cross-linking protein MICROTUBULE-ASSOCIATED PROTEIN65 (MAP65). Simultaneous knockdown of the three MAP65s expressed in those cells severely compromised MT interdigitation at the phragmoplast equator after anaphase onset, resulting in the collapse of the phragmoplast in telophase. Cytokinetic vesicles initially localized to the anaphase midzone as normal but failed to further accumulate in the next several minutes, although the bipolarity of the MT array was preserved. Our data indicate that the presence of bipolar MT arrays is insufficient for vesicle accumulation at the equator and further suggest that MAP65-mediated MT interdigitation is a prerequisite for maintenance of bipolarity of the phragmoplast and accumulation and/or fusion of cell plate-destined vesicles at the equatorial plane.

  3. Mitochondrial p32 Protein Is a Critical Regulator of Tumor Metabolism via Maintenance of Oxidative Phosphorylation ▿

    PubMed Central

    Fogal, Valentina; Richardson, Adam D.; Karmali, Priya P.; Scheffler, Immo E.; Smith, Jeffrey W.; Ruoslahti, Erkki

    2010-01-01

    p32/gC1qR/C1QBP/HABP1 is a mitochondrial/cell surface protein overexpressed in certain cancer cells. Here we show that knocking down p32 expression in human cancer cells strongly shifts their metabolism from oxidative phosphorylation (OXPHOS) to glycolysis. The p32 knockdown cells exhibited reduced synthesis of the mitochondrial-DNA-encoded OXPHOS polypeptides and were less tumorigenic in vivo. Expression of exogenous p32 in the knockdown cells restored the wild-type cellular phenotype and tumorigenicity. Increased glucose consumption and lactate production, known as the Warburg effect, are almost universal hallmarks of solid tumors and are thought to favor tumor growth. However, here we show that a protein regularly overexpressed in some cancers is capable of promoting OXPHOS. Our results indicate that high levels of glycolysis, in the absence of adequate OXPHOS, may not be as beneficial for tumor growth as generally thought and suggest that tumor cells use p32 to regulate the balance between OXPHOS and glycolysis. PMID:20100866

  4. Intradialytic parenteral nutrition in maintenance hemodialysis patients suffering from protein-energy wasting. Results of a multicenter, open, prospective, randomized trial.

    PubMed

    Marsen, Tobias A; Beer, Justinus; Mann, Helmut

    2017-02-01

    Protein-energy wasting (PEW) is increasingly becoming a clinical problem in maintenance hemodialysis patients and guidelines call for nutritional interventions. Serum prealbumin (transthyretin) represents a critical nutritional marker positively correlated with patient survival and negatively correlated with morbidity. Nutritional counseling, oral supplementation as well as intradialytic parenteral nutrition (IDPN) are recommended to fight PEW, however clinical trials on their use are scarce. We conducted a prospective, multicenter, randomized, open-label, controlled, parallel-group Phase IV clinical trial in 107 maintenance hemodialysis patients suffering from PEW to assess the impact of IDPN on prealbumin and other biochemical and clinical parameters reflecting nutritional status. Patients randomized to the intervention group received standardized nutritional counseling plus IDPN three times weekly over 16 weeks followed by a treatment-free period of 12 weeks. The control group received standardized nutritional counseling only. Main trial inclusion criteria included moderate to severe malnutrition (SGA score B or C), maintenance hemodialysis therapy (3 times per week) for more than six months, and presence of two out of the following three criteria: albumin <35 g/L, prealbumin <250 mg/L, phase angle alpha <4.5° assessed by bioelectrical impedance analysis (BIA). Changes in serum prealbumin, albumin, transferrin, phase angle alpha, subjective global assessment (SGA) score and health-related quality of life using the 12-item short form health survey (SF-12) were investigated. IDPN significantly increased prealbumin (p < 0.05), showing rapid rise within 16 weeks of treatment and sustained response thereafter. In the full analysis set (n = 83), 41.0% of 39 patients receiving IDPN achieved a relevant (i.e., at least ≥15%) increase in prealbumin over baseline at week 4 compared to 20.5% of 44 patients in the control group. Considerably more patients with IDPN

  5. Comparison of C-reactive protein levels with delivered dose of Kt/V in patients with end-stage renal disease on maintenance hemodialysis.

    PubMed

    Rashid, Humayun

    2015-01-01

    The objective of our study is to compare the C-reactive protein (CRP) levels with the delivered dose of dialysis in terms of Kt/V in patients undergoing maintenance hemodialysis (HD). This is a comparative, cross-sectional survey. The study was conducted at the HD unit of the Shaikh Zayed Hospital, Lahore, Pakistan. Patients who fulfilled the inclusion and exclusion criteria were enrolled in this study. The delivered dose of HD (Kt/V) was assessed by an online clearance module (OCM) in Fresenius machines at the end of every dialysis session and the weekly Kt/V was determined by adding all three Kt/V values. The serum CRP sample was taken after each session of HD and the mean CRP was calculated and considered elevated if it was >6 mg/dL. Both weekly Kt/V and CRP values were entered in a pre-designed proforma. Data were analyzed by using statistical software SPSS and P-value £0.05 was considered significant. Of 100 patients on maintenance HD, high serum CRP level (>6 mg/dL) was found in 38 patients. When the Kt/V was compared with the CRP level, there was a negative correlation between the two parameters (r = 0.212, P = 0.032). Low Kt/V means dialysis inadequacy, which is associated with chronic inflammatory state, resulting in high CRP levels. We suggest that the quality of life of dialysis patients can be improved by offering an adequate dose of HD reflected by Kt/V ≥3.6/week.

  6. Assessing the Association between Serum Ferritin, Transferrin Saturation, and C-Reactive Protein in Northern Territory Indigenous Australian Patients with High Serum Ferritin on Maintenance Haemodialysis

    PubMed Central

    Lawton, Paul D.; Barzi, Federica; Cass, Alan; Hughes, Jaquelyne T.

    2017-01-01

    Objective. To determine the significance of high serum ferritin observed in Indigenous Australian patients on maintenance haemodialysis in the Northern Territory, we assessed the relationship between ferritin and transferrin saturation (TSAT) as measures of iron status and ferritin and C-reactive protein (CRP) as markers of inflammation. Methods. We performed a retrospective cohort analysis of data from adult patients (≥18 years) on maintenance haemodialysis (>3 months) from 2004 to 2011. Results. There were 1568 patients. The mean age was 53.9 (11.9) years. 1244 (79.3%) were Indigenous. 44.2% (n = 693) were male. Indigenous patients were younger (mean age [52.3 (11.1) versus 57.4 (15.2), p < 0.001]) and had higher CRP [14.7 mg/l (7–35) versus 5.9 mg/l (1.9–17.5), p < 0.001], higher median serum ferritin [1069 µg/l (668–1522) versus 794.9 µg/l (558.5–1252.0), p < 0.001], but similar transferrin saturation [26% (19–37) versus 28% (20–38), p = 0.516]. We observed a small positive correlation between ferritin and TSAT (r2 = 0.11, p < 0.001), no correlation between ferritin and CRP (r2 = 0.001, p < 0.001), and positive association between high serum ferritin and TSAT (p < 0.001), Indigenous ethnicity (p < 0.001), urea reduction ratio (p = 0.001), and gender (p < 0.001) after adjustment in mixed regression analysis. Conclusion. Serum ferritin and TSAT may inadequately reflect iron status in this population. The high ferritin was poorly explained by inflammation. PMID:28243472

  7. Early and Late Loss of the Cytoskeletal Scaffolding Protein, Ankyrin G Reveals its Role in Maturation and Maintenance of Nodes of Ranvier in Myelinated Axons.

    PubMed

    Saifetiarova, Julia; Taylor, Anna M; Bhat, Manzoor A

    2017-02-01

    The mechanisms that govern node of Ranvier organization, stability and long-term maintenance remain to be fully elucidated. One of the molecular components of the node is the cytoskeletal scaffolding protein, Ankyrin G (AnkG), which interacts with multiple members of the nodal complex. The role of AnkG in nodal organization and maintenance is still not clearly defined, as to whether AnkG functions as an initial nodal organizer or whether it functions as a nodal stabilizer after the nodal complex has been assembled. Using a mouse model system, we report here that perinatal and juvenile neuronal ablation of AnkG has differential consequences on nodal stability. Early loss of AnkG creates immature nodes with abnormal morphology, which undergo accelerated destabilization within a month, resulting in rapid NaV channel and βIV Spectrin loss with reduced effects on Neurofascin 186. On the other hand, late ablation of AnkG from established nodal complexes leads to slow but progressive nodal destabilization over 10 months, primarily affecting βIV Spectrin, followed by NaV channels, with modest impact on Neurofascin 186. We also show that Ankyrin R and βI Spectrin are not sufficient to prevent nodal disorganization after AnkG ablation. Additionally, nodal disorganization in both early and late AnkG mutants is accompanied by axonal pathology and neurological dysfunction. Together, our results suggest that AnkG plays an indispensable role in maturation and long-term stabilization of the newly assembled nodal complex, and that loss of AnkG after nodal stabilization does not lead to rapid nodal disassembly but loss of specific nodal components in a time-dependent manner.

  8. The human Cranio Facial Development Protein 1 (Cfdp1) gene encodes a protein required for the maintenance of higher-order chromatin organization

    PubMed Central

    Messina, Giovanni; Atterrato, Maria Teresa; Prozzillo, Yuri; Piacentini, Lucia; Losada, Ana; Dimitri, Patrizio

    2017-01-01

    The human Cranio Facial Development Protein 1 (Cfdp1) gene maps to chromosome 16q22.2-q22.3 and encodes the CFDP1 protein, which belongs to the evolutionarily conserved Bucentaur (BCNT) family. Craniofacial malformations are developmental disorders of particular biomedical and clinical interest, because they represent the main cause of infant mortality and disability in humans, thus it is important to understand the cellular functions and mechanism of action of the CFDP1 protein. We have carried out a multi-disciplinary study, combining cell biology, reverse genetics and biochemistry, to provide the first in vivo characterization of CFDP1 protein functions in human cells. We show that CFDP1 binds to chromatin and interacts with subunits of the SRCAP chromatin remodeling complex. An RNAi-mediated depletion of CFDP1 in HeLa cells affects chromosome organization, SMC2 condensin recruitment and cell cycle progression. Our findings provide new insight into the chromatin functions and mechanisms of the CFDP1 protein and contribute to our understanding of the link between epigenetic regulation and the onset of human complex developmental disorders. PMID:28367969

  9. Alzheimer's disease therapeutics targeted to the control of amyloid precursor protein translation: maintenance of brain iron homeostasis.

    PubMed

    Bandyopadhyay, Sanghamitra; Rogers, Jack T

    2014-04-15

    The neurotoxicity of amyloid beta (Aβ), a major cleavage product of the amyloid precursor protein (APP), is enhanced by iron, as found in the amyloid plaques of Alzheimer's disease (AD) patients. By contrast, the long-known neuroprotective activity of APP is evident after α-secretase cleavage of the precursor to release sAPPα, and depends on the iron export actions of APP itself. The latter underlie its neurotrophic and protective effects in facilitating the homeostatic actions of ferroportin mediated-iron export. Thus APP-dependent iron export may alleviate oxidative stress by minimizing labile iron thus protecting neurons from iron overload during stroke and hemorrhage. Consistent with this, altered phosphorylation of iron-regulatory protein-1 (IRP1) and its signaling processes play a critical role in modulating APP translation via the 5' untranslated region (5'UTR) of its transcript. The APP 5'UTR region encodes a functional iron-responsive element (IRE) RNA stem loop that represents a potential target for modulating APP production. Targeted regulation of APP gene expression via the modulation of 5'UTR sequence function represents a novel approach for the potential treatment of AD since altering APP translation can be used to improve both the protective brain iron balance and provide anti-amyloid efficacy. Approved drugs including paroxetine and desferrioxamine and several novel compounds have been identified that suppress abnormal metal-promoted Aβ accumulation with a subset of these acting via APP 5'UTR-dependent mechanisms to modulate APP translation and cleavage to generate the non-toxic sAPPα.

  10. The Protein Kinase Cδ Catalytic Fragment Is Critical for Maintenance of the G2/M DNA Damage Checkpoint*

    PubMed Central

    LaGory, Edward L.; Sitailo, Leonid A.; Denning, Mitchell F.

    2010-01-01

    Protein kinase Cδ (PKCδ) is an essential component of the intrinsic apoptotic program. Following DNA damage, such as exposure to UV radiation, PKCδ is cleaved in a caspase-dependent manner, generating a constitutively active catalytic fragment (PKCδ-cat), which is necessary and sufficient for keratinocyte apoptosis. We found that in addition to inducing apoptosis, expression of PKCδ-cat caused a pronounced G2/M cell cycle arrest in both primary human keratinocytes and immortalized HaCaT cells. Consistent with a G2/M arrest, PKCδ-cat induced phosphorylation of Cdk1 (Tyr15), a critical event in the G2/M checkpoint. Treatment with the ATM/ATR inhibitor caffeine was unable to prevent PKCδ-cat-induced G2/M arrest, suggesting that PKCδ-cat is functioning downstream of ATM/ATR in the G2/M checkpoint. To better understand the role of PKCδ and PKCδ-cat in the cell cycle response to DNA damage, we exposed wild-type and PKCδ null mouse embryonic fibroblasts (MEFs) to UV radiation. Wild-type MEFs underwent a pronounced G2/M arrest, Cdk1 phosphorylation, and induction of apoptosis following UV exposure, whereas PKCδ null MEFs were resistant to these effects. Expression of PKCδ-green fluorescent protein, but not caspase-resistant or kinase-inactive PKCδ, was able to restore G2/M checkpoint integrity in PKCδ null MEFs. The function of PKCδ in the DNA damage-induced G2/M cell cycle checkpoint may be a critical component of its tumor suppressor function. PMID:19917613

  11. Protein-Pacing Caloric-Restriction Enhances Body Composition Similarly in Obese Men and Women during Weight Loss and Sustains Efficacy during Long-Term Weight Maintenance

    PubMed Central

    Arciero, Paul J.; Edmonds, Rohan; He, Feng; Ward, Emery; Gumpricht, Eric; Mohr, Alex; Ormsbee, Michael J.; Astrup, Arne

    2016-01-01

    Short-Term protein-pacing (P; ~6 meals/day, >30% protein/day) and caloric restriction (CR, ~25% energy deficit) improves total (TBF), abdominal (ABF) and visceral (VAT) fat loss, energy expenditure, and biomarkers compared to heart healthy (HH) recommendations (3 meals/day, 15% protein/day) in obese adults. Less is known whether obese men and women respond similarly to P-CR during weight loss (WL) and whether a modified P-CR (mP-CR) is more efficacious than a HH diet during long-term (52 week) weight maintenance (WM). The purposes of this study were to evaluate the efficacy of: (1) P-CR on TBF, ABF, resting metabolic rate (RMR), and biomarkers between obese men and women during WL (weeks 0–12); and (2) mP-CR compared to a HH diet during WM (weeks 13–64). During WL, men (n = 21) and women (n = 19) were assessed for TBF, ABF, VAT, RMR, and biomarkers at weeks 0 (pre) and 12 (post). Men and women had similar reductions (p < 0.01) in weight (10%), TBF (19%), ABF (25%), VAT (33%), glucose (7%–12%), insulin (40%), leptin (>50%) and increase in % lean body mass (9%). RMR (kcals/kg bodyweight) was unchanged and respiratory quotient decreased 9%. Twenty-four subjects (mP-CR, n = 10; HH, n = 14) completed WM. mP-CR regained significantly less body weight (6%), TBF (12%), and ABF (17%) compared to HH (p < 0.05). Our results demonstrate P-CR enhances weight loss, body composition and biomarkers, and maintains these changes for 52-weeks compared to a traditional HH diet. PMID:27483317

  12. Protein-Pacing Caloric-Restriction Enhances Body Composition Similarly in Obese Men and Women during Weight Loss and Sustains Efficacy during Long-Term Weight Maintenance.

    PubMed

    Arciero, Paul J; Edmonds, Rohan; He, Feng; Ward, Emery; Gumpricht, Eric; Mohr, Alex; Ormsbee, Michael J; Astrup, Arne

    2016-07-30

    Short-Term protein-pacing (P; ~6 meals/day, >30% protein/day) and caloric restriction (CR, ~25% energy deficit) improves total (TBF), abdominal (ABF) and visceral (VAT) fat loss, energy expenditure, and biomarkers compared to heart healthy (HH) recommendations (3 meals/day, 15% protein/day) in obese adults. Less is known whether obese men and women respond similarly to P-CR during weight loss (WL) and whether a modified P-CR (mP-CR) is more efficacious than a HH diet during long-term (52 week) weight maintenance (WM). The purposes of this study were to evaluate the efficacy of: (1) P-CR on TBF, ABF, resting metabolic rate (RMR), and biomarkers between obese men and women during WL (weeks 0-12); and (2) mP-CR compared to a HH diet during WM (weeks 13-64). During WL, men (n = 21) and women (n = 19) were assessed for TBF, ABF, VAT, RMR, and biomarkers at weeks 0 (pre) and 12 (post). Men and women had similar reductions (p < 0.01) in weight (10%), TBF (19%), ABF (25%), VAT (33%), glucose (7%-12%), insulin (40%), leptin (>50%) and increase in % lean body mass (9%). RMR (kcals/kg bodyweight) was unchanged and respiratory quotient decreased 9%. Twenty-four subjects (mP-CR, n = 10; HH, n = 14) completed WM. mP-CR regained significantly less body weight (6%), TBF (12%), and ABF (17%) compared to HH (p < 0.05). Our results demonstrate P-CR enhances weight loss, body composition and biomarkers, and maintains these changes for 52-weeks compared to a traditional HH diet.

  13. Functional conservation of the pre-sensor one beta-finger hairpin (PS1-hp) structures in mini-chromosome maintenance proteins of Saccharomyces cerevisiae and archaea.

    PubMed

    Ramey, Christopher J; Sclafani, Robert A

    2014-05-28

    Mini-chromosome maintenance (MCM) proteins form complexes that are required for DNA replication and are highly conserved throughout evolution. The replicative helicase of eukaryotic organisms is composed of the six paralogs MCM2-7, which form a heterohexameric ring structure. In contrast, the structure of the archaean replicative MCM helicase is a single Mcm protein that forms a homohexameric complex. Atomic structures of archaeal MCMs have identified multiple beta-finger structures in Mcm proteins whose in vivo function is unknown. In the present study, we have investigated the physiological role of the pre-sensor 1 beta-hairpin (PS1-hp) beta-fingers of Saccharomyces cerevisiae Mcm4p and Mcm5p in DNA replication initiation and elongation in vivo. The PS1-hp beta-finger mutant of Mcm5p (mcm5-HAT K506A::URA3) has a growth defect at both 18° and 37°. Mutation of the Mcm4p PS1-hp beta-finger (mcm4-HA K658A::TRP1) does not have a growth defect, indicating different functional contributions of the PS1-hp beta-finger structures of different MCM helicase subunits. Both Mcm4p and Mcm5p PS1-hp beta-finger mutants can coimmunoprecipitate Mcm2p, indicating the formation of the hexameric MCM helicase complex. Both PS1-hp beta-finger mutants have a plasmid loss phenotype that is suppressible by origin dosage, indicating a defective replication initiation. Surprisingly, a defect in the binding of PS1-hp MCM mutants to origins of DNA replication was not found by chromatin immunoprecipitation, suggesting a novel interpretation in which the defect is in a subsequent step of DNA strand separation by the MCM helicase. The double mutant mcm4-HA K658A::TRP1 mcm5-HAT K506A::URA3 is lethal, displaying a terminal MCM mutant phenotype of large budded cells.

  14. Chromatin Modulatory Proteins and Olfactory Receptor Signaling in the Refinement and Maintenance of Fruitless Expression in Olfactory Receptor Neurons

    PubMed Central

    Li, Qingyun; Okuwa, Sumie; Peng, Bo; Wu, Jianni; Volkan, Pelin Cayirlioglu

    2016-01-01

    During development, sensory neurons must choose identities that allow them to detect specific signals and connect with appropriate target neurons. Ultimately, these sensory neurons will successfully integrate into appropriate neural circuits to generate defined motor outputs, or behavior. This integration requires a developmental coordination between the identity of the neuron and the identity of the circuit. The mechanisms that underlie this coordination are currently unknown. Here, we describe two modes of regulation that coordinate the sensory identities of Drosophila melanogaster olfactory receptor neurons (ORNs) involved in sex-specific behaviors with the sex-specific behavioral circuit identity marker fruitless (fru). The first mode involves a developmental program that coordinately restricts to appropriate ORNs the expression of fru and two olfactory receptors (Or47b and Ir84a) involved in sex-specific behaviors. This regulation requires the chromatin modulatory protein Alhambra (Alh). The second mode relies on the signaling from the olfactory receptors through CamK and histone acetyl transferase p300/CBP to maintain ORN-specific fru expression. Our results highlight two feed-forward regulatory mechanisms with both developmentally hardwired and olfactory receptor activity-dependent components that establish and maintain fru expression in ORNs. Such a dual mechanism of fru regulation in ORNs might be a trait of neurons driving plastic aspects of sex-specific behaviors. PMID:27093619

  15. Synapse formation and maintenance by C1q family proteins: a new class of secreted synapse organizers.

    PubMed

    Yuzaki, Michisuke

    2010-07-01

    Several C1q family members, especially the Cbln and C1q-like subfamilies, are highly and predominantly expressed in the central nervous system. Cbln1, a member of the Cbln subfamily, plays two unique roles at parallel fiber (PF)-Purkinje cell synapses in the cerebellum: the formation and stabilization of synaptic contact, and the control of functional synaptic plasticity by regulating the postsynaptic endocytotic pathway. The delta2 glutamate receptor (GluD2), which is predominantly expressed in Purkinje cells, plays similar critical roles in the cerebellum. In addition, viral expression of GluD2 or the application of recombinant Cbln1 induces PF-Purkinje cell synaptogenesis in vitro and in vivo. Antigen-unmasking methods were necessary to reveal the immunoreactivities for endogenous Cbln1 and GluD2 at the synaptic junction of PF synapses. We propose that Cbln1 and GluD2 are located at the synaptic cleft, where various proteins undergo intricate molecular interactions with each other, and serve as a bidirectional synaptic organizer.

  16. Interaction between basic residues of Epstein-Barr virus EBNA1 protein and cellular chromatin mediates viral plasmid maintenance.

    PubMed

    Kanda, Teru; Horikoshi, Naoki; Murata, Takayuki; Kawashima, Daisuke; Sugimoto, Atsuko; Narita, Yohei; Kurumizaka, Hitoshi; Tsurumi, Tatsuya

    2013-08-16

    The Epstein-Barr virus (EBV) genome is episomally maintained in latently infected cells. The viral protein EBNA1 is a bridging molecule that tethers EBV episomes to host mitotic chromosomes as well as to interphase chromatin. EBNA1 localizes to cellular chromosomes (chromatin) via its chromosome binding domains (CBDs), which are rich in glycine and arginine residues. However, the molecular mechanism by which the CBDs of EBNA1 attach to cellular chromatin is still under debate. Mutation analyses revealed that stepwise substitution of arginine residues within the CBD1 (amino acids 40-54) and CBD2 (amino acids 328-377) regions with alanines progressively impaired chromosome binding activity of EBNA1. The complete arginine-to-alanine substitutions within the CBD1 and -2 regions abolished the ability of EBNA1 to stably maintain EBV-derived oriP plasmids in dividing cells. Importantly, replacing the same arginines with lysines had minimal effect, if any, on chromosome binding of EBNA1 as well as on its ability to stably maintain oriP plasmids. Furthermore, a glycine-arginine-rich peptide derived from the CBD1 region bound to reconstituted nucleosome core particles in vitro, as did a glycine-lysine rich peptide, whereas a glycine-alanine rich peptide did not. These results support the idea that the chromosome binding of EBNA1 is mediated by electrostatic interactions between the basic amino acids within the CBDs and negatively charged cellular chromatin.

  17. Interaction between Basic Residues of Epstein-Barr Virus EBNA1 Protein and Cellular Chromatin Mediates Viral Plasmid Maintenance*

    PubMed Central

    Kanda, Teru; Horikoshi, Naoki; Murata, Takayuki; Kawashima, Daisuke; Sugimoto, Atsuko; Narita, Yohei; Kurumizaka, Hitoshi; Tsurumi, Tatsuya

    2013-01-01

    The Epstein-Barr virus (EBV) genome is episomally maintained in latently infected cells. The viral protein EBNA1 is a bridging molecule that tethers EBV episomes to host mitotic chromosomes as well as to interphase chromatin. EBNA1 localizes to cellular chromosomes (chromatin) via its chromosome binding domains (CBDs), which are rich in glycine and arginine residues. However, the molecular mechanism by which the CBDs of EBNA1 attach to cellular chromatin is still under debate. Mutation analyses revealed that stepwise substitution of arginine residues within the CBD1 (amino acids 40–54) and CBD2 (amino acids 328–377) regions with alanines progressively impaired chromosome binding activity of EBNA1. The complete arginine-to-alanine substitutions within the CBD1 and -2 regions abolished the ability of EBNA1 to stably maintain EBV-derived oriP plasmids in dividing cells. Importantly, replacing the same arginines with lysines had minimal effect, if any, on chromosome binding of EBNA1 as well as on its ability to stably maintain oriP plasmids. Furthermore, a glycine-arginine-rich peptide derived from the CBD1 region bound to reconstituted nucleosome core particles in vitro, as did a glycine-lysine rich peptide, whereas a glycine-alanine rich peptide did not. These results support the idea that the chromosome binding of EBNA1 is mediated by electrostatic interactions between the basic amino acids within the CBDs and negatively charged cellular chromatin. PMID:23836915

  18. C-reactive protein levels in patients on maintenance hemodialysis: reliability and reflection on the utility of single measurements.

    PubMed

    Stigant, Caroline E; Djurdjev, Ognjenka; Levin, Adeera

    2005-01-01

    Single C-reactive protein (CRP) values have been associated with death and cardiovascular disease in dialysis patients. We prospectively obtained multiple CRP values in stable patients, hypothesizing that values would remain stable in the absence of disease and that a single CRP value would be a reliable marker of risk. Four CRP values per week for three consecutive weeks were obtained in 10 clinically stable patients receiving conventional HD. Using prespecified cutoffs of 2.2 and 4.4 mg/l, the frequency of risk misclassification relative to the lowest CRP value obtained was determined. Within and between patient variability was also calculated. The median age was 54 years, and the average duration of dialysis was 41 months. Nine out of ten patients had at least one abnormal CRP value (>2.2 mg/l), six had all values elevated, and seven had an abnormal median CRP. The overall coefficient of reliability was 0.63 (95% CI 0.42-0.87). The misclassification rate varied with cutoff, and ranged from 0-83% and 0-58% using upper limit of normal (ULN) and twice ULN, respectively. The within patient variability was 0.37 for the entire cohort, and 0.33 when three patients with intercurrent acute inflammation were excluded. CRP exhibits short term variability in HD patients, resulting in a risk of misclassification depending on sampling time and chosen cutoff point. Single CRP values must be interpreted with caution, and multiple measurements, or use of other biomarkers, should be considered.

  19. Methadone maintenance.

    PubMed

    Russell-Taylor, W J

    1975-07-01

    Management of heroin addiction with oral methadone continues to create controversy. Since the first Dole-Nyswander experiments were reported almost a decade ago, methadone maintenance programs have been started in every major city in North America at an estimated public cost in the hundreds of millions of tax dollars. By the end of 1972, the United States alone spending at an annual rate in excess of $250 million, and an expenditure rate in excess of one billion dollars was occurring by the end of 1973. Despite the FDA investigational New Drug status of methadone when used for this purpose, objective data have still not been generated to substantiate the worth of Narcotic Substitution Therapy. Iatrogenic consequences are on the increase, and scepticism is growing among most pharmacologists. A critical evaluation of the entire methadone maintenance approach is being advocated by many and being attempted by some. Data currently being generated at the Philadelphia General Hospital suggest that many patients may have been better managed without methadone.

  20. Zebrafish pronephros tubulogenesis and epithelial identity maintenance are reliant on the polarity proteins Prkc iota and zeta

    PubMed Central

    Gerlach, Gary F.; Wingert, Rebecca A.

    2014-01-01

    The zebrafish pronephros provides an excellent in vivo system to study the mechanisms of vertebrate nephron development. When and how renal progenitors in the zebrafish embryo undergo tubulogenesis to form nephrons is poorly understood, but is known to involve a mesenchymal to epithelial transition (MET) and the acquisition of polarity. Here, we determined the precise timing of these events in pronephros tubulogenesis. As the ternary polarity complex is an essential regulator of epithelial cell polarity across tissues, we performed gene knockdown studies to assess the roles of the related factors atypical protein kinase C iota and zeta (prkcι, prkcζ). We found that prkcι and prkcζ serve partially redundant functions to establish pronephros tubule epithelium polarity. Further, the loss of prkcι or the combined knockdown of prkcι/ζ disrupted proximal tubule morphogenesis and podocyte migration due to cardiac defects that prevented normal fluid flow to the kidney. Surprisingly, tubule cells in prkcι/ζ morphants displayed ectopic expression of the transcription factor pax2a and the podocyte-associated genes wt1a, wt1b, and podxl, suggesting that prkcι/ζ are needed to maintain renal epithelial identity. Knockdown of genes essential for cardiac contractility and vascular flow to the kidney, such as tnnt2a, or elimination of pronephros fluid output through knockdown of the intraflagellar transport gene ift88, was not associated with ectopic pronephros gene expression, thus suggesting a unique role for prkcι/ζ in maintaining tubule epithelial identity separate from the consequence of disruptions to renal fluid flow. Interestingly, knockdown of pax2a, but not wt1a, was sufficient to rescue ectopic tubule gene expression in prkcι/ζ morphants. These data suggest a model in which the redundant activities of prkcι and prkcζ are essential to establish tubule epithelial polarity and also serve to maintain proper epithelial cell type identity in the tubule by

  1. Zebrafish pronephros tubulogenesis and epithelial identity maintenance are reliant on the polarity proteins Prkc iota and zeta.

    PubMed

    Gerlach, Gary F; Wingert, Rebecca A

    2014-12-15

    The zebrafish pronephros provides an excellent in vivo system to study the mechanisms of vertebrate nephron development. When and how renal progenitors in the zebrafish embryo undergo tubulogenesis to form nephrons is poorly understood, but is known to involve a mesenchymal to epithelial transition (MET) and the acquisition of polarity. Here, we determined the precise timing of these events in pronephros tubulogenesis. As the ternary polarity complex is an essential regulator of epithelial cell polarity across tissues, we performed gene knockdown studies to assess the roles of the related factors atypical protein kinase C iota and zeta (prkcι, prkcζ). We found that prkcι and prkcζ serve partially redundant functions to establish pronephros tubule epithelium polarity. Further, the loss of prkcι or the combined knockdown of prkcι/ζ disrupted proximal tubule morphogenesis and podocyte migration due to cardiac defects that prevented normal fluid flow to the kidney. Surprisingly, tubule cells in prkcι/ζ morphants displayed ectopic expression of the transcription factor pax2a and the podocyte-associated genes wt1a, wt1b, and podxl, suggesting that prkcι/ζ are needed to maintain renal epithelial identity. Knockdown of genes essential for cardiac contractility and vascular flow to the kidney, such as tnnt2a, or elimination of pronephros fluid output through knockdown of the intraflagellar transport gene ift88, was not associated with ectopic pronephros gene expression, thus suggesting a unique role for prkcι/ζ in maintaining tubule epithelial identity separate from the consequence of disruptions to renal fluid flow. Interestingly, knockdown of pax2a, but not wt1a, was sufficient to rescue ectopic tubule gene expression in prkcι/ζ morphants. These data suggest a model in which the redundant activities of prkcι and prkcζ are essential to establish tubule epithelial polarity and also serve to maintain proper epithelial cell type identity in the tubule by

  2. A critical role for the protein kinase PKK in the maintenance of recirculating mature B cells and the development of B1 cells.

    PubMed

    Chen, Luojing; Oleksyn, David; Pulvino, Mary; Sanz, Ignacio; Ryan, Daniel; Ryan, Charlotte; Lin, Chyuan-Sheng; Poligone, Brian; Pentland, Alice P; Ritchlin, Christopher; Zhao, Jiyong

    2016-04-01

    Protein kinase C associated kinase (PKK) regulates NF-κB activation and is required for the survival of certain lymphoma cells. Mice lacking PKK die soon after birth, and previous studies suggest that the role of PKK in B cell development might be context dependent. We have generated a mouse strain harboring conditional null alleles for PKK and a Cre-recombinase transgene under the control of the endogenous CD19 promoter. In the present study, we show that knockout of PKK in B cells results in the reduction of long-lived recirculating mature B cell population in lymph nodes and bone marrow as well as a decrease in peritoneal B1 cells, while PKK deficiency has no apparent effect on early B cell development in bone marrow or the development of follicular and marginal zone B cells in the spleen. In addition, we demonstrate that PKK-deficient B cells display defective proliferation and survival responses to stimulation of B cell receptor (BCR), which may underlie the reduction of recirculating mature B cells in PKK mutant mice. Consistently, BCR-mediated NF-κB activation, known to be required for the survival of activated but not resting B cells, is attenuated in PKK-deficient B cells. Thus, our results reveal a critical role of PKK in the maintenance of recirculating mature B cells as well as the development of B1 cells in mice.

  3. Structural Basis for Dimer Formation of Human Condensin Structural Maintenance of Chromosome Proteins and Its Implications for Single-stranded DNA Recognition*

    PubMed Central

    Uchiyama, Susumu; Kawahara, Kazuki; Hosokawa, Yuki; Fukakusa, Shunsuke; Oki, Hiroya; Nakamura, Shota; Kojima, Yukiko; Noda, Masanori; Takino, Rie; Miyahara, Yuya; Maruno, Takahiro; Kobayashi, Yuji; Ohkubo, Tadayasu; Fukui, Kiichi

    2015-01-01

    Eukaryotic structural maintenance of chromosome proteins (SMC) are major components of cohesin and condensins that regulate chromosome structure and dynamics during cell cycle. We here determine the crystal structure of human condensin SMC hinge heterodimer with ∼30 residues of coiled coils. The structure, in conjunction with the hydrogen exchange mass spectrometry analyses, revealed the structural basis for the specific heterodimer formation of eukaryotic SMC and that the coiled coils from two different hinges protrude in the same direction, providing a unique binding surface conducive for binding to single-stranded DNA. The characteristic hydrogen exchange profiles of peptides constituted regions especially across the hinge-hinge dimerization interface, further suggesting the structural alterations upon single-stranded DNA binding and the presence of a half-opened state of hinge heterodimer. This structural change potentially relates to the DNA loading mechanism of SMC, in which the hinge domain functions as an entrance gate as previously proposed for cohesin. Our results, however, indicated that this is not the case for condensins based on the fact that the coiled coils are still interacting with each other, even when DNA binding induces structural changes in the hinge region, suggesting the functional differences of SMC hinge domain between condensins and cohesin in DNA recognition. PMID:26491021

  4. Alignment of Homologous Chromosomes and Effective Repair of Programmed DNA Double-Strand Breaks during Mouse Meiosis Require the Minichromosome Maintenance Domain Containing 2 (MCMDC2) Protein.

    PubMed

    Finsterbusch, Friederike; Ravindranathan, Ramya; Dereli, Ihsan; Stanzione, Marcello; Tränkner, Daniel; Tóth, Attila

    2016-10-01

    Orderly chromosome segregation during the first meiotic division requires meiotic recombination to form crossovers between homologous chromosomes (homologues). Members of the minichromosome maintenance (MCM) helicase family have been implicated in meiotic recombination. In addition, they have roles in initiation of DNA replication, DNA mismatch repair and mitotic DNA double-strand break repair. Here, we addressed the function of MCMDC2, an atypical yet conserved MCM protein, whose function in vertebrates has not been reported. While we did not find an important role for MCMDC2 in mitotically dividing cells, our work revealed that MCMDC2 is essential for fertility in both sexes due to a crucial function in meiotic recombination. Meiotic recombination begins with the introduction of DNA double-strand breaks into the genome. DNA ends at break sites are resected. The resultant 3-prime single-stranded DNA overhangs recruit RAD51 and DMC1 recombinases that promote the invasion of homologous duplex DNAs by the resected DNA ends. Multiple strand invasions on each chromosome promote the alignment of homologous chromosomes, which is a prerequisite for inter-homologue crossover formation during meiosis. We found that although DNA ends at break sites were evidently resected, and they recruited RAD51 and DMC1 recombinases, these recombinases were ineffective in promoting alignment of homologous chromosomes in the absence of MCMDC2. Consequently, RAD51 and DMC1 foci, which are thought to mark early recombination intermediates, were abnormally persistent in Mcmdc2-/- meiocytes. Importantly, the strand invasion stabilizing MSH4 protein, which marks more advanced recombination intermediates, did not efficiently form foci in Mcmdc2-/- meiocytes. Thus, our work suggests that MCMDC2 plays an important role in either the formation, or the stabilization, of DNA strand invasion events that promote homologue alignment and provide the basis for inter-homologue crossover formation during

  5. Biphasic response of checkpoint control proteins in hyperoxia: exposure to lower levels of oxygen induces genome maintenance genes in experimental baboon BPD.

    PubMed

    Das, Kumuda C; Wasnick, John D

    2014-10-01

    Breathing high concentrations of oxygen (hyperoxia) causes lung injury and is associated with lung diseases such as bronchopulmonary dysplasia (BPD), respiratory distress syndrome and persistent pulmonary hypertension of the newborns. Hyperoxia (95-100 %O2) causes DNA damage and growth arrest of lung cells and consequently cells die by apoptosis or necrosis. Although supplemental oxygen therapy is clinically important, the level and duration of hyperoxic exposure that would allow lung cells to reenter the cell cycle remains unclear. We hypothesized that cells exposed to lower concentrations of hyperoxia will retain the capacity to enter cell cycle when recovered in room air. We employed varying concentrations of oxygen (21-95 %) to determine the response of lung cells to hyperoxia. Our results indicate that cells were growth arrested and failed to reenter the cell cycle when exposed to greater than 60 % oxygen. Cell cycle checkpoint proteins were increased in a biphasic manner, increasing until 70 % oxygen, but declined in greater than 90 % oxygen. Microarray analysis shows that there is significant decrease in the abundance of Cdks 6-8 and retinoblastoma protein (Rb), p107 and p130 in exposure to 90 % oxygen for 48 h. We further tested the effect of clinically relevant as needed oxygen [(pro-re-nata (prn)] in premature infant (125-days and 140-days) baboon model of BPD. The microarray results show that 6 or 14d PRN oxygen-exposed animals had induced expression of chromosomal maintenance genes (MCMs), genes related to anti-inflammation, proliferation, and differentiation.

  6. Methyl-CpG-binding protein MBD2 plays a key role in maintenance and spread of DNA methylation at CpG islands and shores in cancer.

    PubMed

    Stirzaker, C; Song, J Z; Ng, W; Du, Q; Armstrong, N J; Locke, W J; Statham, A L; French, H; Pidsley, R; Valdes-Mora, F; Zotenko, E; Clark, S J

    2017-03-01

    Cancer is characterised by DNA hypermethylation and gene silencing of CpG island-associated promoters, including tumour-suppressor genes. The methyl-CpG-binding domain (MBD) family of proteins bind to methylated DNA and can aid in the mediation of gene silencing through interaction with histone deacetylases and histone methyltransferases. However, the mechanisms responsible for eliciting CpG island hypermethylation in cancer, and the potential role that MBD proteins play in modulation of the methylome remain unclear. Our previous work demonstrated that MBD2 preferentially binds to the hypermethylated GSTP1 promoter CpG island in prostate cancer cells. Here, we use functional genetic approaches to investigate if MBD2 plays an active role in reshaping the DNA methylation landscape at this locus and genome-wide. First, we show that loss of MBD2 results in inhibition of both maintenance and spread of de novo methylation of a transfected construct containing the GSTP1 promoter CpG island in prostate cancer cells and Mbd2-/- mouse fibroblasts. De novo methylation was rescued by transient expression of Mbd2 in Mbd2-/- cells. Second, we show that MBD2 depletion triggers significant hypomethylation genome-wide in prostate cancer cells with concomitant loss of MBD2 binding at promoter and enhancer regulatory regions. Finally, CpG islands and shores that become hypomethylated after MBD2 depletion in LNCaP cancer cells show significant hypermethylation in clinical prostate cancer samples, highlighting a potential active role of MBD2 in promoting cancer-specific hypermethylation. Importantly, co-immunoprecipiation of MBD2 shows that MBD2 associates with DNA methyltransferase enzymes 1 and 3A. Together our results demonstrate that MBD2 has a critical role in 'rewriting' the cancer methylome at specific regulatory regions.

  7. The role of Schizosaccharomyces pombe Rad32, the Mre11 homologue, and other DNA damage response proteins in non-homologous end joining and telomere length maintenance.

    PubMed Central

    Wilson, S; Warr, N; Taylor, D L; Watts, F Z

    1999-01-01

    The Schizosaccharomyces pombe homologue of Mre11, Rad32, is required for repair of UV- and ionising radiation-induced DNA damage and meiotic recombination. In this study we have investigated the role of Rad32 and other DNA damage response proteins in non-homologous end joining (NHEJ) and telomere length maintenance in S.pombe. We show that NHEJ in S.pombe occurs by an error-prone mechanism, in contrast to the accurate repair observed in Saccharomyces cerevisiae. Deletion of the rad32 gene results in a modest reduction in NHEJ activity and the remaining repair events that occur are accurate. Mutations in two of the phosphoesterase motifs in Rad32 have no effect on the efficiency or accuracy of end joining, suggesting that the role of Rad32 protein may be to recruit another nuclease(s) for processing during the end joining reaction. We also analysed NHEJ in other DNA damage response mutants and showed that the checkpoint mutant rad3-d and two recombination mutants defective in rhp51 and rhp54 (homologues of S.cerevisiae RAD51 and RAD54, respectively) are not affected. However disruption of rad22, rqh1 and rhp9 / crb2 (homologues of the S.cerevisiae RAD52, SGS1 and RAD9 genes) resulted in increased NHEJ activity. Telomere lengths in the rad32, rhp9 and rqh1 null alleles were reduced to varying extents intermediate between the lengths observed in wild-type and rad3 null cells. PMID:10373582

  8. Alignment of Homologous Chromosomes and Effective Repair of Programmed DNA Double-Strand Breaks during Mouse Meiosis Require the Minichromosome Maintenance Domain Containing 2 (MCMDC2) Protein

    PubMed Central

    Ravindranathan, Ramya; Dereli, Ihsan; Stanzione, Marcello; Tóth, Attila

    2016-01-01

    Orderly chromosome segregation during the first meiotic division requires meiotic recombination to form crossovers between homologous chromosomes (homologues). Members of the minichromosome maintenance (MCM) helicase family have been implicated in meiotic recombination. In addition, they have roles in initiation of DNA replication, DNA mismatch repair and mitotic DNA double-strand break repair. Here, we addressed the function of MCMDC2, an atypical yet conserved MCM protein, whose function in vertebrates has not been reported. While we did not find an important role for MCMDC2 in mitotically dividing cells, our work revealed that MCMDC2 is essential for fertility in both sexes due to a crucial function in meiotic recombination. Meiotic recombination begins with the introduction of DNA double-strand breaks into the genome. DNA ends at break sites are resected. The resultant 3-prime single-stranded DNA overhangs recruit RAD51 and DMC1 recombinases that promote the invasion of homologous duplex DNAs by the resected DNA ends. Multiple strand invasions on each chromosome promote the alignment of homologous chromosomes, which is a prerequisite for inter-homologue crossover formation during meiosis. We found that although DNA ends at break sites were evidently resected, and they recruited RAD51 and DMC1 recombinases, these recombinases were ineffective in promoting alignment of homologous chromosomes in the absence of MCMDC2. Consequently, RAD51 and DMC1 foci, which are thought to mark early recombination intermediates, were abnormally persistent in Mcmdc2-/- meiocytes. Importantly, the strand invasion stabilizing MSH4 protein, which marks more advanced recombination intermediates, did not efficiently form foci in Mcmdc2-/- meiocytes. Thus, our work suggests that MCMDC2 plays an important role in either the formation, or the stabilization, of DNA strand invasion events that promote homologue alignment and provide the basis for inter-homologue crossover formation during

  9. WD40-repeat protein 62 is a JNK-phosphorylated spindle pole protein required for spindle maintenance and timely mitotic progression

    PubMed Central

    Bogoyevitch, Marie A.; Yeap, Yvonne Y. C.; Qu, Zhengdong; Ngoei, Kevin R.; Yip, Yan Y.; Zhao, Teresa T.; Heng, Julian I.; Ng, Dominic C. H.

    2012-01-01

    Summary The impact of aberrant centrosomes and/or spindles on asymmetric cell division in embryonic development indicates the tight regulation of bipolar spindle formation and positioning that is required for mitotic progression and cell fate determination. WD40-repeat protein 62 (WDR62) was recently identified as a spindle pole protein linked to the neurodevelopmental defect of microcephaly but its roles in mitosis have not been defined. We report here that the in utero electroporation of neuroprogenitor cells with WDR62 siRNAs induced their cell cycle exit and reduced their proliferative capacity. In cultured cells, we demonstrated cell-cycle-dependent accumulation of WDR62 at the spindle pole during mitotic entry that persisted until metaphase–anaphase transition. Utilizing siRNA depletion, we revealed WDR62 function in stabilizing the mitotic spindle specifically during metaphase. WDR62 loss resulted in spindle orientation defects, decreased the integrity of centrosomes displaced from the spindle pole and delayed mitotic progression. Additionally, we revealed JNK phosphorylation of WDR62 is required for maintaining metaphase spindle organization during mitosis. Our study provides the first functional characterization of WDR62 and has revealed requirements for JNK/WDR62 signaling in mitotic spindle regulation that may be involved in coordinating neurogenesis. PMID:22899712

  10. WD40-repeat protein 62 is a JNK-phosphorylated spindle pole protein required for spindle maintenance and timely mitotic progression.

    PubMed

    Bogoyevitch, Marie A; Yeap, Yvonne Y C; Qu, Zhengdong; Ngoei, Kevin R; Yip, Yan Y; Zhao, Teresa T; Heng, Julian I; Ng, Dominic C H

    2012-11-01

    The impact of aberrant centrosomes and/or spindles on asymmetric cell division in embryonic development indicates the tight regulation of bipolar spindle formation and positioning that is required for mitotic progression and cell fate determination. WD40-repeat protein 62 (WDR62) was recently identified as a spindle pole protein linked to the neurodevelopmental defect of microcephaly but its roles in mitosis have not been defined. We report here that the in utero electroporation of neuroprogenitor cells with WDR62 siRNAs induced their cell cycle exit and reduced their proliferative capacity. In cultured cells, we demonstrated cell-cycle-dependent accumulation of WDR62 at the spindle pole during mitotic entry that persisted until metaphase-anaphase transition. Utilizing siRNA depletion, we revealed WDR62 function in stabilizing the mitotic spindle specifically during metaphase. WDR62 loss resulted in spindle orientation defects, decreased the integrity of centrosomes displaced from the spindle pole and delayed mitotic progression. Additionally, we revealed JNK phosphorylation of WDR62 is required for maintaining metaphase spindle organization during mitosis. Our study provides the first functional characterization of WDR62 and has revealed requirements for JNK/WDR62 signaling in mitotic spindle regulation that may be involved in coordinating neurogenesis.

  11. High-sensitivity C-reactive protein is predictive of successful cardioversion for atrial fibrillation and maintenance of sinus rhythm after conversion.

    PubMed

    Watanabe, Eiichi; Arakawa, Tomoharu; Uchiyama, Tatsushi; Kodama, Itsuo; Hishida, Hitoshi

    2006-04-14

    Cardioversion for atrial fibrillation (AF) is the most effective treatment for the restoration of sinus rhythm (SR). Recently, an elevated level of hs-CRP has been shown to be associated with AF burden, suggesting that inflammation increases the propensity for persistence of AF. We examined whether the level of high-sensitivity C-reactive protein (hs-CRP) was predictive of the outcome of cardioversion for AF. One hundred and six patients with a history of symptomatic AF lasting > or =1 day (age 63+/-14 years, mean+/-S.D.) underwent cardioversion. Echocardiography and hs-CRP assay were performed immediately prior to cardioversion. SR was restored in 84 patients (79%). By using selected cutoff values, multiple discriminant analysis revealed significant associations between successful cardioversion and a shorter duration of AF (AF duration< or =36 days, odds ratio (OR), 0.98; 95% confidence interval (CI), 0.97-0.99), smaller left atrial diameter (left atrial diameter< or =40 mm, OR 0.82, 95% CI 0.71-0.94), better-preserved left ventricular ejection fraction (left ventricular ejection fraction> or =60%, OR 0.92, 95% CI 0.86-0.99), and lower hs-CRP level (hs-CRP< or =0.12 mg/dL, OR 0.33, 95% CI 0.21-0.51). During a follow-up period of 140+/-144 days, AF recurred in 64 patients (76%). By using a cutoff value of hs-CRP> or =0.06 mg/dL, Cox proportional-hazards regression model found that only hs-CRP level was an independent predictor of AF recurrence (OR 5.30, 95% CI 2.46-11.5) after adjustment for coexisting cardiovascular risks. When patients were divided by the hs-CRP level of 0.06 mg/dL, percentage of maintenance of SR below and above the cutoff was 53% and 4%, respectively (log-rank test, p<0.0001). hs-CRP level determined prior to cardioversion represents an independent predictor of both successful cardioversion for AF and the maintenance of SR after conversion.

  12. Protein supplements after weight loss do not improve weight maintenance compared with recommended dietary protein intake despite beneficial effects on appetite sensation and energy expenditure: a randomized, controlled, double-blinded trial.

    PubMed

    Kjølbæk, Louise; Sørensen, Lone Brinkmann; Søndertoft, Nadja Buus; Rasmussen, Carrie Klestrup; Lorenzen, Janne Kunchel; Serena, Anja; Astrup, Arne; Larsen, Lesli Hingstrup

    2017-08-01

    Background: High-protein diets increase weight loss (WL) during energy restriction; therefore, it has been suggested that additional protein intake may improve weight maintenance (WM) after WL.Objective: We investigated the effect of protein supplements from either whey with or without calcium or soy on WM success after WL compared with that of a control.Design: In a randomized, controlled, double-blinded trial, 220 participants aged 18-60 y with body mass index (in kg/m(2)) from 27.6 to 40.4 were included. The study was initiated with an 8-wk WL period followed by a 24-wk WM period. During WM, participants consumed the following isocaloric supplements (45-48 g/d): whey and calcium (whey+), whey, soy, or maltodextrin (control). Data were collected at baseline, before WM, and after WM (weeks 0, 8, and 32, respectively) and included body composition, blood biochemistry, and blood pressure. Meal tests were performed to investigate diet-induced-thermogenesis (DIT) and appetite sensation. Compliance was tested by 24-h urinary nitrogen excretion.Results: A total of 151 participants completed the WM period. The control and 3 protein supplements did not result in different mean ± SD weight regains (whey+: 2.19 ± 4.6 kg; whey: 2.01 ± 4.6 kg; soy: 1.76 ± 4.7 kg; and control: 2.23 ± 3.8 kg; P = 0.96), fat mass regains (whey+: 0.46 ± 4.5 kg; whey: 0.11 ± 4.1 kg; soy: 0.15 ± 4.1 kg; and control: 0.54 ± 3.3 kg; P = 0.96), or improvements in lean body mass (whey+: 1.87 ± 1.7 kg; whey: 1.94 ± 1.3 kg; soy: 1.58 ± 1.4 kg; and control: 1.74 ± 1.4 kg; P = 0.50) during WM. Changes in blood pressure and blood biochemistry were not different between groups. Compared with the control, protein supplementation resulted in higher DIT (∼30 kJ/2.5 h) and resting energy expenditure (243 kJ/d) and an anorexigenic appetite-sensation profile.Conclusion: Protein supplementation does not result in improved WM success, or blood biochemistry after WL compared with the effects of normal

  13. Effects of Recombinant Human Bone Morphogenetic Protein-2 Dose and Ceramic Composition on New Bone Formation and Space Maintenance in a Canine Mandibular Ridge Saddle Defect Model

    PubMed Central

    Talley, Anne D.; Kalpakci, Kerem N.; Shimko, Daniel A.; Zienkiewicz, Katarzyna J.; Cochran, David L.

    2016-01-01

    Treatment of mandibular osseous defects is a significant clinical challenge. Maintenance of the height and width of the mandibular ridge is essential for placement of dental implants and restoration of normal dentition. While guided bone regeneration using protective membranes is an effective strategy for maintaining the anatomic contour of the ridge and promoting new bone formation, complications have been reported, including wound failure, seroma, and graft exposure leading to infection. In this study, we investigated injectable low-viscosity (LV) polyurethane/ceramic composites augmented with 100 μg/mL (low) or 400 μg/mL (high) recombinant human bone morphogenetic protein-2 (rhBMP-2) as space-maintaining bone grafts in a canine mandibular ridge saddle defect model. LV grafts were injected as a reactive paste that set in 5–10 min to form a solid porous composite with bulk modulus exceeding 1 MPa. We hypothesized that compression-resistant LV grafts would enhance new bone formation and maintain the anatomic contour of the mandibular ridge without the use of protective membranes. At the rhBMP-2 dose recommended for the absorbable collagen sponge carrier in dogs (400 μg/mL), LV grafts maintained the width and height of the host mandibular ridge and supported new bone formation, while at suboptimal (100 μg/mL) doses, the anatomic contour of the ridge was not maintained. These findings indicate that compression-resistant bone grafts with bulk moduli exceeding 1 MPa and rhBMP-2 doses comparable to that recommended for the collagen sponge carrier support new bone formation and maintain ridge height and width in mandibular ridge defects without protective membranes. PMID:26800574

  14. Minichromosome Maintenance Protein 7 is a potential therapeutic target in human cancer and a novel prognostic marker of non-small cell lung cancer

    PubMed Central

    2011-01-01

    Background The research emphasis in anti-cancer drug discovery has always been to search for a drug with the greatest antitumor potential but fewest side effects. This can only be achieved if the drug used is against a specific target located in the tumor cells. In this study, we evaluated Minichromosome Maintenance Protein 7 (MCM7) as a novel therapeutic target in cancer. Results Immunohistochemical analysis showed that MCM7 was positively stained in 196 of 331 non-small cell lung cancer (NSCLC), 21 of 29 bladder tumor and 25 of 70 liver tumor cases whereas no significant staining was observed in various normal tissues. We also found an elevated expression of MCM7 to be associated with poor prognosis for patients with NSCLC (P = 0.0055). qRT-PCR revealed a higher expression of MCM7 in clinical bladder cancer tissues than in corresponding non-neoplastic tissues (P < 0.0001), and we confirmed that a wide range of cancers also overexpressed MCM7 by cDNA microarray analysis. Suppression of MCM7 using specific siRNAs inhibited incorporation of BrdU in lung and bladder cancer cells overexpressing MCM7, and suppressed the growth of those cells more efficiently than that of normal cell strains expressing lower levels of MCM7. Conclusions Since MCM7 expression was generally low in a number of normal tissues we examined, MCM7 has the characteristics of an ideal candidate for molecular targeted cancer therapy in various tumors and also as a good prognostic biomarker for NSCLC patients. PMID:21619671

  15. Maintenance of coat protein N-terminal net charge and not primary sequence is essential for zucchini yellow mosaic virus systemic infectivity.

    PubMed

    Kimalov, Boaz; Gal-On, Amit; Stav, Ran; Belausov, Eduard; Arazi, Tzahi

    2004-11-01

    Zucchini yellow mosaic virus (ZYMV) surface exposed coat protein (CP) N-terminal domain (Nt) is 43 aa long and contains an equal number of positively and negatively charged amino acid residues (CP-Nt net charge = 0). A ZYMV-AGII truncation mutant lacking the first 20 aa of its CP-Nt (AGII-CP Delta 20; CP-Nt net charge = +2) was found to be systemically non-infectious even though AGII mutants harbouring larger CP-Nt deletions were previously demonstrated to be fully infectious. Nevertheless, AGII-CP Delta 20 infectivity was restored by fusion to its CP-Nt two Asp residues or a negatively charged Myc peptide, both predicted to neutralize CP-Nt net positive charge. To evaluate further the significance of CP-Nt net charge for AGII infectivity, a series of CP-Nt net charge mutants was generated and analysed for systemic infectivity of squash plants. AGII-CP(KKK) harbouring a CP-Nt amino fusion of three Lys residues (CP-Nt net charge = +3) was not systemically infectious. Addition of up to four Asp residues to CP-Nt did not abolish virus infectivity, although certain mutants were genetically unstable and had delayed infectivity. Addition of five negatively charged residues abolished infectivity (AGII-CP(DDDDD); CP-Nt net charge = -5) even though a recombinant CP(DDDDD) could assemble into potyviral-like particle in bacteria. Neutralization of CP-Nt net charge by fusing Asp or Lys residues recovered infectivity of AGII-CP(KKK) and AGII-CP(DDDDD). GFP-tagging of these mutants has demonstrated that both viruses have defective cell-to-cell movement. Together, these findings suggest that maintenance of CP-Nt net charge and not primary sequence is essential for ZYMV infectivity.

  16. Effect of different feeding regimens on energy and protein utilization and partitioning for maintenance and growth in pre-weaned lambs reared artificially.

    PubMed

    Danso, A S; Morel, P C H; Kenyon, P R; Blair, H T

    2016-12-01

    Estimation of metabolizable energy (ME) requirement for maintenance (ME) and growth (ME) in pre-weaned lambs have been limited to milk-only fed lambs. This study aimed to determine energy and nitrogen metabolisability of milk and pellets when fed together, compare the growth and chemical body composition of lambs fed varying levels of pellets in addition to milk, and to estimate ME, ME, and the CP:ME ratio requirements for growth. The study included 32 twin-born Romney-cross ram lambs. Four lambs were slaughtered at 24 h post-partum to estimate initial body composition and the remaining 28 were assigned to 1 of 4 treatment groups of 7. Group 1 was fed milk replacer (MR) only; group 2 was fed MR and allowed ad libitum access to pellets; groups 3 and 4 were offered 30% and 60%, respectively of the average pellet intake of the ad libitum group the previous day while being fed MR. Milk replacer was fed as a proportion of the lamb's live weight (LW). Lambs from each treatment were placed in metabolic cages at 17 kg LW for 4 d to allow for total fecal and urine collection. All lambs were slaughtered at 18 kg LW. The ADG, ADG:ME ratio, stomach and liver weight, and rumen papillae lengths increased ( < 0.05) with increasing pellet intake. Increasing daily ME intake increased ( < 0.05) both daily energy and protein deposition but had no effect ( > 0.05) on fat deposition. However, the total chemical body composition was unaffected ( > 0.05) by dietary treatment. Digestibility of energy and N decreased ( < 0.05) with increasing ME intake. Percent energy and N retained for growth were 96% vs. 71% and 72% vs. 30% for milk and pellets, respectively. The ME and ME values obtained were 0.40 MJ ME/kg LW·d and 13.8 MJ ME/kg ADG, respectively. The CP:ME ratio of MR and pellet was 11.1 and 15.7, respectively. However, a simulation model suggested that lambs require a CP:ME ratio of 13.1 at 5 kg and 10.9 at 18 kg LW, indicating that protein intake may be limiting to lamb growth in

  17. Maintenance Business Plans.

    ERIC Educational Resources Information Center

    Adams, Matt

    2002-01-01

    Discusses maintenance business plans, statements which provide accountability for facilities maintenance organizations' considerable budgets. Discusses the plan's components: statement of plan objectives, macro and detailed description of the facility assets, maintenance function descriptions, description of key performance indicators, milestone…

  18. Maintenance Management Update.

    ERIC Educational Resources Information Center

    Sternloff, Robert E.

    1987-01-01

    Current trends in park maintenance are overviewed, including maintenance impact statements, avoidance of cost through efficient use and national resource conservation, horticultural accomplishments that influence maintenance management, and vandalism prevention. (CB)

  19. Maintenance Management Update.

    ERIC Educational Resources Information Center

    Sternloff, Robert E.

    1987-01-01

    Current trends in park maintenance are overviewed, including maintenance impact statements, avoidance of cost through efficient use and national resource conservation, horticultural accomplishments that influence maintenance management, and vandalism prevention. (CB)

  20. Human Cytomegalovirus IE2 Protein Disturbs Brain Development by the Dysregulation of Neural Stem Cell Maintenance and the Polarization of Migrating Neurons.

    PubMed

    Han, Dasol; Byun, Sung-Hyun; Kim, Juwan; Kwon, Mookwang; Pleasure, Samuel J; Ahn, Jin-Hyun; Yoon, Keejung

    2017-09-01

    Despite the high incidence of severe defects in the central nervous system caused by human cytomegalovirus (HCMV) congenital infection, the mechanism of HCMV neuropathogenesis and the roles of individual viral genes have not yet been fully determined. In this study, we show that the immediate-early 2 (IE2) protein may play a key role in HCMV-caused neurodevelopmental disorders. IE2-transduced neural progenitor cells gave rise to neurospheres with a lower frequency and produced smaller neurospheres than control cells in vitro, indicating reduction of self-renewal and expansion of neural progenitors by IE2. At 2 days after in utero electroporation into the ventricle of the developing brain, a dramatically lower percentage of IE2-expressing cells was detected in the ventricular zone (VZ) and cortical plate (CP) compared to control cells, suggesting that IE2 concurrently dysregulates neural stem cell maintenance in the VZ and neuronal migration to the CP. In addition, most IE2(+) cells in the lower intermediate zone either showed multipolar morphology with short neurites or possessed nonradially oriented processes, whereas control cells had long, radially oriented monopolar or bipolar neurites. IE2(+) callosal axons also failed to cross the midline to form the corpus callosum. Furthermore, we provide molecular evidence that the cell cycle arrest and DNA binding activities of IE2 appear to be responsible for the increased neural stem cell exit from the VZ and cortical migrational defects, respectively. Collectively, our results demonstrate that IE2 disrupts the orderly process of brain development in a stepwise manner to further our understanding of neurodevelopmental HCMV pathogenesis.IMPORTANCE HCMV brain pathogenesis has been studied in limited experimental settings, such as in vitro HCMV infection of neural progenitor cells or in vivo murine CMV infection of the mouse brain. Here, we show that IE2 is a pivotal factor that contributes to HCMV-induced abnormalities in

  1. A Mitogen-Activated Protein Kinase Tmk3 Participates in High Osmolarity Resistance, Cell Wall Integrity Maintenance and Cellulase Production Regulation in Trichoderma reesei

    PubMed Central

    Wang, Mingyu; Zhao, Qiushuang; Yang, Jinghua; Jiang, Baojie; Wang, Fangzhong; Liu, Kuimei; Fang, Xu

    2013-01-01

    The mitogen-activated protein kinase (MAPK) pathways are important signal transduction pathways conserved in essentially all eukaryotes, but haven't been subjected to functional studies in the most important cellulase-producing filamentous fungus Trichoderma reesei. Previous reports suggested the presence of three MAPKs in T. reesei: Tmk1, Tmk2, and Tmk3. By exploring the phenotypic features of T. reesei Δtmk3, we first showed elevated NaCl sensitivity and repressed transcription of genes involved in glycerol/trehalose biosynthesis under higher osmolarity, suggesting Tmk3 participates in high osmolarity resistance via derepression of genes involved in osmotic stabilizer biosynthesis. We also showed significant downregulation of genes encoding chitin synthases and a β-1,3-glucan synthase, decreased chitin content, ‘budded’ hyphal appearance typical to cell wall defective strains, and increased sensitivity to calcofluor white/Congo red in the tmk3 deficient strain, suggesting Tmk3 is involved in cell wall integrity maintenance in T. reesei. We further observed the decrease of cellulase transcription and production in T. reesei Δtmk3 during submerged cultivation, as well as the presence of MAPK phosphorylation sites on known transcription factors involved in cellulase regulation, suggesting Tmk3 is also involved in the regulation of cellulase production. Finally, the expression of cell wall integrity related genes, the expression of cellulase coding genes, cellulase production and biomass accumulation were compared between T. reesei Δtmk3 grown in solid state media and submerged media, showing a strong restoration effect in solid state media from defects resulted from tmk3 deletion. These results showed novel physiological processes that fungal Hog1-type MAPKs are involved in, and present the first experimental investigation of MAPK signaling pathways in T. reesei. Our observations on the restoration effect during solid state cultivation suggest that T. reesei

  2. Histopathological variables and biomarkers enhancer of zeste homologue 2, Ki-67 and minichromosome maintenance protein 7 as prognosticators in primarily endocrine-treated prostate cancer.

    PubMed

    Tolonen, Teemu T; Tammela, Teuvo L J; Kujala, Paula M; Tuominen, Vilppu J; Isola, Jorma J; Visakorpi, Tapio

    2011-11-01

    • To evaluate the prognostic value of histopathological variables and immunostainings of biomarkers enhancer of zeste homologue 2 (EZH2), Ki-67 and minichromosome maintenance protein 7 (MCM7) from core biopsies of hormonally treated patients with prostate cancer. • Biopsies of 247 primarily endocrine-treated patients were analysed for histopathological characteristics and Gleason scores (GS) according to the revised guidelines of International Society of Urologic Pathology (ISUP) consensus conference 2005. • Immunohistochemical stainings were analysed with the aid of digital image analysis. • The prognostic value of the histopathological variables and the biomarkers was analysed with univariate and multivariate Cox regression analysis, with biochemical recurrence as an endpoint. • Biomarkers EZH2 (relative risk [RR] 2.0, 95% confidence interval 1.2-3.3), Ki-67 (3.4, 2.1-5.5) and MCM7 (2.4, 1.5-3.9) were significantly associated with progression-free survival in a univariate analysis. • Ki-67 immunostaining index detected high-risk patients with GS of 7 (9.1, 8.0-10.3). • In a multivariate analysis with non-conventional GS groups 5-7 (3 + 4), 7(4 + 3)-8, and 9-10, the independent prognostic markers were pretreatment GS (2.2, 1.5-3.2), prostate-specific antigen (PSA) level (2.1, 1.1-4.2), perineural invasion (PNI) (1.6, 1.2-2.2), and clinical T-stage (cT) (1.9, 1.0-3.7). • Combination of the independent markers (PSA level > 20 ng/mL or GS >3 + 4 or PNI >3 or cT >2) yielded best risk stratification (RR 11.6, 10.4-12.7). • GS remains one of the most important prognostic factors in prostate cancer. However, the refined guidelines by ISUP 2005 might have shifted the threshold between low-grade and high-grade cancers from GS 6 vs 7 to GS 3 + 4 vs 4 + 3. • PNI is an independent prognostic marker superior to cT. • Ki-67 is the most useful biomarker in detecting patients with GS = 7 at high risk for progression. © 2011 THE AUTHORS. BJU INTERNATIONAL

  3. Effects of Low-Fat Diets Differing in Protein and Carbohydrate Content on Cardiometabolic Risk Factors during Weight Loss and Weight Maintenance in Obese Adults with Type 2 Diabetes.

    PubMed

    Watson, Nerylee; Dyer, Kathryn; Buckley, Jonathan; Brinkworth, Grant; Coates, Alison; Parfitt, Gaynor; Howe, Peter; Noakes, Manny; Murphy, Karen

    2016-05-12

    Despite evidence for the benefits of higher-protein (HP) diets in weight loss, their role in type 2 diabetes mellitus (T2DM) management and weight maintenance is not clear. This randomised study compared the effects of a HP diet (38% carbohydrate, 30% protein, 29% fat) to a isocaloric higher-carbohydrate diet (HC: 53%:21%:23%) on cardiometabolic risk factors for 12 weeks in energy restriction (~30% reduction) followed by 12 weeks of energy balance whilst performing regular exercise. Outcomes were measured at baseline and the end of each phase. Sixty-one overweight/obese adults (BMI (body mass index) 34.3 ± 5.1 kg/m², aged 55 ± 8 years) with T2DM who commenced the study were included in the intention-to-treat analysis including the 17 participants (HP n = 9, HC n = 8) who withdrew. Following weight loss (M ± SEM: -7.8 ± 0.6 kg), there were significant reductions in HbA1c (-1.4% ± 0.1%, p < 0.001) and several cardiometabolic health risk factors. Improvements were sustained for 12 weeks when weight was stabilised and weight loss maintained. Both the HP and HC dietary patterns with concurrent exercise may be effective strategies for weight loss and weight maintenance in T2DM although further studies are needed to determine the longer term effects of weight maintenance.

  4. Grounds Maintenance Cost Report.

    ERIC Educational Resources Information Center

    Joice, Donald K.

    A grounds maintenance study was accomplished over the period of March 1965 through February 1966 to--(1) determine current grounds maintenance cost distribution; (2) provide basis for future grounds maintenance budgeting, and (3) provide basic data by which future landscape project designs can be developed to project or reduce maintenance costs.…

  5. Cryogenics maintenance strategy

    NASA Astrophysics Data System (ADS)

    Cruzat, Fabiola

    2012-09-01

    ALMA is an interferometer composed of 66 independent systems, with specific maintenance requirements for each subsystem. To optimize the observation time and reduce downtime maintenance, requirements are very demanding. One subsystem with high maintenance efforts is cryogenics and vacuum. To organize the maintenance, the Cryogenic and Vacuum department is using and implementing different tools. These are monitoring and problem reporting systems and CMMS. This leads to different maintenance approaches: Preventive Maintenance, Corrective Maintenance and Condition Based Maintenance. In order to coordinate activities with other departments the preventive maintenance schedule is kept as flexible as systems allow. To cope with unavoidable failures, the team has to be prepared to work under any condition with the spares on time. Computerized maintenance management system (CMMS) will help to manage inventory control for reliable spare part handling, the correct record of work orders and traceability of maintenance activities. For an optimized approach the department is currently evaluating where preventive or condition based maintenance applies to comply with the individual system demand. Considering the change from maintenance contracts to in-house maintenance will help to minimize costs and increase availability of parts. Due to increased number of system and tasks the cryo team needs to grow. Training of all staff members is mandatory, in depth knowledge must be built up by doing complex maintenance activities in the Cryo group, use of advanced computerized metrology systems.

  6. Establishing maintenance performance indicators

    SciTech Connect

    Baca, B.

    1994-10-01

    Maintenance Performance Indicators (PI) specify where the maintenance department is and which direction it is going allowing for a quick and accurate assessment of the performance of the Maintenance Management Program (MMP). Establishing PI`s for the maintenance department will allow a measure of productivity and a means of feedback for methods improvement. Effective performance of the maintenance department directly effects plant profitability. Improvements in the quality and productivity of the maintenance work force will significantly reduce maintenance costs. The level of performance attained by the maintenance work force is usually guessed at. Guessing will not identify areas needing improvement or help to initiate a corrective action. Maintenance PI`s are required for maintenance departments whose goal is to control maintenance costs while increasing productivity. The application of basic statistical methods will allow a maintenance department to know where they are and which direction they are going. The data presented in this paper is a representation of indicators used in industry as well as developed indicators to establish a complete maintenance performance indicator program. The methodology used in developing this program can be used as a way to manage a cost effective maintenance management program.

  7. Facilities maintenance handbook

    NASA Technical Reports Server (NTRS)

    1991-01-01

    This handbook is a guide for facilities maintenance managers. Its objective is to set minimum facilities maintenance standards. It also provides recommendations on how to meet the standards to ensure that NASA maintains its facilities in a manner that protects and preserves its investment in the facilities in a cost-effective manner while safely and efficiently performing its mission. This handbook implements NMI 8831.1, which states NASA facilities maintenance policy and assigns organizational responsibilities for the management of facilities maintenance activities on all properties under NASA jurisdiction. It is a reference for facilities maintenance managers, not a step-by-step procedural manual. Because of the differences in NASA Field Installation organizations, this handbook does not assume or recommend a typical facilities maintenance organization. Instead, it uses a systems approach to describe the functions that should be included in any facilities maintenance management system, regardless of its organizational structure. For documents referenced in the handbook, the most recent version of the documents is applicable. This handbook is divided into three parts: Part 1 specifies common definitions and facilities maintenance requirements and amplifies the policy requirements contained in NMI 8831. 1; Part 2 provides guidance on how to meet the requirements of Part 1, containing recommendations only; Part 3 contains general facilities maintenance information. One objective of this handbook is to fix commonality of facilities maintenance definitions among the Centers. This will permit the application of uniform measures of facilities conditions, of the relationship between current replacement value and maintenance resources required, and of the backlog of deferred facilities maintenance. The utilization of facilities maintenance system functions will allow the Centers to quantitatively define maintenance objectives in common terms, prepare work plans, and

  8. Maintenance Management System

    DTIC Science & Technology

    1990-06-01

    maintenance opera- tions. d. Available national maintenance management system (MMS) software be utilized to develop the planning, organizing...portland cement concrete pavements to level and realign faulted areas between slabs or craks within the slab by grinding the high side. MAINTENANCE ITEM

  9. Asphalt in Pavement Maintenance.

    ERIC Educational Resources Information Center

    Asphalt Inst., College Park, MD.

    Maintenance methods that can be used equally well in all regions of the country have been developed for the use of asphalt in pavement maintenance. Specific information covering methods, equipment and terminology that applies to the use of asphalt in the maintenance of all types of pavement structures, including shoulders, is provided. In many…

  10. Defer Maintenance, Invite Disaster

    ERIC Educational Resources Information Center

    Bowman, William W.

    1977-01-01

    An AGB- and NACUBO-sponsored survey showed that "wish lists" are accumulating overdue major maintenance projects because energy costs are consuming physical plant budgets. Problem areas are discussed: budget "guesstimation," preventive maintenance, deferred maintenance inventory, the APPA accounting format, resource allocation,…

  11. Microform Reader Maintenance.

    ERIC Educational Resources Information Center

    Hall, Hal W.; Michaels, George H.

    1985-01-01

    Describes experiences in organizing a program of microform reader and reader/printer maintenance at Texas A & M's Sterling C. Evans Library and offers guidelines for regular machine maintenance and repair. Guidelines discussed relate to maintenance philosophy, general machine cleaning, troubleshooting, service contracts, supplies,…

  12. Light Vehicle Preventive Maintenance.

    ERIC Educational Resources Information Center

    Marine Corps Inst., Washington, DC.

    This correspondence course, originally developed for the Marine Corps, is designed to instruct students in the performance of preventive maintenance on motor vehicles. Instructional materials are presented in three chapters as follows: (1) Major Maintenance Areas (maintenance system, tires, batteries, cooling systems, and vehicle lubrication; (2)…

  13. The ortholog of human solute carrier family 35 member B1 (UDP-galactose transporter-related protein 1) is involved in maintenance of ER homeostasis and essential for larval development in Caenorhabditis elegans.

    PubMed

    Dejima, Katsufumi; Murata, Daisuke; Mizuguchi, Souhei; Nomura, Kazuko H; Gengyo-Ando, Keiko; Mitani, Shohei; Kamiyama, Shin; Nishihara, Shoko; Nomura, Kazuya

    2009-07-01

    Although the solute carrier 35B1 (SLC35B1) is evolutionarily conserved, its functions in metazoans remain unknown. To elucidate its function, we examined developmental roles of an SLC35B1 family gene (HUT-1: homolog of UDP-Gal transporter) in Caenorhabditis elegans. We isolated a deletion mutant of the gene and characterized phenotypes of the mutant and hut-1 RNAi-treated worms. GFP-HUT-1 reporter analysis was performed to examine gene expression patterns. We also tested whether several nucleotide sugar transporters can compensate for hut-1 deficiency. The hut-1 deletion mutant and RNAi worms showed larval growth defect and lethality with disrupted intestinal morphology. Inactivation of hut-1 induced chronic endoplasmic reticulum (ER) stress, and hut-1 showed genetic interactions with the atf-6, pek-1, and ire-1 genes involved in unfolded protein response signaling. ER ultrastructure and ER marker distribution in hut-1-deficient animals showed that HUT-1 is required for maintenance of ER structure. Reporter analysis revealed that HUT-1 is an ER protein ubiquitously expressed in tissues, including the intestine. Lethality and the ER stress phenotype of the mutant were rescued with the human hut-1 ortholog UGTrel1. These results indicate important roles for hut-1 in development and maintenance of ER homeostasis in C. elegans.

  14. Innovation for maintenance technology improvements

    NASA Technical Reports Server (NTRS)

    Shives, T. R. (Editor); Willard, W. A. (Editor)

    1982-01-01

    A group of 34 submitted entries (32 papers and 2 abstracts) from the 33rd meeting of the Mechanical Failures Prevention Group whose subject was maintenance technology improvement through innovation. Areas of special emphasis included maintenance concepts, maintenance analysis systems, improved maintenance processes, innovative maintenance diagnostics and maintenance indicators, and technology improvements for power plant applications.

  15. The actin-binding protein profilin is required for germline stem cell maintenance and germ cell enclosure by somatic cyst cells.

    PubMed

    Shields, Alicia R; Spence, Allyson C; Yamashita, Yukiko M; Davies, Erin L; Fuller, Margaret T

    2014-01-01

    Specialized microenvironments, or niches, provide signaling cues that regulate stem cell behavior. In the Drosophila testis, the JAK-STAT signaling pathway regulates germline stem cell (GSC) attachment to the apical hub and somatic cyst stem cell (CySC) identity. Here, we demonstrate that chickadee, the Drosophila gene that encodes profilin, is required cell autonomously to maintain GSCs, possibly facilitating localization or maintenance of E-cadherin to the GSC-hub cell interface. Germline specific overexpression of Adenomatous Polyposis Coli 2 (APC2) rescued GSC loss in chic hypomorphs, suggesting an additive role of APC2 and F-actin in maintaining the adherens junctions that anchor GSCs to the niche. In addition, loss of chic function in the soma resulted in failure of somatic cyst cells to maintain germ cell enclosure and overproliferation of transit-amplifying spermatogonia.

  16. The actin-binding protein profilin is required for germline stem cell maintenance and germ cell enclosure by somatic cyst cells

    PubMed Central

    Shields, Alicia R.; Spence, Allyson C.; Yamashita, Yukiko M.; Davies, Erin L.; Fuller, Margaret T.

    2014-01-01

    Specialized microenvironments, or niches, provide signaling cues that regulate stem cell behavior. In the Drosophila testis, the JAK-STAT signaling pathway regulates germline stem cell (GSC) attachment to the apical hub and somatic cyst stem cell (CySC) identity. Here, we demonstrate that chickadee, the Drosophila gene that encodes profilin, is required cell autonomously to maintain GSCs, possibly facilitating localization or maintenance of E-cadherin to the GSC-hub cell interface. Germline specific overexpression of Adenomatous Polyposis Coli 2 (APC2) rescued GSC loss in chic hypomorphs, suggesting an additive role of APC2 and F-actin in maintaining the adherens junctions that anchor GSCs to the niche. In addition, loss of chic function in the soma resulted in failure of somatic cyst cells to maintain germ cell enclosure and overproliferation of transit-amplifying spermatogonia. PMID:24346697

  17. [Maintenance of the patterns of expression of homeotic genes in the development of Drosophila melanogaster by proteins of the polycomb, trithorax, and ETP groups].

    PubMed

    Fedorova, E V; Pindiurin, A V; Baricheva, E M

    2009-10-01

    Proteins encoded by genes of the groups Polycomb (PcG), trithorax (trxG), and the Enhancer of Trithorax and Polycomb Group (ETP) are important regulators of expression of most developmental genes. Data concerning all currently described genes assigned to these groups are summarized in the review. Genetic interactions of these genes and phenotypical manifestation of their mutations are described. Data on the PcG, trxG, and ETP proteins are systemized. Questions are considered concerning the formation of multimeric complexes containing proteins of these groups, recruitment of these complexes to regulatory elements of target genes, and the mechanisms of activation/repression of gene expression.

  18. A Potential Role for the Interaction of Wolbachia Surface Proteins with the Brugia malayi Glycolytic Enzymes and Cytoskeleton in Maintenance of Endosymbiosis

    PubMed Central

    Melnikow, Elena; Xu, Shulin; Liu, Jing; Bell, Aaron J.; Ghedin, Elodie; Unnasch, Thomas R.; Lustigman, Sara

    2013-01-01

    The human filarial parasite Brugia malayi harbors an endosymbiotic bacterium of the genus Wolbachia. The Wolbachia represent an attractive target for the control of filarial induced disease as elimination of the bacteria affects molting, reproduction and survival of the worms. The molecular basis for the symbiotic relationship between Wolbachia and their filarial hosts has yet to be elucidated. To identify proteins involved in this process, we focused on the Wolbachia surface proteins (WSPs), which are known to be involved in bacteria-host interactions in other bacterial systems. Two WSP-like proteins (wBm0152 and wBm0432) were localized to various host tissues of the B. malayi female adult worms and are present in the excretory/secretory products of the worms. We provide evidence that both of these proteins bind specifically to B. malayi crude protein extracts and to individual filarial proteins to create functional complexes. The wBm0432 interacts with several key enzymes involved in the host glycolytic pathway, including aldolase and enolase. The wBm0152 interacts with the host cytoskeletal proteins actin and tubulin. We also show these interactions in vitro and have verified that wBm0432 and B. malayi aldolase, as well as wBm0152 and B. malayi actin, co-localize to the vacuole surrounding Wolbachia. We propose that both WSP protein complexes interact with each other via the aldolase-actin link and/or via the possible interaction between the host's enolase and the cytoskeleton, and play a role in Wolbachia distribution during worm growth and embryogenesis. PMID:23593519

  19. A potential role for the interaction of Wolbachia surface proteins with the Brugia malayi glycolytic enzymes and cytoskeleton in maintenance of endosymbiosis.

    PubMed

    Melnikow, Elena; Xu, Shulin; Liu, Jing; Bell, Aaron J; Ghedin, Elodie; Unnasch, Thomas R; Lustigman, Sara

    2013-01-01

    The human filarial parasite Brugia malayi harbors an endosymbiotic bacterium of the genus Wolbachia. The Wolbachia represent an attractive target for the control of filarial induced disease as elimination of the bacteria affects molting, reproduction and survival of the worms. The molecular basis for the symbiotic relationship between Wolbachia and their filarial hosts has yet to be elucidated. To identify proteins involved in this process, we focused on the Wolbachia surface proteins (WSPs), which are known to be involved in bacteria-host interactions in other bacterial systems. Two WSP-like proteins (wBm0152 and wBm0432) were localized to various host tissues of the B. malayi female adult worms and are present in the excretory/secretory products of the worms. We provide evidence that both of these proteins bind specifically to B. malayi crude protein extracts and to individual filarial proteins to create functional complexes. The wBm0432 interacts with several key enzymes involved in the host glycolytic pathway, including aldolase and enolase. The wBm0152 interacts with the host cytoskeletal proteins actin and tubulin. We also show these interactions in vitro and have verified that wBm0432 and B. malayi aldolase, as well as wBm0152 and B. malayi actin, co-localize to the vacuole surrounding Wolbachia. We propose that both WSP protein complexes interact with each other via the aldolase-actin link and/or via the possible interaction between the host's enolase and the cytoskeleton, and play a role in Wolbachia distribution during worm growth and embryogenesis.

  20. Nonredundant roles of mitochondria-associated F-box proteins Mfb1 and Mdm30 in maintenance of mitochondrial morphology in yeast.

    PubMed

    Dürr, Mark; Escobar-Henriques, Mafalda; Merz, Sandra; Geimer, Stefan; Langer, Thomas; Westermann, Benedikt

    2006-09-01

    Mitochondria constantly fuse and divide to adapt organellar morphology to the cell's ever-changing physiological conditions. Little is known about the molecular mechanisms regulating mitochondrial dynamics. F-box proteins are subunits of both Skp1-Cullin-F-box (SCF) ubiquitin ligases and non-SCF complexes that regulate a large number of cellular processes. Here, we analyzed the roles of two yeast F-box proteins, Mfb1 and Mdm30, in mitochondrial dynamics. Mfb1 is a novel mitochondria-associated F-box protein. Mitochondria in mutants lacking Mfb1 are fusion competent, but they form aberrant aggregates of interconnected tubules. In contrast, mitochondria in mutants lacking Mdm30 are highly fragmented due to a defect in mitochondrial fusion. Fragmented mitochondria are docked but nonfused in Deltamdm30 cells. Mitochondrial fusion is also blocked during sporulation of homozygous diploid mutants lacking Mdm30, leading to a mitochondrial inheritance defect in ascospores. Mfb1 and Mdm30 exert nonredundant functions and likely have different target proteins. Because defects in F-box protein mutants could not be mimicked by depletion of SCF complex and proteasome core subunits, additional yet unknown factors are likely involved in regulating mitochondrial dynamics. We propose that mitochondria-associated F-box proteins Mfb1 and Mdm30 are key components of a complex machinery that regulates mitochondrial dynamics throughout yeast's entire life cycle.

  1. Industrial Maintenance Strategies

    SciTech Connect

    Sajjad Akbar

    2006-07-01

    Industrial plants have become more complex due to technological advancement. This has made the task of maintenance more difficult. The maintenance costs in terms of resources and downtime loss are so high that maintenance function has become a critical factor in a plant's profitability. Industry should devote as much forethought to the management of maintenance function as to production. Maintenance has grown from an art to a precise, technical engineering science. Planning, organizing scheduling and control of maintenance using modern techniques pays dividends in the form of reduced costs and increased reliability. The magnitude and the dimension of maintenance have multiplied due to development in the engineering technologies. Production cost and capacities are directly affected by the breakdown time. Total operating cost including the maintenance cost plays an important role in replacement dimension. The integrated system approach would bring forth the desired results of high maintenance standards. The standards once achieved and sustained, would add to the reliability of the plan and relieve heavy stresses and strains on the engineering logistic support. (author)

  2. Examining maintenance responsibilities.

    PubMed

    Lam, K C

    2001-06-01

    This paper has examined the important responsibilities of the two organisations involved in the provision of maintenance service for the vital building services in many of our highly serviced buildings. The issues raised could be put to beneficial use in both clients and maintenance providers. All in all, the clients should work closely with their maintenance providers. Engineering services in buildings will not perform satisfactorily and efficiently if both parties do not work together and understand the maintenance tasks based on a business partnering mode. Put forward is the view that the management of the activities involved in the operation and maintenance process is a "shared commitment/involvement" between the client and the maintenance provider. It is obvious that many factors can influence the continued effectiveness of a quality maintenance scheme set up by client and provider. Some of these factors are: Change in key personnel Updates in technology Amendments to engineering practice Implementation of legislative requirements Changes in operation by client or provider Change of use of building Passage of time These factors must be fully reviewed by both parties from time to time, and necessary actions taken. A cooperative team working relationship and improved communication should be fostered by the client and his provider for the best management of services maintenance. This arrangement will contribute to better building services systems with continuous improvement; improved value for clients and higher return for the maintenance provider.

  3. [Medical Equipment Maintenance Methods].

    PubMed

    Liu, Hongbin

    2015-09-01

    Due to the high technology and the complexity of medical equipment, as well as to the safety and effectiveness, it determines the high requirements of the medical equipment maintenance work. This paper introduces some basic methods of medical instrument maintenance, including fault tree analysis, node method and exclusive method which are the three important methods in the medical equipment maintenance, through using these three methods for the instruments that have circuit drawings, hardware breakdown maintenance can be done easily. And this paper introduces the processing methods of some special fault conditions, in order to reduce little detours in meeting the same problems. Learning is very important for stuff just engaged in this area.

  4. Analysis of mutations in Neurospora crassa ERMES components reveals specific functions related to β-barrel protein assembly and maintenance of mitochondrial morphology.

    PubMed

    Wideman, Jeremy G; Lackey, Sebastian W K; Srayko, Martin A; Norton, Kacie A; Nargang, Frank E

    2013-01-01

    The endoplasmic reticulum mitochondria encounter structure (ERMES) tethers the er to mitochondria and contains four structural components: Mmm1, Mdm12, Mdm10, and Mmm2 (Mdm34). The Gem1 protein may play a role in regulating ERMES function. Saccharomyces cerevisiae and Neurospora crassa strains lacking any of Mmm1, Mdm12, or Mdm10 are known to show a variety of phenotypic defects including altered mitochondrial morphology and defects in the assembly of β-barrel proteins into the mitochondrial outer membrane. Here we examine ERMES complex components in N. crassa and show that Mmm1 is an ER membrane protein containing a Cys residue near its N-terminus that is conserved in the class Sordariomycetes. The residue occurs in the ER-lumen domain of the protein and is involved in the formation of disulphide bonds that give rise to Mmm1 dimers. Dimer formation is required for efficient assembly of Tom40 into the TOM complex. However, no effects are seen on porin assembly or mitochondrial morphology. This demonstrates a specificity of function and suggests a direct role for Mmm1 in Tom40 assembly. Mutation of a highly conserved region in the cytosolic domain of Mmm1 results in moderate defects in Tom40 and porin assembly, as well as a slight morphological phenotype. Previous reports have not examined the role of Mmm2 with respect to mitochondrial protein import and assembly. Here we show that absence of Mmm2 affects assembly of β-barrel proteins and that lack of any ERMES structural component results in defects in Tom22 assembly. Loss of N. crassa Gem1 has no effect on the assembly of these proteins but does affect mitochondrial morphology.

  5. Analysis of Mutations in Neurospora crassa ERMES Components Reveals Specific Functions Related to β-Barrel Protein Assembly and Maintenance of Mitochondrial Morphology

    PubMed Central

    Wideman, Jeremy G.; Lackey, Sebastian W. K.; Srayko, Martin A.; Norton, Kacie A.; Nargang, Frank E.

    2013-01-01

    The endoplasmic reticulum mitochondria encounter structure (ERMES) tethers the ER to mitochondria and contains four structural components: Mmm1, Mdm12, Mdm10, and Mmm2 (Mdm34). The Gem1 protein may play a role in regulating ERMES function. Saccharomyces cerevisiae and Neurospora crassa strains lacking any of Mmm1, Mdm12, or Mdm10 are known to show a variety of phenotypic defects including altered mitochondrial morphology and defects in the assembly of β-barrel proteins into the mitochondrial outer membrane. Here we examine ERMES complex components in N. crassa and show that Mmm1 is an ER membrane protein containing a Cys residue near its N-terminus that is conserved in the class Sordariomycetes. The residue occurs in the ER-lumen domain of the protein and is involved in the formation of disulphide bonds that give rise to Mmm1 dimers. Dimer formation is required for efficient assembly of Tom40 into the TOM complex. However, no effects are seen on porin assembly or mitochondrial morphology. This demonstrates a specificity of function and suggests a direct role for Mmm1 in Tom40 assembly. Mutation of a highly conserved region in the cytosolic domain of Mmm1 results in moderate defects in Tom40 and porin assembly, as well as a slight morphological phenotype. Previous reports have not examined the role of Mmm2 with respect to mitochondrial protein import and assembly. Here we show that absence of Mmm2 affects assembly of β-barrel proteins and that lack of any ERMES structural component results in defects in Tom22 assembly. Loss of N. crassa Gem1 has no effect on the assembly of these proteins but does affect mitochondrial morphology. PMID:23940790

  6. A diet high in protein, dairy, and calcium attenuates bone loss over twelve months of weight loss and maintenance relative to a conventional high-carbohydrate diet in adults.

    PubMed

    Thorpe, Matthew P; Jacobson, Edward H; Layman, Donald K; He, Xuming; Kris-Etherton, Penny M; Evans, Ellen M

    2008-06-01

    Weight loss causes bone mineral loss. Higher protein diets continue to be criticized for further potential harmful bone effects, including elevated urinary calcium, but may promote bone health if protein sources include dairy. Overweight middle-aged subjects (n = 130, 59 males) were randomized to a diet providing 1.4 g.kg(-1).d(-1) protein and 3 daily servings of dairy (PRO) or 0.8 g.kg(-1).d(-1) protein and 2 daily servings of dairy (CARB) for 4 mo of weight loss plus 8 mo of weight maintenance. Diets prescribed 6276 kJ/d for females and 7113 kJ/d for males. Bone mineral content and density (BMD) for whole body (WB), lumbar spine (LS) and total hip (TH) were measured using dual X-ray absorptiometry, and dietary intake using 3-d weighed food records. Urinary calcium was measured using 24-h collection at 0 and 8 mo for a subsample (n = 42). Participants lost body weight (mean, 95% CI) of 8.2% (7.5-8.9%) at 4 mo, 10.6% (9.5-11.8%) at 8 mo, and 10.5% (8.9-12.0%) at 12 mo without differences between groups at any time (P = 0.64). At 12 mo, PRO BMD was higher by 1.6% (0.3-3.0%) at WB, 2.1% (0.6-3.7%) at LS, and 1.4% (0.2-2.5%) at TH compared with CARB. PRO calcium intake was higher (PRO: 1140 +/- 58 mg/d, CARB: 766 +/- 46; P < 0.01), as was urinary calcium (PRO: 163 +/- 15 mg/d, CARB: 100 +/- 9.2; P < 0.01). A reduced-energy diet supplying 1.4 g.kg(-1).d(-1) protein and 3 dairy servings increased urinary calcium excretion but provided improved calcium intake and attenuated bone loss over 4 mo of weight loss and 8 additional mo of weight maintenance.

  7. Mitochondrial fission proteins Fis1 and Mdv1, but not Dnm1, play a role in maintenance of heteroplasmy in budding yeast.

    PubMed

    Bradshaw, Elliot; Yoshida, Minoru; Ling, Feng

    2012-04-24

    In budding yeast, the mitochondrial DNA (mtDNA) replication pathway involving the homologous DNA pairing protein Mhr1 promotes mitochondrial allele segregation. Mitochondrial fusion facilitates the recombination-mediated replication pathway; however, the role of fission remains largely unknown. By monitoring mitochondrial allele segregation during zygotic division, we found that the absence of fission proteins Fis1 or Mdv1, but not Dnm1, resulted in increased initial homoplasmy levels and decreased mtDNA copy number. However, decreases in mtDNA copy number alone were not sufficient for rapid establishment of homoplasmy, suggesting that inhibiting the activities of certain fission proteins promotes homoplasmy by reducing the number of mtDNA segregation units. Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  8. Transcriptional responses indicate maintenance of photosynthetic proteins as key to the exceptional chilling tolerance of C4 photosynthesis in Miscanthus × giganteus

    PubMed Central

    Spence, Ashley K.; Boddu, Jay; Wang, Dafu; James, Brandon; Swaminathan, Kankshita; Moose, Stephen P.; Long, Stephen P.

    2014-01-01

    Miscanthus × giganteus is exceptional among C4 plants in its ability to acclimate to chilling (≤14 °C) and maintain a high photosynthetic capacity, in sharp contrast to maize, leading to very high productivity even in cool temperate climates. To identify the mechanisms that underlie this acclimation, RNA was isolated from M × giganteus leaves in chilling and nonchilling conditions and hybridized to microarrays developed for its close relative Zea mays. Among 21 000 array probes that yielded robust signals, 723 showed significant expression change under chilling. Approximately half of these were for annotated genes. Thirty genes associated with chloroplast membrane function were all upregulated. Increases in transcripts for the lhcb5 (chlorophyll a/b-binding protein CP26), ndhF (NADH dehydrogenase F, chloroplast), atpA (ATP synthase alpha subunit), psbA (D1), petA (cytochrome f), and lhcb4 (chlorophyll a/b-binding protein CP29), relative to housekeeping genes in M. × giganteus, were confirmed by quantitative reverse-transcription PCR. In contrast, psbo1, lhcb5, psbA, and lhcb4 were all significantly decreased in Z. mays after 14 days of chilling. Western blot analysis of the D1 protein and LHCII type II chlorophyll a/b-binding protein also showed significant increases in M. × giganteus during chilling and significant decreases in Z. mays. Compared to other C4 species, M. × giganteus grown in chilling conditions appears to counteract the loss of photosynthetic proteins and proteins protecting photosystem II typically observed in other species by increasing mRNA levels for their synthesis. PMID:24958895

  9. KRE genes are required for beta-1,6-glucan synthesis, maintenance of capsule architecture and cell wall protein anchoring in Cryptococcus neoformans.

    PubMed

    Gilbert, Nicole M; Donlin, Maureen J; Gerik, Kimberly J; Specht, Charles A; Djordjevic, Julianne T; Wilson, Christabel F; Sorrell, Tania C; Lodge, Jennifer K

    2010-04-01

    The polysaccharide beta-1,6-glucan is a major component of the cell wall of Cryptococcus neoformans, but its function has not been investigated in this fungal pathogen. We have identified and characterized seven genes, belonging to the KRE family, which are putatively involved in beta-1,6-glucan synthesis. The H99 deletion mutants kre5Delta and kre6Deltaskn1Delta contained less cell wall beta-1,6-glucan, grew slowly with an aberrant morphology, were highly sensitive to environmental and chemical stress and were avirulent in a mouse inhalation model of infection. These two mutants displayed alterations in cell wall chitosan and the exopolysaccharide capsule, a primary cryptococcal virulence determinant. The cell wall content of the GPI-anchored phospholipase B1 (Plb1) enzyme, which is required for cryptococcal cell wall integrity and virulence, was reduced in kre5Delta and kre6Deltaskn1Delta. Our results indicate that KRE5, KRE6 and SKN1 are involved in beta-1,6-glucan synthesis, maintenance of cell wall integrity and retention of mannoproteins and known cryptococcal virulence factors in the cell wall of C. neoformans. This study sets the stage for future investigations into the function of this abundant cell wall polymer.

  10. KRE genes are required for β-1,6-glucan synthesis, maintenance of capsule architecture and cell wall protein anchoring in Cryptococcus neoformans

    PubMed Central

    Gilbert, Nicole M.; Donlin, Maureen J.; Gerik, Kimberly J.; Specht, Charles A.; Djordjevic, Julianne T.; Wilson, Christabel F.; Sorrell, Tania C.; Lodge, Jennifer K.

    2010-01-01

    Summary The polysaccharide β-1,6-glucan is a major component of the cell wall of Cryptococcus neoformans, but its function has not been investigated in this fungal pathogen. We have identified and characterized seven genes, belonging to the KRE family, which are putatively involved in β-1,6-glucan synthesis. The H99 deletion mutants kre5Δ and kre6Δskn1Δ contained less cell wall β-1,6-glucan, grew slowly with an aberrant morphology, were highly sensitive to environmental and chemical stress and were avirulent in a mouse inhalation model of infection. These two mutants displayed alterations in cell wall chitosan and the exopolysaccharide capsule, a primary cryptococcal virulence determinant. The cell wall content of the GPI-anchored phospholipase B1 (Plb1) enzyme, which is required for cryptococcal cell wall integrity and virulence, was reduced in kre5Δ and kre6Δskn1Δ. Our results indicate that KRE5, KRE6 and SKN1 are involved in β-1,6-glucan synthesis, maintenance of cell wall integrity and retention of mannoproteins and known cryptococcal virulence factors in the cell wall of C. neoformans. This study sets the stage for future investigations into the function of this abundant cell wall polymer. PMID:20384682

  11. Tumor suppressor protein DAB2IP participates in chromosomal stability maintenance through activating spindle assembly checkpoint and stabilizing kinetochore-microtubule attachments

    PubMed Central

    Yu, Lan; Shang, Zeng-Fu; Abdisalaam, Salim; Lee, Kyung-Jong; Gupta, Arun; Hsieh, Jer-Tsong; Asaithamby, Aroumougame; Chen, Benjamin P.C.; Saha, Debabrata

    2016-01-01

    Defects in kinetochore-microtubule (KT-MT) attachment and the spindle assembly checkpoint (SAC) during cell division are strongly associated with chromosomal instability (CIN). CIN has been linked to carcinogenesis, metastasis, poor prognosis and resistance to cancer therapy. We previously reported that the DAB2IP is a tumor suppressor, and that loss of DAB2IP is often detected in advanced prostate cancer (PCa) and is indicative of poor prognosis. Here, we report that the loss of DAB2IP results in impaired KT-MT attachment, compromised SAC and aberrant chromosomal segregation. We discovered that DAB2IP directly interacts with Plk1 and its loss inhibits Plk1 kinase activity, thereby impairing Plk1-mediated BubR1 phosphorylation. Loss of DAB2IP decreases the localization of BubR1 at the kinetochore during mitosis progression. In addition, the reconstitution of DAB2IP enhances the sensitivity of PCa cells to microtubule stabilizing drugs (paclitaxel, docetaxel) and Plk1 inhibitor (BI2536). Our findings demonstrate a novel function of DAB2IP in the maintenance of KT-MT structure and SAC regulation during mitosis which is essential for chromosomal stability. PMID:27568005

  12. Arabidopsis AtNaKR1 is a phloem mobile metal-binding protein necessary for phloem function and root meristem maintenance

    USDA-ARS?s Scientific Manuscript database

    The SODIUM POTASSIUM ROOT DEFECTIVE 1 (NaKR1) encodes a soluble metal binding protein that is specifically expressed in companion cells of the phloem. The nakr1-1 mutant phenotype includes high Na+, K+, and Rb+ accumulation in leaves, short roots, and late flowering. Starch accumulation in the leave...

  13. Differences in basic proline-rich proteins in rat parotid saliva following chronic isoproterenol treatment or maintenance on a liquid diet.

    PubMed

    Johnson, D A

    1983-01-01

    The basic proline-rich proteins (BPRP) in the stimulated parotid saliva of rats treated for 8 days with isoproterenol and rats fed a liquid diet for 2 weeks were compared to those in the stimulated parotid saliva of untreated rats fed a stock pelleted-diet (control). In the control, the BPRP were separated into 5 groups designated Peak A (the basic proline-rich glycoprotein), SP-1, SP-2, SP-3 and SP-4. The percentage of BPRP in each group was as follows: Peak A, 6.5 per cent; SP-1, 37 per cent; SP-2, 6.5 per cent; SP-3, 32.4 per cent; SP-4, 17.6 per cent. In the parotid saliva of rats fed the liquid diet, proteins corresponding to Peak A and SP-2 were not present, the proportion of BPRP in SP-4 was increased to almost 90 per cent while the proportions of material in SP-1 and SP-3 were reduced to 3 and 8 per cent, respectively. In the saliva of rats subjected to chronic isoproterenol treatment, a protein corresponding to SP-4 was not present; proteins corresponding to Peak A, SP-1 and SP-3 were present and in amounts similar to their proportion in untreated rats although material in SP-2 increased to 36 per cent.

  14. HBXIP, a binding protein of HBx, regulates maintenance of the G2/M phase checkpoint induced by DNA damage and enhances sensitivity to doxorubicin-induced cytotoxicity.

    PubMed

    Fei, Hongrong; Zhou, Yunsheng; Li, Ruotong; Yang, Mingfeng; Ma, Jian; Wang, Fengze

    2017-03-04

    To maintain the integrity of the genome, cells need to detect and repair DNA damage before they complete cell division. Hepatitis B x-interacting protein (HBXIP), a binding protein of HBx (Hepatitis B virus × protein), is aberrantly overexpressed in human cancer cells and show to promote cell proliferation and inhibit apoptosis. The present study is designed to investigate the role of HBXIP on the DNA damage response. Our results show that HBXIP acts as an important regulator of G2/M checkpoint in response to DNA damage. HBXIP knockdown increases phospho-histone H2AX expression and foci formation after treatment with ionizing radiation (IR). HBXIP regulates the ATM-Chk2 pathway following DNA damage. Depletion of HBXIP abrogates IR-induced G2/M cell cycle checkpoints, accompanying decrease the expression of phospho-Cdc25C, phospho-Cdc2 (Tyr15) and p27. We also show that downregulation of HBXIP expression sensitizes cancer cells to chemotherapy, as evidenced by an increase in apoptosis and cleavage of caspase-3 and caspase-9. Our data suggest that HBXIP can function as a mediator protein for DNA damage response signals to activate the G2/M checkpoint to maintain genome integrity and prevent cell death.

  15. The intraflagellar transport protein IFT57 is required for cilia maintenance and regulates IFT-particle-kinesin-II dissociation in vertebrate photoreceptors.

    PubMed

    Krock, Bryan L; Perkins, Brian D

    2008-06-01

    Defects in protein transport within vertebrate photoreceptors can result in photoreceptor degeneration. In developing and mature photoreceptors, proteins targeted to the outer segment are transported through the connecting cilium via the process of intraflagellar transport (IFT). In studies of vertebrate IFT, mutations in any component of the IFT particle typically abolish ciliogenesis, suggesting that IFT proteins are equally required for IFT. To determine whether photoreceptor outer segment formation depends equally on individual IFT proteins, we compared the retinal phenotypes of IFT57 and IFT88 mutant zebrafish. IFT88 mutants failed to form outer segments, whereas IFT57 mutants formed short outer segments with reduced amounts of opsin. Our phenotypic analysis revealed that IFT57 is not essential for IFT, but is required for efficient IFT. In co-immunoprecipitation experiments from whole-animal extracts, we determined that kinesin II remained associated with the IFT particle in the absence of IFT57, but IFT20 did not. Additionally, kinesin II did not exhibit ATP-dependent dissociation from the IFT particle in IFT57 mutants. We conclude that IFT20 requires IFT57 to associate with the IFT particle and that IFT57 and/or IFT20 mediate kinesin II dissociation.

  16. Progressive Planned Maintenance.

    ERIC Educational Resources Information Center

    Lewis, Mary Jo; Jacobs, Richard S.

    A planned maintenance system, which was implemented at Washington State University (WSU), uniquely integrates functions of equipment inventory, scheduling, time reporting, project management, materials inventory, and billing. Management now has immediate access to equipment data, maintenance status, and costs. Staff requirements are readily…

  17. Maintenance of School Gymnasiums.

    ERIC Educational Resources Information Center

    Finchum, R. N.

    1965-01-01

    Procedures are suggested that may be helpful to those responsible for the operation and maintenance of school buildings and gymnasiums. Most schools with gymnasiums utilize them for both instructional and sports purposes. Maintenance of the multipurpose gym are discussed under four subject areas--(1) floors, (2) lighting, (3) sanitation, and (4)…

  18. Maintenance of School Gymnasiums.

    ERIC Educational Resources Information Center

    Finchum, R. N.

    1965-01-01

    Procedures are suggested that may be helpful to those responsible for the operation and maintenance of school buildings and gymnasiums. Most schools with gymnasiums utilize them for both instructional and sports purposes. Maintenance of the multipurpose gym are discussed under four subject areas--(1) floors, (2) lighting, (3) sanitation, and (4)…

  19. Halt Runaway Maintenance Costs.

    ERIC Educational Resources Information Center

    Trotter, Andrew

    1988-01-01

    To keep expensive computer software and hardware functioning requires good housekeeping, some protective equipment, ground rules, and supervision. Schools can arrange microcomputer maintenance through service agreements with computer manufacturers, pay-as-you-go service from local computer stores, or setting up their own maintenance and repair…

  20. Fixing Maintenance Productivity.

    ERIC Educational Resources Information Center

    Fickes, Michael

    2003-01-01

    Describes how one university's facility managers use Nextel communications technology in conjunction with a Famis Software maintenance management system to improve the productivity of its maintenance technicians. The system uses a wireless Internet connection to automate the flow of work order information to and from technicians. The key to these…

  1. Fixing Maintenance Productivity.

    ERIC Educational Resources Information Center

    Fickes, Michael

    2003-01-01

    Describes how one university's facility managers use Nextel communications technology in conjunction with a Famis Software maintenance management system to improve the productivity of its maintenance technicians. The system uses a wireless Internet connection to automate the flow of work order information to and from technicians. The key to these…

  2. Maximizing Hard Floor Maintenance.

    ERIC Educational Resources Information Center

    Steger, Michael

    2000-01-01

    Explains the maintenance options available for hardwood flooring that can help ensure long life cycles and provide inviting spaces. Developing a maintenance system, knowing the type of traffic that the floor must endure, using entrance matting, and adhering to manufacturers guidelines are discussed. Daily, monthly or quarterly, and long-term…

  3. [Targeted periodontal maintenance].

    PubMed

    Defleur, P; De Beule, F

    1995-01-01

    The goal of periodontal preventive cares known as "periodontal maintenance", are to maintain or to improve the periodontal situation after initial periodontal therapy. The inconveniences of a linear organisation of the maintenance are multiple. The aim of this work is to search a better way to organise the periodontal preventive cares and to offer a better service to the patients.

  4. Maintenance Trades Guidelines

    ERIC Educational Resources Information Center

    Weidner, Theodore J.

    2008-01-01

    In 2002, APPA published "Maintenance Staffing Guidelines for Educational Facilities," the first building maintenance trades staffing guideline designed to assist educational facilities professionals with their staffing needs. addresses how facilities professionals can determine the appropriate size and mix of their organization. Contents…

  5. Defective chloroplast development inhibits maintenance of normal levels of abscisic acid in a mutant of the Arabidopsis RH3 DEAD-box protein during early post-germination growth.

    PubMed

    Lee, Kwang-Hee; Park, Jiyoung; Williams, Donna S; Xiong, Yuqing; Hwang, Inhwan; Kang, Byung-Ho

    2013-03-01

    The plastid has its own translation system, and its ribosomes are assembled through a complex process in which rRNA precursors are processed and ribosomal proteins are inserted into the rRNA backbone. DEAD-box proteins have been shown to play roles in multiple steps in ribosome biogenesis. To investigate the cellular and physiological roles of an Arabidopsis DEAD-box protein, RH3, we examined its expression and localization and the phenotypes of rh3-4, a T-DNA insertion mutant allele of RH3. The promoter activity of RH3 is strongest in the greening tissues of 3-day and 1-week-old seedlings but reduced afterwards. Cotyledons were pale and seedling growth was retarded in the mutant. The most obvious abnormality in the mutant chloroplasts was their lack of normal ribosomes. Electron tomography analysis indicated that ribosome density in the 3-day-old mutant chloroplasts is only 20% that of wild-type chloroplasts, and the ribosomes in the mutant are smaller. These chloroplast defects in rh3-4 were alleviated in 2-week-old cotyledons and true leaves. Interestingly, rh3-4 seedlings have lower amounts of abscisic acid prior to recovery of their chloroplasts, and were more sensitive to abiotic stresses. Transcriptomic analysis indicated that nuclear genes for chloroplast proteins are down-regulated, and proteins mediating chloroplast-localized steps of abscisic acid biosynthesis are expressed to a lower extent in 1-week-old rh3-4 seedlings. Taken together, these results suggest that conversion of eoplasts into chloroplasts in young seedlings is critical for the seedlings to start carbon fixation as well as for maintenance of abscisic acid levels for responding to environmental challenges. © 2012 The Authors The Plant Journal © 2012 Blackwell Publishing Ltd.

  6. Effect of serum fibroblast growth factor-23, matrix Gla protein and Fetuin-A in predicting osteoporosis in maintenance hemodialysis patients.

    PubMed

    Wu, Qing; Xiao, Dong-Mei; Fan, Wei-Feng; Ye, Xian-Wu; Niu, Jian-Ying; Gu, Yong

    2014-10-01

    This study is aimed at exploring the role of serum fibroblast growth factor-23 (FGF-23), matrix Gla (MGP) and Fetuin-A in the calcium-phosphate metabolism and estimate the value of serum FGF-23, MGP and Fetuin-A levels in predicting osteoporosis in maintenance hemodialysis (MHD) patients. This study included 64 patients who receive hemodialysis in our hospital. The serum FGF-23, MGP and Fetuin-A were analyzed by enzyme-linked immunosorbent assay (ELlSA). Bone mineral density (BMD) at the femoral neck was measured by dual-energy X-ray absorptiometry. The 64 patients (30 males, 34 females, 60.6 ± 11.3 years of age) received an average of 6.88 ± 2.94 years of dialysis. Body mass index (BMI), Kt/V, dialysis vintage, patient age, serum levels of FGF-23, Fetuin-A, bone isoenzyme of alkaline phosphatase (ALP-B), and calcium were different in statistical significance among the three groups of patients in terms of normal bone mass (N = 10), osteopenia (N = 24), or osteoporosis (N = 30). BMI, Kt/V, ALP-B, dialysis vintage and serum Fetuin-A level were identified as independent variables of femoral neck BMD by stepwise multiple regression analysis. The area under ROC curve showed that serum Fetuin-A was useful for identifying osteoporosis in MHD patients. The cutoff value corresponding to the highest Youden's index was serum Fetuin-A ≤ 89 μg/mL, which was defined as the optimal predictor of osteoporosis. Its sensitivity/specificity was 71%/77.8%. The incidence of osteoporosis is high in MHD patients. Serum Fetuin-A level is closely correlated with osteoporosis and it may serve as a predictor of osteoporosis.

  7. A new evidence for the maintenance of the sarcoglycan complex in muscle sarcolemma in spite of the primary absence of delta-SG protein.

    PubMed

    Gouveia, Telma L F; Kossugue, Patrícia M; Paim, Julia F; Zatz, Mayana; Anderson, Louise V B; Nigro, Vincenzo; Vainzof, Mariz

    2007-04-01

    delta-Sarcoglycan (delta-SG) is one of the first proteins of the sarcoglycan complex (SGC) to be expressed during muscle development, and it has been considered fundamental for the assembling and insertion of the SGC in the sarcolemma. Studies using heterologous cell systems and co-precipitation have demonstrated that SGC assembly was dependent on the simultaneous synthesis of all four sarcoglycan proteins. Mutations in any one of sarcoglycan genes, including the common disease causing mutation c.656delC in the delta-SG gene, block complex formation and its insertion in the plasma membrane. Failure in complex assembly in patients with this mutation would be therefore expected. In this study, we provide evidence for the possibility of preservation of part of the SG complex in the sarcolemma, even in the absence of delta-SG. This is based on the study of one mildly affected patient with limb-girdle muscular dystrophy type 2F (LGMD2F) due to the homozygous c.656delC mutation in the delta-SG gene. Protein analysis in his muscle biopsy presented a significant deficiency of only delta-SG with retention of the other three SG proteins in the sarcolemma. RNA expression analysis showed that zeta-SG, a functionally homologous to delta-SG, is not atypically upregulated in his muscle and would not replace the absent delta-SG, retaining the complex alpha-beta-gamma-zeta. The patient started clinical manifestation at age 25, with frequent falls, but he is currently able to walk unassisted at age 42. His clinical course is significantly milder when compared to several other affected patients carrying the same mutation associated with a total deficiency of the four SG proteins in the muscle studied by our group and confirmed in other patients. Therefore, our results add a new in vivo evidence that alpha-, beta-, and gamma-SG proteins can be maintained in the sarcolemma without delta-SG. Additionally, LGMD2F, with retention of the part of the SGC, might be associated to a milder

  8. Pkh1 and Pkh2 Differentially Phosphorylate and Activate Ypk1 and Ykr2 and Define Protein Kinase Modules Required for Maintenance of Cell Wall Integrity

    PubMed Central

    Roelants, Françoise M.; Torrance, Pamela D.; Bezman, Natalie; Thorner, Jeremy

    2002-01-01

    Saccharomyces cerevisiae Pkh1 and Pkh2 are functionally redundant homologs of mammalian protein kinase, phosphoinositide-dependent protein kinase-1. They activate two closely related, functionally redundant enzymes, Ypk1 and Ykr2 (homologs of mammalian protein kinase, serum- and glucocorticoid-inducible protein kinase). We found that Ypk1 has a more prominent role than Ykr2 in mediating their shared essential function. Considerable evidence demonstrated that Pkh1 preferentially activates Ypk1, whereas Pkh2 preferentially activates Ykr2. Loss of Pkh1 (but not Pkh2) reduced Ypk1 activity; conversely, Pkh1 overexpression increased Ypk1 activity more than Pkh2 overexpression. Loss of Pkh2 reduced Ykr2 activity; correspondingly, Pkh2 overexpression increased Ykr2 activity more than Pkh1 overexpression. When overexpressed, a catalytically active C-terminal fragment (kinase domain) of Ypk1 was growth inhibitory; loss of Pkh1 (but not Pkh2) alleviated toxicity. Loss of Pkh2 (but not Pkh1) exacerbated the slow growth phenotype of a ypk1Δ strain. This Pkh1-Ypk1 and Pkh2-Ykr2 dichotomy is not absolute because all double mutants (pkh1Δ ypk1Δ, pkh2Δ ypk1Δ, pkh1Δ ykr2Δ, and pkh2Δ ykr2Δ) were viable. Compartmentation contributes to selectivity because Pkh1 and Ypk1 were located exclusively in the cytosol, whereas Pkh2 and Ykr2 entered the nucleus. At restrictive temperature, ypk1-1ts ykr2Δ cells lysed rapidly, but not in medium containing osmotic support. Dosage and extragenic suppressors were selected. Overexpression of Exg1 (major exoglucanase), or loss of Kex2 (endoprotease involved in Exg1 processing), rescued growth at high temperature. Viability was also maintained by PKC1 overexpression or an activated allele of the downstream protein kinase (BCK1-20). Conversely, absence of Mpk1 (distal mitogen-activated protein kinase of the PKC1 pathway) was lethal in ypk1-1ts ykr2Δ cells. Thus, Pkh1-Ypk1 and Pkh2-Ykr2 function in a novel pathway for cell wall integrity that

  9. Cep164 is a mediator protein required for the maintenance of genomic stability through modulation of MDC1, RPA, and CHK1

    PubMed Central

    Sivasubramaniam, Sudhakar; Sun, Xuemin; Pan, Yen-Ru; Wang, Shaohui; Lee, Eva Y.-H.P.

    2008-01-01

    The activation of the ataxia telangiectasia mutated (ATM) and ATM/Rad3-related (ATR) kinases triggers a diverse cellular response including the initiation of DNA damage-induced cell cycle checkpoints. Mediator of DNA Damage Checkpoint protein, MDC1, and H2AX are chromatin remodeling factors required for the recruitment of DNA repair proteins to the DNA damage sites. We identified a novel mediator protein, Cep164 (KIAA1052), that interacts with both ATR and ATM. Cep164 is phosphorylated upon replication stress, ultraviolet radiation (UV), and ionizing radiation (IR). Ser186 of Cep164 is phosphorylated by ATR/ATM in vitro and in vivo. The phosphorylation of Ser186 is not affected by RPA knockdown but is severely hampered by MDC1 knockdown. siRNA-mediated silencing of Cep164 significantly reduces DNA damage-induced phosphorylation of RPA, H2AX, MDC1, CHK2, and CHK1, but not NBS1. Analyses of Cep164 knockdown cells demonstrate a critical role of Cep164 in G2/M checkpoint and nuclear divisions. These findings reveal that Cep164 is a key player in the DNA damage-activated signaling cascade. PMID:18283122

  10. Normal microRNA Maturation and Germ-Line Stem Cell Maintenance Requires Loquacious, a Double-Stranded RNA-Binding Domain Protein

    PubMed Central

    Förstemann, Klaus; Tomari, Yukihide; Du, Tingting; Vagin, Vasily V; Denli, Ahmet M; Bratu, Diana P; Klattenhoff, Carla; Theurkauf, William E

    2005-01-01

    microRNAs (miRNAs) are single-stranded, 21- to 23-nucleotide cellular RNAs that control the expression of cognate target genes. Primary miRNA (pri-miRNA) transcripts are transformed to mature miRNA by the successive actions of two RNase III endonucleases. Drosha converts pri-miRNA transcripts to precursor miRNA (pre-miRNA); Dicer, in turn, converts pre-miRNA to mature miRNA. Here, we show that normal processing of Drosophila pre-miRNAs by Dicer-1 requires the double-stranded RNA-binding domain (dsRBD) protein Loquacious (Loqs), a homolog of human TRBP, a protein first identified as binding the HIV trans-activator RNA (TAR). Efficient miRNA-directed silencing of a reporter transgene, complete repression of white by a dsRNA trigger, and silencing of the endogenous Stellate locus by Suppressor of Stellate, all require Loqs. In loqs f00791 mutant ovaries, germ-line stem cells are not appropriately maintained. Loqs associates with Dcr-1, the Drosophila RNase III enzyme that processes pre-miRNA into mature miRNA. Thus, every known Drosophila RNase-III endonuclease is paired with a dsRBD protein that facilitates its function in small RNA biogenesis. PMID:15918770

  11. Maintenance Process Strategic Analysis

    NASA Astrophysics Data System (ADS)

    Jasiulewicz-Kaczmarek, M.; Stachowiak, A.

    2016-08-01

    The performance and competitiveness of manufacturing companies is dependent on the availability, reliability and productivity of their production facilities. Low productivity, downtime, and poor machine performance is often linked to inadequate plant maintenance, which in turn can lead to reduced production levels, increasing costs, lost market opportunities, and lower profits. These pressures have given firms worldwide the motivation to explore and embrace proactive maintenance strategies over the traditional reactive firefighting methods. The traditional view of maintenance has shifted into one of an overall view that encompasses Overall Equipment Efficiency, Stakeholders Management and Life Cycle assessment. From practical point of view it requires changes in approach to maintenance represented by managers and changes in actions performed within maintenance area. Managers have to understand that maintenance is not only about repairs and conservations of machines and devices, but also actions striving for more efficient resources management and care for safety and health of employees. The purpose of the work is to present strategic analysis based on SWOT analysis to identify the opportunities and strengths of maintenance process, to benefit from them as much as possible, as well as to identify weaknesses and threats, so that they could be eliminated or minimized.

  12. Powerful drivers for maintenance.

    PubMed

    Mason, Steve

    2013-10-01

    A UPS system is the central building block of a Power Continuity Plan in medical facilities, but such equipment requires careful maintenance to continue fulfilling its vital role in delivering power resilience, and avoid catastrophic downtime, and potentially tens of thousands of pounds in costs to rectify the issues caused by poor maintenance. In our latest technical guidance article, Steve Mason, MD at Bender UK, one of the leading providers of isolated power supplies, theatre control panels, UPS systems, and Steris surgical products, and a turnkey provider of solutions for safe handling of electrical power and advanced provision of critical care products, examines some of the issue surrounding UPS maintenance.

  13. Factors Influencing Army Maintenance

    DTIC Science & Technology

    1989-01-01

    ARI Research Note 89-11 (N 00 Factors Influencing Army Maintenance LOloD Debra C. Evans and J. Thomas Roth Applied Science Associates, Inc. for...1.2.7 .2.7.C.1 11. TITLE (Include Security ClassifIcarIon) Factors Influencing Army Maintenance i2. FERSONAL AuTtiOR(S) Evans, Debra C., and Roth, J...y • ’ Factors and variables that influence maintenance for systems and related manpower, per- sonnel, and training (MPT) characteristics were

  14. General Healthcare Maintenance of IBD

    MedlinePlus

    ... Resources > General Healthcare Maintenance Go Back General Healthcare Maintenance Email Print + Share It is important to continue ... Bowel Diseases. September 2009, 1399-1409 Health Care Maintenance Chart YEAR PPD (Tuberculosis skin test) Vaccinations Hepatitis ...

  15. Nuclear localization of the dystrophin-associated protein α-dystrobrevin through importin α2/β1 is critical for interaction with the nuclear lamina/maintenance of nuclear integrity.

    PubMed

    Aguilar, Areli; Wagstaff, Kylie M; Suárez-Sánchez, Rocío; Zinker, Samuel; Jans, David A; Cisneros, Bulmaro

    2015-05-01

    Although α-dystrobrevin (DB) is assembled into the dystrophin-associated protein complex, which is central to cytoskeletal organization, it has also been found in the nucleus. Here we delineate the nuclear import pathway responsible for nuclear targeting of α-DB for the first time, together with the importance of nuclear α-DB in determining nuclear morphology. We map key residues of the nuclear localization signal of α-DB within the zinc finger domain (ZZ) using various truncated versions of the protein, and site-directed mutagenesis. Pulldown, immunoprecipitation, and AlphaScreen assays showed that the importin (IMP) α2/β1 heterodimer interacts with high affinity with the ZZ domain of α-DB. In vitro nuclear import assays using antibodies to specific importins, as well as in vivo studies using siRNA or a dominant negative importin construct, confirmed the key role of IMPα2/β1 in α-DB nuclear translocation. Knockdown of α-DB expression perturbed cell cycle progression in C2C12 myoblasts, with decreased accumulation of cells in S phase and, significantly, altered localization of lamins A/C, B1, and B2 with accompanying gross nuclear morphology defects. Because α-DB interacts specifically with lamin B1 in vivo and in vitro, nuclear α-DB would appear to play a key role in nuclear shape maintenance through association with the nuclear lamina.

  16. Interleukin 7 up-regulates CD95 protein on CD4+ T cells by affecting mRNA alternative splicing: priming for a synergistic effect on HIV-1 reservoir maintenance.

    PubMed

    Yin, Yue; Zhang, Shaoying; Luo, Haihua; Zhang, Xu; Geng, Guannan; Li, Jun; Guo, Xuemin; Cai, Weiping; Li, Linghua; Liu, Chao; Zhang, Hui

    2015-01-02

    Interleukin-7 (IL-7) has been used as an immunoregulatory and latency-reversing agent in human immunodeficiency virus type 1 (HIV-1) infection. Although IL-7 can restore circulating CD4(+) T cell counts in HIV-1-infected patients, the anti-apoptotic and proliferative effects of IL-7 appear to benefit survival and expansion of HIV-1-latently infected memory CD4(+) T lymphocytes. IL-7 has been shown to elevate CD95 on CD4(+) T cells in HIV-1-infected individuals and prime CD4(+) T lymphocytes to CD95-mediated proliferative or apoptotic signals. Here we observed that through increasing microRNA-124, IL-7 down-regulates the splicing regulator polypyrimidine tract binding protein (PTB), leading to inclusion of the transmembrane domain-encoding exon 6 of CD95 mRNA and, subsequently, elevation of CD95 on memory CD4(+) T cells. Moreover, IL-7 up-regulates cellular FLICE-like inhibitory protein (c-FLIP) and stimulates c-Jun N-terminal kinase (JNK) phosphorylation, which switches CD95 signaling to survival mode in memory CD4(+) T lymphocytes. As a result, co-stimulation through IL-7/IL-7R and FasL/CD95 signal pathways augments IL-7-mediated survival and expansion of HIV-1-latently infected memory CD4(+) T lymphocytes. Collectively, we have demonstrated a novel mechanism for IL-7-mediated maintenance of HIV-1 reservoir.

  17. Insight into centromere-binding properties of ParB proteins: a secondary binding motif is essential for bacterial genome maintenance

    PubMed Central

    Sanchez, Aurore; Rech, Jérôme; Gasc, Cyrielle; Bouet, Jean-Yves

    2013-01-01

    ParB proteins are one of the three essential components of partition systems that actively segregate bacterial chromosomes and plasmids. In binding to centromere sequences, ParB assembles as nucleoprotein structures called partition complexes. These assemblies are the substrates for the partitioning process that ensures DNA molecules are segregated to both sides of the cell. We recently identified the sopC centromere nucleotides required for binding to the ParB homologue of plasmid F, SopB. This analysis also suggested a role in sopC binding for an arginine residue, R219, located outside the helix-turn-helix (HTH) DNA-binding motif previously shown to be the only determinant for sopC-specific binding. Here, we demonstrated that the R219 residue is critical for SopB binding to sopC during partition. Mutating R219 to alanine or lysine abolished partition by preventing partition complex assembly. Thus, specificity of SopB binding relies on two distinct motifs, an HTH and an arginine residue, which define a split DNA-binding domain larger than previously thought. Bioinformatic analysis over a broad range of chromosomal ParBs generalized our findings with the identification of a non-HTH positively charged residue essential for partition and centromere binding, present in a newly identified highly conserved motif. We propose that ParB proteins possess two DNA-binding motifs that form an extended centromere-binding domain, providing high specificity. PMID:23345617

  18. Epidermal Fatty Acid Binding Protein (E-FABP) Is Not Required for the Generation or Maintenance of Effector and Memory T Cells following Infection with Listeria monocytogenes

    PubMed Central

    Li, Bing; Schmidt, Nathan W.

    2016-01-01

    Following activation of naïve T cells there are dynamic changes in the metabolic pathways used by T cells to support both the energetic needs of the cell and the macromolecules required for growth and proliferation. Among other changes, lipid metabolism undergoes dynamic transitions between fatty acid oxidation and fatty acid synthesis as cells progress from naïve to effector and effector to memory T cells. The hydrophobic nature of lipids requires that they be bound to protein chaperones within a cell. Fatty acid binding proteins (FABPs) represent a large class of lipid chaperones, with epidermal FABP (E-FABP) expressed in T cells. The objective of this study was to determine the contribution of E-FABP in antigen-specific T cell responses. Following infection with Listeria monocytogenes, we observed similar clonal expansion, contraction and formation of memory CD8 T cells in WT and E-FABP-/- mice, which also exhibited similar phenotypic and functional characteristics. Analysis of Listeria-specific CD4 T cells also revealed no defect in the expansion, contraction, and formation of memory CD4 T cells in E-FABP-/- mice. These data demonstrate that E-FABP is dispensable for antigen-specific T cell responses following a bacterial infection. PMID:27588422

  19. Hice1, a Novel Microtubule-Associated Protein Required for Maintenance of Spindle Integrity and Chromosomal Stability in Human Cells▿ †

    PubMed Central

    Wu, Guikai; Lin, Yi-Tzu; Wei, Randy; Chen, Yumay; Shan, Zhiyin; Lee, Wen-Hwa

    2008-01-01

    Spindle integrity is critical for efficient mitotic progression and accurate chromosome segregation. Deregulation of spindles often leads to structural and functional aberrations, ultimately promoting segregation errors and aneuploidy, a hallmark of most human cancers. Here we report the characterization of a previously identified human sarcoma antigen (gene located at 19p13.11), Hice1, an evolutionarily nonconserved 46-kDa coiled-coil protein. Hice1 shows distinct cytoplasmic localization and associates with interphase centrosomes and mitotic spindles, preferentially at the spindle pole vicinity. Depletion of Hice1 by RNA interference resulted in abnormal and unstable spindle configurations, mitotic delay at prometaphase and metaphase, and elevated aneuploidy. Conversely, loss of Hice1 had minimal effects on interphase centrosome duplication. We also found that both full-length Hice1 and Hice1-N1, which is composed of 149 amino acids of the N-terminal region, but not the mutant lacking the N-terminal region, exhibited activities of microtubule bundling and stabilization at a near-physiological concentration. Consistently, overexpression of Hice1 rendered microtubule bundles in cells resistant to nocodazole- or cold-treatment-induced depolymerization. These results demonstrate that Hice1 is a novel microtubule-associated protein important for maintaining spindle integrity and chromosomal stability, in part by virtue of its ability to bind, bundle, and stabilize microtubules. PMID:18362163

  20. MCA Inflight Maintenance

    NASA Image and Video Library

    2011-07-06

    ISS028-E-014395 (6 July 2011) --- NASA astronaut Ron Garan, Expedition 28 flight engineer, performs in-flight maintenance on the Mass Constituents Analyzer (MCA) in the Destiny laboratory of the International Space Station.

  1. Woodwind Instrument Maintenance.

    ERIC Educational Resources Information Center

    Sperl, Gary

    1980-01-01

    The author presents a simple maintenance program for woodwind instruments which includes the care of tendon corks, the need for oiling keys, and methods of preventing cracks in woodwind instruments. (KC)

  2. Getting into Motorcycle Maintenance

    ERIC Educational Resources Information Center

    Mosher, Lynn

    1978-01-01

    This article tells how to start a high school course in motorcycle maintenance and includes names and addresses of some motorcycle manufacturers and a list of needed tools, equipment, and materials. (MF)

  3. Automating Preventive Maintenance.

    ERIC Educational Resources Information Center

    Oshier, Michael J.

    1984-01-01

    Describes the following aspects of the State University of New York-Brockport's preventive maintenance computerization project: (1) software selection, (2) project implementation; and (3) problems and benefits of the system. (MCG)

  4. Automated preventive maintenance program

    NASA Technical Reports Server (NTRS)

    Cea, E. J.; Grieger, T. H.

    1971-01-01

    Maintenance program which is concise and inexpensive to operate adapts to almost any system that has a FORTRAN compiler. Program operates on a stored data base with an output consisting of scheduling information and various management reports.

  5. Timpani Repair and Maintenance.

    ERIC Educational Resources Information Center

    Combs, F. Michael

    1980-01-01

    Rather than focusing on specific brands of timpani, these guidelines for repair cover mechanical problems of a general nature: pedals, dents, unclear tone, and squeaking. Preventive maintenance is discussed. (Author/SJL)

  6. Care and Maintenance.

    ERIC Educational Resources Information Center

    Hampton, Carolyn H.; Hampton, Carol D.

    1979-01-01

    The classroom care and maintenance of terrestrial isopods is described. Includes illustrations of isopod external anatomy, a potato trap for collecting isopods, and a constructed habitat for raising isopods. (MA)

  7. Woodwind Instrument Maintenance.

    ERIC Educational Resources Information Center

    Sperl, Gary

    1980-01-01

    The author presents a simple maintenance program for woodwind instruments which includes the care of tendon corks, the need for oiling keys, and methods of preventing cracks in woodwind instruments. (KC)

  8. Carpet Maintenance Guide.

    ERIC Educational Resources Information Center

    Griffin, William R.

    1999-01-01

    Discusses ways to make carpet maintenance in schools easier and effective for keeping carpeted areas in schools attractive and long lasting. Covers cleaning tips for basic spills, ideas for staying on top of stains, and suggestions for eliminating odors. (GR)

  9. Carpet Maintenance Guide.

    ERIC Educational Resources Information Center

    Griffin, William R.

    1999-01-01

    Discusses ways to make carpet maintenance in schools easier and effective for keeping carpeted areas in schools attractive and long lasting. Covers cleaning tips for basic spills, ideas for staying on top of stains, and suggestions for eliminating odors. (GR)

  10. Care and Maintenance.

    ERIC Educational Resources Information Center

    Hampton, Carolyn H.; Hampton, Carol D.

    1979-01-01

    The classroom care and maintenance of terrestrial isopods is described. Includes illustrations of isopod external anatomy, a potato trap for collecting isopods, and a constructed habitat for raising isopods. (MA)

  11. The Deferred Maintenance Dilemma.

    ERIC Educational Resources Information Center

    Medlin, E. Lander

    2003-01-01

    Discusses the hazards of the trend toward accumulated deferred maintenance in higher education and offers advice on tying facilities needs and issues to the core strategies and goals of the institution. (EV)

  12. Barrett during TVIS maintenance

    NASA Image and Video Library

    2009-04-21

    ISS019-E-008767 (21 April 2009) --- Astronaut Michael Barratt, Expedition 19/20 flight engineer, performs in-flight maintenance on the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station.

  13. Barrett during TVIS maintenance

    NASA Image and Video Library

    2009-04-21

    ISS019-E-008750 (21 April 2009) --- Astronaut Michael Barratt, Expedition 19/20 flight engineer, performs in-flight maintenance on the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station.

  14. Barrett during TVIS maintenance

    NASA Image and Video Library

    2009-04-21

    ISS019-E-008764 (21 April 2009) --- Astronaut Michael Barratt, Expedition 19/20 flight engineer, performs in-flight maintenance on the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station.

  15. Barrett during TVIS maintenance

    NASA Image and Video Library

    2009-04-21

    ISS019-E-008752 (21 April 2009) --- Astronaut Michael Barratt, Expedition 19/20 flight engineer, performs in-flight maintenance on the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station.

  16. TVIS Inflight Maintenance

    NASA Image and Video Library

    2011-07-01

    ISS028-E-013757 (1 July 2011) --- Russian cosmonaut Andrey Borisenko, Expedition 28 commander, performs in-flight maintenance on the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station.

  17. SPHERES Maintenance Run

    NASA Image and Video Library

    2016-06-30

    iss048e017435 (6/30/2016) --- Commander Jeff Williams monitors bowling ball-sized internal satellites known as SPHERES (Synchronized Position Hold, Engage, Reorient, Experimental Satellites) during a maintenance run in the Japanese Kibo Laboratory Module.

  18. Maintenance of Dimer Conformation by the Dengue Virus Core Protein α4-α4′ Helix Pair Is Critical for Nucleocapsid Formation and Virus Production

    PubMed Central

    Teoh, Pak-Guan; Huang, Zhi-Shun; Pong, Wen-Li; Chen, Po-Chiang

    2014-01-01

    ABSTRACT The virion of dengue virus (DENV) is composed of a viral envelope covering a nucleocapsid formed by a complex of viral genomic RNA and core protein (CP). DENV CP forms a dimer via the internal α2 and α4 helices of each monomer. Pairing of α2-α2′ creates a continuous hydrophobic surface, while the α4-α4′ helix pair joins the homodimer via side-chain interactions of the inner-edge residues. However, the importance of dimer conformation and the α4 helix of DENV CP in relation to its function are poorly understood. Loss of association between CP and lipid droplets (LDs) due to mutation suggests that the CP hydrophobic surface was not exposed, offering a possible explanation for the absence of dimers. Further assays suggest the connection between CP folding and protein stability. Attenuation of full-length RNA-derived virus production is associated with CP mutation, since no significant defects were detected in virus translation and replication. The in vitro characterization assays further highlighted that the α4-α4′ helix pair conformation is critical in preserving the overall α-helical content, thermostability, and dimer formation ability of CP, features correlated with the efficiency of nucleocapsid formation. Addition of Tween 20 improves in vitro nucleocapsid-like particle formation, suggesting the role of the LD in nucleocapsid formation in vivo. This study provides the first direct link between the α4-α4′ helix pair interaction and the CP dimer conformation that is the basis of CP function, particularly in nucleocapsid formation during virion production. IMPORTANCE Structure-based mutagenesis study of the dengue virus core protein (CP) reveals that the α4-α4′ helix pair is the key to maintaining its dimer conformation, which is the basis of CP function in nucleocapsid formation and virus production. Attenuation of full-length RNA-derived virus production is associated with CP mutation, since no significant defects in virus

  19. The AT-hook DNA binding ability of the Epstein Barr virus EBNA1 protein is necessary for the maintenance of viral genomes in latently infected cells.

    PubMed

    Chakravorty, Adityarup; Sugden, Bill

    2015-10-01

    Epstein Barr Virus (EBV) is a human tumor virus that is causally linked to malignancies such as Burkitt׳s lymphoma, and gastric and nasopharyngeal carcinomas. Tethering of EBV genomes to cellular chromosomes is required for the synthesis and persistence of viral plasmids in tumor cells. However, it is not established how EBV genomes are tethered to cellular chromosomes. We test the hypothesis that the viral protein EBNA1 tethers EBV genomes to chromosomes specifically through its N-terminal AT-hook DNA-binding domains by using a small molecule, netropsin, that has been shown to inhibit the AT-hook DNA-binding of EBNA1 in vitro. We show that netropsin forces the loss of EBV genomes from epithelial and lymphoid cells in an AT-hook dependent manner and that EBV-positive lymphoma cells are significantly more inhibited in their growth by netropsin than are corresponding EBV-negative cells.

  20. Reliability-Centered Maintenance

    DTIC Science & Technology

    1978-12-29

    ANALYSIS 425 D-3 INFORMATION SCIENCi AND DECISION ANALYSIS 427 D-4 MAINTENANCE THEORY AND PHILOSOPHY 430 D-5 MAINTENANCE APPLUCATIONS 436 D-6 A GUIDE...on-condition task. Unfor- tunately it is usually beyond human capability to look at a used part and determine what its likelihood of failure will be...be found in serviceable units that are removed at some specified age limit, but -t is generally beyond human capability to estimate from this early

  1. GMI1, a structural-maintenance-of-chromosomes-hinge domain-containing protein, is involved in somatic homologous recombination in Arabidopsis.

    PubMed

    Böhmdorfer, Gudrun; Schleiffer, Alexander; Brunmeir, Reinhard; Ferscha, Stefan; Nizhynska, Viktoria; Kozák, Jaroslav; Angelis, Karel J; Kreil, David P; Schweizer, Dieter

    2011-08-01

    DNA double-strand breaks (DSBs) pose one of the most severe threats to genome integrity, potentially leading to cell death. After detection of a DSB, the DNA damage and repair response is initiated and the DSB is repaired by non-homologous end joining and/or homologous recombination. Many components of these processes are still unknown in Arabidopsis thaliana. In this work, we characterized γ-irradiation and mitomycin C induced 1 (GMI1), a member of the SMC-hinge domain-containing protein family. RT-PCR analysis and promoter-GUS fusion studies showed that γ-irradiation, the radio-mimetic drug bleocin, and the DNA cross-linking agent mitomycin C strongly enhance GMI1 expression particularly in meristematic tissues. The induction of GMI1 by γ-irradiation depends on the signalling kinase Ataxia telangiectasia-mutated (ATM) but not on ATM and Rad3-related (ATR). Epistasis analysis of single and double mutants demonstrated that ATM acts upstream of GMI1 while the atr gmi1-2 double mutant was more sensitive than the respective single mutants. Comet assay revealed a reduced rate of DNA double-strand break repair in gmi1 mutants during the early recovery phase after exposure to bleocin. Moreover, the rate of homologous recombination of a reporter construct was strongly reduced in gmi1 mutant plants upon exposure to bleocin or mitomycin C. GMI1 is the first member of its protein family known to be involved in DNA repair. © 2011 The Authors. The Plant Journal © 2011 Blackwell Publishing Ltd.

  2. cAMP-dependent protein kinase and c-Jun N-terminal kinase mediate stathmin phosphorylation for the maintenance of interphase microtubules during osmotic stress.

    PubMed

    Yip, Yan Y; Yeap, Yvonne Y C; Bogoyevitch, Marie A; Ng, Dominic C H

    2014-01-24

    Dynamic microtubule changes after a cell stress challenge are required for cell survival and adaptation. Stathmin (STMN), a cytoplasmic microtubule-destabilizing phosphoprotein, regulates interphase microtubules during cell stress, but the signaling mechanisms involved are poorly defined. In this study ectopic expression of single alanine-substituted phospho-resistant mutants demonstrated that STMN Ser-38 and Ser-63 phosphorylation were specifically required to maintain interphase microtubules during hyperosmotic stress. STMN was phosphorylated on Ser-38 and Ser-63 in response to hyperosmolarity, heat shock, and arsenite treatment but rapidly dephosphorylated after oxidative stress treatment. Two-dimensional PAGE and Phos-tag gel analysis of stress-stimulated STMN phospho-isoforms revealed rapid STMN Ser-38 phosphorylation followed by subsequent Ser-25 and Ser-63 phosphorylation. Previously, we delineated stress-stimulated JNK targeting of STMN. Here, we identified cAMP-dependent protein kinase (PKA) signaling as responsible for stress-induced STMN Ser-63 phosphorylation. Increased cAMP levels induced by cholera toxin triggered potent STMN Ser-63 phosphorylation. Osmotic stress stimulated an increase in PKA activity and elevated STMN Ser-63 and CREB (cAMP-response element-binding protein) Ser-133 phosphorylation that was substantially attenuated by pretreatment with H-89, a PKA inhibitor. Interestingly, PKA activity and subsequent phosphorylation of STMN were augmented in the absence of JNK activation, indicating JNK and PKA pathway cross-talk during stress regulation of STMN. Taken together our study indicates that JNK- and PKA-mediated STMN Ser-38 and Ser-63 phosphorylation are required to preserve interphase microtubules in response to hyperosmotic stress.

  3. Mitochondrial DNA maintenance: an appraisal.

    PubMed

    Akhmedov, Alexander T; Marín-García, José

    2015-11-01

    Mitochondria play a crucial role in a variety of cellular processes ranging from energy metabolism, generation of reactive oxygen species (ROS), and Ca(2+) handling to stress responses, cell survival, and death. Malfunction of the organelle may contribute to the pathogenesis of neuromuscular disorders, cancer, premature aging, and cardiovascular diseases, including myocardial ischemia, cardiomyopathy, and heart failure. Mitochondria are unique as they contain their own genome organized into DNA-protein complexes, so-called mitochondrial nucleoids, along with multiprotein machineries, which promote mitochondrial DNA (mtDNA) replication, transcription, and repair. Although the organelle possesses almost all known nuclear DNA repair pathways, including base excision repair, mismatch repair, and recombinational repair, the proximity of mtDNA to the main sites of ROS production and the lack of protective histones may result in increased susceptibility to oxidative stress and other types of mtDNA damage. Defects in the components of these highly organized machineries, which mediate mtDNA maintenance (replication and repair), may result in accumulation of point mutations and/or deletions in mtDNA and decreased mtDNA copy number impairing mitochondrial function. This review will focus on the mechanisms of mtDNA maintenance with emphasis on the proteins implicated in these processes and their functional role in various disease conditions and aging.

  4. FGD maintenance guidelines. Volume 2: FGD maintenance information. Final report

    SciTech Connect

    Davidson, L.N.; Miller, G.P.; Wedig, C.P.

    1986-07-01

    The ''FGD Maintenance Guidelines'' was written to fill the need for maintenance information that applies specifically to flue gas desulfurization (FGD) systems. Previously there was no single source of data describing FGD maintenance. Usually maintenance managers relied on suppliers' operating and maintenance manuals, past FGD experience, or procedures from other areas of a power plant. There are two volumes in the Guidelines intended to assist utility personnel in planning and performing maintenance for FGD systems. Different aspects of maintenance are emphasized in each volume. Volume 1, FGD Maintenance Programs, provides guidance for supervisory personnel involved in planning maintenance. It describes the utility industry's experience with FGD maintenance programs, a procedure for organizing and managing maintenance programs, and ways to design FGD systems for maintainability. The section about implementing a maintenance program contains a detailed example to illustrate the procedure, based on experience of an actual operating FGD system. Volume 2, FGD Maintenance Information, has practical information, useful in understanding FGD systems and their maintenance needs. It describes the major types of FGD systems operating in the US and typical maintenance associated with each. Also, in this volume, there is information about maintenance needs and procedures for the most common types of FGD equipment. 21 refs., 48 figs., 3 tabs.

  5. In vitro maintenance of a new ovarian cancer cell line in protein-free media: a potential model for autonomous growth and tumor progression.

    PubMed

    Golombick, T; Bezwoda, W R

    1991-12-01

    A new cell line UWOV2 (pf) capable of long-term growth in the absence of any added serum protein, exogenous growth factor, insulin or transferrin, is described. The original cell line (UWOV2 and UWOV2 (sf), adapted to grow in serum-free conditions) was derived from the ascitic tumor of a patient with ovarian carcinoma. Under continuous culture conditions further adaptations have occurred enabling UWOV2 (pf) to maintain anchorage-dependent growth without requiring exogenous anchorage or growth factors. These cells produce a structured extracellular matrix which acts as an adhesive substrate for the UWOV2 (pf) cells themselves as well as for a number of other long-term cell lines including NRK and 3T3 cells. Furthermore, while UWOV2 (pf) cells produce a transforming growth factor beta (TGF beta)-like growth factor, they appear to be only partially dependent on autocrine growth stimulation, and other mechanisms for autonomous growth stimulation appear to exist. This cell line may be a useful model for the study of progressive growth autonomy in human tumors.

  6. Parathyroid hormone/parathyroid hormone-related protein receptor signaling is required for maintenance of the growth plate in postnatal life.

    PubMed

    Hirai, Takao; Chagin, Andrei S; Kobayashi, Tatsuya; Mackem, Susan; Kronenberg, Henry M

    2011-01-04

    Parathyroid hormone (PTH)-related protein (PTHrP), regulated by Indian hedgehog and acting through the PTH/PTHrP receptor (PPR), is crucial for normal cartilage development. These observations suggest a possible role of PPR signaling in the postnatal growth plate; however, the role of PPR signaling in postnatal chondrocytes is unknown. In this study, we have generated tamoxifen-inducible and cartilage-specific PPR KO mice to evaluate the physiological role of PPR signaling in postnatal chondrocytes. We found that inactivation of the PPR in chondrocytes postnatally leads to accelerated differentiation of chondrocytes, followed by disappearance of the growth plate. We also observed an increase of TUNEL-positive cells and activities of caspase-3 and caspase-9 in the growth plate, along with a decrease in phosphorylation of Bad at Ser155 in postnatal PPR KO mice. Administration of a low-phosphate diet, which prevents apoptosis of chondrocytes, prevented the disappearance of the growth plate. Taken together, these observations suggest that the major consequences of PPR activation are similar in both the fetal and postnatal growth plates. Moreover, chondrocyte apoptosis through the activation of a mitochondrial pathway may be involved in the process of premature disappearance of the growth plate by postnatal inactivation of the PPR in chondrocytes.

  7. Parathyroid hormone/parathyroid hormone-related protein receptor signaling is required for maintenance of the growth plate in postnatal life

    PubMed Central

    Hirai, Takao; Chagin, Andrei S.; Kobayashi, Tatsuya; Mackem, Susan; Kronenberg, Henry M.

    2011-01-01

    Parathyroid hormone (PTH)-related protein (PTHrP), regulated by Indian hedgehog and acting through the PTH/PTHrP receptor (PPR), is crucial for normal cartilage development. These observations suggest a possible role of PPR signaling in the postnatal growth plate; however, the role of PPR signaling in postnatal chondrocytes is unknown. In this study, we have generated tamoxifen-inducible and cartilage-specific PPR KO mice to evaluate the physiological role of PPR signaling in postnatal chondrocytes. We found that inactivation of the PPR in chondrocytes postnatally leads to accelerated differentiation of chondrocytes, followed by disappearance of the growth plate. We also observed an increase of TUNEL-positive cells and activities of caspase-3 and caspase-9 in the growth plate, along with a decrease in phosphorylation of Bad at Ser155 in postnatal PPR KO mice. Administration of a low-phosphate diet, which prevents apoptosis of chondrocytes, prevented the disappearance of the growth plate. Taken together, these observations suggest that the major consequences of PPR activation are similar in both the fetal and postnatal growth plates. Moreover, chondrocyte apoptosis through the activation of a mitochondrial pathway may be involved in the process of premature disappearance of the growth plate by postnatal inactivation of the PPR in chondrocytes. PMID:21173257

  8. Severe protein aggregate myopathy in a knockout mouse model points to an essential role of cofilin2 in sarcomeric actin exchange and muscle maintenance.

    PubMed

    Gurniak, Christine B; Chevessier, Frédéric; Jokwitz, Melanie; Jönsson, Friederike; Perlas, Emerald; Richter, Hendrik; Matern, Gabi; Boyl, Pietro Pilo; Chaponnier, Christine; Fürst, Dieter; Schröder, Rolf; Witke, Walter

    2014-01-01

    Mutations in the human actin depolymerizing factor cofilin2 result in an autosomal dominant form of nemaline myopathy. Here, we report on the targeted ablation of murine cofilin2, which leads to a severe skeletal muscle specific phenotype within the first two weeks after birth. Apart from skeletal muscle, cofilin2 is also expressed in heart and CNS, however the pathology was restricted to skeletal muscle. The two close family members of cofilin2 - ADF and cofilin1 - were co-expressed in muscle, but unable to compensate for the loss of cofilin2. While primary myofibril assembly and muscle development were unaffected in cofilin2 mutant mice, progressive muscle degeneration was observed between postnatal days 3 and 7. Muscle pathology was characterized by sarcoplasmic protein aggregates, fiber size disproportion, mitochondrial abnormalities and internal nuclei. The observed muscle pathology differed from nemaline myopathy, but showed combined features of actin-associated myopathy and myofibrillar myopathy. In cofilin2 mutant mice, the postnatal expression pattern and turnover of sarcomeric α-actin isoforms were altered. Levels of smooth muscle α-actin were increased and remained high in developing muscles, suggesting that cofilin2 plays a crucial role during the exchange of α-actin isoforms during the early postnatal remodeling of the sarcomere.

  9. Differing antidepressant maintenance methodologies.

    PubMed

    Safer, Daniel J

    2017-10-01

    The principle evidence that antidepressant medication (ADM) is an effective maintenance treatment for adults with major depressive disorder (MDD) is from placebo substitution trials. These trials enter responders from ADM efficacy trials into randomized, double-blind placebo-controlled (RDBPC) effectiveness trials to measure the rate of MDD relapse over time. However, other randomized maintenance trial methodologies merit consideration and comparison. A systematic review of ADM randomized maintenance trials included research reports from multiple databases. Relapse rate was the main effectiveness outcome assessed. Five ADM randomized maintenance methodologies for MDD responders are described and compared for outcome. These effectiveness trials include: placebo-substitution, ADM/placebo extension, ADM extension, ADM vs. psychotherapy, and treatment as usual. The placebo-substitution trials for those abruptly switched to placebo resulted in unusually high (46%) rates of relapse over 6-12months, twice the continuing ADM rate. These trials were characterized by selective screening, high attrition, an anxious anticipation of a switch to placebo, and a risk of drug withdrawal symptoms. Selectively screened ADM efficacy responders who entered into 4-12month extension trials experienced relapse rates averaging ~10% with a low attrition rate. Non-industry sponsored randomized trials of adults with multiple prior MDD episodes who were treated with ADM maintenance for 1-2years experienced relapse rates averaging 40%. Placebo substitution trial methodology represents only one approach to assess ADM maintenance. Antidepressant maintenance research for adults with MDD should be evaluated for industry sponsorship, attrition, the impact of the switch to placebo, and major relapse differences in MDD subpopulations. Copyright © 2017. Published by Elsevier Inc.

  10. Generation, modulation and maintenance of the plasma membrane asymmetric phospholipid composition in yeast cells during growth: their relation to surface potential and membrane protein activity.

    PubMed

    Cerbón, J; Calderón, V

    1995-04-12

    During growth a cyclic exposure of anionic phospholipids to the external surface of the plasma membrane was found. The surface charge density (sigma) increased gradually reaching a maximum in the first 5 h of growth and returned gradually to their initial value at the end of the logarithmic phase of growth (10-12 h). Phosphatidylinositol, that determines to a large extent the magnitude of the sigma, increased 83% in the yeast cells during the first 4 h of growth and returned gradually to their initial level at 10-12 h. During the stationary phase (12-24 h), both sigma and the anionic/zwitterionic phospholipid ratio, remained without any significant variation. The high-affinity H-linked glutamate transport system that behaves as a sensor of the changes in the membrane surface potential (phi) increased its activity in the first 5 h and then decreased it, following with great accuracy the sigma variations and remained without changes during the stationary phase of growth. The phosphatidylserine (PS) relative concentration in the cells (9.0%) did not significantly change during the whole growth curve, but their asymmetric distribution varied, contributing to the changes in sigma. PS facing the outer membrane surface increased 2.45-times during the first 5 h of growth and then returned to their original value at the end of the log phase (12 h). Phosphatidylcholine (PC) remained constant during the whole growth curve (50%), while phosphatidylethanolamine (PE) decreased 3-fold in the first 4 h and then increased to its original value at 10 h. Interestingly, PE at the outer membrane surface remained constant (3% of the total phospholipids) during the whole growth curve. During growth yeast cells change their phospholipid composition originating altered patterns of the plasma membrane phospholipid composition and IN-OUT distribution. This dynamic asymmetry is involved in the regulation of the surface potential and membrane protein activity.

  11. Implications of levels of serum mineral metabolism markers, albumin and C-reactive protein for treatment costs of patients on maintenance dialysis.

    PubMed

    Salonen, Tapani; Piirto, Juho; Reina, Tuomo; Saha, Heikki; Pasternack, Amos

    2007-01-01

    Secondary hyperparathyroidism, malnutrition and inflammation have been reported to associate with adverse outcomes in dialysis patients. However, little is known about the implications of these conditions for treatment costs. The cost data of all adult patients who had entered dialysis therapy at Tampere University Hospital between 1991 and 1996 and had remained on dialysis for at least 1 year were collected. results of measurements of parathyroid hormone (PTH), calcium, phosphorus, albumin and C-reactive protein (CRP) were obtained from the database of the hospital. Patients (n = 109), aged 57.0 +/- 14.9 years, included 57% men and 37% diabetics; 62% started on hemodialysis and 38% on peritoneal dialysis. Average daily costs were USD 161 (range 95-360). After controlling for patients' age, body mass index, gender, dialysis modality and primary renal disease, there was a positive correlation between average CRP and average costs and a negative correlation between albumin and costs. Correlations between mineral metabolism markers and costs were not found, but there was a trend towards lower cost among patients who achieved the Kidney Disease Outcomes Quality Initiative targets of calcium, phosphorus and PTH (USD 145 +/- 31) compared with those with nonoptimal levels (USD 165 +/- 48; p = 0.095). Costs of patients with at least one in-target PTH measurement were lower than costs of patients with constantly low PTH (USD 148 +/- 31 vs. 170 +/- 48; p = 0.01). Serum levels of albumin and CRP correlated with dialysis patients' treatment costs. Achieving the Kidney Disease Outcomes Quality Initiative targets may be associated with lower costs. Copyright 2007 S. Karger AG, Basel.

  12. Building Maintenance, Management, and Budgeting.

    ERIC Educational Resources Information Center

    Pawsey, M. R.

    1982-01-01

    Australian methods and formulas for funding building maintenance and management are outlined and found to be haphazard. Discussed are: ultimate costs of deferred maintenance, major plant replacements, life cycle costing, types of maintenance programs (including full preventive maintenance), use of computer programs for planning, and organization…

  13. Building Maintenance, Management, and Budgeting.

    ERIC Educational Resources Information Center

    Pawsey, M. R.

    1982-01-01

    Australian methods and formulas for funding building maintenance and management are outlined and found to be haphazard. Discussed are: ultimate costs of deferred maintenance, major plant replacements, life cycle costing, types of maintenance programs (including full preventive maintenance), use of computer programs for planning, and organization…

  14. Reliability Centered Maintenance - Methodologies

    NASA Technical Reports Server (NTRS)

    Kammerer, Catherine C.

    2009-01-01

    Journal article about Reliability Centered Maintenance (RCM) methodologies used by United Space Alliance, LLC (USA) in support of the Space Shuttle Program at Kennedy Space Center. The USA Reliability Centered Maintenance program differs from traditional RCM programs because various methodologies are utilized to take advantage of their respective strengths for each application. Based on operational experience, USA has customized the traditional RCM methodology into a streamlined lean logic path and has implemented the use of statistical tools to drive the process. USA RCM has integrated many of the L6S tools into both RCM methodologies. The tools utilized in the Measure, Analyze, and Improve phases of a Lean Six Sigma project lend themselves to application in the RCM process. All USA RCM methodologies meet the requirements defined in SAE JA 1011, Evaluation Criteria for Reliability-Centered Maintenance (RCM) Processes. The proposed article explores these methodologies.

  15. Genetic response to bacteriophage infection in Lactococcus lactis reveals a four-strand approach involving induction of membrane stress proteins, D-alanylation of the cell wall, maintenance of proton motive force, and energy conservation.

    PubMed

    Fallico, Vincenzo; Ross, R Paul; Fitzgerald, Gerald F; McAuliffe, Olivia

    2011-11-01

    In this study, whole-genome microarrays were used to gain insights into the global molecular response of Lactococcus lactis subsp. lactis IL1403 at an early stage of infection with the lytic phage c2. The bacterium differentially regulated the expression of 61 genes belonging to 14 functional categories, including cell envelope processes (12 genes), regulatory functions (11 genes), and carbohydrate metabolism (7 genes). The nature of these genes suggests a complex response involving four main mechanisms: (i) induction of membrane stress proteins, (ii) d-alanylation of cell wall lipoteichoic acids (LTAs), (iii) maintenance of the proton motive force (PMF), and (iv) energy conservation. The phage presence is sensed as a membrane stress in L. lactis subsp. lactis IL1403, which activated a cell wall-targeted response probably orchestrated by the concerted action of membrane phage shock protein C-like homologues, the global regulator SpxB, and the two-component system CesSR. The bacterium upregulated genes (ddl and dltABCD) responsible for incorporation of d-alanine esters into LTAs, an event associated with increased resistance to phage attack in Gram-positive bacteria. The expression of genes (yshC, citE, citF) affecting both PMF components was also regulated to restore the physiological PMF, which was disrupted following phage infection. While mobilizing the response to the phage-mediated stress, the bacterium activated an energy-saving program by repressing growth-related functions and switching to anaerobic respiration, probably to sustain the PMF and the overall cell response to phage. To our knowledge, this represents the first detailed description in L. lactis of the molecular mechanisms involved in the host response to the membrane perturbations mediated by phage infection.

  16. Baffle maintenance apparatus

    SciTech Connect

    Genter, J. B.; Quinn, G. J.; Shields, E. P.; Thomas, B. S.; Winkler, C. J.

    1983-12-20

    The baffle maintenance apparatus comprises a peening apparatus mounted on a movable carriage for reducing the width of a gap between two adjacent baffle plates in a nuclear reactor. The peening apparatus comprises an hydraulic hammer mounted on a pivotable plate that is capable of being positioned in proper relationship to the baffle plate for reducing the gap therebetween. The baffle maintenance apparatus may also comprise a gauging mechanism mounted on the carriage for determining the width of the gap between the baffle plates.

  17. DLG5 in cell polarity maintenance and cancer development.

    PubMed

    Liu, Jie; Li, Juan; Ren, Yu; Liu, Peijun

    2014-01-01

    Failure in establishment and maintenance of epithelial cell polarity contributes to tumorigenesis. Loss of expression and function of cell polarity proteins is directly related to epithelial cell polarity maintenance. The polarity protein discs large homolog 5 (DLG5) belongs to a family of molecular scaffolding proteins called Membrane Associated Guanylate Kinases (MAGUKs). As the other family members, DLG5 contains the multi-PDZ, SH3 and GUK domains. DLG5 has evolved in the same manner as DLG1 and ZO1, two well-studied MAGUKs proteins. Just like DLG1 and ZO1, DLG5 plays a role in cell migration, cell adhesion, precursor cell division, cell proliferation, epithelial cell polarity maintenance, and transmission of extracellular signals to the membrane and cytoskeleton. Since the roles of DLG5 in inflammatory bowel disease (IBD) and Crohn's disease (CD) have been reviewed, here, our review focuses on the roles of DLG5 in epithelial cell polarity maintenance and cancer development.

  18. Carnitine in maintenance hemodialysis patients.

    PubMed

    Guarnieri, Gianfranco

    2015-03-01

    Carnitine is a conditionally essential metabolite that plays a critical role in cell physiology. Carnitine is necessary for fatty acid transport to sites of beta-oxidation in the mitochondria, where it also helps to prevent organic acid accumulation. Because of these key regulatory functions, carnitine represents a crucial determinant of mitochondrial energy metabolism, whose deficiency may lead to metabolic and clinical disturbances. Loss of carnitine through dialytic membranes occurs in maintenance hemodialysis, resulting in potential carnitine depletion and relative increments of esterified carnitine forms. Carnitine supplementation has been reported to counteract some of these alterations and has been associated with some clinical benefits, such as enhanced response to erythropoietin as well as improvement in exercise tolerance, intradialytic symptom, hyperparathyroidism, insulin resistance, inflammatory and oxidant status, protein balance, lipid profile, cardiac function, and quality of life. Carnitine supplementation has an attractive theoretical rationale; however, there are no definitive supportive studies and conclusive evidence that L-carnitine supplementation in maintenance hemodialysis patients could improve these conditions. A trial of carnitine administration could be attempted for 6 to 12 months only in selected patients on dialysis who do not adequately respond to standard therapies, in the presence of symptomatology, and in conjunction with patient dialysis age and documented L-carnitine deficiency.

  19. Diesel Vehicle Maintenance Competencies.

    ERIC Educational Resources Information Center

    Braswell, Robert; And Others

    Designed to provide a model set of competencies, this manual presents tasks which were identified by employers, employees, and teachers as important in a postsecondary diesel vehicle maintenance curriculum. The tasks are divided into seven major component areas of instruction: chassis and suspension, diesel engines, diesel fuel, electrical,…

  20. Horticulture: Grounds Maintenance Employee.

    ERIC Educational Resources Information Center

    Sims, James; And Others

    The unit of individualized learning activities is designed to provide training in grounds maintenance. The materials in the unit are divided into two sections. The developmental or preliminary phase (15 pages) is for use by the instructor and includes brief descriptions of the job and of the student population, along with listings of the specific…

  1. Maintenance Crisis vs Solutions.

    ERIC Educational Resources Information Center

    Haggard, Susie

    Industrial maintenance in Northeast Georgia is facing an acute crisis. Contributing factors are economic development that is depleting the work force, aging of the population, downsizing of the military, and lack of technical school graduates. Solutions to the crisis fall into three categories: short-term, mid-term, and long-term. For short-term…

  2. Floors: Selection and Maintenance.

    ERIC Educational Resources Information Center

    Berkeley, Bernard

    Flooring for institutional, commercial, and industrial use is described with regard to its selection, care, and maintenance. The following flooring and subflooring material categories are discussed--(1) resilient floor coverings, (2) carpeting, (3) masonry floors, (4) wood floors, and (5) "formed-in-place floors". The properties, problems,…

  3. Floors: Care and Maintenance.

    ERIC Educational Resources Information Center

    Post Office Dept., Washington, DC.

    Guidelines, methods and policies regarding the care and maintenance of post office building floors are overviewed in this handbook. Procedures outlined are concerned with maintaining a required level of appearance without wasting manpower. Flooring types and characteristics and the particular cleaning requirements of each type are given along with…

  4. Operations and maintenance philosophy

    SciTech Connect

    DUNCAN, G.P.

    1999-10-28

    This Operations and Maintenance (O&M) Philosophy document is intended to establish a future O&M vision, with an increased focus on minimizing worker exposure, ensuring uninterrupted retrieval operations, and minimizing operation life-cycle cost. It is intended that this document would incorporate O&M lessons learned into on-going and future project upgrades.

  5. Care and Maintenance.

    ERIC Educational Resources Information Center

    Hampton, Carol D.; Hampton, Carolyn H.

    1980-01-01

    Described is a method for bringing the sea into the classroom by setting up a saltwater aquarium. Included is selection of an aquarium, filtering systems, water (whether natural salt or synthetic sea salts), bottom materials, setting up an aquarium, system stabilization, stocking an aquarium, and maintenance of the aquarium. (DS)

  6. Maintenance Crisis vs Solutions.

    ERIC Educational Resources Information Center

    Haggard, Susie

    Industrial maintenance in Northeast Georgia is facing an acute crisis. Contributing factors are economic development that is depleting the work force, aging of the population, downsizing of the military, and lack of technical school graduates. Solutions to the crisis fall into three categories: short-term, mid-term, and long-term. For short-term…

  7. Care and Maintenance.

    ERIC Educational Resources Information Center

    Hampton, Carol D.; Hampton, Carolyn H.

    1980-01-01

    Described is a method for bringing the sea into the classroom by setting up a saltwater aquarium. Included is selection of an aquarium, filtering systems, water (whether natural salt or synthetic sea salts), bottom materials, setting up an aquarium, system stabilization, stocking an aquarium, and maintenance of the aquarium. (DS)

  8. Electrical Maintenance Technician.

    ERIC Educational Resources Information Center

    Ohio State Univ., Columbus. Center on Education and Training for Employment.

    This document contains 30 units to consider for use in a tech prep competency profile for the occupation of electrical maintenance technician. All the units listed will not necessarily apply to every situation or tech prep consortium, nor will all the competencies within each unit be appropriate. Several units appear within each specific…

  9. School Maintenance Needs

    ERIC Educational Resources Information Center

    Erickson, Paul W.

    2011-01-01

    The United States is full of schools built in the 1950s and 60s that supported the boomer school-age enrollment increase. These schools, once beacons of the neighborhood, are 50 to 60 years old and susceptible to becoming the community "eyesore." Budgeting for maintenance was fairly systematic for school districts for the first 10 to 20…

  10. Home Maintenance Manual.

    ERIC Educational Resources Information Center

    Richmond, Jim; And Others

    This manual, written especially for the Navajo and Hopi Indian Relocation Commission, is a simply worded, step-by-step guide to home maintenance for new homeowners. It can be used for self-study or it can serve as instructional material for a training class on home ownership. The manual is organized in nine sections that cover the following…

  11. Floors: Selection and Maintenance.

    ERIC Educational Resources Information Center

    Berkeley, Bernard

    Flooring for institutional, commercial, and industrial use is described with regard to its selection, care, and maintenance. The following flooring and subflooring material categories are discussed--(1) resilient floor coverings, (2) carpeting, (3) masonry floors, (4) wood floors, and (5) "formed-in-place floors". The properties, problems,…

  12. Electrical Maintenance Technician.

    ERIC Educational Resources Information Center

    Ohio State Univ., Columbus. Center on Education and Training for Employment.

    This document contains 30 units to consider for use in a tech prep competency profile for the occupation of electrical maintenance technician. All the units listed will not necessarily apply to every situation or tech prep consortium, nor will all the competencies within each unit be appropriate. Several units appear within each specific…

  13. Playground Inspection & Maintenance.

    ERIC Educational Resources Information Center

    Leeds, Gerard

    People today demand a safer work environment and a safer play environment for children. Accidents such as broken arms are no longer accepted as an inevitable part of growing up. This paper presents recommendations for the maintenance of safe playground areas and equipment, covering three main areas: (1) inspections, which should follow a specified…

  14. School Maintenance Needs

    ERIC Educational Resources Information Center

    Erickson, Paul W.

    2011-01-01

    The United States is full of schools built in the 1950s and 60s that supported the boomer school-age enrollment increase. These schools, once beacons of the neighborhood, are 50 to 60 years old and susceptible to becoming the community "eyesore." Budgeting for maintenance was fairly systematic for school districts for the first 10 to 20…

  15. Diesel Vehicle Maintenance Competencies.

    ERIC Educational Resources Information Center

    Braswell, Robert; And Others

    Designed to provide a model set of competencies, this manual presents tasks which were identified by employers, employees, and teachers as important in a postsecondary diesel vehicle maintenance curriculum. The tasks are divided into seven major component areas of instruction: chassis and suspension, diesel engines, diesel fuel, electrical,…

  16. Wakata during TVIS maintenance

    NASA Image and Video Library

    2009-04-21

    ISS019-E-008758 (21 April 2009) --- Japan Aerospace Exploration Agency (JAXA) astronaut Koichi Wakata, Expedition 19/20 flight engineer, performs in-flight maintenance on the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station.

  17. Wakata during TVIS maintenance

    NASA Image and Video Library

    2009-04-21

    ISS019-E-008754 (21 April 2009) --- Japan Aerospace Exploration Agency (JAXA) astronaut Koichi Wakata, Expedition 19/20 flight engineer, performs in-flight maintenance on the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station.

  18. TVIS Inflight Maintenance

    NASA Image and Video Library

    2011-07-01

    ISS028-E-013758 (1 July 2011) --- Russian cosmonauts Sergei Volkov, Expedition 28 flight engineer; and Andrey Borisenko (mostly out of frame at left), commander, perform in-flight maintenance on the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station.

  19. Reiter performs TVIS maintenance

    NASA Image and Video Library

    2006-12-14

    ISS014-E-09897 (14 Dec. 2006) -- European Space Agency (ESA) astronaut Thomas Reiter, STS-116 mission specialist, works with the Treadmill Vibration Isolation System (TVIS) in the Zvezda Service Module of the International Space Station during in-flight maintenance (IFM) while Space Shuttle Discovery was docked with the station.

  20. Preventive Maintenance Process

    NASA Technical Reports Server (NTRS)

    Ciaruffoli, Veronica; Bramley, Craig; Matteson, Mike

    2001-01-01

    The Preventive Maintenance (PM) program at Stennis Space Center (SSC) evolved from an ineffective and poorly organized state to a highly organized state in which it became capable of tracking equipment, planning jobs with man hour estimates, and supporting outsourcing. This viewgraph presentation traces the steps the program took to improve itself.

  1. Ki-67, mini-chromosome maintenance 2 protein (MCM2) and geminin have no independent prognostic relevance for cancer-specific survival in surgically treated squamous cell carcinoma of the penis.

    PubMed

    May, Matthias; Burger, Maximilian; Otto, Wolfgang; Hakenberg, Oliver W; Wieland, Wolf F; May, Dieter; Hofstädter, Ferdinand; Götz, Stefanie; Niessl, Nina; Fritsche, Hans-Martin; Birnkammer, Kristina; Gilfrich, Christian; Peter, Julia; Jain, Anjun; Koch, Stefan; Lebentrau, Steffen; Riedmiller, Hubertus; Rössler, Wolfgang; Denzinger, Stefan; Brookman-May, Sabine; Gunia, Sven

    2013-08-01

    What's known on the subject? and what does the study add?: Only little and partly contradictory data are currently published about the prognostic role of immunohistochemically detectable proliferation-associated biomarkers in surgically treated squamous cell carcinoma of the penis (SCCP), and no data are available at present about their usefulness for refining the delineation between different Broders' grading categories (e.g. still G2 or just G3 SCCP?). Moreover, the accuracy of various conventional histopathological parameters for predicting cancer-specific survival (CSS) in surgically treated SCCP has not been systematically evaluated yet. Based on the so far largest study cohort encompassing 158 consecutive patients with surgically treated PSCCs characterised by means of a central histopathological review, our data add the following to the currently available literature: (i) Ki-67, mini-chromosome maintenance 2 protein (MCM2), and geminin indicate a more aggressive behaviour in SCPP but do not represent independent prognostic parameters in the multivariable analysis in terms of CSS, (ii) these three biomarkers are not helpful for refining the delineation between different Broders' grading categories at the immunohistochemical level, and (iii) the conventional histopathological parameters staging, grading, nodal involvement, and lymphovascular invasion are independent prognostic parameters that together achieve a predictive accuracy of 82% for CSS. To assess the role of cell proliferation-associated biomarkers to predict cancer-specific survival (CSS) in patients with surgically treated squamous cell carcinoma of the penis (SCCP). A multicentre study enrolling 158 consecutive patients with surgically treated SCCP was performed. After conducting a central histopathological review, the staining profiles of Ki-67, mini-chromosome maintenance 2 protein (MCM2) and geminin were evaluated for their correlation with conventional histopathological criteria and their

  2. Knockdown of Minichromosome Maintenance Proteins Inhibits Foci Forming of Mediator of DNA-Damage Checkpoint 1 in Response to DNA Damage in Human Esophageal Squamous Cell Carcinoma TE-1 Cells.

    PubMed

    Yu, Jinzhong; Wang, Ruijie; Wu, Jinfeng; Dang, Zhongqin; Zhang, Qinsheng; Li, Bo

    2016-10-01

    Esophageal squamous cell carcinoma (ESCC) has a high morbidity in China and its treatment depends greatly on adjuvant chemotherapy. However, DNA damage repair in cancer cells severely affects the outcome of treatment. This study investigated the potential mechanism regarding mediator of DNA-damage checkpoint 1 (MDC1) and minichromosome maintenance proteins (MCMs) during DNA damage in ESCC. Recombinant vectors of MDC1 and MCMs with tags were constructed and transfected into human ESCC cell line TE-1. Immunoprecipitation and mass spectrometry were performed to screen the MCMs interacting with MDC1, and direct interaction was confirmed by glutathione S-transferase (GST) pull-down assay in vitro. MCM2 and MCM6 were knocked down by shRNAs, after which chromatin fraction and foci forming of MDC1 upon bleomycin-induced DNA damage were examined. The results showed that MCM2/3/5/6 were immunoprecipitated by antibodies against the tag of MDC1 in TE-1 nuclei, and the GST pull-down assay indicated the direct interaction. Knockdown of MCM2 or MCM6 reduced the chromatin fraction of MDC1 according to Western blot results. Moreover, knockdown of MCM2 or MCM6 could significantly inhibit foci forming of MDC1 in TE-1 nuclei in response to bleomycin-induced DNA damage (p < 0.001). This study indicates the direct interaction between MDC1 and MCMs in TE-1 nuclei. Downregulation of MCMs can inhibit chromatin fraction and foci forming of MDC1 in TE-1 cells upon DNA damage, which suggests MCMs and MDC1 as potential targets to improve the outcome of chemotherapy in ESCC.

  3. The mitochondrial nucleoid protein, Mgm101p, of Saccharomyces cerevisiae is involved in the maintenance of rho(+) and ori/rep-devoid petite genomes but is not required for hypersuppressive rho(-) mtDNA.

    PubMed

    Zuo, Xiao Ming; Clark-Walker, G Desmond; Chen, Xin Jie

    2002-04-01

    The Saccharomyces cerevisiae MGM101 gene encodes a DNA-binding protein targeted to mitochondrial nucleoids. MGM101 is essential for maintenance of a functional rho(+) genome because meiotic segregants, with a disrupted mgm101 allele, cannot undergo more than 10 divisions on glycerol medium. Quantitative analysis of mtDNA copy number in a rho(+) strain carrying a temperature-sensitive allele, mgm101-1, revealed that the amount of mtDNA is halved each cell division upon a shift to the restrictive temperature. These data suggest that mtDNA replication is rapidly blocked in cells lacking MGM101. However, a small proportion of meiotic segregants, disrupted in MGM101, have rho(-) genomes that are stably maintained. Interestingly, all surviving rho(-) mtDNAs contain an ori/rep sequence. Disruption of MGM101 in hypersuppressive (HS) strains does not have a significant effect on the propagation of HS rho(-) mtDNA. However, in petites lacking an ori/rep, disruption of MGM101 leads to either a complete loss or a dramatically decreased stability of mtDNA. This discriminatory effect of MGM101 suggests that replication of rho(+) and ori/rep-devoid rho(-) mtDNAs is carried out by the same process. By contrast, the persistence of ori/rep-containing mtDNA in HS petites lacking MGM101 identifies a distinct replication pathway. The alternative mtDNA replication mechanism provided by ori/rep is independent of mitochondrial RNA polymerase encoded by RPO41 as a HS rho(-) genome is stably maintained in a mgm101, rpo41 double mutant.

  4. CFB refractory maintenance

    SciTech Connect

    Johnson, R.C.

    1994-12-31

    The CFB maintenance manager has to make rapid-fire decisions regarding refractory maintenance during short or unscheduled turnarounds. This presentation offers a hands-on approach to expedient refractory failure analyses with specific repair recommendations. Photographs of most typical CFB refractory failures and their structural repairs are discussed. The most reliable repairs can be expected by using the latest state-of-the-art refractory materials and installation techniques. Refractory materials are consumable; therefore minor repairs should always be conducted at the first opportunity; this will preclude future major repairs. During a short or unscheduled outage, major repairs should be confined to the specific structural repair site; the removal of good, serviceable refractory is unnecessary under these conditions.

  5. Terminal automation system maintenance

    SciTech Connect

    Coffelt, D.; Hewitt, J.

    1997-01-01

    Nothing has improved petroleum product loading in recent years more than terminal automation systems. The presence of terminal automation systems (TAS) at loading racks has increased operational efficiency and safety and enhanced their accounting and management capabilities. However, like all finite systems, they occasionally malfunction or fail. Proper servicing and maintenance can minimize this. And in the unlikely event a TAS breakdown does occur, prompt and effective troubleshooting can reduce its impact on terminal productivity. To accommodate around-the-clock loading at racks, increasingly unattended by terminal personnel, TAS maintenance, servicing and troubleshooting has become increasingly demanding. It has also become increasingly important. After 15 years of trial and error at petroleum and petrochemical storage and transfer terminals, a number of successful troubleshooting programs have been developed. These include 24-hour {open_quotes}help hotlines,{close_quotes} internal (terminal company) and external (supplier) support staff, and {open_quotes}layered{close_quotes} support. These programs are described.

  6. Flight Crew Health Maintenance

    NASA Technical Reports Server (NTRS)

    Gullett, C. C.

    1970-01-01

    The health maintenance program for commercial flight crew personnel includes diet, weight control, and exercise to prevent heart disease development and disability grounding. The very high correlation between hypertension and overweight in cardiovascular diseases significantly influences the prognosis for a coronary prone individual and results in a high rejection rate of active military pilots applying for civilian jobs. In addition to physical fitness the major items stressed in pilot selection are: emotional maturity, glucose tolerance, and family health history.

  7. Flight Crew Health Maintenance

    NASA Technical Reports Server (NTRS)

    Gullett, C. C.

    1970-01-01

    The health maintenance program for commercial flight crew personnel includes diet, weight control, and exercise to prevent heart disease development and disability grounding. The very high correlation between hypertension and overweight in cardiovascular diseases significantly influences the prognosis for a coronary prone individual and results in a high rejection rate of active military pilots applying for civilian jobs. In addition to physical fitness the major items stressed in pilot selection are: emotional maturity, glucose tolerance, and family health history.

  8. PREVENTIVE MAINTENANCE. HONEYWELL PLANNING GUIDE.

    ERIC Educational Resources Information Center

    Honeywell, Inc., Minneapolis, Minn.

    THIS HONEYWELL PAMPHLET DISCUSSES SOME ASPECTS OF PREVENTIVE MAINTENANCE OF AUTOMATIC CONTROLS, HEATING, VENTILATING, AND AIR CONDITIONING, AND COMPARES IN-PLANT WITH CONTRACT SERVICE, CONCLUDING THAT CONTRACT SERVICE IS PREFERABLE AND DESCRIBING A NUMBER OF MAINTENANCE PLANS WHICH THEY FURNISH. PREVENTIVE MAINTENANCE PROVIDES--(1) MORE EFFICIENT…

  9. AUTOMOTIVE DIESEL MAINTENANCE. PROGRAM OUTLINE.

    ERIC Educational Resources Information Center

    Human Engineering Inst., Cleveland, OH.

    INFORMATIONAL TOPICS COVERED IN THE TEXT MATERIALS AND SELF-INSTRUCTIONAL BRANCH PROGRAMED TRAINING FILMS FOR A 2-YEAR, 55 MODULE PROGRAM IN AUTOMOTIVE DIESEL MAINTENANCE ARE GIVEN. THE 30 MODULES FOR "AUTOMOTIVE DIESEL MAINTENANCE 1" ARE AVAILABLE AS VT 005 655 - VT 005 684, AND THE 25 MODULES FOR "AUTOMOTIVE DIESEL MAINTENANCE 2" ARE AVAILABLE…

  10. Management Aspects of Software Maintenance.

    DTIC Science & Technology

    1984-09-01

    TRADITIONAl METHODS ..... ............. 50 C. PARAMETRIC MODELS ..... .............. 53 D. ESTIMATING MAINTENANCE COSTS ... ......... 57 1. Planning...maintenance. 10 10A 4 7he extensive research dcne cn software development and on the management of the development process is only ncw begin- ning to...and external factors. C. GENEEAL PROCEDURE The procedure used was to research literature concerning software maintenance. Particular emphasis was

  11. AUTOMOTIVE DIESEL MAINTENANCE. PROGRAM OUTLINE.

    ERIC Educational Resources Information Center

    Human Engineering Inst., Cleveland, OH.

    INFORMATIONAL TOPICS COVERED IN THE TEXT MATERIALS AND SELF-INSTRUCTIONAL BRANCH PROGRAMED TRAINING FILMS FOR A 2-YEAR, 55 MODULE PROGRAM IN AUTOMOTIVE DIESEL MAINTENANCE ARE GIVEN. THE 30 MODULES FOR "AUTOMOTIVE DIESEL MAINTENANCE 1" ARE AVAILABLE AS VT 005 655 - VT 005 684, AND THE 25 MODULES FOR "AUTOMOTIVE DIESEL MAINTENANCE 2" ARE AVAILABLE…

  12. Proteins.

    ERIC Educational Resources Information Center

    Doolittle, Russell F.

    1985-01-01

    Examines proteins which give rise to structure and, by virtue of selective binding to other molecules, make genes. Binding sites, amino acids, protein evolution, and molecular paleontology are discussed. Work with encoding segments of deoxyribonucleic acid (exons) and noncoding stretches (introns) provides new information for hypotheses. (DH)

  13. Proteins.

    ERIC Educational Resources Information Center

    Doolittle, Russell F.

    1985-01-01

    Examines proteins which give rise to structure and, by virtue of selective binding to other molecules, make genes. Binding sites, amino acids, protein evolution, and molecular paleontology are discussed. Work with encoding segments of deoxyribonucleic acid (exons) and noncoding stretches (introns) provides new information for hypotheses. (DH)

  14. Protein

    USDA-ARS?s Scientific Manuscript database

    Proteins are the major structural and functional components of all cells in the body. They are macromolecules that comprise 1 or more chains of amino acids that vary in their sequence and length and are folded into specific 3-dimensional structures. The sizes and conformations of proteins, therefor...

  15. General aviation avionics equipment maintenance

    NASA Technical Reports Server (NTRS)

    Parker, C. D.; Tommerdahl, J. B.

    1978-01-01

    Maintenance of general aviation avionics equipment was investigated with emphasis on single engine and light twin engine general aviation aircraft. Factors considered include the regulatory agencies, avionics manufacturers, avionics repair stations, the statistical character of the general aviation community, and owners and operators. The maintenance, environment, and performance, repair costs, and reliability of avionics were defined. It is concluded that a significant economic stratification is reflected in the maintenance problems encountered, that careful attention to installations and use practices can have a very positive impact on maintenance problems, and that new technologies and a general growth in general aviation will impact maintenance.

  16. Paranal maintenance and CMMS experience

    NASA Astrophysics Data System (ADS)

    Montano, Nelson

    2004-10-01

    During the last four years of operations, low technical downtime has been one of the relevant records of the Paranal Observatory. From the beginning of the Very Large Telescope (VLT) project, European Southern Observatory (ESO) has considered the implementation of a proper maintenance strategy a fundamental point in order to ensure low technical down time and preserve the Observatory's assets. The implementation of the maintenance strategy was based on the following aspects: - Strong maintenance sense during the design stage. Line Replacement Unit (LRU) concept, standardization and modularity of the Observatory equipment - Creation of a dedicated team for Maintenance - The implementation of a Computerized Maintenance Management System After four operational years, the result of these aspects has exceeded the expectations; the Observatory has been operating with high availability under a sustainable strategy. The strengths of the maintenance strategy have been based on modern maintenance concepts applied by regular production companies, where any minute of down time involves high cost. The operation of the actual Paranal Maintenance System is based mainly on proactive activities, such as regular inspections, preventive maintenance (PM) and predictive maintenance (PdM) plans. Nevertheless, it has been necessary to implement a strong plan for corrective maintenance (CM). The Spare Parts Strategy has also been an important point linked to the Maintenance Strategy and CMMS implementation. At present, almost 4,000 items related to the Observatory spare parts are loaded into the CMMS database. Currently, we are studying the implementation of a Reliability Centered Maintenance (RCM) project in one of our critical systems The following document presents the actual status of the Paranal Maintenance Strategy and which have been the motivations to implement the established strategy.

  17. Protein

    MedlinePlus

    ... Search for: Harvard T.H. Chan School of Public Health Email People Departments Calendar Careers Give my.harvard ... Nutrition Source Harvard T.H. Chan School of Public Health > The Nutrition Source > What Should I Eat? > Protein ...

  18. Qualitative Maintenance Experience Handbook

    DTIC Science & Technology

    1975-10-20

    rdme hinge nonression springn. 7. Rem~ove 3 nuts, 1w- ashers , and bolts holding hinge pin halvers together. 8. Remove piD pin senuring iury strut and...bolt, nut. i,, asher , and pin. ONSTALLATTON: I. Reverse )’F -emov,,I. P. Aotuator is pr c-ad;Iusted to lenth and probably does not need adjust 3. TI...time in that syst-m and also permits malfunctions to be induces in a system not directly associated with the maintenance. [ ITI I - ___ ~ JaI , FLIGHT

  19. 14 CFR 43.9 - Content, form, and disposition of maintenance, preventive maintenance, rebuilding, and alteration...

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... maintenance, preventive maintenance, rebuilding, and alteration records (except inspections performed in..., PREVENTIVE MAINTENANCE, REBUILDING, AND ALTERATION § 43.9 Content, form, and disposition of maintenance, preventive maintenance, rebuilding, and alteration records (except inspections performed in accordance...

  20. 14 CFR 43.9 - Content, form, and disposition of maintenance, preventive maintenance, rebuilding, and alteration...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... maintenance, preventive maintenance, rebuilding, and alteration records (except inspections performed in..., PREVENTIVE MAINTENANCE, REBUILDING, AND ALTERATION § 43.9 Content, form, and disposition of maintenance, preventive maintenance, rebuilding, and alteration records (except inspections performed in accordance...

  1. 14 CFR 43.9 - Content, form, and disposition of maintenance, preventive maintenance, rebuilding, and alteration...

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... maintenance, preventive maintenance, rebuilding, and alteration records (except inspections performed in..., PREVENTIVE MAINTENANCE, REBUILDING, AND ALTERATION § 43.9 Content, form, and disposition of maintenance, preventive maintenance, rebuilding, and alteration records (except inspections performed in accordance with...

  2. Urinary tubular protein-based biomarkers in the rodent model of cisplatin nephrotoxicity: a comparative analysis of serum creatinine, renal histology, and urinary KIM-1, NGAL, and NAG in the initiation, maintenance, and recovery phases of acute kidney injury.

    PubMed

    Sinha, Vikash; Vence, Luis M; Salahudeen, Abdulla K

    2013-03-01

    Several biomarkers are becoming available for the early detection of acute kidney injury (AKI), but few have been directly compared. To compare urinary kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), and N-acetyl glucosaminidase (NAG) against serum creatinine and renal histological score in the initiation, maintenance, and recovery phases of cisplatin (CP)-induced AKI. Sprague-Dawley rats (300-350 g) were injected once through their tail veins with CP (CP group) at 5.5 mg/kg or with same volume of normal saline vehicle (Control group). Rats were euthanized at 2, 4, 6, 12, and 24 hours, and on days 2, 3, 6, and 10 (n = 12 in the CP group and n = 6 in the Control group at each time point), and urine, blood, and kidney samples were analyzed. A significant increase in serum creatinine was noted by day 3 in the CP group versus Control group [1.46 (0.12) vs 0.28 (0.03) mg/dL; mean (SE); P < 0.05]. The renal histology scores for brush border loss and tubular necrosis were significantly higher at 12 and 24 hours, respectively, in the CP group. Urinary kidney injury molecule-1 levels were significantly higher at 24 hours in the CP group than in the Control group [48.26 (13.13) vs 8.21 (3.31) pg/mg creatinine; P < 0.05] and remained elevated through day 10. Both urine NAG and NGAL levels were significantly higher by day 2 in the CP than in the Control group [NAG, 8.19 (0.82) vs 3.48 (0.40) pg/mg creatinine, P G 0.05; NGAL, 2911.80 (368.10) vs 1412.60 (250.20) pg/mg creatinine, P < 0.05]. Urinary NAG remained elevated for 6 days and NGAL for 3 days. Our study suggests a temporal hierarchy in the ability of certain urinary protein-based biomarkers to detect AKI after a well-defined tubular injury. Comparative analyses of urinary biomarkers are warranted in clinical settings such as patients receiving CP to discern the time course and pattern of expression.

  3. Maintenance and supply options

    NASA Technical Reports Server (NTRS)

    1988-01-01

    The object of the Maintenance and Supply Option was to develop a high level operational philosophy related to maintenance and supply operations and incorporate these concepts into the Lunar Base Study. Specific products to be generated during this task were three trade studies and a conceptual design of the Logistic Supply Module. The crew size study was performed to evaluate crew sizes from the baseline size of four to a crew size of eight and determine the preferred crew size. The second trade study was to determine the impact of extending surface stay times and recommend a preferred duration of stay time as a function of crew, consumables, and equipment support capabilities. The third trade study was an evaluation of packaging and storage methods to determine the preferred logistics approach to support the lunar base. A modified scenario was developed and served as the basis of the individual trade studies. Assumptions and guidelines were also developed from experience with Apollo programs, Space Shuttle operations, and Space Station studies. With this information, the trade studies were performed and a conceptual design for the Logistic Supply Module was developed.

  4. Towards automated traceability maintenance

    PubMed Central

    Mäder, Patrick; Gotel, Orlena

    2012-01-01

    Traceability relations support stakeholders in understanding the dependencies between artifacts created during the development of a software system and thus enable many development-related tasks. To ensure that the anticipated benefits of these tasks can be realized, it is necessary to have an up-to-date set of traceability relations between the established artifacts. This goal requires the creation of traceability relations during the initial development process. Furthermore, the goal also requires the maintenance of traceability relations over time as the software system evolves in order to prevent their decay. In this paper, an approach is discussed that supports the (semi-) automated update of traceability relations between requirements, analysis and design models of software systems expressed in the UML. This is made possible by analyzing change events that have been captured while working within a third-party UML modeling tool. Within the captured flow of events, development activities comprised of several events are recognized. These are matched with predefined rules that direct the update of impacted traceability relations. The overall approach is supported by a prototype tool and empirical results on the effectiveness of tool-supported traceability maintenance are provided. PMID:23471308

  5. Towards automated traceability maintenance.

    PubMed

    Mäder, Patrick; Gotel, Orlena

    2012-10-01

    Traceability relations support stakeholders in understanding the dependencies between artifacts created during the development of a software system and thus enable many development-related tasks. To ensure that the anticipated benefits of these tasks can be realized, it is necessary to have an up-to-date set of traceability relations between the established artifacts. This goal requires the creation of traceability relations during the initial development process. Furthermore, the goal also requires the maintenance of traceability relations over time as the software system evolves in order to prevent their decay. In this paper, an approach is discussed that supports the (semi-) automated update of traceability relations between requirements, analysis and design models of software systems expressed in the UML. This is made possible by analyzing change events that have been captured while working within a third-party UML modeling tool. Within the captured flow of events, development activities comprised of several events are recognized. These are matched with predefined rules that direct the update of impacted traceability relations. The overall approach is supported by a prototype tool and empirical results on the effectiveness of tool-supported traceability maintenance are provided.

  6. Remote maintenance monitoring system

    NASA Technical Reports Server (NTRS)

    Simpkins, Lorenz G. (Inventor); Owens, Richard C. (Inventor); Rochette, Donn A. (Inventor)

    1992-01-01

    A remote maintenance monitoring system retrofits to a given hardware device with a sensor implant which gathers and captures failure data from the hardware device, without interfering with its operation. Failure data is continuously obtained from predetermined critical points within the hardware device, and is analyzed with a diagnostic expert system, which isolates failure origin to a particular component within the hardware device. For example, monitoring of a computer-based device may include monitoring of parity error data therefrom, as well as monitoring power supply fluctuations therein, so that parity error and power supply anomaly data may be used to trace the failure origin to a particular plane or power supply within the computer-based device. A plurality of sensor implants may be rerofit to corresponding plural devices comprising a distributed large-scale system. Transparent interface of the sensors to the devices precludes operative interference with the distributed network. Retrofit capability of the sensors permits monitoring of even older devices having no built-in testing technology. Continuous real time monitoring of a distributed network of such devices, coupled with diagnostic expert system analysis thereof, permits capture and analysis of even intermittent failures, thereby facilitating maintenance of the monitored large-scale system.

  7. Asset Management and Facility Equipment Maintenance Nexus: Maintenance Effectiveness

    DTIC Science & Technology

    2013-09-01

    1000 Under Project #371013, “Linking FEM and Asset Management” ERDC TR-13-16 ii Abstract The US Army Corps of Engineers (USACE) has constructed...how data col- lected in Facility Equipment Maintenance ( FEM ) can be used to evaluate the effectiveness of maintenance to improve the condition of...lock infra- structure components. This includes analyzing how data already being col- lected in FEM can be used to evaluate maintenance effectiveness and

  8. Proteolysis in plasmid DNA stable maintenance in bacterial cells.

    PubMed

    Karlowicz, Anna; Wegrzyn, Katarzyna; Dubiel, Andrzej; Ropelewska, Malgorzata; Konieczny, Igor

    2016-07-01

    Plasmids, as extrachromosomal genetic elements, need to work out strategies that promote independent replication and stable maintenance in host bacterial cells. Their maintenance depends on constant formation and dissociation of nucleoprotein complexes formed on plasmid DNA. Plasmid replication initiation proteins (Rep) form specific complexes on direct repeats (iterons) localized within the plasmid replication origin. Formation of these complexes along with a strict control of Rep protein cellular concentration, quaternary structure, and activity, is essential for plasmid maintenance. Another important mechanism for maintenance of low-copy-number plasmids are the toxin-antitoxin (TA) post-segregational killing (psk) systems, which prevent plasmid loss from the bacterial cell population. In this mini review we discuss the importance of nucleoprotein complex processing by energy-dependent host proteases in plasmid DNA replication and plasmid type II toxin-antitoxin psk systems, and draw attention to the elusive role of DNA in this process.

  9. Drivers of Bacterial Maintenance and Minimal Energy Requirements

    PubMed Central

    Kempes, Christopher P.; van Bodegom, Peter M.; Wolpert, David; Libby, Eric; Amend, Jan; Hoehler, Tori

    2017-01-01

    Microbes maintain themselves through a variety of processes. Several of these processes can be reduced or shut down entirely when resource availability declines. In pure culture conditions with ample substrate supply, a relationship between the maximum growth rate and the energy invested in maintenance has been reported widely. However, at the other end of the resources spectrum, bacteria are so extremely limited by energy that no growth occurs and metabolism is constrained to the most essential functions only. These minimum energy requirements have been called the basal power requirement. While seemingly different from each other, both aspects are likely components of a continuum of regulated maintenance processes. Here, we analyze cross-species tradeoffs in cellular physiology over the range of bacterial size and energy expenditure and determine the contributions to maintenance metabolism at each point along the size-energy spectrum. Furthermore, by exploring the simplest bacteria within this framework– which are most affected by maintenance constraints– we uncover which processes become most limiting. For the smallest species, maintenance metabolism converges on total metabolism, where we predict that maintenance is dominated by the repair of proteins. For larger species the relative costs of protein repair decrease and maintenance metabolism is predicted to be dominated by the repair of RNA components. These results provide new insights into which processes are likely to be regulated in environments that are extremely limited by energy. PMID:28197128

  10. Drivers of Bacterial Maintenance and Minimal Energy Requirements.

    PubMed

    Kempes, Christopher P; van Bodegom, Peter M; Wolpert, David; Libby, Eric; Amend, Jan; Hoehler, Tori

    2017-01-01

    Microbes maintain themselves through a variety of processes. Several of these processes can be reduced or shut down entirely when resource availability declines. In pure culture conditions with ample substrate supply, a relationship between the maximum growth rate and the energy invested in maintenance has been reported widely. However, at the other end of the resources spectrum, bacteria are so extremely limited by energy that no growth occurs and metabolism is constrained to the most essential functions only. These minimum energy requirements have been called the basal power requirement. While seemingly different from each other, both aspects are likely components of a continuum of regulated maintenance processes. Here, we analyze cross-species tradeoffs in cellular physiology over the range of bacterial size and energy expenditure and determine the contributions to maintenance metabolism at each point along the size-energy spectrum. Furthermore, by exploring the simplest bacteria within this framework- which are most affected by maintenance constraints- we uncover which processes become most limiting. For the smallest species, maintenance metabolism converges on total metabolism, where we predict that maintenance is dominated by the repair of proteins. For larger species the relative costs of protein repair decrease and maintenance metabolism is predicted to be dominated by the repair of RNA components. These results provide new insights into which processes are likely to be regulated in environments that are extremely limited by energy.

  11. TOR links starvation responses to telomere length maintenance.

    PubMed

    Kupiec, Martin; Weisman, Ronit

    2012-06-15

    Telomeres are nucleoprotein structures that protect the ends of eukaryotic chromosomes and play important roles in ensuring the genome's integrity. Telomere length is maintained by complex mechanisms that ensure length homeostasis. Recent work has linked telomere length maintenance to the Tor protein kinases, which are central regulators of cellular growth. Here we summarize these results, which suggest a link between nutrient availability, telomere length maintenance and chronological lifespan.

  12. Harnessing the power of maintenance

    SciTech Connect

    Toomey, G.

    2006-03-15

    More electric utilities are realizing that structured, comprehensive and proactive maintenance strategies deliver uptime to their bottom line. The article presents four case studies illustrating some of the available solutions to coal and gas-fired plant maintenance. These were: a maintenance strategy review of five coal fired and 22 gas-fired units to identify the importance of equipment along with maintenance tasks; a predictive maintenance program monitoring vibration to detect machine fault conditions including bearing condition; setting up a network to monitor data from several coal- and gas-fired plants at one central location; and creating a maintenance strategy to cushion against the possibility of unscheduled downtime and imposed penalties. 2 photos.

  13. Risk-based maintenance modeling. Prioritization of maintenance importances and quantification of maintenance effectiveness

    SciTech Connect

    Vesely, W.E.; Rezos, J.T.

    1995-09-01

    This report describes methods for prioritizing the risk importances of maintenances using a Probabilistic Risk Assessment (PRA). Approaches then are described for quantifying their reliability and risk effects. Two different PRA importance measures, minimal cutset importances and risk reduction importances, were used to prioritize maintenances; the findings show that both give similar results if appropriate criteria are used. The justifications for the particular importance measures also are developed. The methods developed to quantify the reliability and risk effects of maintenance actions are extensions of the usual reliability models now used in PRAs. These extended models consider degraded states of the component, and quantify the benefits of maintenance in correcting degradations and preventing failures. The negative effects of maintenance, including downtimes, also are included. These models are specific types of Markov models. The data for these models can be obtained from plant maintenance logs and from the Nuclear Plant Reliability Data System (NPRDS). To explore the potential usefulness of these models, the authors analyzed a range of postulated values of input data. These models were used to examine maintenance effects on a components reliability and performance for various maintenance programs and component data. Maintenance schedules were analyzed to optimize the component`s availability. In specific cases, the effects of maintenance were found to be large.

  14. Maintenance Strategy Maximising Availability Rate

    NASA Astrophysics Data System (ADS)

    Rosiński, Adam

    2012-12-01

    Electronic equipment operates under various conditions. Due to characteristic nature of its applications (e.g. in transport), it should be highly reliable. Many years’ worth of observations show, those systems not only require their constituting parts to function up to par, but also their maintenance has to be efficiently managed. This paper presents maintenance strategies and particularly focuses on maintenance strategy enabling maximising the availability rate.

  15. Boiler Combustion Control Maintenance Manual.

    DTIC Science & Technology

    1983-03-01

    controller and the final control element, as shown in Figure 6-1. The maintenance basically involves the inspection of the poppet valve in the relay...routine maintenance of this component basically involves placing a few drops of light machine oil on the ball bearing fulcrum. The poppet valve should... Valve /Air Damper Positioners ................................ 6-2 7.0 ELECTRIC PARALLEL POSITIONING CONTROL MAINTENANCE ................ 7-1 8.0 MOBILE

  16. Buildings Lean Maintenance Implementation Model

    NASA Astrophysics Data System (ADS)

    Abreu, Antonio; Calado, João; Requeijo, José

    2016-11-01

    Nowadays, companies in global markets have to achieve high levels of performance and competitiveness to stay "alive".Within this assumption, the building maintenance cannot be done in a casual and improvised way due to the costs related. Starting with some discussion about lean management and building maintenance, this paper introduces a model to support the Lean Building Maintenance (LBM) approach. Finally based on a real case study from a Portuguese company, the benefits, challenges and difficulties are presented and discussed.

  17. DCSP hardware maintenance system

    SciTech Connect

    Pazmino, M.

    1995-11-01

    This paper discusses the necessary changes to be implemented on the hardware side of the DCSP database. DCSP is currently tracking hardware maintenance costs in six separate databases. The goal is to develop a system that combines all data and works off a single database. Some of the tasks that will be discussed in this paper include adding the capability for report generation, creating a help package and preparing a users guide, testing the executable file, and populating the new database with data taken from the old database. A brief description of the basic process used in developing the system will also be discussed. Conclusions about the future of the database and the delivery of the final product are then addressed, based on research and the desired use of the system.

  18. Towards Agile Ontology Maintenance

    NASA Astrophysics Data System (ADS)

    Luczak-Rösch, Markus

    Ontologies are an appropriate means to represent knowledge on the Web. Research on ontology engineering reached practices for an integrative lifecycle support. However, a broader success of ontologies in Web-based information systems remains unreached while the more lightweight semantic approaches are rather successful. We assume, paired with the emerging trend of services and microservices on the Web, new dynamic scenarios gain momentum in which a shared knowledge base is made available to several dynamically changing services with disparate requirements. Our work envisions a step towards such a dynamic scenario in which an ontology adapts to the requirements of the accessing services and applications as well as the user's needs in an agile way and reduces the experts' involvement in ontology maintenance processes.

  19. Insect maintenance and transmission.

    PubMed

    Kingdom, Heather

    2013-01-01

    Phytoplasmas are plant pathogens of huge economic importance due to responsibility for crop yield losses worldwide. Institutions around the world are trying to understand and control this yield loss at a time when food security is high on government agendas. In order to fully understand the mechanisms of phytoplasma infection and spread, more insect vector and phytoplasma colonies will need to be established for research worldwide. Rearing and study of these colonies is essential in the research and development of phytoplasma control measures. This chapter highlights general materials and methods for raising insect vector colonies and maintenance of phytoplasmas. Specific methods of rearing the maize leafhopper and maize bushy stunt phytoplasma and the aster leafhopper and aster yellows phytoplasma strain witches' broom are also included.

  20. TCMS operations and maintenance philosophy

    NASA Technical Reports Server (NTRS)

    Buehler, David P.; Griffin, Rock E.

    1992-01-01

    The purpose is to describe the basic philosophies of operating and maintaining the Test, Control, and Monitor System (TCMS) equipment. TCMS is a complex and sophisticated checkout system. Operations and maintenance processes developed to support it will be based upon current experience, but will be focused on the specific needs of TCMS in support of Space Station Freedom Program (SSFP) and related activities. An overview of the operations and maintenance goals and philosophies are presented. The assumptions, roles and responsibilities, concepts and interfaces for operation, on-line maintenance, off-line support, and Operations and Maintenance (O&M) personnel training on all TCMS equipment located at KSC are described.

  1. Space shuttle maintenance program planning document

    NASA Technical Reports Server (NTRS)

    Brown, D. V.

    1972-01-01

    A means for developing a space shuttle maintenance program which will be acceptable to the development centers, the operators (KSC and AF), and the manufacturer is presented. The general organization and decision processes for determining the essential scheduled maintenance requirements for the space shuttle orbiter are outlined. The development of initial scheduled maintenance programs is discussed. The remaining maintenance, that is non-scheduled or non-routine maintenance, is directed by the findings of the scheduled maintenance program and the normal operation of the shuttle. The remaining maintenance consists of maintenance actions to correct discrepancies noted during scheduled maintenance tasks, nonscheduled maintenance, normal operation, or condition monitoring.

  2. Modeling sRNA-Regulated Plasmid Maintenance

    PubMed Central

    Klumpp, Stefan

    2017-01-01

    We study a theoretical model for the toxin-antitoxin (hok/sok) mechanism for plasmid maintenance in bacteria. Toxin-antitoxin systems enforce the maintenance of a plasmid through post-segregational killing of cells that have lost the plasmid. Key to their function is the tight regulation of expression of a protein toxin by an sRNA antitoxin. Here, we focus on the nonlinear nature of the regulatory circuit dynamics of the toxin-antitoxin mechanism. The mechanism relies on a transient increase in protein concentration rather than on the steady state of the genetic circuit. Through a systematic analysis of the parameter dependence of this transient increase, we confirm some known design features of this system and identify new ones: for an efficient toxin-antitoxin mechanism, the synthesis rate of the toxin’s mRNA template should be lower that of the sRNA antitoxin, the mRNA template should be more stable than the sRNA antitoxin, and the mRNA-sRNA complex should be more stable than the sRNA antitoxin. Moreover, a short half-life of the protein toxin is also beneficial to the function of the toxin-antitoxin system. In addition, we study a therapeutic scenario in which a competitor mRNA is introduced to sequester the sRNA antitoxin, causing the toxic protein to be expressed. PMID:28085919

  3. Illinois Occupational Skill Standards: Industrial Maintenance General Maintenance Cluster.

    ERIC Educational Resources Information Center

    Illinois Occupational Skill Standards and Credentialing Council, Carbondale.

    These skill standards for the industrial maintenance general maintenance cluster are intended to be a guide to workforce preparation program providers in defining content for their programs and to employers to establish the skills and standards necessary for job acquisition. An introduction provides the Illinois perspective; Illinois Occupational…

  4. 14 CFR 121.367 - Maintenance, preventive maintenance, and alterations programs.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.367 Maintenance, preventive maintenance, and alterations..., preventive maintenance, and alterations that ensures that— (a) Maintenance, preventive......

  5. 14 CFR 135.433 - Maintenance and preventive maintenance training program.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Maintenance and preventive maintenance... SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.433 Maintenance and preventive maintenance training program. Each certificate holder or a person performing maintenance or...

  6. 14 CFR 121.367 - Maintenance, preventive maintenance, and alterations programs.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.367 Maintenance, preventive maintenance, and alterations..., preventive maintenance, and alterations that ensures that— (a) Maintenance, preventive......

  7. 14 CFR 121.367 - Maintenance, preventive maintenance, and alterations programs.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.367 Maintenance, preventive maintenance, and alterations..., preventive maintenance, and alterations that ensures that— (a) Maintenance, preventive......

  8. 14 CFR 121.367 - Maintenance, preventive maintenance, and alterations programs.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.367 Maintenance, preventive maintenance, and alterations..., preventive maintenance, and alterations that ensures that— (a) Maintenance, preventive......

  9. 14 CFR 121.367 - Maintenance, preventive maintenance, and alterations programs.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.367 Maintenance, preventive maintenance, and alterations..., preventive maintenance, and alterations that ensures that— (a) Maintenance, preventive......

  10. Landscaping With Maintenance in Mind.

    ERIC Educational Resources Information Center

    Sorensen, Randy

    2000-01-01

    Examines school ground landscape design that enhances attractive of the school and provides for easier maintenance. Landscape design issues discussed include choice of grass, trees, and shrubs; irrigation; and safety and access. Other considerations for lessening maintenance problems for facility managers are also highlighted. (GR)

  11. Preservation and Maintenance of Maps

    ERIC Educational Resources Information Center

    Capps, Marie T.

    1972-01-01

    The problems of storage and maintenance which confront the map librarian are discussed. Included are the causes of map damage and deterioration, methods of detection and correction, and suggestions of further measures for optimum preservation. Useful guides on preservation and maintenance are cited. (7 references) (Author/NH)

  12. Automated System Programs Preventive Maintenance.

    ERIC Educational Resources Information Center

    Locke, Richard C.

    1987-01-01

    A preventive maintenance system provides for the monitoring and inspection of school building elements in a programmed way through an automatic checklist. Utility cost savings are expected along with reduction of travel and wait time, and measurable standards of performance for all maintenance and repair work. (MLF)

  13. Preventive Maintenance Handbook. Audiovisual Equipment.

    ERIC Educational Resources Information Center

    Educational Products Information Exchange Inst., Stony Brook, NY.

    The preventive maintenance system for audiovisual equipment presented in this handbook is designed by specialists so that it can be used by nonspecialists in school sites. The report offers specific advice on saftey factors and also lists major problems that should not be handled by nonspecialists. Other aspects of a preventive maintenance system…

  14. Modern Method for Preventive Maintenance

    ERIC Educational Resources Information Center

    Wilson, Howard D.

    1974-01-01

    Describes a system at Michigan State University that comprises 16 separate programs and schedules 25,000 manhours of preventive maintenance. With information about preventive maintenance for over 100,000 units stored in its computer, the university saves personnel, time, and energy. (Author/MLF)

  15. Automotive Engine Maintenance and Repair.

    ERIC Educational Resources Information Center

    Marine Corps Inst., Washington, DC.

    This correspondence course, originally developed for the Marine Corps, is designed to provide students with an understanding of automotive engine maintenance and repair. The course contains six study units covering automotive engine maintenance and repair; design classification; engine malfunction, diagnosis, and repair; engine disassembly; engine…

  16. Truth Maintenance in Automatic Planning.

    DTIC Science & Technology

    The objective of this project was to explore the usefulness of incorporating truth maintenance system (TMS) technology into the design of planning...in the report. The six appendices describe the underlying research contributing to the design of the prototype system. Keywords: Truth maintenance, Planning search, Replanning, Nonmonotonic reasoning, Defeasible reasoning.

  17. Achieving world class maintenance status

    SciTech Connect

    Tomlingson, P.D.

    2007-08-15

    The article written by a management consultant, discusses the art of successful planning and operation of maintenance in mines considering factors such as benchmaking, key performance indices (KPIs) and frequency of procedures which can help achieve 'world class maintenance'. 1 fig.

  18. Landscaping With Maintenance in Mind.

    ERIC Educational Resources Information Center

    Sorensen, Randy

    2000-01-01

    Examines school ground landscape design that enhances attractive of the school and provides for easier maintenance. Landscape design issues discussed include choice of grass, trees, and shrubs; irrigation; and safety and access. Other considerations for lessening maintenance problems for facility managers are also highlighted. (GR)

  19. Preventative maintenance for control systems

    SciTech Connect

    Corbett, G.O.

    1985-05-01

    Preventative maintenance of the control systems should be performed on a required interval. Most manufacturers specify six to twelve month intervals. However, inspections can often be accomplished when the control units or mechanical systems are out of service for a malfunction repair. A procedural checklist for proper maintenance is given.

  20. Cell Maintenance Systems

    NASA Technical Reports Server (NTRS)

    Morrison, D. R.

    1985-01-01

    Living human cells require attachment to a suitable surface and special culture conditions in order to grow. These requirements are modified and amplified when cells are taken into a weightless environment. Special handling and maintenance systems are required for routine laboratory procedures conducted in the Orbiter and in the Spacelab. Methods were developed to maintain cells in special incubators designed for the Orbiter middeck, however, electrophoresis and other experiments require cells to be harvested off of the culture substrate before they can be processed or used. The cell transport assembly (CTA) was flown on STS-8, and results show that improvements are required to maintain adequate numbers of cells in this device longer than 48 hours. The life sciences middeck centrifuge probably can be used, but modifications will be required to transfer cells from the CTA and keep the cells sterile. Automated systems such as the Skylab SO-15 flight hardware and crew operated systems are being evaluated for use on the Space Shuttle, Spacelab, and Space Station research modules.

  1. Robots for Aircraft Maintenance

    NASA Technical Reports Server (NTRS)

    1993-01-01

    Marshall Space Flight Center charged USBI (now Pratt & Whitney) with the task of developing an advanced stripping system based on hydroblasting to strip paint and thermal protection material from Space Shuttle solid rocket boosters. A robot, mounted on a transportable platform, controls the waterjet angle, water pressure and flow rate. This technology, now known as ARMS, has found commercial applications in the removal of coatings from jet engine components. The system is significantly faster than manual procedures and uses only minimal labor. Because the amount of "substrate" lost is minimal, the life of the component is extended. The need for toxic chemicals is reduced, as is waste disposal and human protection equipment. Users of the ARMS work cell include Delta Air Lines and the Air Force, which later contracted with USBI for development of a Large Aircraft Paint Stripping system (LARPS). LARPS' advantages are similar to ARMS, and it has enormous potential in military and civil aircraft maintenance. The technology may also be adapted to aircraft painting, aircraft inspection techniques and paint stripping of large objects like ships and railcars.

  2. Symbolic Constraint Maintenance Grid

    NASA Technical Reports Server (NTRS)

    James, Mark

    2006-01-01

    Version 3.1 of Symbolic Constraint Maintenance Grid (SCMG) is a software system that provides a general conceptual framework for utilizing pre-existing programming techniques to perform symbolic transformations of data. SCMG also provides a language (and an associated communication method and protocol) for representing constraints on the original non-symbolic data. SCMG provides a facility for exchanging information between numeric and symbolic components without knowing the details of the components themselves. In essence, it integrates symbolic software tools (for diagnosis, prognosis, and planning) with non-artificial-intelligence software. SCMG executes a process of symbolic summarization and monitoring of continuous time series data that are being abstractly represented as symbolic templates of information exchange. This summarization process enables such symbolic- reasoning computing systems as artificial- intelligence planning systems to evaluate the significance and effects of channels of data more efficiently than would otherwise be possible. As a result of the increased efficiency in representation, reasoning software can monitor more channels and is thus able to perform monitoring and control functions more effectively.

  3. 14 CFR 91.417 - Maintenance records.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance... specified in paragraph (b) of this section: (1) Records of the maintenance, preventive maintenance, and... 14 Aeronautics and Space 2 2012-01-01 2012-01-01 false Maintenance records. 91.417 Section...

  4. 14 CFR 91.417 - Maintenance records.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance... specified in paragraph (b) of this section: (1) Records of the maintenance, preventive maintenance, and... 14 Aeronautics and Space 2 2011-01-01 2011-01-01 false Maintenance records. 91.417 Section...

  5. 14 CFR 91.417 - Maintenance records.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance... specified in paragraph (b) of this section: (1) Records of the maintenance, preventive maintenance, and... 14 Aeronautics and Space 2 2014-01-01 2014-01-01 false Maintenance records. 91.417 Section...

  6. 14 CFR 91.417 - Maintenance records.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance... specified in paragraph (b) of this section: (1) Records of the maintenance, preventive maintenance, and... 14 Aeronautics and Space 2 2013-01-01 2013-01-01 false Maintenance records. 91.417 Section...

  7. Operational Interventions to Maintenance Error

    NASA Technical Reports Server (NTRS)

    Kanki, Barbara G.; Walter, Diane; Dulchinos, VIcki

    1997-01-01

    A significant proportion of aviation accidents and incidents are known to be tied to human error. However, research of flight operational errors has shown that so-called pilot error often involves a variety of human factors issues and not a simple lack of individual technical skills. In aircraft maintenance operations, there is similar concern that maintenance errors which may lead to incidents and accidents are related to a large variety of human factors issues. Although maintenance error data and research are limited, industry initiatives involving human factors training in maintenance have become increasingly accepted as one type of maintenance error intervention. Conscientious efforts have been made in re-inventing the team7 concept for maintenance operations and in tailoring programs to fit the needs of technical opeRAtions. Nevertheless, there remains a dual challenge: 1) to develop human factors interventions which are directly supported by reliable human error data, and 2) to integrate human factors concepts into the procedures and practices of everyday technical tasks. In this paper, we describe several varieties of human factors interventions and focus on two specific alternatives which target problems related to procedures and practices; namely, 1) structured on-the-job training and 2) procedure re-design. We hope to demonstrate that the key to leveraging the impact of these solutions comes from focused interventions; that is, interventions which are derived from a clear understanding of specific maintenance errors, their operational context and human factors components.

  8. Operational Interventions to Maintenance Error

    NASA Technical Reports Server (NTRS)

    Kanki, Barbara G.; Walter, Diane; Dulchinos, VIcki

    1997-01-01

    A significant proportion of aviation accidents and incidents are known to be tied to human error. However, research of flight operational errors has shown that so-called pilot error often involves a variety of human factors issues and not a simple lack of individual technical skills. In aircraft maintenance operations, there is similar concern that maintenance errors which may lead to incidents and accidents are related to a large variety of human factors issues. Although maintenance error data and research are limited, industry initiatives involving human factors training in maintenance have become increasingly accepted as one type of maintenance error intervention. Conscientious efforts have been made in re-inventing the team7 concept for maintenance operations and in tailoring programs to fit the needs of technical opeRAtions. Nevertheless, there remains a dual challenge: 1) to develop human factors interventions which are directly supported by reliable human error data, and 2) to integrate human factors concepts into the procedures and practices of everyday technical tasks. In this paper, we describe several varieties of human factors interventions and focus on two specific alternatives which target problems related to procedures and practices; namely, 1) structured on-the-job training and 2) procedure re-design. We hope to demonstrate that the key to leveraging the impact of these solutions comes from focused interventions; that is, interventions which are derived from a clear understanding of specific maintenance errors, their operational context and human factors components.

  9. 340 Facility maintenance implementation plan

    SciTech Connect

    1995-03-01

    This Maintenance Implementation Plan (MIP) has been developed for maintenance functions associated with the 340 Facility. This plan is developed from the guidelines presented by Department of Energy (DOE) Order 4330.4B, Maintenance Management Program (DOE 1994), Chapter II. The objective of this plan is to provide baseline information for establishing and identifying Westinghouse Hanford Company (WHC) conformance programs and policies applicable to implementation of DOE order 4330.4B guidelines. In addition, this maintenance plan identifies the actions necessary to develop a cost-effective and efficient maintenance program at the 340 Facility. Primary responsibility for the performance and oversight of maintenance activities at the 340 Facility resides with Westinghouse Hanford Company (WHC). Maintenance at the 340 Facility is performed by ICF-Kaiser Hanford (ICF-KH) South Programmatic Services crafts persons. This 340 Facility MIP provides interface requirements and responsibilities as they apply specifically to the 340 Facility. This document provides an implementation schedule which has been developed for items considered to be deficient or in need of improvement. The discussion sections, as applied to implementation at the 340 Facility, have been developed from a review of programs and practices utilizing the graded approach. Biennial review and additional reviews are conducted as significant programmatic and mission changes are made. This document is revised as necessary to maintain compliance with DOE requirements.

  10. 14 CFR 43.9 - Content, form, and disposition of maintenance, preventive maintenance, rebuilding, and alteration...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT MAINTENANCE... preventive maintenance, rebuilds, or alters an aircraft, airframe, aircraft engine, propeller, appliance, or... maintenance, preventive maintenance, rebuilding, and alteration records (except inspections performed in...

  11. 14 CFR 43.9 - Content, form, and disposition of maintenance, preventive maintenance, rebuilding, and alteration...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT MAINTENANCE... preventive maintenance, rebuilds, or alters an aircraft, airframe, aircraft engine, propeller, appliance, or... maintenance, preventive maintenance, rebuilding, and alteration records (except inspections performed in...

  12. 14 CFR 43.3 - Persons authorized to perform maintenance, preventive maintenance, rebuilding, and alterations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT MAINTENANCE, PREVENTIVE MAINTENANCE, REBUILDING, AND... maintain, rebuild, alter, or perform preventive maintenance on an aircraft, airframe, aircraft engine... under part 61 may perform preventive maintenance on any aircraft owned or operated by that pilot which...

  13. Deferred School Maintenance Creates National Crises.

    ERIC Educational Resources Information Center

    Geiger, Philip E.

    2002-01-01

    Describes the cost and causes of the school maintenance "crises"; lists seven questions to determine if a school district has a quality maintenance program; describes consequences of deferred school maintenance in Yuma (Arizona) Union High School District. (PKP)

  14. 14 CFR 135.425 - Maintenance, preventive maintenance, and alteration programs.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.425 Maintenance, preventive maintenance, and alteration programs. Each certificate holder shall have an inspection program and... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Maintenance, preventive maintenance, and...

  15. 14 CFR 135.423 - Maintenance, preventive maintenance, and alteration organization.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.423 Maintenance, preventive maintenance, and alteration organization. (a) Each certificate holder that performs any of its... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Maintenance, preventive maintenance, and...

  16. 14 CFR 135.437 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.437 Authority to perform and approve maintenance, preventive maintenance, and alterations... maintenance, preventive maintenance, and alterations. 135.437 Section 135.437 Aeronautics and Space FEDERAL...

  17. 14 CFR 135.423 - Maintenance, preventive maintenance, and alteration organization.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.423 Maintenance, preventive maintenance, and alteration organization. (a) Each certificate holder that performs any of its... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Maintenance, preventive maintenance, and...

  18. 14 CFR 135.425 - Maintenance, preventive maintenance, and alteration programs.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.425 Maintenance, preventive maintenance, and alteration programs. Each certificate holder shall have an inspection program and... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Maintenance, preventive maintenance, and...

  19. 14 CFR 135.437 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.437 Authority to perform and approve maintenance, preventive maintenance, and alterations... maintenance, preventive maintenance, and alterations. 135.437 Section 135.437 Aeronautics and Space FEDERAL...

  20. 14 CFR 121.379 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... maintenance, preventive maintenance, and alterations. 121.379 Section 121.379 Aeronautics and Space FEDERAL... OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.379 Authority to perform and approve maintenance, preventive maintenance, and alterations. (a) A certificate holder may perform, or it may...

  1. 14 CFR 121.379 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... maintenance, preventive maintenance, and alterations. 121.379 Section 121.379 Aeronautics and Space FEDERAL... OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.379 Authority to perform and approve maintenance, preventive maintenance, and alterations. (a) A certificate holder may perform, or it may...

  2. 14 CFR 121.365 - Maintenance, preventive maintenance, and alteration organization.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.365 Maintenance, preventive maintenance, and alteration...), preventive maintenance, or alterations, and each person with whom it arranges for the performance of...

  3. 14 CFR 135.437 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... maintenance, preventive maintenance, and alterations. 135.437 Section 135.437 Aeronautics and Space FEDERAL... AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.437 Authority to perform and approve maintenance, preventive maintenance, and...

  4. 14 CFR 135.423 - Maintenance, preventive maintenance, and alteration organization.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Maintenance, preventive maintenance, and... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.423 Maintenance, preventive maintenance, and alteration organization. (a) Each certificate holder that performs any of...

  5. 14 CFR 135.425 - Maintenance, preventive maintenance, and alteration programs.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Maintenance, preventive maintenance, and... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.425 Maintenance, preventive maintenance, and alteration programs. Each certificate holder shall have an inspection program...

  6. 14 CFR 121.377 - Maintenance and preventive maintenance personnel duty time limitations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Maintenance and preventive maintenance..., Preventive Maintenance, and Alterations § 121.377 Maintenance and preventive maintenance personnel duty time limitations. Within the United States, each certificate holder (or person performing maintenance or...

  7. 14 CFR 121.377 - Maintenance and preventive maintenance personnel duty time limitations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Maintenance and preventive maintenance..., Preventive Maintenance, and Alterations § 121.377 Maintenance and preventive maintenance personnel duty time limitations. Within the United States, each certificate holder (or person performing maintenance or...

  8. 14 CFR 135.437 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... maintenance, preventive maintenance, and alterations. 135.437 Section 135.437 Aeronautics and Space FEDERAL... AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.437 Authority to perform and approve maintenance, preventive maintenance, and...

  9. 14 CFR 121.375 - Maintenance and preventive maintenance training program.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Maintenance and preventive maintenance... OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.375 Maintenance and preventive maintenance training program....

  10. 14 CFR 135.425 - Maintenance, preventive maintenance, and alteration programs.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Maintenance, preventive maintenance, and... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.425 Maintenance, preventive maintenance, and alteration programs. Each certificate holder shall have an inspection program...

  11. 14 CFR 121.379 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... maintenance, preventive maintenance, and alterations. 121.379 Section 121.379 Aeronautics and Space FEDERAL... OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.379 Authority to perform and approve maintenance, preventive maintenance, and alterations. (a) A certificate holder may perform, or it may...

  12. 14 CFR 135.437 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... maintenance, preventive maintenance, and alterations. 135.437 Section 135.437 Aeronautics and Space FEDERAL... AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.437 Authority to perform and approve maintenance, preventive maintenance, and...

  13. 14 CFR 121.375 - Maintenance and preventive maintenance training program.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Maintenance and preventive maintenance... OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.375 Maintenance and preventive maintenance training program....

  14. 14 CFR 121.379 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... maintenance, preventive maintenance, and alterations. 121.379 Section 121.379 Aeronautics and Space FEDERAL... OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.379 Authority to perform and approve maintenance, preventive maintenance, and alterations. (a) A certificate holder may perform, or it may...

  15. 14 CFR 121.377 - Maintenance and preventive maintenance personnel duty time limitations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Maintenance and preventive maintenance..., Preventive Maintenance, and Alterations § 121.377 Maintenance and preventive maintenance personnel duty time limitations. Within the United States, each certificate holder (or person performing maintenance or...

  16. 14 CFR 121.365 - Maintenance, preventive maintenance, and alteration organization.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.365 Maintenance, preventive maintenance, and alteration...), preventive maintenance, or alterations, and each person with whom it arranges for the performance of...

  17. 14 CFR 121.365 - Maintenance, preventive maintenance, and alteration organization.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.365 Maintenance, preventive maintenance, and alteration...), preventive maintenance, or alterations, and each person with whom it arranges for the performance of...

  18. 14 CFR 121.375 - Maintenance and preventive maintenance training program.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Maintenance and preventive maintenance... OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.375 Maintenance and preventive maintenance training program....

  19. 14 CFR 135.423 - Maintenance, preventive maintenance, and alteration organization.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Maintenance, preventive maintenance, and... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.423 Maintenance, preventive maintenance, and alteration organization. (a) Each certificate holder that performs any of...

  20. 14 CFR 121.379 - Authority to perform and approve maintenance, preventive maintenance, and alterations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... maintenance, preventive maintenance, and alterations. 121.379 Section 121.379 Aeronautics and Space FEDERAL... OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.379 Authority to perform and approve maintenance, preventive maintenance, and alterations. (a) A certificate holder may perform, or it may...

  1. 14 CFR 135.425 - Maintenance, preventive maintenance, and alteration programs.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Maintenance, preventive maintenance, and... ON BOARD SUCH AIRCRAFT Maintenance, Preventive Maintenance, and Alterations § 135.425 Maintenance, preventive maintenance, and alteration programs. Each certificate holder shall have an inspection program...

  2. 14 CFR 121.375 - Maintenance and preventive maintenance training program.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Maintenance and preventive maintenance... OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.375 Maintenance and preventive maintenance training program....

  3. 14 CFR 121.365 - Maintenance, preventive maintenance, and alteration organization.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Maintenance, preventive maintenance, and..., Preventive Maintenance, and Alterations § 121.365 Maintenance, preventive maintenance, and alteration...), preventive maintenance, or alterations, and each person with whom it arranges for the performance of...

  4. 14 CFR 121.375 - Maintenance and preventive maintenance training program.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Maintenance and preventive maintenance... OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Maintenance, Preventive Maintenance, and Alterations § 121.375 Maintenance and preventive maintenance training program....

  5. 14 CFR 121.377 - Maintenance and preventive maintenance personnel duty time limitations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Maintenance and preventive maintenance..., Preventive Maintenance, and Alterations § 121.377 Maintenance and preventive maintenance personnel duty time limitations. Within the United States, each certificate holder (or person performing maintenance or...

  6. 14 CFR 121.377 - Maintenance and preventive maintenance personnel duty time limitations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Maintenance and preventive maintenance..., Preventive Maintenance, and Alterations § 121.377 Maintenance and preventive maintenance personnel duty time limitations. Within the United States, each certificate holder (or person performing maintenance or...

  7. Managing Custodial and Maintenance Staffs.

    ERIC Educational Resources Information Center

    Fickes, Michael

    2001-01-01

    Presents some basic maintenance management techniques that can help schools meet their budgets, preserve staffing levels, meet productivity needs, and sustain quality services. Tips for staff recruitment, training, and retention are explored. (GR)

  8. Endeavour Return to Flight Maintenance

    NASA Image and Video Library

    2003-10-10

    In the Orbiter Processing Facility, David Sanborn and Rick Cady, with United Space Alliance, check tiles on the underside of Endeavour. Tile check is part of routine maintenance and return to flight activities on the orbiter fleet.

  9. Contract Service for School Maintenance

    ERIC Educational Resources Information Center

    Modern Schools, 1976

    1976-01-01

    Preventive maintenance can extend useful equipment life in a school building and keep systems running more efficiently. Points to consider before selecting a comprehensive energy management package are listed. (Author/MLF)

  10. Avionics Maintenance Technology Program Standards.

    ERIC Educational Resources Information Center

    Georgia Univ., Athens. Dept. of Vocational Education.

    This publication contains statewide standards for the avionics maintenance technology program in Georgia. The standards are divided into the following categories: foundations, diploma/degree (philosophy, purpose, goals, program objectives, availability, evaluation); admissions, diploma/degree (admission requirements, provisional admission…

  11. Managing Custodial and Maintenance Staffs.

    ERIC Educational Resources Information Center

    Fickes, Michael

    2001-01-01

    Presents some basic maintenance management techniques that can help schools meet their budgets, preserve staffing levels, meet productivity needs, and sustain quality services. Tips for staff recruitment, training, and retention are explored. (GR)

  12. Maintenance and Monitoring of BMPS

    EPA Science Inventory

    Two best management practice (BMP) sites in the Staten Island Bluebelt in Richmond Creek Watershed are Richmond Creek 5 (RC-5) and Richmond Creek 4 (RC-4). This presentation includes site description, briefing of initial monitoring activity, representative maintenance activity, ...

  13. The Magic of Lighting Maintenance

    ERIC Educational Resources Information Center

    Modern Schools, 1973

    1973-01-01

    A well-planned lighting maintenance program can provide more light for your money, better appearance of the lighting system, lower overall cost of light, and fewer accidents and better visibility for users of the school. (Author)

  14. Genetic variation and its maintenance

    SciTech Connect

    Roberts, D.F.; De Stefano, G.F.

    1986-01-01

    This book contains several papers divided among three sections. The section titles are: Genetic Diversity--Its Dimensions; Genetic Diversity--Its Origin and Maintenance; and Genetic Diversity--Applications and Problems of Complex Characters.

  15. How to Get a Maintenance Program Underway

    ERIC Educational Resources Information Center

    Lundy, Lyndall L.

    1975-01-01

    The article describes the development of a comprehensive maintenance program for the school shop. A general maintenance management outline provides direction for planning, execution, and evaluation. (MW)

  16. How to Get a Maintenance Program Underway

    ERIC Educational Resources Information Center

    Lundy, Lyndall L.

    1975-01-01

    The article describes the development of a comprehensive maintenance program for the school shop. A general maintenance management outline provides direction for planning, execution, and evaluation. (MW)

  17. Software Tools for Software Maintenance

    DTIC Science & Technology

    1988-10-01

    Amisteant" project was commmioned to study the problems of software maintenance and to investigate the concept of bringing together a combinationof loosely...integrated tools that could improve the productivity o maintenance programmers and increase the reliability of modified programs. One area of study has...that can aid in understanding the program and modifying it. Background work for study in this area included in examination of existing software tools

  18. Dune Grass Fertilization and Maintenance

    DTIC Science & Technology

    1982-12-01

    plants often suffer, thereafter, from neglect. PURPOSE: To recommend a fertilization and maintenance program to sustain healthy dune grasses in coastal...TYPE 3. DATES COVERED 00-00-1982 to 00-00-1982 4. TITLE AND SUBTITLE Dune Grass Fertilization and Maintenance 5a. CONTRACT NUMBER 5b. GRANT...TO APPLY: Apply a granular or pelletized agricultural type fertilizer . It need not be a costly, slow- release material. A.t\\!OUNT OF FERTILIZER TO

  19. Membrane trafficking in neuronal maintenance and degeneration.

    PubMed

    Wang, Dong; Chan, Chih-Chiang; Cherry, Smita; Hiesinger, P Robin

    2013-08-01

    Defects in membrane trafficking and degradation are hallmarks of most, and maybe all, neurodegenerative disorders. Such defects typically result in the accumulation of undegraded proteins due to aberrant endosomal sorting, lysosomal degradation, or autophagy. The genetic or environmental cause of a specific disease may directly affect these membrane trafficking processes. Alternatively, changes in intracellular sorting and degradation can occur as cellular responses of degenerating neurons to unrelated primary defects such as insoluble protein aggregates or other neurotoxic insults. Importantly, altered membrane trafficking may contribute to the pathogenesis or indeed protect the neuron. The observation of dramatic changes to membrane trafficking thus comes with the challenging need to distinguish pathological from protective alterations. Here, we will review our current knowledge about the protective and destructive roles of membrane trafficking in neuronal maintenance and degeneration. In particular, we will first focus on the question of what type of membrane trafficking keeps healthy neurons alive in the first place. Next, we will discuss what alterations of membrane trafficking are known to occur in Alzheimer's disease and other tauopathies, Parkinson's disease, polyQ diseases, peripheral neuropathies, and lysosomal storage disorders. Combining the maintenance and degeneration viewpoints may yield insight into how to distinguish when membrane trafficking functions protectively or contributes to degeneration.

  20. 14 CFR 145.217 - Contract maintenance.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... product following contract maintenance, preventive maintenance, or alterations. ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Contract maintenance. 145.217 Section 145...) SCHOOLS AND OTHER CERTIFICATED AGENCIES REPAIR STATIONS Operating Rules § 145.217 Contract maintenance....

  1. 14 CFR 91.405 - Maintenance required.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance, and Alterations § 91.405 Maintenance required. Each owner or operator of an aircraft— (a) Shall have... 14 Aeronautics and Space 2 2012-01-01 2012-01-01 false Maintenance required. 91.405 Section...

  2. 14 CFR 91.405 - Maintenance required.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance, and Alterations § 91.405 Maintenance required. Each owner or operator of an aircraft— (a) Shall have... 14 Aeronautics and Space 2 2014-01-01 2014-01-01 false Maintenance required. 91.405 Section...

  3. 14 CFR 91.405 - Maintenance required.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance, and Alterations § 91.405 Maintenance required. Each owner or operator of an aircraft— (a) Shall have... 14 Aeronautics and Space 2 2013-01-01 2013-01-01 false Maintenance required. 91.405 Section...

  4. 46 CFR 122.702 - Maintenance.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Maintenance. 122.702 Section 122.702 Shipping COAST..., Maintenance, and Inspection of Lifesaving Equipment § 122.702 Maintenance. (a) The manufacturer's instructions for onboard maintenance of survival craft, rescue boats, and launching appliances, manufactured on...

  5. 18 CFR 367.81 - Maintenance.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 18 Conservation of Power and Water Resources 1 2011-04-01 2011-04-01 false Maintenance. 367.81... Expense Instructions § 367.81 Maintenance. (a) The cost of maintenance chargeable to the various operating... maintenance work. A list of work operations applicable generally to service company property is included...

  6. 23 CFR 633.208 - Maintenance.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 23 Highways 1 2011-04-01 2011-04-01 false Maintenance. 633.208 Section 633.208 Highways FEDERAL... PROVISIONS Federal-Aid Contracts (Appalachian Contracts) § 633.208 Maintenance. Maintenance of all highway... responsibility of the State. The State may arrange for maintenance of such roads or portions thereof,...

  7. 46 CFR 185.702 - Maintenance.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Maintenance. 185.702 Section 185.702 Shipping COAST...) OPERATIONS Operational Readiness, Maintenance, and Inspection of Lifesaving Equipment § 185.702 Maintenance. (a) The manufacturer's instructions for onboard maintenance of survival craft, rescue boats,...

  8. 22 CFR 120.38 - Maintenance levels.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 22 Foreign Relations 1 2014-04-01 2014-04-01 false Maintenance levels. 120.38 Section 120.38 Foreign Relations DEPARTMENT OF STATE INTERNATIONAL TRAFFIC IN ARMS REGULATIONS PURPOSE AND DEFINITIONS § 120.38 Maintenance levels. (a) Organizational-level maintenance (or basic-level maintenance) is...

  9. 46 CFR 185.702 - Maintenance.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Maintenance. 185.702 Section 185.702 Shipping COAST...) OPERATIONS Operational Readiness, Maintenance, and Inspection of Lifesaving Equipment § 185.702 Maintenance. (a) The manufacturer's instructions for onboard maintenance of survival craft, rescue boats, and...

  10. 14 CFR 91.405 - Maintenance required.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Maintenance required. 91.405 Section 91.405... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance, and Alterations § 91.405 Maintenance required. Each owner or operator of an aircraft— (a) Shall have...

  11. 23 CFR 633.208 - Maintenance.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 23 Highways 1 2010-04-01 2010-04-01 false Maintenance. 633.208 Section 633.208 Highways FEDERAL... PROVISIONS Federal-Aid Contracts (Appalachian Contracts) § 633.208 Maintenance. Maintenance of all highway... responsibility of the State. The State may arrange for maintenance of such roads or portions thereof, by...

  12. 14 CFR 91.417 - Maintenance records.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Maintenance records. 91.417 Section 91.417... AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance, and Alterations § 91.417 Maintenance records. (a) Except for work performed in accordance with §§ 91...

  13. 18 CFR 367.81 - Maintenance.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Maintenance. 367.81... Expense Instructions § 367.81 Maintenance. (a) The cost of maintenance chargeable to the various operating... maintenance work. A list of work operations applicable generally to service company property is included in...

  14. 46 CFR 122.702 - Maintenance.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Maintenance. 122.702 Section 122.702 Shipping COAST..., Maintenance, and Inspection of Lifesaving Equipment § 122.702 Maintenance. (a) The manufacturer's instructions for onboard maintenance of survival craft, rescue boats, and launching appliances, manufactured on or...

  15. Fast Flux Test Facility (FFTF) maintenance provisions

    SciTech Connect

    Marshall, J.L.

    1981-05-01

    The Fast Flux Test Facility (FFTF) was designed with maintainability as a primary parameter, and facilities and provisions were designed into the plant to accommodate the maintenance function. This paper describes the FFTF and its systems. Special maintenance equipment and facilities for performing maintenance on radioactive components are discussed. Maintenance provisions designed into the plant to enhance maintainability are also described.

  16. TPX remote maintenance and shielding

    SciTech Connect

    Rennich, M.J.; Nelson, B.E.

    1994-11-01

    The Tokamak Physics Experiment (TPX) machine design incorporates comprehensive planning for efficient and safe component maintenance. Three programmatic decisions have been made to insure the successful implementation of this objective. First, the tokamak incorporates radiation shielding to reduce activation of components and limit the dose rate to personnel working on the outside of the machine. This allows most of the ex-vessel equipment to be maintained through conventional {open_quotes}hands-on{close_quotes} procedures. Second, to the maximum extent possible, low activation materials will be used inside the shielding volume. This resulted in the selection of Titanium (Ti-6Al-4V) for the vacuum vessel and Plasma Facing Components (PFC) structures. The third decision stipulated that the primary in-vessel components will be replaced or repaired via remote maintenance tools specifically provided for the task. The component designers have been given the responsibility of incorporating maintenance design and for proving the maintainability of the design concepts in full-scale mockup tests prior to the initiation of final fabrication. Remote maintenance of the TPX machine is facilitated by general purpose tools provided by a special purpose design team. Major tools will include an in-vessel transporter, a vessel transfer system and a large component transfer container. In addition, tools such as manipulators and remotely operable impact wrenches will be made available to the component designers by this group. Maintenance systems will also provide the necessary controls for this equipment.

  17. TPX remote maintenance and shielding

    SciTech Connect

    Rennich, M.J.; Nelson, B.E.

    1994-09-01

    The Tokamak Physics Experiment machine design incorporates comprehensive planning for efficient and safe component maintenance. Three programmatic decisions have been made to insure the successful implementation of this objective. First, the tokamak incorporates radiation shielding to reduce activation of components and limit the dose rate to personnel working on the outside of the machine. This allows most of the ex-vessel equipment to be maintained through conventional ``hands-on`` procedures. Second, to the maximum extent possible, low activation materials will be used inside the shielding volume. This resulted in the selection of Titanium (Ti-6Al-4V) for the vacuum vessel and PFC structures. The third decision stipulated that the primary in-vessel components will be replaced or repaired via remote maintenance tools specifically provided for the task. The component designers have been given the responsibility of incorporating maintenance design and for proving the maintainability of the design concepts in full-scale mockup tests prior to the initiation of final fabrication. Remote maintenance of the TPX machine is facilitated by general purpose tools provided by a special purpose design team. Major tools will include an in-vessel transporter, a vessel transfer system and a large component transfer container. In addition, tools such as manipulators and remotely operable impact wrenches will be made available to the component designers by this group. Maintenance systems will also provide the necessary controls for this equipment.

  18. Iron production maintenance effectiveness system

    SciTech Connect

    Augstman, J.J.

    1996-12-31

    In 1989, an internal study in the Coke and Iron Maintenance Department identified the opportunities available to increase production, by decreasing unscheduled maintenance delays from 4.6%. A five year front loaded plan was developed, and presented to the company president. The plan required an initial investment of $1.4 million and a conservative break-even point was calculated to be 2.5 years. Due to budget restraints, it would have to be self-funded, i.e., generate additional production or savings, to pay for the program. The program began in 1991 at number 2 coke plant and the blast furnaces. This paper will describe the Iron Production Maintenance Effectiveness System (ME), which began with the mechanical and pipefitting trades.

  19. Streamflow Necessary for Environmental Maintenance

    NASA Astrophysics Data System (ADS)

    Whiting, Peter J.

    In the last decades, insights from the fields of ecology, geomorphology, and hydrology have been applied to the question of the streamflows necessary for environmental maintenance. For instance, determining the streamflow needed for spawning by salmon or trout requires ascertaining how much water, for how long, and at what time it will be needed? And what flows are necessary for the sustenance of streamside vegetation? Answers to these and similar questions have been sought to minimize environmental degradation in the development or relicensing of water projects, in restoring riverine ecosystems, and in balancing multiple uses for limited water resources. In this contribution, the varieties of environmental maintenance flows applied to rivers are described, as are their fundamental principles. These environmental maintenance flows include flows to maintain aesthetics and recreation, streambed sediment size and its mobility, the channel, its features and continuity, and the floodplain, its wetness regime, and riparian vegetation.

  20. Compliance improvement in periodontal maintenance.

    PubMed

    de Carvalho, Verônica Franco; Okuda, Osmar Shizuo; Bernardo, Carlos Cheque; Pannuti, Cláudio Mendes; Georgetti, Marco Antonio Paupério; De Micheli, Giorgio; Pustiglioni, Francisco Emílio

    2010-01-01

    The aim of this study was to assess the influence of efforts applied to modify the patients' behavior towards periodontal maintenance. Patients were classified into three groups: Complete Compliance (participation in all visits), Irregular Compliance (irregular participation, one or more missing appointments), and Noncompliance (abandoned or never returned to the program). Complete compliers received usual procedures of the maintenance visit. The irregular compliers and non-compliers received usual procedures and strategies such as reminding next visit, informing patients on both periodontal disease and importance of maintenance, motivating the patient who showed an improvement in compliance. Thus, 137 patients were observed for 12 months. The degree of compliance has increased significantly during this period (p=0.001). No association was detected between age or gender and compliance degree. The results have shown that the intervention applied had a favorable influence on the patients' compliance.

  1. NASA Critical Facilities Maintenance Assessment

    NASA Technical Reports Server (NTRS)

    Oberhettinger, David J.

    2006-01-01

    Critical Facilities Maintenance Assessment (CFMA) was first implemented by NASA following the March 2000 overtest of the High Energy Solar Spectroscopic Imager (HESSI) spacecraft. A sine burst dynamic test using a 40 year old shaker failed. Mechanical binding/slippage of the slip table imparted 10 times the planned force to the test article. There was major structural damage to HESSI. The mechanical "health" of the shaker had not been assessed and tracked to assure the test equipment was in good working order. Similar incidents have occurred at NASA facilities due to inadequate maintenance (e.g., rainwater from a leaky roof contaminated an assembly facility that housed a spacecraft). The HESSI incident alerted NASA to the urgent need to identify inadequacies in ground facility readiness and maintenance practices. The consequences of failures of ground facilities that service these NASA systems are severe due to the high unit value of NASA products.

  2. Development of preventive maintenance procedures.

    PubMed

    Ben-Zvi, S

    1984-01-01

    A large number of in-house preventive maintenance (PM) programs, which call for varying degrees of thoroughness in the checkout of patient care instrumentation, are currently in existence throughout the country. This paper discusses the types of preventive maintenance, or PM procedures, which can be used by a clinical engineering department; the rationale for drafting different types of PM procedures in-house; and some long-term considerations affecting hospital-based inspection programs. Three types of PM procedures are described and compared: general checks; generic procedures; and, specific procedures. An outline is provided for writing a PM procedure; and, a call is made for a national guideline for writing preventive maintenance procedures.

  3. NASA Critical Facilities Maintenance Assessment

    NASA Technical Reports Server (NTRS)

    Oberhettinger, David J.

    2006-01-01

    Critical Facilities Maintenance Assessment (CFMA) was first implemented by NASA following the March 2000 overtest of the High Energy Solar Spectroscopic Imager (HESSI) spacecraft. A sine burst dynamic test using a 40 year old shaker failed. Mechanical binding/slippage of the slip table imparted 10 times the planned force to the test article. There was major structural damage to HESSI. The mechanical "health" of the shaker had not been assessed and tracked to assure the test equipment was in good working order. Similar incidents have occurred at NASA facilities due to inadequate maintenance (e.g., rainwater from a leaky roof contaminated an assembly facility that housed a spacecraft). The HESSI incident alerted NASA to the urgent need to identify inadequacies in ground facility readiness and maintenance practices. The consequences of failures of ground facilities that service these NASA systems are severe due to the high unit value of NASA products.

  4. Advanced servomanipulator remote maintenance demonstration

    SciTech Connect

    Ray, T.L.; Bradley, E.C.

    1989-03-01

    The advanced servomanipulator (ASM) is a dual-arm, force-reflecting, master/slave servomanipulator that was designed for remote maintenance applications and is digitally controlled. The ASM is installed in the Maintenance Systems Test Area (MSTA) of the Fuel Recycle Division at the Oak Ridge National Laboratory. The unique aspect of ASM is that the slave arms were designed to be remotely maintainable, using a similar remote manipulator system, to maximize availability and minimize downtime. This test report describes the results of the maintenance testing conducted on ASM. Demonstration of the ability to maintain ASM remotely is an important precursor to the ultimate application of ASM in a totally remote facility. The approach taken in the design of ASM was for the manipulator slave arms to be composed of modules capable of being removed and replaced by another manipulator system of similar capabilities. The ASM incorporates gear and torque tube drives with drive couplings that facilitate remote maintenance. Although the use of special fixtures is normally discouraged for remote maintenance, special fixtures were required for this demonstration due to the complex nature of the slave arms. This test was performed to demonstrate that the ASM slave arms could be completely disassembled and reassembled remotely. Maintenance of ASM was successfully demonstrated using the M-2 servomanipulator and special fixtures. The entire disassembly process took about 4 h, and the assembly took about 3 1/2 h. Although there were some problems, in general, the arm modules were adequately designed for remote removal and replacement. Recommendations, which are documented in this report, have been made for improvements. 5 refs., 22 figs., 1 tab.

  5. Thermography instruments for predictive maintenance

    SciTech Connect

    Palko, E.

    1993-08-12

    Thermography (infrared imaging, or IR scanning) is not only the most versatile predictive maintenance technology available today; it is, in general, the most cost-effective. Plant engineering can apply a virtually unlimited variety of predictive maintenance instruments, but all are restricted regarding the types of existing and incipient problems they can detect. Inplant applications of thermography, however, are truly limited only by the extent of the plant engineer's imagination. Here are ways that thermography can be used to fight downtime in plants, and factors to consider when selecting the best instrument for particular circumstances.

  6. Maintenance as a corporate strategy.

    PubMed

    Clarke, D

    2000-11-01

    As we move towards the 21st century, those Manufacturing Industries that survive, will have changed their "efficiency" focus away from the conventional wisdom of "efficient individual tasks", to focus on the "effectiveness of a total manufacturing process". This seemingly insignificant change of emphasis, (from task to process) must include "the integration of process plant maintenance into the production process". Manufacturing companies, which do not include an effective plant maintenance strategy into their business strategy, will not survive the business environment of the 21st Century.

  7. A computerized hospital maintenance system.

    PubMed

    Kresch, E; Katz, P; Schwartz, H; Hamarman, H

    1985-01-01

    The Biomedical Instrumentation Department at Thomas Jefferson University Hospital maintains most of the clinical equipment owned by the hospital and provides support to six other hospitals, as well. In order to document these services, a computerized support system has been developed. This system maintains the inventory of equipment, documents the occurrence of repair and preventive maintenance procedures, generates lists of items due for maintenance and inspection, and prints reports and summaries of all activities performed by department staff. The system was designed for ease of use and requires a minimum of training for personnel who use it.

  8. Space Station maintenance concept study

    NASA Technical Reports Server (NTRS)

    Nelson, Eric E.

    1988-01-01

    The relationships among NASA Space Station operational constraints and logistical requirements are presently investigated. The concepts studied locate organizational, intermediate, and depot maintenance at the Space Station, at the Kennedy Space Center (KSC), and at a depot remote from the KSC. Measures of reliability, maintainability, and availability were selected; a life-cycle study was then conducted to ascertain the optimum Space Station system maintenance concept. The results obtained indicate that orbital replacement unit MTBFs should not be less than 36,000 hours.

  9. 14 CFR 125.245 - Organization required to perform maintenance, preventive maintenance, and alteration.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... GOVERNING PERSONS ON BOARD SUCH AIRCRAFT Maintenance § 125.245 Organization required to perform maintenance, preventive maintenance, and alteration. The certificate holder must ensure that each person with whom it... maintenance, preventive maintenance, and alteration. 125.245 Section 125.245 Aeronautics and Space FEDERAL...

  10. 14 CFR 125.245 - Organization required to perform maintenance, preventive maintenance, and alteration.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... GOVERNING PERSONS ON BOARD SUCH AIRCRAFT Maintenance § 125.245 Organization required to perform maintenance, preventive maintenance, and alteration. The certificate holder must ensure that each person with whom it... maintenance, preventive maintenance, and alteration. 125.245 Section 125.245 Aeronautics and Space FEDERAL...

  11. 14 CFR 125.245 - Organization required to perform maintenance, preventive maintenance, and alteration.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... maintenance, preventive maintenance, and alteration. 125.245 Section 125.245 Aeronautics and Space FEDERAL..., preventive maintenance, and alteration. The certificate holder must ensure that each person with whom it arranges for the performance of maintenance, preventive maintenance, alteration, or required...

  12. 14 CFR 91.407 - Operation after maintenance, preventive maintenance, rebuilding, or alteration.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 2 2012-01-01 2012-01-01 false Operation after maintenance, preventive... OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance, and Alterations § 91.407 Operation after maintenance, preventive maintenance, rebuilding, or alteration. (a) No person may operate any aircraft...

  13. 14 CFR 91.1425 - CAMP: Maintenance, preventive maintenance, and alteration programs.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 2 2013-01-01 2013-01-01 false CAMP: Maintenance, preventive maintenance... RULES Fractional Ownership Operations Program Management § 91.1425 CAMP: Maintenance, preventive... have an inspection program and a program covering other maintenance, preventive maintenance,...

  14. 14 CFR 91.407 - Operation after maintenance, preventive maintenance, rebuilding, or alteration.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 2 2013-01-01 2013-01-01 false Operation after maintenance, preventive... OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance, and Alterations § 91.407 Operation after maintenance, preventive maintenance, rebuilding, or alteration. (a) No person may operate any aircraft...

  15. 14 CFR 125.245 - Organization required to perform maintenance, preventive maintenance, and alteration.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... maintenance, preventive maintenance, and alteration. 125.245 Section 125.245 Aeronautics and Space FEDERAL..., preventive maintenance, and alteration. The certificate holder must ensure that each person with whom it arranges for the performance of maintenance, preventive maintenance, alteration, or required...

  16. 14 CFR 91.1425 - CAMP: Maintenance, preventive maintenance, and alteration programs.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 2 2014-01-01 2014-01-01 false CAMP: Maintenance, preventive maintenance... RULES Fractional Ownership Operations Program Management § 91.1425 CAMP: Maintenance, preventive... have an inspection program and a program covering other maintenance, preventive maintenance,...

  17. 14 CFR 91.1425 - CAMP: Maintenance, preventive maintenance, and alteration programs.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 2 2012-01-01 2012-01-01 false CAMP: Maintenance, preventive maintenance... RULES Fractional Ownership Operations Program Management § 91.1425 CAMP: Maintenance, preventive... have an inspection program and a program covering other maintenance, preventive maintenance,...

  18. 14 CFR 145.205 - Maintenance, preventive maintenance, and alterations performed for certificate holders under...

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Maintenance, preventive maintenance, and...) SCHOOLS AND OTHER CERTIFICATED AGENCIES REPAIR STATIONS Operating Rules § 145.205 Maintenance, preventive.... (a) A certificated repair station that performs maintenance, preventive maintenance, or...

  19. 14 CFR 125.245 - Organization required to perform maintenance, preventive maintenance, and alteration.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... maintenance, preventive maintenance, and alteration. 125.245 Section 125.245 Aeronautics and Space FEDERAL..., preventive maintenance, and alteration. The certificate holder must ensure that each person with whom it arranges for the performance of maintenance, preventive maintenance, alteration, or required...

  20. 14 CFR 145.205 - Maintenance, preventive maintenance, and alterations performed for certificate holders under...

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Maintenance, preventive maintenance, and...) SCHOOLS AND OTHER CERTIFICATED AGENCIES REPAIR STATIONS Operating Rules § 145.205 Maintenance, preventive.... (a) A certificated repair station that performs maintenance, preventive maintenance, or...

  1. 14 CFR 145.205 - Maintenance, preventive maintenance, and alterations performed for certificate holders under...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Maintenance, preventive maintenance, and...) SCHOOLS AND OTHER CERTIFICATED AGENCIES REPAIR STATIONS Operating Rules § 145.205 Maintenance, preventive.... (a) A certificated repair station that performs maintenance, preventive maintenance, or...

  2. 14 CFR 91.407 - Operation after maintenance, preventive maintenance, rebuilding, or alteration.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Operation after maintenance, preventive... OPERATING AND FLIGHT RULES Maintenance, Preventive Maintenance, and Alterations § 91.407 Operation after maintenance, preventive maintenance, rebuilding, or alteration. (a) No person may operate any aircraft that...

  3. Epigenetic regulation of memory formation and maintenance.

    PubMed

    Zovkic, Iva B; Guzman-Karlsson, Mikael C; Sweatt, J David

    2013-01-15

    Understanding the cellular and molecular mechanisms underlying the formation and maintenance of memories is a central goal of the neuroscience community. It is well regarded that an organism's ability to lastingly adapt its behavior in response to a transient environmental stimulus relies on the central nervous system's capability for structural and functional plasticity. This plasticity is dependent on a well-regulated program of neurotransmitter release, post-synaptic receptor activation, intracellular signaling cascades, gene transcription, and subsequent protein synthesis. In the last decade, epigenetic markers like DNA methylation and post-translational modifications of histone tails have emerged as important regulators of the memory process. Their ability to regulate gene transcription dynamically in response to neuronal activation supports the consolidation of long-term memory. Furthermore, the persistent and self-propagating nature of these mechanisms, particularly DNA methylation, suggests a molecular mechanism for memory maintenance. In this review, we will examine the evidence that supports a role of epigenetic mechanisms in learning and memory. In doing so, we hope to emphasize (1) the widespread involvement of these mechanisms across different behavioral paradigms and distinct brain regions, (2) the temporal and genetic specificity of these mechanisms in response to upstream signaling cascades, and (3) the functional outcome these mechanisms may have on structural and functional plasticity. Finally, we consider the future directions of neuroepigenetic research as it relates to neuronal storage of information.

  4. Epigenetic regulation of memory formation and maintenance

    PubMed Central

    Zovkic, Iva B.; Guzman-Karlsson, Mikael C.; Sweatt, J. David

    2013-01-01

    Understanding the cellular and molecular mechanisms underlying the formation and maintenance of memories is a central goal of the neuroscience community. It is well regarded that an organism's ability to lastingly adapt its behavior in response to a transient environmental stimulus relies on the central nervous system's capability for structural and functional plasticity. This plasticity is dependent on a well-regulated program of neurotransmitter release, post-synaptic receptor activation, intracellular signaling cascades, gene transcription, and subsequent protein synthesis. In the last decade, epigenetic markers like DNA methylation and post-translational modifications of histone tails have emerged as important regulators of the memory process. Their ability to regulate gene transcription dynamically in response to neuronal activation supports the consolidation of long-term memory. Furthermore, the persistent and self-propagating nature of these mechanisms, particularly DNA methylation, suggests a molecular mechanism for memory maintenance. In this review, we will examine the evidence that supports a role of epigenetic mechanisms in learning and memory. In doing so, we hope to emphasize (1) the widespread involvement of these mechanisms across different behavioral paradigms and distinct brain regions, (2) the temporal and genetic specificity of these mechanisms in response to upstream signaling cascades, and (3) the functional outcome these mechanisms may have on structural and functional plasticity. Finally, we consider the future directions of neuroepigenetic research as it relates to neuronal storage of information. PMID:23322554

  5. Checklist for School Maintenance Surveys.

    ERIC Educational Resources Information Center

    Maryland State Dept. of Education, Baltimore.

    This document is a simple 34-category checklist to be used by technicians conducting maintenance surveys. Categories includes: roadways & parking lots; site appearance; site utilities; exterior appearance; playground equipment; exterior structural conditions; gutters and downspouts; windows and caulking; sidewalks; entryways and exit doors;…

  6. Building Maintenance. Student Learning Guide.

    ERIC Educational Resources Information Center

    Palm Beach County Board of Public Instruction, West Palm Beach, FL.

    This student learning guide contains one module for completing a course in building maintenance. It is designed especially for use in secondary schools in Palm Beach County, Florida. The module covers one task, and consists of a purpose, performance objective, enabling objectives, learning activities keyed to resources, information sheets, student…

  7. Locker Room Maintenance Made Easy.

    ERIC Educational Resources Information Center

    Theel, James

    1998-01-01

    Provides examples on ways to make locker room maintenance easier and their use more student-friendly. Improvements include use of indoor-outdoor carpeting with numerous floor drains to cut mildew buildup, adequate ventilation to reduce musty smells, better hot water management, ceramic tiles to reduce water-damage repair and painting needs, and…

  8. Circassian Language Maintenance in Jordan

    ERIC Educational Resources Information Center

    Rannut, Ulle

    2009-01-01

    The central goal of this research is to explore the language policy aspects in Jordan by focusing on the Circassian language maintenance issues and to provide measures for language revitalisation in the current demographic, linguistic and political situation. Research is based on multiple sources of information, but primarily on the empirical data…

  9. Drive alignment pays maintenance dividends

    SciTech Connect

    Fedder, R.

    2008-12-15

    Proper alignment of the motor and gear drive on conveying and processing equipment will result in longer bearing and coupling life, along with lower maintenance costs. Selecting an alignment free drive package instead of a traditional foot mounted drive and motor is a major advancement toward these goals. 4 photos.

  10. Maintenance Downtime August 14, 2013

    Atmospheric Science Data Center

    2013-08-14

    Date(s):  Wednesday, August 14, 2013 Time:  7:00 am - 1:00 pm EST Event Impact:  Due to scheduled maintenance on Wednesday, August 14, 2013: • Connection the the ASDC website will be unavailable from 8 ...

  11. Afrikaans Language Maintenance in Australia

    ERIC Educational Resources Information Center

    Hatoss, Aniko; Starks, Donna; van Rensburg, Henriette Janse

    2011-01-01

    Changes in the political climate in the home country have resulted in the emigration of South Africans to English speaking countries such as Britain, Canada, Australia and New Zealand. Despite the scale of movement of the South African population, language maintenance in these diasporic contexts has received little consideration. This paper…

  12. Integration, Resegregation and Integration Maintenance.

    ERIC Educational Resources Information Center

    Peterman, William A.

    Recent increases in black migration to the suburbs and the continuing existence of discrimination in housing have emphasized the issues of integration and resegregation in suburban municipalities. To prevent resegregation, many integrated municipalities have adopted integration maintenance measures such as efforts to inform people that racial…

  13. Methods to much improve maintenance.

    PubMed

    Lam, K C

    2004-11-01

    The benefits to be gained from the application of both value engineering (VE) and quality systems--e.g. quality assurance (QA), and total quality management (TQM)--to the management of building services maintenance have yet to be realised by the building services industry. These concepts are particularly applicable to complex hospital engineering services.

  14. Building and Home Maintenance Services.

    ERIC Educational Resources Information Center

    Michigan State Univ., East Lansing. Coll. of Agriculture and Natural Resources Education Inst.

    This task-based curriculum guide for building and home maintenance services is intended to help the teacher develop a classroom management system where students learn by doing. Introductory materials include a Dictionary of Occupational Titles job code and title sheet, a career ladder, a matrix relating duty/task numbers to job titles, and a task…

  15. Advanced Tools for Software Maintenance.

    DTIC Science & Technology

    1982-12-01

    Old Applications ...... 118 11.3.3 Training People to Use New Tools ......... 119 Appendix A. Ada Style Guidelines . . . . . . . . . . . . . 121...and application -specific programming techniques and methods. - The Intelligent Editor provides facilities for manipulating programs at several...are applicable today or in the near future. In identifying tools and techniques, this study focused on one aspect of the maintenance problem

  16. Maintenance Sessions Prolong Cigarette Abstinence.

    ERIC Educational Resources Information Center

    Brandon, Thomas H.; And Others

    Recent smoking treatment programs have shifted emphasis from initial cessation rates to long-term abstinence, with aversion therapy and coping response training having had the most success. A smoking cessation treatment consisting of rapid smoking and behavioral counseling was supplemented with two maintenance treatments. After completing the…

  17. The Benefits of Health Maintenance.

    ERIC Educational Resources Information Center

    Rosenstein, Alan H.

    1987-01-01

    The article focuses on the merits of a comprehensive, medically-oriented health maintenance/risk assessment program, and suggests that such conditions as heart disease, cancer, and arteriosclerosis can be prevented or postponed through proper nutrition, weight control, exercise, smoking cessation, and stress management. (Author/CB)

  18. Avionics Maintenance Technology Program Guide.

    ERIC Educational Resources Information Center

    Georgia Univ., Athens. Dept. of Vocational Education.

    This program guide presents the avionics maintenance technology curriculum for technical institutes in Georgia. The general information section contains the following for both the diploma program and the associate degree program: purpose and objectives; program description, including admissions, typical job titles, and accreditation and…

  19. School Facilities Maintenance and Operations.

    ERIC Educational Resources Information Center

    Association of School Business Officials of the United States and Canada, Park Ridge, IL. Research Corp.

    This publication presents a series of field-proven school energy conservation, management, maintenance, and operations practices and ideas. Also included are the names, addresses, and phone numbers of individuals and organizations to contact for more detailed information. The brief summaries are grouped into six sections. "Planning and…

  20. New Directions in Maintenance Simulation.

    ERIC Educational Resources Information Center

    Miller, Gary G.

    A two-phase effort was conducted to design and evaluate a maintenance simulator which incorporated state-of-the-art information in simulation and instructional technology. The particular equipment selected to be simulated was the 6883 Convert/Flight Controls Test Station. Phase I included a generalized block diagram of the computer-trainer, the…