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Sample records for body component protein

  1. Drosophila SMN complex proteins Gemin2, Gemin3, and Gemin5 are components of U bodies

    Energy Technology Data Exchange (ETDEWEB)

    Cauchi, Ruben J.; Sanchez-Pulido, Luis [MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX (United Kingdom); Liu, Ji-Long, E-mail: jilong.liu@dpag.ox.ac.uk [MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX (United Kingdom)

    2010-08-15

    Uridine-rich small nuclear ribonucleoproteins (U snRNPs) play key roles in pre-mRNA processing in the nucleus. The assembly of most U snRNPs takes place in the cytoplasm and is facilitated by the survival motor neuron (SMN) complex. Discrete cytoplasmic RNA granules called U bodies have been proposed to be specific sites for snRNP assembly because they contain U snRNPs and SMN. U bodies invariably associate with P bodies, which are involved in mRNA decay and translational control. However, it remains unknown whether other SMN complex proteins also localise to U bodies. In Drosophila there are four SMN complex proteins, namely SMN, Gemin2/CG10419, Gemin3 and Gemin5/Rigor mortis. Drosophila Gemin3 was originally identified as the Drosophila orthologue of human and yeast Dhh1, a component of P bodies. Through an in silico analysis of the DEAD-box RNA helicases we confirmed that Gemin3 is the bona fide Drosophila orthologue of vertebrate Gemin3 whereas the Drosophila orthologue of Dhh1 is Me31B. We then made use of the Drosophila egg chamber as a model system to study the subcellular distribution of the Gemin proteins as well as Me31B. Our cytological investigations show that Gemin2, Gemin3 and Gemin5 colocalise with SMN in U bodies. Although they are excluded from P bodies, as components of U bodies, Gemin2, Gemin3 and Gemin5 are consistently found associated with P bodies, wherein Me31B resides. In addition to a role in snRNP biogenesis, SMN complexes residing in U bodies may also be involved in mRNP assembly and/or transport.

  2. The adenovirus E4 11 k protein binds and relocalizes the cytoplasmic P-body component Ddx6 to aggresomes

    International Nuclear Information System (INIS)

    The adenovirus E4 11 k protein, product of E4 ORF3, is required in infection for processes including normal accumulation of viral late mRNAs. 11 k restructures both the nucleus and cytoplasm of infected cells by relocalizing specific host cell target proteins, most strikingly components of nuclear PML oncogenic domains. It is likely that in many cases relocalization inactivates target proteins to produce 11 k's effects, although the mechanism and targets for stimulation of late mRNA accumulation is unknown. We have identified a new set of proteins relocalized by 11 k: at least five protein components of cytoplasmic mRNA processing bodies (p-bodies) are found in 11 k-induced cytoplasmic aggresomes, sites where proteins are inactivated or destroyed. One of these p-body proteins, RNA helicase Ddx6, binds 11 k, suggesting a mechanism for relocalization. Because p-bodies are sites for mRNA degradation, their modification by 11 k may provide an explanation for the role of 11 k in viral late mRNA accumulation.

  3. Effects of pH on protein components of extracted oil bodies from diverse plant seeds and endogenous protease-induced oleosin hydrolysis.

    Science.gov (United States)

    Zhao, Luping; Chen, Yeming; Chen, Yajing; Kong, Xiangzhen; Hua, Yufei

    2016-06-01

    Plant seeds are used to extract oil bodies for diverse applications, but oil bodies extracted at different pH values exhibit different properties. Jicama, sunflower, peanut, castor bean, rapeseed, and sesame were selected to examine the effects of pH (6.5-11.0) on the protein components of oil bodies and the oleosin hydrolysis in pH 6.5-extracted oil bodies. In addition to oleosins, many extrinsic proteins (globulins, 2S albumin, and enzymes) were present in pH 6.5-extracted oil bodies. Globulins were mostly removed at pH 8.0, whereas 2S albumins were removed at pH 11.0. At pH 11.0, highly purified oil bodies were obtained from jicama, sunflower, peanut, and sesame, whereas lipase remained in the castor bean oil bodies and many enzymes in the rapeseed oil bodies. Endogenous protease-induced hydrolysis of oleosins occurred in all selected plant seeds. Oleosins with larger sizes were hydrolysed more quickly than oleosins with smaller sizes in each plant seed. PMID:26830569

  4. Body composition analysis: Cellular level modeling of body component ratios

    OpenAIRE

    Z. Wang; Heymsfield, S. B.; PI-SUNYER, F.X.; Gallagher, D.; PIERSON, R.N.

    2008-01-01

    During the past two decades, a major outgrowth of efforts by our research group at St. Luke’s-Roosevelt Hospital is the development of body composition models that include cellular level models, models based on body component ratios, total body potassium models, multi-component models, and resting energy expenditure-body composition models. This review summarizes these models with emphasis on component ratios that we believe are fundamental to understanding human body composition during growt...

  5. Measurements of body protein for clinical investigation

    International Nuclear Information System (INIS)

    Body protein (nitrogen) is determined by bilaterally irradiating the body with neutrons using Pu-Be sources and measuring the resultant 10.8 MeV gamma rays from the reaction 14N(n,8) 15N. In the authors lab the whole body can be scanned or separate segments of the body can be measured independently. A nitrogen index has been developed based on body size and is used as a predictor of normal total body nitrogen (TBN). They have found that TBN, when normalized to body size in this way, provides a reliable index of protein status which cannot be accurately determined by body weight, anthropometry, or body potassium measurements. Changes in body composition with age were studied by measuring the composition of 56 healthy female volunteers aged 20-80. Measurements were made for K(40K), Ca and N. It was shown that protein and bone mineral decrease with age but that this is not reflected in K or anthropometry measurements. Results of other studies to be presented include: body protein measurements pre and post TPN (total parenteral nutrition), nutritional status of patients on long term CAPD (continuous ambulatory peritoneal dialysis) and changes in body composition as a result of TPN in patients with small cell lung cancer receiving chemotherapy. Clinical results show that indirect measurements of body protein based on weight, potassium, or anthropometry, do not give an accurate measure of body protein. For an accurate measurement, direct measurement of body protein is necessary

  6. Rape embryogenesis VI. Formation of protein bodies

    OpenAIRE

    Teresa Tykarska

    2014-01-01

    The storage protein synthesis starts in Brassica napus var. Górczański embryo at the final embryogenesis stage, i.e. in green seeds. Storage protein accumulate in selected zones adjacent to big vacuoles. These vacuoles, as well as surrounding protein zones, are subject to fragmentation. Young aleuron grains originate. They grow occupying sites of declining vacuoles. In mature rape embryo two kinds of protein bodies occur: aleuron grains, well-stainable with protein-specific dyes, and myrosin ...

  7. Structure and Composition of Protein Bodies from Wild-Type and High-Lysine Barley Endosperm

    DEFF Research Database (Denmark)

    Ingversen, J.

    1975-01-01

    granular component. Particles with the same structure were present in the protein body preparation from the mutant, where, however, the granular component was the most prominent. Amino-acid composition and SDS-polyacrylamide gel electrophoresis of the proteins from the protein body preparation revealed......-polyacrylamidegel electrophoresis. Sections through pellets of isolated protein bodies from both the mutant and the wild type revealed protein body structures corresponding with those observed in sections through the intact starchy endosperms. The majority of the wild-type protein bodies was homogeneous spheres accompanied with a...

  8. Pasiflora proteins are novel core components of the septate junction.

    Science.gov (United States)

    Deligiannaki, Myrto; Casper, Abbie L; Jung, Christophe; Gaul, Ulrike

    2015-09-01

    Epithelial sheets play essential roles as selective barriers insulating the body from the environment and establishing distinct chemical compartments within it. In invertebrate epithelia, septate junctions (SJs) consist of large multi-protein complexes that localize at the apicolateral membrane and mediate barrier function. Here, we report the identification of two novel SJ components, Pasiflora1 and Pasiflora2, through a genome-wide glial RNAi screen in Drosophila. Pasiflora mutants show permeable blood-brain and tracheal barriers, overelongated tracheal tubes and mislocalization of SJ proteins. Consistent with the observed phenotypes, the genes are co-expressed in embryonic epithelia and glia and are required cell-autonomously to exert their function. Pasiflora1 and Pasiflora2 belong to a previously uncharacterized family of tetraspan membrane proteins conserved across the protostome-deuterostome divide. Both proteins localize at SJs and their apicolateral membrane accumulation depends on other complex components. In fluorescence recovery after photobleaching experiments we demonstrate that pasiflora proteins are core SJ components as they are required for complex formation and exhibit restricted mobility within the membrane of wild-type epithelial cells, but rapid diffusion in cells with disrupted SJs. Taken together, our results show that Pasiflora1 and Pasiflora2 are novel integral components of the SJ and implicate a new family of tetraspan proteins in the function of these ancient and crucial cell junctions. PMID:26329602

  9. Abundant lipid and protein components of drusen.

    Directory of Open Access Journals (Sweden)

    Lan Wang

    Full Text Available BACKGROUND: Drusen are extracellular lesions characteristic of aging and age-related maculopathy, a major retinal disease of the elderly. We determined the relative proportions of lipids and proteins in drusen capped with retinal pigment epithelium (RPE and in RPE isolated from non-macular regions of 36 human retinas with grossly normal maculas obtained <6 hr after death. METHODOLOGY/PRINCIPAL FINDINGS: Druse pellets were examined by light and electron microscopy. Component proteins were extracted using novel methods for preserved tissues, separated, subjected to tryptic digestion and LC-MS(MS(2 analysis using an ion trap mass spectrometer, and identified with reference to databases. Lipid classes were separated using thin layer chromatography and quantified by densitometry. Major druse components were esterified cholesterol (EC, phosphatidylcholine (PC, and protein (37.5+/-13.7, 36.9+/-12.9, and 43.0+/-11.5 ng/druse, respectively. Lipid-containing particles (median diameter, 77 nm occupied 37-44% of druse volume. Major proteins include vitronectin, complement component 9, apoE, and clusterin, previously seen in drusen, and ATP synthase subunit beta, scavenger receptor B2, and retinol dehydrogenase 5, previously seen in RPE. Drusen and RPE had similar protein profiles, with higher intensities and greater variability in drusen. C8, part of the complement membrane attack complex, was localized in drusen by immunofluorescence. CONCLUSIONS/SIGNIFICANCE: At least 40% of druse content is comprised by lipids dominated by EC and PC, 2 components that are potentially accounted for by just one pathway, the secretion of lipoproteins by RPE. Manipulating genes encoding apolipoprotein pathways would be a fruitful approach to producing drusen with high EC content in laboratory animals. Therapies that directly mitigate drusen should prepare for the substantial volume of neutral lipids. The catalog of major druse proteins is nearing completion.

  10. A STUDY ON BODY COMPOSITION, BODY COMPONENTS AND SOMATOTYPE CHARACTERISTICS OF SOCCER PLAYERS

    OpenAIRE

    Kürkçü, Recep; Hazar, Fatih; Özdağ, Selçuk

    2009-01-01

    The aim of this study is to study to determine and comparison the body composition, body components and somatotype characteristics of young soccer players (Young Soccer Team of Sport Club of Muğla University) with other national and international soccer players. Subjects were eighteen pubescent soccer players (age, 13.22y) of a team playing in regional soccer league. Skinfolds (biceps, triceps, back, suprailiac, abdominal, leg, thigh), diameters (femur and humerus biconduler), circumferences ...

  11. Structural components of the nuclear body in nuclei of Allium cepa cells

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Nuclear bodies have long been noted in interphase nuclei of plant cells,but their structural component,origin and function are still unclear by now.The present work showed in onion cells the nuclear bodies appeared as a spherical structure about 0.3 to 0.8 μm in diameter.They possibly were formed in nucleolus and subsequently released,and entered into nucleoplasm.Observation through cytochemical staining method at the ultrastructural level confirmed that nuclear bodies consisted of ribonucleoproteins (RNPs) and silver-stainable proteins.Immunocytochemical results revealed that nuclear bodies contained no DNA and ribosomal gene transcription factor (UBF).Based on these data,we suggested that nuclear bodies are not related to the ribosome or other gene transcription activities,instead they may act as subnuclear structures for RNPs transport from nucleolus to cytoplasm,and may also be involved in splicing of pre-mRNAs.

  12. Variance components for body weight in Japanese quails (Coturnix japonica

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    RO Resende

    2005-03-01

    Full Text Available The objective of this study was to estimate the variance components for body weight in Japanese quails by Bayesian procedures. The body weight at hatch (BWH and at 7 (BW07, 14 (BW14, 21 (BW21 and 28 days of age (BW28 of 3,520 quails was recorded from August 2001 to June 2002. A multiple-trait animal model with additive genetic, maternal environment and residual effects was implemented by Gibbs sampling methodology. A single Gibbs sampling with 80,000 rounds was generated by the program MTGSAM (Multiple Trait Gibbs Sampling in Animal Model. Normal and inverted Wishart distributions were used as prior distributions for the random effects and the variance components, respectively. Variance components were estimated based on the 500 samples that were left after elimination of 30,000 rounds in the burn-in period and 100 rounds of each thinning interval. The posterior means of additive genetic variance components were 0.15; 4.18; 14.62; 27.18 and 32.68; the posterior means of maternal environment variance components were 0.23; 1.29; 2.76; 4.12 and 5.16; and the posterior means of residual variance components were 0.084; 6.43; 22.66; 31.21 and 30.85, at hatch, 7, 14, 21 and 28 days old, respectively. The posterior means of heritability were 0.33; 0.35; 0.36; 0.43 and 0.47 at hatch, 7, 14, 21 and 28 days old, respectively. These results indicate that heritability increased with age. On the other hand, after hatch there was a marked reduction in the maternal environment variance proportion of the phenotypic variance, whose estimates were 0.50; 0.11; 0.07; 0.07 and 0.08 for BWH, BW07, BW14, BW21 and BW28, respectively. The genetic correlation between weights at different ages was high, except for those estimates between BWH and weight at other ages. Changes in body weight of quails can be efficiently achieved by selection.

  13. Skin Inqjuries Reduce Survival and Modulate Corticosterone, C-Reactive Protein, Complement Component 3, IgM, and Prostaglandin E2 after Whole-Body Reactor-Produced Mixed Field (n + γ-Photons Irradiation

    Directory of Open Access Journals (Sweden)

    Juliann G. Kiang

    2013-01-01

    Full Text Available Skin injuries such as wounds or burns following whole-body γ-irradiation (radiation combined injury (RCI increase mortality more than whole-body γ-irradiation alone. Wound-induced decreases in survival after irradiation are triggered by sustained activation of inducible nitric oxide synthase pathways, persistent alteration of cytokine homeostasis, and increased susceptibility to systemic bacterial infection. Among these factors, radiation-induced increases in interleukin-6 (IL-6 concentrations in serum were amplified by skin wound trauma. Herein, the IL-6-induced stress proteins including C-reactive protein (CRP, complement 3 (C3, immunoglobulin M (IgM, and prostaglandin E2 (PGE2 were evaluated after skin injuries given following a mixed radiation environment that might be found after a nuclear incident. In this report, mice received 3 Gy of reactor-produced mixed field (n+γ-photons radiations at 0.38 Gy/min followed by nonlethal skin wounding or burning. Both wounds and burns reduced survival and increased CRP, C3, and PGE2 in serum after radiation. Decreased IgM production along with an early rise in corticosterone followed by a subsequent decrease was noted for each RCI situation. These results suggest that RCI-induced alterations of corticosterone, CRP, C3, IgM, and PGE2 cause homeostatic imbalance and may contribute to reduced survival. Agents inhibiting these responses may prove to be therapeutic for RCI and improve related survival.

  14. A STUDY ON BODY COMPOSITION, BODY COMPONENTS AND SOMATOTYPE CHARACTERISTICS OF SOCCER PLAYERS

    Directory of Open Access Journals (Sweden)

    Recep KÜRKÇÜ

    2009-08-01

    Full Text Available The aim of this study is to study to determine and comparison the body composition, body components and somatotype characteristics of young soccer players (Young Soccer Team of Sport Club of Muğla University with other national and international soccer players. Subjects were eighteen pubescent soccer players (age, 13.22y of a team playing in regional soccer league. Skinfolds (biceps, triceps, back, suprailiac, abdominal, leg, thigh, diameters (femur and humerus biconduler, circumferences (biceps, thigh of the body and body fat parameters were measured. Somatotype characteristics were calculated and evaluated by Heat-Carter formula. Subjects’ measurements were as; height 158.44±10.42cm, body weight 47.65±8.38kg, skinfolds; biceps 5.75±1.54mm, triceps 10.61±2.93mm, back 7.30±1.59mm, suprailiac 7.00±2.04mm, abdominal 9.91±3.98mm, leg 13.52±4.76mm; diameters; femur biconduler 11.03±0.74cm; humerus biconduler 7.30±0.59cm; circumferences, biceps 22.76±3.11cm, thigh 32.84±3.33cm and body fat percentage 5.41±1.37 %, somatotype characteristics; Endomorph; 4.59±2.08, Mezomorph; 6.94±3.10, and Ecthomorph; 3.55±1.34. In related sports, physical fitness parameters including physical and anthropometric characteristics of athletes are very important in talent identification. Therefore, results of the present study could provide important data on selection of talented players in soccer and to the other related researches.

  15. In-service weld repairs of valve bodies and components

    International Nuclear Information System (INIS)

    The idea of performing welding and machining to valve bodies in line, has only come of age within the last five to seven years. Although many plants currently use this technology, many limit its use to light machine cuts, lapping or polishing of seats and/or gasket faces. It's no secret, over the past ten years technology has advanced rapidly. The technological resources available today, in this specialized industry, are practically incalculable. Suppliers are stocking fewer parts, thus generating longer lead times. As a result, the need to repair has overwhelmed the desire to replace. Technological advances in machine welding processes and specialized machine tooling have turned what was once considered science, into reality. Over the past few years these type services have greatly improved the quality of repairs and the time it takes to complete them. As in any new technology there are typically some concerns. They may range from, ''Did I get a sound repair or a Band-Aid fix?'' to ''Was it really cost effective?'' There are only a few requirements one needs to follow: (1) define the Work Scope ''detail''; (2) qualify a vendor; (3) perform mockup; (4) be ready for the unexpected. The idea behind this Paper is to show its readers that in line repairs require considerable planning, communication, teamwork, capable personnel, and state-of-the-art equipment. The advantage of an in line valve repair versus removing it from its in line location is simply, cost effectiveness. Occasionally, environmental factors may even prohibit such removal. Additionally, it is not unusual for valve replacement to become critical path during an outage. In summary, the author intends to elaborate on the full potential and benefits of in line repairs to valve bodies and related components

  16. Proteins Are the Body's Worker Molecules

    Science.gov (United States)

    ... to Top Parts of Some Proteins Fold Into Corkscrews A mix of alpha helices and beta sheets. ... shapes. Where a protein chain curves into a corkscrew, that section is called an alpha helix. Where ...

  17. Skin Injuries Reduce Survival and Modulate Corticosterone, C-Reactive Protein, Complement Component 3, IgM, and Prostaglandin E 2 after Whole-Body Reactor-Produced Mixed Field (n + γ-Photons) Irradiation

    OpenAIRE

    Kiang, Juliann G; Ledney, G. David

    2013-01-01

    Skin injuries such as wounds or burns following whole-body γ-irradiation (radiation combined injury (RCI)) increase mortality more than whole-body γ-irradiation alone. Wound-induced decreases in survival after irradiation are triggered by sustained activation of inducible nitric oxide synthase pathways, persistent alteration of cytokine homeostasis, and increased susceptibility to systemic bacterial infection. Among these factors, radiation-induced increases in interleukin-6 (IL-6) concentrat...

  18. Skin Inqjuries Reduce Survival and Modulate Corticosterone, C-Reactive Protein, Complement Component 3, IgM, and Prostaglandin E2 after Whole-Body Reactor-Produced Mixed Field (n + γ-Photons) Irradiation

    OpenAIRE

    Kiang, Juliann G; G. David Ledney

    2013-01-01

    Skin injuries such as wounds or burns following whole-body γ-irradiation (radiation combined injury (RCI)) increase mortality more than whole-body γ-irradiation alone. Wound-induced decreases in survival after irradiation are triggered by sustained activation of inducible nitric oxide synthase pathways, persistent alteration of cytokine homeostasis, and increased susceptibility to systemic bacterial infection. Among these factors, radiation-induced increases in interleukin-6 (IL-6) concentrat...

  19. A high protein diet upregulated whole-body protein turnover during energy deficit

    Science.gov (United States)

    The effects of higher protein diets and sustained energy deficit (ED) on whole-body protein turnover (WBPTO) are not well described. This study examined whether dietary protein level influences whole-body protein breakdown (Ra), non-oxidative leucine disposal (NOLD), and oxidation (Ox) during ED. ...

  20. Effect of Various Protein Sources on Body Weight Development

    DEFF Research Database (Denmark)

    Rønnevik, Alexander Krokedal

    Background: Due to the increasing prevalence of obesity, finding effective dietary strategies for weight loss and weight maintenance is of great interest. High protein diets are reported to protect against diet-induced obesity, however less is known about how different protein sources affect body...... consumption of lean meat in Western background diets was only evident with free access to the diets, most likely due to differences in body composition. We purpose that the beneficial effects of lean seafood consumption in relation to body weight regulation may be due to an enrichment of the amino acids...... weight regulation. We aimed to investigate how various protein sources influenced body weight development and glucose metabolism by feeding obesity prone male C57/BL6 mice various protein sources in different background diets. Results: In high fat/high sucrose diets (HF/HS), high fat/high protein diets...

  1. The dynamics of protein body formation in developing wheat grain.

    Science.gov (United States)

    Moore, Katie L; Tosi, Paola; Palmer, Richard; Hawkesford, Malcolm J; Grovenor, Chris R M; Shewry, Peter R

    2016-09-01

    Wheat is a major source of protein in the diets of humans and livestock but we know little about the mechanisms that determine the patterns of protein synthesis in the developing endosperm. We have used a combination of enrichment with (15) N glutamine and NanoSIMS imaging to establish that the substrate required for protein synthesis is transported radially from its point of entrance in the endosperm cavity across the starchy endosperm tissues, before becoming concentrated in the cells immediately below the aleurone layer. This transport occurs continuously during grain development but may be slower in the later stages. Although older starchy endosperm cells tend to contain larger protein deposits formed by the fusion of small protein bodies, small highly enriched protein bodies may also be present in the same cells. This shows a continuous process of protein body initiation, in both older and younger starchy endosperm cells and in all regions of the tissue. Immunolabeling with specific antibodies shows that the patterns of enrichment are not related to the contents of gluten proteins in the protein bodies. In addition to providing new information on the dynamics of protein deposition, the study demonstrates the wider utility of NanoSIMS and isotope labelling for studying complex developmental processes in plant tissues. PMID:26898533

  2. Protein structure similarity from principle component correlation analysis

    Directory of Open Access Journals (Sweden)

    Chou James

    2006-01-01

    Full Text Available Abstract Background Owing to rapid expansion of protein structure databases in recent years, methods of structure comparison are becoming increasingly effective and important in revealing novel information on functional properties of proteins and their roles in the grand scheme of evolutionary biology. Currently, the structural similarity between two proteins is measured by the root-mean-square-deviation (RMSD in their best-superimposed atomic coordinates. RMSD is the golden rule of measuring structural similarity when the structures are nearly identical; it, however, fails to detect the higher order topological similarities in proteins evolved into different shapes. We propose new algorithms for extracting geometrical invariants of proteins that can be effectively used to identify homologous protein structures or topologies in order to quantify both close and remote structural similarities. Results We measure structural similarity between proteins by correlating the principle components of their secondary structure interaction matrix. In our approach, the Principle Component Correlation (PCC analysis, a symmetric interaction matrix for a protein structure is constructed with relationship parameters between secondary elements that can take the form of distance, orientation, or other relevant structural invariants. When using a distance-based construction in the presence or absence of encoded N to C terminal sense, there are strong correlations between the principle components of interaction matrices of structurally or topologically similar proteins. Conclusion The PCC method is extensively tested for protein structures that belong to the same topological class but are significantly different by RMSD measure. The PCC analysis can also differentiate proteins having similar shapes but different topological arrangements. Additionally, we demonstrate that when using two independently defined interaction matrices, comparison of their maximum

  3. Where do Protein Bodies of Cereal Seeds Come From?

    Science.gov (United States)

    Pedrazzini, Emanuela; Mainieri, Davide; Marrano, Claudia A.; Vitale, Alessandro

    2016-01-01

    Protein bodies of cereal seeds consist of ordered, largely insoluble heteropolymers formed by prolamin storage proteins within the endoplasmic reticulum (ER) of developing endosperm cells. Often these structures are permanently unable to traffic along the secretory pathway, thus representing a unique example for the use of the ER as a protein storage compartment. In recent years, marked progress has been made in understanding what is needed to make a protein body and in formulating hypotheses on how protein body formation might have evolved as an efficient mechanism to store large amounts of protein during seed development, as opposed to the much more common system of seed storage protein accumulation in vacuoles. The major key evolutionary events that have generated prolamins appear to have been insertions or deletions that have disrupted the conformation of the eight-cysteine motif, a protein folding motif common to many proteins with different functions and locations along the secretory pathway, and, alternatively, the fusion between the eight-cysteine motif and domains containing additional cysteine residues. PMID:27540384

  4. Pulsed Joining Of Body-In-White Components

    Energy Technology Data Exchange (ETDEWEB)

    Bonnen, John [Ford Motor Company, Dearborn, MI (United States)

    2014-09-30

    The objective of this project was to develop cost efficient high quality pulsed welding (PW) technology for joining 6xxx Aluminum and High Strength Steel (with tensile strength above 580MPa) components enabling broad usage of hydroformed parts and leading to substantial weight reduction of cars and trucks to reduce US demand on petroleum, lower carbon emissions and energy expenditures. In general, pulsed welding is a form of impact welding where two dissimilar metal pieces are joined by accelerating one to velocities exceeding 300m/s at which point the first piece strikes the second and forms a weld. In this work, two methods were used to accelerate the flyer material: Electro-Magnetic (EM) pulse and Electro-Hydraulic (EH) pulse launching. The advantage of pulsed welding techniques is that welds can be formed between two materials that cannot otherwise be welded: high strength aluminum and high strength steel. The technical objectives of the project included: 1) developing cost affordable production feasible tooling design for PW of 6xxx aluminum to High Strength steel with strengths above 580MPa; 2) demonstrating that fabricated joints can exceed the required service load strength initially at the coupon level and then at the component level; 3) developing fundamental understanding of the mechanisms of joint formation and conditions leading to formation of high quality PW joint; and 4) creating a numerical model predicting the tooling and electric discharge parameters necessary for the joint formation and that satisfy the targeted strength parameters. The project successfully developed: 1) EM and EH pulsed welds between high strength aluminum with tensile strengths exceeding 240MPa and steels exceeding 580MPa; 2) pulsed welds of extrusions with strengths exceeding project requirements; 3) EM and EH flyer launch models and 4) weld interface formation models. However, the grant holder, Ford Motor Company, could see no path to commercialization and the work was

  5. Plasma surfactant protein D levels and the relation to body mass index in a chinese population

    DEFF Research Database (Denmark)

    Zhao, X M; Wu, Y P; Wei, R; Cai, H X; Tornoe, I; Han, J J; Wang, Y; de Groot, P G; Holmskov, U; Xia, Z L; Sørensen, Grith Lykke

    2007-01-01

    significant effect of age, and (iii) a significant inverse association between serum SP-D and body mass index (BMI) (P = 0.012). The data indicate that racial differences in SP-D expression exist as the median plasma SP-D in the Chinese population was approximately two times lower than the median serum SP......Surfactant protein D (SP-D) is a member of the collectin family and is an important component of the pulmonary innate host defence. The protein has a widespread distribution in the human body and is present in multiple epithelia, in endothelium and in blood. Various studies have looked at the...

  6. Leishmanial protein kinases phosphorylate components of the complement system.

    OpenAIRE

    Hermoso, T; Fishelson, Z; Becker, S I; Hirschberg, K.; Jaffe, C. L.

    1991-01-01

    Externally oriented protein kinases are present on the plasma membrane of the human parasite, Leishmania. Since activation of complement plays an important role in the survival of these parasites, we examined the ability of protein kinases from Leishmania major to phosphorylate components of the human complement system. The leishmanial protein kinase-1 (LPK-1) isolated from promastigotes of L. major was able to phosphorylate purified human C3, C5 and C9. Only the alpha-chain of C3 and C5 was ...

  7. Characterization of relative growth of empty body and carcass components for bulls from a five-breed diallel.

    Science.gov (United States)

    Baker, J F; Bryson, W L; Sanders, J O; Dahm, P F; Cartwright, T C; Ellis, W C; Long, C R

    1991-08-01

    Slaughter and carcass data were obtained on 197 bulls produced in a diallel involving Angus, Brahman, Hereford, Holstein and Jersey that were slaughtered at either 6, 9, 12, 15, 18, 24, or 30 mo of age. Bulls were given ad libitum access to a 72% TDN diet on an individual basis from 6 mo of age until slaughter. Empty body weight (EBWT) was determined as the sum of the weights of blood, hide, hard drop, soft drop (minus contents of the digestive tract), and carcass weight (CWT), which were recorded at slaughter. Carcass protein (CPROT) and fat (CFAT) were based on weights and chemical analyses of lean and fat tissue and bone of the carcass. Empty body protein (EBPROT) and fat (EBFAT) were based on weights and chemical estimates of the components of the empty body. Growth of EBWT, EBPROT, EBFAT, CWT, CPROT, and CFAT relative to either live weight (LWT), EBWT, or CWT were investigated using the allometric equation. Breed-type differences existed (P less than .01) for the growth of EBWT relative to LWT. Comparisons of general combining abilities revealed that Angus, Hereford, and Jersey generally had lower maturing rates of EBWT relative to LWT and that Brahman and Holstein had higher maturing rates. Across breed-type, relative growth rates indicated that fat and protein were later-maturing components relative to LWT, EBWT, or CWT, which implies that other components mature relatively earlier. Relative maturing rates of components studied were not important in explaining differences in body composition that have been previously reported for these breed-types. PMID:1894551

  8. Protein intake during hemodialysis maintains a positive whole body protein balance in chronic hemodialysis patients

    NARCIS (Netherlands)

    Veeneman, JM; Kingma, HA; Boer, TS; Stellaard, F; De Jong, PE; Reijngoud, DJ; Huisman, RM

    2003-01-01

    Protein energy malnutrition is present in 18 to 56% of hemodialysis patients. Because hemodialysis has been regarded as a catabolic event, we studied whether consumption of a protein- and energy-nriched meal improves the whole body protein balance during dialysis in chronic hemodialysis (CHD) patien

  9. Identification of the RNAs for Transcription Factor Mitf as a Component of the Balbiani Body

    Institute of Scientific and Technical Information of China (English)

    Mingyou Li; Yongming Yuan; Yunhan Hong

    2013-01-01

    Balbiani body (BB) is a large distinctive organelle aggregate uniquely present in developing oocytes of diverse animal species.BB is thought as a stage-specific structure that resembles germ plasm,the cytoplasmic organelle of germ cells.The role and function of BB have remained speculative because of a highly dynamic structure and a lack of genetic and molocular data.BB has been found to contain proteins and RNAs,none of them-except the zebrafish foxH1 RNA,is or encodes a transcription factor.Here we report in the fish medaka (Oryzias latipes) that RNAs encoding microphthalmia-associated transcription factor (Mitf) are prominent components of the BB.By fluorescence in siru hybridization on ovarian section,we revealed that the transcripts of both mitfl and mitf2 genes concentrated in the BB,in which they co-localized with the dazl RNA,a definitive BB marker highly conserved in vertebrates.Therefore,the mitfproduct may play dual roles in germ gene transcription and BB formation and/or function in this organism.Our data provide the second evidence that the RNA of a transcription factor can be a prominent component of the BB in a vertebrate.

  10. Protein cages, rings and tubes: useful components of future nanodevices?

    Directory of Open Access Journals (Sweden)

    Jonathan G Heddle

    2008-11-01

    Full Text Available Jonathan G HeddleGlobal Edge Institute, Tokyo Institute of Technology, Nagatsuda, Midori-ku, Yokohama Kanagawa, JapanAbstract: There is a great deal of interest in the possibility that complex nanoscale devices can be designed and engineered. Such devices will lead to the development of new materials, electronics and smart drugs. Producing complex nanoscale devices, however will present many challenges and the components of such devices will require a number of special features. Devices will be engineered to incorporate desired functionalities but, because of the difficulties of controlling matter precisely at the nanoscale with current technology, the nanodevice components must self-assemble. In addition, nanocomponents that are to have wide applicability in various devices must have enough flexibility to integrate into a large number of potentially very different environments. These challenges are daunting and complex, and artificial nanodevices have not yet been constructed. However, the existence of nanomachines in nature in the form of proteins (eg, enzymes suggests that they will be possible to produce. As the material from which nature’s nanomachines are made, proteins seem ideal to form the basis of engineered components of such nanodevices. Initially, engineering projects may focus on building blocks such as rings, cages and tubes, examples of which exist in nature and may act as a useful start point for modification and further development. This review focuses on the recent research and possible future development of such protein building blocks.Keywords: bionanotechnology, protein engineering, nanomachine, building-blocks, synthetic biology

  11. Endogenous and exogenous components in the circadian variation of core body temperature in humans

    NARCIS (Netherlands)

    Hiddinga, AE; Beersma, DGM; VandenHoofdakker, RH

    1997-01-01

    Core body temperature is predominantly modulated by endogenous and exogenous components. In the present study we tested whether these two components can be reliably assessed in a protocol which lasts for only 120 h. In this so-called forced desynchrony protocol, 12 healthy male subjects (age 23.7 +/

  12. A giant protein associated with the anterior pole of a trypanosomatid cell body skeleton.

    Science.gov (United States)

    Baqui, M M; Takata, C S; Milder, R V; Pudles, J

    1996-07-01

    A megadalton protein was found to be a cytoskeleton component of the promastigote forms of the flagellate Phytomonas serpens. This protein migrated on sodium dodecyl sulfate polyacrylamide gel electrophoresis as a doublet of polypeptides with a molecular mass similar to muscle beta-connectin (titin) 2500-3000 kDa. A polyclonal antibody raised against this protein reacts, by immunoblot analysis, with Phytomonas serpens and two others Phytomonas species. In addition, the Phytomonas serpens protein was immunoprecipitated after being metabolically labeled with [35S]methionine. This antibody did not cross-react with the cytoskeletal proteins of Trypanosoma cruzi, Crithidia luciliae thermophila, Crithidia fasciculata and Leptomonas samueli or with beta-connectin (titin). Indirect immunofluorescence microscopy analysis revealed a punctate fluorescence staining at the anterior region of the parasite's body skeleton. Moreover, immunogold electron microscopy of cytoskeletal preparations and of thin sections of whole cells indicates that the giant protein appears to cap the anterior end of the cell body microtubules at the level of the junctional complex. We suggest that this giant protein may serve as a linker between the cell body skeleton and the flagellum membrane. PMID:8832208

  13. Proteomic characterisation of endoplasmic reticulum-derived protein bodies in tobacco leaves

    Directory of Open Access Journals (Sweden)

    Joseph Minu

    2012-03-01

    Full Text Available Abstract Background The N-terminal proline-rich domain (Zera of the maize storage protein γ-zein, is able to induce the formation of endoplasmic reticulum (ER-derived protein bodies (PBs when fused to proteins of interest. This encapsulation enables a recombinant fused protein to escape from degradation and facilitates its recovery from plant biomass by gradient purification. The aim of the present work was to evaluate if induced PBs encapsulate additional proteins jointly with the recombinant protein. The exhaustive analysis of protein composition of PBs is expected to facilitate a better understanding of PB formation and the optimization of recombinant protein purification approaches from these organelles. Results We analysed the proteome of PBs induced in Nicotiana benthamiana leaves by transient transformation with Zera fused to a fluorescent marker protein (DsRed. Intact PBs with their surrounding ER-membrane were isolated on iodixanol based density gradients and their integrity verified by confocal and electron microscopy. SDS-PAGE analysis of isolated PBs showed that Zera-DsRed accounted for around 85% of PB proteins in term of abundance. Differential extraction of PBs was performed for in-depth analysis of their proteome and structure. Besides Zera-DsRed, 195 additional proteins were identified including a broad range of proteins resident or trafficking through the ER and recruited within the Zera-DsRed polymer. Conclusions This study indicates that Zera-protein fusion is still the major protein component of the new formed organelle in tobacco leaves. The analysis also reveals the presence of an unexpected diversity of proteins in PBs derived from both the insoluble Zera-DsRed polymer formation, including ER-resident and secretory proteins, and a secretory stress response induced most likely by the recombinant protein overloading. Knowledge of PBs protein composition is likely to be useful to optimize downstream purification of

  14. Principal component analysis based methodology to distinguish protein SERS spectra

    Science.gov (United States)

    Das, G.; Gentile, F.; Coluccio, M. L.; Perri, A. M.; Nicastri, A.; Mecarini, F.; Cojoc, G.; Candeloro, P.; Liberale, C.; De Angelis, F.; Di Fabrizio, E.

    2011-05-01

    Surface-enhanced Raman scattering (SERS) substrates were fabricated using electro-plating and e-beam lithography techniques. Nano-structures were obtained comprising regular arrays of gold nanoaggregates with a diameter of 80 nm and a mutual distance between the aggregates (gap) ranging from 10 to 30 nm. The nanopatterned SERS substrate enabled to have better control and reproducibility on the generation of plasmon polaritons (PPs). SERS measurements were performed for various proteins, namely bovine serum albumin (BSA), myoglobin, ferritin, lysozyme, RNase-B, α-casein, α-lactalbumin and trypsin. Principal component analysis (PCA) was used to organize and classify the proteins on the basis of their secondary structure. Cluster analysis proved that the error committed in the classification was of about 14%. In the paper, it was clearly shown that the combined use of SERS measurements and PCA analysis is effective in categorizing the proteins on the basis of secondary structure.

  15. Protein Bodies in Leaves Exchange Contents through the Endoplasmic Reticulum

    Science.gov (United States)

    Saberianfar, Reza; Sattarzadeh, Amirali; Joensuu, Jussi J.; Kohalmi, Susanne E.; Menassa, Rima

    2016-01-01

    Protein bodies (PBs) are organelles found in seeds whose main function is the storage of proteins that are used during germination for sustaining growth. PBs can also be induced to form in leaves when foreign proteins are produced at high levels in the endoplasmic reticulum (ER) and when fused to one of three tags: Zera®, elastin-like polypeptides (ELP), or hydrophobin-I (HFBI). In this study, we investigate the differences between ELP, HFBI and Zera PB formation, packing, and communication. Our results confirm the ER origin of all three fusion-tag-induced PBs. We show that secretory pathway proteins can be sequestered into all types of PBs but with different patterns, and that different fusion tags can target a specific protein to different PBs. Zera PBs are mobile and dependent on actomyosin motility similar to ELP and HFBI PBs. We show in vivo trafficking of proteins between PBs using GFP photoconversion. We also show that protein trafficking between ELP or HFBI PBs is faster and proteins travel further when compared to Zera PBs. Our results indicate that fusion-tag-induced PBs do not represent terminally stored cytosolic organelles, but that they form in, and remain part of the ER, and dynamically communicate with each other via the ER. We hypothesize that the previously documented PB mobility along the actin cytoskeleton is associated with ER movement rather than independent streaming of detached organelles. PMID:27242885

  16. Protein bodies in leaves exchange contents through the endoplasmic reticulum

    Directory of Open Access Journals (Sweden)

    Reza eSaberianfar

    2016-05-01

    Full Text Available Protein bodies (PBs are organelles found in seeds whose main function is the storage of proteins that are used during germination for sustaining growth. PBs can also be induced to form in leaves when foreign proteins are produced at high levels in the endoplasmic reticulum (ER and when fused to one of three tags: Zera®, elastin-like polypeptides (ELP, or hydrophobin-I (HFBI. In this study, we investigate the differences between ELP, HFBI and Zera PB formation, packing, and communication. Our results confirm the ER origin of all three fusion-tag-induced PBs. We show that secretory pathway proteins can be sequestered into all types of PBs but with different patterns, and that different fusion tags can target a specific protein to different PBs. Zera PBs are mobile and dependent on actomyosin motility similar to ELP and HFBI PBs. We show in vivo trafficking of proteins between PBs using GFP photoconversion. We also show that protein trafficking between ELP or HFBI PBs is faster and proteins travel further when compared to Zera PBs. Our results indicate that fusion-tag-induced PBs do not represent terminally stored cytosolic organelles, but that they form in, and remain part of the ER, and dynamically communicate with each other via the ER. We hypothesize that the previously documented PB mobility along the actin cytoskeleton is associated with ER movement rather than independent streaming of detached organelles.

  17. Total body protein in chronic diseases and in aging.

    Science.gov (United States)

    Hansen, R D; Raja, C; Allen, B J

    2000-05-01

    Substantial losses of total body protein (TBP) can occur in chronic diseases and in aging. Such losses impact negatively on immunity and quality of life, and on growth rates in children. Direct measurements of total body nitrogen (TBN) monitor the integrated changes in TBP over time and allow comparison with normal subjects. TBN assessment via neutron capture analysis is therefore the gold-standard method of TBP estimation, so that risk factors for protein deficit can be identified and patient management optimized. The nitrogen index (NI) can be used to predict prognostic outcome: an NI breast-cancer patients. These findings emphasize the central importance of adequate protein stores in recovery from disease or in maintaining quality of life. Aging appears to involve a gradual loss of TBP throughout adulthood. Cross-sectional data suggest that TBP declines curvilinearly with age, such that there is an accelerated decline after 65 years of age. However, longitudinal data are scarce, and little is known about the relative loss of visceral protein, as opposed to skeletal muscle protein. More clearly-defined data are essential if the effects of aging per se are to be separated from the effects of chronic disease. A further complication is the knowledge that physical activity also declines with age. Thus sarcopenia, the loss of skeletal muscle mass, could primarily result from disuse rather than aging. The economic impact of unsuccessful aging places a pressing need for multicompartment data in longitudinal study designs. PMID:10865769

  18. The interplay between components of the mitochondrial protein translocation motor studied using purified components.

    Science.gov (United States)

    Slutsky-Leiderman, Olga; Marom, Milit; Iosefson, Ohad; Levy, Ran; Maoz, Sharon; Azem, Abdussalam

    2007-11-23

    The final step of protein translocation across the mitochondrial inner membrane is mediated by a translocation motor composed of 1) the matrix-localized, ATP-hydrolyzing, 70-kDa heat shock protein mHsp70; 2) its anchor to the import channel, Tim44; 3) the nucleotide exchange factor Mge1; and 4) a J-domain-containing complex of co-chaperones, Tim14/Pam18-Tim16/Pam16. Despite its essential role in the biogenesis of mitochondria, the mechanism by which the translocation motor functions is still largely unknown. The goal of this work was to carry out a structure-function analysis of the mitochondrial translocation motor utilizing purified components, with an emphasis on the formation of the Tim44-mHsp70 complex. To this end, we purified Tim44 and monitored its interaction with other components of the motor using cross-linking with bifunctional reagents. The effects of nucleotides, the J-domain-containing components, and the P5 peptide (CALLSAPRR, representing part of the mitochondrial targeting signal of aspartate aminotransferase) on the formation of the translocation motor were examined. Our results show that only the peptide and nucleotides, but not J-domain-containing proteins, affect the Tim44-mHsp70 interaction. Additionally, binding of Tim44 to mHsp70 prevents the formation of a complex between the latter and Tim14/Pam18-Tim16/Pam16. Thus, mutually exclusive interactions between various components of the motor with mHsp70 regulate its functional cycle. The results are discussed in light of known models for the function of the mitochondrial translocation motor. PMID:17881357

  19. Turnover of whole body proteins and myofibrillar proteins in middle-aged active men

    International Nuclear Information System (INIS)

    Endurance-trained older men have a higher proportion of lean tissue and greater muscle cell oxidative capacity, reversing age-related trends and suggesting major changes in protein metabolism. In this study, protein turnover was determined in 6 middle-aged (52+/-1 yr) men who were well trained (VO2 max 55.2+/-5.0 ml O2/kg.min) and lean (body fat 18.9+/-2.8%, muscle mass 36.6+/-0.6%). The maintained habitual exercise while consuming 0.6, 0.9 or 1.2 g protein/kg.day for 10-day periods. N flux was measured from 15N in urea after oral 15N-glycine administration. Myofibrillar protein breakdown was estimated from urinary 3-methyl-histidine. Dietary protein had no effect on turnover rates, even when N balance was negative. Whole body protein synthesis was 3.60+/-0.12 g/kg.day and breakdown was 3.40+/-0.14 g/kg.day for all N intakes. Whole body protein flux, synthesis and breakdown were similar to values reported for sedentary young (SY) or sedentary old (SO) men on comparable diets. 3-me-his (3.67+/-0.14 μmol/kg.day) was similar to values reported for SY but higher (p<0.01) than for SO. Myofibrillar protein breakdown per unit muscle mass (185+/-7 μmol 3-me-his/g creatinine) was higher (p<0.01) than for SY or SO. In active middle-aged men, myofibrillar proteins may account for a greater proportion of whole body protein turnover, despite an age-related reduction in muscle mass

  20. A principal components approach to parent-to-newborn body composition associations in South India

    Directory of Open Access Journals (Sweden)

    Hill Jacqueline C

    2009-02-01

    Full Text Available Abstract Background Size at birth is influenced by environmental factors, like maternal nutrition and parity, and by genes. Birth weight is a composite measure, encompassing bone, fat and lean mass. These may have different determinants. The main purpose of this paper was to use anthropometry and principal components analysis (PCA to describe maternal and newborn body composition, and associations between them, in an Indian population. We also compared maternal and paternal measurements (body mass index (BMI and height as predictors of newborn body composition. Methods Weight, height, head and mid-arm circumferences, skinfold thicknesses and external pelvic diameters were measured at 30 ± 2 weeks gestation in 571 pregnant women attending the antenatal clinic of the Holdsworth Memorial Hospital, Mysore, India. Paternal height and weight were also measured. At birth, detailed neonatal anthropometry was performed. Unrotated and varimax rotated PCA was applied to the maternal and neonatal measurements. Results Rotated PCA reduced maternal measurements to 4 independent components (fat, pelvis, height and muscle and neonatal measurements to 3 components (trunk+head, fat, and leg length. An SD increase in maternal fat was associated with a 0.16 SD increase (β in neonatal fat (p Conclusion Principal components analysis is a useful method to describe neonatal body composition and its determinants. Newborn adiposity is related to maternal nutritional status and parity, while newborn length is genetically determined. Further research is needed to understand mechanisms linking maternal pelvic size to fetal growth and the determinants and implications of the components (trunk v leg length of fetal skeletal growth.

  1. Variance Component Quantitative Trait Locus Analysis for Body Weight Traits in Purebred Korean Native Chicken.

    Science.gov (United States)

    Cahyadi, Muhammad; Park, Hee-Bok; Seo, Dong-Won; Jin, Shil; Choi, Nuri; Heo, Kang-Nyeong; Kang, Bo-Seok; Jo, Cheorun; Lee, Jun-Heon

    2016-01-01

    Quantitative trait locus (QTL) is a particular region of the genome containing one or more genes associated with economically important quantitative traits. This study was conducted to identify QTL regions for body weight and growth traits in purebred Korean native chicken (KNC). F1 samples (n = 595) were genotyped using 127 microsatellite markers and 8 single nucleotide polymorphisms that covered 2,616.1 centi Morgan (cM) of map length for 26 autosomal linkage groups. Body weight traits were measured every 2 weeks from hatch to 20 weeks of age. Weight of half carcass was also collected together with growth rate. A multipoint variance component linkage approach was used to identify QTLs for the body weight traits. Two significant QTLs for growth were identified on chicken chromosome 3 (GGA3) for growth 16 to18 weeks (logarithm of the odds [LOD] = 3.24, Nominal p value = 0.0001) and GGA4 for growth 6 to 8 weeks (LOD = 2.88, Nominal p value = 0.0003). Additionally, one significant QTL and three suggestive QTLs were detected for body weight traits in KNC; significant QTL for body weight at 4 weeks (LOD = 2.52, nominal p value = 0.0007) and suggestive QTL for 8 weeks (LOD = 1.96, Nominal p value = 0.0027) were detected on GGA4; QTLs were also detected for two different body weight traits: body weight at 16 weeks on GGA3 and body weight at 18 weeks on GGA19. Additionally, two suggestive QTLs for carcass weight were detected at 0 and 70 cM on GGA19. In conclusion, the current study identified several significant and suggestive QTLs that affect growth related traits in a unique resource pedigree in purebred KNC. This information will contribute to improving the body weight traits in native chicken breeds, especially for the Asian native chicken breeds. PMID:26732327

  2. Body image disturbance in binge eating disorder: a comparison of obese patients with and without binge eating disorder regarding the cognitive, behavioral and perceptual component of body image.

    Science.gov (United States)

    Lewer, Merle; Nasrawi, Nadia; Schroeder, Dorothea; Vocks, Silja

    2016-03-01

    Whereas the manifestation of body image disturbance in binge eating disorder (BED) has been intensively investigated concerning the cognitive-affective component, with regard to the behavioral and the perceptual components of body image disturbance in BED, research is limited and results are inconsistent. Therefore, the present study assessed body image disturbance in BED with respect to the different components of body image in a sample of obese females (n = 31) with BED compared to obese females without an eating disorder (n = 28). The Eating Disorder Inventory-2, the Eating Disorder Examination-Questionnaire, the Body Image Avoidance Questionnaire and the Body Checking Questionnaire as well as a Digital Photo Distortion Technique based on a picture of each participant taken under standardized conditions were employed. Using two-sample t tests, we found that the participants with BED displayed significantly greater impairments concerning the cognitive-affective component of body image than the control group. Concerning the behavioral component, participants with BED reported more body checking and avoidance behavior than the controls, but group differences failed to reach significance after the Bonferroni corrections. Regarding the perceptual component, a significant group difference was found for the perceived "ideal" figure, with the individuals suffering from BED displaying a greater wish for a slimmer ideal figure than the control group. These results support the assumption that body image disturbance is a relevant factor in BED, similar to other eating disorders. PMID:26178486

  3. Stable Isotope Labeling Strategy for Protein-Ligand Binding Analysis in Multi-Component Protein Mixtures

    Science.gov (United States)

    DeArmond, Patrick D.; West, Graham M.; Huang, Hai-Tsang; Fitzgerald, Michael C.

    2011-03-01

    Described here is a stable isotope labeling protocol that can be used with a chemical modification- and mass spectrometry-based protein-ligand binding assay for detecting and quantifying both the direct and indirect binding events that result from protein-ligand binding interactions. The protocol utilizes an H{2/16}O2 and H{2/18}O2 labeling strategy to evaluate the chemical denaturant dependence of methionine oxidation in proteins both in the presence and absence of a target ligand. The differential denaturant dependence to the oxidation reactions performed in the presence and absence of ligand provides a measure of the protein stability changes that occur as a result of direct interactions of proteins with the target ligand and/or as a result of indirect interactions involving other protein-ligand interactions that are either induced or disrupted by the ligand. The described protocol utilizes the 18O/16O ratio in the oxidized protein samples to quantify the ligand-induced protein stability changes. The ratio is determined using the isotopic distributions observed for the methionine-containing peptides used for protein identification in the LC-MS-based proteomics readout. The strategy is applied to a multi-component protein mixture in this proof-of-principle experiment, which was designed to evaluate the technique's ability to detect and quantify the direct binding interaction between cyclosporin A and cyclophilin A and to detect the indirect binding interaction between cyclosporin A and calcineurin (i.e., the protein-protein interaction between cyclophilin A and calcineurin that is induced by cyclosporin A binding to cyclophilin A).

  4. Nuclear Fragile X Mental Retardation Protein is localized to Cajal bodies.

    Directory of Open Access Journals (Sweden)

    Alain Y Dury

    2013-10-01

    Full Text Available Fragile X syndrome is caused by loss of function of a single gene encoding the Fragile X Mental Retardation Protein (FMRP. This RNA-binding protein, widely expressed in mammalian tissues, is particularly abundant in neurons and is a component of messenger ribonucleoprotein (mRNP complexes present within the translational apparatus. The absence of FMRP in neurons is believed to cause translation dysregulation and defects in mRNA transport essential for local protein synthesis and for synaptic development and maturation. A prevalent model posits that FMRP is a nucleocytoplasmic shuttling protein that transports its mRNA targets from the nucleus to the translation machinery. However, it is not known which of the multiple FMRP isoforms, resulting from the numerous alternatively spliced FMR1 transcripts variants, would be involved in such a process. Using a new generation of anti-FMRP antibodies and recombinant expression, we show here that the most commonly expressed human FMRP isoforms (ISO1 and 7 do not localize to the nucleus. Instead, specific FMRP isoforms 6 and 12 (ISO6 and 12, containing a novel C-terminal domain, were the only isoforms that localized to the nuclei in cultured human cells. These isoforms localized to specific p80-coilin and SMN positive structures that were identified as Cajal bodies. The Cajal body localization signal was confined to a 17 amino acid stretch in the C-terminus of human ISO6 and is lacking in a mouse Iso6 variant. As FMRP is an RNA-binding protein, its presence in Cajal bodies suggests additional functions in nuclear post-transcriptional RNA metabolism. Supporting this hypothesis, a missense mutation (I304N, known to alter the KH2-mediated RNA binding properties of FMRP, abolishes the localization of human FMRP ISO6 to Cajal bodies. These findings open unexplored avenues in search for new insights into the pathophysiology of Fragile X Syndrome.

  5. A connected component-based method for efficiently integrating multiscale $N$-body systems

    OpenAIRE

    Jänes, Jürgen; Pelupessy, Federico I.; Zwart, Simon F. Portegies

    2014-01-01

    We present a novel method for efficient direct integration of gravitational N-body systems with a large variation in characteristic time scales. The method is based on a recursive and adaptive partitioning of the system based on the connected components of the graph generated by the particle distribution combined with an interaction-specific time step criterion. It uses an explicit and approximately time-symmetric time step criterion, and conserves linear and angular momentum to machine preci...

  6. Casein and soya-bean protein have different effects on whole body protein turnover at the same nitrogen balance

    DEFF Research Database (Denmark)

    Nielsen, K; Kondrup, J; Elsner, Petteri;

    1994-01-01

    The present study examined whether different proteins have different effects on whole-body protein turnover in adult rats. The rats were either starved, given a protein-free but energy-sufficient diet (1 MJ/kg body weight (BW) per d) or a diet containing intact casein, hydrolysed casein, or...... recovered from urinary ammonia and urea during isotope steady state for measurement of protein synthesis and protein degradation. Compared with starvation the protein-free diet decreased N excretion by 75%, probably by increasing the rate of reutilization of amino acids from endogenous proteins for protein......-bean protein did not increase protein synthesis but decreased protein degradation by 35% compared with the protein-free diet. Compared with the hydrolysed soya-bean protein, intact casein resulted in 2.2- and 2.8-fold higher rates of protein synthesis and degradation respectively. These results are not easily...

  7. The P Body Protein Dcp1a Is Hyper-phosphorylated during Mitosis

    OpenAIRE

    Adva Aizer; Pinhas Kafri; Alon Kalo; Yaron Shav-Tal

    2013-01-01

    Processing bodies (PBs) are non-membranous cytoplasmic structures found in all eukaryotes. Many of their components such as the Dcp1 and Dcp2 proteins are highly conserved. Using live-cell imaging we found that PB structures disassembled as cells prepared for cell division, and then began to reassemble during the late stages of cytokinesis. During the cell cycle and as cells passed through S phase, PB numbers increased. However, there was no memory of PB numbers between mother and daughter ce...

  8. Calcium distribution in globoid crystals of cucurbita cotyledon protein bodies.

    Science.gov (United States)

    Lott, J N; Spitzer, E; Vollmer, C M

    1979-05-01

    Energy-dispersive x-ray analysis was used to investigate the location of globoid crystals with relatively high Ca levels within cotyledons of Cucurbita maxima, Cucurbita mixta, and Cucurbita andreana. The small globoid crystals in both upper and lower epidermal cells commonly contained Ca. Ca was present in globoid crystals of all provascular regions with the exception of the very small provascular regions of C. maxima. In C. maxima and C. mixta cotyledons, some cases were observed where Ca was found in the globoid crystals of the first layer of mesophyll cells surrounding the provascular region, but in general Ca was absent from globoid crystals of palisade and spongy mesophyll cells. In C. andreana, globoid crystals of palisade and spongy mesophyll cells commonly contained at least some Ca. Cell position and cell type are factors affecting the Ca content of globoid crystals in protein bodies. PMID:16660825

  9. Casein and soya-bean protein have different effects on whole body protein turnover at the same nitrogen balance

    DEFF Research Database (Denmark)

    Nielsen, K; Kondrup, J; Elsner, Petteri;

    1994-01-01

    The present study examined whether different proteins have different effects on whole-body protein turnover in adult rats. The rats were either starved, given a protein-free but energy-sufficient diet (1 MJ/kg body weight (BW) per d) or a diet containing intact casein, hydrolysed casein, or...... recovered from urinary ammonia and urea during isotope steady state for measurement of protein synthesis and protein degradation. Compared with starvation the protein-free diet decreased N excretion by 75%, probably by increasing the rate of reutilization of amino acids from endogenous proteins for protein...... synthesis. The protein diets produced a positive N balance which was independent of the protein source. Intact and hydrolysed casein increased protein synthesis 2.6- and 2.0-fold respectively, compared with the protein-free diet. Protein degradation increased 1.4- and 1.2-fold respectively. Hydrolysed soya...

  10. The evolutionary conserved oil body associated protein OBAP1 participates in the regulation of oil body size.

    Science.gov (United States)

    López-Ribera, Ignacio; La Paz, José Luis; Repiso, Carlos; García, Nora; Miquel, Mercè; Hernández, María Luisa; Martínez-Rivas, José Manuel; Vicient, Carlos M

    2014-03-01

    A transcriptomic approach has been used to identify genes predominantly expressed in maize (Zea mays) scutellum during maturation. One of the identified genes is oil body associated protein1 (obap1), which is transcribed during seed maturation predominantly in the scutellum, and its expression decreases rapidly after germination. Proteins similar to OBAP1 are present in all plants, including primitive plants and mosses, and in some fungi and bacteria. In plants, obap genes are divided in two subfamilies. Arabidopsis (Arabidopsis thaliana) genome contains five genes coding for OBAP proteins. Arabidopsis OBAP1a protein is accumulated during seed maturation and disappears after germination. Agroinfiltration of tobacco (Nicotiana benthamiana) epidermal leaf cells with fusions of OBAP1 to yellow fluorescent protein and immunogold labeling of embryo transmission electron microscopy sections showed that OBAP1 protein is mainly localized in the surface of the oil bodies. OBAP1 protein was detected in the oil body cellular fraction of Arabidopsis embryos. Deletion analyses demonstrate that the most hydrophilic part of the protein is responsible for the oil body localization, which suggests an indirect interaction of OBAP1 with other proteins in the oil body surface. An Arabidopsis mutant with a transfer DNA inserted in the second exon of the obap1a gene and an RNA interference line against the same gene showed a decrease in the germination rate, a decrease in seed oil content, and changes in fatty acid composition, and their embryos have few, big, and irregular oil bodies compared with the wild type. Taken together, our findings suggest that OBAP1 protein is involved in the stability of oil bodies. PMID:24406791

  11. Dietary protein content affects evolution for body size, body fat and viability in Drosophila melanogaster

    DEFF Research Database (Denmark)

    Kristensen, Torsten N; Overgaard, Johannes; Loeschcke, Volker;

    2011-01-01

    The ability to use different food sources is likely to be under strong selection if organisms are faced with natural variation in macro-nutrient (protein, carbohydrate and lipid) availabilities. Here, we use experimental evolution to study how variable dietary protein content affects adult body...... composition and developmental success in Drosophila melanogaster. We reared flies on either a standard diet or a protein-enriched diet for 17 generations before testing them on both diet types. Flies from lines selected on protein-rich diet produced phenotypes with higher total body mass and relative lipid...

  12. Protein components in saliva and plaque fluid from irradiated primates

    Energy Technology Data Exchange (ETDEWEB)

    Edgar, W.M.; Bowen, W.H.; Cole, M.F. (Caries Prevention and Research Branch, National Caries Program, NIDR, Bethesda, Maryland, USA)

    1982-01-01

    Irradiation of the major salivary glands of monkeys (Macaca mulatta) fed cariogenic diets leads to caries clinically indistinguishable from radiation caries in man. This study compares the organic compostion of individual samples of plaque fluid and saliva from irradiated and control monkeys receiving the same cariogenic diet. Plaque and saliva were collected from fasting, tranquillised animals. Four irradiated animals were sampled repeatedly as were non-irradiated controls. Total protein, albumin, immunoglobulins A, G, and M, and the third component of complement (C'3) were quantitated in plaque fluid and whole saliva. Salivary amylase and peroxidase activities were also determined. Plaque fluid and saliva samples were also subjected to polyacrylamide gel electrophoresis. The total viable anaerobic count and numbers of Streptococcus mutans were determined in samples of plaque. The results suggest that the major effect of irradiation leading to increased numbers of S. mutans and caries susceptibility is in the amount, and not the composition, of the saliva produced by the residual gland tissue. The scanty flow of saliva may reduce the effectiveness of cleansing, buffering and lubrication mechanisms as well as resulting in a marked reduction in the total amount of specific and non-specific immune factors entering the mouth.

  13. Protein components in saliva and plaque fluid from irradiated primates

    International Nuclear Information System (INIS)

    Irradiation of the major salivary glands of monkeys (Macaca mulatta) fed cariogenic diets leads to caries clinically indistinguishable from radiation caries in man. This study compares the organic compostion of individual samples of plaque fluid and saliva from irradiated and control monkeys receiving the same cariogenic diet. Plaque and saliva were collected from fasting, tranquillised animals. Four irradiated animals were sampled repeatedly as were non-irradiated controls. Total protein, albumin, immunoglobulins A, G, and M, and the third component of complement (C'3) were quantitated in plaque fluid and whole saliva. Salivary amylase and peroxidase activities were also determined. Plaque fluid and saliva samples were also subjected to polyacrylamide gel electrophoresis. The total viable anaerobic count and numbers of Streptococcus mutans were determined in samples of plaque. The results suggest that the major effect of irradiation leading to increased numbers of S. mutans and caries susceptibility is in the amount, and not the composition, of the saliva produced by the residual gland tissue. The scanty flow of saliva may reduce the effectiveness of cleansing, buffering and lubrication mechanisms as well as resulting in a marked reduction in the total amount of specific and non-specific immune factors entering the mouth. (author)

  14. Phosphorylation of an HP1-like protein is a prerequisite for heterochromatin body formation in Tetrahymena DNA elimination.

    Science.gov (United States)

    Kataoka, Kensuke; Noto, Tomoko; Mochizuki, Kazufumi

    2016-08-01

    Multiple heterochromatic loci are often clustered into a higher order nuclear architecture called a heterochromatin body in diverse eukaryotes. Although phosphorylation of Heterochromatin Protein 1 (HP1) family proteins regulates heterochromatin dynamics, its role in heterochromatin bodies remains unknown. We previously reported that dephosphorylation of the HP1-like protein Pdd1p is required for the formation of heterochromatin bodies during the process of programmed DNA elimination in the ciliated protozoan Tetrahymena Here, we show that the heterochromatin body component Jub4p is required for Pdd1p phosphorylation, heterochromatin body formation, and DNA elimination. Moreover, our analyses of unphosphorylatable Pdd1p mutants demonstrate that Pdd1p phosphorylation is required for heterochromatin body formation and DNA elimination, whereas it is dispensable for local heterochromatin assembly. Therefore, both phosphorylation and the following dephosphorylation of Pdd1p are necessary to facilitate the formation of heterochromatin bodies. We suggest that Jub4p-mediated phosphorylation of Pdd1p creates a chromatin environment that is a prerequisite for subsequent heterochromatin body assembly and DNA elimination. PMID:27466409

  15. Capillary gel electrophoresis for the quantification and purity determination of recombinant proteins in inclusion bodies.

    Science.gov (United States)

    Espinosa-de la Garza, Carlos E; Perdomo-Abúndez, Francisco C; Campos-García, Víctor R; Pérez, Néstor O; Flores-Ortiz, Luis F; Medina-Rivero, Emilio

    2013-09-01

    In this work, a high-resolution CGE method for quantification and purity determination of recombinant proteins was developed, involving a single-component inclusion bodies (IBs) solubilization solution. Different recombinant proteins expressed as IBs were used to show method capabilities, using recombinant interferon-β 1b as the model protein for method validation. Method linearity was verified in the range from 0.05 to 0.40 mg/mL and a determination coefficient (r(2) ) of 0.99 was obtained. The LOQs and LODs were 0.018 and 0.006 mg/mL, respectively. RSD for protein content repeatability test was 2.29%. In addition, RSD for protein purity repeatability test was 4.24%. Method accuracy was higher than 90%. Specificity was confirmed, as the method was able to separate recombinant interferon-β 1b monomer from other aggregates and impurities. Sample content and purity was demonstrated to be stable for up to 48 h. Overall, this method is suitable for the analysis of recombinant proteins in IBs according to the attributes established on the International Conference for Harmonization guidelines. PMID:23857606

  16. Features of somatotype and body weight component composition in patients with acne: boys and girls of Podillya region of Ukraine

    Directory of Open Access Journals (Sweden)

    Gunas Igor

    2016-06-01

    Full Text Available The article describes the differences in peculiarities of somatotype and body weight component composition in patients with acne, the study population being boys and girls of the Podillya region of Ukraine. In the study subjects, regardless of sex, body muscle mass, bone mass and the mesomorphic somatotype component of those with acne were significantly greater, while fat body mass indicators and the endomorphic somatotype component was smaller – in comparison to that of non-afflicted subjects of similar gender. Regarding the ectomorphic component of somatotype, between the surveyed groups of those with and without acne, whether male or female, no significant differences were revealed. For all indicators, whether the somatotype components or the component composition of body weight, between groups of boys or girls with different degrees of severity of acne, again no significant differences were established.

  17. Structure and Composition of Protein Bodies from Wild-Type and High-Lysine Barley Endosperm

    DEFF Research Database (Denmark)

    Ingversen, J.

    1975-01-01

    Protein bodies were isolated from 13 and 28 day old endosperms of barley mutant 1508 and its wild type, Bomi barley. The fine structure of the isolated protein bodies was determined by electron microscopy, and the proteins present in the preparations characterized by amino-acid analysis and SDS-p...

  18. BREN: Body Reflection Essence-Neuter Model for Separation of Reflection Components

    CERN Document Server

    Je, Changsoo

    2015-01-01

    We propose a novel reflection color model consisting of body essence and (mixed) neuter, and present an effective method for separating dichromatic reflection components using a single image. Body essence is an entity invariant to interface reflection, and has two degrees of freedom unlike hue and maximum chromaticity. As a result, the proposed method is insensitive to noise and proper for colors around CMY (cyan, magenta, and yellow) as well as RGB (red, green, and blue), contrary to the maximum chromaticity-based methods. Interface reflection is separated by using a Gaussian function, which removes a critical thresholding problem. Furthermore, the method does not require any region segmentation. Experimental results show the efficacy of the proposed model and method.

  19. Identification and characterization of protein interactions in the mammalian mRNA processing body using a novel two-hybrid assay

    International Nuclear Information System (INIS)

    Components of the mRNA processing body (P-body) regulate critical steps in mRNA storage, transport, translation and degradation. At the core of the P-body is the decapping complex, which removes the 5' cap from de-adenylated mRNAs and mediates an irreversible step in mRNA degradation. The assembly of P-bodies in Saccharomyces cerevisiae, Arabidopsis thaliana and Drosophila melanogaster has been previously described. Less is known about the assembly of mammalian P-bodies. To investigate the interactions that occur between components of mammalian P-bodies, we developed a fluorescence-based, two-hybrid assay system. The assay depends on the ability of one P-body component, fused to an exogenous nuclear localization sequence (NLS), to recruit other P-body components to the nucleus. The assay was used to investigate interactions between P-body components Ge-1, DCP2, DCP1, EDC3, RAP55, and RCK. The results of this study show that the modified two-hybrid assay can be used to identify protein interactions that occur in a macromolecular complex. The assay can also be used to efficiently detect protein interaction domains. The results provide important insights into mammalian P-body assembly and demonstrate similarities, and critical differences, between P-body assembly in mammalian cells compared with that of other species. -- Research highlights: → A two-hybrid assay was developed to study interactions in macromolecular complexes. → The assay was applied to interactions between components of mRNA P-bodies. → The assay effectively and efficiently identified protein interaction domains. → P-body assembly in mammalian cells differs from that in other species.

  20. Two Independent Mushroom Body Output Circuits Retrieve the Six Discrete Components of Drosophila Aversive Memory

    OpenAIRE

    Emna Bouzaiane; Séverine Trannoy; Lisa Scheunemann; Pierre-Yves Plaçais; Thomas Preat

    2015-01-01

    Understanding how the various memory components are encoded and how they interact to guide behavior requires knowledge of the underlying neural circuits. Currently, aversive olfactory memory in Drosophila is behaviorally subdivided into four discrete phases. Among these, short- and long-term memories rely, respectively, on the γ and α/β Kenyon cells (KCs), two distinct subsets of the ∼2,000 neurons in the mushroom body (MB). Whereas V2 efferent neurons retrieve memory from α/β KCs, the neuron...

  1. Disentangling the associations between parental BMI and offspring body composition using the four‐component model

    Science.gov (United States)

    Grijalva‐Eternod, Carlos; Cortina‐Borja, Mario; Williams, Jane; Fewtrell, Mary; Wells, Jonathan

    2016-01-01

    ABSTRACT Objectives This study sets out to investigate the intergenerational associations between the body mass index (BMI) of parents and the body composition of their offspring. Methods The cross‐sectional data were analyzed for 511 parent–offspring trios from London and south‐east England. The offspring were aged 5–21 years. Parental BMI was obtained by recall and offspring fat mass and lean mass were obtained using the four‐component model. Multivariable regression analysis, with multiple imputation for missing paternal values was used. Sensitivity analyses for levels of non‐paternity were conducted. Results A positive association was seen between parental BMI and offspring BMI, fat mass index (FMI), and lean mass index (LMI). The mother's BMI was positively associated with the BMI, FMI, and LMI z‐scores of both daughters and sons and of a similar magnitude for both sexes. The father's BMI showed similar associations to the mother's BMI, with his son's BMI, FMI, and LMI z‐scores, but no association with his daughter. Sensitivity tests for non‐paternity showed that maternal coefficients remained greater than paternal coefficients throughout but there was no statistical difference at greater levels of non‐paternity. Conclusions We found variable associations between parental BMI and offspring body composition. Associations were generally stronger for maternal than paternal BMI, and paternal associations appeared to differ between sons and daughters. In this cohort, the mother's BMI was statistically significantly associated with her child's body composition but the father's BMI was only associated with the body composition of his sons. Am. J. Hum. Biol. 28:524–533, 2016. © 2016 The Authors American Journal of Human Biology Published by Wiley Periodicals, Inc. PMID:26848813

  2. Mechanical component design for upgrading of whole body counter ND7500

    International Nuclear Information System (INIS)

    The Whole Body Counter (WBC) ND7500 is a bed type counting system that used for measuring radionuclide in the entire human body. Malaysian Nuclear Agency has this system, which savaged from Institute of Medical Research (IMR) in 1987. This system consists of a nuclear counting system and mechanical system that totally inoperable due to its counting system failures. In April 2003, both counting system and the mechanical system were tested. The mechanical component is working properly but needs some readjustment for the bed movement while for the counting system, only detectors can work but with a poor detecting capability. During IAEA expert visits on July 2003, both detectors were verified cannot be use any longer due to poor resolution and aging factor and a single (3 x 5 x 16) inches rectangular NaI(Tl) detector was then purchased in the end of 2004 to replace (3 x 5) inches cylindrical Na(Tl) detectors. The existing shielding cannot accommodate this new (3 x 5 x 16) inches dimension and the (5 x 16) inches detecting area. Therefore, shielding modification has been done based on effective detecting area and positioning test results. A new detector's entrance and detector stage were built at the bottom shielding. A new features, which is a detectors protection also been developed for detector safety. This upgrading task successfully accomplished as from experimental the design of positioning component can make system operated easily and also can give a good results to meets user's requirements. (Author)

  3. Identification and characterization of a novel Chlamydia trachomatis reticulate body protein

    DEFF Research Database (Denmark)

    Shaw, Allan C; Larsen, Martin Røssel; Roepstorff, Peter;

    2002-01-01

    identified by tandem mass spectrometry. No homology of this protein was observed to proteins from other organisms. The protein was conserved in C. trachomatis but not found in Chlamydia pneumoniae. Using proteomics, we show that the expression of the protein is initiated at the middle of the developmental...... cycle. The protein is rapidly degraded and is only present in reticulate or intermediate bodies, suggesting a possible function in the intracellular stage of C. trachomatis development. We have termed the protein '7-kDa reticulate body protein'....

  4. Interaction preferences across protein-protein interfaces of obligatory and non-obligatory components are different

    Directory of Open Access Journals (Sweden)

    Srinivasan N

    2005-08-01

    Full Text Available Abstract Background A polypeptide chain of a protein-protein complex is said to be obligatory if it is bound to another chain throughout its functional lifetime. Such a chain might not adopt the native fold in the unbound form. A non-obligatory polypeptide chain associates with another chain and dissociates upon molecular stimulus. Although conformational changes at the interaction interface are expected, the overall 3-D structure of the non-obligatory chain is unaltered. The present study focuses on protein-protein complexes to understand further the differences between obligatory and non-obligatory interfaces. Results A non-obligatory chain in a complex of known 3-D structure is recognized by its stable existence with same fold in the bound and unbound forms. On the contrary, an obligatory chain is detected by its existence only in the bound form with no evidence for the native-like fold of the chain in the unbound form. Various interfacial properties of a large number of complexes of known 3-D structures thus classified are comparatively analyzed with an aim to identify structural descriptors that distinguish these two types of interfaces. We report that the interaction patterns across the interfaces of obligatory and non-obligatory components are different and contacts made by obligatory chains are predominantly non-polar. The obligatory chains have a higher number of contacts per interface (20 ± 14 contacts per interface than non-obligatory chains (13 ± 6 contacts per interface. The involvement of main chain atoms is higher in the case of obligatory chains (16.9 % compared to non-obligatory chains (11.2 %. The β-sheet formation across the subunits is observed only among obligatory protein chains in the dataset. Apart from these, other features like residue preferences and interface area produce marginal differences and they may be considered collectively while distinguishing the two types of interfaces. Conclusion These results can be

  5. Nigella sativa oil for the control of irradiation induced disorders in serum protein components of male albino rats

    International Nuclear Information System (INIS)

    The effect of whole body gamma irradiation at the dose level 7 Gy delivered as shot dose or cumulative doses, on the electrophoretic pattern of serum protein components, was studied in male albino rats subjected to Nigella sativa treatment. Sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE) of serum proteins of irradiated rats showed significant increases in percentages of some fractions of high molecular weights. Administration of nigella sativa oil before radiation exposure resulted in detectable radioprotective effect. Radiation protection effect exerted by nigella sativa oil prior to cumulative doses was more pronounced than that in case of one shot dose

  6. Prolonged bed rest decreases skeletal muscle and whole body protein synthesis

    Science.gov (United States)

    Ferrando, A. A.; Lane, H. W.; Stuart, C. A.; Davis-Street, J.; Wolfe, R. R.

    1996-01-01

    We sought to determine the extent to which the loss of lean body mass and nitrogen during inactivity was due to alterations in skeletal muscle protein metabolism. Six male subjects were studied during 7 days of diet stabilization and after 14 days of stimulated microgravity (-6 degrees bed rest). Nitrogen balance became more negative (P muscle protein synthesis (PS; P muscle protein also decreased by 46% (P protein breakdown and inward transport. Whole body protein synthesis determined by [15N]alanine ingestion on six subjects also revealed a 14% decrease (P protein breakdown change significantly. These results indicate that the loss of body protein with inactivity is predominantly due to a decrease in muscle PS and that this decrease is reflected in both whole body and skeletal muscle measures.

  7. Purification and protein composition of oil bodies from Brassica napus seeds

    Directory of Open Access Journals (Sweden)

    Jolivet Pascale

    2006-11-01

    Full Text Available Seed oil bodies are intracellular particles to store lipids as food reserves in oleaginous plants. Description of oil body-associated proteins of Arabidopsis thaliana has been recently reported whereas only few data are available in the case of rapeseed. Oil bodies have been prepared from two double-low varieties of Brassica napus seeds, a standard variety (Explus and an oleic variety (Cabriolet. Oil bodies have been purified using floatation technique in the successive presence of high salt concentration, detergent or urea in order to remove non-specifically trapped proteins. The integrity of the oil bodies has been verified and their size estimated. Their protein and fatty acid contents have been determined. The proteins composing these organelles were extracted, separated by denaturing gel electrophoresis, digested by trypsin and their peptides were subsequently analyzed by liquid chromatography-tandem mass spectrometry. Protein identification was performed using Arabidopsis thaliana protein sequence database and a collection of Expressed Sequence Tag (EST of Brassica napus generated from the framework of the French plant genomics programme “Genoplante”. This led to the identification of a limited number of proteins: eight oleosins showing a high similarity each other and representing up to 75% of oil body proteins, a 11 β hydroxysteroid dehydrogenase-like protein highly homologous to the same protein from A. thaliana, and only few contaminating proteins associated with myrosinase activity.

  8. Numerical code for multi-component galaxies: from N-body to chemistry and magnetic fields

    CERN Document Server

    Khoperskov, S A; Khoperskov, A V; Lubimov, V N

    2015-01-01

    We present a numerical code for multi-component simulation of the galactic evolution. Our code includes the following parts: $N$-body is used to evolve dark matter, stellar dynamics and dust grains, gas dynamics is based on TVD-MUSCL scheme with the extra modules for thermal processes, star formation, magnetic fields, chemical kinetics and multi-species advection. We describe our code in brief, but we give more details for the magneto-gas dynamics. We present several tests for our code and show that our code have passed the tests with a reasonable accuracy. Our code is parallelized using the MPI library. We apply our code to study the large scale dynamics of galactic discs.

  9. Avoidance of crack inducement when laser welding hot-formed car body components - a variable analysis

    Science.gov (United States)

    Larsson, Johnny K.

    The Volvo XC60 car body contains numerous parts in Ultra High Strength Steels (UHSS) in order to guarantee the structural integrity of the car in the event of a crash situation. Most of the parts are manufactured in a hot-forming process, so called presshardening, resulting in component tensile strength in the range of 1,500 MPa. As this type of material also presents fairly high carbon content (˜0.22%) it brings a challenge when it comes to welding. The Volvo XC60 car body is at the same time to a large extent assembled by laser welding technology. In early development stages of the project (Y413), it was observed that laser welding of hot-formed components presented a number of challenges due to the unique conditions offered by this welding method. The presentation will thoroughly describe the modes of procedure how to avoid crack inducement during the welding operation. A variable analysis approach was used based on the present circumstances at the production facility in the Gent plant. Crucial variables at laser welding such as gap between sheets, focal point position, welding speed and laser weld position relative to the flange edge were included in a test matrix and welding trials were carried out accordingly in the Pilot Plant in Gothenburg. The paper will discuss those welding results, the subsequent analysis and plausible theoretic explanations. From the lessons learnt in this research, the optimum laser welding parameters were then transferred to the laser welding stations in the Gent plant. There it has been proven, that also at high volume automotive manufacturing, it is possible to provide an outstanding weld quality also at such difficult pre-conditions. The presentation ends with some facts and figures and experiences from high volume series production, which also includes aspects on quality assurance.

  10. Oil bodies and their associated proteins, oleosin and caleosin

    DEFF Research Database (Denmark)

    Frandsen, Gitte I.; Mundy, John; Tzen, Jason T. C.

    2001-01-01

    bodies have been described, oleosin and caleosin. Oleosin is thought to be important for oil body stabilization in the cytosol, although neither the structure nor the function of oleosin has been fully elucidated. Even less is known about caleosin, which has only recently been described [Chen et al...... relate to structural and functional aspects of oleosin and caleosin....

  11. Two independent mushroom body output circuits retrieve the six discrete components of Drosophila aversive memory.

    Science.gov (United States)

    Bouzaiane, Emna; Trannoy, Séverine; Scheunemann, Lisa; Plaçais, Pierre-Yves; Preat, Thomas

    2015-05-26

    Understanding how the various memory components are encoded and how they interact to guide behavior requires knowledge of the underlying neural circuits. Currently, aversive olfactory memory in Drosophila is behaviorally subdivided into four discrete phases. Among these, short- and long-term memories rely, respectively, on the γ and α/β Kenyon cells (KCs), two distinct subsets of the ∼2,000 neurons in the mushroom body (MB). Whereas V2 efferent neurons retrieve memory from α/β KCs, the neurons that retrieve short-term memory are unknown. We identified a specific pair of MB efferent neurons, named M6, that retrieve memory from γ KCs. Moreover, our network analysis revealed that six discrete memory phases actually exist, three of which have been conflated in the past. At each time point, two distinct memory components separately recruit either V2 or M6 output pathways. Memory retrieval thus features a dramatic convergence from KCs to MB efferent neurons. PMID:25981036

  12. Two Independent Mushroom Body Output Circuits Retrieve the Six Discrete Components of Drosophila Aversive Memory

    Directory of Open Access Journals (Sweden)

    Emna Bouzaiane

    2015-05-01

    Full Text Available Understanding how the various memory components are encoded and how they interact to guide behavior requires knowledge of the underlying neural circuits. Currently, aversive olfactory memory in Drosophila is behaviorally subdivided into four discrete phases. Among these, short- and long-term memories rely, respectively, on the γ and α/β Kenyon cells (KCs, two distinct subsets of the ∼2,000 neurons in the mushroom body (MB. Whereas V2 efferent neurons retrieve memory from α/β KCs, the neurons that retrieve short-term memory are unknown. We identified a specific pair of MB efferent neurons, named M6, that retrieve memory from γ KCs. Moreover, our network analysis revealed that six discrete memory phases actually exist, three of which have been conflated in the past. At each time point, two distinct memory components separately recruit either V2 or M6 output pathways. Memory retrieval thus features a dramatic convergence from KCs to MB efferent neurons.

  13. Body composition and deposition efficiency of protein and energy in grazing young bulls

    OpenAIRE

    Eriton Egidio Lisboa Valente; Mário Fonseca Paulino; Marcos Inácio Marcondes; Isabela Fernanda Teixeira Dias

    2014-01-01

    The effects of supplementation with different protein: carbohydrate ratios on body composition, carcass characteristics and protein and energy deposition efficiency of young were assessed. Twenty-four Nellorecalves (132.5 ± 5.5 kgand 90-150 days of age) were kept on pasture for a 430 day experimental period. The treatments were: Control = mineral mixture only; HPHC = high-protein and high-carbohydrate supplement; HPLC = high-protein and low-carbohydrate supplement; LPHC = low-protein and high...

  14. Solvent content of protein crystals from diffraction intensities by Independent Component Analysis

    OpenAIRE

    Lamura, Antonio; Ladisa, Massimo; Nico, Giovanni; Siliqi, Dritan

    2003-01-01

    An analysis of the protein content of several crystal forms of proteins has been performed. We apply a new numerical technique, the Independent Component Analysis (ICA), to determine the volume fraction of the asymmetric unit occupied by the protein. This technique requires only the crystallographic data of structure factors as input.

  15. The study of dual energy X-ray absorptiometry on body composition components in obesity

    International Nuclear Information System (INIS)

    Objective: To study the correlation of the bone mineral density (BMD) and the body composition components of body mass index (BMI), FAT and LEAN in Chinese obesity. Methods: There were 150 cases in obesity group diagnosed by BMI, including 75 males [ median age 46 years, mean weight (89.64 ± 8.33) kg] and 75 females [median age 45 years, mean weight (77.23 ± 6.85) kg]. There were 150 persons with normal BMI in the control group, including 75 males [(median age 47 years, mean weight (62.34 ± 5.72) kg] and 75 females [ median age 45 years, mean weight (50.16 ± 5.06) kg]. The body height and weight of 300 persons in two groups were measured respectively and ,simultaneously calculated the BMI. These data and the body composition parameters measured by the dual energy X-ray absorptiometry (DEXA) in these two groups were compared and analyzed. The data obtained used two-sample t-test analysis, bi-variable correlation used Pearson linear correlation analysis and multi-variable correlation used multiple linear regression analysis. Results: FAT of arms, legs trunk and total body of male cases in obesity group was (2.90 ± 0.57), (7.48 ± 1.46), (15.67 ± 3.05), (30.92 ± 5.94) kg respectively, FAT% was (30.9 ± 5.1)%, (30.6 ± 5.8)%, (37.3 ± 4.7)%, (35.1 ± 4.4)% respectively, it was significantly higher than that in control group [FAT was (1.12 ± 0.64), (3.27 ± 1.22), (6.71 ± 3.29), (11.61 ± 5.16) kg respectively, FAT% was (15.4 ± 4.8)%, (16.5 ± 5.0)%, (21.8 ± 5.8)%, (18.6 ± 5.3)% respectively], P 2 respectively, it was significantly higher than that in control group [ LEAN was (22.89 ± 1.68), (48.89 ± 3.72) kg respectively, BMD was (0.89 ± 0.07), (1.15 ± 0.06) g/cm2 respectively], P 2 respectively, and there were no statistical significance compared with those in control group [LEAN was (5.99 ± 0.72), (16.83 ± 1.67) kg respectively, BMD was (0.90 ± 0.08), (1.29 ± 0.09) g/cm2 respectively]. FAT of arms, legs, trunk and total body of females in obesity

  16. Daytime pattern of post-exercise protein intake affects whole-body protein turnover in resistance-trained males

    OpenAIRE

    Moore Daniel R; Areta Jose; Coffey Vernon G; Stellingwerff Trent; Phillips Stuart M; Burke Louise M; Cléroux Marilyn; Godin Jean-Philippe; Hawley John A

    2012-01-01

    Abstract Background The pattern of protein intake following exercise may impact whole-body protein turnover and net protein retention. We determined the effects of different protein feeding strategies on protein metabolism in resistance-trained young men. Methods Participants were randomly assigned to ingest either 80g of whey protein as 8x10g every 1.5h (PULSE; n=8), 4x20g every 3h (intermediate, INT; n=7), or 2x40g every 6h (BOLUS; n=8) after an acute bout of bilateral knee extension exerci...

  17. Ret Finger Protein: An E3 Ubiquitin Ligase Juxtaposed to the XY Body in Meiosis

    Directory of Open Access Journals (Sweden)

    Isabelle Gillot

    2009-01-01

    Full Text Available During prophase I of male meiosis, the sex chromosomes form a compact structure called XY body that associates with the nuclear membrane of pachytene spermatocytes. Ret Finger Protein is a transcriptional repressor, able to interact with both nuclear matrix-associated proteins and double-stranded DNA. We report the precise and unique localization of Ret Finger Protein in pachytene spermatocytes, in which Ret Finger Protein takes place of lamin B1, between the XY body and the inner nuclear membrane. This localization of Ret Finger Protein does not seem to be associated with O-glycosylation or sumoylation. In addition, we demonstrate that Ret Finger Protein contains an E3 ubiquitin ligase activity. These observations lead to an attractive hypothesis in which Ret Finger Protein would be involved in the positioning and the attachment of XY body to the nuclear lamina of pachytene spermatocytes.

  18. Mba1, a Novel Component of the Mitochondrial Protein Export Machinery of the Yeast Saccharomyces cerevisiae

    OpenAIRE

    Preuss, Marc; Leonhard, Klaus; Hell, Kai; Stuart, Rosemary A.; Neupert, Walter; Herrmann, Johannes M.

    2001-01-01

    The biogenesis of mitochondria requires the integration of many proteins into the inner membrane from the matrix side. The inner membrane protein Oxa1 plays an important role in this process. We identified Mba1 as a second mitochondrial component that is required for efficient protein insertion. Like Oxa1, Mba1 specifically interacts both with mitochondrial translation products and with conservatively sorted, nuclear-encoded proteins during their integration into the inner membrane. Oxa1 and ...

  19. Casein and soy protein meals differentially affect whole-body and splanchnic protein metabolism in healthy humans.

    Science.gov (United States)

    Luiking, Yvette C; Deutz, Nicolaas E P; Jäkel, Martin; Soeters, Peter B

    2005-05-01

    Dietary protein quality is considered to be dependent on the degree and velocity with which protein is digested, absorbed as amino acids, and retained in the gut as newly synthesized protein. Metabolic animal studies suggest that the quality of soy protein is inferior to that of casein protein, but confirmatory studies in humans are lacking. The study objective was to assess the quality of casein and soy protein by comparing their metabolic effects in healthy human subjects. Whole-body protein kinetics, splanchnic leucine extraction, and urea production rates were measured in the postabsorptive state and during 8-h enteral intakes of isonitrogenous [0.42 g protein/(kg body weight . 8 h)] protein-based test meals, which contained either casein (CAPM; n = 12) or soy protein (SOPM; n = 10) in 2 separate groups. Stable isotope techniques were used to study metabolic effects. With enteral food intake, protein metabolism changed from net protein breakdown to net protein synthesis. Net protein synthesis was greater in the CAPM group than in the SOPM group [52 +/- 14 and 17 +/- 14 nmol/(kg fat-free mass (FFM) . min), respectively; P CAPM (P = 0.07). Absolute splanchnic extraction of leucine was higher in the subjects that consumed CAPM [306 +/- 31 nmol/(kg FFM . min)] vs. those that consumed SOPM [235 +/- 29 nmol/(kg FFM . min); P < 0.01]. In conclusion, a significantly larger portion of soy protein is degraded to urea, whereas casein protein likely contributes to splanchnic utilization (probably protein synthesis) to a greater extent. The biological value of soy protein must be considered inferior to that of casein protein in humans. PMID:15867285

  20. Daytime pattern of post-exercise protein intake affects whole-body protein turnover in resistance-trained males

    Directory of Open Access Journals (Sweden)

    Moore Daniel R

    2012-10-01

    Full Text Available Abstract Background The pattern of protein intake following exercise may impact whole-body protein turnover and net protein retention. We determined the effects of different protein feeding strategies on protein metabolism in resistance-trained young men. Methods Participants were randomly assigned to ingest either 80g of whey protein as 8x10g every 1.5h (PULSE; n=8, 4x20g every 3h (intermediate, INT; n=7, or 2x40g every 6h (BOLUS; n=8 after an acute bout of bilateral knee extension exercise (4x10 repetitions at 80% maximal strength. Whole-body protein turnover (Q, synthesis (S, breakdown (B, and net balance (NB were measured throughout 12h of recovery by a bolus ingestion of [15N]glycine with urinary [15N]ammonia enrichment as the collected end-product. Results PULSE Q rates were greater than BOLUS (~19%, P Conclusion We conclude that the pattern of ingested protein, and not only the total daily amount, can impact whole-body protein metabolism. Individuals aiming to maximize NB would likely benefit from repeated ingestion of moderate amounts of protein (~20g at regular intervals (~3h throughout the day.

  1. Essential roles of protein-solvent many-body correlation in solvent-entropy effect on protein folding and denaturation: Comparison between hard-sphere solvent and water

    Science.gov (United States)

    Oshima, Hiraku; Kinoshita, Masahiro

    2015-04-01

    In earlier works, we showed that the entropic effect originating from the translational displacement of water molecules plays the pivotal role in protein folding and denaturation. The two different solvent models, hard-sphere solvent and model water, were employed in theoretical methods wherein the entropic effect was treated as an essential factor. However, there were similarities and differences in the results obtained from the two solvent models. In the present work, to unveil the physical origins of the similarities and differences, we simultaneously consider structural transition, cold denaturation, and pressure denaturation for the same protein by employing the two solvent models and considering three different thermodynamic states for each solvent model. The solvent-entropy change upon protein folding/unfolding is decomposed into the protein-solvent pair (PA) and many-body (MB) correlation components using the integral equation theories. Each component is further decomposed into the excluded-volume (EV) and solvent-accessible surface (SAS) terms by applying the morphometric approach. The four physically insightful constituents, (PA, EV), (PA, SAS), (MB, EV), and (MB, SAS), are thus obtained. Moreover, (MB, SAS) is discussed by dividing it into two factors. This all-inclusive investigation leads to the following results: (1) the protein-water many-body correlation always plays critical roles in a variety of folding/unfolding processes; (2) the hard-sphere solvent model fails when it does not correctly reproduce the protein-water many-body correlation; (3) the hard-sphere solvent model becomes problematic when the dependence of the many-body correlation on the solvent number density and temperature is essential: it is not quite suited to studies on cold and pressure denaturating of a protein; (4) when the temperature and solvent number density are limited to the ambient values, the hard-sphere solvent model is usually successful; and (5) even at the ambient

  2. Essential roles of protein-solvent many-body correlation in solvent-entropy effect on protein folding and denaturation: Comparison between hard-sphere solvent and water

    Energy Technology Data Exchange (ETDEWEB)

    Oshima, Hiraku; Kinoshita, Masahiro, E-mail: kinoshit@iae.kyoto-u.ac.jp [Institute of Advanced Energy, Kyoto University, Uji, Kyoto 611-0011 (Japan)

    2015-04-14

    In earlier works, we showed that the entropic effect originating from the translational displacement of water molecules plays the pivotal role in protein folding and denaturation. The two different solvent models, hard-sphere solvent and model water, were employed in theoretical methods wherein the entropic effect was treated as an essential factor. However, there were similarities and differences in the results obtained from the two solvent models. In the present work, to unveil the physical origins of the similarities and differences, we simultaneously consider structural transition, cold denaturation, and pressure denaturation for the same protein by employing the two solvent models and considering three different thermodynamic states for each solvent model. The solvent-entropy change upon protein folding/unfolding is decomposed into the protein-solvent pair (PA) and many-body (MB) correlation components using the integral equation theories. Each component is further decomposed into the excluded-volume (EV) and solvent-accessible surface (SAS) terms by applying the morphometric approach. The four physically insightful constituents, (PA, EV), (PA, SAS), (MB, EV), and (MB, SAS), are thus obtained. Moreover, (MB, SAS) is discussed by dividing it into two factors. This all-inclusive investigation leads to the following results: (1) the protein-water many-body correlation always plays critical roles in a variety of folding/unfolding processes; (2) the hard-sphere solvent model fails when it does not correctly reproduce the protein-water many-body correlation; (3) the hard-sphere solvent model becomes problematic when the dependence of the many-body correlation on the solvent number density and temperature is essential: it is not quite suited to studies on cold and pressure denaturating of a protein; (4) when the temperature and solvent number density are limited to the ambient values, the hard-sphere solvent model is usually successful; and (5) even at the ambient

  3. Essential roles of protein-solvent many-body correlation in solvent-entropy effect on protein folding and denaturation: Comparison between hard-sphere solvent and water

    International Nuclear Information System (INIS)

    In earlier works, we showed that the entropic effect originating from the translational displacement of water molecules plays the pivotal role in protein folding and denaturation. The two different solvent models, hard-sphere solvent and model water, were employed in theoretical methods wherein the entropic effect was treated as an essential factor. However, there were similarities and differences in the results obtained from the two solvent models. In the present work, to unveil the physical origins of the similarities and differences, we simultaneously consider structural transition, cold denaturation, and pressure denaturation for the same protein by employing the two solvent models and considering three different thermodynamic states for each solvent model. The solvent-entropy change upon protein folding/unfolding is decomposed into the protein-solvent pair (PA) and many-body (MB) correlation components using the integral equation theories. Each component is further decomposed into the excluded-volume (EV) and solvent-accessible surface (SAS) terms by applying the morphometric approach. The four physically insightful constituents, (PA, EV), (PA, SAS), (MB, EV), and (MB, SAS), are thus obtained. Moreover, (MB, SAS) is discussed by dividing it into two factors. This all-inclusive investigation leads to the following results: (1) the protein-water many-body correlation always plays critical roles in a variety of folding/unfolding processes; (2) the hard-sphere solvent model fails when it does not correctly reproduce the protein-water many-body correlation; (3) the hard-sphere solvent model becomes problematic when the dependence of the many-body correlation on the solvent number density and temperature is essential: it is not quite suited to studies on cold and pressure denaturating of a protein; (4) when the temperature and solvent number density are limited to the ambient values, the hard-sphere solvent model is usually successful; and (5) even at the ambient

  4. Supplementation with a fish protein hydrolysate (Micromesistius poutassou): effects on body weight, body composition, and CCK/GLP-1 secretion

    Science.gov (United States)

    Nobile, Vincenzo; Duclos, Elisa; Michelotti, Angela; Bizzaro, Gioia; Negro, Massimo; Soisson, Florian

    2016-01-01

    Background Fish protein hydrolysates (FPHs) have been reported as a suitable source of proteins for human nutrition because of their balanced amino acid composition and positive effect on gastrointestinal absorption. Objective Here, we investigated the effect of a FPH, Slimpro®, obtained from blue whiting (Micromesistius poutassou) muscle by enzymatic hydrolysis, on body composition and on stimulating cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1) secretion. Design A randomized clinical study was carried out on 120, slightly overweight (25 kg/m2 ≤ BMIProduct use was associated with a mild hypocaloric diet (−300 kcal/day). Body composition (body weight; fat mass; extracellular water; and circumference of waist, thighs, and hips) and CCK/GLP-1 blood levels were measured at the beginning of the study and after 45 and 90 days of product use. CCK/GLP-1 levels were measured since they are involved in controlling food intake. Results Treated subjects reported an improvement of body weight composition and an increased blood concentration of both CCK and GLP-1. No differences were found between the 1.4 and 2.8 g FPH doses, indicating a plateau effect starting from 1.4 g FPH. Conclusions Both 1.4 and 2.8 g of FPH were effective in improving body composition and in increasing CCK and GLP-1 blood levels. PMID:26829186

  5. Dynamics of the component of body composition in athletes playing sports with damage to the medial meniscus of the knee

    OpenAIRE

    Moh'd Khalil Moh'd Abdel Kader.

    2012-01-01

    The question of the influence of the developed program of physical rehabilitation with the use of modern means and methods of recovery on the performance component of body composition in athletes playing sports after arthroscopic stapling the medial meniscus was considered. The analysis and synthesis of scientific and methodological data on the determination of body composition analysis techniques using bioelectrical resistance was shown. Found that long-term immobilization that accompanies t...

  6. Relationship of Heath and Carter's Second Component to Lean Body Mass and Height in College Women

    Science.gov (United States)

    Slaughter, M. H.; And Others

    1977-01-01

    The Heath and Carter approach to determining somatotypes is less accurate than is regression analysis, mainly because of the lack of association between skeletal widths and lean body mass as measured by body density and whole-body fat percentage, holding constant muscle circumference. (Author)

  7. Comparative study of contents of several bioactive components in fruiting bodies and mycelia of culinary-medicinal mushrooms.

    Science.gov (United States)

    Lo, Yu-Chang; Lin, Shin-Yi; Ulziijargal, Enkhjargal; Chen, Shin-Yu; Chien, Rao-Chi; Tzou, Yi-Jing; Mau, Jeng-Leun

    2012-01-01

    Mushrooms have been consumed for thousands of years, and several bioactive components were found therein, including lovastatin, γ-aminobutyric acid (GABA) and ergothioneine. The study reported herein was to analyze these three bioactive components in 15 fruiting bodies and 9 mycelia of 19 species of mushrooms from genera Agaricus, Agrocybe, Auricularia, Boletus, Ganoderma, Hypsizygus, Inonotus, Lentinus, Morchella, Pleurotus, Tremella, Termitomyces, and Volvariella. The results show that Hypsizygus marmoreus contained the highest amount of lovastatin (628.05 mg/kg) in fruiting bodies and Morchella esculenta contained the highest amount (1438.42 mg/ kg) in mycelia. Agaricus brasiliensis contained the highest amount of GABA (1844.85 mg/kg) in fruiting bodies, and mycelia of Boletus edulis, Pleurotus citrinopileatus, and Termitomyces albuminosus contained extraordinarily higher amounts (1274.03, 1631.67, and 2560.00 mg/kg, respectively). Volvariella volvacea contained the highest amount of ergothioneine (537.27 mg/kg) in fruiting bodies and mycelia; Boletus edulis, Pleurotus ferulae, and P. salmoneostramineus contained relatively higher amount of ergothioneine too (258.03, 250.23, and 222.08 mg/kg, respectively). However, none of these components was detected in fruiting bodies of Inonotus obliquus. In conclusion, these three bioactive components were commonly found in most mushrooms, and these results might be related to their beneficial effects. PMID:23510173

  8. Crystallographic characterization of a multidomain histidine protein kinase from an essential two-component regulatory system

    OpenAIRE

    Zhao, Haiyan; Tang, Liang

    2009-01-01

    The multidomain cytoplasmic portion of the histidine protein kinase from an essential two-component signal transduction system has been crystallized and X-ray data have been collected to 2.8 Å resolution.

  9. The effect of protein and glycemic index on children's body composition

    DEFF Research Database (Denmark)

    Papadaki, Angeliki; Linardakis, Manolis; Larsen, Thomas Meinert;

    2010-01-01

    To investigate the effect of protein and glycemic index (GI) on body composition among European children in the randomized, 6-month dietary intervention DiOGenes (diet, obesity, and genes) family-based study.......To investigate the effect of protein and glycemic index (GI) on body composition among European children in the randomized, 6-month dietary intervention DiOGenes (diet, obesity, and genes) family-based study....

  10. Sensitivity and Specificity of Body Mass Index as a Definition of the Obesity Component of Metabolic Syndrome

    OpenAIRE

    M. Chakraborty, Bandana; Chakraborty, Ranajit

    2007-01-01

    Metabolic syndrome (MS) is a combination of risk factors that are associated with several chronic diseases. Its components (obesity, dyslipidemia, carbohydrate intolerance, hypertension, microalbumineria) are diverse, whose thresholds vary in different definitions of MS. For example, a World Health Organization (WHO) panel defined the obesity component of MS based on waist-hip ratio, or body mass index (BMI), while the National Cholesterol Education Program (NCEP) defined the obes...

  11. Low Dietary Protein Status Potentiating Risk of Health Hazard in Whole Body Gamma Irradiated Rats

    International Nuclear Information System (INIS)

    Investigations were planned to assess the changes in certain biochemical parameters as affected by the synergistic effect of exposure to fractionated doses of rays and / or feeding on different protein levels. The date showed that animals kept on normal or low protein diet exhibited a significant decrease in serum total protein and glucose. Also , a significant increase was recorded in insulin level in rats exposed at the radiation dose level of 20 Gy. Exposure to cumulative doses of irradiation has aggrevated the hyperglycemic effect of high protein diet with a significant and marked increase of insulin at all the applied doses. Animals fed normal high or low protein diet were found to exert significant decreases in T3, T4 while a significant increase in TSH of high protein group occurred as a result of exposure to cumulative doses of gamma-irradiation. Rats kept on low protein diet exhibited losses in body weight, hypercholesterolemia, low levels of phospholipids and triglycerides as compared with the normal protein diet group. In contrast high protein diet group showed no serious effects. Irradiation has potentiated body weight losses, hypotriglyceridemia and hypercholesterolemia in animal group fed low protein diet with a significant increase in serum phospholipids due to the higher radiation dose of 20 Gy. Protein deficiency acted synergistically with gamma irradiation and increased the susceptibility of body organs to radiation damage. Such findings contributed to the knowledge which stimulated the decrease of the internationally recognized occupational dose limits from 50 down to 20 m Sv (ICRP 1991)

  12. The efects of whey protein and resistant starch on body weight

    Directory of Open Access Journals (Sweden)

    Fatemah Malekian

    2015-08-01

    Full Text Available Background: Obesity is widely recognized as one of the most critical health threats to families and children across the country. Obesity is a very serious health problem for people in Louisiana and especially in African Americans Children with 40.5% in the State classified as either overweight or obese as compared to 41.2% nationally. African American women have the highest rates of being overweight or obese (79.8% compared to non-Hispanic whites (57.9%. In 2007, African Americans were 1.4 times as likely to be obese as whites. Therefore this study was designed to test the impact of dietary whey protein (WP and resistant starch (RS shakes/smoothies on reduction of body fat via increased satiety and increased energy expenditure in African Americans. Methods: Twenty eight African American males and females, age 21-43 years old were randomly divided into two groups. For a period of 24-weeks, the treatment group consumed WP and RS shakes/smoothies for breakfast and received nutrition education. For the same period, the control group consumed the same shake/smoothies but with starch powder and received nutrition education. The data was analyzed using SAS version 9.3. Results: At the end of the 24 week study, the treatment group lost a mean body weight of approximately 7 kg (P ≤ 0.008 that was significantly different from the control group (P ≤ 0.209. In addition, the treatment group exhibited a significant decrease of ~6 cm in waist circumference (P ≤ 0.023. There was no significant effect on mean blood pressure in treatment and control group. Conclusion: The findings from this study suggest that a combination of WP and RS in the form of shake/smoothie consumed for breakfast along with a nutrition education component may be an effective method in decreasing body weight, improving waist circumferences and cumulative food intake in African American males and females.

  13. RNA-binding proteins in eye development and disease: implication of conserved RNA granule components.

    Science.gov (United States)

    Dash, Soma; Siddam, Archana D; Barnum, Carrie E; Janga, Sarath Chandra; Lachke, Salil A

    2016-07-01

    The molecular biology of metazoan eye development is an area of intense investigation. These efforts have led to the surprising recognition that although insect and vertebrate eyes have dramatically different structures, the orthologs or family members of several conserved transcription and signaling regulators such as Pax6, Six3, Prox1, and Bmp4 are commonly required for their development. In contrast, our understanding of posttranscriptional regulation in eye development and disease, particularly regarding the function of RNA-binding proteins (RBPs), is limited. We examine the present knowledge of RBPs in eye development in the insect model Drosophila as well as several vertebrate models such as fish, frog, chicken, and mouse. Interestingly, of the 42 RBPs that have been investigated for their expression or function in vertebrate eye development, 24 (~60%) are recognized in eukaryotic cells as components of RNA granules such as processing bodies, stress granules, or other specialized ribonucleoprotein (RNP) complexes. We discuss the distinct developmental and cellular events that may necessitate potential RBP/RNA granule-associated RNA regulon models to facilitate posttranscriptional control of gene expression in eye morphogenesis. In support of these hypotheses, three RBPs and RNP/RNA granule components Tdrd7, Caprin2, and Stau2 are linked to ocular developmental defects such as congenital cataract, Peters anomaly, and microphthalmia in human patients or animal models. We conclude by discussing the utility of interdisciplinary approaches such as the bioinformatics tool iSyTE (integrated Systems Tool for Eye gene discovery) to prioritize RBPs for deriving posttranscriptional regulatory networks in eye development and disease. WIREs RNA 2016, 7:527-557. doi: 10.1002/wrna.1355 For further resources related to this article, please visit the WIREs website. PMID:27133484

  14. Measurement of body fluid proteins by polyacrylamide gel electrophoresis

    OpenAIRE

    Johnson, MH; Thompson, EJ

    1982-01-01

    A development of the disc polyacrylamide gel electrophoretic system of Ornstein and Davis, which has been applied to the analysis of unconcentrated cerebrospinal fluid, is described. Modifications to the scanning densitometer have improved the signal:noise ratio of the gel scanning system. Using this technique, we have studied the dye-binding properties of albumin, and of beta- and gamma-globulins, and have shown that reproducible quantification of proteins can be achieved. The advantages of ...

  15. Insulin and amino acids stimulate whole body protein synthesis in neonates

    Science.gov (United States)

    Insulin and amino acids (AA) stimulate muscle protein synthesis in neonatal pigs. To determine the effects of insulin and AA on whole body protein turnover, hyperinsulinemic (0 and 100 ng/(kg[0.66]/min))-euglycemic-AA clamps were performed during euaminoacidemia or hyperaminoacidemia in fasted 7-d-...

  16. Effects of Dietary Male and Female Ferula Eleaochytris Powder on Growth Performance and Body Components of Broiler Chicks

    Directory of Open Access Journals (Sweden)

    Metin Duru

    2015-03-01

    Full Text Available The purpose of this study was to investigate the effects of dietary addition of male and female Ferula eleaochytris powder (FEP on growth performance and body components of broilers (Ross-308. Treatment groups were allocated to 5 dietary in which 0 (control, 5 g and 10 g male FEP, 5 g and 10 g female FEP doses per kg commercial broiler diet. In total, 80 male broiler chicks (1-d old in which 16 animals in each group were used in study. Body weight gain and feed intake were monitored weekly for 6 weeks. Forty two days old broiler chicks were slaughtered to determine the possible changes in body components. The results of the study indicated that FEP had no effect on any parameters. However; the weight of the control group gave higher values than 5 g male Ferula group. In conclusion the measured parameters had no effect between all groups.

  17. The Two-Component Model for Calculating Total Body Fat from Body Density: An Evaluation in Healthy Women before, during and after Pregnancy

    Directory of Open Access Journals (Sweden)

    Elisabet Forsum

    2014-12-01

    Full Text Available A possibility to assess body composition during pregnancy is often important. Estimating body density (DB and use the two-component model (2CM to calculate total body fat (TBF represents an option. However, this approach has been insufficiently evaluated during pregnancy. We evaluated the 2CM, and estimated fat-free mass (FFM density and variability in 17 healthy women before pregnancy, in gestational weeks 14 and 32, and 2 weeks postpartum based on DB (underwater weighing, total body water (deuterium dilution and body weight, assessed on these four occasions. TBF, calculated using the 2CM and published FFM density (TBF2CM, was compared to reference estimates obtained using the three-component model (TBF3CM. TBF2CM minus TBF3CM (mean ± 2SD was −1.63 ± 5.67 (p = 0.031, −1.39 ± 7.75 (p = 0.16, −0.38 ± 4.44 (p = 0.49 and −1.39 ± 5.22 (p = 0.043 % before pregnancy, in gestational weeks 14 and 32 and 2 weeks postpartum, respectively. The effect of pregnancy on the variability of FFM density was larger in gestational week 14 than in gestational week 32. The 2CM, based on DB and published FFM density, assessed body composition as accurately in gestational week 32 as in non-pregnant adults. Corresponding values in gestational week 14 were slightly less accurate than those obtained before pregnancy.

  18. Chemical treatment of Escherichia coli. II. Direct extraction of recombinant protein from cytoplasmic inclusion bodies in intact cells.

    Science.gov (United States)

    Falconer, R J; O'Neill, B K; Middelberg, A P

    1998-02-20

    A method is presented for the direct extraction of the recombinant protein Long-R3-IGF-I from inclusion bodies located in the cytoplasm of intact Escherichia coli cells. Chemical treatment with 6M urea, 3 mM EDTA, and 20 mM dithiothreitol (DTT) at pH 9.0 proved an effective combination for extracting recombinant protein from intact cells. Comparable levels of Long-R3-IGF-I were recovered by direct extraction as achieved by in vitro dissolution following mechanical disruption. However, the purity of directly extracted recombinant protein was lower due to contamination by bacterial cell components. The kinetics of direct extraction are described using a first-order equation with the time constant of 3 min. Urea appears important for permeabilization of the cell and dissolution of the inclusion body. Conversely, EDTA is involved in permeabilization of the cell wall and DTT enhances protein release. pH proved to be important with lower levels of protein release achieved at low pH values (Long-R3-IGF-I release and caused an observable increase in viscosity. Advantages of the direct extraction method include its speed, simplicity, and efficiency at releasing product. PMID:10099214

  19. Argonaute2 Protein in Rat Spermatogenic Cells Is Localized to Nuage Structures and LAMP2-Positive Vesicles Surrounding Chromatoid Bodies.

    Science.gov (United States)

    Fujii, Yuki; Onohara, Yuko; Fujita, Hideaki; Yokota, Sadaki

    2016-04-01

    Localization of Argonaute2 (AGO2) protein-an essential component for the processing of small interfering RNA (siRNA)-directed RNA interference (RNAi) in RNA-induced silencing complex (RISC) in nuage of rat spermatogenic cells-was evaluated by immunofluorescence microscopy (IFM) and immunoelectron microscopy (IEM). AGO2 was shown, for the first time, to be localized to four previously classified types of nuage: irregularly shaped perinuclear granules (ISPGs), intermitochondrial cement (IMC), satellite bodies (SBs), and chromatoid bodies (CBs). Dual IEM staining for AGO2/Maelstrom (MAEL) protein or AGO2/MIWI protein demonstrated that AGO2 is colocalized with MAEL or MIWI proteins in these types of nuage. Dual IFM and IEM staining of AGO2/lysosomal-associated membrane protein 2 (LAMP2) showed that CB in round spermatids are in contact with and surrounded by LAMP2-positive vesicles, whereas nuage in pachytene spermatocytes are not. Taken together, our findings indicate that: (i) AGO2 in pachytene spermatocytes functions in ISPGs, IMC, and SBs; (ii) AGO2 in round spermatids functions in CBs, and that CBs are associated with lysosomal compartments. PMID:27029769

  20. Body protein losses estimated by nitrogen balance and potassium-40 counting

    International Nuclear Information System (INIS)

    Body protein losses estimated from N balance were compared with those estimated by 40K counting. Six nonlactating dairy cows were fed an adequate N diet for 7 wk, a low N diet for 9 wk, and a replete N diet for 3 wk. The low N diet contained high cell wall grass hay plus ground corn, starch, and molasses. Soybean meal was added to the low N diet to increase N in the adequate N and replete N diets. Intake was measured daily. Digestibilities, N balance, and body composition (estimated by 40K counting) were determined during each dietary regimen. During low N treatment, hay dry matter intake declined 2 kg/d, and supplement increased about .5 kg/d. Dry matter digestibility was not altered by N treatment. Protein and acid detergent fiber digestibilities decreased from 40 and 36% during adequate N to 20 and 2%, respectively, during low N. Fecal and urinary N also declined when cows were fed the low N diet. By the end of repletion, total intake, fiber, and protein digestibilities as well as N partition were similar to or exceeded those during adequate N intake. Body protein (N) loss was estimated by N balance to be about 3 kg compared with 8 kg by 40K counting. Body fat losses (32 kg) were large because of low energy digestibility and intake. Seven kilograms of body fat were regained during repletion, but there was no change in body protein

  1. A two-component system regulates gene expression of the type IX secretion component proteins via an ECF sigma factor.

    Science.gov (United States)

    Kadowaki, Tomoko; Yukitake, Hideharu; Naito, Mariko; Sato, Keiko; Kikuchi, Yuichiro; Kondo, Yoshio; Shoji, Mikio; Nakayama, Koji

    2016-01-01

    The periodontopathogen Porphyromonas gingivalis secretes potent pathogenic proteases, gingipains, via the type IX secretion system (T9SS). This system comprises at least 11 components; however, the regulatory mechanism of their expression has not yet been elucidated. Here, we found that the PorY (PGN_2001)-PorX (PGN_1019)-SigP (PGN_0274) cascade is involved in the regulation of T9SS. Surface plasmon resonance (SPR) analysis revealed a direct interaction between a recombinant PorY (rPorY) and a recombinant PorX (rPorX). rPorY autophosphorylated and transferred a phosphoryl group to rPorX in the presence of Mn(2+). These results demonstrate that PorX and PorY act as a response regulator and a histidine kinase, respectively, of a two component system (TCS), although they are separately encoded on the chromosome. T9SS component-encoding genes were down-regulated in a mutant deficient in a putative extracytoplasmic function (ECF) sigma factor, PGN_0274 (SigP), similar to the porX mutant. Electrophoretic gel shift assays showed that rSigP bound to the putative promoter regions of T9SS component-encoding genes. The SigP protein was lacking in the porX mutant. Co-immunoprecipitation and SPR analysis revealed the direct interaction between SigP and PorX. Together, these results indicate that the PorXY TCS regulates T9SS-mediated protein secretion via the SigP ECF sigma factor. PMID:26996145

  2. A two-component system regulates gene expression of the type IX secretion component proteins via an ECF sigma factor

    Science.gov (United States)

    Kadowaki, Tomoko; Yukitake, Hideharu; Naito, Mariko; Sato, Keiko; Kikuchi, Yuichiro; Kondo, Yoshio; Shoji, Mikio; Nakayama, Koji

    2016-01-01

    The periodontopathogen Porphyromonas gingivalis secretes potent pathogenic proteases, gingipains, via the type IX secretion system (T9SS). This system comprises at least 11 components; however, the regulatory mechanism of their expression has not yet been elucidated. Here, we found that the PorY (PGN_2001)-PorX (PGN_1019)-SigP (PGN_0274) cascade is involved in the regulation of T9SS. Surface plasmon resonance (SPR) analysis revealed a direct interaction between a recombinant PorY (rPorY) and a recombinant PorX (rPorX). rPorY autophosphorylated and transferred a phosphoryl group to rPorX in the presence of Mn2+. These results demonstrate that PorX and PorY act as a response regulator and a histidine kinase, respectively, of a two component system (TCS), although they are separately encoded on the chromosome. T9SS component-encoding genes were down-regulated in a mutant deficient in a putative extracytoplasmic function (ECF) sigma factor, PGN_0274 (SigP), similar to the porX mutant. Electrophoretic gel shift assays showed that rSigP bound to the putative promoter regions of T9SS component-encoding genes. The SigP protein was lacking in the porX mutant. Co-immunoprecipitation and SPR analysis revealed the direct interaction between SigP and PorX. Together, these results indicate that the PorXY TCS regulates T9SS-mediated protein secretion via the SigP ECF sigma factor. PMID:26996145

  3. Male pheromone protein components activate female vomeronasal neurons in the salamander Plethodon shermani

    Directory of Open Access Journals (Sweden)

    Feldhoff Pamela W

    2006-03-01

    Full Text Available Abstract Background The mental gland pheromone of male Plethodon salamanders contains two main protein components: a 22 kDa protein named Plethodon Receptivity Factor (PRF and a 7 kDa protein named Plethodon Modulating Factor (PMF, respectively. Each protein component individually has opposing effects on female courtship behavior, with PRF shortening and PMF lengthening courtship. In this study, we test the hypothesis that PRF or PMF individually activate vomeronasal neurons. The agmatine-uptake technique was used to visualize chemosensory neurons that were activated by each protein component individually. Results Vomeronasal neurons exposed to agmatine in saline did not demonstrate significant labeling. However, a population of vomeronasal neurons was labeled following exposure to either PRF or PMF. When expressed as a percent of control level labeled cells, PRF labeled more neurons than did PMF. These percentages for PRF and PMF, added together, parallel the percentage of labeled vomeronasal neurons when females are exposed to the whole pheromone. Conclusion This study suggests that two specific populations of female vomeronasal neurons are responsible for responding to each of the two components of the male pheromone mixture. These two neural populations, therefore, could express different receptors which, in turn, transmit different information to the brain, thus accounting for the different female behavior elicited by each pheromone component.

  4. Effect of protein source and resistance training on body composition and sex hormones

    Directory of Open Access Journals (Sweden)

    Krieger Diane R

    2007-07-01

    Full Text Available Abstract Background Evidence suggests an inverse relationship between soy protein intake and serum concentrations of male sex hormones. Anecdotal evidence indicates that these alterations in serum sex hormones may attenuate changes in lean body mass following resistance training. However, little empirical data exists regarding the effects of soy and milk-based proteins on circulating androgens and exercise induced body composition changes. Methods For 12 weeks 20 subjects were supplemented with 50 g per day of one of four different protein sources (Soy concentrate; Soy isolate; Soy isolate and whey blend, and Whey blend only in combination with a resistance-training program. Body composition, testosterone, estradiol and sex hormone binding globulin (SHBG were measured at baseline and week 12. Results Protein supplementation resulted in a significant increase in lean body mass independent of protein source (0.5 ± 1.1 and 0.9 ± 1.4 kg, p = 0.006, p = 0.007. No significant differences were observed between groups for total and free testosterone, SHBG, percentage body fat, BMI or body weight. The Testosterone/Estradiol ratio increased across all groups (+13.4, p = 0.005 and estradiol decreased (p = 0.002. Within group analysis showed significant increases in the Testosterone/Estradiol ratio in soy isolate + whey blend group (+16.3, p = 0.030. Estradiol was significantly lower in the whey blend group (-9.1 ± 8.7 pg/ml, p = 0.033. Conclusion This investigation shows that 12 week supplementation with soy protein does not decrease serum testosterone or inhibit lean body mass changes in subjects engaged in a resistance exercise program.

  5. Impaired Lysosomal Integral Membrane Protein 2-dependent Peroxiredoxin 6 Delivery to Lamellar Bodies Accounts for Altered Alveolar Phospholipid Content in Adaptor Protein-3-deficient pearl Mice.

    Science.gov (United States)

    Kook, Seunghyi; Wang, Ping; Young, Lisa R; Schwake, Michael; Saftig, Paul; Weng, Xialian; Meng, Ying; Neculai, Dante; Marks, Michael S; Gonzales, Linda; Beers, Michael F; Guttentag, Susan

    2016-04-15

    The Hermansky Pudlak syndromes (HPS) constitute a family of disorders characterized by oculocutaneous albinism and bleeding diathesis, often associated with lethal lung fibrosis. HPS results from mutations in genes of membrane trafficking complexes that facilitate delivery of cargo to lysosome-related organelles. Among the affected lysosome-related organelles are lamellar bodies (LB) within alveolar type 2 cells (AT2) in which surfactant components are assembled, modified, and stored. AT2 from HPS patients and mouse models of HPS exhibit enlarged LB with increased phospholipid content, but the mechanism underlying these defects is unknown. We now show that AT2 in the pearl mouse model of HPS type 2 lacking the adaptor protein 3 complex (AP-3) fails to accumulate the soluble enzyme peroxiredoxin 6 (PRDX6) in LB. This defect reflects impaired AP-3-dependent trafficking of PRDX6 to LB, because pearl mouse AT2 cells harbor a normal total PRDX6 content. AP-3-dependent targeting of PRDX6 to LB requires the transmembrane protein LIMP-2/SCARB2, a known AP-3-dependent cargo protein that functions as a carrier for lysosomal proteins in other cell types. Depletion of LB PRDX6 in AP-3- or LIMP-2/SCARB2-deficient mice correlates with phospholipid accumulation in lamellar bodies and with defective intraluminal degradation of LB disaturated phosphatidylcholine. Furthermore, AP-3-dependent LB targeting is facilitated by protein/protein interaction between LIMP-2/SCARB2 and PRDX6 in vitro and in vivo Our data provide the first evidence for an AP-3-dependent cargo protein required for the maturation of LB in AT2 and suggest that the loss of PRDX6 activity contributes to the pathogenic changes in LB phospholipid homeostasis found HPS2 patients. PMID:26907692

  6. Effects of Two Different Levels of Dietary Protein on Body Composition and Protein Nutritional Status of Growing Rats

    OpenAIRE

    Marcelo Macedo Rogero; Sandra Maria Lima Ribeiro; Julio Tirapegui; Ivanir Santana de Oliveira Pires

    2012-01-01

    This study aimed to investigate the effect of a high-protein diet on growth, body composition, and protein nutritional status of young rats. Newly-weaned Wistar rats, weighing 45–50 g, were distributed in two experimental groups, according to their diets, which contained 12% (G12) or 26% protein (G26), over a period of 3 weeks. The animals were euthanized at the end of this period and the following analyses were performed: chemical composition of the carcass, proteoglycan synthesis, IGF-I con...

  7. In vitro biosynthesis of globular proteins by murine splenic lymphocytes: effect of serum components as supplement

    International Nuclear Information System (INIS)

    Studies on replacement of foetal calf serum (FCS) with precipitable protein, non precipitable protein, dialysable and non dialysable components of the FCS in media for the growth and proliferation of murine splenic rat lymphocytes have revealed that the whole serum could be completely replaced by either of the components without any appreciable deleterious effect on the mitogenic response but none of these components could offer optimum immune response. These findings establish a covalent association of whole FCS for synthesis and secretion of immunologically important pulsed proteins in terms of turnover rate and quantification by FPLC and suggest an important and yet undefined role of FCS in the process of immunoglobulin synthesis. (author). 10 refs., 2 figs., 1 tab

  8. Localization of Seed Oil Body Proteins in Tobacco Protoplasts Reveals Specific Mechanisms of Protein Targeting to Leaf Lipid Droplets

    Institute of Scientific and Technical Information of China (English)

    Stefania De Domenico; Stefania Bonsegna; Marcello Salvatore Lenucci; Palmiro Poltronieri; Gian Pietro Di Sansebastiano; Angelo Santino

    2011-01-01

    Oleosin,caleosin and steroleosin are normally expressed in developing seed cells and are targeted to oil bodies.In the present work,the cDNA of each gene tagged with fluorescent proteins was transiently expressed into tobacco protoplasts and the fluorescent patterns observed by confocal laser scanning microscopy.Our results indicated clear differences in the endocellular localization of the three proteins.Oleosin and caleosin both share a common structure consisting of a central hydrophobic domain flanked by two hydrophilic domains and were correctly targeted to lipid droplets (LD),whereas steroleosin,characterized by an N-terminal oil body anchoring domain,was mainly retained in the endoplasmic reticulum (ER).Protoplast fractionation on sucrose gradients indicated that both oleosin and caleosingreen fluorescent protein (GFP) peaked at different fractions than where steroleosin-GFP or the ER marker binding immunoglobulin protein (BiP),were recovered.Chemical analysis confirmed the presence of triacylglycerols in one of the fractions where oleosin-GFP was recovered.Finally,only oleosinand caleosin-GFP were able to reconstitute artificial oil bodies in the presence of triacylglycerols and phospholipids.Taken together,our results pointed out for the first time that leaf LDs can be separated by the ER and both oleosin or caleosin are selectively targeted due to the existence of selective mechanisms controlling protein association with these organelles.

  9. Component Specificity for the Thylakoidal Sec and Delta Ph–Dependent Protein Transport Pathways

    OpenAIRE

    Mori, Hiroki; Summer, Elizabeth J.; Ma, Xianyue; Cline, Kenneth

    1999-01-01

    Prokaryotes and prokaryote-derived thylakoid membranes of chloroplasts share multiple, evolutionarily conserved pathways for protein export. These include the Sec, signal recognition particle (SRP), and Delta pH/Tat systems. Little is known regarding the thylakoid membrane components involved in these pathways. We isolated a cDNA clone to a novel component of the Delta pH pathway, Tha4, and prepared antibodies against pea Tha4, against maize Hcf106, a protein implicated in Delta pH pathway tr...

  10. Outer membrane protein binding sites of complement component 3 during opsonization of Haemophilus influenzae.

    OpenAIRE

    Hetherington, S V; Patrick, C C; Hansen, E J

    1993-01-01

    Complement component 3 (C3) binding to Haemophilus influenzae type b (Hib) is an important step in host defense against invasive disease, but the details of this process remain poorly understood. We have shown that the P1 and P2 outer membrane proteins (OMPs) serve as binding sites for C3 on serum-opsonized Hib. Whole-cell lysates of opsonized Hib were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the resolved proteins were transferred to nitrocellulose. Immunobl...

  11. Cosuppression of the alpha subunits of beta-conglycinin in transgenic soybean seeds induces the formation of endoplasmic reticulum-derived protein bodies.

    Science.gov (United States)

    Kinney, A J; Jung, R; Herman, E M

    2001-05-01

    The expression of the alpha and alpha' subunits of beta-conglycinin was suppressed by sequence-mediated gene silencing in transgenic soybean seed. The resulting seeds had similar total oil and protein content and ratio compared with the parent line. The decrease in beta-conglycinin protein was apparently compensated by an increased accumulation of glycinin. In addition, proglycinin, the precursor of glycinin, was detected as a prominent polypeptide band in the protein profile of the transgenic seed extract. Electron microscopic analysis and immunocytochemistry of maturing transgenic soybean seeds indicated that the process of storage protein accumulation was altered in the transgenic line. In normal soybeans, the storage proteins are deposited in pre-existing vacuoles by Golgi-derived vesicles. In contrast, in transgenic seed with reduced beta-conglycinin levels, endoplasmic reticulum (ER)-derived vesicles were observed that resembled precursor accumulating-vesicles of pumpkin seeds and the protein bodies accumulated by cereal seeds. Their ER-derived membrane of the novel vesicles did not contain the protein storage vacuole tonoplast-specific protein alpha-TIP, and the sequestered polypeptides did not contain complex glycans, indicating a preGolgi and nonvacuolar nature. Glycinin was identified as a major component of these novel protein bodies and its diversion from normal storage protein trafficking appears to be related to the proglycinin buildup in the transgenic seed. The stable accumulation of proteins in a protein body compartment instead of vacuolar accumulation of proteins may provide an alternative intracellular site to sequester proteins when soybeans are used as protein factories. PMID:11340189

  12. A human telomerase holoenzyme protein required for Cajal body localization and telomere synthesis

    OpenAIRE

    Venteicher, Andrew S.; Abreu, Eladio B.; Meng, Zhaojing; McCann, Kelly E.; Terns, Rebecca M.; Veenstra, Timothy D.; Terns, Michael P.; Artandi, Steven E.

    2009-01-01

    Telomerase is a ribonucleoprotein (RNP) complex that synthesizes telomere repeats in tissue progenitor cells and cancer cells. Active human telomerase consists of at least three principal subunits, including the telomerase reverse transcriptase (TERT), the telomerase RNA (TERC), and dyskerin. Here, we identify a holoenzyme subunit, TCAB1 (telomerase Cajal body protein1), uniquely enriched in Cajal bodies, nuclear sites of RNP processing important for telomerase function. TCAB1 associates with...

  13. A super-ductile alloy for the die-casting of aluminium automotive body structural components

    OpenAIRE

    Watson, D.; Ji, S; Fan, Z.

    2014-01-01

    Super-ductile die-cast aluminium alloys are critical to future light-weighting of automotive body structures. This paper introduces a die-cast aluminium alloy that can satisfy the requirements of these applications. After a review of currently available alloys, the requirement of a die-cast aluminium alloy for automotive body structural parts is proposed and an Al-Mg-Si system is suggested. The effect of the alloying elements, in the composition, has been investigated on the microstructure an...

  14. Three-body recombination of two-component cold atomic gases into deep dimers in an optical model

    DEFF Research Database (Denmark)

    Mikkelsen, Mathias; Jensen, A. S.; Fedorov, D. V.; Zinner, Nikolaj Thomas

    2015-01-01

    We consider three-body recombination into deep dimers in a mass-imbalanced two-component atomic gas. We use an optical model where a phenomenological imaginary potential is added to the lowest adiabatic hyper-spherical potential. The consequent imaginary part of the energy eigenvalue corresponds to...... the decay rate or recombination probability of the three-body system. The method is formulated in details and the relevant qualitative features are discussed as functions of scattering lengths and masses. We use zero-range model in analyses of recent recombination data. The dominating scattering...... length is usually related to the non-equal two-body systems. We account for temperature smearing which tends to wipe out the higher-lying Efimov peaks. The range and the strength of the imaginary potential determine positions and shapes of the Efimov peaks as well as the absolute value of the...

  15. Optimization of physical training of students of high school with regard to quantitative features muscular components of their bodies

    Directory of Open Access Journals (Sweden)

    Kolokoltsev M.M.

    2015-02-01

    Full Text Available Purpose : to provide a quantitative description of the muscle component of students’ body with regard to their motor characteristics to improve training in the discipline "Physical Education". Material : a study of muscular component of the body in 1937 students aged 17-20 years old of age living in the Baikal region. Motor quality students were evaluated by tests. Problem analysis was conducted based on the data of Polish authors. Results : the dependence of the amount of content in muscle mass in the body of the frequency of physical training in high school. Also found significantly higher levels of performance in motor tests in the group of students with a high level of expression of muscle mass. Conclusions : the studies have shown a direct relationship content of lean body mass of locomotor activity. Set better indicator values in tests of physical fitness with a high content of muscle tissue. Lack of exercise training on older years is recommended to compensate for self-manage motor activities.

  16. The effect of psoriasis treatment on body composition, components of metabolic syndrome and psoriatic arthritis

    Directory of Open Access Journals (Sweden)

    Funda Tamer

    2015-03-01

    Full Text Available Background and Design: Psoriasis is a chronic inflammatory immun mediated skin disorder with unknown etiology. The chronic inflammation in psoriasis have role in the development of metabolic and vascular disorders related with associating comorbidities. Recent studies have suggested a strong association exists between metabolic syndrome, obesity and complexity of the association between psoriasis, body mass index (BMI and psoriasis tratment. In this study, our aim was to investigate the effect of psoriasis treatment with methotrexate, cyclosporine and biological agents on body composition, comorbidities and associated laboratory findings. Materials and Methods: Seventy-nine patients treated with methotrexate, cyclosporin and biological agents were included in our study. Demographic characteristics, body composition analysis, psoriasis related comorbidities and laboratory examinations were evaluated before and after 12 weeks of systemic treatment. Results: Comorbidities and metabolic syndrome tended to be more frequent in the anti tumor necrosis factor alpha (anti-TNF-α treated group. Increase in body fat and weight detected in patiens receiving biologic drug therapy. Conclusion: The results of our study showed that severe psoriasis patients with longer disease duration were more likely to have metabolic syndrome because of severe and long term inflammation in pathogenesis of comorbidities.

  17. Winter body mass and over-ocean flocking as components of danger management by Pacific dunlins

    NARCIS (Netherlands)

    Ydenberg, R.C.; Dekker, D.; Kaiser, G.; Shepherd, P.C.F.; Ogden, L.E.; Rickards, K.; Lank, D.B.

    2010-01-01

    Background: We compared records of the body mass and roosting behavior of Pacific dunlins (Calidris alpina pacifica) wintering on the Fraser River estuary in southwest British Columbia between the 1970s and the 1990s. 'Over-ocean flocking' is a relatively safe but energetically-expensive alternative

  18. Autoantibodies to protein transport and messenger RNA processing pathways: endosomes, lysosomes, Golgi complex, proteasomes, assemblyosomes, exosomes, and GW bodies.

    Science.gov (United States)

    Stinton, Laura M; Eystathioy, Theophany; Selak, Sanja; Chan, Edward K L; Fritzler, Marvin J

    2004-01-01

    Over 50 years ago the lupus erythematosus (LE) cell phenomenon was described and this was quickly followed by the introduction of the LE cell test and indirect immunofluorescence (IIF) to detect antinuclear antibodies (ANA) in clinical laboratories. Recently, attention has turned to the identification of the autoantigens that bind to cytoplasmic organelles such as the Golgi complex, endosomes and other "cytoplasmic somes". Three endosome autoantigens include early endosome antigen 1 (EEA1, 160 kDa), cytoplasmic linker protein-170 (CLIP-170, 170 kDa), and lysobisphosphatidic acid (LBPA). Antibodies to EEA1 were seen in a variety of conditions but approximately 40% of the patients had a neurological disease. Despite the prominence of lysosomes in cells and tissues, reports of autoantibodies are limited to the lysosomal antigen h-LAMP-2 and the cytoplasmic antineutrophil antibodies (cANCA). Autoantigens in the Golgi complex include giantin/macrogolgin, golgin-245, golgin 160, golgin-97, golgin 95/gm130, and golgin-67. More recently, there has been an interest in autoantibodies that bind components of the "SMN complex" or the "assemblyosome". Arginine/glycine (RG)-rich domains in components of the SMN complex interact with Sm, like-Sm (LSm), fibrillarin, RNA helicase A (Gu), and coilin proteins, all of which are antigen targets in a variety of diseases. More recently, components of a novel cytoplasmic structure named GW bodies (GWBs) have been identified as targets of human autoantibodies. Components of GWBs include GW182, a unique mRNA-binding protein, like Sm proteins (LSms), and decapping (hDcp1) and exonuclease (Xrn) enzymes. Current evidence suggests that GWBs are involved in the cytoplasmic processing of mRNAs. Autoantibodies to the "cytoplasmic somes" are relatively uncommon and serological tests to detect most of them are not widely available. PMID:14962794

  19. Winter body mass and over-ocean flocking as components of danger management by Pacific dunlins

    Directory of Open Access Journals (Sweden)

    Ogden Lesley

    2010-01-01

    Full Text Available Abstract Background We compared records of the body mass and roosting behavior of Pacific dunlins (Calidris alpina pacifica wintering on the Fraser River estuary in southwest British Columbia between the 1970s and the 1990s. 'Over-ocean flocking' is a relatively safe but energetically-expensive alternative to roosting during the high tide period. Fat stores offer protection against starvation, but are a liability in escape performance, and increase flight costs. Peregrine falcons (Falco peregrinus were scarce on the Fraser River estuary in the 1970s, but their numbers have since recovered, and they prey heavily on dunlins. The increase has altered the balance between predation and starvation risks for dunlins, and thus how dunlins regulate roosting behavior and body mass to manage the danger. We therefore predicted an increase in the frequency of over-ocean flocking as well as a decrease in the amount of fat carried by dunlins over these decades. Results Historical observations indicate that over-ocean flocking of dunlins was rare prior to the mid-1990s and became common thereafter. Residual body masses of dunlins were higher in the 1970s, with the greatest difference between the decades coinciding with peak peregrine abundance in October, and shrinking over the course of winter as falcon seasonal abundance declines. Whole-body fat content of dunlins was lower in the 1990s, and accounted for most of the change in body mass. Conclusions Pacific dunlins appear to manage danger in a complex manner that involves adjustments both in fat reserves and roosting behavior. We discuss reasons why over-ocean flocking has apparently become more common on the Fraser estuary than at other dunlin wintering sites.

  20. Circadian proteins CLOCK and BMAL1 in the chromatoid body, a RNA processing granule of male germ cells.

    Directory of Open Access Journals (Sweden)

    Rita L Peruquetti

    Full Text Available Spermatogenesis is a complex differentiation process that involves genetic and epigenetic regulation, sophisticated hormonal control, and extensive structural changes in male germ cells. RNA nuclear and cytoplasmic bodies appear to be critical for the progress of spermatogenesis. The chromatoid body (CB is a cytoplasmic organelle playing an important role in RNA post-transcriptional and translation regulation during the late steps of germ cell differentiation. The CB is also important for fertility determination since mutations of genes encoding its components cause infertility by spermatogenesis arrest. Targeted ablation of the Bmal1 and Clock genes, which encode central regulators of the circadian clock also result in fertility defects caused by problems other than spermatogenesis alterations. We show that the circadian proteins CLOCK and BMAL1 are localized in the CB in a stage-specific manner of germ cells. Both BMAL1 and CLOCK proteins physically interact with the ATP-dependent DEAD-box RNA helicase MVH (mouse VASA homolog, a hallmark component of the CB. BMAL1 is differentially expressed during the spermatogenic cycle of seminiferous tubules, and Bmal1 and Clock deficient mice display significant CB morphological alterations due to BMAL1 ablation or low expression. These findings suggest that both BMAL1 and CLOCK contribute to CB assembly and physiology, raising questions on the role of the circadian clock in reproduction and on the molecular function that CLOCK and BMAL1 could potentially have in the CB assembly and physiology.

  1. Purifying selection drives the evolution of surfactant protein C (SP-C) independently of body temperature regulation in mammals.

    Science.gov (United States)

    Potter, Sally; Orgeig, Sandra; Donnellan, Stephen; Daniels, Christopher B

    2007-06-01

    The pulmonary surfactant system of heterothermic mammals must be capable of dealing with the effect of low body temperatures on the physical state of the lipid components. We have shown previously that there is a modest increase in surfactant cholesterol during periods of torpor, however these changes do not fully explain the capacity of surfactant to function under the wide range of physical conditions imposed by torpor. Here we examine indirectly the role of surfactant protein C (SP-C) in adapting to variable body temperatures by testing for the presence of positive (adaptive) selection during evolutionary transitions between heterothermy and homeothermy. We sequenced SP-C from genomic DNA of 32 mammalian species from groups of closely related heterothermic and homeothermic species (contrasts). We used phylogenetic analysis by maximum likelihood estimates of rates of non-synonymous to synonymous substitutions and fully Bayesian inference of these sequences to determine whether the mode of body temperature regulation exerts a selection pressure driving the molecular adaptation of SP-C. The protein sequence of SP-C is highly conserved with synonymous or highly conservative amino acid substitutions being predominant. The evolution of SP-C among mammals is characterised by high codon usage bias and high rates of transition/transversion. The only contrast to show evidence of positive selection was that of the bears (Ursus americanus and U. maritimus). The significance of this result is unclear. We show that SP-C is under strong evolutionary constraints, driven by purifying selection, presumably to maintain protein function despite variation in the mode of body temperature regulation. PMID:20483290

  2. The Role of Cgrp-Receptor Component Protein (Rcp in Cgrp-Mediated Signal Transduction

    Directory of Open Access Journals (Sweden)

    M. A. Prado

    2001-01-01

    Full Text Available The calcitonin gene-related peptide (CGRP-receptor component protein (RCP is a 17-kDa intracellular peripheral membrane protein required for signal transduction at CGRP receptors. To determine the role of RCP in CGRP-mediated signal transduction, RCP was depleted from NIH3T3 cells using antisense strategy. Loss of RCP protein correlated with loss of cAMP production by CGRP in the antisense cells. In contrast, loss of RCP had no effect on CGRP-mediated binding; therefore RCP is not acting as a chaperone for the CGRP receptor. Instead, RCP is a novel signal transduction molecule that couples the CGRP receptor to the cellular signal transduction machinery. RCP thus represents a prototype for a new class of signal transduction proteins that are required for regulation of G protein-coupled receptors.

  3. Ephemeral protein binding to DNA shapes stable nuclear bodies and chromatin domains

    CERN Document Server

    Brackley, C A; Michieletto, D; Mouvet, F; Cook, P R; Marenduzzo, D

    2016-01-01

    Fluorescence microscopy reveals that the contents of many (membrane-free) nuclear "bodies" exchange rapidly with the soluble pool whilst the underlying structure persists; such observations await a satisfactory biophysical explanation. To shed light on this, we perform large-scale Brownian dynamics simulations of a chromatin fiber interacting with an ensemble of (multivalent) DNA-binding proteins; these proteins switch between two states -- active (binding) and inactive (non-binding). This system provides a model for any DNA-binding protein that can be modified post-translationally to change its affinity for DNA (e.g., like the phosphorylation of a transcription factor). Due to this out-of-equilibrium process, proteins spontaneously assemble into clusters of self-limiting size, as individual proteins in a cluster exchange with the soluble pool with kinetics like those seen in photo-bleaching experiments. This behavior contrasts sharply with that exhibited by "equilibrium", or non-switching, proteins that exis...

  4. Steady-state dynamics of Cajal body components in the Xenopus germinal vesicle

    OpenAIRE

    Handwerger, Korie E.; Murphy, Christine; Gall, Joseph G.

    2003-01-01

    Cajal bodies (CBs) are evolutionarily conserved nuclear organelles that contain many factors involved in the transcription and processing of RNA. It has been suggested that macromolecular complexes preassemble or undergo maturation within CBs before they function elsewhere in the nucleus. Most such models of CB function predict a continuous flow of molecules between CBs and the nucleoplasm, but there are few data that directly support this view. We used fluorescence recovery after photobleach...

  5. The nuclear pore complex–associated protein, Mlp2p, binds to the yeast spindle pole body and promotes its efficient assembly

    OpenAIRE

    Niepel, Mario; Strambio-De-Castillia, Caterina; Fasolo, Joseph; Chait, Brian T.; Rout, Michael P

    2005-01-01

    The two yeast proteins Mlp1p and Mlp2p (homologues of the vertebrate protein Tpr) are filamentous proteins attached to the nuclear face of nuclear pore complexes. Here we perform a proteomic analysis, which reveals that the two Mlps have strikingly different interacting partners, testifying to their different roles within the cell. We find that Mlp2p binds directly to Spc110p, Spc42p, and Spc29p, which are three core components of the spindle pole body (SPB), the nuclear envelope–associated y...

  6. The herpesvirus associated ubiquitin specific protease, USP7, is a negative regulator of PML proteins and PML nuclear bodies.

    Directory of Open Access Journals (Sweden)

    Feroz Sarkari

    Full Text Available The PML tumor suppressor is the founding component of the multiprotein nuclear structures known as PML nuclear bodies (PML-NBs, which control several cellular functions including apoptosis and antiviral effects. The ubiquitin specific protease USP7 (also called HAUSP is known to associate with PML-NBs and to be a tight binding partner of two herpesvirus proteins that disrupt PML NBs. Here we investigated whether USP7 itself regulates PML-NBs. Silencing of USP7 was found to increase the number of PML-NBs, to increase the levels of PML protein and to inhibit PML polyubiquitylation in nasopharyngeal carcinoma cells. This effect of USP7 was independent of p53 as PML loss was observed in p53-null cells. PML-NBs disruption was induced by USP7 overexpression independently of its catalytic activity and was induced by either of the protein interaction domains of USP7, each of which localized to PML-NBs. USP7 also disrupted NBs formed from some single PML isoforms, most notably isoforms I and IV. CK2α and RNF4, which are known regulators of PML, were dispensable for USP7-associated PML-NB disruption. The results are consistent with a novel model of PML regulation where a deubiquitylase disrupts PML-NBs through recruitment of another cellular protein(s to PML NBs, independently of its catalytic activity.

  7. The herpesvirus associated ubiquitin specific protease, USP7, is a negative regulator of PML proteins and PML nuclear bodies.

    Science.gov (United States)

    Sarkari, Feroz; Wang, Xueqi; Nguyen, Tin; Frappier, Lori

    2011-01-01

    The PML tumor suppressor is the founding component of the multiprotein nuclear structures known as PML nuclear bodies (PML-NBs), which control several cellular functions including apoptosis and antiviral effects. The ubiquitin specific protease USP7 (also called HAUSP) is known to associate with PML-NBs and to be a tight binding partner of two herpesvirus proteins that disrupt PML NBs. Here we investigated whether USP7 itself regulates PML-NBs. Silencing of USP7 was found to increase the number of PML-NBs, to increase the levels of PML protein and to inhibit PML polyubiquitylation in nasopharyngeal carcinoma cells. This effect of USP7 was independent of p53 as PML loss was observed in p53-null cells. PML-NBs disruption was induced by USP7 overexpression independently of its catalytic activity and was induced by either of the protein interaction domains of USP7, each of which localized to PML-NBs. USP7 also disrupted NBs formed from some single PML isoforms, most notably isoforms I and IV. CK2α and RNF4, which are known regulators of PML, were dispensable for USP7-associated PML-NB disruption. The results are consistent with a novel model of PML regulation where a deubiquitylase disrupts PML-NBs through recruitment of another cellular protein(s) to PML NBs, independently of its catalytic activity. PMID:21305000

  8. A component modes projection and assembly model reduction methodology for articulated, multi-flexible body structures

    Science.gov (United States)

    Lee, Allan Y.; Tsuha, Walter S.

    1993-01-01

    A two-stage model reduction methodology, combining the classical Component Mode Synthesis (CMS) method and the newly developed Enhanced Projection and Assembly (EP&A) method, is proposed in this research. The first stage of this methodology, called the COmponent Modes Projection and Assembly model REduction (COMPARE) method, involves the generation of CMS mode sets, such as the MacNeal-Rubin mode sets. These mode sets are then used to reduce the order of each component model in the Rayleigh-Ritz sense. The resultant component models are then combined to generate reduced-order system models at various system configurations. A composite mode set which retains important system modes at all system configurations is then selected from these reduced-order system models. In the second stage, the EP&A model reduction method is employed to reduce further the order of the system model generated in the first stage. The effectiveness of the COMPARE methodology has been successfully demonstrated on a high-order, finite-element model of the cruise-configured Galileo spacecraft.

  9. Shape distortion and thermo-mechanical properties of SOFC components from green tape to sintering body

    DEFF Research Database (Denmark)

    Teocoli, Francesca; Ni, De Wei; Tadesse Molla, Tesfaye;

    due to binder burn out, differential shrinkage behavior and to a potential interfacial reaction between the two materials. To analyze the phenomena, shrinkage of SOFC components single layers and bilayered samples were measured insitu by optical dilatometer. The densification mismatch stress, due...

  10. Snf7p, a Component of the ESCRT-III Protein Complex, Is an Upstream Member of the RIM101 Pathway in Candida albicans

    OpenAIRE

    Kullas, Amy L.; Li, Mingchun; Davis, Dana A.

    2004-01-01

    The success of Candida albicans as an opportunistic pathogen is based in part on its ability to adapt to diverse environments. The RIM101 pathway governs adaptation to neutral-alkaline environments and is required for virulence. Analysis of a genomic two-hybrid study conducted with Saccharomyces cerevisiae revealed that components involved in multivesicular bodies (MVB) transport may interact with RIM101 pathway members. Thus, we hypothesized that these proteins may function in the RIM101 pat...

  11. Control of foreign polypeptide localization in specific layers of protein body type I in rice seed

    OpenAIRE

    Sasou, Ai; Shigemitsu, Takanari; Saito, Yuhi; Tanaka, Manami; Morita, Shigeto; Masumura, Takehiro

    2016-01-01

    Key message Prolamin–GFP fusion proteins, expressed under the control of native prolamin promoters, were localized in specific layers of PB-Is. Prolamin–GFP fusion proteins were gradually digested from outside by pepsin digestion. Abstract In rice seed endosperm, protein body type I (PB-I) has a layered structure consisting of prolamin species and is the resistant to digestive juices in the intestinal tract. We propose the utilization of PB-Is as an oral vaccine carrier to induce mucosal immu...

  12. Allometric Growth Patterns of Body and Carcass Components in Ardhi Goat

    OpenAIRE

    A. Al-Owaimer; Suliman, G.; A. El-Waziry; Metwally, H.; M. Abouheif

    2013-01-01

    This study aims to evaluate the developmental trends and the allometric growth values of various body parts and fat depots of the most prevailing indigenous Saudi goat. Thirty male Ardhi kids were serially slaughtered at 10, 15, 20, 25, 30 and 35 kg live weight. As the kids grew, the bones of hind limb grew at slower rates than the bones of the forelimb and within each limb, the cannon bone grew relatively at a slower rate than the upper skeletal bones. The allometric coefficients for the gro...

  13. Protein Degradation and Protection Observed in the Presence of Novel Wound Dressing Components

    Directory of Open Access Journals (Sweden)

    Laura K.S. Parnell

    2011-12-01

    Full Text Available Chronic wounds typically have excessive levels of matrix metalloproteinases (MMPs and proinflammatory cytokines that impair healing. Reducing these detrimental proteins may be key to healing chronic wounds. Proprietary protease blends were formulated specifically to degrade excessive amounts of proinflammatory factors that could prevent wound healing. Applications of protease-containing wound dressings to acute and chronic wounds have been observed clinically to resolve inflammation and appear to aid healing. The purpose of this study was to test in vitro a deliberate blend of proteases for the ability to deactivate or activate known proteins associated with inflammation or healing. Purified human target proteins were incubated with test and control solutions and samples removed at various time points. Blinded samples were tested using a novel infrared protein multiplex sandwich-ELISA-type array technique. Many proinflammatory proteins such as MMPs, cytokines and chemokines were susceptible to degradation. Many proteins such as growth factors, cytokines and TIMP1 were resistant to degradation. Not all proinflammatory proteins were deactivated. Family protein structure did not appear to affect susceptibility to degradation or deactivation. These results suggest that specific protease containing wound dressings appear to reduce multiple detrimental components which may disrupt their deleterious effects on the wound bed and microenvironment. By improving the wound microenvironment through the use of definitive proteases, these novel wound dressings may help transition wounds into the subsequent phase of healing.

  14. Comparative study of contents of several bioactive components in fruiting bodies and mycelia of culinary-medicinal mushrooms.

    Science.gov (United States)

    Lin, Shin-Yi; Chen, Yu-Kai; Yu, Hui-Tzu; Barseghyan, Gayane S; Asatiani, Mikheil D; Wasser, Solomon P; Mau, Jeng-Leun

    2013-01-01

    Mushrooms (including fruiting bodies and mycelia) contain several bioactive components such as lovastatin, γ-aminobutyric acid (GABA), and ergothioneine. This article reports the results of 49 samples, including 9 fruiting bodies, 39 mycelia, and 1 vegetative cell, of 35 species of culinary-medicinal mushrooms from 18 genera: Agaricus, Agrocybe, Coprinus, Cordyceps, Cyathus, Daedalia, Flammulina, Fomes, Ganoderma, Grifola, Laetiporus, Lentinus, Morchella, Ophiocordyceps, Pleurotus, Trametes, Tremella, and Verpa. The results show that Cyathus striatus strain 978 contained the highest amount of lovastatin (995.66 mg/kg) in mycelia. Among fruiting bodies, 6 samples contained a high amount of GABA (274.86-822.45 mg/kg), whereas among mycelia, contents of GABA in 27 samples ranged from 215.36 to 2811.85 mg/kg. Among mycelia, Pleurotus cornucopiae strain 1101 contained the highest amount of ergothioneine (3482.09 mg/kg). Overall, these 3 bioactive components were commonly found in most mushrooms, and the results obtained might be related to their beneficial effects. PMID:23662618

  15. Allometric Growth Patterns of Body and Carcass Components in Ardhi Goat

    Directory of Open Access Journals (Sweden)

    A. Al-Owaimer

    2013-10-01

    Full Text Available This study aims to evaluate the developmental trends and the allometric growth values of various body parts and fat depots of the most prevailing indigenous Saudi goat. Thirty male Ardhi kids were serially slaughtered at 10, 15, 20, 25, 30 and 35 kg live weight. As the kids grew, the bones of hind limb grew at slower rates than the bones of the forelimb and within each limb, the cannon bone grew relatively at a slower rate than the upper skeletal bones. The allometric coefficients for the growth of hot and cold carcass, liver, stomach compartments and lean relative to empty body weight were isogonic (b = 1.00, whereas coefficients of all internal fat depots, intermuscular and subcutaneous fat weights were heterogonic with the high growth impetus of b values greater than 1.00 (p<0.01. The developmental rates of the intestines and separated bones from cold carcass side were heterogonic with medium growth impetus. These results, showed that the highest growth coefficients were obtained for omental and perirenal fat indicating the late maturing characteristics of these depots, followed in a decreasing order by mesenteric and intermuscular fat, channel fat and finally subcutaneous and pericardial fat, which were the earliest developing depots.

  16. Highly Efficient Production of Soluble Proteins from Insoluble Inclusion Bodies by a Two-Step-Denaturing and Refolding Method

    OpenAIRE

    Yang, Zhong; Zhang, Linlin; Zhang, Yan; Zhang, Ting; Feng, Yanye; Lu, Xiuxiu; Lan, Wenxian; Wang, Jufang; Wu, Houming; Cao, Chunyang; Wang, Xiaoning

    2011-01-01

    The production of recombinant proteins in a large scale is important for protein functional and structural studies, particularly by using Escherichia coli over-expression systems; however, approximate 70% of recombinant proteins are over-expressed as insoluble inclusion bodies. Here we presented an efficient method for generating soluble proteins from inclusion bodies by using two steps of denaturation and one step of refolding. We first demonstrated the advantages of this method over a conve...

  17. Individual whey protein components influence lipid oxidation dependent on pH

    DEFF Research Database (Denmark)

    Horn, Anna Frisenfeldt; Nielsen, Nina Skall; Jacobsen, Charlotte

    In emulsions, lipid oxidation is expected to be initiated at the oil-water interface. The properties of the emulsifier used and the composition at the interface is therefore expected to be of great importance for the resulting oxidation. Previous studies have shown that individual whey protein...... affecting the preferential adsorption of whey protein components at the interface. The aim of the study was to compare lipid oxidation in 10% fish oil-in-water emulsions prepared with 1% whey protein having either a high concentration of α-lactalbumin, a high concentration of β-lactoglobulin or equal...... amounts of the two. Emulsions were prepared at pH4 and pH7. Emulsions were characterized by their droplet sizes, viscosities, and contents of proteins in the water phase. Lipid oxidation was assessed by PV and secondary volatile oxidation products. Results showed that pH greatly influenced the oxidative...

  18. The Tanita SC-240 to Assess Body Composition in Pre-School Children: An Evaluation against the Three Component Model

    Directory of Open Access Journals (Sweden)

    Christine Delisle Nyström

    2016-06-01

    Full Text Available Quick, easy-to-use, and valid body composition measurement options for young children are needed. Therefore, we evaluated the ability of the bioelectrical impedance (BIA device, Tanita SC-240, to measure fat mass (FM, fat free mass (FFM and body fatness (BF% in 40 healthy, Swedish 5.5 years old children against the three component model (3C model. Average BF%, FM, and FFM for BIA were: 19.4% ± 3.9%, 4.1 ± 1.9 kg, and 16.4 ± 2.4 kg and were all significantly different (p < 0.001 from corresponding values for the 3C model (25.1% ± 5.5%, 5.3 ± 2.5 kg, and 15.2 ± 2.0 kg. Bland and Altman plots had wide limits of agreement for all body composition variables. Significant correlations ranging from 0.81 to 0.96 (p < 0.001 were found for BF%, FM, and FFM between BIA and the 3C model. When dividing the children into tertiles for BF%, 60% of children were classified correctly by means of BIA. In conclusion, the Tanita SC-240 underestimated BF% in comparison to the 3C model and had wide limits of agreement. Further work is needed in order to find accurate and easy-to-use methods for assessing body composition in pre-school children.

  19. The Tanita SC-240 to Assess Body Composition in Pre-School Children: An Evaluation against the Three Component Model

    Science.gov (United States)

    Delisle Nyström, Christine; Henriksson, Pontus; Alexandrou, Christina; Löf, Marie

    2016-01-01

    Quick, easy-to-use, and valid body composition measurement options for young children are needed. Therefore, we evaluated the ability of the bioelectrical impedance (BIA) device, Tanita SC-240, to measure fat mass (FM), fat free mass (FFM) and body fatness (BF%) in 40 healthy, Swedish 5.5 years old children against the three component model (3C model). Average BF%, FM, and FFM for BIA were: 19.4% ± 3.9%, 4.1 ± 1.9 kg, and 16.4 ± 2.4 kg and were all significantly different (p < 0.001) from corresponding values for the 3C model (25.1% ± 5.5%, 5.3 ± 2.5 kg, and 15.2 ± 2.0 kg). Bland and Altman plots had wide limits of agreement for all body composition variables. Significant correlations ranging from 0.81 to 0.96 (p < 0.001) were found for BF%, FM, and FFM between BIA and the 3C model. When dividing the children into tertiles for BF%, 60% of children were classified correctly by means of BIA. In conclusion, the Tanita SC-240 underestimated BF% in comparison to the 3C model and had wide limits of agreement. Further work is needed in order to find accurate and easy-to-use methods for assessing body composition in pre-school children. PMID:27322313

  20. Dynamics of the component of body composition in athletes playing sports with damage to the medial meniscus of the knee

    Directory of Open Access Journals (Sweden)

    Moh'd Khalil Moh'd Abdel Kader

    2012-11-01

    Full Text Available The question of the influence of the developed program of physical rehabilitation with the use of modern means and methods of recovery on the performance component of body composition in athletes playing sports after arthroscopic stapling the medial meniscus was considered. The analysis and synthesis of scientific and methodological data on the determination of body composition analysis techniques using bioelectrical resistance was shown. Found that long-term immobilization that accompanies the process of rehabilitation, reduced physical activity, vascular and other changes lead to muscle atrophy, restriction of mobility in the knee joint, trophic changes of the injured limb. The use of the physical rehabilitation programs allowed to state its high efficiency.

  1. Refolding in high hydrostatic pressure of recombinant proteins from inclusion bodies in Escherichia Coli

    International Nuclear Information System (INIS)

    The expression of proteins as inclusion bodies in bacteria is a widely used alternative for production of recombinant protein. However, the aggregation is a problem often encountered during refolding of these proteins. High hydrostatic pressure are able to solubilise the inclusion bodies in the presence of low concentrations of denaturant reagents, encouraging refolding protein with high efficiency and reduce costs. This work aims to refolding of recombinant proteins expressed in Escherichia coli from inclusion bodies using high hydrostatic pressure. Three toxins, all featuring five or more disulfide bonds were studied: NXH8, Natterin 2 and Bothropstoxin 1. Suspensions of inclusion bodies of the three proteins were pressurized to 2000 bars for 16 hours. The buffers were optimized for refolding of the three proteins. The buffer used in the refolding of NXH8 was 50 mM Tris HCl, pH 9.0 with proportion of 1GSH: 4GSSG at a concentration of 6 mM and 2 M GdnHCl. Inclusion bodies were used in O.D. (A600nm) of 0.5. After refolding process, dialysis was performed at pH 7.0. The final yield of obtaining soluble NXH8 was 40% (28,6 mg of soluble NXH8/L of culture medium). The refolding of Bothropstoxin 1 was obtained in refolding buffer of Tris HCl 50 mM, pH 7,5 with proportion of 2 GSH: GSSG 3 and concentration of 3 mM and 1 M GdnHCl. Use with a suspension of O.D. (A600nm) of 0.5. The final yield of recovery of Bothropstoxin 1 refolded was 32% (9,2 mg of refolded Bothropstoxin 1/L of culture medium). The refolding of Natterin 2 was performed in the refolding buffer: 20 mM Tris HCl pH 9.0 at a ratio of 2 GSH: 3GSSG and concentration of 10 mM and 1 M GdnHCl and inclusion bodies O.D. (A600nm) of 6.0. The yield of Natterin 2 refolded was 20% (3,7 mg/L of culture medium). Physico-chemical and biological analysis were performed by SDS-PAGE, western blot, scanning electron microscopy, biological tests in vivo and in vitro and structural. The analysis conducted in NXH8 did not show

  2. Poly (ADP-ribose polymerase 1 is required for protein localization to Cajal body.

    Directory of Open Access Journals (Sweden)

    Elena Kotova

    2009-02-01

    Full Text Available Recently, the nuclear protein known as Poly (ADP-ribose Polymerase1 (PARP1 was shown to play a key role in regulating transcription of a number of genes and controlling the nuclear sub-organelle nucleolus. PARP1 enzyme is known to catalyze the transfer of ADP-ribose to a variety of nuclear proteins. At present, however, while we do know that the main acceptor for pADPr in vivo is PARP1 protein itself, by PARP1 automodification, the significance of PARP1 automodification for in vivo processes is not clear. Therefore, we investigated the roles of PARP1 auto ADP-ribosylation in dynamic nuclear processes during development. Specifically, we discovered that PARP1 automodification is required for shuttling key proteins into Cajal body (CB by protein non-covalent interaction with pADPr in vivo. We hypothesize that PARP1 protein shuttling follows a chain of events whereby, first, most unmodified PARP1 protein molecules bind to chromatin and accumulate in nucleoli, but then, second, upon automodification with poly(ADP-ribose, PARP1 interacts non-covalently with a number of nuclear proteins such that the resulting protein-pADPr complex dissociates from chromatin into CB.

  3. Evaluation of two novel leptospiral proteins for their interaction with human host components.

    Science.gov (United States)

    Silva, Lucas P; Fernandes, Luis G V; Vieira, Monica L; de Souza, Gisele O; Heinemann, Marcos B; Vasconcellos, Silvio A; Romero, Eliete C; Nascimento, Ana L T O

    2016-07-01

    Pathogenic species of the genus Leptospira are the etiological agents of leptospirosis, the most widespread zoonosis. Mechanisms involved in leptospiral pathogenesis are not well understood. By data mining the genome sequences of Leptospira interrogans we have identified two proteins predicted to be surface exposed, LIC10821 and LIC10064. Immunofluorescence and proteinase K assays confirmed that the proteins are exposed. Reactivity of the recombinant proteins with human sera has shown that rLIC10821, but not rLIC10064, is recognized by antibodies in confirmed leptospirosis serum samples, suggesting its expression during infection. The rLIC10821 was able to bind laminin, in a dose-dependent fashion, and was called Lsa37 (leptospiral surface adhesin of 37 kDa). Studies with human plasma components demonstrated that rLIC10821 interacts with plasminogen (PLG) and fibrinogen (Fg). The binding of Lsa37 with PLG generates plasmin when PLG activator was added. Fibrin clotting reduction was observed in a thrombin-catalyzed reaction, when Fg was incubated with Lsa37, suggesting that this protein may interfere in the coagulation cascade during the disease. Although LIC10064 protein is more abundant than the corresponding Lsa37, binding activity with all the components tested was not detected. Thus, Lsa37 is a novel versatile adhesin that may mediate Leptospira-host interactions. PMID:27129366

  4. Tapasin-related protein TAPBPR is an additional component of the MHC class I presentation pathway

    OpenAIRE

    Boyle, Louise H.; Hermann, Clemens; Boname, Jessica M.; Porter, Keith M; Patel, Peysh A.; Burr, Marian L; Duncan, Lidia M.; Harbour, Michael E.; Rhodes, David A.; Skjødt, Karsten; Lehner, Paul J.; Trowsdale, John

    2013-01-01

    Tapasin is an integral component of the peptide-loading complex (PLC) important for efficient peptide loading onto MHC class I molecules. We investigated the function of the tapasin-related protein, TAPBPR. Like tapasin, TAPBPR is widely expressed, IFN-γ–inducible, and binds to MHC class I coupled with β2-microglobulin in the endoplasmic reticulum. In contrast to tapasin, TAPBPR does not bind ERp57 or calreticulin and is not an integral component of the PLC. β2-microglobulin is essential for ...

  5. Study of Differentiation and Characteristics of Sieve Elements and P-protein Body in Ginkgo biloba

    Institute of Scientific and Technical Information of China (English)

    LING Yuping; CHEN Peng; LI Li; WANG Lili; PAN Bin; TAI Yunlin

    2006-01-01

    The ultrastructure observation, energy spectrum analyses and study on sieve elements in gingko (Ginkgo biloba. L) root phloem showed: 1) The inside wall had a large retention of small grains with a diameter of 3- 4 um, and the outside wall membrane structure wrapped P-protein body with a kernel of high electronic density. 2) The growth had the typical process of programmed cell death (PCD), i.e. the cell karyoplasm condensed, chromatins congealed and contracted and gradually agglomerated to the periphery of the nuclear membrane, and accordingly a nuclear of malformation appeared; the endoplasmic reticulum extended and connected and merged mutually, and the shallow part fused with the cell membrane. After packing the cell organelle, the endoplasmic reticulum divided the cell into several combined necrotic corpuscles of different sizes, the cell organelles collapsing with the cell and gathering abundantly in the inside cell wall. There were two kinds of degradation for the mitochondria: One was that the electronic density of the matrix decreased, the ridges died out to be cavitations gradually, and the double-layer membrane broke and ruined; the other was that parts of the territorial structure of the membrane broke, and the substance it contained escaped. 3) The ultrastructure of the course of gingko root phloem developing from cell with protein thin wall into sieve elements might make out that the P-protein body was produced in PCD and developed with retention in the combined necrotic corpuscle. 4) The P-protein had higher contents of S, K, P than the sieve elements wall tissue, and the percentage composition of S was 4.64%, which was more than twice the composition in the cell wall with 2.14 %. According to the observation through transmission electron microscope that showed the P-protein had high electronic density, we might affirm that it was a P-protein body with S, P. The composition of K was 21.62%, 1.52 times that of the cell wall which was 14.23%. The

  6. Human papillomavirus (HPV) type 16 E7 protein bodies cause tumour regression in mice

    International Nuclear Information System (INIS)

    Human papillomaviruses (HPV) are the causative agents of cervical cancer in women, which results in over 250 000 deaths per year. Presently there are two prophylactic vaccines on the market, protecting against the two most common high-risk HPV types 16 and 18. These vaccines remain very expensive and are not generally affordable in developing countries where they are needed most. Additionally, there remains a need to treat women that are already infected with HPV, and who have high-grade lesions or cervical cancer. In this paper, we characterize the immunogenicity of a therapeutic vaccine that targets the E7 protein of the most prevalent high-risk HPV - type 16 – the gene which has previously been shown to be effective in DNA vaccine trials in mice. The synthetic shuffled HPV-16 E7 (16E7SH) has lost its transforming properties but retains all naturally-occurring CTL epitopes. This was genetically fused to Zera®, a self-assembly domain of the maize γ-zein able to induce the accumulation of recombinant proteins into protein bodies (PBs), within the endoplasmic reticulum in a number of expression systems. High-level expression of the HPV 16E7SH protein fused to Zera® in plants was achieved, and the protein bodies could be easily and cost-effectively purified. Immune responses comparable to the 16E7SH DNA vaccine were demonstrated in the murine model, with the protein vaccine successfully inducing a specific humoral as well as cell mediated immune response, and mediating tumour regression. The fusion of 16E7SH to the Zera® peptide was found to enhance the immune responses, presumably by means of a more efficient antigen presentation via the protein bodies. Interestingly, simply mixing the free PBs and 16E7SH also enhanced immune responses, indicating an adjuvant activity for the Zera® PBs

  7. Fish meals, fish components, and fish protein hydrolysates as potential ingredients in pet foods.

    Science.gov (United States)

    Folador, J F; Karr-Lilienthal, L K; Parsons, C M; Bauer, L L; Utterback, P L; Schasteen, C S; Bechtel, P J; Fahey, G C

    2006-10-01

    An experiment to determine the chemical composition and protein quality of 13 fish substrates (pollock by-products, n = 5; fish protein hydrolysates, n = 5; and fish meals, n = 3) was conducted. Two of these substrates, salmon protein hydrolysate (SPH) and salmon meal with crushed bones (SMB), were used to determine their palatability as components of dog diets. Pollock by-products differed in concentrations of CP, crude fat, and total AA by 71, 79, and 71%, respectively, and GE by 4.1 kcal/g. Fish protein hydrolysates and fish meals were less variable (approximately 18, 14, and 17%, and 1.4 kcal/g, respectively). Biogenic amine concentrations were much higher in fish protein hydrolysates as compared with pollock by-products and fish meals. Pollock liver and viscera had the highest total fatty acid concentrations; however, red salmon hydrolysate and SMB had the highest total PUFA concentrations (49.63 and 48.60 mg/g, respectively). Salmon protein hydrolysate had the highest protein solubility in 0.2% KOH. Based on calculations using immobilized digestive enzyme assay values, lysine digestibility of fish meal substrates was comparable to in vivo cecectomized rooster assay values and averaged approximately 90.3%. Also, pollock milt, pollock viscera, red salmon hydrolysate, and sole hydrolysate had comparable values as assessed by immobilized digestive enzyme assay and rooster assays. A chick protein efficiency ratio (PER) assay compared SMB and SPH to a whole egg meal control and showed that SMB had high protein quality (PER = 3.5), whereas SPH had poor protein quality (PER value less than 1.5). However, using whole egg meal as the reference protein, both fish substrates were found to be good protein sources with an essential AA index of 1.0 and 0.9 for SMB and SPH, respectively. In the dog palatability experiments, a chicken-based control diet and 2 diets containing 10% of either SPH or SMB were tested. Dogs consumed more of the SPH diet compared with the control

  8. IQGAP1 is a novel CXCR2-interacting protein and essential component of the "chemosynapse".

    Directory of Open Access Journals (Sweden)

    Nicole F Neel

    Full Text Available Chemotaxis is essential for a number of physiological processes including leukocyte recruitment. Chemokines initiate intracellular signaling pathways necessary for chemotaxis through binding seven transmembrane G protein-couple receptors. Little is known about the proteins that interact with the intracellular domains of chemokine receptors to initiate cellular signaling upon ligand binding. CXCR2 is a major chemokine receptor expressed on several cell types, including endothelial cells and neutrophils. We hypothesize that multiple proteins interact with the intracellular domains of CXCR2 upon ligand stimulation and these interactions comprise a "chemosynapse", and play important roles in transducing CXCR2 mediated signaling processes.In an effort to define the complex of proteins that assemble upon CXCR2 activation to relay signals from activated chemokine receptors, a proteomics approach was employed to identify proteins that co-associate with CXCR2 with or without ligand stimulation. The components of the CXCR2 "chemosynapse" are involved in processes ranging from intracellular trafficking to cytoskeletal modification. IQ motif containing GTPase activating protein 1 (IQGAP1 was among the novel proteins identified to interact directly with CXCR2. Herein, we demonstrate that CXCR2 co-localizes with IQGAP1 at the leading edge of polarized human neutrophils and CXCR2 expressing differentiated HL-60 cells. Moreover, amino acids 1-160 of IQGAP1 directly interact with the carboxyl-terminal domain of CXCR2 and stimulation with CXCL8 enhances IQGAP1 association with Cdc42.Our studies indicate that IQGAP1 is a novel essential component of the CXCR2 "chemosynapse".

  9. Application of principal component analysis in protein unfolding: An all-atom molecular dynamics simulation study

    Science.gov (United States)

    Das, Atanu; Mukhopadhyay, Chaitali

    2007-10-01

    We have performed molecular dynamics (MD) simulation of the thermal denaturation of one protein and one peptide—ubiquitin and melittin. To identify the correlation in dynamics among various secondary structural fragments and also the individual contribution of different residues towards thermal unfolding, principal component analysis method was applied in order to give a new insight to protein dynamics by analyzing the contribution of coefficients of principal components. The cross-correlation matrix obtained from MD simulation trajectory provided important information regarding the anisotropy of backbone dynamics that leads to unfolding. Unfolding of ubiquitin was found to be a three-state process, while that of melittin, though smaller and mostly helical, is more complicated.

  10. Counterregulation of insulin by leptin as key component of autonomic regulation of body weight

    Institute of Scientific and Technical Information of China (English)

    Katarina; T; Borer

    2014-01-01

    A re-examination of the mechanism controlling eating, locomotion, and metabolism prompts formulation of a new explanatory model containing five features: a coordinating joint role of the(1) autonomic nervous system(ANS);(2) the suprachiasmatic(SCN) master clock in counterbalancing parasympathetic digestive and absorptive functions and feeding with sympathetic locomotor and thermogenic energy expenditure within a circadian framework;(3) interaction of the ANS/SCN command with brain substrates of reward encompassing dopaminergic projections to ventral striatum and limbic and cortical forebrain. These drive the nonhomeostatic feeding and locomotor motivated behaviors in interaction with circulating ghrelin and lateral hypothalamic neurons signaling through melanin concentrating hormone and orexin-hypocretin peptides;(4) counterregulation of insulin by leptin of both gastric and adipose tissue origin through: potentiation by leptin of cholecystokinin-mediated satiation, inhibition of insulin secretion, suppression of insulin lipogenesis by leptin lipolysis, and modulation of peripheral tissue and brain sensitivity to insulin action. Thus weight-loss induced hypoleptimia raises insulin sensitivity and promotes its parasympathetic anabolic actions while obesity-induced hyperleptinemia supresses insulin lipogenic action; and(5) inhibition by leptin of bone mineral accrual suggesting that leptin may contribute to the maintenance of stability of skeletal, lean-body, as well as adipose tissue masses.

  11. Individual whey protein components influence lipid oxidation dependent on pH

    OpenAIRE

    Horn, Anna Frisenfeldt; Nielsen, Nina Skall; Jacobsen, Charlotte

    2012-01-01

    In emulsions, lipid oxidation is expected to be initiated at the oil-water interface. The properties of the emulsifier used and the composition at the interface is therefore expected to be of great importance for the resulting oxidation. Previous studies have shown that individual whey protein components (α-lactalbumin and β-lactoglobulin) adsorb differently to the interface depending on pH. In addition, differences has been shown to exists between the oxidative stability provided by α-lactal...

  12. Human CHMP6, a myristoylated ESCRT-III protein, interacts directly with an ESCRT-II component EAP20 and regulates endosomal cargo sorting.

    Science.gov (United States)

    Yorikawa, Chiharu; Shibata, Hideki; Waguri, Satoshi; Hatta, Kazumi; Horii, Mio; Katoh, Keiichi; Kobayashi, Toshihide; Uchiyama, Yasuo; Maki, Masatoshi

    2005-04-01

    CHMP6 (charged multivesicular body protein 6) is a human orthologue of yeast Vps (vacuolar protein sorting) 20, a component of ESCRT (endosomal sorting complex required for transport)-III. Various CHMP6 orthologues in organisms ranging from yeast to humans contain the N-myristoylation consensus sequence at each N-terminus. Metabolic labelling of HEK-293 (human embryonic kidney) cells showed the incorporation of [3H]myristate into CHMP6 fused C-terminally to GFP (green fluorescent protein) (CHMP6-GFP). Interactions of CHMP6 with another ESCRT-III component CHMP4b/Shax [Snf7 (sucrose non-fermenting 7) homologue associated with Alix] 1, one of three paralogues of human Vps32/Snf7, and with EAP20 (ELL-associated protein 20), a human counterpart of yeast Vps25 and component of ESCRT-II, were observed by co-immunoprecipitation of epitope-tagged proteins expressed in HEK-293 cells. The in vitro pull-down assays using their recombinant proteins purified from Escherichia coli demonstrated direct physical interactions which were mediated by the N-terminal basic half of CHMP6. Overexpressed CHMP6-GFP in HeLa cells exhibited a punctate distribution throughout the cytoplasm especially in the perinuclear area, as revealed by fluorescence microscopic analysis. Accumulation of LBPA (lysobisphosphatidic acid), a major phospholipid in internal vesicles of an MVB (multivesicular body), was observed in the CHMP6-GFP-localizing area. FLAG-tagged EAP20 distributed diffusely, but exhibited a punctate distribution on co-expression with CHMP6-GFP. Overexpression of CHMP6-GFP caused reduction of transferrin receptors on the plasma membrane surface, but caused their accumulation in the cytoplasm. Ubiquitinated proteins and endocytosed EGF continuously accumulated in CHMP6-GFP-expressing cells. These results suggest that CHMP6 acts as an acceptor for ESCRT-II on endosomal membranes and regulates cargo sorting. PMID:15511219

  13. Intraflagellar transport (IFT protein IFT25 is a phosphoprotein component of IFT complex B and physically interacts with IFT27 in Chlamydomonas.

    Directory of Open Access Journals (Sweden)

    Zhaohui Wang

    Full Text Available BACKGROUND: Intraflagellar transport (IFT is the bidirectional movement of IFT particles between the cell body and the distal tip of a flagellum. Organized into complexes A and B, IFT particles are composed of at least 18 proteins. The function of IFT proteins in flagellar assembly has been extensively investigated. However, much less is known about the molecular mechanism of how IFT is regulated. METHODOLOGY/PRINCIPAL FINDINGS: We herein report the identification of a novel IFT particle protein, IFT25, in Chlamydomonas. Dephosphorylation assay revealed that IFT25 is a phosphoprotein. Biochemical analysis of temperature sensitive IFT mutants indicated that IFT25 is an IFT complex B subunit. In vitro binding assay confirmed that IFT25 binds to IFT27, a Rab-like small GTPase component of the IFT complex B. Immunofluorescence staining showed that IFT25 has a punctuate flagellar distribution as expected for an IFT protein, but displays a unique distribution pattern at the flagellar base. IFT25 co-localizes with IFT27 at the distal-most portion of basal bodies, probably the transition zones, and concentrates in the basal body region by partially overlapping with other IFT complex B subunits, such as IFT46. Sucrose density gradient centrifugation analysis demonstrated that, in flagella, the majority of IFT27 and IFT25 including both phosphorylated and non-phosphorylated forms are cosedimented with other complex B subunits in the 16S fractions. In contrast, in cell body, only a fraction of IFT25 and IFT27 is integrated into the preassembled complex B, and IFT25 detected in complex B is preferentially phosphorylated. CONCLUSION/SIGNIFICANCE: IFT25 is a phosphoprotein component of IFT particle complex B. IFT25 directly interacts with IFT27, and these two proteins likely form a subcomplex in vivo. We postulate that the association and disassociation between the subcomplex of IFT25 and IFT27 and complex B might be involved in the regulation of IFT.

  14. Accurate design of megadalton-scale two-component icosahedral protein complexes.

    Science.gov (United States)

    Bale, Jacob B; Gonen, Shane; Liu, Yuxi; Sheffler, William; Ellis, Daniel; Thomas, Chantz; Cascio, Duilio; Yeates, Todd O; Gonen, Tamir; King, Neil P; Baker, David

    2016-07-22

    Nature provides many examples of self- and co-assembling protein-based molecular machines, including icosahedral protein cages that serve as scaffolds, enzymes, and compartments for essential biochemical reactions and icosahedral virus capsids, which encapsidate and protect viral genomes and mediate entry into host cells. Inspired by these natural materials, we report the computational design and experimental characterization of co-assembling, two-component, 120-subunit icosahedral protein nanostructures with molecular weights (1.8 to 2.8 megadaltons) and dimensions (24 to 40 nanometers in diameter) comparable to those of small viral capsids. Electron microscopy, small-angle x-ray scattering, and x-ray crystallography show that 10 designs spanning three distinct icosahedral architectures form materials closely matching the design models. In vitro assembly of icosahedral complexes from independently purified components occurs rapidly, at rates comparable to those of viral capsids, and enables controlled packaging of molecular cargo through charge complementarity. The ability to design megadalton-scale materials with atomic-level accuracy and controllable assembly opens the door to a new generation of genetically programmable protein-based molecular machines. PMID:27463675

  15. Crystallographic characterization of a multidomain histidine protein kinase from an essential two-component regulatory system

    International Nuclear Information System (INIS)

    The multidomain cytoplasmic portion of the histidine protein kinase from an essential two-component signal transduction system has been crystallized and X-ray data have been collected to 2.8 Å resolution. YycGF is a highly conserved two-component signal transduction system that is specific to low-G+C Gram-positive bacteria, including many important human pathogens. It has been recognized as a crucial regulatory system for cell-wall metabolism. YycG, the histidine protein kinase of this system, is a multidomain transmembrane protein. The truncated cytoplasmic portion of YycG from Bacillus subtilis encompassing the PAS domain, the dimerization domain and the catalytic domain was expressed, purified and crystallized. X-ray data were collected to 2.8 Å resolution with a completeness of 98.2% and an overall Rmerge of 5.6%. The crystals belonged to space group P61 or P65, with unit-cell parameters a = 135.0, c = 133.0 Å. The selenomethionine-substituted version of the protein was crystallized and X-ray data were collected to 3.6 Å resolution for subsequent MAD phasing

  16. The action to brain protein component induced by internal contamination of enriched uranium

    International Nuclear Information System (INIS)

    Objective: To study the changes in main component of brain protein induced by internal contamination of enriched uranium in different level. Methods: With low-temperature and high-speed centrifugation, ion exchange chromatography and dialysis, the authors separated and purified brain homogenate and got β-NGF from SD from SD rat's brain. Then authors tested the extracted β-NGF by neurite outgrowth test of PC12 cells and confirmed that it has high biological activity. Results: Experiments showed that enriched uranium may lead to the reduction of the obtainable rate of β-NGF. It should be noted, that the reduction has a linear relation with enriched uranium dose. Meanwhile authors found that the action of enriched uranium may lead to cleavage of brain protein, thereby enlarge the range of molecular weight spectrum of main component and increase the quantity of protein band in SDS-PAGE. Conclusion: The radio-toxicological effects induced by enriched uranium were the cleavage of brain protein as well as the reduction of the obtainable rate of β-NGF

  17. Phloem RNA-binding proteins as potential components of the long-distance RNA transport system.

    Directory of Open Access Journals (Sweden)

    VICENTE ePALLAS

    2013-05-01

    Full Text Available RNA-binding proteins (RBPs govern a myriad of different essential processes in eukaryotic cells. Recent evidence reveals that apart from playing critical roles in RNA metabolism and RNA transport, RBPs perform a key function in plant adaption to various environmental conditions. Long distance RNA transport occurs in land plants through the phloem, a conducting tissue that integrates the wide range of signalling pathways required to regulate plant development and response to stress processes. The macromolecules in the phloem pathway vary greatly and include defence proteins, transcription factors, chaperones acting in long distance trafficking, and RNAs (mRNAs, siRNAs and miRNAs. How these RNA molecules translocate through the phloem is not well understood, but recent evidence indicates the presence of translocatable RNA-binding proteins in the phloem, which act as potential components of long distance RNA transport system. This review updates our knowledge on the characteristics and functions of RBPs present in the phloem.

  18. The choice of label and measurement technique in tracer studies of body protein metabolism in man

    International Nuclear Information System (INIS)

    The turnover of non-serum proteins in man has had limited study despite the physiological importance of maintaining the balance between synthesis and breakdown of body proteins. Body protein is usually considered as a single pool and breakdown rates are often measured by monitoring excreted label at intervals after pulse labelling with radioactive or 15N amino acids. No label has yet been used for measuring tissue protein breakdown in man which is free from the major problem of label re-utilization. All measurements of breakdown rates, eg. with 75Se-selenomethionine, 15N- or 14C-glycine, give rate constants which are too low. The heterogeneity of body proteins also means that an estimate of the weighted average breakdown rate can only be obtained after following the excretion of isotope for a long period, perhaps of the order of 3-4 half-lives which, for man, would be 100 days after labelling. We therefore use infusions with either 14C- or 15N-labelled amino acids to measure breakdown and synthesis rates: these values are less affected by problems of protein heterogeneity. Single injection techniques are subject to more error than constant infusions of label because of the difficulty of defining the precursor activity. 15N labelling need not be confined to essential amino acids if total protein rather than amino acid turnover is studied: the latter involves measurements of the labelled amino acid itself which is difficult with 15N because of the small amounts of free amino acid nitrogen available. Carbon labelling of non-essential amino acids is unsuitable for studies of protein turnover and the choice of the position of the label on the molecule is important when labelled essential amino acids are employed. Short-term changes in protein metabolism are evaluated better with amino acids with a small pool size; the equilibration time in the excretory bicarbonate pool is also shorter than in the urea pool so that 15N is less useful than carbon labelling. We now use C

  19. The Tanita SC-240 to Assess Body Composition in Pre-School Children: An Evaluation against the Three Component Model.

    Science.gov (United States)

    Delisle Nyström, Christine; Henriksson, Pontus; Alexandrou, Christina; Löf, Marie

    2016-01-01

    Quick, easy-to-use, and valid body composition measurement options for young children are needed. Therefore, we evaluated the ability of the bioelectrical impedance (BIA) device, Tanita SC-240, to measure fat mass (FM), fat free mass (FFM) and body fatness (BF%) in 40 healthy, Swedish 5.5 years old children against the three component model (3C model). Average BF%, FM, and FFM for BIA were: 19.4% ± 3.9%, 4.1 ± 1.9 kg, and 16.4 ± 2.4 kg and were all significantly different (p FFM between BIA and the 3C model. When dividing the children into tertiles for BF%, 60% of children were classified correctly by means of BIA. In conclusion, the Tanita SC-240 underestimated BF% in comparison to the 3C model and had wide limits of agreement. Further work is needed in order to find accurate and easy-to-use methods for assessing body composition in pre-school children. PMID:27322313

  20. Modification effects of physical activity and protein intake on heritability of body size and composition

    DEFF Research Database (Denmark)

    Silventoinen, Karri; Hasselbalch, Ann Louise; Lallukka, Tea;

    2009-01-01

    Mx statistical package (Virginia Institute for Psychiatric and Behavioral Genetics, Virginia Commonwealth University, Richmond, VA). RESULTS: High physical activity was associated with lower mean values, and a high proportion of protein in the diet was associated with higher mean BMI, waist...... circumference, and percentage body fat and a reduction in genetic and environmental variances. Genetic modification by physical activity was statistically significant for BMI (-0.18; 95% CI: -0.31, -0.05) and waist circumference (-0.14; 95% CI: -0.22, -0.05) in the merged data. A high proportion of protein in...

  1. Caleosins: Ca+2 binding proteins associated with oil-bodies

    DEFF Research Database (Denmark)

    Næsted, Henrik; Frandsen, Gitte Inselmann; Jauh, G.Y.;

    2000-01-01

    We have previously identified a rice gene encoding a 27 kDa protein with a single Ca2+-binding EF-hand and a putative membrane anchor. We report here similar genes termed caleosins, CLO, in other plants and fungi; they comprise a multigene family of at least five members in Arabidopsis (AtClo1...... expression in developing embryos and also in root tip cells. Antibodies raised against AtCLO1 were used to detect caleosin in cellular fractions of Arabidopsis and rapeseed. This indicated that caleosins are a novel class of lipid body proteins, which may also be associated with an ER subdomain....

  2. Bioactive protein-based nanofibers interact with intestinal biological components resulting in transepithelial permeation of a therapeutic protein.

    Science.gov (United States)

    Stephansen, Karen; García-Díaz, María; Jessen, Flemming; Chronakis, Ioannis S; Nielsen, Hanne Mørck

    2015-11-10

    Proteins originating from natural sources may constitute a novel type of material for use in drug delivery. However, thorough understanding of the behavior and effects of such a material when processed into a matrix together with a drug is crucial prior to further development into a drug product. In the present study the potential of using bioactive electrospun fish sarcoplasmic proteins (FSP) as a carrier matrix for small therapeutic proteins was demonstrated in relation to the interactions with biological components of the intestinal tract. The inherent structural and chemical properties of FSP as a biomaterial facilitated interactions with cells and enzymes found in the gastrointestinal tract and displayed excellent biocompatibility. More specifically, insulin was efficiently encapsulated into FSP fibers maintaining its conformation, and subsequent controlled release was obtained in simulated intestinal fluid. The encapsulation of insulin into FSP fibers provided protection against chymotrypsin degradation, and resulted in an increase in insulin transport to around 12% without compromising the cellular viability. This increased transport was driven by interactions upon contact between the nanofibers and the Caco-2 cell monolayer leading to the opening of the tight junction proteins. Overall, electrospun FSP may constitute a novel material for oral delivery of biopharmaceuticals. PMID:26320547

  3. Prevalence of metabolic syndrome and its components among Chinese professional athletes of strength sports with different body weight categories.

    Directory of Open Access Journals (Sweden)

    Jianjun Guo

    Full Text Available BACKGROUND: There is an increasing concern on cardiometabolic health in young professional athletes at heavy-weight class. OBJECTIVE: Our cross-sectional survey aimed to evaluate the prevalence of metabolic syndrome and clustering of metabolic risk factors in a population of young and active professional athletes of strength sports in China. METHODS: From July 2006 to December 2008, a total of 131 male and 130 female athletes of strength sports were enrolled. We used two criteria provided by the Chinese Diabetes Society (2004 and the National Cholesterol Education Program's Adult Treatment Panel III (2002 to define the metabolic syndrome and its individual components, respectively. RESULTS: Regardless of their similar ages (mean: 21 years and exercise levels, athletes in the heaviest-weight-class with unlimited maximum body weight (UBW boundaries (mean weight and BMI: 130 kg and 38 kg/m(2 for men, 110 kg and 37 kg/m(2 for women had significantly higher prevalence of metabolic syndrome than did those in all other body-weight-class with limited body weight (LBW boundaries (mean weight and BMI: 105 kg and 32 kg/m(2 for men, 70 kg and 26 kg/m(2 for women. Prevalence of metabolic syndrome using CDS criteria (UBW vs. LBW: 89% vs. 18% for men, 47% vs. 0% for women and its individual components, including central obesity, hypertension, hypertriglyceridemia, low high-density lipoprotein-cholesterol levels, and impaired fasting glucose, were all significantly higher in athletes at the heaviest weight group with UBW than all other weight groups with LBW. CONCLUSIONS: Our study suggests that professional athletes of strength sports at the heaviest-weight-class are at a significant increased risk of cardiometabolic disease compared with those at all other weight categories. The findings support the importance of developing and implementing the strategy of early screening, awareness, and interventions for weight-related health among young athletes.

  4. Effect of Protein Intake on Strength, Body Composition and Endocrine Changes in Strength/Power Athletes

    Directory of Open Access Journals (Sweden)

    Kang Jie

    2006-12-01

    Full Text Available Abstract Comparison of protein intakes on strength, body composition and hormonal changes were examined in 23 experienced collegiate strength/power athletes participating in a 12-week resistance training program. Subjects were stratified into three groups depending upon their daily consumption of protein; below recommended levels (BL; 1.0 – 1.4 g·kg-1·day-1; n = 8, recommended levels (RL; 1.6 – 1.8 g·kg-1·day-1; n = 7 and above recommended levels (AL; > 2.0 g·kg-1·day-1; n = 8. Subjects were assessed for strength [one-repetition maximum (1-RM bench press and squat] and body composition. Resting blood samples were analyzed for total testosterone, cortisol, growth hormone, and insulin-like growth factor. No differences were seen in energy intake (3,171 ± 577 kcal between the groups, and the energy intake for all groups were also below the recommended levels for strength/power athletes. No significant changes were seen in body mass, lean body mass or fat mass in any group. Significant improvements in 1-RM bench press and 1-RM squat were seen in all three groups, however no differences between the groups were observed. Subjects in AL experienced a 22% and 42% greater change in Δ 1-RM squat and Δ 1-RM bench press than subjects in RL, however these differences were not significant. No significant changes were seen in any of the resting hormonal concentrations. The results of this study do not provide support for protein intakes greater than recommended levels in collegiate strength/power athletes for body composition improvements, or alterations in resting hormonal concentrations.

  5. Searching the Evolutionary Origin of Epithelial Mucus Protein Components-Mucins and FCGBP.

    Science.gov (United States)

    Lang, Tiange; Klasson, Sofia; Larsson, Erik; Johansson, Malin E V; Hansson, Gunnar C; Samuelsson, Tore

    2016-08-01

    The gel-forming mucins are large glycosylated proteins that are essential components of the mucus layers covering epithelial cells. Using novel methods of identifying mucins based on profile hidden Markov models, we have found a large number of such proteins in Metazoa, aiding in their classification and allowing evolutionary studies. Most vertebrates have 5-6 gel-forming mucin genes and the genomic arrangement of these genes is well conserved throughout vertebrates. An exception is the frog Xenopus tropicalis with an expanded repertoire of at least 26 mucins of this type. Furthermore, we found that the ovomucin protein, originally identified in chicken, is characteristic of reptiles, birds, and amphibians. Muc6 is absent in teleost fish, but we now show that it is present in animals such as ghost sharks, demonstrating an early origin in vertebrate evolution. Public RNA-Seq data were analyzed with respect to mucins in zebrafish, frog, and chicken, thus allowing comparison in regard of tissue and developmental specificity. Analyses of invertebrate proteins reveal that gel-forming-mucin type of proteins is widely distributed also in this group. Their presence in Cnidaria, Porifera, and in Ctenophora (comb jellies) shows that these proteins were present early in metazoan evolution. Finally, we examined the evolution of the FCGBP protein, abundant in mucus and related to gel-forming mucins in terms of structure and localization. We demonstrate that FCGBP, ubiquitous in vertebrates, has a conserved N-terminal domain. Interestingly, this domain is also present as an N-terminal sequence in a number of bacterial proteins. PMID:27189557

  6. Comparative genomic analysis of two-component regulatory proteins in Pseudomonas syringae

    Directory of Open Access Journals (Sweden)

    Ussery David W

    2007-10-01

    Full Text Available Abstract Background Pseudomonas syringae is a widespread bacterial plant pathogen, and strains of P. syringae may be assigned to different pathovars based on host specificity among different plant species. The genomes of P. syringae pv. syringae (Psy B728a, pv. tomato (Pto DC3000 and pv. phaseolicola (Pph 1448A have been recently sequenced providing a major resource for comparative genomic analysis. A mechanism commonly found in bacteria for signal transduction is the two-component system (TCS, which typically consists of a sensor histidine kinase (HK and a response regulator (RR. P. syringae requires a complex array of TCS proteins to cope with diverse plant hosts, host responses, and environmental conditions. Results Based on the genomic data, pattern searches with Hidden Markov Model (HMM profiles have been used to identify putative HKs and RRs. The genomes of Psy B728a, Pto DC3000 and Pph 1448A were found to contain a large number of genes encoding TCS proteins, and a core of complete TCS proteins were shared between these genomes: 30 putative TCS clusters, 11 orphan HKs, 33 orphan RRs, and 16 hybrid HKs. A close analysis of the distribution of genes encoding TCS proteins revealed important differences in TCS proteins among the three P. syringae pathovars. Conclusion In this article we present a thorough analysis of the identification and distribution of TCS proteins among the sequenced genomes of P. syringae. We have identified differences in TCS proteins among the three P. syringae pathovars that may contribute to their diverse host ranges and association with plant hosts. The identification and analysis of the repertoire of TCS proteins in the genomes of P. syringae pathovars constitute a basis for future functional genomic studies of the signal transduction pathways in this important bacterial phytopathogen.

  7. Effectiveness of exercise and protein supplementation intervention on body composition, functional fitness, and oxidative stress among elderly Malays with sarcopenia.

    Science.gov (United States)

    Shahar, Suzana; Kamaruddin, Norshafarina Shari; Badrasawi, Manal; Sakian, Noor Ibrahim Mohamed; Abd Manaf, Zahara; Yassin, Zaitun; Joseph, Leonard

    2013-01-01

    Sarcopenia, characterized as muscle loss that occurs with aging, is a major health problem in an aging population, due to its implications on mobility, quality of life, and fall risk. Protein supplementation could improve the physical fitness by increasing protein anabolism, and exercise has a documented evidence of positive effect on functional status among the elderly. However, the combined effect of both protein supplementation and exercise has not been investigated among sarcopenic elderly in the Asian population. Thus, this study aimed to determine the effectiveness of exercise intervention and protein supplementation either alone or in combination for 12 weeks, on body composition, functional fitness, and oxidative stress among elderly Malays with sarcopenia. Sixty five sarcopenic elderly Malays aged 60-74 years were assigned to the control group, exercise group (ExG), protein supplementation group (PrG), or the combination of exercise and protein supplementation group. A significant interaction effect between body weight and body mass index (BMI) was observed, with the PrG (-2.1% body weight, -1.8% BMI) showing the highest reductions. Further, there was a decrease in % body fat (-4.5%) and an increase in fat-free mass (kg) (+5.7%) in the ExG after 12 weeks (P exercise program was found to improve muscle strength and body composition, while protein supplementation reduced body weight and increased upper body strength, among sarcopenic elderly in Malaysia. PMID:24143082

  8. Impact of extraneous proteins on the gastrointestinal fate of sunflower seed (Helianthus annuus) oil bodies: a simulated gastrointestinal tract study.

    Science.gov (United States)

    Makkhun, Sakunkhun; Khosla, Amit; Foster, Tim; McClements, David Julian; Grundy, Myriam M L; Gray, David A

    2015-01-01

    In this study, we examined the physicochemical nature of sunflower seed oil bodies (in the absence and presence of added protein) exposed to gastrointestinal conditions in vitro: crude oil bodies (COB); washed oil bodies (WOB); whey protein isolate-enriched oil bodies (WOB-WPI); and, sodium caseinate enriched-oil bodies (WOB-SC). All oil body emulsions were passed through an in vitro digestion model that mimicked the stomach and duodenal environments, and their physicochemical properties were measured before, during, and after digestion. Oil bodies had a positive charge under gastric conditions because the pH was below the isoelectric point of the adsorbed protein layer, but they had a negative charge under duodenal conditions which was attributed to changes in interfacial composition resulting from adsorption of bile salts. Oil bodies were highly susceptible to flocculation and coalescence in both gastric and duodenal conditions. SDS-PAGE analysis indicated degradation of oleosin proteins (ca. 18-21 kDa) to a greater or lesser extent (dependent on the emulsion) during the gastric phase in all emulsions tested; there is evidence that some oleosin remained intact in the crude oil body preparation during this phase of the digestion process. Measurements of protein displacement from the surface of COBs during direct exposure to bile salts, without inclusion of a gastric phase, indicated the removal of intact oleosin from native oil bodies. PMID:25284307

  9. Association of C - Reactive Protein and Body Mass Index with Duration of Mechanical Ventilation in

    OpenAIRE

    M. Safavi, M.D; A. Honarmand, M.D

    2007-01-01

    AbstractBackground and purpose: The aim of this study was to determine the incidence and presence of a relationship between predictors of body mass index (BMI) or C-reactive protein (CRP) and duration of mechanical ventilation, in trauma patients who were admitted to the intensive care unite (ICU). Furthermore, we compared their prognostic significance, with known indicators such as, the Sequential Organ Failure Assessment (SOFA) score.Materials and Methods: This prospective observational stu...

  10. Body composition and deposition efficiency of protein and energy in grazing young bulls

    Directory of Open Access Journals (Sweden)

    Eriton Egidio Lisboa Valente

    2014-05-01

    Full Text Available The effects of supplementation with different protein: carbohydrate ratios on body composition, carcass characteristics and protein and energy deposition efficiency of young were assessed. Twenty-four Nellorecalves (132.5 ± 5.5 kgand 90-150 days of age were kept on pasture for a 430 day experimental period. The treatments were: Control = mineral mixture only; HPHC = high-protein and high-carbohydrate supplement; HPLC = high-protein and low-carbohydrate supplement; LPHC = low-protein and high-carbohydrate supplement; LPLC = low-protein and low-carbohydrate supplement. Four animals at begning and 20 animal at end of experiment were slaughtered to evaluate the carcass composition. Control bulls had the lowest (p 0.05 between supplemented bulls (13 Mcal day-1. Although non-supplemented bulls had less (p 0.05 between supplemented bulls. High-carbohydrate supplements were associated with more (p 0.05 in the energy efficiency between the groups. Therefore, supplementation increases the intake and retention of protein and energy without changing the retention efficiency.

  11. Characterization of magnetized ore bodies based on three-component borehole magnetic and directional borehole seismic measurements

    Science.gov (United States)

    Virgil, Christopher; Neuhaus, Martin; Hördt, Andreas; Giese, Rüdiger; Krüger, Kay; Jurczyk, Andreas; Juhlin, Christopher; Juhojuntti, Niklas

    2016-04-01

    In the last decades magnetic prospecting using total field data was used with great success for localization and characterization of ferromagnetic ore bodies. Especially borehole magnetic measurements reveal important constraints on the extent and depth of potential mining targets. However, due to the inherent ambiguity of the interpretation of magnetic data, the resulting models of the distribution of magnetized material, such as iron ore bodies, are not entirely reliable. Variations in derived parameters like volume and estimated ore content of the expected body have significant impact on the economic efficiency of a planned mine. An important improvement is the introduction of three-component borehole magnetic sondes. Modern tools comprise orientation modules which allow the continuous determination of the tool's heading regardless of the well inclination and independent of the magnetic field. Using the heading information the recorded three-component magnetic data can be transferred from the internal tool's frame to the geographic reference frame. The vector information yields a more detailed and reliable description of the ore bodies compared to total field or horizontal and vertical field data. Nevertheless complementary information to constrain the model is still advisable. The most important supplementary information for the interpretation of magnetic data is the knowledge of the structural environment of the target regions. By discriminating dissimilar rock units, a geometrical starting model can be derived, constraining the magnetic interpretation and leading to a more robust estimation of the rock magnetizations distribution. The most common approach to reveal the lithological setting rests upon seismic measurements. However, for deep drilling targets surface seismic and VSP lack the required spatial resolution of 10s of meters. A better resolution is achieved by using directed sources and receivers inside the borehole. Here we present the application of

  12. The proteins of intra-nuclear bodies: a data-driven analysis of sequence, interaction and expression

    Directory of Open Access Journals (Sweden)

    Bodén Mikael

    2010-04-01

    Full Text Available Abstract Background Cajal bodies, nucleoli, PML nuclear bodies, and nuclear speckles are morpohologically distinct intra-nuclear structures that dynamically respond to cellular cues. Such nuclear bodies are hypothesized to play important regulatory roles, e.g. by sequestering and releasing transcription factors in a timely manner. While the nucleolus and nuclear speckles have received more attention experimentally, the PML nuclear body and the Cajal body are still incompletely characterized in terms of their roles and protein complement. Results By collating recent experimentally verified data, we find that almost 1000 proteins in the mouse nuclear proteome are known to associate with one or more of the nuclear bodies. Their gene ontology terms highlight their regulatory roles: splicing is confirmed to be a core activity of speckles and PML nuclear bodies house a range of proteins involved in DNA repair. We train support-vector machines to show that nuclear proteins contain discriminative sequence features that can be used to identify their intra-nuclear body associations. Prediction accuracy is highest for nucleoli and nuclear speckles. The trained models are also used to estimate the full protein complement of each nuclear body. Protein interactions are found primarily to link proteins in the nuclear speckles with proteins from other compartments. Cell cycle expression data provide support for increased activity in nucleoli, nuclear speckles and PML nuclear bodies especially during S and G2 phases. Conclusions The large-scale analysis of the mouse nuclear proteome sheds light on the functional organization of physically embodied intra-nuclear compartments. We observe partial support for the hypothesis that the physical organization of the nucleus mirrors functional modularity. However, we are unable to unambiguously identify proteins' intra-nuclear destination, suggesting that critical drivers behind of intra-nuclear translocation are yet to

  13. The human keratinocyte two-dimensional gel protein database (update 1995): mapping components of signal transduction pathways

    DEFF Research Database (Denmark)

    Celis, J E; Rasmussen, H H; Gromov, P;

    1995-01-01

    identified (protein name, organelle components, etc.) using a procedure or a combination of procedures that include (i) comigration with known human proteins, (ii) 2-D gel immunoblotting using specific antibodies, (iii) microsequencing of Coomassie Brilliant Blue stained proteins, (iv) mass spectrometry, (v...

  14. Direct interaction of mitochondrial targeting presequences with purified components of the TIM23 protein complex.

    Science.gov (United States)

    Marom, Milit; Dayan, Dana; Demishtein-Zohary, Keren; Mokranjac, Dejana; Neupert, Walter; Azem, Abdussalam

    2011-12-23

    Precursor proteins that are imported from the cytosol into the matrix of mitochondria carry positively charged amphipathic presequences and cross the inner membrane with the help of vital components of the TIM23 complex. It is currently unclear which subunits of the TIM23 complex recognize and directly bind to presequences. Here we analyzed the binding of presequence peptides to purified components of the TIM23 complex. The interaction of three different presequences with purified soluble domains of yeast Tim50 (Tim50IMS), Tim23 (Tim23IMS), and full-length Tim44 was examined. Using chemical cross-linking and surface plasmon resonance we demonstrate, for the first time, the ability of purified Tim50IMS and Tim44 to interact directly with the yeast Hsp60 presequence. We also analyzed their interaction with presequences derived from precursors of yeast mitochondrial 70-kDa heat shock protein (mHsp70) and of bovine cytochrome P450SCC. Moreover, we characterized the nature of the interactions and determined their KDs. On the basis of our results, we suggest a mechanism of translocation where stronger interactions of the presequences on the trans side of the channel support the import of precursor proteins through TIM23 into the matrix. PMID:21969381

  15. Direct Interaction of Mitochondrial Targeting Presequences with Purified Components of the TIM23 Protein Complex*

    Science.gov (United States)

    Marom, Milit; Dayan, Dana; Demishtein-Zohary, Keren; Mokranjac, Dejana; Neupert, Walter; Azem, Abdussalam

    2011-01-01

    Precursor proteins that are imported from the cytosol into the matrix of mitochondria carry positively charged amphipathic presequences and cross the inner membrane with the help of vital components of the TIM23 complex. It is currently unclear which subunits of the TIM23 complex recognize and directly bind to presequences. Here we analyzed the binding of presequence peptides to purified components of the TIM23 complex. The interaction of three different presequences with purified soluble domains of yeast Tim50 (Tim50IMS), Tim23 (Tim23IMS), and full-length Tim44 was examined. Using chemical cross-linking and surface plasmon resonance we demonstrate, for the first time, the ability of purified Tim50IMS and Tim44 to interact directly with the yeast Hsp60 presequence. We also analyzed their interaction with presequences derived from precursors of yeast mitochondrial 70-kDa heat shock protein (mHsp70) and of bovine cytochrome P450SCC. Moreover, we characterized the nature of the interactions and determined their KDs. On the basis of our results, we suggest a mechanism of translocation where stronger interactions of the presequences on the trans side of the channel support the import of precursor proteins through TIM23 into the matrix. PMID:21969381

  16. Post-translational modifications of chicken myelin basic protein charge components.

    Science.gov (United States)

    Kim, Jeongkwon; Zhang, Rui; Strittmatter, Eric F; Smith, Richard D; Zand, Robert

    2009-02-01

    Purified myelin basic protein (MBP) from various species contains several post-translationally modified forms termed charge components or charge isomers. Chicken MBP contains four charge components denoted as C1, C2, C3 and C8. (The C8 isomer is a complex mixture and was not investigated in this study.) These findings are in contrast to those found for human, bovine and other mammalian MBP's. Mammalian MBP's, each of which contain seven or eight charge components depending on the analysis of the CM-52 chromatographic curves and the PAGE gels obtained under basic pH conditions. Chicken MBP components C1, C2 and C3 were treated with trypsin and endoproteinase Glu-C. The resulting digests were analyzed by capillary liquid chromatography combined with either an ion trap tandem mass spectrometer or with a Fourier transform ion cyclotron resonance mass spectrometer. This instrumentation permitted establishing the amino acid composition and the determination of the post-translational modifications for each of the three charge components C1-C3. With the exception of N-terminal acetylation, the post-translational modifications were partial. The C1 component lacks any phosphorylated sites, a finding in agreement with the analysis of other MBP species. It also had a single methylation at R105 as did the components C2 and C3. The C2 component contains ten phosphorylated sites (S7, S18, S33, S64, S73, T96, S113, S141, S164, and S168), and modified arginine to citrulline residues at R24, and R165. Component C3 contains eight phosphorylated sites (S7, S33, S64, T96, S113, S141, S164, and S168), and citrulline residues at Arginine 41, R24 and R165. Partial deamidation of glutamine residues Q71, Q101 and Q146 were present in addition to asparagine N90 that was found in all three charge components. The glutamine at residue 3 is partially deamidated in isomers C1 and C2, whereas glutamine 74 and asparagine 83 were found not to be deamidated. Comparison of the PTM's of MBP's isolated

  17. Infection with the oncogenic human papillomavirus type 59 alters protein components of the cornified cell envelope

    International Nuclear Information System (INIS)

    Infection of the genital tract with human papillomaviruses (HPVs) leads to proliferative and dysplastic epithelial lesions. The mechanisms used by the virus to escape the infected keratinocyte are not well understood. Infection of keratinocytes with HPV does not cause lysis, the mechanism used by many viruses to release newly formed virions. For HPV 11, a type associated with a low risk of neoplastic disease, the cornified cell envelope (CCE) of infected keratinocytes is thin and fragile, and transcription of loricrin, the major CCE protein, is reduced. The effects of high-risk HPV infection on components of the CCE have not been previously reported. HPV 59, an oncogenic genital type related to HPV types 18 and 45 was identified in a condylomata acuminata lesion. An extract of this lesion was used to infect human foreskin fragments, which were grown in athymic mice as xenografts. Continued propagation using extracts of xenografts permitted growth of additional HPV 59-infected xenografts. CCEs purified from HPV 59-infected xenografts displayed subtle morphologic abnormalities compared to those derived from uninfected xenografts. HPV 59-infected xenografts revealed dysplastic-appearing cells with mitotic figures. Detection of loricrin, involucrin, and cytokeratin 10 was reduced in HPV 59-infected epithelium, while small proline-rich protein 3 (SPR3) was increased. Reduction in loricrin was most apparent in regions of epithelium containing abundant HPV 59 DNA. Compared to uninfected epithelium, loricrin transcription was decreased in HPV 59-infected epithelium. We conclude that HPV 59 shares with HPV 11 the ability to alter CCE components and to specifically reduce transcription of the loricrin gene. Because loricrin is the major CCE protein, a reduction in this component could alter the physical properties of the CCE, thus facilitating virion release

  18. Protein content and electrophoretic profile of fat body and ovary extracts from workers of Melipona quadrifasciata anthidioides (Hymenoptera, Meliponini

    Directory of Open Access Journals (Sweden)

    Vagner T. Paes de Oliveira

    Full Text Available Workers of Melipona quadrifasciata anthidioides (Lepeletier, 1836 develop their ovaries and lay eggs, therefore the production of vitellogenin is expected. In electrophoretic profiles only fat body extracts from nurse workers and ovary extracts from newly-emerged workers show protein with molecular mass similar to vitellogenin. However, an increase in the protein content was detected in forager fat body. This increase was attributed to storage of vitellogenin or other proteins in the previous phase and not discharged into the hemolymph or to an effect of the increased titre of juvenile hormone in this phase of worker life over the fat body functioning.

  19. Abnormal accumulation of prion protein mRNA in muscle fibers of patients with sporadic inclusion-body myositis and hereditary inclusion-body myopathy.

    OpenAIRE

    Sarkozi, E; Askanas, V.; Engel, W K

    1994-01-01

    Sporadic inclusion-body myositis is the most common progressive muscle disease of older patients. The muscle biopsy demonstrates mononuclear cell inflammation and vacuolated muscle fibers containing paired helical filaments and 6 to 10-nm fibrils, both resembling those of Alzheimer brain, and Congo-red positivity. Hereditary inclusion-body myopathy designates patients cytopathologically similar but without inflammation. In both muscle diseases, prion, and several proteins characteristic of Al...

  20. 'Sheltered disruption' of Neurospora crassa MOM22, an essential component of the mitochondrial protein import complex.

    OpenAIRE

    Nargang, F E; Künkele, K P; Mayer, A.; Ritzel, R G; Neupert, W; Lill, R

    1995-01-01

    MOM22 is a component of the protein import complex of the mitochondrial outer membrane of Neurospora crassa. Using the newly developed procedure of 'sheltered disruption', we created a heterokaryotic strain harboring two nuclei, one with a null allele of the mom-22 gene and the other with a wild-type allele. Homokaryons bearing the mom-22 disruption could not be isolated, suggesting that mom-22 is an essential gene. The mutant nucleus can be forced to predominate in the heterokaryon through t...

  1. Effects of protein intake and gender on body composition changes: a randomized clinical weight loss trial

    Directory of Open Access Journals (Sweden)

    Evans Ellen M

    2012-06-01

    Full Text Available Abstract Limited data on sex differences in body composition changes in response to higher protein diets (PRO compared to higher carbohydrate diets (CARB suggest that a PRO diet helps preserve lean mass (LM in women more so than in men. Objective To compare male and female body composition responses to weight loss diets differing in macronutrient content. Design Twelve month randomized clinical trial with 4mo of weight loss and 8mo weight maintenance. Subjects Overweight (N = 130; 58 male (M, 72 female (F; BMI = 32.5 ± 0.5 kg/m2 middle-aged subjects were randomized to energy-restricted (deficit ~500 kcal/d diets providing protein at 1.6 g.kg-1.d-1 (PRO or 0.8 g.kg-1.d-1 (CARB. LM and fat mass (FM were measured using dual X-ray absorptiometry. Body composition outcomes were tested in a repeated measures ANOVA controlling for sex, diet, time and their two- and three-way interactions at 0, 4, 8 and 12mo. Results When expressed as percent change from baseline, males and females lost similar amounts of weight at 12mo (M:-11.2 ± 7.1 %, F:-9.9 ± 6.0 %, as did diet groups (PRO:-10.7 ± 6.8 %, CARB:-10.1 ± 6.2 %, with no interaction of gender and diet. A similar pattern emerged for fat mass and lean mass, however percent body fat was significantly influenced by both gender (M:-18.0 ± 12.8 %, F:-7.3 ± 8.1 %, p  Conclusion PRO was more effective in reducing percent body fat vs. CARB over 12mo weight loss and maintenance. Men lost percent total body fat and trunk fat more effectively than women. No interactive effects of protein intake and gender are evident.

  2. Protein and carbohydrate components in the Risoe high-lysine barley mutants

    International Nuclear Information System (INIS)

    Seeds of the Risoe high-lysine barley mutant were analysed for nitrogenous and carbohydrate components to identify possible interactions between the high-lysine character and the low grain yield. The protein yields of the high-lysine mutants are 73-97% of that of the parents. In the parents protein is fractionated into approximately 40% prolamin (approximately 1% lysine), 55% non-prolamin protein (NPP) (6% lysine) and 5% non-protein nitrogen (NPN). In all high-lysine mutants NPP is increased and prolamin reduced. Neither different high-lysine genes nor large differences in N content in the seeds have any significant influence on the amino acid composition of NPP. The amounts of the main prolamin components, hordein-1 and hordein-2, are reduced in different and often characteristic ways in most of the mutants. The starch yields of the mutants are 38-88% of the yields of the parents. This decrease is mainly caused by a reduced content of starch per seed, but also by a reduced seed number. The latter may be independent of the high-lysine genes. There is no correlation between the amounts of starch and prolamin in the mutants, but both may be changed in the same direction in allelic mutants with different genetic background. The percentage of starch and soluble sugars in all the high-lysine mutants shows a highly significant negative correlation. This suggests a possible common mechanism responsible for the reduction in starch synthesis in all the high-lysine mutants. (author)

  3. Protein intake at 9 mo of age is associated with body size but not with body fat in 10-y-old Danish children

    DEFF Research Database (Denmark)

    Hoppe, Camilla; Mølgaard, Christian; Thomsen, Birthe Lykke Riegels;

    2004-01-01

    During the complementary feeding period, infants shift from a daily protein intake (PI) of approximately 1 g/kg body wt to an intake 3-4 times as high. A high PI probably has both endocrine and physiologic effects and may increase the risk of obesity.......During the complementary feeding period, infants shift from a daily protein intake (PI) of approximately 1 g/kg body wt to an intake 3-4 times as high. A high PI probably has both endocrine and physiologic effects and may increase the risk of obesity....

  4. Matrix gla protein (MGP): an overexpressed and migration-promoting mesenchymal component in glioblastoma

    International Nuclear Information System (INIS)

    Recent studies have demonstrated that a molecular subtype of glioblastoma is characterized by overexpression of extracellular matrix (ECM)/mesenchymal components and shorter survival. Specifically, gene expression profiling studies revealed that matrix gla protein (MGP), whose function has traditionally been linked to inhibition of calcification of arteries and cartilage, is overexpressed in glioblastomas and associated with worse outcome. In order to analyze the role of MGP in glioblastomas, we performed expression, migration and proliferation studies. Real-time PCR and ELISA assays confirmed overexpression of MGP in glioblastoma biopsy specimens and cell lines at mRNA and protein levels as compared to normal brain tissue. Immunohistochemistry verified positivity of glial tumor cells for MGP. RNAi-mediated knockdown of MGP in three glioma cell lines (U343MG, U373MG, H4) led to marked reduction of migration, as demonstrated by wound healing and transwell assays, while no effect on proliferation was seen. Our data suggest that upregulation of MGP (and possibly other ECM-related components as well) results in unfavorable prognosis via increased migration

  5. Smed-SmB, a member of the LSm protein superfamily, is essential for chromatoid body organization and planarian stem cell proliferation.

    Science.gov (United States)

    Fernandéz-Taboada, Enrique; Moritz, Sören; Zeuschner, Dagmar; Stehling, Martin; Schöler, Hans R; Saló, Emili; Gentile, Luca

    2010-04-01

    Planarians are an ideal model system to study in vivo the dynamics of adult pluripotent stem cells. However, our knowledge of the factors necessary for regulating the 'stemness' of the neoblasts, the adult stem cells of planarians, is sparse. Here, we report on the characterization of the first planarian member of the LSm protein superfamily, Smed-SmB, which is expressed in stem cells and neurons in Schmidtea mediterranea. LSm proteins are highly conserved key players of the splicing machinery. Our study shows that Smed-SmB protein, which is localized in the nucleus and the chromatoid body of stem cells, is required to safeguard the proliferative ability of the neoblasts. The chromatoid body, a cytoplasmatic ribonucleoprotein complex, is an essential regulator of the RNA metabolism required for the maintenance of metazoan germ cells. However, planarian neoblasts and neurons also rely on its functions. Remarkably, Smed-SmB dsRNA-mediated knockdown results in a rapid loss of organization of the chromatoid body, an impairment of the ability to post-transcriptionally process the transcripts of Smed-CycB, and a severe proliferative failure of the neoblasts. This chain of events leads to a quick depletion of the neoblast pool, resulting in a lethal phenotype for both regenerating and intact animals. In summary, our results suggest that Smed-SmB is an essential component of the chromatoid body, crucial to ensure a proper RNA metabolism and essential for stem cell proliferation. PMID:20215344

  6. High whey protein intake delayed the loss of lean body mass in healthy old rats, whereas protein type and polyphenol/antioxidant supplementation had no effects.

    Directory of Open Access Journals (Sweden)

    Laurent Mosoni

    Full Text Available Our aim was to compare and combine 3 nutritional strategies to slow down the age-related loss of muscle mass in healthy old rats: 1 increase protein intake, which is likely to stimulate muscle protein anabolism; 2 use leucine rich, rapidly digested whey proteins as protein source (whey proteins are recognized as the most effective proteins to stimulate muscle protein anabolism. 3 Supplement animals with a mixture of chamomile extract, vitamin E, vitamin D (reducing inflammation and oxidative stress is also effective to improve muscle anabolism. Such comparisons and combinations were never tested before. Nutritional groups were: casein 12% protein, whey 12% protein, whey 18% protein and each of these groups were supplemented or not with polyphenols/antioxidants. During 6 months, we followed changes of weight, food intake, inflammation (plasma fibrinogen and alpha-2-macroglobulin and body composition (DXA. After 6 months, we measured muscle mass, in vivo and ex-vivo fed and post-absorptive muscle protein synthesis, ex-vivo muscle proteolysis, and oxidative stress parameters (liver and muscle glutathione, SOD and total antioxidant activities, muscle carbonyls and TBARS. We showed that although micronutrient supplementation reduced inflammation and oxidative stress, the only factor that significantly reduced the loss of lean body mass was the increase in whey protein intake, with no detectable effect on muscle protein synthesis, and a tendency to reduce muscle proteolysis. We conclude that in healthy rats, increasing protein intake is an effective way to delay sarcopenia.

  7. High whey protein intake delayed the loss of lean body mass in healthy old rats, whereas protein type and polyphenol/antioxidant supplementation had no effects.

    Science.gov (United States)

    Mosoni, Laurent; Gatineau, Eva; Gatellier, Philippe; Migné, Carole; Savary-Auzeloux, Isabelle; Rémond, Didier; Rocher, Emilie; Dardevet, Dominique

    2014-01-01

    Our aim was to compare and combine 3 nutritional strategies to slow down the age-related loss of muscle mass in healthy old rats: 1) increase protein intake, which is likely to stimulate muscle protein anabolism; 2) use leucine rich, rapidly digested whey proteins as protein source (whey proteins are recognized as the most effective proteins to stimulate muscle protein anabolism). 3) Supplement animals with a mixture of chamomile extract, vitamin E, vitamin D (reducing inflammation and oxidative stress is also effective to improve muscle anabolism). Such comparisons and combinations were never tested before. Nutritional groups were: casein 12% protein, whey 12% protein, whey 18% protein and each of these groups were supplemented or not with polyphenols/antioxidants. During 6 months, we followed changes of weight, food intake, inflammation (plasma fibrinogen and alpha-2-macroglobulin) and body composition (DXA). After 6 months, we measured muscle mass, in vivo and ex-vivo fed and post-absorptive muscle protein synthesis, ex-vivo muscle proteolysis, and oxidative stress parameters (liver and muscle glutathione, SOD and total antioxidant activities, muscle carbonyls and TBARS). We showed that although micronutrient supplementation reduced inflammation and oxidative stress, the only factor that significantly reduced the loss of lean body mass was the increase in whey protein intake, with no detectable effect on muscle protein synthesis, and a tendency to reduce muscle proteolysis. We conclude that in healthy rats, increasing protein intake is an effective way to delay sarcopenia. PMID:25268515

  8. Location of Body Wave Microseism Sources Using Three-Component Data From a Large Aperture Seismic Array in China

    Science.gov (United States)

    Liu, Q.; Koper, K. D.; Burlacu, R.; Ni, S.; Wang, F.

    2015-12-01

    From September 2013 through October 2014 up to 100 Guralp CMG-3 broadband seismometers were deployed in the WT-Array (WTA) in northwest China. The aperture of WTA is about 700 km, with an average station spacing of approximately 50 km. Here, we process continuous, three-component WTA data to detect and locate body wave microseism sources in four distinct period bands: 1.0-2.5 s, 2.5-5 s, 5-10 s, and 10-20 s. We back-project vertical component data through a 1D reference Earth model (AK135) to a global grid of hypothetical source locations, assuming P-wave (30o-90o), PP-wave (60o-180o), and S-wave (30o-75o) propagation. At the same time, we rotate the horizontals and back-project the radial and transverse components of the wavefield. For each frequency band, grid point, and assumed origin time, the array power is calculated from the amplitude of a windowed, filtered, and tapered time domain beam constructed with fourth-root stacking. We find strong P-wave and S-wave noise sources in the North Pacific and North Atlantic Oceans. Shorter period sources (2.5-5 s) are mainly observed in the North Pacific Ocean, while both short and long period (2.5-20 s) sources are observed in the North Atlantic Ocean. Median power plots for each month during September 2013 through October 2014 show distinct seasonal variations. The energy peaks in the North Atlantic are visible from November to March and strong energy is also observed in the North Pacific from October to April. We also observe PP-waves in the Southern Ocean, especially for May-August 2014. Using classical f-k analysis and plane-wave propagation, we are able to confirm the back-projection results. To improve our understanding of body wave microseism generation, we compare the observed P, S, and PP wave microseism locations with the predictions of significant wave height and wave-wave interactions derived from the WAVEWATCH III ocean model.From September 2013 through October 2014 up to 100 Guralp CMG-3 broadband

  9. The Leishmania donovani lipophosphoglycan T lymphocyte-reactive component is a tightly associated protein complex.

    Science.gov (United States)

    Jardim, A; Tolson, D L; Turco, S J; Pearson, T W; Olafson, R W

    1991-11-15

    Lymphocytes from mice immunized with Leishmania donovani (LPG) were specifically stimulated to proliferate in vitro by purified LPG or its delipidated congener, phosphoglycan. The response was dose dependent and required prior immunization with either LPG or phosphoglycan. Proliferation was eliminated by specific depletion of Thy-1+ cells with antisera and C and the proliferating T cell subset was shown to be CD4+CD8-. Tests of various LPG fragments indicated that the T cell stimulation was associated with the core structure of LPG rather than the lipid or phosphoglycan repeat structure. However, amino acid analysis of LPG and active LPG fragments, after acid hydrolysis, showed the presence of amino acids in peptide linkage. Specific hydrolysis of the glycosidic linkages in LPG with trifluoromethanesulfonic acid provided polypeptide material reactive with two mAb previously believed to be LPG carbohydrate core specific. The protein was separated from LPG by reverse phase chromatography and shown to be a complex of proteins with common epitopes recognized by the two mAb. The dominant species isolated from LPG was a set of small, approximately 11,000 Mr, molecules. Subsequent T cell proliferation studies showed that the lymphocyte stimulation was associated with the protein component of LPG and not the glycan. PMID:1940354

  10. Effects of inulin on the structure and emulsifying properties of protein components in dough.

    Science.gov (United States)

    Liu, Juan; Luo, Denglin; Li, Xuan; Xu, Baocheng; Zhang, Xiaoyu; Liu, Jianxue

    2016-11-01

    High-purity gliadin, glutenin and gluten fractions were extracted from wheat gluten flour. To investigate the effects of three types of inulin with different degrees of polymerization (DP) on the emulsifying properties, disulfide contents, secondary structures and microstructures of these fractions, Turbidimetry, spectrophotometer, Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) were used in this study. The results showed that the emulsifying activity of gliadin was higher than that of glutenin and gluten, but its emulsion stability was lower than that of glutenin. Adding inulin increased the emulsifying activity of the three protein fractions and emulsion stability of gliadin and gluten, but decreased the emulsion stability of glutenin and disulfide bond contents of glutenin and gluten. In the presence of inulin, the α-helical structure of the three proteins had no significant change, whereas the β-turn structure decreased and β-sheet structure increased. The SEM images showed that inulin had the most significant effect on the glutenin microstructure. In general, inulin with a higher DP had greater effects on the structure and emulsifying properties of protein components in dough. PMID:27211643

  11. DELLA proteins are common components of symbiotic rhizobial and mycorrhizal signalling pathways.

    Science.gov (United States)

    Jin, Yue; Liu, Huan; Luo, Dexian; Yu, Nan; Dong, Wentao; Wang, Chao; Zhang, Xiaowei; Dai, Huiling; Yang, Jun; Wang, Ertao

    2016-01-01

    Legumes form symbiotic associations with either nitrogen-fixing bacteria or arbuscular mycorrhizal fungi. Formation of these two symbioses is regulated by a common set of signalling components that act downstream of recognition of rhizobia or mycorrhizae by host plants. Central to these pathways is the calcium and calmodulin-dependent protein kinase (CCaMK)-IPD3 complex which initiates nodule organogenesis following calcium oscillations in the host nucleus. However, downstream signalling events are not fully understood. Here we show that Medicago truncatula DELLA proteins, which are the central regulators of gibberellic acid signalling, positively regulate rhizobial symbiosis. Rhizobia colonization is impaired in della mutants and we provide evidence that DELLAs can promote CCaMK-IPD3 complex formation and increase the phosphorylation state of IPD3. DELLAs can also interact with NSP2-NSP1 and enhance the expression of Nod-factor-inducible genes in protoplasts. We show that DELLA is able to bridge a protein complex containing IPD3 and NSP2. Our results suggest a transcriptional framework for regulation of root nodule symbiosis. PMID:27514472

  12. On docking, scoring and assessing protein-DNA complexes in a rigid-body framework.

    Directory of Open Access Journals (Sweden)

    Marc Parisien

    Full Text Available We consider the identification of interacting protein-nucleic acid partners using the rigid body docking method FTdock, which is systematic and exhaustive in the exploration of docking conformations. The accuracy of rigid body docking methods is tested using known protein-DNA complexes for which the docked and undocked structures are both available. Additional tests with large decoy sets probe the efficacy of two published statistically derived scoring functions that contain a huge number of parameters. In contrast, we demonstrate that state-of-the-art machine learning techniques can enormously reduce the number of parameters required, thereby identifying the relevant docking features using a miniscule fraction of the number of parameters in the prior works. The present machine learning study considers a 300 dimensional vector (dependent on only 15 parameters, termed the Chemical Context Profile (CCP, where each dimension reflects a specific type of protein amino acid-nucleic acid base interaction. The CCP is designed to capture the chemical complementarities of the interface and is well suited for machine learning techniques. Our objective function is the Chemical Context Discrepancy (CCD, which is defined as the angle between the native system's CCP vector and the decoy's vector and which serves as a substitute for the more commonly used root mean squared deviation (RMSD. We demonstrate that the CCP provides a useful scoring function when certain dimensions are properly weighted. Finally, we explore how the amino acids on a protein's surface can help guide DNA binding, first through long-range interactions, followed by direct contacts, according to specific preferences for either the major or minor grooves of the DNA.

  13. Chromatoid Body Protein TDRD6 Supports Long 3’ UTR Triggered Nonsense Mediated mRNA Decay

    Science.gov (United States)

    Fanourgakis, Grigorios; Akpinar, Müge; Dahl, Andreas; Jessberger, Rolf

    2016-01-01

    Chromatoid bodies (CBs) are spermiogenesis-specific organelles of largely unknown function. CBs harbor various RNA species, RNA-associated proteins and proteins of the tudor domain family like TDRD6, which is required for a proper CB architecture. Proteome analysis of purified CBs revealed components of the nonsense-mediated mRNA decay (NMD) machinery including UPF1. TDRD6 is essential for UPF1 localization to CBs, for UPF1-UPF2 and UPF1-MVH interactions. Upon removal of TDRD6, the association of several mRNAs with UPF1 and UPF2 is disturbed, and the long 3’ UTR-stimulated but not the downstream exon-exon junction triggered pathway of NMD is impaired. Reduced association of the long 3’ UTR mRNAs with UPF1 and UPF2 correlates with increased stability and enhanced translational activity. Thus, we identified TDRD6 within CBs as required for mRNA degradation, specifically the extended 3’ UTR-triggered NMD pathway, and provide evidence for the requirement of NMD in spermiogenesis. This function depends on TDRD6-promoted assembly of mRNA and decay enzymes in CBs. PMID:27149095

  14. Alternative preparation of inclusion bodies excludes interfering non-protein contaminants and improves the yield of recombinant proinsulin

    OpenAIRE

    Mackin, Robert B.

    2014-01-01

    The goal of simple, high-yield expression and purification of recombinant human proinsulin has proven to be a considerable challenge. First, proinsulin forms inclusion bodies during bacterial expression. While this phenomenon can be exploited as a capture step, conventionally prepared inclusion bodies contain significant amounts of non-protein contaminants that interfere with subsequent chromatographic purification. Second, the proinsulin molecules within the inclusion bodies are incorrectly ...

  15. A Candidate Approach Implicates the Secreted Salmonella Effector Protein SpvB in P-Body Disassembly

    OpenAIRE

    Eulalio, A.; Fröhlich, K.; Mano, M; Giacca, M.; Vogel, J.

    2011-01-01

    P-bodies are dynamic aggregates of RNA and proteins involved in several post-transcriptional regulation processes. P-bodies have been shown to play important roles in regulating viral infection, whereas their interplay with bacterial pathogens, specifically intracellular bacteria that extensively manipulate host cell pathways, remains unknown. Here, we report that Salmonella infection induces P-body disassembly in a cell type-specific manner, and independently of previously characterized path...

  16. Modeling the time evolution of the nanoparticle-protein corona in a body fluid.

    Directory of Open Access Journals (Sweden)

    Daniele Dell'Orco

    Full Text Available BACKGROUND: Nanoparticles in contact with biological fluids interact with proteins and other biomolecules, thus forming a dynamic corona whose composition varies over time due to continuous protein association and dissociation events. Eventually equilibrium is reached, at which point the continued exchange will not affect the composition of the corona. RESULTS: We developed a simple and effective dynamic model of the nanoparticle protein corona in a body fluid, namely human plasma. The model predicts the time evolution and equilibrium composition of the corona based on affinities, stoichiometries and rate constants. An application to the interaction of human serum albumin, high density lipoprotein (HDL and fibrinogen with 70 nm N-iso-propylacrylamide/N-tert-butylacrylamide copolymer nanoparticles is presented, including novel experimental data for HDL. CONCLUSIONS: The simple model presented here can easily be modified to mimic the interaction of the nanoparticle protein corona with a novel biological fluid or compartment once new data will be available, thus opening novel applications in nanotoxicity and nanomedicine.

  17. Chronic Protein Restriction in Mice Impacts Placental Function and Maternal Body Weight before Fetal Growth.

    Directory of Open Access Journals (Sweden)

    Paula N Gonzalez

    Full Text Available Mechanisms of resource allocation are essential for maternal and fetal survival, particularly when the availability of nutrients is limited. We investigated the responses of feto-placental development to maternal chronic protein malnutrition to test the hypothesis that maternal low protein diet produces differential growth restriction of placental and fetal tissues, and adaptive changes in the placenta that may mitigate impacts on fetal growth. C57BL/6J female mice were fed either a low-protein diet (6% protein or control isocaloric diet (20% protein. On embryonic days E10.5, 17.5 and 18.5 tissue samples were prepared for morphometric, histological and quantitative RT-PCR analyses, which included markers of trophoblast cell subtypes. Potential endocrine adaptations were assessed by the expression of Prolactin-related hormone genes. In the low protein group, placenta weight was significantly lower at E10.5, followed by reduction of maternal weight at E17.5, while the fetuses became significantly lighter no earlier than at E18.5. Fetal head at E18.5 in the low protein group, though smaller than controls, was larger than expected for body size. The relative size and shape of the cranial vault and the flexion of the cranial base was affected by E17.5 and more severely by E18.5. The junctional zone, a placenta layer rich in endocrine and energy storing glycogen cells, was smaller in low protein placentas as well as the expression of Pcdh12, a marker of glycogen trophoblast cells. Placental hormone gene Prl3a1 was altered in response to low protein diet: expression was elevated at E17.5 when fetuses were still growing normally, but dropped sharply by E18.5 in parallel with the slowing of fetal growth. This model suggests that nutrients are preferentially allocated to sustain fetal and brain growth and suggests the placenta as a nutrient sensor in early gestation with a role in mitigating impacts of poor maternal nutrition on fetal growth.

  18. Chronic Protein Restriction in Mice Impacts Placental Function and Maternal Body Weight before Fetal Growth

    Science.gov (United States)

    Barbeito-Andrés, Jimena; Klenin, Natasha; Cross, James C.; Hallgrímsson, Benedikt

    2016-01-01

    Mechanisms of resource allocation are essential for maternal and fetal survival, particularly when the availability of nutrients is limited. We investigated the responses of feto-placental development to maternal chronic protein malnutrition to test the hypothesis that maternal low protein diet produces differential growth restriction of placental and fetal tissues, and adaptive changes in the placenta that may mitigate impacts on fetal growth. C57BL/6J female mice were fed either a low-protein diet (6% protein) or control isocaloric diet (20% protein). On embryonic days E10.5, 17.5 and 18.5 tissue samples were prepared for morphometric, histological and quantitative RT-PCR analyses, which included markers of trophoblast cell subtypes. Potential endocrine adaptations were assessed by the expression of Prolactin-related hormone genes. In the low protein group, placenta weight was significantly lower at E10.5, followed by reduction of maternal weight at E17.5, while the fetuses became significantly lighter no earlier than at E18.5. Fetal head at E18.5 in the low protein group, though smaller than controls, was larger than expected for body size. The relative size and shape of the cranial vault and the flexion of the cranial base was affected by E17.5 and more severely by E18.5. The junctional zone, a placenta layer rich in endocrine and energy storing glycogen cells, was smaller in low protein placentas as well as the expression of Pcdh12, a marker of glycogen trophoblast cells. Placental hormone gene Prl3a1 was altered in response to low protein diet: expression was elevated at E17.5 when fetuses were still growing normally, but dropped sharply by E18.5 in parallel with the slowing of fetal growth. This model suggests that nutrients are preferentially allocated to sustain fetal and brain growth and suggests the placenta as a nutrient sensor in early gestation with a role in mitigating impacts of poor maternal nutrition on fetal growth. PMID:27018791

  19. Identification of misfolded proteins in body fluids for the diagnosis of prion diseases.

    Science.gov (United States)

    Properzi, Francesca; Pocchiari, Maurizio

    2013-01-01

    Transmissible spongiform encephalopathy (TSE) or prion diseases are fatal rare neurodegenerative disorders affecting man and animals and caused by a transmissible infectious agent. TSE diseases are characterized by spongiform brain lesions with neuronal loss and the abnormal deposition in the CNS, and to less extent in other tissues, of an insoluble and protease resistant form of the cellular prion protein (PrP(C)), named PrP(TSE). In man, TSE diseases affect usually people over 60 years of age with no evident disease-associated risk factors. In some cases, however, TSE diseases are unequivocally linked to infectious episodes related to the use of prion-contaminated medicines, medical devices, or meat products as in the variant Creutzfeldt-Jakob disease (CJD). Clinical signs occur months or years after infection, and during this silent period PrP(TSE), the only reliable marker of infection, is not easily measurable in blood or other accessible tissues or body fluids causing public health concerns. To overcome the limit of PrP(TSE) detection, several highly sensitive assays have been developed, but attempts to apply these techniques to blood of infected hosts have been unsuccessful or not yet validated. An update on the latest advances for the detection of misfolded prion protein in body fluids is provided. PMID:24027585

  20. Estimation of whole body protein synthesis from oxidation of infused [1-14C]leucine

    International Nuclear Information System (INIS)

    In rats infused with NaH14CO3, 14CO2 exhalation approached an asymptotic rate equal to 96.0 +/- 1.3 (SD) % of the infusion rate of 14C in a monoexponential manner with a half-time of 20 min. In rats infused with [1-14C]leucine, 14CO2 exhalation (/0.96) approached an asymptotic value, (1-F), of 16-25% of 14C infusion rate (mean 20.4%) in a monoexponential manner with a half-time of approximately 31 min. Whole body protein synthesis (S) was calculated from 1-F and urea N plus ammonia N excretion (C) as S = CF/(1-F). S was a uniform function of body weight in these rats, in six additional rats in which S was measured 6 h after single intravenous injection of [1-14C]leucine and also in previously reported rats given single injections. The relationship was S (mg.100 g-1.h-1) = 11-0.143 body wt (g) +/- 8.3. In six of these rats, S was also estimated from the plateau specific activity of plasma leucine or plasma 2-ketoisocaproate (KIC); the former estimate of S was significantly lower (by an average of 17%), but S was the same when specific activity of KIC was employed. These results support the validity of the expired 14CO2 technique for measuring S

  1. SSX2 is a novel DNA-binding protein that antagonizes polycomb group body formation and gene repression

    DEFF Research Database (Denmark)

    Gjerstorff, Morten Frier; Relster, Mette Marie; Greve, Katrine Buch Viden;

    2014-01-01

    Polycomb group (PcG) complexes regulate cellular identity through epigenetic programming of chromatin. Here, we show that SSX2, a germline-specific protein ectopically expressed in melanoma and other types of human cancers, is a chromatin-associated protein that antagonizes BMI1 and EZH2 PcG body...

  2. The proteosomal degradation of fusion proteins cannot be predicted from the proteosome susceptibility of their individual components

    OpenAIRE

    Ilyinskii, Petr O.; Meriin, Anatoli B.; Gabai, Vladimir L.; Usachev, Evgeny V.; Prilipov, Alexei G.; Thoidis, Galini; Shneider, Alexander M.

    2008-01-01

    It is assumed that the proteosome-processing characteristics of fusion constructs can be predicted from the sum of the proteosome sensitivity of their components. In the present study, we observed that a fusion construct consisting of proteosome-degradable proteins does not necessarily result in a proteosome-degradable chimera. Conversely, fusion of proteosome-resistant proteins may result in a proteosome-degradable composite. We previously demonstrated that conserved influenza proteins can b...

  3. The effects of 8 weeks of whey or rice protein supplementation on body composition and exercise performance

    OpenAIRE

    Joy, Jordan M; Lowery, Ryan P; Wilson, Jacob M.; Purpura, Martin; De Souza, Eduardo O; Wilson, Stephanie MC; Kalman, Douglas S; Dudeck, Joshua E; Jäger, Ralf

    2013-01-01

    Background Consumption of moderate amounts of animal-derived protein has been shown to differently influence skeletal muscle hypertrophy during resistance training when compared with nitrogenous and isoenergetic amounts of plant-based protein administered in small to moderate doses. Therefore, the purpose of the study was to determine if the post-exercise consumption of rice protein isolate could increase recovery and elicit adequate changes in body composition compared to equally dosed whey ...

  4. TTP and BRF proteins nucleate processing body formation to silence mRNAs with AU-rich elements

    OpenAIRE

    Franks, Tobias M.; Lykke-Andersen, Jens

    2007-01-01

    In mammalian cells, mRNAs with AU-rich elements (AREs) are targeted for translational silencing and rapid degradation. Here we present evidence that in human cells the proteins Tristetraprolin (TTP) and BRF-1 deliver ARE-mRNAs to processing bodies (PBs), cytoplasmic assemblies of mRNAs, and associated factors that promote translational silencing and mRNA decay. First, depletion of endogenous TTP and BRF proteins, or overexpression of dominant-negative mutant TTP proteins, impairs the localiza...

  5. Oxidative stress-induced assembly of PML nuclear bodies controls sumoylation of partner proteins.

    Science.gov (United States)

    Sahin, Umut; Ferhi, Omar; Jeanne, Marion; Benhenda, Shirine; Berthier, Caroline; Jollivet, Florence; Niwa-Kawakita, Michiko; Faklaris, Orestis; Setterblad, Niclas; de Thé, Hugues; Lallemand-Breitenbach, Valérie

    2014-03-17

    The promyelocytic leukemia (PML) protein organizes PML nuclear bodies (NBs), which are stress-responsive domains where many partner proteins accumulate. Here, we clarify the basis for NB formation and identify stress-induced partner sumoylation as the primary NB function. NB nucleation does not rely primarily on intermolecular interactions between the PML SUMO-interacting motif (SIM) and SUMO, but instead results from oxidation-mediated PML multimerization. Oxidized PML spherical meshes recruit UBC9, which enhances PML sumoylation, allow partner recruitment through SIM interactions, and ultimately enhance partner sumoylation. Intermolecular SUMO-SIM interactions then enforce partner sequestration within the NB inner core. Accordingly, oxidative stress enhances NB formation and global sumoylation in vivo. Some NB-associated sumoylated partners also become polyubiquitinated by RNF4, precipitating their proteasomal degradation. As several partners are protein-modifying enzymes, NBs could act as sensors that facilitate and confer oxidative stress sensitivity not only to sumoylation but also to other post-translational modifications, thereby explaining alterations of stress response upon PML or NB loss. PMID:24637324

  6. The Components of Flemingia macrophylla Attenuate Amyloid β-Protein Accumulation by Regulating Amyloid β-Protein Metabolic Pathway

    Directory of Open Access Journals (Sweden)

    Yun-Lian Lin

    2012-01-01

    Full Text Available Flemingia macrophylla (Leguminosae is a popular traditional remedy used in Taiwan as anti-inflammatory, promoting blood circulation and antidiabetes agent. Recent study also suggested its neuroprotective activity against Alzheimer's disease. Therefore, the effects of F. macrophylla on Aβ production and degradation were studied. The effect of F. macrophylla on Aβ metabolism was detected using the cultured mouse neuroblastoma cells N2a transfected with human Swedish mutant APP (swAPP-N2a cells. The effects on Aβ degradation were evaluated on a cell-free system. An ELISA assay was applied to detect the level of Aβ1-40 and Aβ1-42. Western blots assay was employed to measure the levels of soluble amyloid precursor protein and insulin degrading enzyme (IDE. Three fractions of F. macrophylla modified Aβ accumulation by both inhibiting β-secretase and activating IDE. Three flavonoids modified Aβ accumulation by activating IDE. The activated IDE pool by the flavonoids was distinctly regulated by bacitracin (an IDE inhibitor. Furthermore, flavonoid 94-18-13 also modulates Aβ accumulation by enhancing IDE expression. In conclusion, the components of F. macrophylla possess the potential for developing new therapeutic drugs for Alzheimer's disease.

  7. Complement component 4

    Science.gov (United States)

    ... may have lower-than-normal levels of the complement proteins C3 and C4 . Complement activity varies throughout the body. ... Saunders; 2013:chap 6. Read More Cirrhosis Complement Complement component 3 (C3) Glomerulonephritis Hepatitis Hereditary angioedema Kidney transplant Lupus nephritis ...

  8. Effectiveness of exercise and protein supplementation intervention on body composition, functional fitness, and oxidative stress among elderly Malays with sarcopenia

    Directory of Open Access Journals (Sweden)

    Shahar S

    2013-10-01

    Full Text Available Suzana Shahar,1 Norshafarina Shari Kamaruddin,2 Manal Badrasawi,1 Noor Ibrahim Mohamed Sakian,3 Zahara Abd Manaf,1 Zaitun Yassin,4 Leonard Joseph51Dietetic Programme, 2Biomedical Programme, 3Occupational Therapy Programme, School of Healthcare Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, Kuala Lumpur, 4Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, 5Department of Physiotherapy, School of Healthcare Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, Kuala Lumpur, MalaysiaAbstract: Sarcopenia, characterized as muscle loss that occurs with aging, is a major health problem in an aging population, due to its implications on mobility, quality of life, and fall risk. Protein supplementation could improve the physical fitness by increasing protein anabolism, and exercise has a documented evidence of positive effect on functional status among the elderly. However, the combined effect of both protein supplementation and exercise has not been investigated among sarcopenic elderly in the Asian population. Thus, this study aimed to determine the effectiveness of exercise intervention and protein supplementation either alone or in combination for 12 weeks, on body composition, functional fitness, and oxidative stress among elderly Malays with sarcopenia. Sixty five sarcopenic elderly Malays aged 60-74 years were assigned to the control group, exercise group (ExG, protein supplementation group (PrG, or the combination of exercise and protein supplementation group. A significant interaction effect between body weight and body mass index (BMI was observed, with the PrG (-2.1% body weight, -1.8% BMI showing the highest reductions. Further, there was a decrease in % body fat (-4.5% and an increase in fat-free mass (kg (+5.7% in the ExG after 12 weeks (P < 0.05. The highest increments in lower and upper body strength were observed in the Pr

  9. Calcium Distribution in Globoid Crystals of Cucurbita Cotyledon Protein Bodies 1

    Science.gov (United States)

    Lott, John N. A.; Spitzer, Ernest; Vollmer, Catherine M.

    1979-01-01

    Energy-dispersive x-ray analysis was used to investigate the location of globoid crystals with relatively high Ca levels within cotyledons of Cucurbita maxima, Cucurbita mixta, and Cucurbita andreana. The small globoid crystals in both upper and lower epidermal cells commonly contained Ca. Ca was present in globoid crystals of all provascular regions with the exception of the very small provascular regions of C. maxima. In C. maxima and C. mixta cotyledons, some cases were observed where Ca was found in the globoid crystals of the first layer of mesophyll cells surrounding the provascular region, but in general Ca was absent from globoid crystals of palisade and spongy mesophyll cells. In C. andreana, globoid crystals of palisade and spongy mesophyll cells commonly contained at least some Ca. Cell position and cell type are factors affecting the Ca content of globoid crystals in protein bodies. PMID:16660825

  10. Interaction between genetic predisposition to adiposity and dietary protein in relation to subsequent change in body weight and waist circumference

    DEFF Research Database (Denmark)

    Ankarfeldt, Mikkel Z; Larsen, Sofus C; Ängquist, Lars;

    2014-01-01

    ) and dietary protein in relation to subsequent change in body weight (ΔBW) or change in WC (ΔWC). DESIGN: Three different Danish cohorts were used. In total 7,054 individuals constituted the study population with information on diet, 50 single-nucleotide polymorphisms (SNPs) associated with BMI, WC or......BACKGROUND: Genetic predisposition to adiposity may interact with dietary protein in relation to changes of anthropometry. OBJECTIVE: To investigate the interaction between genetic predisposition to higher body mass index (BMI), waist circumference (WC) or waist-hip ratio adjusted for BMI (WHRBMI......-score: <0.1 mm/y/5 energy% protein/risk allele, [-0.1; 0.1]). Similar results were seen when protein replaced fat. CONCLUSION: This study indicates that the genetic predisposition to general and abdominal adiposity, assessed by gene-scores, does not seem to modulate the influence of dietary protein on ΔBW...

  11. Monte Carlo study of thermal flux profiles and body correction factors for body protein measurements of obese subjects

    International Nuclear Information System (INIS)

    In previous calculations for total body nitrogen measurements of children, the anterior/posterior thermal neutron flux profile with depth was found to be fairly flat after an initial rise. However, for obese adults significant variations are found in the flux profile with the central flux value being as low as 20% of the peak value. The significance of these flux variations is examined. Correction factors are calculated for the varying attenuation of the nitrogen and hydrogen photons by a range of obese bodies. The calculations included the effect of the thermal flux profile as well as that of an outer layer of low nitrogen content adipose tissue. The bodies are assumed to have a homogeneous hydrogen content. A study of four obese body models with varying sex and fat content shows that the correction factors do not vary much between males and females. This is surprising since the female models are assumed to have a surface fat layer twice as thick as for the male models. The correction factors are found to be only slightly sensitive to the thermal flux variations with depth. 5 refs., 1 tab., 4 figs

  12. Soy versus whey protein bars: Effects on exercise training impact on lean body mass and antioxidant status

    Directory of Open Access Journals (Sweden)

    Babaknia Ari

    2004-12-01

    Full Text Available Abstract Background Although soy protein may have many health benefits derived from its associated antioxidants, many male exercisers avoid soy protein. This is due partly to a popular, but untested notion that in males, soy is inferior to whey in promoting muscle weight gain. This study provided a direct comparison between a soy product and a whey product. Methods Lean body mass gain was examined in males from a university weight training class given daily servings of micronutrient-fortified protein bars containing soy or whey protein (33 g protein/day, 9 weeks, n = 9 for each protein treatment group. Training used workouts with fairly low repetition numbers per set. A control group from the class (N = 9 did the training, but did not consume either type protein bar. Results Both the soy and whey treatment groups showed a gain in lean body mass, but the training-only group did not. The whey and training only groups, but not the soy group, showed a potentially deleterious post-training effect on two antioxidant-related related parameters. Conclusions Soy and whey protein bar products both promoted exercise training-induced lean body mass gain, but the soy had the added benefit of preserving two aspects of antioxidant function.

  13. Proteomic analysis of oil body membrane proteins accompanying the onset of desiccation phase during sunflower seed development.

    Science.gov (United States)

    Thakur, Anita; Bhatla, Satish C

    2015-01-01

    A noteworthy metabolic signature accompanying oil body (OB) biogenesis during oilseed development is associated with the modulation of the oil body membranes proteins. Present work focuses on 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE)-based analysis of the temporal changes in the OB membrane proteins analyzed by LC-MS/MS accompanying the onset of desiccation (20-30 d after anthesis; DAA) in the developing seeds of sunflower (Helianthus annuus L.). Protein spots unique to 20-30 DAA stages were picked up from 2-D gels for identification and the identified proteins were categorized into 7 functional classes. These include proteins involved in energy metabolism, reactive oxygen scavenging, proteolysis and protein turnover, signaling, oleosin and oil body biogenesis-associated proteins, desiccation and cytoskeleton. At 30 DAA stage, exclusive expressions of enzymes belonging to energy metabolism, desiccation and cytoskeleton were evident which indicated an increase in the metabolic and enzymatic activity in the cells at this stage of seed development (seed filling). Increased expression of cruciferina-like protein and dehydrin at 30 DAA stage marks the onset of desiccation. The data has been analyzed and discussed to highlight desiccation stage-associated metabolic events during oilseed development. PMID:26786011

  14. Polygalacturonase-inhibiting protein is a structural component of plant cell wall.

    Science.gov (United States)

    Protsenko, M A; Buza, N L; Krinitsyna, A A; Bulantseva, E A; Korableva, N P

    2008-10-01

    It is generally believed that plants "evolved a strategy of defending themselves from a phytopathogen attack" during evolution. This metaphor is used frequently, but it does not facilitate understanding of the mechanisms providing plant resistance to the invasion of foreign organisms and to other unfavorable external factors, as well as the role of these mechanisms in plant growth and development. Information on processes involving one of the plant resistance factors--polygalacturonase-inhibiting protein (PGIP)--is considered in this review. The data presented here indicate that PGIP, being an extracellular leucine-rich repeat-containing protein, performs important functions in the structure of plant cell wall. Amino acid residues participating in PGIP binding to homogalacturonan in the cell wall have been determined. The degree of methylation and the mode of distribution of homogalacturonan methyl groups are responsible for the formation of a complex structure, which perhaps determines the specificity of PGIP binding to pectin. PGIP is apparently one of the components of plant cell wall determining some of its mechanical properties; it is involved in biochemical processes related to growth, expansion, and maceration, and it influences plant morphology. Polygalacturonase (PG) is present within practically all plant tissues, but the manifestation of its activity varies significantly depending on physiological conditions in the tissue. Apparently, the regulation of PG functioning in apoplast significantly affects the development of processes associated with the modification of the structure of plant cell wall. PGIP can regulate PG activity through binding to homogalacturonan. The genetically determined structure of PGIP in plants determines the mode of its interaction with an invader and perhaps is one of the factors responsible for the set of pathogens causing diseases in a given plant species. PMID:18991551

  15. Generation of atomic four-body statistical potentials derived from the delaunay tessellation of protein structures.

    Science.gov (United States)

    Masso, Majid

    2012-01-01

    Delaunay tessellation of the atomic coordinates for a crystallographic protein structure yields an aggregate of non-overlapping and space-filling irregular tetrahedral simplices. The vertices of each simplex objectively identify a quadruplet of nearest neighbor atoms in the protein. Here we apply Delaunay tessellation to 1417 high-resolution structures of single chains that share low sequence identity, for the purpose of determining the relative frequencies of occurrence for all possible nearest neighbor atomic quadruplet types. Alternative distributions are explored by varying two fundamental parameters: atomic alphabet selection and cutoff length for admissible simplex edges. The distributions are then converted to four-body potential functions by implementing the inverted Boltzmann principle, which requires calculating the distribution of the reference state. Two alternative definitions for the reference state are presented, which introduces a third parameter, and we derive and compare an array of such potential functions. These knowledge-based statistical potentials based on higher-order interactions complement and generalize the more commonly encountered atom-pair potentials, for which a number of approaches are described in the literature. PMID:23367374

  16. Effects of protein and energy deficiency on the incorporation of 14C-Chlorella protein hydrolysate into body constituents of adult rats

    International Nuclear Information System (INIS)

    The effects of protein and/or energy deficiency on 14C incorporation into body constituents and 14C output in expired air and urine were investigated in adult rats using 14C-Chlorella protein hydrolysate. Rats were given a protein-free diet (PFD) for 2 weeks and conrol rats were fed ad libitum or pari-fed with the PFD group on a 12% lactalbumin diet (LA and Pair-fed, respectively). On the 15th day, animals received 14C-Chlorella protein hydolysate with 5 g of their respective diet. One group of PFD animals was given tracer by stomach tube without food (PFD-fast). Normal control rats ate about twice as much diet as the PFD group. The respiratory 14C output in the PFD group was identical with those in the LA and Pair-fed groups and was less than that in the PFD-fast group. The rate of protein synthesis, provisionally expressed as relative specific radioactivity, was more in the PFD group than in the normal group in the liver and less than the latter in the muscle. The LA group retained less total radioactivity in the body than the Pair-fed or PFD group, indicating high capability to hold the body protein in protein deficiency. In addition, decreased conversion of amino acids to lipids and glycogen was observed in the PFD group. All these differences are interpreted as adaptations to protein shortage. On prolonged fasting (PFD-fast group), gluconeogenesis in the liver increased to provide energy, despite the protein deficiency. The relative importances of protein and energy for tissue protein synthesis are briefly discussed. (author)

  17. Dissection of Molecular Mechanisms Regulating Protein Body Formation in Maize Endosperm - DE-FG03-95-ER20183

    International Nuclear Information System (INIS)

    Dissection of Molecular Mechanisms Regulating Protein Body Formation in Maize Endosperm - DE-FG03-95-ER20183 Final Technical Report and Patent Summary Dr. Brian A. Larkins, Department of Plant Sciences, University of Arizona, Tucson, AZ 85721 Endosperm texture is an important quality trait in maize, as it influences the shipping characteristics of the grain, its susceptibility to insects, the yield of grits from dry milling, energy costs during wet milling, and the baking and digestibility properties of the flour. There appears to be a causal relationship between kernel hardness and the formation of zein-containing protein bodies, as mutations affecting protein body number and structure are associated with a soft, starchy kernel. In this project we used a variety of approaches to better understand this relationship and investigate the molecular and biochemical changes associated with starchy endosperm mutants. We characterized the distribution of zein mRNAs on endosperm rough endoplasmic reticulum (RER) membranes and the interactions between zein proteins, as each of these could influence the structure of protein bodies. Based on in situ hybridization, mRNAs encoding the 22-kD alpha- and 27-kD gamma-zeins are randomly distributed on RER; hence, mRNA targeting does not appear to influence the formation of protein bodies. Investigation of the interactions between zein proteins (alpha, beta, gamma, delta) with the yeast two-hybrid system showed that interactions between the 19- and 22-alpha-zeins are relatively weak, although each of them interacted strongly with the 10-kD delta-zein. Strong interactions were detected between the alpha- and delta-zeins and the 16-kD gamma- and 15-kD beta-zeins; however, the 50-kD and 27-kD gamma-zeins did not interact detectably with the alpha- and delta-zein proteins. The NH2- and COOH-terminal domains of the 22-kD alpha-zein were found to interact most strongly with the 15-kD beta- and 16-kD gamma-zeins, suggesting the 16-kD and 15

  18. 植物油脂体及其结构蛋白研究%Study on Plant Oil Body and Its Structural Proteins

    Institute of Scientific and Technical Information of China (English)

    屠宝玉; 贾美慧; 李立新

    2011-01-01

    高等植物在其种子中储藏大量的脂类,为种子萌发及后续的籽苗生长提供碳源和能源.储藏脂类均贮存于油脂体(oil body,OB)中.油脂体的结构简单,核心部分是TAG 组成的不透明结构,外层为磷脂单分子层,表面有多种膜蛋白,其中含量最丰富的是 Oleosin.Oleosin 和Caleosin 作为2种结构蛋白在维持油脂体稳定性以及控制油脂体大小方面发挥重要的作用,对于种子质量的提高即种子油类产量和优化储存脂类及蛋白的提取过程具有重要的研究价值.油脂体表面还存在一些酶类如 Steroleosin,可能与甾类化合物的合成相关.对油脂体形成的机制及相关蛋白结构与功能的研究进展进行了概述.%Higher plants accumulate a large amount of storage lipids in their seeds, which provide carbon and energy sources for seed germination and subsequent seedling growth. Storage lipids are all stored in lipid body. The structure of oil body is simple, the central component are opaque structure what composed by TAG,and external surface are phospholipids single molecule layer. There is variety of membrane-associated proteins on the surface of oil body,and Oleosin is the majority. As structural proteins,Oleosin and Caleosin play important roles on maintaining stability and controlling size of oil body. They also have important research value on improving seed quality ,oil yield and optimization of extraction method of seed storage lipid and protein. There are also some enzymes on the surface ofoil body,such as Steroleosin,which may have something to do with steroid compound. This paper summarizes the mechanism of formation of oil body and recent research progress on related protein function.

  19. Body weight management effect of burdock (Arctium lappa L.) root is associated with the activation of AMP-activated protein kinase in human HepG2 cells.

    Science.gov (United States)

    Kuo, Daih-Huang; Hung, Ming-Chi; Hung, Chao-Ming; Liu, Li-Min; Chen, Fu-An; Shieh, Po-Chuen; Ho, Chi-Tang; Way, Tzong-Der

    2012-10-01

    Burdock (Arcticum lappa L.) root is used in folk medicine and also as a vegetable in Asian countries. In the present study, burdock root treatment significantly reduced body weight in rats. To evaluate the bioactive compounds, we successively extracted the burdock root with ethanol (AL-1), and fractionated it with n-hexane (AL-2), ethyl acetate (AL-3), n-butanol (AL-4), and water (AL-5). Among these fractions, AL-2 contained components with the most effective hypolipidemic potential in human hepatoma HepG2 cells. AL-2 decreased the expression of fatty acid synthase (FASN) and inhibited the activity of acetyl-coenzyme A carboxylase (ACC) by stimulating AMP-activated protein kinase (AMPK) through the LKB1 pathway. Three active compounds were identified from the AL-2, namely α-linolenic acid, methyl α-linolenate, and methyl oleate. These results suggest that burdock root is expected to be useful for body weight management. PMID:25005949

  20. Improving protein delivery of fibroblast growth factor-2 from bacterial inclusion bodies used as cell culture substrates

    OpenAIRE

    Seras Franzoso, Joaquin; Peebo, Karl; Garcia Fruitós, Elena; Vázquez Gómez, Esther; Rinas, Ursula; Villaverde Corrales, Antonio

    2014-01-01

    Altres ajuts: We are indebted CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN, Spain) for funding our research on inclusion bodies. Bacterial inclusion bodies (IBs) have recently been used to generate biocompatible cell culture interfaces, with diverse effects on cultured cells such as cell adhesion enhancement, stimulation of cell growth or induction of mesenchymal stem cell differentiation. Additionally, novel applications of IBs as sustained protein delivery systems with...

  1. Production and purification of avian antibodies (IgYs from inclusion bodies of a recombinant protein central in NAD+ metabolism

    Directory of Open Access Journals (Sweden)

    Paula A. Moreno-González

    2013-08-01

    Full Text Available The use of hens for the production of polyclonal antibodies reduces animal intervention and moreover yields a higher quantity of antibodies than other animal models.  The phylogenetic distance between bird and mammal antigens, often leads to more specific avian antibodies than their mammalian counterparts.Since a large amount of antigen is required for avian antibody production, the use of recombinant proteins for this procedure has been growing faster over the last years. Nevertheless, recombinant protein production through heterologous systems frequently prompts the protein to precipitate, forming insoluble aggregates of limited utility (inclusion bodies. A methodology for the production of avian polyclonal antibodies, using recombinant protein from inclusion bodies is presented in this article.In order to produce the antigen, a recombinant Nicotinamide mononucleotide adenylyltransferase from Giardia intestinalis (His-GiNMNAT was expressed in Escherichia coli.  The protein was purified through solubilization from inclusion bodies prior to its renaturalization.  Antibodies were purified from egg yolk of immunized hens by water dilution, followed by ammonium sulfate precipitation and thiophilic affinity chromatography.The purified antibodies were tested against His-GiNMNAT protein in Western blot essays. From one egg yolk, 14.4 mg of highly pure IgY were obtained; this antibody was able to detect 15ng of His-GiNMNAT.  IgY specificity was improved by means of antigen affinity purification, allowing its use for parasite protein recognition.

  2. A controlled trial of protein enrichment of meal replacements for weight reduction with retention of lean body mass

    Directory of Open Access Journals (Sweden)

    Bowerman Susan

    2008-08-01

    Full Text Available Abstract Background While high protein diets have been shown to improve satiety and retention of lean body mass (LBM, this study was designed to determine effects of a protein-enriched meal replacement (MR on weight loss and LBM retention by comparison to an isocaloric carbohydrate-enriched MR within customized diet plans utilizing MR to achieve high protein or standard protein intakes. Methods Single blind, placebo-controlled, randomized outpatient weight loss trial in 100 obese men and women comparing two isocaloric meal plans utilizing a standard MR to which was added supplementary protein or carbohydrate powder. MR was used twice daily (one meal, one snack. One additional meal was included in the meal plan designed to achieve individualized protein intakes of either 1 2.2 g protein/kg of LBM per day [high protein diet (HP] or 2 1.1 g protein/kg LBM/day standard protein diet (SP. LBM was determined using bioelectrical impedance analysis (BIA. Body weight, body composition, and lipid profiles were measured at baseline and 12 weeks. Results Eighty-five subjects completed the study. Both HP and SP MR were well tolerated, with no adverse effects. There were no differences in weight loss at 12 weeks (-4.19 ± 0.5 kg for HP group and -3.72 ± 0.7 kg for SP group, p > 0.1. Subjects in the HP group lost significantly more fat weight than the SP group (HP = -1.65 ± 0.63 kg; SP = -0.64 ± 0.79 kg, P = 0.05 as estimated by BIA. There were no significant differences in lipids nor fasting blood glucose between groups, but within the HP group a significant decrease in cholesterol and LDL cholesterol was noted at 12 weeks. This was not seen in the SP group. Conclusion Higher protein MR within a higher protein diet resulted in similar overall weight loss as the standard protein MR plan over 12 weeks. However, there was significantly more fat loss in the HP group but no significant difference in lean body mass. In this trial, subject compliance with both the

  3. Phosphoproteomic analysis of protein kinase C signaling in Saccharomyces cerevisiae reveals Slt2 mitogen-activated protein kinase (MAPK)-dependent phosphorylation of eisosome core components.

    Science.gov (United States)

    Mascaraque, Victoria; Hernáez, María Luisa; Jiménez-Sánchez, María; Hansen, Rasmus; Gil, Concha; Martín, Humberto; Cid, Víctor J; Molina, María

    2013-03-01

    The cell wall integrity (CWI) pathway of the model organism Saccharomyces cerevisiae has been thoroughly studied as a paradigm of the mitogen-activated protein kinase (MAPK) pathway. It consists of a classic MAPK module comprising the Bck1 MAPK kinase kinase, two redundant MAPK kinases (Mkk1 and Mkk2), and the Slt2 MAPK. This module is activated under a variety of stimuli related to cell wall homeostasis by Pkc1, the only member of the protein kinase C family in budding yeast. Quantitative phosphoproteomics based on stable isotope labeling of amino acids in cell culture is a powerful tool for globally studying protein phosphorylation. Here we report an analysis of the yeast phosphoproteome upon overexpression of a PKC1 hyperactive allele that specifically activates CWI MAPK signaling in the absence of external stimuli. We found 82 phosphopeptides originating from 43 proteins that showed enhanced phosphorylation in these conditions. The MAPK S/T-P target motif was significantly overrepresented in these phosphopeptides. Hyperphosphorylated proteins provide putative novel targets of the Pkc1-cell wall integrity pathway involved in diverse functions such as the control of gene expression, protein synthesis, cytoskeleton maintenance, DNA repair, and metabolism. Remarkably, five components of the plasma-membrane-associated protein complex known as eisosomes were found among the up-regulated proteins. We show here that Pkc1-induced phosphorylation of the eisosome core components Pil1 and Lsp1 was not exerted directly by Pkc1, but involved signaling through the Slt2 MAPK module. PMID:23221999

  4. A dynamic model to predict fat and protein fluxes and dry matter intake associated with body reserve changes in cattle.

    Science.gov (United States)

    Tedeschi, Luis O; Fox, Danny G; Kononoff, Paul J

    2013-04-01

    The objective of this paper was to develop the structure and concepts of a dynamic model to simulate dry matter intake (DMI) pattern and the fluxes of fat and protein in the body reserves of cattle associated with changes in body condition score (BCS) for application within the structure of applied nutrition models. This model was developed to add the capability of evaluating the effects of factors affecting pre- and postcalving DMI, daily energy and protein balances, and changes in BCS over a reproductive cycle. Input variables are average DMI, diet metabolizable energy, and animal information (body weight, BCS, milk production, and calf birth body weight) from each diet fed over the reproductive cycle. Because the depletion and repletion of body reserves in cattle is a complex system of coordinated metabolic processes that reflect hormonal and physiological changes caused by negative or positive energy balances, the system dynamics modeling methodology was used to develop this model. The model was used to evaluate the effect of the dynamic interactions between dietary supply and animal requirements for energy and protein on the fluxes of body fat and body protein of dairy cows over the reproductive cycle and Monte Carlo simulations were used to assess the sensitivity of the parameters. The main long-term factor affecting DMI pattern was the growth of the gravid uterus causing an increase in the volume of abdominal organs and a compression of the rumen, consequentially reducing feed intake. Changes in body reserves (fat and protein) were computed based on metabolizable energy balance, assuming different efficiency of utilization coefficients for fat and protein during repletion and mobilization. The model was evaluated with data from 37 dairy cows individually fed 3 different diets over the lactation and dry periods. The model was successful in simulating the observed pattern of DMI (mean square error was 3.59, 3.97, and 3.66 for diets A, B, and C, respectively

  5. FeatureViewer, a BioJS component for visualization of position-based annotations in protein sequences

    OpenAIRE

    Leyla Garcia; Guy Yachdav; Maria-Jesus Martin

    2014-01-01

    Summary: FeatureViewer is a BioJS component that lays out, maps, orients, and renders position-based annotations for protein sequences. This component is highly flexible and customizable, allowing the presentation of annotations by rows, all centered, or distributed in non-overlapping tracks. It uses either lines or shapes for sites and rectangles for regions. The result is a powerful visualization tool that can be easily integrated into web applications as well as documents as it provides an...

  6. Characterization of three protein components required for functional reconstitution of the epoxide carboxylase multienzyme complex from Xanthobacter strain Py2.

    OpenAIRE

    Allen, J. R.; Ensign, S A

    1997-01-01

    Epoxide carboxylase from Xanthobacter strain Py2 catalyzes the reductant- and NAD+-dependent carboxylation of aliphatic epoxides to beta-keto acids. Epoxide carboxylase from Xanthobacter strain Py2 has been resolved from cell extracts by anion-exchange chromatography into three protein components, designated I, II, and III, that are obligately required for functional reconstitution of epoxide carboxylase activity. Component II has been purified to homogeneity on the basis of its ability to co...

  7. Components of the E. coli envelope are affected by and can react to protein over-production in the cytoplasm

    Directory of Open Access Journals (Sweden)

    Villa Riccardo

    2009-06-01

    Full Text Available Abstract Background Protein over-expression in bacteria is still the easiest, cheapest and therefore preferred way to obtain large amounts of proteins for industrial and laboratory scale preparations. Several studies emphasized the importance of understanding cellular and molecular mechanisms triggered by protein over-production in order to obtain higher yield and better quality of the recombinant product. Almost every step leading to a fully functional polypeptide has been investigated, from mRNA stability to the role of molecular chaperones, from aggregation to bottlenecks in the secretory pathway. In this context, we focused on the still poorly addressed relationship between protein production in the cytoplasm and the bacterial envelope, an active and reactive cell compartment that controls interactions with the environment and several major cellular processes. Results available to date show that the accumulation of foreign proteins in the cytoplasm induces changes in the membrane lipids and in the levels of mRNAs for some membrane proteins. However, a direct connection between membrane protein expression levels and soluble/aggregated protein accumulation in the cytoplasm has never been reported. Results By the use of a combined physiological and proteomic approach, we investigated the effects on the cell membrane of E. coli of the overexpression of two recombinant proteins, the B. cepacia lipase (BCL and the green fluorescent protein (GFP. Both polypeptides are expressed in the cytoplasm at similar levels but GFP is fully soluble whereas inactive BCL accumulates in inclusion bodies. Growth and viability of the transformed cells were tested in the presence of different drugs. We found that chloramphenycol preferentially inhibited the strain over-producing GFP while SDS was more effective when BCL inclusion bodies accumulated in the cytoplasm. In contrast, both proteins induced a similar response in the membrane proteome, i.e. increased levels

  8. Autophagy-Associated Protein SmATG12 Is Required for Fruiting-Body Formation in the Filamentous Ascomycete Sordaria macrospora.

    Science.gov (United States)

    Werner, Antonia; Herzog, Britta; Frey, Stefan; Pöggeler, Stefanie

    2016-01-01

    In filamentous fungi, autophagy functions as a catabolic mechanism to overcome starvation and to control diverse developmental processes under normal nutritional conditions. Autophagy involves the formation of double-membrane vesicles, termed autophagosomes that engulf cellular components and bring about their degradation via fusion with vacuoles. Two ubiquitin-like (UBL) conjugation systems are essential for the expansion of the autophagosomal membrane: the UBL protein ATG8 is conjugated to the lipid phosphatidylethanolamine and the UBL protein ATG12 is coupled to ATG5. We recently showed that in the homothallic ascomycete Sordaria macrospora autophagy-related genes encoding components of the conjugation systems are required for fruiting-body development and/or are essential for viability. In the present work, we cloned and characterized the S. macrospora (Sm)atg12 gene. Two-hybrid analysis revealed that SmATG12 can interact with SmATG7 and SmATG3. To examine its role in S. macrospora, we replaced the open reading frame of Smatg12 with a hygromycin resistance cassette and generated a homokaryotic ΔSmatg12 knockout strain, which displayed slower vegetative growth under nutrient starvation conditions and was unable to form fruiting bodies. In the hyphae of S. macrospora EGFP-labeled SmATG12 was detected in the cytoplasm and as punctate structures presumed to be phagophores or phagophore assembly sites. Delivery of EGFP-labelled SmATG8 to the vacuole was entirely dependent on SmATG12. PMID:27309377

  9. Autophagy-Associated Protein SmATG12 Is Required for Fruiting-Body Formation in the Filamentous Ascomycete Sordaria macrospora.

    Directory of Open Access Journals (Sweden)

    Antonia Werner

    Full Text Available In filamentous fungi, autophagy functions as a catabolic mechanism to overcome starvation and to control diverse developmental processes under normal nutritional conditions. Autophagy involves the formation of double-membrane vesicles, termed autophagosomes that engulf cellular components and bring about their degradation via fusion with vacuoles. Two ubiquitin-like (UBL conjugation systems are essential for the expansion of the autophagosomal membrane: the UBL protein ATG8 is conjugated to the lipid phosphatidylethanolamine and the UBL protein ATG12 is coupled to ATG5. We recently showed that in the homothallic ascomycete Sordaria macrospora autophagy-related genes encoding components of the conjugation systems are required for fruiting-body development and/or are essential for viability. In the present work, we cloned and characterized the S. macrospora (Smatg12 gene. Two-hybrid analysis revealed that SmATG12 can interact with SmATG7 and SmATG3. To examine its role in S. macrospora, we replaced the open reading frame of Smatg12 with a hygromycin resistance cassette and generated a homokaryotic ΔSmatg12 knockout strain, which displayed slower vegetative growth under nutrient starvation conditions and was unable to form fruiting bodies. In the hyphae of S. macrospora EGFP-labeled SmATG12 was detected in the cytoplasm and as punctate structures presumed to be phagophores or phagophore assembly sites. Delivery of EGFP-labelled SmATG8 to the vacuole was entirely dependent on SmATG12.

  10. Soy versus whey protein bars: Effects on exercise training impact on lean body mass and antioxidant status

    OpenAIRE

    Babaknia Ari; DiSilvestro Robert A; Brown Erin C; Devor Steven T

    2004-01-01

    Abstract Background Although soy protein may have many health benefits derived from its associated antioxidants, many male exercisers avoid soy protein. This is due partly to a popular, but untested notion that in males, soy is inferior to whey in promoting muscle weight gain. This study provided a direct comparison between a soy product and a whey product. Methods Lean body mass gain was examined in males from a university weight training class given daily servings of micronutrient-fortified...

  11. Body composition and grip strength are improved in transgenic sickle mice fed a high-protein diet

    OpenAIRE

    Capers, Patrice L; Hyacinth, Hyacinth I; Cue, Shayla; Chappa, Prasanthi; Vikulina, Tatyana; Roser-Page, Susanne; Weitzmann, M. Neale; Archer, David R.; Newman, Gale W.; Quarshie, Alexander; Stiles, Jonathan K; Hibbert, Jacqueline M

    2015-01-01

    Key pathophysiology of sickle cell anaemia includes compensatory erythropoiesis, vascular injury and chronic inflammation, which divert amino acids from tissue deposition for growth/weight gain and muscle formation. We hypothesised that sickle mice maintained on an isoenergetic diet with a high percentage of energy derived from protein (35 %), as opposed to a standard diet with 20 % of energy derived from protein, would improve body composition, bone mass and grip strength. Male Berkeley tran...

  12. Chemical treatment of Escherichia coli: 3. Selective extraction of a recombinant protein from cytoplasmic inclusion bodies in intact cells.

    Science.gov (United States)

    Falconer, R J; O'Neill, B K; Middelberg, A P

    1999-02-20

    In previous parts of this study we developed procedures for the high-efficiency chemical extraction of soluble and insoluble protein from intact Escherichia coli cells. Although high yields were obtained, extraction of recombinant protein directly from cytoplasmic inclusion bodies led to low product purity due to coextraction of soluble contaminants. In this work, a two-stage procedure for the selective extraction of recombinant protein at high efficiency and high purity is reported. In the first stage, inclusion-body stability is promoted by the addition of 15 mM 2-hydroxyethyldisulfide (2-HEDS), also known as oxidized beta-mercaptoethanol, to the permeabilization buffer (6 M urea + 3 mM ethylenediaminetetraacetate [EDTA]). 2-HEDS is an oxidizing agent believed to promote disulfide bond formation, rendering the inclusion body resistant to solubilization in 6 M urea. Contaminating proteins are separated from the inclusion-body fraction by centrifugation. In the second stage, disulfide bonds are readily eliminated by including reducing agent (20 mM dithiothreitol [DTT]) into the permeabilization buffer. Extraction using this selective two-stage process yielded an 81% (w/w) recovery of the recombinant protein Long-R3-IGF-I from inclusion bodies located in the cytoplasm of intact E. coli, at a purity of 46% (w/w). This was comparable to that achieved by conventional extraction (mechanical disruption followed by centrifugation and solubilization). A pilot-scale procedure was also demonstrated using a stirred reactor and diafiltration. This is the first reported study that achieves both high extraction efficiency and selectivity by the chemical treatment of cytoplasmic inclusion bodies in intact bacterial cells. PMID:9921154

  13. EFFECTS OF CORDYCEPS SINENSIS PREPARATION ON BODY PROTEIN AND AMINO ACID METABOLISM IN PATIENTS AND RATS WITH CHRONIC RENAL FAILURE

    Institute of Scientific and Technical Information of China (English)

    朱淳; 刘强; 左静南; 朱汉威; 马济民

    2002-01-01

    Objective To study the effects of Cordyceps sinensis (CS) on the metabolism of body protein and intra-extracellular amino acids in patients with chronic renal failure( CRF) , and on the rates of protein synthesis in rats with CRF. Methods In patients with CRF, free amino acid concentrations in plasma and skeletal muscle before and after CS treatment were measured by the LKB-4400 amino acid automatic analytical instrument and the changes of body protein metabolism were observed by the method of 15 N-labeled glycine.Meanwhile, the rates of protein synthesis in liver ( SL % /d ) and muscle (SM%/d) of rats with CRF were determinedd by 3f-phenylalanine radioactive tracer. Results After patients with CRF were treated by CS, the Leu, lie, Thr , Lys, Cys, Tyr concentrations in plasma approached the normal levels. In one sample of skeletal muscle the Thr and Lys concentrations approached the normal, whereas both the intracellular and extracellular Val concentrations were still remarkably decreased as compared with the normal controls. Moreover, the nitrogen flow rate (Q) , rates of protein synthesis (S) and catabolism ( C) , and amino nitrogen utilization ratio (S/Q) in patients with CRF and the SL % /d and SM%/d in rats with CRF were significantly increased as compared with those before CS treatment. Conclusion CS can notably improve the amino acid metabolism, promote the body protein synthesis in patients with CRF , and increase the rates of SL % /d and SM%/d in rats with CRF.

  14. Effect of dairy proteins on appetite, energy expenditure, body weight, and composition

    DEFF Research Database (Denmark)

    Bendtsen, Line Quist; Lorenzen, Janne Kunchel; Bendsen, Nathalie Tommerup;

    2013-01-01

    proteins exert different metabolic effects. In the Western world, dairy protein, which consists of 80% casein and 20% whey, is a large contributor to our daily protein intake. Casein and whey differ in absorption and digestion rates, with casein being a "slow" protein and whey being a "fast" protein. In....... However, data indicate that amino acid composition, rate of absorption, and protein/food texture may be important factors for protein-stimulated metabolic effects....

  15. Genomic Analysis of Two-Component Signal Transduction Proteins in Basidiomycetes

    DEFF Research Database (Denmark)

    Ussery, David; Lavín, JL; Binnewies, Tim Terence;

    2010-01-01

    proteins. Several TCS proteins are highly conserved among all the basidiomycetes, and other TCS proteins appear to be specific to particular species of basidiomycetes. Moreover, some species appear to have developed a unique histidine kinase group with unusual domain architecture, the Dual-histidine...

  16. Comparative genomic analysis of two-component regulatory proteins in Pseudomonas syringae

    DEFF Research Database (Denmark)

    Lavin, J.L.; Kiil, Kristoffer; Resano, O.;

    2007-01-01

    1448A were found to contain a large number of genes encoding TCS proteins, and a core of complete TCS proteins were shared between these genomes: 30 putative TCS clusters, 11 orphan HKs, 33 orphan RRs, and 16 hybrid HKs. A close analysis of the distribution of genes encoding TCS proteins revealed...

  17. Refolding and purification of recombinant L-asparaginase from inclusion bodies of E. coli in to active tetrameric protein

    Directory of Open Access Journals (Sweden)

    AmulyaKumarPanda

    2014-09-01

    Full Text Available A tetrameric protein of therapeutic importance, Escherichia coli L-Asparaginase-II was expressed in Escherichia coli as inclusion bodies. Asparaginase inclusion bodies were solubilized using low concentration of urea and refolded into active tetrameric protein using pulsatile dilution method. Refolded asparaginase was purified in two steps using ion-exchange and gel filtration chromatographic techniques. The recovery of bioactive asparaginase from inclusion bodies was around 50 %. The melting temperature (Tm of the purified asparaginase was found to be 64 °C. The specific activity of refolded, purified asparaginase was found to be comparable to the commercial asparaginase (190 U/mg. Enzymatic activity of the refolded asparaginase was high even at four molar urea solutions, where the inclusion body aggregates are completely solubilized. From the comparison of chemical denaturation data and activity at different concentrations of guanidine hydrochloride, it was observed that dissociation of monomeric units precedes the complete loss of helical secondary structures. Protection of the existing native-like protein structure during solubilization of inclusion body aggregates with 4 M urea improved the propensity of monomer units to form oligomeric structure. This helped in improved recovery of asparaginase in bioactive tetrameric form.

  18. The Drosophila melanogaster Cajal body

    OpenAIRE

    Liu, Ji-Long; Murphy, Christine; Buszczak, Michael; Clatterbuck, Sarah; Goodman, Robyn; Gall, Joseph G.

    2006-01-01

    Cajal bodies (CBs) are nuclear organelles that are usually identified by the marker protein p80-coilin. Because no orthologue of coilin is known in Drosophila melanogaster, we identified D. melanogaster CBs using probes for other components that are relatively diagnostic for CBs in vertebrate cells. U85 small CB–specific RNA, U2 small nuclear RNA, the survival of motor neurons protein, and fibrillarin occur together in a nuclear body that is closely associated with the nucleolus. Based on its...

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

    DEFF Research Database (Denmark)

    Bendtsen, Line Quist; Lorenzen, Janne Kunchel; Larsen, Thomas Meinert;

    2014-01-01

    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 ...

  20. C-reactive protein levels and body mass index: elucidating direction of causation through reciprocal Mendelian randomization

    DEFF Research Database (Denmark)

    Timpson, N J; Nordestgaard, B G; Harbord, R M; Zacho, J; Frayling, T M; Tybjærg-Hansen, Anne; Smith, G D; Smith, G Davey

    2011-01-01

    Context:The assignment of direction and causality within networks of observational associations is problematic outside randomized control trials, and the presence of a causal relationship between body mass index (BMI) and C-reactive protein (CRP) is disputed.Objective:Using reciprocal Mendelian...

  1. Aerobic fitness level does not modulate changes in whole-body protein turnover produced by unaccustomed increases in energy expenditure

    Science.gov (United States)

    The effects of a sudden increase in energy expenditure (EE) on whole-body protein turnover vary between studies, and the possibility that fitness level modulates those responses has not been fully investigated. We hypothesized that aerobically trained individuals may exhibit adaptations that protec...

  2. Maternal low protein diet causes body weight loss in male, neonate Sprague-Dawley rats involving UCP-1 mediated thermogenesis

    Science.gov (United States)

    Brown adipose tissue (BAT) plays an important role in regulating body weight (BW) by modifying thermogenesis. Maternal low protein (LP) diets reduce offspring birth weight. Increased BAT thermogenesis in utero may be one mechanism for the lower BW. However, whether maternal LP nutrition alters BAT...

  3. Disulfide bond formation and folding of plant peroxidases expressed as inclusion body protein in Escherichia coli thioredoxin reductase negative strains

    DEFF Research Database (Denmark)

    Teilum, K; Ostergaard, L; Welinder, K G

    1999-01-01

    Escherichia coli is widely used for the production of proteins, which are of interest in structure and function studies. The folding yield of inclusion body protein is, however, generally low (a few percent) for proteins such as the plant and fungal peroxidases, which contain four disulfide bonds......, two Ca2+ ions, and a heme group. We have studied the expression yield and folding efficiency of (i) a novel Arabidopsis thaliana peroxidase, ATP N; and (ii) barley grain peroxidase, BP 1. The expression yield ranges from 0 to 60 microgram/ml of cell culture depending on the peroxidase gene and the...

  4. Composition of globoid crystals from embryo protein bodies in five species of cucurbita.

    Science.gov (United States)

    Lott, J N; Vollmer, C M

    1979-02-01

    Previous energy-dispersive x-ray analysis studies of globoid crystal composition in seed protein bodies gave an indication that there might be a correlation between seed size and the type of elements stored in globoid crystals. This possibility was tested by conducting energy-dispersive x-ray analysis studies of P, K, Mg, and Ca levels in globoid crystals of four embryo regions (radicle, stem, cotyledon center palisade mesophyll, cotyledon center spongy mesophyII) in each of five different Cucurbita species (C. mixta, C. moschata, C. foetidissima, C. pepo, and C. andreana). The species were chosen to provide a range of seed size and weight. Globoid crystals from all embryo regions in all five species contained P, K, and Mg. Some variations in the levels of these elements did occur but there was no consistent pattern with regard to area of the seed or with regard to seed size. Calcium distribution showed significant variations. In species with large seeds (C. mixta, C. moschata) Ca was mainly found in globoid crystals in the radicle. Globoid crystals in species with small seeds (C. foetidissima, C. pepo, C. andreana) contained Ca in all embryo regions tested. The results of this study support the concept that Ca distribution in globoid crystals can be correlated with seed weight. PMID:16660719

  5. Composition of Globoid Crystals from Embryo Protein Bodies in Five Species of Cucurbita1

    Science.gov (United States)

    Lott, John N. A.; Vollmer, Catherine M.

    1979-01-01

    Previous energy-dispersive x-ray analysis studies of globoid crystal composition in seed protein bodies gave an indication that there might be a correlation between seed size and the type of elements stored in globoid crystals. This possibility was tested by conducting energy-dispersive x-ray analysis studies of P, K, Mg, and Ca levels in globoid crystals of four embryo regions (radicle, stem, cotyledon center palisade mesophyll, cotyledon center spongy mesophyII) in each of five different Cucurbita species (C. mixta, C. moschata, C. foetidissima, C. pepo, and C. andreana). The species were chosen to provide a range of seed size and weight. Globoid crystals from all embryo regions in all five species contained P, K, and Mg. Some variations in the levels of these elements did occur but there was no consistent pattern with regard to area of the seed or with regard to seed size. Calcium distribution showed significant variations. In species with large seeds (C. mixta, C. moschata) Ca was mainly found in globoid crystals in the radicle. Globoid crystals in species with small seeds (C. foetidissima, C. pepo, C. andreana) contained Ca in all embryo regions tested. The results of this study support the concept that Ca distribution in globoid crystals can be correlated with seed weight. PMID:16660719

  6. Protein Supplementation with Low Fat Meat after Resistance Training: Effects on Body Composition and Strength

    Directory of Open Access Journals (Sweden)

    Massimo Negro

    2014-08-01

    Full Text Available Beef is a nutrient-rich, high-quality protein containing all the essential amino acids in proportions similar to those found in human skeletal muscle. In order to investigate the efficacy of a beef supplementation strategy on strength and body composition, we recruited 26 young healthy adults to participate in a resistance-training program of eight weeks, based on the use of isotonic machines and free weights at 75% of one repetition maximum. Subjects were randomly divided into two groups, food group and control group, of 12 and 14 subjects respectively. Food group were supplemented after resistance training with a 135 g serving of lean beef (tinned meat, providing 20 g of protein and 1.7 g of fat. No supplementation was provided to control group. Fat mass, fat free mass, lean mass, assessed by bioelectrical impedance analyzer, and muscle strength, assessed by one repetition maximum test, were evaluated in all subjects both at the beginning (week 0 and at the end (week 8 of the study. Pre- and post-training differences were evaluated with paired t-tests while group differences for each outcome parameter was evaluated with independent t-tests. At the end of the study the food group showed a significantly decrease in fat mass (week 0: 15.0 ± 6.7 kg; week 8: 13.1 ± 7.6 kg; Δ: −1.9 ± 2.9 kg; p < 0.05 and a significantly increase in fat free mass (week 0: 52.8 kg ± 9.4; week 8: 55.1 kg ± 10.9; Δ: 2.3 ± 2.5 kg; p < 0.01. No significant differences in lean mass were found in either food group or control group. No significant differences in one repetition maximum tests were found between food group and control group. Tinned meat can be considered a nutrition strategy in addition to other proteins or amino acid supplements, but as with any other supplementation strategy, a proper nutrition plan must be coupled.

  7. Study of interaction between protein and main active components in Citrus aurantium L. by optical spectroscopy

    International Nuclear Information System (INIS)

    The interaction between flavonoids and proteins was investigated by fluorescence and absorption spectroscopy. The binding parameters of drugs with proteins were obtained according to the corrected fluorescence data by an improved calculation method. The ΔH, ΔS and ΔG obtained indicate that the van der Waals or hydrogen bond, electrostatic force and hydrophobic forces all play a role in the interaction of drugs with proteins. Based on Foerster's theory, the binding average distance r between the protein and drug was evaluated and found to be less than 3 nm. The interaction of drug-metal ion complexes and proteins was also investigated.

  8. Study of interaction between protein and main active components in Citrus aurantium L. by optical spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Sun Yantao [College of Chemistry, Jilin University, Changchun 130012 (China); College of Chemistry, Jilin Normal University, Siping 136000 (China); Zhang Haitao; Sun Ye; Zhang Yupu; Liu He [College of Chemistry, Jilin University, Changchun 130012 (China); Cheng Jianhua [College of Environmental and Resources, Jilin University, Changchun 130012 (China); Bi Shuyun [College of Chemistry, Changchun Normal University, Changchun 130032 (China); Zhang Hanqi, E-mail: analchem@jlu.edu.c [College of Chemistry, Jilin University, Changchun 130012 (China)

    2010-02-15

    The interaction between flavonoids and proteins was investigated by fluorescence and absorption spectroscopy. The binding parameters of drugs with proteins were obtained according to the corrected fluorescence data by an improved calculation method. The DELTAH, DELTAS and DELTAG obtained indicate that the van der Waals or hydrogen bond, electrostatic force and hydrophobic forces all play a role in the interaction of drugs with proteins. Based on Foerster's theory, the binding average distance r between the protein and drug was evaluated and found to be less than 3 nm. The interaction of drug-metal ion complexes and proteins was also investigated.

  9. Whey protein but not soy protein supplementation alters body weight and composition in free-living overweight and obese adults

    Science.gov (United States)

    The most satiating macronutrient appears to be dietary protein; the source of the dietary protein may influence satiety and food intake. Few long-term clinical trials have investigated the effects of whey protein on biomarkers of satiety, such as plasma ghrelin, in overweight and obese adults. The e...

  10. Spirulina non-protein components induce BDNF gene transcription via HO-1 activity in C6 glioma cells.

    Science.gov (United States)

    Morita, Kyoji; Itoh, Mari; Nishibori, Naoyoshi; Her, Song; Lee, Mi-Sook

    2015-01-01

    Blue-green algae are known to contain biologically active proteins and non-protein substances and considered as useful materials for manufacturing the nutritional supplements. Particularly, Spirulina has been reported to contain a variety of antioxidants, such as flavonoids, carotenoids, and vitamin C, thereby exerting their protective effects against the oxidative damage to the cells. In addition to their antioxidant actions, polyphenolic compounds have been speculated to cause the protection of neuronal cells and the recovery of neurologic function in the brain through the production of brain-derived neurotrophic factor (BDNF) in glial cells. Then, the protein-deprived extract was prepared by removing the most part of protein components from aqueous extract of Spirulina platensis, and the effect of this extract on BDNF gene transcription was examined in C6 glioma cells. Consequently, the protein-deprived extract was shown to cause the elevation of BDNF mRNA levels following the expression of heme oxygenase-1 (HO-1) in the glioma cells. Therefore, the non-protein components of S. platensis are considered to stimulate BDNF gene transcription through the HO-1 induction in glial cells, thus proposing a potential ability of the algae to indirectly modulate the brain function through the glial cell activity. PMID:25349086

  11. A Comparing of Body Composition Components in Physically Active and Inactive Male Students of Sharoud University of Technology

    OpenAIRE

    Mehdi Broghani; mohamadreza hamedinea; mohamadreza asad; mehdi zarae

    2010-01-01

    Introduction: The aim of this study is to investigate the body mass indexes and physical activities of 11-14 year -old adolescent boys in Khooshab.Methods: This cross-sectional study was performed on 300 male students aging 11-14 years old in Khooshab (2009). Subjects were selected via random sampling. Underweight, overweight and obesity were defined based on the 5th, 85th and 95th percentiles of body mass index (BMI) for age and sex, respectively, as proposed by CDC, 2000. Physical activity ...

  12. A many-body model to study proteins. II. Incidence of many-body polarization effects on the interaction of the calmodulin protein with four Ca2+ dications and with a target enzyme peptide

    Science.gov (United States)

    Cuniasse, Philippe; Masella, Michel

    2003-07-01

    The origin of the interactions occurring in the calmodulin protein interacting with one of its target peptide and counterions, and binding four calcium dications, has been investigated in the gas phase, using the many-body model presented in Paper I [Masella and Cuniasse, J. Chem. Phys. 119, 1866 (2003)] and a classical pairwise force field. As compared to the latter force field, the many-body model is shown to provide a geometrical description of the calmodulin/target peptide structure in better agreement with the x-ray experimental one, and a better description of the Ca2+ binding sites (as compared to "small molecule" structures reported in the Cambridge Structural Database). Regarding the energy, both models provide qualitatively a similar description of the interactions occurring in the calmodulin/target peptide system. However, quantitatively, the pairwise model predicts interaction energies greater by about 25% as compared to the many-body one in the case of calmodulin/Ca2+ interactions. This is due to the inability of pairwise force fields to account for the strong anticooperative effects predicted to occur in [Ca,(carboxylate)n]2-n systems by both the many-body model and quantum computations. Hence, the new many-body model appears to be well suited for describing proteinic systems interacting with cations, both in terms of geometry and energy.

  13. Comparative study between N-body and Fokker-Planck simulations for rotating star clusters - II. 2-component models

    CERN Document Server

    Hong, Jongsuk; Lee, Hyung Mok; Spurzem, Rainer

    2012-01-01

    To understand the effects of the initial rotation on the evolution of the tidally limited clusters with mass spectrum, we have performed N-body simulations of the clusters with different initial rotations and compared the results with those of the Fokker-Planck (FP) simulations. We confirmed that the cluster evolution is accelerated by not only the initial rotation but also the mass spectrum. For the slowly rotating models, the time evolutions of mass, energy and angular momentum show good agreements between N-body and FP simulations. On the other hand, for the rapidly rotating models, there are significant differences between these two approaches at the early stage of the evolutions because of the development of bar instability in N-body simulations. The shape of the cluster for N-body simulations becomes tri-axial or even prolate, which cannot be produced by the 2-dimensional FP simulations. The total angular momentum and the total mass of the cluster decrease rapidly while bar-like structure persists. Afte...

  14. Oxa1p, an essential component of the N-tail protein export machinery in mitochondria

    OpenAIRE

    Hell, Kai; Herrmann, Johannes M.; Pratje, Elke; Neupert, Walter; Stuart, Rosemary A.

    1998-01-01

    A number of nuclear encoded inner membrane proteins of mitochondria span the membrane in such a manner that their N termini are located in the intermembrane space. Many of these proteins attain this membrane orientation by undergoing an export step from the matrix across the inner membrane. This export process, which resembles bacterial N-tail export from energetic and topogenic signal requirements, is facilitated by Oxa1p, a protein that has homologues throughout prokaryotes and eukaryotes. ...

  15. Urine proteomics: the present and future of measuring urinary protein components in disease

    OpenAIRE

    Barratt, Jonathan; Topham, Peter

    2007-01-01

    For centuries, physicians have attempted to use the urine for noninvasive assessment of disease. Today, urinalysis, in particular the measurement of proteinuria, underpins the routine assessment of patients with renal disease. More sophisticated methods for assessing specific urinary protein losses have emerged; however, albumin is still the principal urinary protein measured. Changes in the pattern of urinary protein excretion are not necessarily restricted to nephrourological disease; for i...

  16. The epsins define a family of proteins that interact with components of the clathrin coat and contain a new protein module

    DEFF Research Database (Denmark)

    Rosenthal, J A; Chen, H; Slepnev, V I;

    1999-01-01

    with the binding of both proteins to Eps15. Epsin 2, like epsin 1, is enriched in brain, is present in a brain-derived clathrin-coated vesicle fraction, is concentrated in the peri-Golgi region and at the cell periphery of transfected cells, and partially colocalizes with clathrin. High overexpression of green...... fluorescent protein-epsin 2 mislocalizes components of the clathrin coat and inhibits clathrin-mediated endocytosis. The epsins define a new protein family implicated in membrane dynamics at the cell surface.......Epsin (epsin 1) is an interacting partner for the EH domain-containing region of Eps15 and has been implicated in conjunction with Eps15 in clathrin-mediated endocytosis. We report here the characterization of a similar protein (epsin 2), which we have cloned from human and rat brain libraries...

  17. 单纯母乳喂养乳汁成分含量与产妇体重指数的相关性研究%The correlation research between milk component content in exclusive breastfeeding andmaternal body mass index

    Institute of Scientific and Technical Information of China (English)

    杨雅莉

    2015-01-01

    Objective To evaluate correlation research between milk component content in exclusive breastfeeding andmaternal body mass index.Methods To investigate and analyze the maternal age, height, body weight before childbirth way, prenatal, gestational age, birth weight, baby weight, gender, height in 140 case. milk component content in exclusive breastfeeding andmaternal body mass index were contrastive analysised.Results Antenatal and pre-pregnancy BMI satisfied the equation BMI = 9.036 + 0.873 *. Maternal body index increases were positively associated with milk composition of zinc, calcium, magnesium, protein, fat, minerals, and the solid fat, were negatively correlated with iron, copper, and lactose.Conclusion Maternal body index increases were correlated with milk component content;controlling the maternal weight increase can keep the balance of milk component content.%目的:探讨研究单纯母乳喂养乳汁成分含量与产妇体重指数的相关性。方法:调查分析我院妇产科分娩的140例产妇年龄、身高、孕前体重、产前体重、孕周、分娩方式、婴儿体重、性别、身长等,对比分析产妇体重增长与乳汁成分含量的相关性。结果:产妇产前和孕前BMI满足方程:产前BMI=9.036+0.873*孕前BMI,产妇体重指数增幅与乳汁成分中锌元素、钙元素、镁元素、蛋白质、脂肪、矿物质和非脂肪固体呈正相关,与铁元素、铜元素和乳糖呈负相关。结论:产妇体重指数增加与乳汁成分含量有一定的相关性;控制产妇体重增加可保持乳汁成分含量的均衡性。

  18. Protein interactome mining defines melatonin MT1 receptors as integral component of presynaptic protein complexes of neurons.

    Science.gov (United States)

    Benleulmi-Chaachoua, Abla; Chen, Lina; Sokolina, Kate; Wong, Victoria; Jurisica, Igor; Emerit, Michel Boris; Darmon, Michèle; Espin, Almudena; Stagljar, Igor; Tafelmeyer, Petra; Zamponi, Gerald W; Delagrange, Philippe; Maurice, Pascal; Jockers, Ralf

    2016-01-01

    In mammals, the hormone melatonin is mainly produced by the pineal gland with nocturnal peak levels. Its peripheral and central actions rely either on its intrinsic antioxidant properties or on binding to melatonin MT1 and MT2 receptors, belonging to the G protein-coupled receptor (GPCR) super-family. Melatonin has been reported to be involved in many functions of the central nervous system such as circadian rhythm regulation, neurotransmission, synaptic plasticity, memory, sleep, and also in Alzheimer's disease and depression. However, little is known about the subcellular localization of melatonin receptors and the molecular aspects involved in neuronal functions of melatonin. Identification of protein complexes associated with GPCRs has been shown to be a valid approach to improve our understanding of their function. By combining proteomic and genomic approaches we built an interactome of MT1 and MT2 receptors, which comprises 378 individual proteins. Among the proteins interacting with MT1 , but not with MT2 , we identified several presynaptic proteins, suggesting a potential role of MT1 in neurotransmission. Presynaptic localization of MT1 receptors in the hypothalamus, striatum, and cortex was confirmed by subcellular fractionation experiments and immunofluorescence microscopy. MT1 physically interacts with the voltage-gated calcium channel Cav 2.2 and inhibits Cav 2.2-promoted Ca(2+) entry in an agonist-independent manner. In conclusion, we show that MT1 is part of the presynaptic protein network and negatively regulates Cav 2.2 activity, providing a first hint for potential synaptic functions of MT1. PMID:26514267

  19. Endotoxin-free purification for the isolation of Bovine Viral Diarrhoea Virus E2 protein from insoluble inclusion body aggregates

    Directory of Open Access Journals (Sweden)

    Mahony Timothy J

    2011-07-01

    Full Text Available Abstract Background Protein expression in Escherichia coli may result in the recombinant protein being expressed as insoluble inclusion bodies. In addition, proteins purified from E. coli contain endotoxins which need to be removed for in vivo applications. The structural protein, E2, from Bovine Viral Diarrhoea Virus (BVDV is a major immunogenic determinant, and is an ideal candidate as a subunit vaccine. The E2 protein contains 17 cysteine residues creating difficulties in E. coli expression. In this report we outline a procedure for successfully producing soluble and endotoxin-free BVDV E2 protein from inclusion bodies (IB. Results The expression of a truncated form of BVDV-E2 protein (E2-T1 in E. coli resulted in predominantly aggregated insoluble IB. Solubilisation of E2-T1 with high purity and stability from IB aggregates was achieved using a strong reducing buffer containing 100 mM Dithiothreitol. Refolding by dialysis into 50 mM Tris (pH 7.0 containing 0.2% Igepal CA630 resulted in a soluble but aggregated protein solution. The novel application of a two-phase extraction of inclusion body preparations with Triton X-114 reduced endotoxin in solubilised E2-T1 to levels suitable for in vivo use without affecting protein yields. Dynamic light scattering analyses showed 37.5% of the protein was monomeric, the remaining comprised of soluble aggregates. Mice immunised with E2-T1 developed a high titre antibody response by ELISA. Western hybridisation analysis showed E2-T1 was recognised by sera from immunised mice and also by several BVDV-E2 polyclonal and monoclonal antibodies. Conclusion We have developed a procedure using E. coli to produce soluble E2-T1 protein from IB, and due to their insoluble nature we utilised a novel approach using Triton X-114 to efficiently remove endotoxin. The resultant protein is immunogenic and detectable by BVDV-E2 specific antibodies indicating its usefulness for diagnostic applications and as a subunit

  20. Body composition and grip strength are improved in transgenic sickle mice fed a high-protein diet.

    Science.gov (United States)

    Capers, Patrice L; Hyacinth, Hyacinth I; Cue, Shayla; Chappa, Prasanthi; Vikulina, Tatyana; Roser-Page, Susanne; Weitzmann, M Neale; Archer, David R; Newman, Gale W; Quarshie, Alexander; Stiles, Jonathan K; Hibbert, Jacqueline M

    2015-01-01

    Key pathophysiology of sickle cell anaemia includes compensatory erythropoiesis, vascular injury and chronic inflammation, which divert amino acids from tissue deposition for growth/weight gain and muscle formation. We hypothesised that sickle mice maintained on an isoenergetic diet with a high percentage of energy derived from protein (35 %), as opposed to a standard diet with 20 % of energy derived from protein, would improve body composition, bone mass and grip strength. Male Berkeley transgenic sickle mice (S; n 8-12) were fed either 20 % (S20) or 35 % (S35) diets for 3 months. Grip strength (BIOSEB meter) and body composition (dual-energy X-ray absorptiometry scan) were measured. After 3 months, control mice had the highest bone mineral density (BMD) and bone mineral content (BMC) (P gain, BMD, BMC and lean body mass compared with other groups. TS3.2 mice showed significantly more improvement in grip strength than TS0·8 and TS1.6 mice (P < 0·05). In conclusion, the high-protein diet improved body composition and grip strength. Outcomes observed with TS1.6 and TS3.2 mice, respectively, confirm the hypothesis and reveal l-Arg as part of the mechanism. PMID:26090102

  1. Enhancement of protein secretion in Saccharomyces cerevisiae by overproduction of Sso protein, a late-acting component of the secretory machinery.

    Science.gov (United States)

    Ruohonen, L; Toikkanen, J; Tieaho, V; Outola, M; Soderlund, H; Keranen, S

    1997-03-30

    Increased production of secreted proteins in Saccharomyces cerevisiae was achieved by overexpressing the yeast syntaxins. Sso1 or Sso2 protein, the t-SNAREs functioning at the targeting/fusion of the Golgi-derived secretory vesicles to the plasma membrane. Up to four- or six-fold yields of a heterologous secreted protein, Bacillus alpha-amylase, or an endogenous secreted protein, invertase, were obtained respectively when expressing either one of the SSO genes, SSO1 or SSO2, from the ADH1 promoter on a multicopy plasmid. Direct correlation between the Sso protein level and the amount of secreted alpha-amylase was demonstrated by modulating the expression level of the SSO2 gene. Quantitation of the alpha-amylase activity in the culture medium, periplasmic space and cytoplasm suggests that secretion into the periplasmic space is the primary stage at which the SSO genes exert the secretion-enhancing function. Pulse-chase data also support enhanced secretion efficiently obtained by SSO overexpression. Our data suggest that the Sso proteins may be rate-limiting components of the protein secretion machinery at the exocytosis step in yeast. PMID:9133737

  2. Proteomic analysis of secreted protein induced by a component of prey in pitcher fluid of the carnivorous plant Nepenthes alata.

    Science.gov (United States)

    Hatano, Naoya; Hamada, Tatsuro

    2012-08-01

    The Nepenthes species are carnivorous plants that have evolved a specialized leaf organ, the 'pitcher', to attract, capture, and digest insects. The digested insects provide nutrients for growth, allowing these plants to grow even in poor soil. Several proteins have been identified in the pitcher fluid, including aspartic proteases (nepenthesin I and II) and pathogenesis-related (PR) proteins (β-1,3-glucanase, class IV chitinase, and thaumatin-like protein). In this study, we collected and concentrated pitcher fluid to identify minor proteins. In addition, we tried to identify the protein secreted in response to trapping the insect. To make a similar situation in which the insect falls into the pitcher, chitin which was a major component of the insect exoskeleton was added to the fluid in the pitcher. Three PR proteins, class III peroxidase (Prx), β-1,3-glucanase, and class III chitinase, were newly identified. Prx was induced after the addition of chitin to the pitcher fluid. Proteins in the pitcher fluid of the carnivorous plant Nepenthes alata probably have two roles in nutrient supply: digestion of prey and the antibacterial effect. These results suggest that the system for digesting prey has evolved from the defense system against pathogens in the carnivorous plant Nepenthes. PMID:22705321

  3. Conditional QTL mapping of protein content in wheat with respect to grain yield and its components

    Indian Academy of Sciences (India)

    Lin Wang; Fa Cui; Jinping Wang; Li Jun; Anming Ding; Chunhua Zhao; Xingfeng Li; Deshun Feng; Jurong Gao; Honggang Wang

    2012-12-01

    Grain protein content in wheat (Triticum aestivum L.) is generally considered a highly heritable character that is negatively correlated with grain yield and yield-related traits. Quantitative trait loci (QTL) for protein content was mapped using data on protein content and protein content conditioned on the putatively interrelated traits to evaluate possible genetic interrelationships between protein content and yield, as well as yield-related traits. Phenotypic data were evaluated in a recombinant inbred line population with 302 lines derived from a cross between the Chinese cultivar Weimai 8 and Luohan 2. Inclusive composite interval mapping using IciMapping 3.0 was employed for mapping unconditional and conditional QTL with additives. A strong genetic relationship was found between protein content and grain yield, and yield-related traits. Unconditional QTL mapping analysis detected seven additive QTL for protein content, with additive effects ranging in absolute size from 0.1898% to 0.3407% protein content, jointly accounting for 43.45% of the trait variance. Conditional QTL mapping analysis indicated two QTL independent from yield, which can be used in marker-assisted selection for increasing yield without affecting grain protein content. Three additional QTL with minor effects were identified in the conditional mapping. Of the three QTLs, two were identified when protein content was conditioned on yield, which had pleiotropic effects on those two traits. Conditional QTL mapping can be used to dissect the genetic interrelationship between two traits at the individual QTL level for closely correlated traits. Further, conditional QTL mapping can reveal additional QTL with minor effects that are undetectable in unconditional mapping.

  4. RELATION BETWEEN THE COMPONENT OF BODY FITNESS AND HAPPINESS AND LIFE QUALITY OF NEYSHABURE AZAD UNIVERSITY GIRL STUDENT

    OpenAIRE

    Amir Moghadam; Soheila Goharrokhi

    2011-01-01

    Today life quality is index of social development .this index includes the most importantdifferent factors that determine conditions of life in society and personal ease of life .For thedevelopment of society all factors of life quality should be promoted one by one on that. So; thedetermination of significant life quality factors & the ways of promotion are the necessities ofsociety development. One of the effective factors in life quality is body fitness.So; this paperaims to examine the re...

  5. Mms19 protein functions in nucleotide excision repair by sustaining an adequate cellular concentration of the TFIIH component Rad3

    OpenAIRE

    Kou, Haiping; Ying ZHOU; Gorospe, R.M. Charlotte; Wang, Zhigang

    2008-01-01

    Nucleotide excision repair (NER) is a major cellular defense mechanism against DNA damage. We have investigated the role of Mms19 in NER in the yeast Saccharomyces cerevisiae. NER was deficient in the mms19 deletion mutant cell extracts, which was complemented by the NER/transcription factor TFIIH, but not by purified Mms19 protein. In mms19 mutant cells, protein levels of the core TFIIH component Rad3 (XPD homologue) and Ssl2 (XPB homologue) were significantly reduced by up to 3.5- and 2.2-f...

  6. Group component protein-derived macrophage activating factor stimulates macrophages that induce human breast cancer cell apoptosis.

    OpenAIRE

    Ruggiero, M; L. Thyer; E. Wards; Smith, R; J.J.V. Branca; Gulisano, M; G.Morucci; S. Pacini

    2013-01-01

    Objectives: The vitamin D axis is involved in various aspects of human breast cancer. It is composed by vitamin D, vitamin D receptor (VDR) and group component (Gc) protein that is the precursor of the Gc protein-derived macrophage activating factor (GcMAF) (Eur Nephrol. 2011;5(1):15–9). It was demonstrated that administration of GcMAF to metastatic breast cancer patients yielded good clinical results (Int J Cancer. 2008 Jan 15;122(2):461-7). Here we demonstrate that GcMAF stimulates ma...

  7. Proteomic analysis of pure human airway gland mucus reveals a large component of protective proteins.

    Directory of Open Access Journals (Sweden)

    Nam Soo Joo

    Full Text Available Airway submucosal glands contribute to innate immunity and protect the lungs by secreting mucus, which is required for mucociliary clearance and which also contains antimicrobial, anti-inflammatory, anti-proteolytic and anti-oxidant proteins. We stimulated glands in tracheal trimmings from three lung donors and collected droplets of uncontaminated mucus as they formed at the gland orifices under an oil layer. We analyzed the mucus using liquid chromatography-tandem mass spectrometry (LC-MS/MS. Analysis identified 5486 peptides and 441 proteins from across the 3 samples (269-319 proteins per subject. We focused on 269 proteins common to at least 2 0f 3 subjects, of which 102 (38% had protective or innate immunity functions. While many of these have long been known to play such roles, for many others their cellular protective functions have only recently been appreciated in addition to their well-studied biologic functions (e.g. annexins, apolipoproteins, gelsolin, hemoglobin, histones, keratins, and lumican. A minority of the identified proteins are known to be secreted via conventional exocytosis, suggesting that glandular secretion occurs via multiple mechanisms. Two of the observed protective proteins, major vault protein and prohibitin, have not been observed in fluid from human epithelial cultures or in fluid from nasal or bronchoalveolar lavage. Further proteomic analysis of pure gland mucus may help clarify how healthy airways maintain a sterile environment.

  8. DEXA measures of body fat percentage and acute phase proteins among breast cancer survivors: a Cross-Sectional Analysis

    International Nuclear Information System (INIS)

    C-reactive protein (CRP) and Serum amyloid A protein (SAA) increases with systemic inflammation and are related to worse survival for breast cancer survivors. This study examines the association between percent body fat and SAA and CRP and the potential interaction with NSAID use and weight change. Participants included 134 non-Hispanic white and Hispanic breast cancer survivors from the Health, Eating, Activity, and Lifestyle Study. Body fat percentage, measured with Dual Energy X-ray Absorptiometer (DEXA), and circulating levels of CRP and SAA were obtained 30 months after breast cancer diagnosis. Circulating concentrations of CRP and SAA were associated with increased adiposity as measured by DEXA after adjustment for age at 24-months, race/ethnicity, dietary energy intake, weight change, and NSAID use. Survivors with higher body fat ≥35% had significantly higher concentrations of CRP (2.01 mg/l vs. 0.85 mg/l) and SAA (6.21 mg/l vs. 4.21 mg/l) compared to non-obese (body fat < 35%). Women who had gained more than 5% of their body weight since breast cancer diagnosis had non-statistically significant higher geometric mean levels of CRP and SAA. Mean levels of CRP and SAA were higher among obese women who were non-users of NSAIDs compared to current users; the association with SAA reached statistical significance (Mean SAA = 7.24, 95%CI 6.13-8.56 for non-NSAID; vs. 4.87; 95%CI 3.95-6.0 for NSAID users respectively). Breast cancer survivors with higher body fat had higher mean concentrations of CRP and SAA than women with lower body fat. Further assessment of NSAID use and weight control in reducing circulating inflammatory markers among survivors may be worthwhile to investigate in randomized intervention trials as higher inflammatory markers are associated with worse survival

  9. Kinetic analysis of the reactions of hypobromous acid with protein components

    DEFF Research Database (Denmark)

    Pattison, David I; Davies, Michael Jonathan

    2004-01-01

    proteins isolated from patients with atherosclerosis, asthma, and cystic fibrosis, implicating the production of HOX in these diseases. The quantitative significance of these findings requires knowledge of the kinetics of reaction of HOX with protein targets, and such data have not been previously...... more, and Cys and Met much less, important targets for HOBr than HOCl. Kinetic models have been developed to predict the targets of HOX attack on proteins and free amino acids. Overall, these results shed light on the mechanisms of cell damage induced by HOX and indicate, for example, that the 3-chloro...

  10. The response of muscle protein synthesis following whole-body resistance exercise is greater following 40 g than 20 g of ingested whey protein.

    Science.gov (United States)

    Macnaughton, Lindsay S; Wardle, Sophie L; Witard, Oliver C; McGlory, Chris; Hamilton, D Lee; Jeromson, Stewart; Lawrence, Clare E; Wallis, Gareth A; Tipton, Kevin D

    2016-08-01

    The currently accepted amount of protein required to achieve maximal stimulation of myofibrillar protein synthesis (MPS) following resistance exercise is 20-25 g. However, the influence of lean body mass (LBM) on the response of MPS to protein ingestion is unclear. Our aim was to assess the influence of LBM, both total and the amount activated during exercise, on the maximal response of MPS to ingestion of 20 or 40 g of whey protein following a bout of whole-body resistance exercise. Resistance-trained males were assigned to a group with lower LBM (≤65 kg; LLBM n = 15) or higher LBM (≥70 kg; HLBM n = 15) and participated in two trials in random order. MPS was measured with the infusion of (13)C6-phenylalanine tracer and collection of muscle biopsies following ingestion of either 20 or 40 g protein during recovery from a single bout of whole-body resistance exercise. A similar response of MPS during exercise recovery was observed between LBM groups following protein ingestion (20 g - LLBM: 0.048 ± 0.018%·h(-1); HLBM: 0.051 ± 0.014%·h(-1); 40 g - LLBM: 0.059 ± 0.021%·h(-1); HLBM: 0.059 ± 0.012%·h(-1)). Overall (groups combined), MPS was stimulated to a greater extent following ingestion of 40 g (0.059 ± 0.020%·h(-1)) compared with 20 g (0.049 ± 0.020%·h(-1); P = 0.005) of protein. Our data indicate that ingestion of 40 g whey protein following whole-body resistance exercise stimulates a greater MPS response than 20 g in young resistance-trained men. However, with the current doses, the total amount of LBM does not seem to influence the response. PMID:27511985

  11. Arsenic mediated disruption of promyelocytic leukemia protein nuclear bodies induces ganciclovir susceptibility in Epstein-Barr positive epithelial cells

    International Nuclear Information System (INIS)

    Promyelocytic leukemia protein nuclear bodies (PML NBs) have been implicated in host immune response to viral infection. PML NBs are targeted for degradation during reactivation of herpes viruses, suggesting that disruption of PML NB function supports this aspect of the viral life cycle. The Epstein-Barr virus (EBV) Latent Membrane Protein 1 (LMP1) has been shown to suppress EBV reactivation. Our finding that LMP1 induces PML NB immunofluorescence intensity led to the hypothesis that LMP1 may modulate PML NBs as a means of maintaining EBV latency. Increased PML protein and morphometric changes in PML NBs were observed in EBV infected alveolar epithelial cells and nasopharyngeal carcinoma cells. Treatment with low dose arsenic trioxide disrupted PML NBs, induced expression of EBV lytic proteins, and conferred ganciclovir susceptibility. This study introduces an effective modality to induce susceptibility to ganciclovir in epithelial cells with implications for the treatment of EBV associated pathologies.

  12. Interactions of Streptococcus mutans Fimbria-Associated Surface Proteins with Salivary Components

    OpenAIRE

    Ray, Chad A.; Gfell, Linda E.; Buller, Tiffany L.; Gregory, Richard L

    1999-01-01

    Streptococcus mutans has been implicated as the major causative agent of human dental caries. S. mutans binds to saliva-coated tooth surfaces, and previous studies suggested that fimbriae may play a role in the initial bacterial adherence to salivary components. The objectives of this study were to establish the ability of an S. mutans fimbria preparation to bind to saliva-coated surfaces and determine the specific salivary components that facilitate binding with f...

  13. Flavoproteins, iron proteins, and hemoproteins as electron-transfer components of the sulfate-reducing bacteria

    Energy Technology Data Exchange (ETDEWEB)

    LeGall, J. (Centre National de la Recherche Scientifique, Marseille, France); DerVartanian, D.V.; Peck, H.D. Jr.

    1979-01-01

    This review article with 105 references discusses the most recent publications that deal with the discovery of new redox proteins of the sulfate-reducing bacteria belonging to the genera Desulfotomaculum and Desulfovibrio and proposes explanations for their physical and biological functions. The redox proteins studied as part of the electron-transfer system of these bacteria include flavodoxins, ferredoxins, rubredoxins, cytochromes and several reductose-type enzymes. (KRM)

  14. Body composition, dietary composition, and components of metabolic syndrome in overweight and obese adults after a 12-week trial on dietary treatments focused on portion control, energy density, or glycemic index

    Directory of Open Access Journals (Sweden)

    Melanson Kathleen J

    2012-08-01

    Full Text Available Abstract Background Given the rise in obesity and associated chronic diseases, it is critical to determine optimal weight management approaches that will also improve dietary composition and chronic disease risk factors. Few studies have examined all these weight, diet, and disease risk variables in subjects participating in recommended multi-disciplinary weight loss programs using different dietary strategies. Methods This study compared effects of three dietary approaches to weight loss on body composition, dietary composition and risk factors for metabolic syndrome (MetS. In a 12-week trial, sedentary but otherwise healthy overweight and obese adults (19 M & 138 F; 38.7 ± 6.7 y; BMI 31.8 ± 2.2 who were attending weekly group sessions for weight loss followed either portion control, low energy density, or low glycemic index diet plans. At baseline and 12 weeks, measures included anthropometrics, body composition, 3-day food diaries, blood pressure, total lipid profile, HOMA, C-reactive protein, and fasting blood glucose and insulin. Data were analyzed by repeated measures analysis of variance. Results All groups significantly reduced body weight and showed significant improvements in body composition (p  Conclusion Different dietary approaches based on portion control, low energy density, or low glycemic index produced similar, significant short-term improvements in body composition, diet compositin, and MetS components in overweight and obese adults undergoing weekly weight loss meetings. This may allow for flexibility in options for dietary counseling based on patient preference.

  15. A Comparing of Body Composition Components in Physically Active and Inactive Male Students of Sharoud University of Technology

    Directory of Open Access Journals (Sweden)

    Mehdi Broghani

    2010-01-01

    Full Text Available Introduction: The aim of this study is to investigate the body mass indexes and physical activities of 11-14 year -old adolescent boys in Khooshab.Methods: This cross-sectional study was performed on 300 male students aging 11-14 years old in Khooshab (2009. Subjects were selected via random sampling. Underweight, overweight and obesity were defined based on the 5th, 85th and 95th percentiles of body mass index (BMI for age and sex, respectively, as proposed by CDC, 2000. Physical activity levels were assessed by the International Physical Activity Questionnaire (IPAQ. Sedentary behaviors and socioeconomic status of subjects were assessed by means of a researcher- made questionnaire.Results: The overall prevalence of underweight and overweight-obesity was 16% and 3.38%, respectively. The levels of light; intermediate, and heavy physical activity among the obese subjects were 19, 24 and 15, among normal subjects were 19, 44, and 36, and among the lean ones were 22, 60 and 41 mins in 24 hours. Physical activity levels significantly were lower (P<0.01 in overweight-obese subjects than in normal-weight and underweight subjects. Television watching times were higher (P<0.05 in overweight-obese subjects than normal-weight subjects. Education levels of parents were significantly higher in overweight-obese subjects than normal-weight subjects.Conclusion: we concluded that prevalence of underweight was higher than prevalence overweight and obesity among 11-14 year-old adolescent boys in Khooshab. Furthermore, physical activity was not related to body mass index and increase in sedentary behaviors (television waticng were related with obesity and overweight in our subjects.

  16. Dietary n-6 PUFA, carbohydrate:protein ratio and change in body weight and waist circumference

    DEFF Research Database (Denmark)

    Jakobsen, Marianne U; Madsen, Lise; Dethlefsen, Claus;

    2015-01-01

    -order terms, protein intake, long-chain n-3 PUFA intake and other potential confounders. Due to adjustment for intake of protein, levels of carbohydrate indirectly reflect levels of the carbohydrate:protein ratio. SETTING: Diet, Cancer and Health follow-up study, Denmark. SUBJECTS: Women and men (n 29 152...

  17. Characterization of a novel snake venom component: Kazal-type inhibitor-like protein from the arboreal pitviper Bothriechis schlegelii.

    Science.gov (United States)

    Fernández, Julián; Gutiérrez, José María; Calvete, Juan J; Sanz, Libia; Lomonte, Bruno

    2016-06-01

    Snake venoms are composed mainly of a mixture of proteins and peptides. Notably, all snake venom toxins have been assigned to a small number of protein families. Proteomic studies on snake venoms have recently identified the presence of Kazal-type inhibitor-like proteins in the neotropical arboreal snakes Bothriechis schlegelii and Bothriechis supraciliaris. In the present study, a Kazal-type component from B. schlegelii, named Kazal-type inhibitor-like protein (KTIL), has been completely sequenced and characterized for the first time. This protein, which contains 54 amino acid residues, shows sequence similarity to the third domain of the ovomucoid from avian species, which is a Kazal-like domain. KTIL did not inhibit the enzymatic activity of various serine proteinases at pH = 7.2 or pH = 8.0, but partially inhibited the activity of trypsin at pH = 5.4, and the only toxic effect in mice observed after different in vivo tests was the induction of footpad edema. KTIL was not lethal when injected in mice or chickens. The presence of Kazal-type proteins and mRNA only in species of the genus Bothriechis suggests a genus recruitment event in the early-Middle Miocene, the estimated time of emergence of this clade. PMID:26973135

  18. A study of whole body protein kinetics in malnourished children with persistent diarrhea: A double blind trial of zinc supplementation

    International Nuclear Information System (INIS)

    Persistent diarrhoea (PD) is globally recognized as a major cause of childhood morbidity and mortality. PD is closely associated with malnutrition and nutrition rehabilitation especially domiciliary dietary therapy posses a therapeutic challenge. While there has been success in managing such children with locally home available traditional diets, there has been concern with the potential of associated micronutrient especially zinc deficiency. We are evaluating the impact of zinc supplementation of a traditional rice-lentil (khitchri) and yogurt diet in malnourished children with PD in randomized double blind study. In addition to the impact on weight gain, stool output and body composition, we will evaluate whole body protein kinetics using the modified CRP protocol [employing 15N-glycine, H13Co3 and 1-13C leucine]. We will also estimate the effect of coexisting illnesses, especially febrile episodes on nutritional recovery and protein metabolism. (author). 20 refs, 5 figs, 1 tab

  19. Toxic and nontoxic components of botulinum neurotoxin complex are evolved from a common ancestral zinc protein

    Energy Technology Data Exchange (ETDEWEB)

    Inui, Ken [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Japan Society for the Promotion of Science, 1-8 Chiyoda-ku, Tokyo 102-8472 (Japan); Sagane, Yoshimasa [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Miyata, Keita [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Japan Society for the Promotion of Science, 1-8 Chiyoda-ku, Tokyo 102-8472 (Japan); Miyashita, Shin-Ichiro [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Suzuki, Tomonori [Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8558 (Japan); Shikamori, Yasuyuki [Agilent Technologies International Japan, Ltd. Takaura-cho 9-1, Hachioji-shi, Tokyo 192-0033 (Japan); Ohyama, Tohru; Niwa, Koichi [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan); Watanabe, Toshihiro, E-mail: t-watana@bioindustry.nodai.ac.jp [Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493 (Japan)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer BoNT and NTNHA proteins share a similar protein architecture. Black-Right-Pointing-Pointer NTNHA and BoNT were both identified as zinc-binding proteins. Black-Right-Pointing-Pointer NTNHA does not have a classical HEXXH zinc-coordinating motif similar to that found in all serotypes of BoNT. Black-Right-Pointing-Pointer Homology modeling implied probable key residues involved in zinc coordination. -- Abstract: Zinc atoms play an essential role in a number of enzymes. Botulinum neurotoxin (BoNT), the most potent toxin known in nature, is a zinc-dependent endopeptidase. Here we identify the nontoxic nonhemagglutinin (NTNHA), one of the BoNT-complex constituents, as a zinc-binding protein, along with BoNT. A protein structure classification database search indicated that BoNT and NTNHA share a similar domain architecture, comprising a zinc-dependent metalloproteinase-like, BoNT coiled-coil motif and concanavalin A-like domains. Inductively coupled plasma-mass spectrometry analysis demonstrated that every single NTNHA molecule contains a single zinc atom. This is the first demonstration of a zinc atom in this protein, as far as we know. However, the NTNHA molecule does not possess any known zinc-coordinating motif, whereas all BoNT serotypes possess the classical HEXXH motif. Homology modeling of the NTNHA structure implied that a consensus K-C-L-I-K-X{sub 35}-D sequence common among all NTNHA serotype molecules appears to coordinate a single zinc atom. These findings lead us to propose that NTNHA and BoNT may have evolved distinct functional specializations following their branching out from a common ancestral zinc protein.

  20. Toxic and nontoxic components of botulinum neurotoxin complex are evolved from a common ancestral zinc protein

    International Nuclear Information System (INIS)

    Highlights: ► BoNT and NTNHA proteins share a similar protein architecture. ► NTNHA and BoNT were both identified as zinc-binding proteins. ► NTNHA does not have a classical HEXXH zinc-coordinating motif similar to that found in all serotypes of BoNT. ► Homology modeling implied probable key residues involved in zinc coordination. -- Abstract: Zinc atoms play an essential role in a number of enzymes. Botulinum neurotoxin (BoNT), the most potent toxin known in nature, is a zinc-dependent endopeptidase. Here we identify the nontoxic nonhemagglutinin (NTNHA), one of the BoNT-complex constituents, as a zinc-binding protein, along with BoNT. A protein structure classification database search indicated that BoNT and NTNHA share a similar domain architecture, comprising a zinc-dependent metalloproteinase-like, BoNT coiled-coil motif and concanavalin A-like domains. Inductively coupled plasma-mass spectrometry analysis demonstrated that every single NTNHA molecule contains a single zinc atom. This is the first demonstration of a zinc atom in this protein, as far as we know. However, the NTNHA molecule does not possess any known zinc-coordinating motif, whereas all BoNT serotypes possess the classical HEXXH motif. Homology modeling of the NTNHA structure implied that a consensus K-C-L-I-K-X35-D sequence common among all NTNHA serotype molecules appears to coordinate a single zinc atom. These findings lead us to propose that NTNHA and BoNT may have evolved distinct functional specializations following their branching out from a common ancestral zinc protein.

  1. Interaction between genetic predisposition to adiposity and dietary protein in relation to subsequent change in body weight and waist circumference.

    Directory of Open Access Journals (Sweden)

    Mikkel Z Ankarfeldt

    Full Text Available Genetic predisposition to adiposity may interact with dietary protein in relation to changes of anthropometry.To investigate the interaction between genetic predisposition to higher body mass index (BMI, waist circumference (WC or waist-hip ratio adjusted for BMI (WHRBMI and dietary protein in relation to subsequent change in body weight (ΔBW or change in WC (ΔWC.Three different Danish cohorts were used. In total 7,054 individuals constituted the study population with information on diet, 50 single-nucleotide polymorphisms (SNPs associated with BMI, WC or WHRBMI, as well as potential confounders. Mean follow-up time was ∼5 years. Four genetic predisposition-scores were based on the SNPs; a complete-score including all selected adiposity- associated SNPs, and three scores including BMI, WC or WHRBMI associated polymorphisms, respectively. The association between protein intake and ΔBW or ΔWC were examined and interactions between SNP-score and protein were investigated. Analyses were based on linear regressions using macronutrient substitution models and meta-analyses.When protein replaced carbohydrate, meta-analyses showed no associations with ΔBW (41.0 gram/y/5 energy% protein, [95% CI: -32.3; 114.3] or ΔWC (<-0.1 mm/y/5 energy % protein, [-1.1; 1.1]. Similarly, there were no interactions for any SNP-scores and protein for either ΔBW (complete SNP-score: 1.8 gram/y/5 energy% protein/risk allele, [-7.0; 10.6] or ΔWC (complete SNP-score: <0.1 mm/y/5 energy% protein/risk allele, [-0.1; 0.1]. Similar results were seen when protein replaced fat.This study indicates that the genetic predisposition to general and abdominal adiposity, assessed by gene-scores, does not seem to modulate the influence of dietary protein on ΔBW or ΔWC.

  2. Signal-dependent Regulation of Transcription by Histone Deacetylase 7 Involves Recruitment to Promyelocytic Leukemia Protein Nuclear Bodies

    OpenAIRE

    Gao, Chengzhuo; Cheng, Xiwen; Lam, Minh; Yu LIU; Liu, Qing; Chang, Kun-Sang; Kao, Hung-Ying

    2008-01-01

    Promyelocytic leukemia protein (PML) nuclear bodies (NBs) are dynamic subnuclear compartments that play roles in several cellular processes, including apoptosis, transcriptional regulation, and DNA repair. Histone deacetylase (HDAC) 7 is a potent corepressor that inhibits transcription by myocyte enhancer factor 2 (MEF2) transcription factors. We show here that endogenous HDAC7 and PML interact and partially colocalize in PML NBs. Tumor necrosis factor (TNF)-α treatment recruits HDAC7 to PML ...

  3. Induction of alternative lengthening of telomeres-associated PML bodies by p53/p21 requires HP1 proteins

    OpenAIRE

    Jiang, Wei-Qin; Zhong, Ze-Huai; Nguyen, Akira; Henson, Jeremy D.; Toouli, Christian D.; Braithwaite, Antony W.; Reddel, Roger R

    2009-01-01

    Alternative lengthening of telomeres (ALT) is a recombination-mediated process that maintains telomeres in telomerase-negative cancer cells. In asynchronously dividing ALT-positive cell populations, a small fraction of the cells have ALT-associated promyelocytic leukemia nuclear bodies (APBs), which contain (TTAGGG)n DNA and telomere-binding proteins. We found that restoring p53 function in ALT cells caused p21 up-regulation, growth arrest/senescence, and a large increase in cells containing ...

  4. Phase Transitions in the Nucleus: the functional implications of concentration-dependent assembly of a Liquid-like RNA/Protein Body

    Science.gov (United States)

    Zhu, Lian; Weber, Stephanie; Berry, Joel; Vaidya, Nilesh; Haataja, Mikko; Brangwynne, Clifford

    2015-03-01

    The nucleolus is a liquid-like membrane-less nuclear body which plays an important role in cell growth and size control. By modulating nucleolar component concentration through RNAi conditions that change C. elegans cell size, we find that nucleoli only assemble above a threshold concentration; moreover, the ripening dynamics of nucleated droplets are consistent with the hypothesis that the assembly of the nucleolus represents an intracellular liquid-liquid phase transition. A key question is how this phase-transition is linked to the primary function of the nucleolus, in transcribing and processing ribosomal RNA. To address this, we characterize the localization of RNA Polymerase I, a key transcriptional enzyme, into nucleolar foci as a function of nucleolar component concentration. Our results suggest that there are a small number of key disordered phosphoproteins that may serve as a link between transcription and assembly. Finally, we present preliminary results using a reduced model system consisting of purified nucleolar proteins to assess the ability of nucleolar proteins to drive liquid-liquid phase separation in vitro. These results lay the foundation for a quantitative understanding of intracellular phase transitions and their impact on biomedically-critical RNA-processing steps.

  5. Early origin and adaptive evolution of the GW182 protein family, the key component of RNA silencing in animals.

    Science.gov (United States)

    Zielezinski, Andrzej; Karlowski, Wojciech M

    2015-01-01

    The GW182 proteins are a key component of the miRNA-dependent post-transcriptional silencing pathway in animals. They function as scaffold proteins to mediate the interaction of Argonaute (AGO)-containing complexes with cytoplasmic poly(A)-binding proteins (PABP) and PAN2-PAN3 and CCR4-NOT deadenylases. The AGO-GW182 complexes mediate silencing of the target mRNA through induction of translational repression and/or mRNA degradation. Although the GW182 proteins are a subject of extensive experimental research in the recent years, very little is known about their origin and evolution. Here, based on complex functional annotation and phylogenetic analyses, we reveal 448 members of the GW182 protein family from the earliest animals to humans. Our results indicate that a single-copy GW182/TNRC6C progenitor gene arose with the emergence of multicellularity and it multiplied in the last common ancestor of vertebrates in 2 rounds of whole genome duplication (WGD) resulting in 3 genes. Before the divergence of vertebrates, both the AGO- and CCR4-NOT-binding regions of GW182s showed significant acceleration in the accumulation of amino acid changes, suggesting functional adaptation toward higher specificity to the molecules of the silencing complex. We conclude that the silencing ability of the GW182 proteins improves with higher position in the taxonomic classification and increasing complexity of the organism. The first reconstruction of the molecular journey of GW182 proteins from the ancestral metazoan protein to the current mammalian configuration provides new insight into development of the miRNA-dependent post-transcriptional silencing pathway in animals. PMID:26106978

  6. Yeast mitochondrial HMG proteins: DNA-binding properties of the most evolutionarily divergent component of mitochondrial nucleoids.

    Science.gov (United States)

    Bakkaiova, Jana; Marini, Victoria; Willcox, Smaranda; Nosek, Jozef; Griffith, Jack D; Krejci, Lumir; Tomaska, Lubomir

    2016-01-01

    Yeast mtDNA is compacted into nucleoprotein structures called mitochondrial nucleoids (mt-nucleoids). The principal mediators of nucleoid formation are mitochondrial high-mobility group (HMG)-box containing (mtHMG) proteins. Although these proteins are some of the fastest evolving components of mt-nucleoids, it is not known whether the divergence of mtHMG proteins on the level of their amino acid sequences is accompanied by diversification of their biochemical properties. In the present study we performed a comparative biochemical analysis of yeast mtHMG proteins from Saccharomyces cerevisiae (ScAbf2p), Yarrowia lipolytica (YlMhb1p) and Candida parapsilosis (CpGcf1p). We found that all three proteins exhibit relatively weak binding to intact dsDNA. In fact, ScAbf2p and YlMhb1p bind quantitatively to this substrate only at very high protein to DNA ratios and CpGcf1p shows only negligible binding to dsDNA. In contrast, the proteins exhibit much higher preference for recombination intermediates such as Holliday junctions (HJ) and replication forks (RF). Therefore, we hypothesize that the roles of the yeast mtHMG proteins in maintenance and compaction of mtDNA in vivo are in large part mediated by their binding to recombination/replication intermediates. We also speculate that the distinct biochemical properties of CpGcf1p may represent one of the prerequisites for frequent evolutionary tinkering with the form of the mitochondrial genome in the CTG-clade of hemiascomycetous yeast species. PMID:26647378

  7. A SAS-6-like protein suggests that the Toxoplasma conoid complex evolved from flagellar components

    OpenAIRE

    De Leon, Jessica; Scheumann, Nicole; Beatty, Wandy; Beck, Josh R; Tran, Johnson Q.; Yau, Candace; Bradley, Peter J.; Gull, Keith; Wickstead, Bill; Morrissette, Naomi S.

    2013-01-01

    SAS-6 is required for centriole biogenesis in diverse eukaryotes. Here, we describe a novel family of SAS-6-like (SAS6L) proteins that share an N-terminal domain with SAS-6 but lack coiled-coil tails. SAS6L proteins are found in a subset of eukaryotes that contain SAS-6, including diverse protozoa and green algae. In the apicomplexan parasite Toxoplasma gondii, SAS-6 localizes to the centriole but SAS6L is found above the conoid, an enigmatic tubulin-containing structure found at the apex of ...

  8. Preventing galvanic corrosion - Systematic development of a magnesium car body component; Vermeidung von Bimetallkorrosion - Systematische Entwicklung eines Magnesium Karosseriebauteils

    Energy Technology Data Exchange (ETDEWEB)

    Schreckenberger, H.; Izquierdo, P.; Klose, S.G. [Daimler AG, Stuttgart (Germany); Heitmann, Volker; Hoeche, D.; Kainer, K.U. [GKSS, Geesthacht (Germany); Blawert, C.

    2010-10-15

    The hatch back of the new Mercedes-Benz E-Class T-Model is one example for a hybrid design with inner magnesium high pressure die casting liner and outer aluminium sheet. The hybrid lift gate has to fulfil a variety of requirements corresponding to dimensional stability, weight, stiffness, crash and corrosion performance. The hybrid concept in general as well as various interfaces and connecting points require a careful design to prevent galvanic corrosion. Critical points are introduced and for two examples (gas pressure spring and hinge) the constructive criterions are discussed. Finally, the concept of corrosion protection measures and results of various testing trials of the component are presented.

  9. Neuronal apoptosis and neurofilament protein expression in the lateral geniculate body of cats following acute optic nerve injuries

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    BACKGROUND: The visual pathway have 6 parts, involving optic nerve, optic chiasm, optic tract, lateral geniculate body, optic radiation and cortical striatum area. Corresponding changes may be found in these 6 parts following optic nerve injury. At present, studies mainly focus on optic nerve and retina, but studies on lateral geniculate body are few.OBJECTIVE: To prepare models of acute optic nerve injury for observing the changes of neurons in lateral geniculate body, expression of neurofilament protein at different time after injury and cell apoptosis under the optical microscope, and for investigating the changes of neurons in lateral geniculate body following acute optic nerve injury.DESIGN: Completely randomized grouping design, controlled animal experiment.SETTING: Department of Neurosurgery, General Hospital of Ji'nan Military Area Command of Chinese PLA.MATERIALS: Twenty-eight adult healthy cats of either gender and common grade, weighing from 2.0 to 3.5 kg, were provided by the Animal Experimental Center of Fudan University. The involved cats were divided into 2 groups according to table of random digit: normal control group (n =3) and model group (n =25). Injury 6 hours, 1, 3, 7 and 14 days five time points were set in model group for later observation, 5 cats at each time point. TUNEL kit (Bohringer-Mannheim company)and NF200& Mr 68 000 mouse monoclonal antibody (NeoMarkers Company) were used in this experiment.METHODS: This experiment was carried out in the Department of Neurosurgery, General Hospital of Ji'nan Military Area Command of Chinese PLA between June 2004 and June 2005. ① The cats of model group were developed into cat models of acute intracranial optic nerve injury as follows: The anesthetized cats were placed in lateral position. By imitating operation to human, pterion approach was used. An incision was made at the joint line between outer canthus and tragus, and deepened along cranial base until white optic nerve via optic nerve pore

  10. Bioactive protein-based nanofibers interact with intestinal biological components resulting in transepithelial permeation of a therapeutic protein

    DEFF Research Database (Denmark)

    Boutrup Stephansen, Karen; García-Díaz, María; Jessen, Flemming;

    2015-01-01

    FSP as a biomaterial facilitated interactions with cells and enzymes found in the gastrointestinal tract and displayed excellent biocompatibility. More specifically, insulin was efficiently encapsulated into FSP fibers maintaining its conformation, and subsequent controlled release was obtained in...... simulated intestinal fluid. The encapsulation of insulin into FSP fibers provided protection against chymotrypsin degradation, and resulted in an increase in insulin transport to around 12% without compromising the cellular viability. This increased transport was driven by interactions upon contact between......Proteins originating from natural sources may constitute a novel type of material for use in drug delivery. However, thorough understanding of the behavior and effects of such a material when processed into a matrix together with a drug is crucial prior to further development into a drug product...

  11. Numerical simulation of two-dimensional heat transfer in composite bodies with application to de-icing of aircraft components. Ph.D. Thesis. Final Report

    Science.gov (United States)

    Chao, D. F. K.

    1983-01-01

    Transient, numerical simulations of the de-icing of composite aircraft components by electrothermal heating were performed for a two dimensional rectangular geometry. The implicit Crank-Nicolson formulation was used to insure stability of the finite-difference heat conduction equations and the phase change in the ice layer was simulated using the Enthalpy method. The Gauss-Seidel point iterative method was used to solve the system of difference equations. Numerical solutions illustrating de-icer performance for various composite aircraft structures and environmental conditions are presented. Comparisons are made with previous studies. The simulation can also be used to solve a variety of other heat conduction problems involving composite bodies.

  12. Binding of a cementum attachment protein to extracellular matrix components and to dental surfaces

    International Nuclear Information System (INIS)

    Cementum proteins (CP) have been shown to mediate cell attachment. Among these, a 55 kDa protein was isolated. The purpose of the present study was to assess the capacity of CP to bind to non-demineralized and demineralized root surfaces and to support cell attachment to dentin. CP were prepared by sequential extraction of bovine cementum with 25 mM EDTA, 0.5 M acetic acid followed by 4 M guanidine HCl. The latter was subjected to ion exchange chromatography on a DEAE-3SW column and eluted stepwise with a 0-0.5 M NaCl gradient. CP were labelled with 125I and the capacity of 125I-CP to bind to mineralized and partially demineralized dentin, synthetic hydroxyapatite, collagen, fibronectin and fibrillar collagen-fibronectin cimplex was assessed. It was found that CP bind specifically to mineralized dentin and synthetic hydroxyapatite but not to demineralized dentin. The specific binding was 60% of the total binding. SDS-PAGE analysis of the proteins bound to dentin indicated that the main bound protein had a molecular weight of 55 kDa. CP exhibited high affinity for fibronectin (kD = 1.56 x 10-10 M) and fibronectincollagen complex, but their binding to either molecular or fibrillar collagen was negligible. It is suggested that CP may play an important role in the attachment of cells of the periodontium to cementum extracellular matrix during homeostasis and regeneration. (au)

  13. SUMOylation regulates the nuclear mobility of CREB binding protein and its association with nuclear bodies in live cells

    Energy Technology Data Exchange (ETDEWEB)

    Ryan, Colm M.; Kindle, Karin B.; Collins, Hilary M. [Gene Regulation Group, Centre for Biomolecular Sciences, School of Pharmacy, University of Nottingham, University Park, Nottingham NG7 2RD (United Kingdom); Heery, David M., E-mail: david.heery@nottingham.ac.uk [Gene Regulation Group, Centre for Biomolecular Sciences, School of Pharmacy, University of Nottingham, University Park, Nottingham NG7 2RD (United Kingdom)

    2010-01-01

    The lysine acetyltransferase CREB binding protein (CBP) is required for chromatin modification and transcription at many gene promoters. In fixed cells, a large proportion of CBP colocalises to PML or nuclear bodies. Using live cell imaging, we show here that YFP-tagged CBP expressed in HEK293 cells undergoes gradual accumulation in nuclear bodies, some of which are mobile and migrate towards the nuclear envelope. Deletion of a short lysine-rich domain that contains the major SUMO acceptor sites of CBP abrogated its ability to be SUMO modified, and prevented its association with endogenous SUMO-1/PML speckles in vivo. This SUMO-defective CBP showed enhanced ability to co-activate AML1-mediated transcription. Deletion mapping revealed that the SUMO-modified region was not sufficient for targeting CBP to PML bodies, as C-terminally truncated mutants containing this domain showed a strong reduction in accumulation at PML bodies. Fluorescence recovery after photo-bleaching (FRAP) experiments revealed that YFP-CBP{Delta}998-1087 had a retarded recovery time in the nucleus, as compared to YFP-CBP. These results indicate that SUMOylation regulates CBP function by influencing its shuttling between nuclear bodies and chromatin microenvironments.

  14. SUMOylation regulates the nuclear mobility of CREB binding protein and its association with nuclear bodies in live cells

    International Nuclear Information System (INIS)

    The lysine acetyltransferase CREB binding protein (CBP) is required for chromatin modification and transcription at many gene promoters. In fixed cells, a large proportion of CBP colocalises to PML or nuclear bodies. Using live cell imaging, we show here that YFP-tagged CBP expressed in HEK293 cells undergoes gradual accumulation in nuclear bodies, some of which are mobile and migrate towards the nuclear envelope. Deletion of a short lysine-rich domain that contains the major SUMO acceptor sites of CBP abrogated its ability to be SUMO modified, and prevented its association with endogenous SUMO-1/PML speckles in vivo. This SUMO-defective CBP showed enhanced ability to co-activate AML1-mediated transcription. Deletion mapping revealed that the SUMO-modified region was not sufficient for targeting CBP to PML bodies, as C-terminally truncated mutants containing this domain showed a strong reduction in accumulation at PML bodies. Fluorescence recovery after photo-bleaching (FRAP) experiments revealed that YFP-CBPΔ998-1087 had a retarded recovery time in the nucleus, as compared to YFP-CBP. These results indicate that SUMOylation regulates CBP function by influencing its shuttling between nuclear bodies and chromatin microenvironments.

  15. BioTAP-XL - crosslinking/tandem affinity purification to study DNA targets, RNA and protein components of chromatin associated complexes

    OpenAIRE

    Alekseyenko, Artyom A.; McElroy, Kyle A.; Kang, Hyuckjoon; Zee, Barry M.; Kharchenko, Peter V.; Kuroda, Mitzi I.

    2015-01-01

    In order to understand how chromatin complexes function in the nucleus, it is important to obtain a comprehensive picture of their protein, DNA, and RNA components and their mutual interactions. Here we present a chromatin cross-linking approach (BioTAP-XL) which utilizes mass-spectrometry to identify protein complex components, together with high-throughput sequencing to identify RNA components and DNA binding sites. We describe full protocols for Drosophila cells and for human cells in cult...

  16. Long-distance translocation of protein during morphogenesis of the fruiting body in the filamentous fungus, Agaricus bisporus.

    Directory of Open Access Journals (Sweden)

    Benjamin M Woolston

    Full Text Available Commercial cultivation of the mushroom fungus, Agaricus bisporus, utilizes a substrate consisting of a lower layer of compost and upper layer of peat. Typically, the two layers are seeded with individual mycelial inoculants representing a single genotype of A. bisporus. Studies aimed at examining the potential of this fungal species as a heterologous protein expression system have revealed unexpected contributions of the mycelial inoculants in the morphogenesis of the fruiting body. These contributions were elucidated using a dual-inoculant method whereby the two layers were differientially inoculated with transgenic β-glucuronidase (GUS and wild-type (WT lines. Surprisingly, use of a transgenic GUS line in the lower substrate and a WT line in the upper substrate yielded fruiting bodies expressing GUS activity while lacking the GUS transgene. Results of PCR and RT-PCR analyses for the GUS transgene and RNA transcript, respectively, suggested translocation of the GUS protein from the transgenic mycelium colonizing the lower layer into the fruiting body that developed exclusively from WT mycelium colonizing the upper layer. Effective translocation of the GUS protein depended on the use of a transgenic line in the lower layer in which the GUS gene was controlled by a vegetative mycelium-active promoter (laccase 2 and β-actin, rather than a fruiting body-active promoter (hydrophobin A. GUS-expressing fruiting bodies lacking the GUS gene had a bonafide WT genotype, confirmed by the absence of stably inherited GUS and hygromycin phosphotransferase selectable marker activities in their derived basidiospores and mycelial tissue cultures. Differientially inoculating the two substrate layers with individual lines carrying the GUS gene controlled by different tissue-preferred promoters resulted in up to a ∼3.5-fold increase in GUS activity over that obtained with a single inoculant. Our findings support the existence of a previously undescribed

  17. Whole body and skeletal muscle protein turnover in recovery from burns

    OpenAIRE

    Porter, Craig; Hurren, Nicholas M; Herndon, David N; Børsheim, Elisabet

    2013-01-01

    Trauma and critical illness are associated with a stress response that results in increased skeletal muscle protein catabolism, which is thought to facilitate the synthesis of acute phase proteins in the liver as well as proteins involved in immune function. What makes burn injury a unique form of trauma is the existence of vast skin lesions, where the majority of afflicted tissue is often surgically excised post injury. Thereafter, recovery is dependent on the formation of a significant quan...

  18. Electron paramagnetic resonance study of lipid and protein membrane components of erythrocytes oxidized with hydrogen peroxide

    International Nuclear Information System (INIS)

    Electron paramagnetic resonance (EPR) spectroscopy of spin labels was used to monitor membrane dynamic changes in erythrocytes subjected to oxidative stress with hydrogen peroxide (H2O2). The lipid spin label, 5-doxyl stearic acid, responded to dramatic reductions in membrane fluidity, which was correlated with increases in the protein content of the membrane. Membrane rigidity, associated with the binding of hemoglobin (Hb) to the erythrocyte membrane, was also indicated by a spin-labeled maleimide, 5-MSL, covalently bound to the sulfhydryl groups of membrane proteins. At 2% hematocrit, these alterations in membrane occurred at very low concentrations of H2O2 (50 µM) after only 5 min of incubation at 37°C in azide phosphate buffer, pH 7.4. Lipid peroxidation, suggested by oxidative hemolysis and malondialdehyde formation, started at 300 µM H2O2 (for incubation of 3 h), which is a concentration about six times higher than those detected with the probes. Ascorbic acid and α-tocopherol protected the membrane against lipoperoxidation, but did not prevent the binding of proteins to the erythrocyte membrane. Moreover, the antioxidant (+)-catechin, which also failed to prevent the cross-linking of cytoskeletal proteins with Hb, was very effective in protecting erythrocyte ghosts from lipid peroxidation induced by the Fenton reaction. This study also showed that EPR spectroscopy can be useful to assess the molecular dynamics of red blood cell membranes in both the lipid and protein domains and examine oxidation processes in a system that is so vulnerable to oxidation

  19. Electron paramagnetic resonance study of lipid and protein membrane components of erythrocytes oxidized with hydrogen peroxide

    Energy Technology Data Exchange (ETDEWEB)

    Mendanha, S.A.; Anjos, J.L.V.; Silva, A.H.M.; Alonso, A. [Instituto de Física, Universidade Federal de Goiás, Goiânia, GO (Brazil)

    2012-04-05

    Electron paramagnetic resonance (EPR) spectroscopy of spin labels was used to monitor membrane dynamic changes in erythrocytes subjected to oxidative stress with hydrogen peroxide (H{sub 2}O{sub 2}). The lipid spin label, 5-doxyl stearic acid, responded to dramatic reductions in membrane fluidity, which was correlated with increases in the protein content of the membrane. Membrane rigidity, associated with the binding of hemoglobin (Hb) to the erythrocyte membrane, was also indicated by a spin-labeled maleimide, 5-MSL, covalently bound to the sulfhydryl groups of membrane proteins. At 2% hematocrit, these alterations in membrane occurred at very low concentrations of H{sub 2}O{sub 2} (50 µM) after only 5 min of incubation at 37°C in azide phosphate buffer, pH 7.4. Lipid peroxidation, suggested by oxidative hemolysis and malondialdehyde formation, started at 300 µM H{sub 2}O{sub 2} (for incubation of 3 h), which is a concentration about six times higher than those detected with the probes. Ascorbic acid and α-tocopherol protected the membrane against lipoperoxidation, but did not prevent the binding of proteins to the erythrocyte membrane. Moreover, the antioxidant (+)-catechin, which also failed to prevent the cross-linking of cytoskeletal proteins with Hb, was very effective in protecting erythrocyte ghosts from lipid peroxidation induced by the Fenton reaction. This study also showed that EPR spectroscopy can be useful to assess the molecular dynamics of red blood cell membranes in both the lipid and protein domains and examine oxidation processes in a system that is so vulnerable to oxidation.

  20. Purification and characterization of an antibacterial protein from dried fruiting bodies of the wild mushroom Clitocybe sinopica.

    Science.gov (United States)

    Zheng, Suyue; Liu, Qinghong; Zhang, Guoqing; Wang, Hexiang; Ng, Tzi Bun

    2010-01-01

    A novel antibacterial protein with a molecular mass of 44 kDa has been isolated from dried fruiting bodies of the wild mushroom Clitocybe sinopica. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis showed that the protein was composed of two subunits each with a molecular mass of 22 kDa. Its N-terminal amino-acid sequence, SVQATVNGDKML, has not been reported for other antimicrobial proteins. The purification protocol included ion exchange chromatography on DEAE-cellulose, CM-cellulose and Q-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The antibacterial protein was adsorbed on all three ion exchangers. The antimicrobial activity profile of the protein against tested bacterial and fungal strains disclosed that it possessed potent antibacterial activity against Agrobacterium rhizogenes, A. tumefaciens, A. vitis, Xanthomonas oryzae and X. malvacearum with a minimum inhibitory concentration mostly below 0.6 microM. However, it had no antibacterial activity against Pseudomonas batatae, Erwinia herbicola, Escherichia coli, and Staphylococcus aureus, and no antifungal activity against Setosphaeria turcica, Fusarium oxysporum, Verticillium dahliae, Bipolaris maydis, and B. sativum. The antibacterial antivity against A. tumefaciens was stable after exposure to 20-60 degrees C for 30 min and to pH 4-9 for 1 h. PMID:20198215

  1. The chaperone DnaK controls the fractioning of functional protein between soluble and insoluble cell fractions in inclusion body-forming cells

    Directory of Open Access Journals (Sweden)

    Ventura Salvador

    2006-08-01

    Full Text Available Abstract Background The molecular mechanics of inclusion body formation is still far from being completely understood, specially regarding the occurrence of properly folded, protein species that exhibit natural biological activities. We have here comparatively explored thermally promoted, in vivo protein aggregation and the formation of bacterial inclusion bodies, from both structural and functional sides. Also, the status of the soluble and insoluble protein versions in both aggregation systems have been examined as well as the role of the main molecular chaperones GroEL and DnaK in the conformational quality of the target polypeptide. Results While thermal denaturation results in the formation of heterogeneous aggregates that are rather stable in composition, protein deposition as inclusion bodies renders homogenous but strongly evolving structures, which are progressively enriched in the main protein species while gaining native-like structure. Although both type of aggregates display common features, inclusion body formation but not thermal-induced aggregation involves deposition of functional polypeptides that confer biological activity to such particles, at expenses of the average conformational quality of the protein population remaining in the soluble cell fraction. In absence of DnaK, however, the activity and conformational nativeness of inclusion body proteins are dramatically impaired while the soluble protein version gains specific activity. Conclusion The chaperone DnaK controls the fractioning of active protein between soluble and insoluble cell fractions in inclusion body-forming cells but not during thermally-driven protein aggregation. This cell protein, probably through diverse activities, is responsible for the occurrence and enrichment in inclusion bodies of native-like, functional polypeptides, that are much less represented in other kind of protein aggregates.

  2. Earth moving machine whole-body vibration and the contribution of Sub-1Hz components to ISO 2631-1 metrics.

    Science.gov (United States)

    Mansfield, Neil J; Newell, Geraldine S; Notini, Luca

    2009-08-01

    Exposure to whole-body vibration (WBV) is an occupational hazard for operators of industrial vehicles, such as earth-moving machines. Quantification of WBV exposure in terms of impact on health forms one aspect of the Standard ISO 2631-1 (1997). Regarding assessment of risk to health, ISO 2631-1 (1997) states that if WBV components below 1 Hz are not ;relevant nor important' then they can be excluded from the assessment. In this paper the influence of sub-1 Hz components in WBV acquired from a sample of 46 earth moving machines is evaluated in terms of their contribution to ISO 2631-1 WBV exposure dose metrics: frequency weighted r.m.s. and the vibration dose value (VDV). For the majority of machines, a high proportion of the horizontal (x- and y-axis) WBV r.m.s. and VDV values was generated by sub-1 Hz vibration components; there was a much lower proportion of the vertical (z-axis) vibration generated by such components. PMID:19672014

  3. Plant density and mineral nitrogen fertilization influencing yield, yield components and concentration of oil and protein in soybean grains

    Directory of Open Access Journals (Sweden)

    André Sampaio Ferreira

    2016-01-01

    Full Text Available ABSTRACT There are few studies on the interaction between soybean plant density and nitrogen fertilization. This research aimed to assess the effect of mineral nitrogen associated to different plant densities on yield, yield components and oil and protein concentrations of soybean grains. Two experiments were conducted in the 2013/2014 and 2014/2015 growing seasons, with randomized complete block design, in a split plots scheme, with six replications. Four sowing densities (150, 300, 440 and 560 thousand viable seeds∙ha–1 were allocated in the plots, and two nitrogen levels (0 and 45 kg N∙ha–1, applied at V2, using ammonium sulfate were allocated in the subplots. There was no interaction between soybean plant density and the application of mineral nitrogen on yield, yield components and oil and protein concentrations in soybean grains. Higher plant population reduced the number of pods per plant and the contribution of branch sinks to the grain yield, but the effects on yield differed among the growing seasons. The mineral nitrogen fertilization did not increase yield and protein and oil concentrations in the grains, thus it was unnecessary.

  4. Discovering the hidden sub-network component in a ranked list of genes or proteins derived from genomic experiments.

    Science.gov (United States)

    García-Alonso, Luz; Alonso, Roberto; Vidal, Enrique; Amadoz, Alicia; de María, Alejandro; Minguez, Pablo; Medina, Ignacio; Dopazo, Joaquín

    2012-11-01

    Genomic experiments (e.g. differential gene expression, single-nucleotide polymorphism association) typically produce ranked list of genes. We present a simple but powerful approach which uses protein-protein interaction data to detect sub-networks within such ranked lists of genes or proteins. We performed an exhaustive study of network parameters that allowed us concluding that the average number of components and the average number of nodes per component are the parameters that best discriminate between real and random networks. A novel aspect that increases the efficiency of this strategy in finding sub-networks is that, in addition to direct connections, also connections mediated by intermediate nodes are considered to build up the sub-networks. The possibility of using of such intermediate nodes makes this approach more robust to noise. It also overcomes some limitations intrinsic to experimental designs based on differential expression, in which some nodes are invariant across conditions. The proposed approach can also be used for candidate disease-gene prioritization. Here, we demonstrate the usefulness of the approach by means of several case examples that include a differential expression analysis in Fanconi Anemia, a genome-wide association study of bipolar disorder and a genome-scale study of essentiality in cancer genes. An efficient and easy-to-use web interface (available at http://www.babelomics.org) based on HTML5 technologies is also provided to run the algorithm and represent the network. PMID:22844098

  5. The influence of complexing agent and proteins on the corrosion of stainless steels and their metal components.

    Science.gov (United States)

    Kocijan, Aleksandra; Milosev, Ingrid; Pihlar, Boris

    2003-01-01

    The present work is devoted to the problem of biodegradation of orthopaedic implants manufactured from stainless steel. In vitro simulations of the biocompatibility of two types of stainless steel, AISI 304 and AISI 316L, and their individual metal components, i.e. iron, chromium, nickel and molybdenum, were carried out in simulated physiological solution (Hank's) containing complexing agents. Knowledge of the effects of the chemical and biological complexing agents, EDTA and proteins, respectively, on the corrosion resistance of a metal should provide a better understanding of the processes occurring in vivo on its surface. The behavior of stainless steels and metal components was studied under open circuit and under potentiostatic conditions. The concentration of dissolved corrosion products in the form of released ions was determined by differential pulse polarography (DPP) and atomic emission spectrometry using inductively coupled plasma (ICP-AES). The composition of solid corrosion products formed on the surface was analyzed by energy dispersive X-ray spectroscopy (EDS) and their morphology was viewed by scanning electron microscopy (SEM). The addition of EDTA and proteins to physiological solution increased the dissolution of pure metals and stainless steels. The effect of particular protein differs on different metals and alloys. PMID:15348541

  6. Targeting and Assembly of Components of the TOC Protein Import Complex at the Chloroplast Outer Envelope Membrane

    Directory of Open Access Journals (Sweden)

    Lynn G.L. Richardson

    2014-06-01

    Full Text Available The translocon at the outer envelope membrane of chloroplasts (TOC initiates the import of thousands of nuclear encoded preproteins required for chloroplast biogenesis and function. The multimeric TOC complex contains two GTP-regulated receptors, Toc34 and Toc159, which recognize the transit peptides of preproteins and initiate protein import through a β–barrel membrane channel, Toc75. Different isoforms of Toc34 and Toc159 assemble with Toc75 to form structurally and functionally diverse translocons, and the composition and levels of TOC translocons is required for the import of specific subsets of coordinately expressed proteins during plant growth and development. Consequently, the proper assembly of the TOC complexes is key to ensuring organelle homeostasis. This review will focus on our current knowledge of the targeting and assembly of TOC components to form functional translocons at the outer membrane. Our analyses reveal that the targeting of TOC components involves elements common to the targeting of other outer membrane proteins, but also include unique features that appear to have evolved to specifically facilitate assembly of the import apparatus.

  7. Rac-1 and Raf-1 kinases, components of distinct signaling pathways, activate myotonic dystrophy protein kinase

    Science.gov (United States)

    Shimizu, M.; Wang, W.; Walch, E. T.; Dunne, P. W.; Epstein, H. F.

    2000-01-01

    Myotonic dystrophy protein kinase (DMPK) is a serine-threonine protein kinase encoded by the myotonic dystrophy (DM) locus on human chromosome 19q13.3. It is a close relative of other kinases that interact with members of the Rho family of small GTPases. We show here that the actin cytoskeleton-linked GTPase Rac-1 binds to DMPK, and coexpression of Rac-1 and DMPK activates its transphosphorylation activity in a GTP-sensitive manner. DMPK can also bind Raf-1 kinase, the Ras-activated molecule of the MAP kinase pathway. Purified Raf-1 kinase phosphorylates and activates DMPK. The interaction of DMPK with these distinct signals suggests that it may play a role as a nexus for cross-talk between their respective pathways and may partially explain the remarkable pleiotropy of DM.

  8. Vegetarian diet components, protein and blood pressure: which nutrients are important?

    Science.gov (United States)

    Beilin, L J; Burke, V

    1995-03-01

    1. Evidence that vegetarian dietary patterns lower blood pressure (BP) comes from both population studies and randomized controlled trials in normotensive and hypertensive subjects. 2. The effect has been shown most clearly in those who keep to a strict lacto-ovo vegetarian diet characterized by a relatively low intake of saturated fat, a high polyunsaturated/saturated fat ratio, and a high intake of fruit, vegetables and other fibre containing products. Randomized controlled dietary trials suggest the effects are independent of dietary sodium, additive to that of calorie restriction, and not due to the absence of meat protein per se. Indeed, recent population studies suggest an inverse relationship between dietary protein and BP. 4. Dietary fats, fibre, potassium, magnesium and calcium do not independently seem to account for the effects. A possible role for complex carbohydrate in conjunction with the other dietary factors has yet to be fully explored. PMID:7554413

  9. Plasmalemma vesicle-associated protein: A crucial component of vascular homeostasis

    Science.gov (United States)

    Guo, Ling; Zhang, Hongyan; Hou, Yinglong; Wei, Tianshu; Liu, Ju

    2016-01-01

    Endothelial subcellular structures, including caveolae, fenestrae and transendothelial channels, are crucial for regulating microvascular function. Plasmalemma vesicle-associated protein (PLVAP) is an endothelial cell-specific protein that forms the stomatal and fenestral diaphragms of blood vessels and regulates basal permeability, leukocyte migration and angiogenesis. Loss of PLVAP in mice leads to premature mortality due to disrupted homeostasis. Evidence from previous studies suggested that PLVAP is involved in cancer, traumatic spinal cord injury, acute ischemic brain disease, transplant glomerulopathy, Norrie disease and diabetic retinopathy. Specifically, PLVAP expression has been demonstrated to be upregulated in these diseases, accompanied by pro-angiogenic or pro-inflammatory responses. Therefore, PLVAP is considered a novel therapeutic target, in addition to an endothelial cell marker. The present review summarizes the structure and functions of PLVAP, and its roles in pathophysiological processes. PMID:27602081

  10. Telomere repeat binding proteins are functional components of Arabidopsis telomeres and interact with telomerase

    Czech Academy of Sciences Publication Activity Database

    Schrumpfová, P.; Vychodilová, I.; Dvořáčková, Martina; Majerská, J.; Dokládal, Ladislav; Schorová, Š.; Fajkus, Jiří

    2014-01-01

    Roč. 77, č. 5 (2014), s. 770-781. ISSN 0960-7412 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0068; GA ČR(CZ) GA13-06943S Institutional support: RVO:68081707 Keywords : telomere protein interaction * telomere repeat binding * Arabidopsis thaliana Subject RIV: BO - Biophysics Impact factor: 5.972, year: 2014

  11. Regulation of mitochondrial and nuclear effect of thyroid hormones by cytoplasmic/protein /components

    International Nuclear Information System (INIS)

    Specific T4-binding protein (T4 effect modulator, T4EM), mediating a number of significant hormone effects in nucleus and mitochondria have been discovered in liver and brain cytoplasm. It is a glycoprotein with molecular mass of 24 kDa, consisting of 200 amino acid residues with alanine in N-terminal. T4EM is revealed to bind extremely low concentration of T4 (Ka=1, 3?109M-1). In liver cells alterations in organism thyroid status have been established to effect the modulator's activity greatly. T4EM isolated from the liver of hyperthyroid rats stimulated mitochondrial protein biosynthesis significantly as compared to the modulator isolated from the lever of intact and thyroidectomized rats. The modulator's activity has been revealed to be subjected to significant variations depending not only on functional state of thyroid gland, but on animal's age as well, changing in liver and brain differently. In liver gradual increase of the modulator's activity correlating to the animal's age and reaching its maximum in 3-month rats has been observed, to be in compliance with calorigenic effect of T4 in this organ. In brain cells T4EM activity increase has been revealed in late embryonic and early postnatal period of rat life. Beside stimulation of mitochondrial protein biosynthesis in rat lever T4EM increases protein phosphorylation, nuclear RNA polymerize activity and mitochondria respiration. Similar studies on brain cells show that adult brain the modulator does not cause confident alteration in any of studied parameters. On the basis of findings T4EM is suggested to be necessary the formation of age and tissue responsiveness to thyroxine. (author)

  12. Modulation of function of multidrug resistance associated-proteins by Kaempferia parviflora extracts and their components

    OpenAIRE

    Patanasethanont, Denpong; Nagai, Junya; Matsuura, Chie; Fukui, Kyoko; Sutthanut, Khaetthareeya; Sripanidkulchai, Bung-Orn; Yumoto, Ryoko; Takano, Mikihisa

    2007-01-01

    In this study, the effects of extracts and flavone derivatives from the rhizome of Kaempferia parviflora on multidrug resistance associated-proteins (MRP)-mediated transport in A549 cells were examined. The cells employed express MRP1 and MRP2, but not P-glycoprotein. The cellular accumulation of calcein, an MRP substrate, was significantly increased by various MRP inhibitors without being affected by verapamil, a typical P-glycoprotein inhibitor. Ethanol and aqueous extracts from Kaempferia ...

  13. Capture of a protein synthesis-dependent component of long-term depression

    OpenAIRE

    Kauderer, Beth S.; Kandel, Eric R.

    2000-01-01

    Hippocampal-based behavioral memories and hippocampal-based forms of synaptic plasticity, such as long-term potentiation, are divisible into short- and long-term phases, with the long-term phase requiring the synthesis of new proteins and mRNA for its persistence. By contrast, it is less clear whether long-term depression (LTD) can be divisible into phases. We here describe that in stable hippocampal organotypic cultures, LTD also is not a unitary event but a multiphas...

  14. Protein Degradation and Protection Observed in the Presence of Novel Wound Dressing Components

    OpenAIRE

    Parnell, Laura K.S.

    2011-01-01

    Chronic wounds typically have excessive levels of matrix metalloproteinases (MMPs) and proinflammatory cytokines that impair healing. Reducing these detrimental proteins may be key to healing chronic wounds. Proprietary protease blends were formulated specifically to degrade excessive amounts of proinflammatory factors that could prevent wound healing. Applications of protease-containing wound dressings to acute and chronic wounds have been observed clinically to resolve inflammation and appe...

  15. Blood serum components and serum protein test of Hybro-PG broilers of different ages

    Directory of Open Access Journals (Sweden)

    PRL Silva

    2007-12-01

    Full Text Available Blood serum samples of HYBRO PG broilers were analyzed, with 30 samples collected from 21-day-old broilers (G1, 30 from 35-day-old birds (G2, and 30 from 42-day-old birds (G3, with the aim of establishing normal values of some blood serum parameters. The activities of the enzymes gamma-glutamyl-transferase (GGT, aspartate aminotransferase (AST, creatine kinase (CK, alkaline phosphatase (ALP, and lactate dehydrogenase (LDH, serum levels of total calcium, calcium ion, phosphorus, sodium, potassium, magnesium, chlorides, creatinine, uric acid, triglycerides, cholesterol, total protein, albumin, total and indirect and direct bilirubin, and electrophoretic profile of serum proteins in acrylamide (SDS-PAGE and agarose gel were determined. There was no influence of age on total bilirubin and albumin levels. All the other evaluated parameters presented differences in at least one age group. Protein electrophoretic profile also changed as a function of age. The obtained results can be considered as normal for the studied ages, and therefore be used as references for the interpretation of laboratory exams of broilers of this genetic line in the evaluated ages.

  16. Localization and secretory pathways of a 58K-like protein in multi-vesicular bodies in callus of Arabidopsis thaliana

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Multi-vesicular bodies in endocytosis and protoplasts are special cellular structures that are consid-ered to be originated from invagination of plasma membranes. However, the genesis and function of multi-vesicular bodies, the relationship with Golgi bodies and cell walls, and their secretory pathways remain controversial and ambiguous. Using a monoclonal antibody against an animal 58K protein, we have detected, by Western blotting and confocal microscopy, that a 58K-like protein is present in the calli of Arabidopsis thaliana and Hypericum perforatum. The results of immuno-electron microscopy showed that the 58K-like protein was located in the cisternae of Golgi bodies, secretory vesicles, multi-vesicular bodies, cell walls and vacuoles in callus of Arabidopsis thaliana, suggesting that the multi-vesicular bodies may be originated from Golgi bodies and function as a transporter carrying substances synthesized in Golgi bodies to cell walls and vacuoles. It seems that multi-vesicular bodies have a close relationship with the development of the cell wall and vacuole. The possible secretory pathways of multi-vesicular bodies might be in exocytosis, in which multi-vesicular bodies carry sub-stances to the cell wall for its construction, and in endocytosis, in which multi-vesicular bodies carry substances to the vacuole for its development, depending on what they carry and where the materials are transported. We hence propose that there is more than one pathway for the secretion of multi-vesicular bodies. In addition, our results provided a paradigm that a plant molecule, such as the 58k-like protein in callus of Arabidopsis thaliana, can be detected using a cross-reactive monoclonal antibody induced by an animal protein, and illustrate the existence of analog molecules in both animal and plant kingdoms.

  17. Rear shape in 3 dimensions summarized by principal component analysis is a good predictor of body condition score in Holstein dairy cows.

    Science.gov (United States)

    Fischer, A; Luginbühl, T; Delattre, L; Delouard, J M; Faverdin, P

    2015-07-01

    Body condition is an indirect estimation of the level of body reserves, and its variation reflects cumulative variation in energy balance. It interacts with reproductive and health performance, which are important to consider in dairy production but not easy to monitor. The commonly used body condition score (BCS) is time consuming, subjective, and not very sensitive. The aim was therefore to develop and validate a method assessing BCS with 3-dimensional (3D) surfaces of the cow's rear. A camera captured 3D shapes 2 m from the floor in a weigh station at the milking parlor exit. The BCS was scored by 3 experts on the same day as 3D imaging. Four anatomical landmarks had to be identified manually on each 3D surface to define a space centered on the cow's rear. A set of 57 3D surfaces from 56 Holstein dairy cows was selected to cover a large BCS range (from 0.5 to 4.75 on a 0 to 5 scale) to calibrate 3D surfaces on BCS. After performing a principal component analysis on this data set, multiple linear regression was fitted on the coordinates of these surfaces in the principal components' space to assess BCS. The validation was performed on 2 external data sets: one with cows used for calibration, but at a different lactation stage, and one with cows not used for calibration. Additionally, 6 cows were scanned once and their surfaces processed 8 times each for repeatability and then these cows were scanned 8 times each the same day for reproducibility. The selected model showed perfect calibration and a good but weaker validation (root mean square error=0.31 for the data set with cows used for calibration; 0.32 for the data set with cows not used for calibration). Assessing BCS with 3D surfaces was 3 times more repeatable (standard error=0.075 versus 0.210 for BCS) and 2.8 times more reproducible than manually scored BCS (standard error=0.103 versus 0.280 for BCS). The prediction error was similar for both validation data sets, indicating that the method is not less

  18. The protein scaffold of the lipocalin odorant-binding protein is suitable for the design of new biosensors for the detection of explosive components

    International Nuclear Information System (INIS)

    The detection of hazard exposure is a current priority, including the detection of traces of explosive molecules in different environments like luggage storage rooms and public places, and is becoming a major requirement for homeland security. In the present study we carried out a preliminary investigation on the binding capacities of four forms of the lipocalin odorant-binding protein (OBP) for the detection of explosive components such as diphenylamine, dimethyl-phthalate, resorcinol and dinitrotoluene. The experimental results, showing that OBP binds these compounds with affinity constants ranging between 80 nM and 10.6 mM, indicate that this protein can be used as a probe for the realization of a biosensor to sense explosive compounds

  19. Poly (ADP-Ribose) Polymerase 1 Is Required for Protein Localization to Cajal Body

    OpenAIRE

    Kotova, Elena; Jarnik, Michael; Tulin, Alexei V.

    2009-01-01

    Recently, the nuclear protein known as Poly (ADP-ribose) Polymerase1 (PARP1) was shown to play a key role in regulating transcription of a number of genes and controlling the nuclear sub-organelle nucleolus. PARP1 enzyme is known to catalyze the transfer of ADP-ribose to a variety of nuclear proteins. At present, however, while we do know that the main acceptor for pADPr in vivo is PARP1 protein itself, by PARP1 automodification, the significance of PARP1 automodification for in vivo processe...

  20. Perturbations in phosphoinositide metabolism and protein kinase C activity in mouse liver following whole body irradiation

    International Nuclear Information System (INIS)

    The involvement of the signal transduction pathway in mouse liver following whole body irradiation was investigated. Mice were exposed to 60Co gamma rays (3 Gy) and sacrificed after different time intervals. Various elements of phosphatidyl inositol signal transduction pathway were investigated. Alterations could be seen as early as 15 min of irradiation. These changes are reflected in elevation in DAG levels and increased activation of PKC, an enzyme which is involved in tumorigenesis. The chronological appearance of various transducers following whole body irradiation is of significance since these early effects may set the stage for radiation-induced tumorigenesis and hence may be used to manipulate tumor response to radiotherapy. (author)

  1. A Novel Protein, CHRONO, Functions as a Core Component of the Mammalian Circadian Clock

    OpenAIRE

    Kavaklı, İbrahim Halil; Anafi, Ron C.; Lee, Yoo; Sato, Trey K.; Venkataraman, Anand; Ramanathan, Chidambaram; Hughes, Michael E.; Baggs, Julie E.; Growe, Jacqueline; Liu, Andrew C.; Kim, Junhyong; Hogenesch, John B.

    2014-01-01

    Machine Learning Helps Identify CHRONO as a Circadian Clock Component Ron C. Anafi1,2.*, Yool Lee3., Trey K. Sato3., Anand Venkataraman3, Chidambaram Ramanathan4, Ibrahim H. Kavakli5, Michael E. Hughes6, Julie E. Baggs7, Jacqueline Growe1,2, Andrew C. Liu4, Junhyong Kim8, John B. Hogenesch2,3* 1 Division of Sleep Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America, 2 Center for Sleep and Circadian Neurobiology, Univer...

  2. Expression patterns of Wnt signaling component, secreted frizzled-related protein 3 in astrocytoma and glioblastoma

    Science.gov (United States)

    PEĆINA-ŠLAUS, NIVES; KAFKA, ANJA; VAROŠANEC, ANA MARIA; MARKOVIĆ, LEON; KRSNIK, ŽELJKA; NJIRIĆ, NIKO; MRAK, GORAN

    2016-01-01

    Secreted frizzled-related protein 3 (SFRP3) is a member of the family of soluble proteins, which modulate the Wnt signaling cascade. Novel research has identified aberrant expression of SFRPs in different types of cancer. In the present study the expression intensities and localizations of the SFRP3 protein across different histopathological grades of astrocytic brain tumors were investigated by immunohistochemistry, digital scanning and image analysis. The results demonstrated that the differences between expression levels and malignancy grades were statistically significant. Tumors were classified into four malignancy grades according to the World Health Organization guidelines. Moderate (P=0.014) and strong (P=0.028) nuclear expression levels were significantly different in pilocytic (grade I) and diffuse (grade II) astrocytomas demonstrating higher expression values, as compared with anaplastic astrocytoma (grade III) and glioblastoma (grade IV). When the sample was divided into two groups, the moderate and high cytoplasmic expression levels were observed to be significantly higher in glioblastomas than in the group comprising astrocytoma II and III. Furthermore, the results indicated that high grade tumors were associated with lower values of moderate (P=0.002) and strong (P=0.018) nuclear expression in comparison to low grade tumors. Analysis of cytoplasmic staining demonstrated that strong cytoplasmic expression was significantly higher in the astrocytoma III and IV group than in the astrocytoma I and II group (P=0.048). Furthermore, lower grade astrocytomas exhibited reduced membranous SFRP3 staining when compared with higher grade astrocytomas and this difference was statistically significant (P=0.036). The present results demonstrated that SFRP3 protein expression levels were decreased in the nucleus in higher grade astrocytoma (indicating the expected behavior of an antagonist of Wnt signaling), whereas when the SFRP3 was located in the cytoplasm an

  3. Expression patterns of Wnt signaling component, secreted frizzled‑related protein 3 in astrocytoma and glioblastoma.

    Science.gov (United States)

    Pećina-Šlaus, Nives; Kafka, Anja; Varošanec, Ana Maria; Marković, Leon; Krsnik, Željka; Njirić, Niko; Mrak, Goran

    2016-05-01

    Secreted frizzled-related protein 3 (SFRP3) is a member of the family of soluble proteins, which modulate the Wnt signaling cascade. Novel research has identified aberrant expression of SFRPs in different types of cancer. In the present study the expression intensities and localizations of the SFRP3 protein across different histopathological grades of astrocytic brain tumors were investigated by immunohistochemistry, digital scanning and image analysis. The results demonstrated that the differences between expression levels and malignancy grades were statistically significant. Tumors were classified into four malignancy grades according to the World Health Organization guidelines. Moderate (P=0.014) and strong (P=0.028) nuclear expression levels were significantly different in pilocytic (grade I) and diffuse (grade II) astrocytomas demonstrating higher expression values, as compared with anaplastic astrocytoma (grade III) and glioblastoma (grade IV). When the sample was divided into two groups, the moderate and high cytoplasmic expression levels were observed to be significantly higher in glioblastomas than in the group comprising astrocytoma II and III. Furthermore, the results indicated that high grade tumors were associated with lower values of moderate (P=0.002) and strong (P=0.018) nuclear expression in comparison to low grade tumors. Analysis of cytoplasmic staining demonstrated that strong cytoplasmic expression was significantly higher in the astrocytoma III and IV group than in the astrocytoma I and II group (P=0.048). Furthermore, lower grade astrocytomas exhibited reduced membranous SFRP3 staining when compared with higher grade astrocytomas and this difference was statistically significant (P=0.036). The present results demonstrated that SFRP3 protein expression levels were decreased in the nucleus in higher grade astrocytoma (indicating the expected behavior of an antagonist of Wnt signaling), whereas when the SFRP3 was located in the

  4. Bioinformatics and protein modelling of the GS element of Mycobacteriumavium subsp. paratuberculosis (MAP) and GS-encoded proteins as drugtargets and vaccine components

    Institute of Scientific and Technical Information of China (English)

    Joe Sheridan; Tim Bull; Nazira Sumar; Jun Cheng; John Hermon-Taylor

    2000-01-01

    AIM To determine the function and cellular localization of GS-encoded proteins and to assess their potentialas drug targets and vaccine components.METHODS Bioinformatics software was used to predict the function of GS-encoded proteins and theirlocation within MAP. Protein modelling software was used to build protein structures.RESULTS The gene gsa is a truncated glycosyl transferase and probably non-functional. gsbA and gsbBproduce GDP-fucose which is methylated by gsc and acetylated by mpa. gsd is a fucosyl transferase whichattaches fucose to subterminal rhamnose on cell surface glycopeptidolipid. gsa, gsbA and gsbB and gsc arelocated within the cytoplasm. mpa is embedded in the plasma membrane with 10 transmembrane regions anda conspicuous extracellular loop. gsd is lipid-linked and predicted to localize to the microbial cell surface.CONCLUSION GS encodes the biosynthetic machinery to give MAP a surface coat of methylated andacetylated fucose which may contribute to its protease-resistant nature and ability to minimize immunerecognition. The gsbA/gsbB operon and gsd are promising drug targets and gsd is a good candidatecomponent of a new class of anti-MAP vaccines.

  5. Cupulin Is a Zona Pellucida-Like Domain Protein and Major Component of the Cupula from the Inner Ear

    Science.gov (United States)

    Dernedde, Jens; Weise, Christoph; Müller, Eva-Christina; Hagiwara, Akira; Bachmann, Sebastian; Suzuki, Mamoru; Reutter, Werner; Tauber, Rudolf; Scherer, Hans

    2014-01-01

    The extracellular membranes of the inner ear are essential constituents to maintain sensory functions, the cupula for sensing torsional movements of the head, the otoconial membrane for sensing linear movements and accelerations like gravity, and the tectorial membrane in the cochlea for hearing. So far a number of structural proteins have been described, but for the gelatinous cupula precise data are missing. Here, we describe for the first time a major proteinogenic component of the cupula structure with an apparent molecular mass of 45 kDa from salmon. Analyses of respective peptides revealed highly conserved amino-acid sequences with identity to zona pellucida-like domain proteins. Immunohistochemistry studies localized the protein in the ampulla of the inner ear from salmon and according to its anatomical appearance we identified this glycoprotein as Cupulin. Future research on structure and function of zona pellucida-like domain proteins will enhance our knowledge of inner ear diseases, like sudden loss of vestibular function and other disturbances. PMID:25369234

  6. Pre-emptive Quality Control Protects the ER from Protein Overload via the Proximity of ERAD Components and SRP

    Directory of Open Access Journals (Sweden)

    Hisae Kadowaki

    2015-11-01

    Full Text Available Cells possess ER quality control systems to adapt to ER stress and maintain their function. ER-stress-induced pre-emptive quality control (ER pQC selectively degrades ER proteins via translocational attenuation during ER stress. However, the molecular mechanism underlying this process remains unclear. Here, we find that most newly synthesized endogenous transthyretin proteins are rerouted to the cytosol without cleavage of the signal peptide, resulting in proteasomal degradation in hepatocytes during ER stress. Derlin family proteins (Derlins, which are ER-associated degradation components, reroute specific ER proteins, but not ER chaperones, from the translocon to the proteasome through interactions with the signal recognition particle (SRP. Moreover, the cytosolic chaperone Bag6 and the AAA-ATPase p97 contribute to the degradation of ER pQC substrates. These findings demonstrate that Derlins-mediated substrate-specific rerouting and Bag6- and p97-mediated effective degradation contribute to the maintenance of ER homeostasis without the need for translocation.

  7. A Cyclic di-GMP-binding Adaptor Protein Interacts with Histidine Kinase to Regulate Two-component Signaling.

    Science.gov (United States)

    Xu, Linghui; Venkataramani, Prabhadevi; Ding, Yichen; Liu, Yang; Deng, Yinyue; Yong, Grace Lisi; Xin, Lingyi; Ye, Ruijuan; Zhang, Lianhui; Yang, Liang; Liang, Zhao-Xun

    2016-07-29

    The bacterial messenger cyclic di-GMP (c-di-GMP) binds to a diverse range of effectors to exert its biological effect. Despite the fact that free-standing PilZ proteins are by far the most prevalent c-di-GMP effectors known to date, their physiological function and mechanism of action remain largely unknown. Here we report that the free-standing PilZ protein PA2799 from the opportunistic pathogen Pseudomonas aeruginosa interacts directly with the hybrid histidine kinase SagS. We show that PA2799 (named as HapZ: histidine kinase associated PilZ) binds directly to the phosphoreceiver (REC) domain of SagS, and that the SagS-HapZ interaction is further enhanced at elevated c-di-GMP concentration. We demonstrate that binding of HapZ to SagS inhibits the phosphotransfer between SagS and the downstream protein HptB in a c-di-GMP-dependent manner. In accordance with the role of SagS as a motile-sessile switch and biofilm growth factor, we show that HapZ impacts surface attachment and biofilm formation most likely by regulating the expression of a large number of genes. The observations suggest a previously unknown mechanism whereby c-di-GMP mediates two-component signaling through a PilZ adaptor protein. PMID:27231351

  8. The effects of stress-induced blood components on protein synthesis and secretion in isolated rat hepatocytes

    International Nuclear Information System (INIS)

    The effect of stress-induced blood components were examined, specifically adrenaline and noradrenaline, in the presence and absence of rabbit serum or foetal calf serum, on soluble protein synthesis and secretion by isolated hepatocytes maintained in monolayer culture. Rabbit serum and low doses of adrenaline stimulated soluble protein synthesis and secretion whereas foetal calf serum and high doses of noradrenaline were inhibitory. The effect of noradrenaline on soluble protein synthesis and secretion ocurred in the first 12 hours of incubation. The stimulatory effect of adrenaline was still present after 24 hours of incubation. Preloading of the medium with [3H]-leucine i.e. before the addition of sera and/or catecholamines, showed the [3H]-leucine uptake to have occured to a large extent within the first hour of incubation. Noradrenaline supplementation of the medium at two hourly intervals showed no effect on protein synthesis and secretion. The stability of the cetecholamines and the status of the receptors need to be determined for the effective analysis of the results at any point during the incubation. 17 figs., 15 tabs., 83 refs

  9. Growth performance and certain body measurements of ostrich chicks as affected by dietary protein levels during 2–9 weeks of age

    Directory of Open Access Journals (Sweden)

    Kh.M. Mahrose

    2015-07-01

    Full Text Available The present work was conducted to examine the effects of dietary crude protein (CP levels (18, 21 and 24% on growth performance (Initial and final body weight, daily body weight gain, feed consumption, feed conversion and protein efficiency ratio during 2-9 weeks of age and certain body measurements (body height, tibiotarsus length and tibiotarsus girth at 9 weeks of age. A total of 30 African Black unsexed ostrich chicks were used in the present study in simple randomized design. The results of the present work indicated that initial and final live body weight, body weight gain, feed consumption, feed conversion of ostrich chicks were insignificantly affected by dietary protein level used. Protein efficiency ratio was high in the group of chicks fed diet contained 18% CP. Results obtained indicated that tibiotarsus girth was decreased (P≤0.01 with the increasing dietary protein level, where the highest value of tibiotarsus girth (18.38 cm was observed in chicks fed 18% dietary protein level. Body height and tibiotarsus length were not significantly different. In conclusion, the results of the present study indicate that ostrich chicks (during 2-9 weeks of age could grow on diets contain lower levels of CP (18%.

  10. Studies of a nuclear matrix protein restricted to normal brain cells and lead-induced intranuclear inclusion bodies of kidney

    Energy Technology Data Exchange (ETDEWEB)

    Shelton, K.; Egle, P.; Redford, K.; Bigbee, J.

    1986-05-01

    A nuclear matrix protein, p32/6.3, with an unusual tissue distribution, has been identified. Protein from 21 tissues was surveyed by immunoprobing Western blots. In normal adult rats p32/6.3 is found only in grey matter from the cerebrum and the cerebellum, occurring in both neurons and astrocytes. Other brain cell types have not been examined. The protein appears to be developmentally regulated. It is detectable in the brain within a few days after birth and reaches adult levels within one to two weeks. Brain p32/6.3 has been found in all animals tested including rat, mouse, dog, cow, pig, chicken and human. This conservation indicates a fundamental role for p32/6.3 in the nucleus of brain cells. Possible functions for p32/6.3 may be indicated by a second novel occurrence. Chronic lead poisoning characteristically induces intranuclear inclusion bodies in the cells lining kidney proximal tubules. p32/6.3 is a major constituent of these inclusion bodies. They are also rich in lead and other metals including calcium, iron, zinc, copper and cadmium. These diverse observations suggest that p32/6.3 may have a role in metal homeostasis in the brain of normal animals.

  11. Effects of a high protein diet on body weight and comorbidities associated with obesity.

    Science.gov (United States)

    Clifton, Peter

    2012-08-01

    Red meat intake has been frequently associated with the development of coronary artery disease and type 2 diabetes but vegetable protein has been associated with protection from these diseases. Whether this is related to the protein per se or to the increased polyunsaturated fat or higher fibre levels associated with more vegetarian diets is not clear. High protein diets are associated with greater satiety and in some studies are associated with greater weight loss compared with high carbohydrate diets especially in an ad libitum design. These diets also lower plasma triglyceride and blood pressure and sometimes spare lean mass. There appear to be no harmful effects of high protein diets on bone density or renal function in weight loss studies. PMID:23107523

  12. Increased Body Mass Index, Elevated C-reactive Protein, and Short Telomere Length

    DEFF Research Database (Denmark)

    Rode, Line; Nordestgaard, Børge G; Weischer, Maren; Bojesen, Stig E

    2014-01-01

    protein. SETTING AND DESIGN: We studied 45,069 individuals from the Copenhagen General Population Study with measurements of leukocyte telomere length, BMI, and C-reactive protein in a Mendelian randomization study. Using the three obesity-associated polymorphisms FTO rs9939609, MC4R rs17782313, and TMEM......18 rs6548238, and the CRP promoter polymorphism rs3091244 in instrumental variable analyses, we estimated the associations between genetically increased BMI and telomere length and between genetically increased C-reactive protein and telomere length. RESULTS: In multivariable-adjusted observational...... analyses, telomere length decreased with seven base pairs (95% confidence interval, -9--5) per unit increase in BMI, and further adjustment for C-reactive protein attenuated this association to -5 base pairs (-8--3). In accordance, instrumental variable analysis showed a non-significant telomere length...

  13. Norvaline and Norleucine May Have Been More Abundant Protein Components during Early Stages of Cell Evolution

    Science.gov (United States)

    Alvarez-Carreño, Claudia; Becerra, Arturo; Lazcano, Antonio

    2013-10-01

    The absence of the hydrophobic norvaline and norleucine in the inventory of protein amino acids is readdressed. The well-documented intracellular accumulation of these two amino acids results from the low-substrate specificity of the branched-chain amino acid biosynthetic enzymes that act over a number of related α-ketoacids. The lack of absolute substrate specificity of leucyl-tRNA synthase leads to a mischarged norvalyl-tRNALeu that evades the translational proofreading activites and produces norvaline-containing proteins, (cf. Apostol et al. J Biol Chem 272:28980-28988, 1997). A similar situation explains the presence of minute but detectable amounts of norleucine in place of methionine. Since with few exceptions both leucine and methionine are rarely found in the catalytic sites of most enzymes, their substitution by norvaline and norleucine, respectively, would have not been strongly hindered in small structurally simple catalytic polypeptides during the early stages of biological evolution. The report that down-shifts of free oxygen lead to high levels of intracellular accumulation of pyruvate and the subsequent biosynthesis of norvaline (Soini et al. Microb Cell Factories 7:30, 2008) demonstrates the biochemical and metabolic consequences of the development of a highly oxidizing environment. The results discussed here also suggest that a broader definition of biomarkers in the search for extraterrestrial life may be required.

  14. Immunolocalization of von Willebrand protein in Weibel-Palade bodies of human endothelial cells

    OpenAIRE

    1982-01-01

    Immunofluorescence staining of cultured human umbilical vein endothelial cells has shown the presence of von Willebrand protein in the perinuclear region, in small rodlike structures through the cytoplasm, and on filaments of the extracellular matrix. Nonendothelial cells showed no staining with anti-von Willebrand protein antiserum. At the light microscope level, immunoperoxidase treatment of endothelial cells revealed the same pattern and antibody specificity as the fluorescence staining. T...

  15. Characterization of a nuclear compartment shared by nuclear bodies applying ectopic protein expression and correlative light and electron microscopy

    International Nuclear Information System (INIS)

    To investigate the accessibility of interphase nuclei for nuclear body-sized particles, we analyzed in cultured cells from human origin by correlative fluorescence and electron microscopy (EM) the bundle-formation of Xenopus-vimentin targeted to the nucleus via a nuclear localization signal (NLS). Moreover, we investigated the spatial relationship of speckles, Cajal bodies, and crystalline particles formed by Mx1 fused to yellow fluorescent protein (YFP), with respect to these bundle arrays. At 37 deg C, the nucleus-targeted, temperature-sensitive Xenopus vimentin was deposited in focal accumulations. Upon shift to 28 deg C, polymerization was induced and filament arrays became visible. Within 2 h after temperature shift, arrays were found to be composed of filaments loosely embedded in the nucleoplasm. The filaments were restricted to limited areas of the nucleus between focal accumulations. Upon incubation at 28 deg C for several hours, NLS vimentin filaments formed bundles looping throughout the nuclei. Speckles and Cajal bodies frequently localized in direct neighborhood to vimentin bundles. Similarly, small crystalline particles formed by YFP-tagged Mx1 also located next to vimentin bundles. Taking into account that nuclear targeted vimentin locates in the interchromosomal domain (ICD), we conclude that nuclear body-sized particles share a common nuclear space which is controlled by higher order chromatin organization

  16. Intermittent fasting does not affect whole-body glucose, lipid, or protein metabolism

    NARCIS (Netherlands)

    M.R. Soeters; N.M. Lammers; P.F. Dubbelhuis; M.T. Ackermans; C.F. Jonkers-Schuitema; E. Fliers; H.P. Sauerwein; J.M. Aerts; M.J. Serlie

    2009-01-01

    Background: Intermittent fasting (IF) was shown to increase whole-body insulin sensitivity, but it is uncertain whether IF selectively influences intermediary metabolism. Such selectivity might be advantageous when adapting to periods of food abundance and food shortage. Objective: The objective was

  17. Sorting of proteins into multivesicular bodies : ubiquitin-dependent and -independent targeting

    NARCIS (Netherlands)

    Reggiori, F; Pelham, H R; Reggiori, Fulvio

    2001-01-01

    Yeast endosomes, like those in animal cells, invaginate their membranes to form internal vesicles. The resulting multivesicular bodies fuse with the vacuole, the lysosome equivalent, delivering the internal vesicles for degradation. We have partially purified internal vesicles and analysed their con

  18. An essential nuclear protein in trypanosomes is a component of mRNA transcription/export pathway.

    Directory of Open Access Journals (Sweden)

    Mariana Serpeloni

    Full Text Available In eukaryotic cells, different RNA species are exported from the nucleus via specialized pathways. The mRNA export machinery is highly integrated with mRNA processing, and includes a different set of nuclear transport adaptors as well as other mRNA binding proteins, RNA helicases, and NPC-associated proteins. The protozoan parasite Trypanosoma cruzi is the causative agent of Chagas disease, a widespread and neglected human disease which is endemic to Latin America. Gene expression in Trypanosoma has unique characteristics, such as constitutive polycistronic transcription of protein-encoding genes and mRNA processing by trans-splicing. In general, post-transcriptional events are the major points for regulation of gene expression in these parasites. However, the export pathway of mRNA from the nucleus is poorly understood. The present study investigated the function of TcSub2, which is a highly conserved protein ortholog to Sub2/ UAP56, a component of the Transcription/Export (TREX multiprotein complex connecting transcription with mRNA export in yeast/human. Similar to its orthologs, TcSub2 is a nuclear protein, localized in dispersed foci all over the nuclei -except the fibrillar center of nucleolus- and at the interface between dense and non-dense chromatin areas, proposing the association of TcSub2 with transcription/processing sites. These findings were analyzed further by BrUTP incorporation assays and confirmed that TcSub2 is physically associated with active RNA polymerase II (RNA pol II, but not RNA polymerase I (RNA pol I or Spliced Leader (SL transcription, demonstrating participation particularly in nuclear mRNA metabolism in T. cruzi. The double knockout of the TcSub2 gene is lethal in T. cruzi, suggesting it has an essential function. Alternatively, RNA interference assays were performed in Trypanosoma brucei. It allowed demonstrating that besides being an essential protein, its knockdown causes mRNA accumulation in the nucleus and

  19. The Significance of Protein Components in Heterogeneous Eggs for Embrionic Development and Larvas Persistence of Cyprinidae

    Directory of Open Access Journals (Sweden)

    Zalepukhin Valeriy Vladimirovich

    2014-12-01

    Full Text Available Endogenous heterogeneity of females and their eggs is one of the most important factors of annual generation’s formation in nature-spawn and incubative centers. Its assessment is significant for comprehending the notion of biochemical changes in proximate prespawn period. Between the shares of egg’s biochemical composition of Cyprinidae species only protein and b-lipoproteids levels are incovering constant and authentic correlations with embrionic development and larval survival in the conditions of artificial reproduction. The promotion of protein’s level to 20 – 24 % (in wet weigth in percolating eggs positively affects the fertilization and larvae vitality. This trend is fair for the domesticated and nature-spawn fish. The same significance is important for the b-lipoproteids level in ovular eggs of grass carp (Ctenopharyngodon idella Val.. The optimal concentration is 100-200 mg % in wet weight.

  20. [Effects of water-nitrogen interaction on the contents and components of protein and starch in wheat grains].

    Science.gov (United States)

    Fu, Xue-Li; Wang, Chen-Yang; Guo, Tian-Cai; Zhu, Yun-Ji; Ma, Dong-Yun; Wang, Yong-Hua

    2008-02-01

    With wheat cultivars Yumai 34 (strong-gluten wheat) and Yumai 50 (weak-gluten wheat) as test materials, a field experiment was conducted to study the effects of three irrigation treatments (irrigation at jointing stage, at jointing and grain-filling stages, and at jointing, grain-filling, and pre-maturing stages), three nitrogen application rates (0, 150, and 270 kg x hm(-2)), and their combinations on the contents and components of protein and starch in wheat grains. The results showed that for strong-gluten wheat cultivar Yumai 34, applying 270 kg x hm(-2) of N increased the total content of protein and the contents of albumin, gliadin and glutelin, and enhanced the glutelin/gliadin ratio. This application rate of nitrogen also increased the total content of starch and the content of amylopectin, and decreased the amylose/amylopetin ratio. For weak-gluten wheat cultivar Yumai 50, applying 150 kg x hm(-2) of N increased the contents of albumin and gliadin, and decreased the contents of globulin and glutelin and the glutelin/gliadin ratio. The amylopectin and starch contents also increased when the N application rate was 150 kg x hm(-2). Non-N fertilization or applying 270 kg x hm(-2) of N decreased the accumulation of protein and starch, and resulted in a decrease of grain yield. Among the irrigation treatments, irrigation at jointing and grain-filling stages promoted the accumulation of protein and starch in grains and increased the grain yield, while the other two treatments were unbeneficial to the accumulation of protein and starch and decreased the grain yield. Applying 270 kg x hm(-2) and 150 kg x hm(-2) of N combined with irrigation at jointing and grain-filling stages was the ideal management regime for the high yield and good quality of strong- and weak-gluten wheat cultivars, respectively. PMID:18464637

  1. Detailed NMR Analysis of the Heme-Protein Interactions in Component IV Glycera dibranchiata Monomeric Hemoglobin-CO

    International Nuclear Information System (INIS)

    Complete 13C, 15N, and 1H resonance assignments have been obtained for the recombinant, ferrous CO-ligated form of component IV monomeric hemoglobin from Glycera dibranchiata. This 15642 Da myoglobin-like protein contains a large number of glycine and alanine residues (47) and a heme prosthetic group. Coupling constant information has allowed the determination of χ1 and χ2 torsion angles, backbone φ angles, as well as 43 of 81 possible assignments to Hβ2/β3 pairs. The 13Cα, 13Cβ, 13C', and 1Hα assignments yield a consensus chemical shift index (CSI) that, in combination with NOE information and backbone torsion angles, defines seven distinct helical regions for the protein's global architecture. Discrepancies between the CSI and NOE/3JHNHα-based secondary structure definitions have been attributed to heme ring current shifts on the basis of calculations from a model structure [Alam et al. (1994) J. Protein Chem., 13, 151-164]. The agreement can be improved by correcting the 1Hα chemical shifts for the ring current contributions. Because the holoprotein was assembled from isotopically enriched globin and natural isotope-abundance heme, data from 13C-filtered/13C-edited and 13C-filtered/13C-filtered 2D NOESY experiments could be used to determine complete heme proton assignments and to position the heme within the protein. The results confirm the unusual presence of Phe31(B10) and Leu58(E7) side chains near the heme ligand binding site which may alter the polarity and steric environment and thus the functional properties of this protein

  2. Functional analysis of Abp1p-interacting proteins involved in endocytosis of the MCC component in Aspergillus oryzae.

    Science.gov (United States)

    Matsuo, Kento; Higuchi, Yujiro; Kikuma, Takashi; Arioka, Manabu; Kitamoto, Katsuhiko

    2013-07-01

    We have investigated the functions of three endocytosis-related proteins in the filamentous fungus Aspergillus oryzae. Yeast two-hybrid screening using the endocytic marker protein AoAbp1 (A.oryzae homolog of Saccharomyces cerevisiae Abp1p) as a bait identified four interacting proteins named Aip (AoAbp1 interacting proteins). In mature hyphae, EGFP (enhanced green fluorescent protein) fused to Aips colocalized with AoAbp1 at the hyphal tip region and the plasma membrane, suggesting that Aips function in endocytosis. aipA is a putative AAA ATPase and its function has been dissected (Higuchi et al., 2011). aipB, the homolog of A. nidulans myoA, encodes an essential class I myosin and its conditional mutant showed a germination defect. aipC and aipD do not contain any recognizable domains except some proline-rich regions which may interact with two SH3 (Src homology 3) domains of AoAbp1. Neither aipC nor aipD disruptants showed any defects in their growth, but the aipC disruptant formed less conidia compared with the control strain. In addition, the aipC disruptant was resistant to the triazole antifungal drugs that inhibit ergosterol biosynthesis. Although no aip disruptants showed any defects in the uptake of the fluorescent dye FM4-64, the endocytosis of the arginine permease AoCan1, one of the MCC (membrane compartment of Can1p) components, was delayed in both aipC and aipD disruptants. In A. oryzae, AoCan1 localized mainly at the plasma membrane in the basal region of hyphae, suggesting that different endocytic mechanisms exist in apical and basal regions of highly polarized cells. PMID:23597630

  3. SSX2 is a novel DNA-binding protein that antagonizes polycomb group body formation and gene repression

    DEFF Research Database (Denmark)

    Gjerstorff, Morten; Greve, Katrine Buch Vidén; Møller, Jesper Bonnet; Elias, Daniel; Schmidt, Steffen; Mollenhauer, Jan; Volborg, Bjørn; Møllegaard, NIels Erik; Ditzel, Henrik

    Polycomb group (PcG) complexes regulate cellular identity through epigenetic programming of chromatin. Here, we show that SSX2, a germline-specific protein ectopically expressed in melanoma, is a chromatin-associated protein that antagonizes BMI1 and EZH2 PcG body formation and derepresses Pc......G target genes. SSX2 further negatively regulates the level of the PcG-associated histone mark H3K27me3 in melanoma cells, and there is a clear inverse correlation between SSX2/3 expression and H3K27me3 in spermatogenesis. SSX2 does not affect the overall composition and stability of PcG complexes, but SSX...

  4. Isolation of soybean protein P34 from oil bodies using hydrophobic interaction chromatography

    OpenAIRE

    Seidel-Morgenstern Andreas; Kessler Lars; Sewekow Eva; Rothkötter Hermann-Josef

    2008-01-01

    Background Soybeans play a prominent role in allergologic research due to the high incidence of allergic reactions. For detailed studies on specific proteins it is necessary to have access to a large amount of pure substance. Results In this contribution, a method for purifying soybean (Glycine max) protein P34 (also called Gly m Bd 30K or Gly m 1) using hydrophobic interaction chromatography is presented. After screening experiments using 1 mL HiTrap columns, Butyl Sepharose 4 FF w...

  5. Contact- and distance-based principal component analysis of protein dynamics

    International Nuclear Information System (INIS)

    To interpret molecular dynamics simulations of complex systems, systematic dimensionality reduction methods such as principal component analysis (PCA) represent a well-established and popular approach. Apart from Cartesian coordinates, internal coordinates, e.g., backbone dihedral angles or various kinds of distances, may be used as input data in a PCA. Adopting two well-known model problems, folding of villin headpiece and the functional dynamics of BPTI, a systematic study of PCA using distance-based measures is presented which employs distances between Cα-atoms as well as distances between inter-residue contacts including side chains. While this approach seems prohibitive for larger systems due to the quadratic scaling of the number of distances with the size of the molecule, it is shown that it is sufficient (and sometimes even better) to include only relatively few selected distances in the analysis. The quality of the PCA is assessed by considering the resolution of the resulting free energy landscape (to identify metastable conformational states and barriers) and the decay behavior of the corresponding autocorrelation functions (to test the time scale separation of the PCA). By comparing results obtained with distance-based, dihedral angle, and Cartesian coordinates, the study shows that the choice of input variables may drastically influence the outcome of a PCA

  6. A novel protein, CHRONO, functions as a core component of the mammalian circadian clock.

    Directory of Open Access Journals (Sweden)

    Akihiro Goriki

    2014-04-01

    Full Text Available Circadian rhythms are controlled by a system of negative and positive genetic feedback loops composed of clock genes. Although many genes have been implicated in these feedback loops, it is unclear whether our current list of clock genes is exhaustive. We have recently identified Chrono as a robustly cycling transcript through genome-wide profiling of BMAL1 binding on the E-box. Here, we explore the role of Chrono in cellular timekeeping. Remarkably, endogenous CHRONO occupancy around E-boxes shows a circadian oscillation antiphasic to BMAL1. Overexpression of Chrono leads to suppression of BMAL1-CLOCK activity in a histone deacetylase (HDAC -dependent manner. In vivo loss-of-function studies of Chrono including Avp neuron-specific knockout (KO mice display a longer circadian period of locomotor activity. Chrono KO also alters the expression of core clock genes and impairs the response of the circadian clock to stress. CHRONO forms a complex with the glucocorticoid receptor and mediates glucocorticoid response. Our comprehensive study spotlights a previously unrecognized clock component of an unsuspected negative circadian feedback loop that is independent of another negative regulator, Cry2, and that integrates behavioral stress and epigenetic control for efficient metabolic integration of the clock.

  7. Contact- and distance-based principal component analysis of protein dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Ernst, Matthias; Sittel, Florian; Stock, Gerhard, E-mail: stock@physik.uni-freiburg.de [Biomolecular Dynamics, Institute of Physics, Albert Ludwigs University, 79104 Freiburg (Germany)

    2015-12-28

    To interpret molecular dynamics simulations of complex systems, systematic dimensionality reduction methods such as principal component analysis (PCA) represent a well-established and popular approach. Apart from Cartesian coordinates, internal coordinates, e.g., backbone dihedral angles or various kinds of distances, may be used as input data in a PCA. Adopting two well-known model problems, folding of villin headpiece and the functional dynamics of BPTI, a systematic study of PCA using distance-based measures is presented which employs distances between C{sub α}-atoms as well as distances between inter-residue contacts including side chains. While this approach seems prohibitive for larger systems due to the quadratic scaling of the number of distances with the size of the molecule, it is shown that it is sufficient (and sometimes even better) to include only relatively few selected distances in the analysis. The quality of the PCA is assessed by considering the resolution of the resulting free energy landscape (to identify metastable conformational states and barriers) and the decay behavior of the corresponding autocorrelation functions (to test the time scale separation of the PCA). By comparing results obtained with distance-based, dihedral angle, and Cartesian coordinates, the study shows that the choice of input variables may drastically influence the outcome of a PCA.

  8. Ovulation-inducing factor: a protein component of llama seminal plasma

    Directory of Open Access Journals (Sweden)

    Huanca Wilfredo

    2010-05-01

    Full Text Available Abstract Background Previously, we documented the presence of ovulation-inducing factor (OIF in the seminal plasma of llamas and alpacas. The purpose of the study was to define the biochemical characteristics of the molecule(s in seminal plasma responsible for inducing ovulation. Methods In Experiment 1, llama seminal plasma was centrifuged using filtration devices with nominal molecular mass cut-offs of 30, 10 and 5 kDa. Female llamas (n = 9 per group were treated i.m. with whole seminal plasma (positive control, phosphate-buffered saline (negative control, or the fraction of seminal plasma equal or higher than 30 kDa, 10 to 30 kDa, 5 to 10 kDa, or Results In Experiment 1, all llamas in the equal or higher than 30 kDa and positive control groups ovulated (9/9 in each, but none ovulated in the other groups (P Conclusions We conclude that ovulation-inducing factor (OIF in llama seminal plasma is a protein molecule that is resistant to heat and enzymatic digestion with proteinase K, and has a molecular mass of approximately equal or higher than 30 kDa.

  9. Modulation of function of multidrug resistance associated-proteins by Kaempferia parviflora extracts and their components.

    Science.gov (United States)

    Patanasethanont, Denpong; Nagai, Junya; Matsuura, Chie; Fukui, Kyoko; Sutthanut, Khaetthareeya; Sripanidkulchai, Bung-orn; Yumoto, Ryoko; Takano, Mikihisa

    2007-07-01

    In this study, the effects of extracts and flavone derivatives from the rhizome of Kaempferia parviflora on multidrug resistance associated-proteins (MRP)-mediated transport in A549 cells were examined. The cells employed express MRP1 and MRP2, but not P-glycoprotein. The cellular accumulation of calcein, an MRP substrate, was significantly increased by various MRP inhibitors without being affected by verapamil, a typical P-glycoprotein inhibitor. Ethanol and aqueous extracts from K. parviflora rhizome increased the accumulation of calcein and doxorubicin in A549 cells in a concentration-dependent manner. The inhibitory potency of the ethanol extract for MRP function was greater than that of the aqueous extract. Among six flavone derivatives isolated from K. parviflora rhizome, 5,7-dimethoxyflavone exhibited a maximal stimulatory effect on the accumulation of doxorubicin in A549 cells. The accumulation of doxorubicin was increased by four flavone derivatives without 5-hydroxy group, but not by the other two flavone derivatives with 5-hydroxy group. In addition, 5,7-dimethoxyflavone and 3,5,7,3',4'-pentamethoxyflavone decreased resistance to doxorubicin in A549 cells. These findings indicate that extracts and flavone derivatives from the rhizome of K. parviflora suppress MRP function, and therefore may be useful as modulators of multidrug resistance in cancer cells. PMID:17481606

  10. A Model of Yeast Cell-Cycle Regulation Based on a Standard Component Modeling Strategy for Protein Regulatory Networks.

    Directory of Open Access Journals (Sweden)

    Teeraphan Laomettachit

    Full Text Available To understand the molecular mechanisms that regulate cell cycle progression in eukaryotes, a variety of mathematical modeling approaches have been employed, ranging from Boolean networks and differential equations to stochastic simulations. Each approach has its own characteristic strengths and weaknesses. In this paper, we propose a "standard component" modeling strategy that combines advantageous features of Boolean networks, differential equations and stochastic simulations in a framework that acknowledges the typical sorts of reactions found in protein regulatory networks. Applying this strategy to a comprehensive mechanism of the budding yeast cell cycle, we illustrate the potential value of standard component modeling. The deterministic version of our model reproduces the phenotypic properties of wild-type cells and of 125 mutant strains. The stochastic version of our model reproduces the cell-to-cell variability of wild-type cells and the partial viability of the CLB2-dbΔ clb5Δ mutant strain. Our simulations show that mathematical modeling with "standard components" can capture in quantitative detail many essential properties of cell cycle control in budding yeast.

  11. A Model of Yeast Cell-Cycle Regulation Based on a Standard Component Modeling Strategy for Protein Regulatory Networks.

    Science.gov (United States)

    Laomettachit, Teeraphan; Chen, Katherine C; Baumann, William T; Tyson, John J

    2016-01-01

    To understand the molecular mechanisms that regulate cell cycle progression in eukaryotes, a variety of mathematical modeling approaches have been employed, ranging from Boolean networks and differential equations to stochastic simulations. Each approach has its own characteristic strengths and weaknesses. In this paper, we propose a "standard component" modeling strategy that combines advantageous features of Boolean networks, differential equations and stochastic simulations in a framework that acknowledges the typical sorts of reactions found in protein regulatory networks. Applying this strategy to a comprehensive mechanism of the budding yeast cell cycle, we illustrate the potential value of standard component modeling. The deterministic version of our model reproduces the phenotypic properties of wild-type cells and of 125 mutant strains. The stochastic version of our model reproduces the cell-to-cell variability of wild-type cells and the partial viability of the CLB2-dbΔ clb5Δ mutant strain. Our simulations show that mathematical modeling with "standard components" can capture in quantitative detail many essential properties of cell cycle control in budding yeast. PMID:27187804

  12. AlgK is a TPR-containing Protein and the Periplasmic Component of a Novel Exopolysaccharide Secretin

    Energy Technology Data Exchange (ETDEWEB)

    Keiski, C.; Harwich, M; Jain, S; Neculai, A; Whitney, J; Yip, P; Robinson, H; Riley, L; Burrows, L; et al.

    2010-01-01

    The opportunistic pathogen Pseudomonas aeruginosa causes chronic biofilm infections in cystic fibrosis patients. During colonization of the lung, P. aeruginosa converts to a mucoid phenotype characterized by overproduction of the exopolysaccharide alginate. Here we show that AlgK, a protein essential for production of high molecular weight alginate, is an outer membrane lipoprotein that contributes to the correct localization of the porin AlgE. Our 2.5 {angstrom} structure shows AlgK is composed of 9.5 tetratricopeptide-like repeats, and three putative sites of protein-protein interaction have been identified. Bioinformatics analysis suggests that BcsA, PgaA, and PelB, involved in the production and export of cellulose, poly-{beta}-1,6-N-Acetyl-d-glucosamine, and Pel exopolysaccharide, respectively, share the same topology as AlgK/E. Together, our data suggest that AlgK plays a role in the assembly of the alginate biosynthetic complex and represents the periplasmic component of a new type of outer membrane secretin that differs from canonical bacterial capsular polysaccharide secretion systems.

  13. Sequence and Function of LuxU: a Two-Component Phosphorelay Protein That Regulates Quorum Sensing in Vibrio harveyi

    Science.gov (United States)

    Freeman, Jeremy A.; Bassler, Bonnie L.

    1999-01-01

    Vibrio harveyi regulates the expression of bioluminescence (lux) in response to cell density, a phenomenon known as quorum sensing. In V. harveyi, two independent quorum-sensing systems exist, and each produces, detects, and responds to a specific cell density-dependent autoinducer signal. The autoinducers are recognized by two-component hybrid sensor kinases called LuxN and LuxQ, and sensory information from both systems is transduced by a phosphorelay mechanism to the response regulator protein LuxO. Genetic evidence suggests that LuxO-phosphate negatively regulates the expression of luminescence at low cell density in the absence of autoinducers. At high cell density, interaction of the sensors with their cognate autoinducers results in dephosphorylation and inactivation of the LuxO repressor. In the present report, we show that LuxN and LuxQ channel sensory information to LuxO via a newly identified phosphorelay protein that we have named LuxU. LuxU shows sequence similarity to other described phosphorelay proteins, including BvgS, ArcB, and Ypd1. A critical His residue (His 58) of LuxU is required for phosphorelay function. PMID:9922254

  14. Health issues of whey proteins: 1. Protection of lean body mass

    NARCIS (Netherlands)

    Schaafsma, G.

    2006-01-01

    Loss of muscle mass as a consequence of changes in protein metabolism during periods of catabolic stress is a serious complication in a variety of conditions. These conditions are weight loss programs, sarcopenia in the elderly and several clinical states. It appears from many studies that improved

  15. Association between dietary protein and change in body composition among children (EYHS)

    DEFF Research Database (Denmark)

    van Vught, Anneke J A H; Heitmann, Berit L; Nieuwenhuizen, Arie G;

    2009-01-01

    mass index (FFMI) and fat mass index (FMI), based on skinfold measurements. Dietary intake was estimated via 24h recall. Associations between intakes of protein as well as arginine, lysine and change in FFMI and FMI were analysed by multiple linear regressions, adjusted for social economic status...

  16. Contribution of individual components of a job cycle on overall severity of whole-body vibration exposure: a study in Indian mines.

    Science.gov (United States)

    Mandal, Bibhuti B; Mansfield, Neil J

    2016-01-01

    Drivers of earth-moving machines are exposed to whole-body vibration (WBV). In mining operations there can be a combination of relatively high magnitudes of vibration and long exposure times. Effective risk mitigation requires understanding of the main aspects of a task that pose a hazard to health. There are very few published studies of WBV exposure from India. This paper reports on a study that considered the contribution of the component phases of dumper operations, on the overall vibration exposure of the drivers. It shows that vibration magnitudes are relatively high, and that haulage tasks are the main contributor to the exposure. It is recommended that driver speed, haul road surfaces and vehicle maintenance/selection are optimized to ensure minimization of vibration. If this is not sufficient, operation times might need to be reduced in order to ensure that the health guidance caution zone from Standard No. ISO 2631-1:1997 is not exceeded. PMID:26652833

  17. The Chloroplast Protein Translocation Complexes of Chlamydomonas reinhardtii: A Bioinformatic Comparison of Toc and Tic Components in Plants, Green Algae and Red Algae

    OpenAIRE

    Kalanon, Ming; McFadden, Geoffrey I

    2008-01-01

    The recently completed genome of Chlamydomonas reinhardtii was surveyed for components of the chloroplast protein translocation complexes. Putative components were identified using reciprocal BlastP searches with the protein sequences of Arabidopsis thaliana as queries. As a comparison, we also surveyed the new genomes of the bryophyte Physcomitrella patens, two prasinophyte green algae (Ostreococcus lucimarinus and Ostreococcus tauri), the red alga Cyanidioschizon merolae, and several cyanob...

  18. The relationship of high sensitivity C-reactive protein to percent body fat mass, body mass index, waist-to-hip ratio, and waist circumference in a Taiwanese population

    Directory of Open Access Journals (Sweden)

    Lin Wen-Yuan

    2010-09-01

    Full Text Available Abstract Background High-sensitivity C-reactive protein (hs-CRP is an easily measured inflammatory biomarker. This study compared the association of percent body fat mass (%FM, body mass index (BMI, waist circumference (WC, and waist-to-hip ratio (WHR with hs-CRP in a Taiwanese population. Methods A total of 1669 subjects aged 40-88 years were recruited in 2004 in a metropolitan city in Taiwan. The relationships between obesity indicators and a high level of hs-CRP were examined using multivariate logistic regression analysis. The upper quartile of the hs-CRP distributions was defined as the high category group. The areas under the curve (AUCs of the receiver operating characteristic curves were calculated for all obesity indicators to compare their relative ability to correctly classify subjects with a high level of hs-CRP. Results After multivariate adjustment, the odds ratio for %FM was the only significant indicator that was associated with a high level of hs-CRP in men (1.55, 95% CI: 1.07-2.25. All indicators were associated with a high level of hs-CRP in women. In men, the AUCs for %FM were significantly higher than those for BMI, WHR, and WC, when demographic and lifestyle behaviors were considered (p Conclusions Our study demonstrates that %FM is the only obesity indicator that is strongly associated with a high level of hs-CRP after adjusting for sociodemographic factors, lifestyle behaviors and components of metabolic syndrome in both genders in a Taiwanese population aged forty years and over. In men, %FM had the greatest ability to classify subjects with a high level of hs-CRP when only demographic and lifestyle behaviors were considered. Our study finding has important implications for the screening of obesity in community settings.

  19. Minimized human telomerase maintains telomeres and resolves endogenous roles of H/ACA proteins, TCAB1, and Cajal bodies.

    Science.gov (United States)

    Vogan, Jacob M; Zhang, Xiaozhu; Youmans, Daniel T; Regalado, Samuel G; Johnson, Joshua Z; Hockemeyer, Dirk; Collins, Kathleen

    2016-01-01

    We dissected the importance of human telomerase biogenesis and trafficking pathways for telomere maintenance. Biological stability of human telomerase RNA (hTR) relies on H/ACA proteins, but other eukaryotes use other RNP assembly pathways. To investigate additional rationale for human telomerase assembly as H/ACA RNP, we developed a minimized cellular hTR. Remarkably, with only binding sites for telomerase reverse transcriptase (TERT), minimized hTR assembled biologically active enzyme. TERT overexpression was required for cellular interaction with minimized hTR, indicating that H/ACA RNP assembly enhances endogenous hTR-TERT interaction. Telomere maintenance by minimized telomerase was unaffected by the elimination of the telomerase holoenzyme Cajal body chaperone TCAB1 or the Cajal body scaffold protein Coilin. Surprisingly, wild-type hTR also maintained and elongated telomeres in TCAB1 or Coilin knockout cells, with distinct changes in telomerase action. Overall, we elucidate trafficking requirements for telomerase biogenesis and function and expand mechanisms by which altered telomere maintenance engenders human disease. PMID:27525486

  20. The Arrestin Domain Containing 3 (ARRDC3) Protein Regulates Body Mass and Energy Expenditure

    OpenAIRE

    Patwari, Parth; Emilsson, Valur; Schadt, Eric E.; Chutkow, William A.; Lee, Samuel; Marsili, Alessandro; Zhang, Yongzhao; Dobrin, Radu; Cohen, David E.; Larsen, P. Reed; Zavacki, Ann Marie; Fong, Loren G; Young, Stephen G.; Lee, Richard T.

    2011-01-01

    A human genome-wide linkage scan for obesity identified a linkage peak on chromosome 5q13–15. Positional cloning revealed an association of a rare haplotype to high body-mass index (BMI) in males but not females. The risk locus contains a single gene, “arrestin domain containing 3” (ARRDC3), an uncharacterized α-arrestin. Inactivating Arrdc3 in mice led to a striking resistance to obesity, with greater impact on male mice. Mice with decreased ARRDC3 levels were protected from obesity due to i...

  1. The classical Ubisch bodies carry a sporophytically produced structural protein (RAFTIN) that is essential for pollen development.

    Science.gov (United States)

    Wang, Aiming; Xia, Qun; Xie, Wenshuang; Datla, Raju; Selvaraj, Gopalan

    2003-11-25

    Pollen fecundity is crucial to crop productivity and also to biodiversity in general. Pollen development is supported by the tapetum, a metabolically active sporophytic nurse layer that devotes itself to this process. The tapetum in cereals and a vast majority of other plants is of the nonamoeboid type. Unable to reach out to microspores, it secretes nutrients into the anther locule where the microspores reside and develop. Orbicules (Ubisch bodies), studied in various plants since their discovery approximately 140 years ago, are a hallmark of the secretory tapetum. Their significance to tapetal or pollen development has not been established. We have identified in wheat and rice an anther-specific single-copy gene (per haploid genome equivalent) whose suppression in rice by RNA interference nearly eliminated the seed set. The flowers in the transgenics were normal for female functions, but the pollen collapsed and became less viable. Further characterization of the gene product, named RAFTIN, in wheat has shown that it is present in pro-orbicule bodies and it is accumulated in Ubisch bodies. Furthermore, it is targeted to microspore exine. Although the carboxyl portion of RAFTINs shares short, dispersed amino acid sequences (BURP domain) in common with a variety of proteins of disparate biological contexts, the occurrence RAFTIN per se is limited to cereals; neither the Arabidopsis genome nor the vast collection of ESTs suggests any obvious dicot homologs. Furthermore, our results show that RAFTIN is essential for the late phase of pollen development in cereals. PMID:14612572

  2. Maize opaque10 Encodes a Cereal-Specific Protein That Is Essential for the Proper Distribution of Zeins in Endosperm Protein Bodies.

    Science.gov (United States)

    Yao, Dongsheng; Qi, Weiwei; Li, Xia; Yang, Qing; Yan, Shumei; Ling, Huiling; Wang, Gang; Wang, Guifeng; Song, Rentao

    2016-08-01

    Cereal storage proteins are major nitrogen sources for humans and livestock. Prolamins are the most abundant storage protein in most cereals. They are deposited into protein bodies (PBs) in seed endosperm. The inner structure and the storage mechanism for prolamin PBs is poorly understood. Maize opaque10 (o10) is a classic opaque endosperm mutant with misshapen PBs. Through positional cloning, we found that O10 encodes a novel cereal-specific PB protein. Its middle domain contains a seven-repeat sequence that is responsible for its dimerization. Its C terminus contains a transmembrane motif that is required for its ER localization and PB deposition. A cellular fractionation assay indicated that O10 is initially synthesized in the cytoplasm and then anchored to the ER and eventually deposited in the PB. O10 can interact with 19-kD and 22-kD α-zeins and 16-kD and 50-kD γ-zeins through its N-terminal domain. An immunolocalization assay indicated that O10 co-localizes with 16-kD γ-zein and 22-kD α-zein in PBs, forming a ring-shaped structure at the interface between the α-zein-rich core and the γ-zein-rich peripheral region. The loss of O10 function disrupts this ring-shaped distribution of 22-kD and 16-kD zeins, resulting in misshapen PBs. These results showed that O10, as a newly evolved PB protein, is essential for the ring-shaped distribution of 22-kD and 16-kD zeins and controls PB morphology in maize endosperm. PMID:27541862

  3. Identification of Misfolded Proteins in Body Fluids for the Diagnosis of Prion Diseases

    OpenAIRE

    Francesca Properzi; Maurizio Pocchiari

    2013-01-01

    Transmissible spongiform encephalopathy (TSE) or prion diseases are fatal rare neurodegenerative disorders affecting man and animals and caused by a transmissible infectious agent. TSE diseases are characterized by spongiform brain lesions with neuronal loss and the abnormal deposition in the CNS, and to less extent in other tissues, of an insoluble and protease resistant form of the cellular prion protein (PrPC), named PrPTSE. In man, TSE diseases affect usually people over 60 years of age w...

  4. Plant Cell Wall Proteins: A Large Body of Data, but What about Runaways?

    OpenAIRE

    Cécile Albenne; Hervé Canut; Laurent Hoffmann; Elisabeth Jamet

    2014-01-01

    Plant cell wall proteomics has been a very dynamic field of research for about fifteen years. A full range of strategies has been proposed to increase the number of identified proteins and to characterize their post-translational modifications. The protocols are still improving to enlarge the coverage of cell wall proteomes. Comparisons between these proteomes have been done based on various working strategies or different physiological stages. In this review, two points are highlighted. The ...

  5. Reconstitution of translation from Thermus thermophilus reveals a minimal set of components sufficient for protein synthesis at high temperatures and functional conservation of modern and ancient translation components

    OpenAIRE

    Ying ZHOU; Asahara, Haruichi; Gaucher, Eric A.; Chong, Shaorong

    2012-01-01

    Thermus thermophilus is a thermophilic model organism distantly related to the mesophilic model organism E. coli. We reconstituted protein translation of Thermus thermophilus in vitro from purified ribosomes, transfer ribonucleic acids (tRNAs) and 33 recombinant proteins. This reconstituted system was fully functional, capable of translating natural messenger RNA (mRNA) into active full-length proteins at temperatures up to 65°C and with yields up to 60 μg/ml. Surprisingly, the synthesis of a...

  6. Isolation of soybean protein P34 from oil bodies using hydrophobic interaction chromatography

    Directory of Open Access Journals (Sweden)

    Seidel-Morgenstern Andreas

    2008-03-01

    Full Text Available Abstract Background Soybeans play a prominent role in allergologic research due to the high incidence of allergic reactions. For detailed studies on specific proteins it is necessary to have access to a large amount of pure substance. Results In this contribution, a method for purifying soybean (Glycine max protein P34 (also called Gly m Bd 30 K or Gly m 1 using hydrophobic interaction chromatography is presented. After screening experiments using 1 mL HiTrap columns, Butyl Sepharose 4 FF was selected for further systematic investigations. With this stationary phase, suitable operation conditions for two-step gradient elution using ammonium sulphate were determined experimentally. The separation conditions obtained in a small column could be scaled up successfully to column volumes of 7.5 and 75 mL, allowing for high product purities of almost 100% with a yield of 27% for the chromatographic separation step. Conditions could be simplified further using a onestep gradient, which gave comparable purification in a shorter process time. The identity of the purified protein was verified using in-gel digestion and mass spectrometry as well as immunological techniques. Conclusion With the technique presented it is possible to produce, within a short timeframe, pure P34, suitable for further studies where an example antigen is needed.

  7. Crescimento relativo dos componentes do corpo de zebuínos de diferentes raças Relative growth of body components from different zebu breeds

    Directory of Open Access Journals (Sweden)

    André Mendes Jorge

    2000-05-01

    Full Text Available Foram utilizados 64 tourinhos Gir, Guzerá, Mocho Tabapuã e Nelore, com 24 meses e pesos vivos médios iniciais de 376,4; 357,6, 362,0 e 368,6 kg respectivamente. Os animais de cada raça foram divididos em cinco categorias. Uma categoria foi abatida imediatamente (AB, três receberam, em baias individuais, ração contendo 50% de concentrado na matéria seca ad libitum (I, II e III e uma recebeu a mesma ração, em quantidade restrita (AR. Os animais I, II e III foram abatidos ao atingirem pesos vivos individuais de 405, 450 e 495 kg, respectivamente. No abate determinou-se o peso corporal vazio (PCVZ, e pesaram-se a cabeça, os pés, o couro, os órgãos internos e as vísceras. Adotou-se a equação de regressão do logaritmo do peso de cada componente do corpo em função do logaritmo do PCVZ. Todos os componentes estudados, excetuando-se o fígado e o baço, revelaram um menor ímpeto de crescimento em relação ao do PCVZ.Sixty three steers from zebu breeds Gyr, Guzerat, Mocho Tabapua and Nellore, weighing, respectively, 376.4, 357.6, 362.0 and 368.6 kg initial live weight and twenty-four months old, were used. The animals from each breed were divided into five groups. One group was ramdomly assigned to immediate slaughter (AB, three groups were full-fed a ration containing 50% concentrate, dry matter basis until reaching the slaughter weights of 405, 450 and 495 kg, respectively (group I, II and III. The fifth group was fed a restricted amount of the same ration (AR. At slaughter the empty body weight (EBW was determined and the weight of internal organs and viscera were recorded. Regression equations of log weight of body component as a function of log EBW were fitted. All body components studied, with exception of liver and spleen, developed slowlier than EBW.

  8. Benefits of Whole-Body Vibration, as a Component of the Pulmonary Rehabilitation, in Patients with Chronic Obstructive Pulmonary Disease: A Narrative Review with a Suitable Approach

    Science.gov (United States)

    Sá-Caputo, Danubia; Gonçalves, Cintia Renata; Morel, Danielle Soares; Marconi, Eloá Moreira; Fróes, Patrícia; Rufino, Rogério; Costa, Cláudia Henrique; Lopes, Agnaldo José; Arnóbio, Adriano; Asad, Nasser Ribeiro; Marin, Pedro Jesus; Furness, Trentham; Bernardo-Filho, Mario

    2016-01-01

    Background. Appropriate management, including pulmonary rehabilitation, associated with correct diagnosis of chronic obstructive pulmonary disease (COPD) in patients can contribute to improving clinical conditions of these patients. Physical activity is recommended for COPD patients. Whole-body vibration (WBV) is a modality of physical activity. Putting together the biological effects and safe use of WBV, it may be a potentially feasible intervention to add to pulmonary rehabilitation. The purpose of this investigation was to systematically review studies regarding the effects of WBV, as a component of the pulmonary rehabilitation, in patients with COPD. Results. A total of six publications met inclusion for review. There was evidence to support the beneficial use of WBV to improve functional performance of the lower limbs and quality of life. However, the appropriateness of and descriptors of WBV methods were poorly described. Conclusions. The results of this review support the use of WBV as a component of pulmonary rehabilitation to assist management of patients with COPD. However, future research should examine the dose-response curve and optimal dosing regimen of WBV according to standard reporting recommendations for people with COPD. Such an approach will allow comparison among studies and the potential of meta-analysis of randomized controlled trials. PMID:27190529

  9. G-protein signalling components GCR1 and GPA1 mediate responses to multiple abiotic stresses in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Navjyoti eChakraborty

    2015-11-01

    Full Text Available G-protein signalling components have been implicated in some individual stress responses in Arabidopsis, but have not been comprehensively evaluated at the genetic and biochemical level. Stress emerged as the largest functional category in our whole transcriptome analyses of knock-out mutants of GCR1 and/or GPA1 in Arabidopsis (Chakraborty et al., 2015a, PloS one 10, e0117819 and Chakraborty et al., 2015b, Plant Mol. Biol., doi: 10.1007/s11103-015-0374-2. This led us to ask whether G-protein signalling components offer converging points in the plant’s response to multiple abiotic stresses. In order to test this hypothesis, we carried out detailed analysis of the stress category in the present study, which revealed 144 differentially expressed genes (DEGs, spanning a wide range of abiotic stresses, including heat, cold, salt, light stress etc. Only 10 of these DEGs are shared by all the three mutants, while the single mutants (GCR1/GPA1 shared more DEGs between themselves than with the double mutant (GCR1-GPA1. RT-qPCR validation of 28 of these genes spanning different stresses revealed identical regulation of the DEGs shared between the mutants. We also validated the effects of cold, heat and salt stresses in all the 3 mutants and WT on % germination, root and shoot length, relative water content, proline content, lipid peroxidation and activities of catalase, ascorbate peroxidase and superoxide dismutase. All the 3 mutants showed evidence of stress tolerance, especially to cold, followed by heat and salt, in terms of all the above parameters. This clearly shows the role of GCR1 and GPA1 in mediating the plant’s response to multiple abiotic stresses for the first time, especially cold, heat and salt stresses. This also implies a role for classical G-protein signalling pathways in stress sensitivity in the normal plants of Arabidopsis. This is also the first genetic and biochemical evidence of abiotic stress tolerance rendered by knock

  10. Protein-Pacing and Multi-Component Exercise Training Improves Physical Performance Outcomes in Exercise-Trained Women: The PRISE 3 Study.

    Science.gov (United States)

    Arciero, Paul J; Ives, Stephen J; Norton, Chelsea; Escudero, Daniela; Minicucci, Olivia; O'Brien, Gabe; Paul, Maia; Ormsbee, Michael J; Miller, Vincent; Sheridan, Caitlin; He, Feng

    2016-01-01

    The beneficial cardiometabolic and body composition effects of combined protein-pacing (P; 5-6 meals/day at 2.0 g/kg BW/day) and multi-mode exercise (resistance, interval, stretching, endurance; RISE) training (PRISE) in obese adults has previously been established. The current study examines PRISE on physical performance (endurance, strength and power) outcomes in healthy, physically active women. Thirty exercise-trained women (>4 days exercise/week) were randomized to either PRISE (n = 15) or a control (CON, 5-6 meals/day at 1.0 g/kg BW/day; n = 15) for 12 weeks. Muscular strength (1-RM bench press, 1-RM BP) endurance (sit-ups, SUs; push-ups, PUs), power (bench throws, BTs), blood pressure (BP), augmentation index, (AIx), and abdominal fat mass were assessed at Weeks 0 (pre) and 13 (post). At baseline, no differences existed between groups. Following the 12-week intervention, PRISE had greater gains (p < 0.05) in SUs, PUs (6 ± 7 vs. 10 ± 7, 40%; 8 ± 13 vs. 14 ± 12, 43% ∆reps, respectively), BTs (11 ± 35 vs. 44 ± 34, 75% ∆watts), AIx (1 ± 9 vs. -5 ± 11, 120%), and DBP (-5 ± 9 vs. -11 ± 11, 55% ∆mmHg). These findings suggest that combined protein-pacing (P; 5-6 meals/day at 2.0 g/kg BW/day) diet and multi-component exercise (RISE) training (PRISE) enhances muscular endurance, strength, power, and cardiovascular health in exercise-trained, active women. PMID:27258301

  11. PPI layouts: BioJS components for the display of Protein-Protein Interactions [v1; ref status: indexed, http://f1000r.es/2u5

    Directory of Open Access Journals (Sweden)

    Gustavo A. Salazar

    2014-02-01

    Full Text Available Summary: We present two web-based components for the display of Protein-Protein Interaction networks using different self-organizing layout methods: force-directed and circular. These components conform to the BioJS standard and can be rendered in an HTML5-compliant browser without the need for third-party plugins. We provide examples of interaction networks and how the components can be used to visualize them, and refer to a more complex tool that uses these components. Availability: http://github.com/biojs/biojs; http://dx.doi.org/10.5281/zenodo.7753

  12. Effect of Borax and cysteamine compound on finishing sheep growth performance, nitrogen retention, serum biochemical indices and body protein motabolism

    International Nuclear Information System (INIS)

    Experiment was conducted to investigate the effects of borax, cysteamine and the mixture of two compounds on growth performance, serum biochemical indices, protein turn-over and nitrogen aggradation of sheep by using isotope (15N-Gly) technique, nitrogen balance trial and serum testing. The results showed that both borax and cysteamine could increase the growth performance of sheep, especially the cysteamine and its mixture with borax, which increased average daily feed intake (P3) and levothyroxine (T4) of mixture were higher than that of control group (P<0.05), and the concentrations of growth hormone (GH) and insulin-like growth factors (IGF-1) were also significantly higher than those of control group (P<0.01). There was no significant difference of insulin (INS) between experiment groups and control group (P<0.05). Both mixture and borax contributed to increasing nitrogen retention, net nitrogen utilization, digestibility and biological value. Both borax and cysteamine accelerated protein degradation rate, apparent amino acid utilization rate and net amino acid utilization rate as well as biological value, body protein and oxidation rate, but the former was greater than the later. (authors)

  13. Refolding and purification of recombinant L-asparaginase from inclusion bodies of E. coli into active tetrameric protein.

    Science.gov (United States)

    Upadhyay, Arun K; Singh, Anupam; Mukherjee, K J; Panda, Amulya K

    2014-01-01

    A tetrameric protein of therapeutic importance, Escherichia coli L-asparaginase-II was expressed in Escherichia coli as inclusion bodies (IBs). Asparaginase IBs were solubilized using low concentration of urea and refolded into active tetrameric protein using pulsatile dilution method. Refolded asparaginase was purified in two steps by ion-exchange and gel filtration chromatographic techniques. The recovery of bioactive asparaginase from IBs was around 50%. The melting temperature (Tm) of the purified asparaginase was found to be 64°C. The specific activity of refolded, purified asparaginase was found to be comparable to the commercial asparaginase (190 IU/mg). Enzymatic activity of the refolded asparaginase was high even at four molar urea solutions, where the IB aggregates are completely solubilized. From the comparison of chemical denaturation data and activity at different concentrations of guanidine hydrochloride, it was observed that dissociation of monomeric units precedes the complete loss of helical secondary structures. Protection of the existing native-like protein structure during solubilization of IB aggregates with 4 M urea improved the propensity of monomer units to form oligomeric structure. Our mild solubilization technique retaining native-like structures, improved recovery of asparaginase in bioactive tetrameric form. PMID:25309524

  14. Cerebellar distribution of calcitonin gene-related peptide (CGRP) and its receptor components calcitonin receptor-like receptor (CLR) and receptor activity modifying protein 1 (RAMP1) in rat

    DEFF Research Database (Denmark)

    Edvinsson, Lars; Eftekhari, Sajedeh; Salvatore, Christopher A;

    2011-01-01

    Clinical and experimental results have revealed a fundamental role of calcitonin gene-related peptide (CGRP) in primary headaches. CGRP is widely expressed in neurons both in the central nervous system (CNS) and in peripheral sensory nerves. In the CNS there is a wide distribution of CGRP...... modifying protein type 1 (RAMP1) have been developed. In the present study we therefore examined immunohistochemically the distribution of CGRP and its receptor components in the cerebellum. CGRP immunoreactivity was only found intracellularly in the cerebellar Purkinje cell bodies, whereas CLR and RAMP1...... were detected on the surface of the Purkinje cell bodies and in their processes. The elaborate dendritic tree of Purkinje cell fibers was distinctly visualized with the RAMP1 antibody. In addition, profoundly stained fibers spanning from the molecular layer into the medulla was observed with the RAMP1...

  15. Preventive Effects of Chitosan Coacervate Whey Protein on Body Composition and Immunometabolic Aspect in Obese Mice

    Directory of Open Access Journals (Sweden)

    Gabriel Inácio de Morais Honorato de Souza

    2014-01-01

    Full Text Available Functional foods containing bioactive compounds of whey may play an important role in prevention and treatment of obesity. The aim of this study was to investigate the prospects of the biotechnological process of coacervation of whey proteins (CWP in chitosan and test its antiobesogenic potential. Methods. CWP (100 mg·kg·day was administered in mice with diet-induced obesity for 8 weeks. The animals were divided into four groups: control normocaloric diet gavage with water (C or coacervate (C-CWP, and high fat diet gavage with water (HF or coacervate (HF-CWP. Results. HF-CWP reduced weight gain and serum lipid fractions and displayed reduced adiposity and insulin. Adiponectin was significantly higher in HF-CWP group when compared to the HF. The level of LPS in HF-W group was significantly higher when compared to HF-CWP. The IL-10 showed an inverse correlation between the levels of insulin and glucose in the mesenteric adipose tissue in the HF-CWP group. CWP promoted an increase in both phosphorylation AMPK and the amount of ATGL in the mesenteric adipose tissue in HF-CWP group. Conclusion. CWP was able to modulate effects, possibly due to its high biological value of proteins. We observed a protective effect against obesity and improved the inflammatory milieu of white adipose tissue.

  16. Development of methods for body composition studies

    Energy Technology Data Exchange (ETDEWEB)

    Mattsson, Soeren [Department of Radiation Physics, Lund University, Malmoe University Hospital, SE-205 02 Malmoe (Sweden); Thomas, Brian J [School of Physical and Chemical Sciences, Queensland University of Technology, Brisbane, QLD 4001 (Australia)

    2006-07-07

    This review is focused on experimental methods for determination of the composition of the human body, its organs and tissues. It summarizes the development and current status of fat determinations from body density, total body water determinations through the dilution technique, whole and partial body potassium measurements for body cell mass estimates, in vivo neutron activation analysis for body protein measurements, dual-energy absorptiometry (DEXA), computed tomography (CT) and magnetic resonance imaging (MRI, fMRI) and spectroscopy (MRS) for body composition studies on tissue and organ levels, as well as single- and multiple-frequency bioimpedance (BIA) and anthropometry as simple easily available methods. Methods for trace element analysis in vivo are also described. Using this wide range of measurement methods, together with gradually improved body composition models, it is now possible to quantify a number of body components and follow their changes in health and disease. (review)

  17. Development of methods for body composition studies

    International Nuclear Information System (INIS)

    This review is focused on experimental methods for determination of the composition of the human body, its organs and tissues. It summarizes the development and current status of fat determinations from body density, total body water determinations through the dilution technique, whole and partial body potassium measurements for body cell mass estimates, in vivo neutron activation analysis for body protein measurements, dual-energy absorptiometry (DEXA), computed tomography (CT) and magnetic resonance imaging (MRI, fMRI) and spectroscopy (MRS) for body composition studies on tissue and organ levels, as well as single- and multiple-frequency bioimpedance (BIA) and anthropometry as simple easily available methods. Methods for trace element analysis in vivo are also described. Using this wide range of measurement methods, together with gradually improved body composition models, it is now possible to quantify a number of body components and follow their changes in health and disease. (review)

  18. Effect of induced mutation by space-flight on main agronomic traits and protein components in soybean

    International Nuclear Information System (INIS)

    We selected three soybean varieties with specific quality, Zhonghuang 28, Zhonghuang 29 and Zhonghuang 31 as experimental materials. The seeds were boarded on the recoverable breeding satellite Shijian No. 8for spaceflight treatment and the untreated materials used as controls. Major agronomic traits in SP1 and SP2, quality traits (protein content, fat content, protein component, 11S /7S globulin) in SP2 and SP3 were investigated. The results showed that three soybean varieties gave different response to the space environment. A total of 15 variants were obtained in the SP1 generation with the total variation rate of 0. 649%. 3 variants came from SP1 of Zhonghuang 28, with variation rate of 0. 417%, and the others came from SP1 of Zhonghuang 29, and the variation rate was 1. 667%. No variation was found in SP1 of Zhonghuang 31. Traits segregation was found to occur in 7 variants in SP2 generation. Agronomic traits of variants were found significant difference between SP2 and SP1 generation. The mutation was found in 11S globulin subunit and Kunitz soybean trypsin inhibitor. Space mutation could be one of methods to improve both agronomic and quality traits in soybean breeding. (authors)

  19. Biochemical analysis of the interaction of calcium with toposome: a major protein component of the sea urchin egg and embryo.

    Science.gov (United States)

    Hayley, Michael; Sun, Ming; Merschrod, Erika F; Davis, Philip J; Robinson, John J

    2008-04-01

    We have investigated the biochemical and functional properties of toposome, a major protein component of sea urchin eggs and embryos. Atomic force microscopy was utilized to demonstrate that a Ca(2+)-driven change in secondary structure facilitated toposome binding to a lipid bilayer. Thermal denaturation studies showed that toposome was dependent upon calcium in a manner paralleling the effect of this cation on secondary and tertiary structure. The calcium-induced, secondary, and tertiary structural changes had no effect on the chymotryptic cleavage pattern. However, the digestion pattern of toposome bound to phosphatidyl serine liposomes did vary as a function of calcium concentration. We also investigated the interaction of this protein with various metal ions. Calcium, Mg(2+), Ba(2+), Cd(2+), Mn(2+), and Fe(3+) all bound to toposome. In addition, Cd(2+) and Mn(2+) displaced Ca(2+), prebound to toposome, while Mg(2+), Ba(2+), and Fe(3+) had no effect. Collectively, these results further enhance our understanding of the role of Ca(2+) in modulating the biological activity of toposome. PMID:17786928

  20. Protein-electrophoretic variations detected in the fat body of the solitary form of the desert locust Schistocerca gregaria (Forskal)(Orthoptera: Acrididae)

    International Nuclear Information System (INIS)

    The polyacrylamide gel electrophoresis (PAGE) was performed on the fat body tissues of the solitary form of the 5th nymph instar as well as three different phases of adult males and females namely newly emerged, yellow and non-yellow males (mature and nonmature), before oviposition and after oviposition females of Schistocerca gregaria. The total number of protein bands detected in the fat body of the 5th nymph instar amounted to 25 bands. Meanwhile, those detected in the fat body of newly emerged males, non-mature males and mature males were 22, 19, and 21 respectively. On the other hand, the numbers of protein bands detected in the newly emerged females were 28 bands. These bands varied in the successive female phases to be 21 and 30 bands before oviposition and after oviposition respectively. One protein band was detected in males only and considered as a male specific protein ( M-sp.) whereas, four bands were detected in females only and considered as female specific proteins ( F-sp.). Ten protein bands were considered as major fat body protein bands since they were commonly detected in all the investigated phases.

  1. MAGGIE Component 1: Identification and Purification of Native and Recombinant Multiprotein Complexes and Modified Proteins from Pyrococcus furiosus

    Energy Technology Data Exchange (ETDEWEB)

    Adams, Michael W. [University of Georgia; W. W. Adams, Michael

    2014-01-07

    Virtualy all cellular processes are carried out by dynamic molecular assemblies or multiprotein complexes (PCs), the composition of which is largely unknown. Structural genomics efforts have demonstrated that less than 25% of the genes in a given prokaryotic genome will yield stable, soluble proteins when expressed using a one-ORF-at-a-time approach. We proposed that much of the remaining 75% of the genes encode proteins that are part of multiprotein complexes or are modified post-translationally, for example, with metals. The problem is that PCs and metalloproteins (MPs) cannot be accurately predicted on a genome-wide scale. The only solution to this dilemma is to experimentally determine PCs and MPs in biomass of a model organism and to develop analytical tools that can then be applied to the biomass of any other organism. In other words, organisms themselves must be analyzed to identify their PCs and MPs: “native proteomes” must be determined. This information can then be utilized to design multiple ORF expression systems to produce recombinant forms of PCs and MPs. Moreover, the information and utility of this approach can be enhanced by using a hyperthermophile, one that grows optimally at 100°C, as a model organism. By analyzing the native proteome at close to 100 °C below the optimum growth temperature, we will trap reversible and dynamic complexes, thereby enabling their identification, purification, and subsequent characterization. The model organism for the current study is Pyrococcus furiosus, a hyperthermophilic archaeon that grows optimally at 100°C. It is grown up to 600-liter scale and kg quantities of biomass are available. In this project we identified native PCs and MPs using P. furiosus biomass (with MS/MS analyses to identify proteins by component 4). In addition, we provided samples of abundant native PCs and MPs for structural characterization (using SAXS by component 5). We also designed and evaluated generic bioinformatics and

  2. Generation of transgenic cynomolgus monkeys that express green fluorescent protein throughout the whole body.

    Science.gov (United States)

    Seita, Yasunari; Tsukiyama, Tomoyuki; Iwatani, Chizuru; Tsuchiya, Hideaki; Matsushita, Jun; Azami, Takuya; Okahara, Junko; Nakamura, Shinichiro; Hayashi, Yoshitaka; Hitoshi, Seiji; Itoh, Yasushi; Imamura, Takeshi; Nishimura, Masaki; Tooyama, Ikuo; Miyoshi, Hiroyuki; Saitou, Mitinori; Ogasawara, Kazumasa; Sasaki, Erika; Ema, Masatsugu

    2016-01-01

    Nonhuman primates are valuable for human disease modelling, because rodents poorly recapitulate some human diseases such as Parkinson's disease and Alzheimer's disease amongst others. Here, we report for the first time, the generation of green fluorescent protein (GFP) transgenic cynomolgus monkeys by lentivirus infection. Our data show that the use of a human cytomegalovirus immediate-early enhancer and chicken beta actin promoter (CAG) directed the ubiquitous expression of the transgene in cynomolgus monkeys. We also found that injection into mature oocytes before fertilization achieved homogenous expression of GFP in each tissue, including the amnion, and fibroblasts, whereas injection into fertilized oocytes generated a transgenic cynomolgus monkey with mosaic GFP expression. Thus, the injection timing was important to create transgenic cynomolgus monkeys that expressed GFP homogenously in each of the various tissues. The strategy established in this work will be useful for the generation of transgenic cynomolgus monkeys for transplantation studies as well as biomedical research. PMID:27109065

  3. Nuclear LSm8 affects number of cytoplasmic processing bodies via controlling cellular distribution of Like-Sm proteins

    Czech Academy of Sciences Publication Activity Database

    Novotný, Ivan; Podolská, Kateřina; Blažíková, Michaela; Valášek, Leoš; Svoboda, Petr; Staněk, David

    2012-01-01

    Roč. 23, č. 19 (2012), s. 3776-3785. ISSN 1059-1524 R&D Projects: GA AV ČR KAN200520801; GA ČR GA204/07/0133; GA ČR GAP305/10/2215; GA ČR GAP302/11/1910; GA ČR(CZ) GBP305/12/G034 Institutional research plan: CEZ:AV0Z50390703; CEZ:AV0Z50520514; CEZ:AV0Z50200510 Institutional support: RVO:68378050 ; RVO:68378041 ; RVO:61388971 Keywords : P-bodies * LSm proteins * mRNA degradation Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.604, year: 2012

  4. Body Mass Parameters, Lipid Profiles and Protein Contents of Zebrafish Embryos and Effects of 2,4-Dinitrophenol Exposure

    Science.gov (United States)

    Hachicho, Nancy; Reithel, Sarah; Miltner, Anja; Heipieper, Hermann J.; Küster, Eberhard; Luckenbach, Till

    2015-01-01

    Morphology and physiology of fish embryos undergo dramatic changes during their development until the onset of feeding, supplied only by endogenous yolk reserves. For obtaining an insight how these restructuring processes are reflected by body mass related parameters, dry weights (dw), contents of the elements carbon and nitrogen and lipid and protein levels were quantified in different stages within the first four days of embryo development of the zebrafish (Danio rerio). The data show age dependent changes in tissue composition. Dry weights decreased significantly from 79μgdw/egg at 0hours post fertilization (hpf) to 61 μgdw/egg after 96 hpf. The amounts of total carbon fluctuated between 460 mg g-1 and 540 mg g-1 dw, nitrogen was at about 100 mg g-1 dw and total fatty acids were between 48–73 mg g-1 dw. In contrast to these parameters that remained relatively constant, the protein content, which was 240 mg g-1 at 0 hpf, showed an overall increase of about 40%. Comparisons of intact eggs and dechorionated embryos at stages prior to hatching (24, 30, 48 hpf) showed that the differences seen for dry weight and for carbon and nitrogen contents became smaller at more advanced stages, consistent with transition of material from the chorion to embryo tissue. Further, we determined the effect of 2,4-dinitrophenol at a subacutely toxic concentration (14 μM, LC10) as a model chemical challenge on the examined body mass related parameters. The compound caused significant decreases in phospholipid and glycolipid fatty acid contents along with a decrease in the phospholipid fatty acid unsaturation index. No major changes were observed for the other examined parameters. Lipidomic studies as performed here may thus be useful for determining subacute effects of lipophilic organic compounds on lipid metabolism and on cellular membranes of zebrafish embryos. PMID:26292096

  5. Identification of G Protein-Coupled Receptors (GPCRs in Primary Cilia and Their Possible Involvement in Body Weight Control.

    Directory of Open Access Journals (Sweden)

    Yoshihiro Omori

    Full Text Available Primary cilia are sensory organelles that harbor various receptors such as G protein-coupled receptors (GPCRs. We analyzed subcellular localization of 138 non-odorant GPCRs. We transfected GPCR expression vectors into NIH3T3 cells, induced ciliogenesis by serum starvation, and observed subcellular localization of GPCRs by immunofluorescent staining. We found that several GPCRs whose ligands are involved in feeding behavior, including prolactin-releasing hormone receptor (PRLHR, neuropeptide FF receptor 1 (NPFFR1, and neuromedin U receptor 1 (NMUR1, localized to the primary cilia. In addition, we found that a short form of dopamine receptor D2 (DRD2S is efficiently transported to the primary cilia, while a long form of dopamine receptor D2 (DRD2L is rarely transported to the primary cilia. Using an anti-Prlhr antibody, we found that Prlhr localized to the cilia on the surface of the third ventricle in the vicinity of the hypothalamic periventricular nucleus. We generated the Npy2r-Cre transgenic mouse line in which Cre-recombinase is expressed under the control of the promoter of Npy2r encoding a ciliary GPCR. By mating Npy2r-Cre mice with Ift80 flox mice, we generated Ift80 conditional knockout (CKO mice in which Npy2r-positive cilia were diminished in number. We found that Ift80 CKO mice exhibited a body weight increase. Our results suggest that Npy2r-positive cilia are important for body weight control.

  6. Identification of G Protein-Coupled Receptors (GPCRs) in Primary Cilia and Their Possible Involvement in Body Weight Control.

    Science.gov (United States)

    Omori, Yoshihiro; Chaya, Taro; Yoshida, Satoyo; Irie, Shoichi; Tsujii, Toshinori; Furukawa, Takahisa

    2015-01-01

    Primary cilia are sensory organelles that harbor various receptors such as G protein-coupled receptors (GPCRs). We analyzed subcellular localization of 138 non-odorant GPCRs. We transfected GPCR expression vectors into NIH3T3 cells, induced ciliogenesis by serum starvation, and observed subcellular localization of GPCRs by immunofluorescent staining. We found that several GPCRs whose ligands are involved in feeding behavior, including prolactin-releasing hormone receptor (PRLHR), neuropeptide FF receptor 1 (NPFFR1), and neuromedin U receptor 1 (NMUR1), localized to the primary cilia. In addition, we found that a short form of dopamine receptor D2 (DRD2S) is efficiently transported to the primary cilia, while a long form of dopamine receptor D2 (DRD2L) is rarely transported to the primary cilia. Using an anti-Prlhr antibody, we found that Prlhr localized to the cilia on the surface of the third ventricle in the vicinity of the hypothalamic periventricular nucleus. We generated the Npy2r-Cre transgenic mouse line in which Cre-recombinase is expressed under the control of the promoter of Npy2r encoding a ciliary GPCR. By mating Npy2r-Cre mice with Ift80 flox mice, we generated Ift80 conditional knockout (CKO) mice in which Npy2r-positive cilia were diminished in number. We found that Ift80 CKO mice exhibited a body weight increase. Our results suggest that Npy2r-positive cilia are important for body weight control. PMID:26053317

  7. Predicting the image noise level of prospective ECG-triggered coronary computed tomography angiography: quantitative measurement of thoracic component versus body mass index.

    Science.gov (United States)

    Kim, Hyeongmin; Park, Chul Hwan; Han, Kyung Hwa; Kim, Tae Hoon

    2015-12-01

    We evaluated the feasibility of using quantitatively measured thoracic components, as compared to body mass index (BMI), for predicting the image noise of coronary computed tomography angiography (CCTA). One hundred subjects (M:F = 64:36; mean age, 55 ± 8.8 years) who underwent prospective electrocardiography-gated CCTA and low-dose chest computed tomography (CT) were analyzed retrospectively. The image noise of the CCTA was determined by the standard deviation of the attenuation value in a region of interest on the aortic root level. On the low-dose chest CT, the areas of the thoracic components were measured at the aortic root level. An auto-segmentation technique with the following threshold levels was used: quantitatively measured area of total thorax [QMAtotal: -910 to 1000 Hounsfield units (HU)], lung (QMAlung: -910 to -200 HU), fat (QMAfat: -200 to 0 HU), muscle (QMAmuscle: 0-300 HU), soft tissue (fat + muscle, QMAsoft tissue: -200 to 300 HU), bone (QMAbone: 300-1000 HU) and solid tissue (fat + muscle + bone, QMAsolid tissue: -200 to 1000 HU). The relationship between image noise and variable biometric parameters including QMA was analyzed, and the linear correlation coefficients were used as indicators of the strength of association. Among the variable biometric parameters, including BMI, QMAsolid tissue showed the highest correlation coefficient with image noise in all subjects (r = 0.804), males (r = 0.716), females (r = 0.889), the overweight (r = 0.556), and the non-overweight subgroups (r = 0.783). QMAsolid tissue can be used as a potential surrogate predictor of the image noise level in low tube voltage CCTA. PMID:26507324

  8. Determination of solvent thinner components in human body fluids by capillary gas chromatography with trapping at low oven temperature for headspace samples.

    Science.gov (United States)

    Lee, X P; Kumazawa, T; Sato, K; Watanabe, K; Seno, H; Suzuki, O

    1998-01-01

    A simple and sensitive method is presented for determination of solvent thinner components in human body fluids by capillary gas chromatography (GC) with a low oven temperature for trapping headspace vapor components. After heating a blood or urine sample containing ethyl acetate, benzene, butan-1-ol, toluene, butyl acetate, isoamyl acetate and ethylbenzene (internal standard) in a 7.5 ml vial at 90 degrees C for 30 min, 5 ml of headspace vapor were drawn into a glass syringe. All vapor was introduced through an injection port in the splitless mode into a DB-624 medium-bore capillary column at a 5 degrees C oven temperature for trapping the volatile compounds, and the oven temperature was programmed up to 110 degrees C for their detection by GC. These conditions gave sharp peaks, a good separation of each peak and low background noise for both whole blood and urine samples. As much as 3.58-55.1 and 3.52-57.9% of the six compounds, which had been added to vials, could be introduced to the GC instrument for whole blood and urine, respectively. The intra-day RSD values in terms of the introduction rate (net recovery) of the six compounds in whole blood and urine samples were < or = 8.1%. The calibration curves showed linearity in the range 0.78-400 ng per 0.5 ml whole blood or urine. The detection limits were 0.5-5 ng per 0.5 ml. The data on toluene in post mortem blood in an actual case are also presented. PMID:9581030

  9. The effect of dietary protein on reproduction in the mare. II. Growth of foals, body mass of mares and serum protein concentration of mares during the anovulatory, transitional and pregnant periods

    OpenAIRE

    F.E. Van Niekerk; C.H. Van Niekerk

    1997-01-01

    The effect of 4 different diets, in terms of protein quantity and quality, on total serum protein (TSP), albumin and globulin was investigated. Non-pregnant mares that were not lactating (n = 36), pregnant mares that had foaled (n = 24) and their foals (n = 24) were used in this study. Daily total protein intake had no effect on blood protein concentrations in the mares. Total protein intake and quality (available essential amino-acids) did affect the body mass of mares during lactation. When...

  10. Regulation of spindle pole body assembly and cytokinesis by the centrin-binding protein Sfi1 in fission yeast.

    Science.gov (United States)

    Lee, I-Ju; Wang, Ning; Hu, Wen; Schott, Kersey; Bähler, Jürg; Giddings, Thomas H; Pringle, John R; Du, Li-Lin; Wu, Jian-Qiu

    2014-09-15

    Centrosomes play critical roles in the cell division cycle and ciliogenesis. Sfi1 is a centrin-binding protein conserved from yeast to humans. Budding yeast Sfi1 is essential for the initiation of spindle pole body (SPB; yeast centrosome) duplication. However, the recruitment and partitioning of Sfi1 to centrosomal structures have never been fully investigated in any organism, and the presumed importance of the conserved tryptophans in the internal repeats of Sfi1 remains untested. Here we report that in fission yeast, instead of doubling abruptly at the initiation of SPB duplication and remaining at a constant level thereafter, Sfi1 is gradually recruited to SPBs throughout the cell cycle. Like an sfi1Δ mutant, a Trp-to-Arg mutant (sfi1-M46) forms monopolar spindles and exhibits mitosis and cytokinesis defects. Sfi1-M46 protein associates preferentially with one of the two daughter SPBs during mitosis, resulting in a failure of new SPB assembly in the SPB receiving insufficient Sfi1. Although all five conserved tryptophans tested are involved in Sfi1 partitioning, the importance of the individual repeats in Sfi1 differs. In summary, our results reveal a link between the conserved tryptophans and Sfi1 partitioning and suggest a revision of the model for SPB assembly. PMID:25031431

  11. Three-dimensional chitin rings from body segments of a pet diplopod species: Characterization and protein interaction studies.

    Science.gov (United States)

    Kaya, Murat; Mulerčikas, Povilas; Sargin, Idris; Kazlauskaitė, Sonata; Baublys, Vykintas; Akyuz, Bahar; Bulut, Esra; Tubelytė, Vaida

    2016-11-01

    Physicochemical characterization of new chitin isolates can provide valuable insights into designing of biomimetic materials. Chitin isolates with a definite three-dimensional (3D) structure can exhibit characteristics that distinguish them from other chitin specimens that are in form of powder or flakes without a definite and uniform shape. Herein, 3D chitin rings were produced from body segments of a diplopod (Archispirostreptus gigas) inhabiting tropical regions. This organism is cultured easily and can reach 38cm in length, which makes it a suitable source for isolation of chitin. The chitin rings were characterized via TGA, FT-IR, SEM and XRD analyses. Enzymatic digestion test with chitinase demonstrated that chitin isolates had high purity (digestion rate: 97.4%). The source organism had high chitin content; 21.02±2.23% on dry weight. Interactions of the chitin rings with bovine serum albumin (BSA) protein were studied under different conditions (pH: 4.0-8.0, chitin amount: 6-14mg, contact time: 30-360min, protein concentration: 0.2-1mg/mL). The highest BSA adsorption was observed at pH5.0 at 20°C. The adsorption equilibrium data exhibited a better fit to Langmuir adsorption and the pseudo-first order kinetic models. The findings presented here can be useful for further studies aiming to develop biocompatible and nontoxic biomaterials. PMID:27524072

  12. Body of Knowledge (BOK) for Leadless Quad Flat No-Lead/Bottom Termination Components (QFN/BTC) Package Trends and Reliability

    Science.gov (United States)

    Ghaffarian, Reza

    2014-01-01

    Bottom terminated components and quad flat no-lead (BTC/QFN) packages have been extensively used by commercial industry for more than a decade. Cost and performance advantages and the closeness of the packages to the boards make them especially unique for radio frequency (RF) applications. A number of high-reliability parts are now available in this style of package configuration. This report presents a summary of literature surveyed and provides a body of knowledge (BOK) gathered on the status of BTC/QFN and their advanced versions of multi-row QFN (MRQFN) packaging technologies. The report provides a comprehensive review of packaging trends and specifications on design, assembly, and reliability. Emphasis is placed on assembly reliability and associated key design and process parameters because they show lower life than standard leaded package assembly under thermal cycling exposures. Inspection of hidden solder joints for assuring quality is challenging and is similar to ball grid arrays (BGAs). Understanding the key BTC/QFN technology trends, applications, processing parameters, workmanship defects, and reliability behavior is important when judicially selecting and narrowing the follow-on packages for evaluation and testing, as well as for the low risk insertion in high-reliability applications.

  13. Dissection of Molecular Mechanisms Regulating Protein Body Formation in Maize Endosperm - DE-FG03-95-ER20183 B139

    Energy Technology Data Exchange (ETDEWEB)

    Brian A. Larkins

    2003-03-21

    Dissection of Molecular Mechanisms Regulating Protein Body Formation in Maize Endosperm - DE-FG03-95-ER20183 Final Technical Report and Patent Summary Dr. Brian A. Larkins, Department of Plant Sciences, University of Arizona, Tucson, AZ 85721 Endosperm texture is an important quality trait in maize, as it influences the shipping characteristics of the grain, its susceptibility to insects, the yield of grits from dry milling, energy costs during wet milling, and the baking and digestibility properties of the flour. There appears to be a causal relationship between kernel hardness and the formation of zein-containing protein bodies, as mutations affecting protein body number and structure are associated with a soft, starchy kernel. In this project we used a variety of approaches to better understand this relationship and investigate the molecular and biochemical changes associated with starchy endosperm mutants. We characterized the distribution of zein mRNAs on endosperm rough endoplasmic reticulum (RER) membranes and the interactions between zein proteins, as each of these could influence the structure of protein bodies. Based on in situ hybridization, mRNAs encoding the 22-kD alpha- and 27-kD gamma-zeins are randomly distributed on RER; hence, mRNA targeting does not appear to influence the formation of protein bodies. Investigation of the interactions between zein proteins (alpha, beta, gamma, delta) with the yeast two-hybrid system showed that interactions between the 19- and 22-alpha-zeins are relatively weak, although each of them interacted strongly with the 10-kD delta-zein. Strong interactions were detected between the alpha- and delta-zeins and the 16-kD gamma- and 15-kD beta-zeins; however, the 50-kD and 27-kD gamma-zeins did not interact detectably with the alpha- and delta-zein proteins. The NH2- and COOH-terminal domains of the 22-kD alpha-zein were found to interact most strongly with the 15-kD beta- and 16-kD gamma-zeins, suggesting the 16-kD and 15

  14. Immunohistochemical Detection of a Unique Protein within Cells of Snakes Having Inclusion Body Disease, a World-Wide Disease Seen in Members of the Families Boidae and Pythonidae

    OpenAIRE

    Chang, Li-Wen; Fu, Ann; Wozniak, Edward; Chow, Marjorie; Duke, Diane G.; Green, Linda; Kelley, Karen; Hernandez, Jorge A; Jacobson, Elliott R.

    2013-01-01

    Inclusion body disease (IBD) is a worldwide disease in captive boa constrictors (boa constrictor) and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s) and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the I...

  15. Model Hirano bodies protect against tau-independent and tau-dependent cell death initiated by the amyloid precursor protein intracellular domain.

    Directory of Open Access Journals (Sweden)

    Matthew Furgerson

    Full Text Available The main pathological hallmarks of Alzheimer's disease are amyloid-beta plaques and neurofibrillary tangles, which are primarily composed of amyloid precursor protein (APP and tau, respectively. These proteins and their role in the mechanism of neurodegeneration have been extensively studied. Hirano bodies are a frequently occurring pathology in Alzheimer's disease as well as other neurodegenerative diseases. However, the physiological role of Hirano bodies in neurodegenerative diseases has yet to be determined. We have established cell culture models to study the role of Hirano bodies in amyloid precursor protein and tau-induced cell death mechanisms. Exogenous expression of APP and either of its c-terminal fragments c31 or Amyloid Precursor Protein Intracellular Domain c58 (AICDc58 enhance cell death. The presence of tau is not required for this enhanced cell death. However, the addition of a hyperphosphorylated tau mimic 352PHPtau significantly increases cell death in the presence of both APP and c31 or AICDc58 alone. The mechanism of cell death induced by APP and its c-terminal fragments and tau was investigated. Fe65, Tip60, p53, and caspases play a role in tau-independent and tau-dependent cell death. In addition, apoptosis was determined to contribute to cell death. The presence of model Hirano bodies protected against cell death, indicating Hirano bodies may play a protective role in neurodegeneration.

  16. Effect of long-term feeding with a plant protein mixture based diet on growth and body/fillet quality traits of large rainbow trout (Oncorhynchus mykiss)

    OpenAIRE

    De Francesco, Matilde; Parisi, Giuliana; Medale, Francoise; Lupi, Paola; Kaushik, Sadasivam; Poli, Bianca

    2004-01-01

    Two diets based oil fish meal (FM) or oil a mixture of plant protein sources supplemented with free amino acids (PP) were fed to rainbow trout having a mean initial body weight of 162.5 g reared at 17 +/- 1 degreesC for 24 weeks to evaluate the effect of plant protein sources oil fish growth, morphological and body quality traits. At the end of the growth study, samples were withdrawn for chemical analyses as well as measurements of different parameters of quality. Compared to the FM gro...

  17. Inclusion body anatomy and functioning of chaperone-mediated in vivo inclusion body disassembly during high-level recombinant protein production in Escherichia coli.

    OpenAIRE

    Rinas, Ursula; Hoffmann, Frank; Betiku, Eriola; Estapé, David; Marten, Sabine

    2007-01-01

    During production in recombinant Escherichia coli, the human basic fibroblast growth factor (hFGF-2) partly aggregates into stable cytoplasmic inclusion bodies. These inclusion bodies additionally contain significant amounts of the heat-shock chaperone DnaK, and putative DnaK substrates such as the elongation factor Tu (ET-Tu) and the metabolic enzymes dihydrolipoamide dehydrogenase (LpdA), tryptophanase (TnaA), and d-tagatose-1,6-bisphosphate aldolase (GatY). Guanidinium hydrochloride induce...

  18. Stable Translocation Intermediates Jam Global Protein Export in Plasmodium falciparum Parasites and Link the PTEX Component EXP2 with Translocation Activity

    Science.gov (United States)

    Mesén-Ramírez, Paolo; Reinsch, Ferdinand; Blancke Soares, Alexandra; Bergmann, Bärbel; Ullrich, Ann-Katrin; Tenzer, Stefan

    2016-01-01

    Protein export is central for the survival and virulence of intracellular P. falciparum blood stage parasites. To reach the host cell, exported proteins cross the parasite plasma membrane (PPM) and the parasite-enclosing parasitophorous vacuole membrane (PVM), a process that requires unfolding, suggestive of protein translocation. Components of a proposed translocon at the PVM termed PTEX are essential in this phase of export but translocation activity has not been shown for the complex and questions have been raised about its proposed membrane pore component EXP2 for which no functional data is available in P. falciparum. It is also unclear how PTEX mediates trafficking of both, soluble as well as transmembrane proteins. Taking advantage of conditionally foldable domains, we here dissected the translocation events in the parasite periphery, showing that two successive translocation steps are needed for the export of transmembrane proteins, one at the PPM and one at the PVM. Our data provide evidence that, depending on the length of the C-terminus of the exported substrate, these steps occur by transient interaction of the PPM and PVM translocon, similar to the situation for protein transport across the mitochondrial membranes. Remarkably, we obtained constructs of exported proteins that remained arrested in the process of being translocated across the PVM. This clogged the translocation pore, prevented the export of all types of exported proteins and, as a result, inhibited parasite growth. The substrates stuck in translocation were found in a complex with the proposed PTEX membrane pore component EXP2, suggesting a role of this protein in translocation. These data for the first time provide evidence for EXP2 to be part of a translocating entity, suggesting that PTEX has translocation activity and provide a mechanistic framework for the transport of soluble as well as transmembrane proteins from the parasite boundary into the host cell. PMID:27168322

  19. Effects of protein deficiency on the rate of radioactivity loss from body constituents in adult rats given 14C-amino acids

    International Nuclear Information System (INIS)

    The effect of protein deficiency on the rate of loss of radioactivity from body constituents was studied in adult rats administered 14C-Chlorella protein hydrolysate or 14C-lysine. Rats were kept on a protein-free diet for 3 weeks and then injected with labelled amino acids and fed on a protein-free diet for 3 more days to allow 14C deposition in tissues. Then they were given experimental diets (protein-free diet, 1% and 10% wheat gluten diets pair-fed with the protein-free diet, and 10% wheat gluten diet ad libitum) for 7 days and sacrificed. The rates of loss of radioactivity from tissue proteins became low in general with the extent of protein deficiency. This increased capacity of tissues to retain 14C-amino acids may result from higher efficiency of protein utilization in protein deficiency. The reutilization of free amino acids and the rate of catabolism of tissue protein are discussed on the basis of the results. The half-life of muscle protein was too long to observe the effects of experimental diets given for 7 days on the rate of loss of radioactivity. (auth.)

  20. Bioconversion of wheat straw and wheat straw components into single-cell protein. [Spicaria fusispora, Cochliobolus specifer, Myrothecium verrucaria, Rhizoctonia solani, and Gliocladium sp

    Energy Technology Data Exchange (ETDEWEB)

    Chahal, D.S.; Moo-Young, M.; Dhillon, G.S.

    1979-01-01

    Several fungi (Aspergillus niger, A. terreus, Cochliobolus specifer, Myrothecium verrucaria, Rhizoctonia solani, Spicaria fusispora, Penicillium sp., and Gliocladium sp.) were isolated from decomposing wheat straw and tested for their ability to utilize whole straw and its components, holocellulose (hemicellulose and cellulose) and cellulose, for the production of single-cell protein (SCP). C. specifer was the most efficient fungus for protein synthesis with the 3 substrates. Using KNO/sub 3/ as N source in mixtures of .04 g N/g substrate (0.04% wt./vol.) at pH 4.5, it was found that incubation periods of 3, 4, and 5 days were optimal for protein production on cellulose and holocellulose fractions, and whole straw, respectively. Whole native straw was the most recalcitrant to bioconversion into SCP; however, protein production was almost doubled when the lignin component was removed using a mixture of NaClO/sub 2/ and HOAc.

  1. A Model of Yeast Cell-Cycle Regulation Based on a Standard Component Modeling Strategy for Protein Regulatory Networks

    Science.gov (United States)

    Laomettachit, Teeraphan; Chen, Katherine C.; Baumann, William T.

    2016-01-01

    To understand the molecular mechanisms that regulate cell cycle progression in eukaryotes, a variety of mathematical modeling approaches have been employed, ranging from Boolean networks and differential equations to stochastic simulations. Each approach has its own characteristic strengths and weaknesses. In this paper, we propose a “standard component” modeling strategy that combines advantageous features of Boolean networks, differential equations and stochastic simulations in a framework that acknowledges the typical sorts of reactions found in protein regulatory networks. Applying this strategy to a comprehensive mechanism of the budding yeast cell cycle, we illustrate the potential value of standard component modeling. The deterministic version of our model reproduces the phenotypic properties of wild-type cells and of 125 mutant strains. The stochastic version of our model reproduces the cell-to-cell variability of wild-type cells and the partial viability of the CLB2-dbΔ clb5Δ mutant strain. Our simulations show that mathematical modeling with “standard components” can capture in quantitative detail many essential properties of cell cycle control in budding yeast. PMID:27187804

  2. Investigation of Interaction of Vaccinia Virus Complement Control Protein and Curcumin with Complement Components C3 and C3b Using Quartz Crystal Microbalance with Dissipation Monitoring Technology

    OpenAIRE

    Kulkarni, Amod P.; Randall, Philippa J.; Murthy, Krishna; Kellaway, Lauriston A; Kotwal, Girish J.

    2010-01-01

    C3 and C3b, the components central to the complement activation, also play a damaging role in several inflammatory disorders. Vaccinia virus complement control protein (VCP) and curcumin (Cur) are natural compounds with different biological origins reported to regulate complement activation. However, both VCP and Cur have not been investigated for their interaction with the third component (C3) prior to it being converted to its activated form (C3b). These two compounds have also not been com...

  3. A direct protein kinase B-targeted anti inflammatory activity of cordycepin from artificially cultured fruit body of Cordyceps militaris

    Directory of Open Access Journals (Sweden)

    Ju Young Yoon

    2015-01-01

    Full Text Available Background: Cordyceps militaris is one of well-known medicinal mushrooms with anti-inflammatory, anti-cancer, anti-diabetic, and anti-obesity activities. Objective: The objective of the following study is to isolate chemical components from the ethanol extract (Cm-EE from Cordyceps militaris and to evaluate their anti-inflammatory activities. Materials and Methods: Column chromatographic separation was performed and anti-inflammatory roles of these compounds were also examined by using NO production and protein kinase B (AKT activity assays. Results: From Cm-EE, 13 constituents, including trehalose (1, cordycepin (2, 6-hydroxyethyladenosine (3, nicotinic amide (4, butyric acid (5, β-dimorphecolic acid (6, α-dimorphecolic acid (7, palmitic acid (8, linoleic acid (9, cordycepeptide A (10, 4-(2-hydroxy-3-((9E,12E-octadeca-9,12-dienoyloxypropoxy-2-(trimethylammoniobutanoate (11, 4-(2-hydroxy-3-(palmitoyloxypropoxy-2-(trimethylammoniobutanoate (12, and linoleic acid methyl ester (13 were isolated. Of these components, compound 2 displayed a significant inhibitory effect on NO production in lipopolysaccharide (LPS-activated RAW264.7 cells. Furthermore, this compound strongly and directly suppressed the kinase activity of AKT, an essential signalling enzyme in LPS-induced NO production, by interacting with its ATP binding site. Conclusion: C. militaris could have anti-inflammatory activity mediated by cordycepin-induced suppression of AKT.

  4. 15N tracer kinetic studies on the validity of various 15N tracer substances for determining whole-body protein parameters in very small preterm infants

    International Nuclear Information System (INIS)

    Reliable 15N tracer substances for tracer kinetic determination of whole-body protein parameters in very small preterm infants are still a matter of intensive research, especially after some doubts have been raised about the validity of [15N]glycine, a commonly used 15N tracer. Protein turnover, synthesis, breakdown, and further protein metabolism data were determined by a paired comparison in four preterm infants. Their post-conceptual age was 32.2 +/- 0.8 weeks, and their body weight was 1670 +/- 181 g. Tracer substances applied in this study were a [15N]amino acid mixture (Ia) and [15N]glycine (Ib). In a second group of three infants with a post conceptual age of 15N-labeled 32.0 +/- 1.0 weeks and a body weight of 1,907 +/- 137 g, yeast protein hydrolysate (II) was used as a tracer substance. A three-pool model was employed for the analysis of the data. This model takes into account renal and fecal 15N losses after a single 15N pulse. Protein turnovers were as follows: 11.9 +/- 3.1 g kg-1 d-1 (Ia), 16.2 +/- 2.5 g kg-1 d-1 (Ib), and 10.8 +/- 3.0 g kg-1 d-1 (II). We were able to demonstrate an overestimation of the protein turnover when Ib was used. There was an expected correspondence in the results obtained from Ia and II. The 15N-labeled yeast protein hydrolysate is a relatively cheap tracer that allows reliable determination of whole-body protein parameters in very small preterm infants

  5. High Whey Protein Intake Delayed the Loss of Lean Body Mass in Healthy Old Rats, whereas Protein Type and Polyphenol/Antioxidant Supplementation Had No Effects

    OpenAIRE

    Laurent Mosoni; Eva Gatineau; Philippe Gatellier; Carole Migné; Isabelle Savary-Auzeloux; Didier Rémond; Emilie Rocher; Dominique Dardevet

    2014-01-01

    Our aim was to compare and combine 3 nutritional strategies to slow down the age-related loss of muscle mass in healthy old rats: 1) increase protein intake, which is likely to stimulate muscle protein anabolism; 2) use leucine rich, rapidly digested whey proteins as protein source (whey proteins are recognized as the most effective proteins to stimulate muscle protein anabolism). 3) Supplement animals with a mixture of chamomile extract, vitamin E, vitamin D (reducing inflammation and oxidat...

  6. Identification and characterization of the protein components of the skeletal muscle receptor for the 1,4-dihydropyridine Ca2+ channel blockers

    International Nuclear Information System (INIS)

    In these studies, photoaffinity labeling and immunolabeling approaches were used to identify and characterize components of the skeletal muscle receptor for the 1,4-dihydropyridine Ca2+ channel blockers. The 1,4-dihydropyridine receptor purified from rabbit skeletal muscle consists of proteins of 175,000, 170,000, 52,000, and 32,000 Da when analyzed by SDS-PAGE under nonreducing conditions and stained with Coomassie Blue dye. After reduction of disulfide bonds, the 175,000 Da protein shifts in apparent molecular mass to 150,000 Da. Photoaffinity labeling using the dihydropyridine ligands [3H]azidopine and [3H]PN200-110 identified a protein of 170,000 Da as the dihydropyridine binding component of the receptor. Specific polyclonal antibodies were developed against both the nonreduced and reduced forms of the 175/150,000 and 32,000 Da proteins and were used to show that the 150,000 and 32,000 Da proteins are distinct from each other and from other components of the receptor and that they copurify with the 170,000 Da protein at each step of purification. In addition, monoclonal antibodies against the 170,000 and 52,000 Da polypeptides were shown to coimmunoprecipitate the 150,000 and 32,000 Da polypeptides from solubilized skeletal muscle triads

  7. Curvas de crescimento e de deposição dos componentes corporais em aves de postura de diferentes linhagens Growth curves and deposition of body components in pullets of different strains

    Directory of Open Access Journals (Sweden)

    Rafael Neme

    2006-06-01

    and Hisex White (HSW were evaluated. Three hundred birds of each strain were assigned to a completely randomized design with four replicates of 75 birds. Body weight (BW, feather weight (FW and body depositions of protein, fat, ash and water were weekly measured. The growth curves were determined by Gompertz function with data collections. The semi heavy birds were later than the light ones for BW, with greater body growth rates and weights at maturity. The HLW36 were 64.23 g lighter at maturity than the HSW. Data obtained for feather growth and protein deposition were similar, but the semi heavy birds showed greater weight at maturity (WM than the light ones. However, for these components, the birds Hy Line had larger depositions than Hisex. HLW36 strain had smaller feather rate at maturity, making it three days later than HSW. The lowest protein deposition in the end of growth period caused the deviation of energy ingested for fat deposition in HSB birds and overestimated the WM in relation to HLB birds. Hy Line birds had later body ash composition and showed greater WM than Hisex. Among the studied variables, Hy Line birds had always heavier WM, however, the body water content of Hisex birds was greater and the maximum rates at maturity were later. This fact explains the similarity in the weight gain among the studied strains, once other body components were greater for Hy Line birds.

  8. ZNF198, a zinc finger protein rearranged in myeloproliferative disease, localizes to the PML nuclear bodies and interacts with SUMO-1 and PML

    International Nuclear Information System (INIS)

    The ZNF198/FGFR1 fusion gene in atypical myeloproliferative disease produces a constitutively active cytoplasmic tyrosine kinase, unlike ZNF198 which is normally a nuclear protein. We have now shown that the ZNF198/FGFR1 fusion kinase interacts with the endogenous ZNF198 protein suggesting that the function of ZNF198 may be compromised in cells expressing it. Little is currently known about the endogenous function of ZNF198 and to investigate this further we performed a yeast two-hybrid analysis and identified SUMO-1 as a binding partner of ZNF198. These observations were confirmed using co-immunoprecipitation which demonstrated that ZNF198 is covalently modified by SUMO-1. Since many of the SUMO-1-modified proteins are targeted to the PML nuclear bodies we used confocal microscopy to show that SUMO-1, PML and ZNF198 colocalize to punctate structures, shown by immunocytochemistry to be PML bodies. Using co-immunoprecipitation we now show that PML and sumoylated ZNF198 can be found in a protein complex in the cell. Mutation of the SUMO-1 binding site in wild-type ZNF198 resulted in loss of distinct PML bodies, reduced PML levels and a more dispersed nuclear localization of the PML protein. In cells expressing ZNF198/FGFR1, which also lack the SUMO-1 binding site, SUMO-1 is preferentially localized in the cytoplasm, which is associated with loss of distinct PML bodies. Recently, arsenic trioxide (ATO) was proposed as an alternative therapy for APL that was resistant to traditional therapy. Treatment of cells expressing ZNF198/FGFR1 with ATO demonstrated reduced autophosphorylation of the ZNF198/FGFR1 protein and induced apoptosis, which is not seen in cells expressing wild-type ZNF198. Overall our results suggest that the sumoylation of ZNF198 is important for PML body formation and that the abrogation of sumoylation of ZNF198 in ZNF198/FGFR1 expressing cells may be an important mechanism in cellular transformation

  9. Variable stoichiometry of the TatA component of the twin-arginine protein transport system observed by in vivo single-molecule imaging

    OpenAIRE

    Leake, Mark C.; Greene, Nicholas P.; Godun, Rachel M.; Granjon, Thierry; Buchanan, Grant; Chen, Shuyun; Berry, Richard M; Palmer, Tracy; Berks, Ben C.

    2008-01-01

    The twin-arginine translocation (Tat) system transports folded proteins across the bacterial cytoplasmic membrane and the thylakoid membrane of plant chloroplasts. The essential components of the Tat pathway are the membrane proteins TatA, TatB, and TatC. TatA is thought to form the protein translocating element of the Tat system. Current models for Tat transport make predictions about the oligomeric state of TatA and whether, and how, this state changes during the transport cycle. We determi...

  10. Production of soluble truncated spike protein of porcine epidemic diarrhea virus from inclusion bodies of Escherichia coli through refolding.

    Science.gov (United States)

    Piao, Da-Chuan; Lee, Yoon-Seok; Bok, Jin-Duck; Cho, Chong-Su; Hong, Zhong-Shan; Kang, Sang-Kee; Choi, Yun-Jaie

    2016-10-01

    The emergence of highly pathogenic variant porcine epidemic diarrhea virus (PEDV) strains, from 2013 to 2014, in North American and Asian countries have greatly threatened global swine industry. Therefore, development of effective vaccines against PEDV variant strains is urgently needed. Recently, it has been reported that the N-terminal domain (NTD) of S1 domain of PEDV spike protein is responsible for binding to the 5-N-acetylneuraminic acid (Neu5Ac), a possible sugar co-receptor. Therefore, the NTD of S1 domain could be an attractive target for the development of subunit vaccines. In this study, the NTD spanning amino acid residues 25-229 (S25-229) of S1 domain of PEDV variant strain was expressed in Escherichia coli BL21 (DE3) in the form of inclusion bodies (IBs). S25-229 IBs were solubilized in 20 mM sodium acetate (pH 4.5) buffer containing 8 M urea and 1 mM dithiothreitol with 95% yield. Solubilized S25-229 IBs were refolded by 10-fold flash dilution and purified by one-step cation exchange chromatography with >95% purity and 20% yield. The CD spectrum of S25-229 showed the characteristic pattern of alpha helical structure. In an indirect ELISA, purified S25-229 showed strong reactivity with mouse anti-PEDV sera. In addition, immunization of mice with 20 μg of purified S25-229 elicited highly potent serum IgG titers. Finally, mouse antisera against S25-229 showed immune reactivity with native PEDV S protein in an immunofluorescence assay. These results suggest that purified S25-229 may have potential to be used as a subunit vaccine against PEDV variant strains. PMID:27260969

  11. Dcp1-Bodies in Mouse Oocytes

    OpenAIRE

    Swetloff, Adam; Conne, Beatrice; Huarte, Joachim; Pitetti, Jean-Luc; Nef, Serge; Vassalli, Jean-Dominique

    2009-01-01

    Processing bodies (P-bodies) are cytoplasmic granules involved in the storage and degradation of mRNAs. In somatic cells, their formation involves miRNA-mediated mRNA silencing. Many P-body protein components are also found in germ cell granules, such as in mammalian spermatocytes. In fully grown mammalian oocytes, where changes in gene expression depend entirely on translational control, RNA granules have not as yet been characterized. Here we show the presence of P-body-like foci in mouse o...

  12. The multifunctional role of the pallilysin-associated Treponema pallidum protein, Tp0750, in promoting fibrinolysis and extracellular matrix component degradation.

    Science.gov (United States)

    Houston, Simon; Russell, Shannon; Hof, Rebecca; Roberts, Alanna K; Cullen, Paul; Irvine, Kyle; Smith, Derek S; Borchers, Christoph H; Tonkin, Michelle L; Boulanger, Martin J; Cameron, Caroline E

    2014-02-01

    The mechanisms that facilitate dissemination of the highly invasive spirochaete, Treponema pallidum, are incompletely understood. Previous studies showed the treponemal metalloprotease pallilysin (Tp0751) possesses fibrin clot degradation capability, suggesting a role in treponemal dissemination. In the current study we report characterization of the functionally linked protein Tp0750. Structural modelling predicts Tp0750 contains a von Willebrand factor type A (vWFA) domain, a protein-protein interaction domain commonly observed in extracellular matrix (ECM)-binding proteins. We report Tp0750 is a serine protease that degrades the major clot components fibrinogen and fibronectin. We also demonstrate Tp0750 cleaves a matrix metalloprotease (MMP) peptide substrate that is targeted by several MMPs, enzymes central to ECM remodelling. Through proteomic analyses we show Tp0750 binds the endothelial fibrinolytic receptor, annexin A2, in a specific and dose-dependent manner. These results suggest Tp0750 constitutes a multifunctional protein that is able to (1) degrade infection-limiting clots by both inhibiting clot formation through degradation of host coagulation cascade proteins and promoting clot dissolution by complexing with host proteins involved in the fibrinolytic cascade and (2) facilitate ECM degradation via MMP-like proteolysis of host components. We propose that through these activities Tp0750 functions in concert with pallilysin to enable T. pallidum dissemination. PMID:24303899

  13. Immunohistochemical detection of a unique protein within cells of snakes having inclusion body disease, a world-wide disease seen in members of the families Boidae and Pythonidae.

    Directory of Open Access Journals (Sweden)

    Li-Wen Chang

    Full Text Available Inclusion body disease (IBD is a worldwide disease in captive boa constrictors (boa constrictor and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the IBD inclusion bodies, called IBD protein. A validated immuno-based ante-mortem diagnostic test is needed for screening snakes that are at risk of having IBD. In this study, despite difficulties in solubilizing semi-purified inclusion bodies, utilizing hybridoma technology a mouse anti-IBD protein monoclonal antibody (MAB was produced. The antigenic specificity of the antibody was confirmed and validated by western blots, enzyme-linked immunosorbent assay, immuno-transmission electron microscopy, and immunohistochemical staining. Paraffin embedded tissues of IBD positive and negative boa constrictors (n=94 collected from 1990 to 2011 were tested with immunohistochemical staining. In boa constrictors, the anti-IBDP MAB had a sensitivity of 83% and specificity of 100% in detecting IBD. The antibody also cross-reacted with IBD inclusion bodies in carpet pythons (Morelia spilota and a ball python (python regius. This validated antibody can serve as a tool for the development of ante-mortem immunodiagnostic tests for IBD.

  14. Immunohistochemical detection of a unique protein within cells of snakes having inclusion body disease, a world-wide disease seen in members of the families Boidae and Pythonidae.

    Science.gov (United States)

    Chang, Li-Wen; Fu, Ann; Wozniak, Edward; Chow, Marjorie; Duke, Diane G; Green, Linda; Kelley, Karen; Hernandez, Jorge A; Jacobson, Elliott R

    2013-01-01

    Inclusion body disease (IBD) is a worldwide disease in captive boa constrictors (boa constrictor) and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s) and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the IBD inclusion bodies, called IBD protein. A validated immuno-based ante-mortem diagnostic test is needed for screening snakes that are at risk of having IBD. In this study, despite difficulties in solubilizing semi-purified inclusion bodies, utilizing hybridoma technology a mouse anti-IBD protein monoclonal antibody (MAB) was produced. The antigenic specificity of the antibody was confirmed and validated by western blots, enzyme-linked immunosorbent assay, immuno-transmission electron microscopy, and immunohistochemical staining. Paraffin embedded tissues of IBD positive and negative boa constrictors (n=94) collected from 1990 to 2011 were tested with immunohistochemical staining. In boa constrictors, the anti-IBDP MAB had a sensitivity of 83% and specificity of 100% in detecting IBD. The antibody also cross-reacted with IBD inclusion bodies in carpet pythons (Morelia spilota) and a ball python (python regius). This validated antibody can serve as a tool for the development of ante-mortem immunodiagnostic tests for IBD. PMID:24340066

  15. A Fusion between Domains of the Human Bone Morphogenetic Protein-2 and Maize 27 kD γ-Zein Accumulates to High Levels in the Endoplasmic Reticulum without Forming Protein Bodies in Transgenic Tobacco

    Science.gov (United States)

    Ceresoli, Valentina; Mainieri, Davide; Del Fabbro, Massimo; Weinstein, Roberto; Pedrazzini, Emanuela

    2016-01-01

    Human Bone Morphogenetic Protein-2 (hBMP2) is an osteoinductive agent physiologically involved in bone remodeling processes. A commercialized recombinant hBMP2 produced in mammalian cell lines is available in different clinical applications where bone regeneration is needed, but widespread use has been hindered due to an unfavorable cost/effective ratio. Protein bodies are very large insoluble protein polymers that originate within the endoplasmic reticulum by prolamine accumulation during the cereal seed development. The N-terminal domain of the maize prolamin 27 kD γ-zein is able to promote protein body biogenesis when fused to other proteins. To produce high yield of recombinant hBMP2 active domain (ad) in stably transformed tobacco plants we have fused it to the γ-zein domain. We show that this zein-hBMP2ad fusion is retained in the endoplasmic reticulum without forming insoluble protein bodies. The accumulation levels are above 1% of total soluble leaf proteins, indicating that it could be a rapid and suitable strategy to produce hBMP2ad at affordable costs. PMID:27047526

  16. Early Detection of Baculovirus Expression and Infection in Lepidopteran Larvae Fed Occlusion Bodies of an AcMNPV Recombinant Carrying a Red Fluorescent Protein Gene

    Science.gov (United States)

    A method has been devised utilizing a baculovirus recombinant (AcMNPV hsp70Red) carrying a red fluorescent protein (RFP) gene under the early heat shock promoter (hsp70) to assess potential infectivity of larvae fed occlusion bodies. A time study was employed whereby first and third instars of Trich...

  17. Total-body protein turnover in parenterally fed neonates: effects of energy source studied by using [15N]glycine and [1-13C]leucine.

    Science.gov (United States)

    Pencharz, P; Beesley, J; Sauer, P; Van Aerde, J; Canagarayar, U; Renner, J; McVey, M; Wesson, D; Swyer, P

    1989-12-01

    The effects of nonprotein energy source (ie, glucose only vs glucose and lipid) on nitrogen retention and total-body protein turnover were studied in 20 parenterally fed newborn infants. All infants received approximately 3 g amino acids and 80-90 kcal.kg body wt.d. Total-body protein synthesis was estimated by using three constant-infusion, end-product methods: enrichment of urinary urea and ammonia in response to a [15N]glycine label and exhaled carbon dioxide enrichment in response to a [1-13C]leucine label. No differences were seen in nitrogen retention between the two energy sources. The estimate of total-body protein turnover obtained from the 13C label was similar to that obtained with the [15N]urea label. No differences in turnover rates were observed between the two diet groups. Use of the glucose-plus-lipid fuel system enhanced energy storage and the reutilization of amino acid for protein synthesis. PMID:2512806

  18. Accumulation of radium in ferruginous protein bodies formed in lung tissue. Association of resulting radiation hotspots with malignant mesothelioma and other malignancies

    International Nuclear Information System (INIS)

    While exposure to fibers and particles has been proposed to be associated with several different lung malignancies including mesothelioma, the mechanism for the carcinogenesis is not fully understood. Along with mineralogical observation, we have analyzed forty-four major and trace elements in extracted asbestos bodies (fibers and proteins attached to them) with coexisting fiber-free ferruginous protein bodies from extirpative lungs of individuals with malignant mesothelioma. These observations together with patients' characteristics suggest that inhaled iron-rich asbestos fibers and dust particles, and excess iron deposited by continuous cigarette smoking would induce ferruginous protein body formation resulting in ferritin aggregates in lung tissue. Chemical analysis of ferruginous protein bodies extracted from lung tissues reveals anomalously high concentrations of radioactive radium, reaching millions of times higher concentration than that of seawater. Continuous and prolonged internal exposure to hotspot ionizing radiation from radium and its daughter nuclides could cause strong and frequent DNA damage in lung tissue, initiate different types of tumour cells, including malignant mesothelioma cells, and may cause cancers. (author)

  19. Signal-dependent Regulation of Transcription by Histone Deacetylase 7 Involves Recruitment to Promyelocytic Leukemia Protein Nuclear Bodies

    Science.gov (United States)

    Gao, Chengzhuo; Cheng, Xiwen; Lam, Minh; Liu, Yu; Liu, Qing; Chang, Kun-Sang

    2008-01-01

    Promyelocytic leukemia protein (PML) nuclear bodies (NBs) are dynamic subnuclear compartments that play roles in several cellular processes, including apoptosis, transcriptional regulation, and DNA repair. Histone deacetylase (HDAC) 7 is a potent corepressor that inhibits transcription by myocyte enhancer factor 2 (MEF2) transcription factors. We show here that endogenous HDAC7 and PML interact and partially colocalize in PML NBs. Tumor necrosis factor (TNF)-α treatment recruits HDAC7 to PML NBs and enhances association of HDAC7 with PML in human umbilical vein endothelial cells. Consequently, TNF-α promotes dissociation of HDAC7 from MEF2 transcription factors and the promoters of MEF2 target genes such as matrix metalloproteinase (MMP)-10, leading to accumulation of MMP-10 mRNA. Conversely, knockdown of PML enhances the association between HDAC7 and MEF2 and decreases MMP-10 mRNA accumulation. Accordingly, ectopic expression of PML recruits HDAC7 to PML NBs and leads to activation of MEF2 reporter activity. Notably, small interfering RNA knockdown of PML decreases basal and TNF-α-induced MMP-10 mRNA accumulation. Our results reveal a novel mechanism by which PML sequesters HDAC7 to relieve repression and up-regulate gene expression. PMID:18463162

  20. Whey protein improves HDL/non-HDL ratio and body weight gain in rats subjected to the resistance exercise

    Directory of Open Access Journals (Sweden)

    Kely Raspante Teixeira

    2012-12-01

    Full Text Available The aim of this study was to evaluate the effects of resistance exercise, such as weight-lifting (WL on the biochemical parameters of lipid metabolism and cardiovascular disease risk in the rats fed casein (control or whey protein (WP diets. Thirty-two male Fisher rats were randomly assigned to sedentary or exercise-trained groups and were fed control or WP diets. The WL program consisted of inducing the animals to perform the sets of jumps with weights attached to the chest. After seven weeks, arteriovenous blood samples were collected for analysis. The WL or WP ingestion were able to improve the lipid profile, reducing the TC and non-HDL cholesterol concentrations, but only WP treatment significantly increased the serum HDL concentrations, thereby also affecting the TC/HDL and HDL/non-HDL ratios. However, WL plus WP was more effective in improving the HDL/non-HDL ratio than the exercise or WP ingestion alone and the body weight gain than exercise without WP ingestion.

  1. Outer Membrane Targeting of Pseudomonas aeruginosa Proteins Shows Variable Dependence on the Components of Bam and Lol Machineries

    OpenAIRE

    Hoang, Hanh H.; Nickerson, Nicholas N.; Lee, Vincent T.; Kazimirova, Anastasia; Chami, Mohamed; Pugsley, Anthony P.; Lory, Stephen

    2011-01-01

    ABSTRACT In Gram-negative bacteria, the Lol and Bam machineries direct the targeting of lipidated and nonlipidated proteins, respectively, to the outer membrane (OM). Using Pseudomonas aeruginosa strains with depleted levels of specific Bam and Lol proteins, we demonstrated a variable dependence of different OM proteins on these targeting pathways. Reduction in the level of BamA significantly affected the ability of the β-barrel membrane protein OprF to localize to the OM, while the targeting...

  2. Structural and thermodynamic characterization of the interaction between two periplasmic Treponema pallidum lipoproteins that are components of a TPR-protein-associated TRAP transporter (TPAT)

    Science.gov (United States)

    Brautigam, Chad A.; Deka, Ranjit K.; Schuck, Peter; Tomchick, Diana R.; Norgard, Michael V.

    2012-01-01

    Tripartite ATP-independent periplasmic transporters (TRAP-Ts) are bacterial transport systems that have been implicated in the import of small molecules into the cytoplasm. A newly discovered subfamily of TRAP-Ts (TPATs) has four components. Three are common to both TRAP-Ts and TPATs: the P component, a ligand-binding protein, and a transmembrane symporter apparatus comprising the M and Q components (M and Q are sometimes fused to form a single polypeptide). TPATs are distinguished from TRAP-Ts by the presence of a unique protein called the “T component”. In Treponema pallidum, this protein (TatT) is a water-soluble trimer whose protomers are each perforated by a pore. Its respective P component (TatPT) interacts with the TatT in vitro and in vivo. In this work, we further characterized this interaction. Co-crystal structures of two complexes between the two proteins confirm that up to three monomers of TatPT can bind to the TatT trimer. A putative ligand-binding cleft of TatPT aligns with the pore of TatT, strongly suggesting ligand transfer between T and PT. We used a combination of site-directed mutagenesis and analytical ultracentrifugation to derive thermodynamic parameters for the interactions. These observations confirm that the observed crystallographic interface is recapitulated in solution. These results prompt a hypothesis of the molecular mechanism(s) of hydrophobic ligand transport by the TPATs. PMID:22504226

  3. Daily Overfeeding from Protein and/or Carbohydrate Supplementation for Eight Weeks in Conjunction with Resistance Training Does not Improve Body Composition and Muscle Strength or Increase Markers Indicative of Muscle Protein Synthesis and Myogenesis in Resistance-Trained Males.

    Science.gov (United States)

    Spillane, Mike; Willoughby, Darryn S

    2016-03-01

    This study determined the effects of heavy resistance training and daily overfeeding with carbohydrate and/or protein on blood and skeletal muscle markers of protein synthesis (MPS), myogenesis, body composition, and muscle performance. Twenty one resistance-trained males were randomly assigned to either a protein + carbohydrate [HPC (n = 11)] or a carbohydrate [HC (n = 10)] supplement group in a double-blind fashion. Body composition and muscle performance were assessed, and venous blood samples and muscle biopsies were obtained before and after eight weeks of resistance training and supplementation. Data were analyzed by two-way ANOVA (p ≤ 0.05). Total body mass, body water, and fat mass were significantly increased in both groups in response to resistance training, but not supplementation (p 0.05). Muscle total DNA, total protein, and c-Met were not significantly affected (p > 0.05). In conjunction with resistance training, the peri-exercise and daily overfeeding of protein and/or carbohydrate did not preferentially improve body composition, muscle performance, and markers indicative of MPS and myogenic activation. Key pointsIn response to 56 days of heavy resistance training and HC or HPC supplementation, similar increases in muscle mass and strength in both groups occurred; however, the increases were not different between supplement groups.The supplementation of HPC had no preferential effect on augmenting serum IGF-1 GH, or HGF.The supplementation of HPC had no preferential effect on augmenting increases in total muscle protein content or the myogenic markers, total DNA and muscle cMet content.In response to 56 days of a daily supplemental dose of 94 g of protein and 196 g of carbohydrate, the HPC group was no more effective than 312 g of carbohydrate in the HC group in increasing muscle strength and mass due to its ability to elevate serum anabolic hormones and growth factors and markers of myogenic activation of satellite cells. PMID:26957922

  4. Estimation of whole body protein turnover by L-[1-13C]-leucine tracer experiment in post-viral hepatocirrhotic patients

    International Nuclear Information System (INIS)

    A constant infusion of L-[1-1'3C]-leucine given to the subject and the plasma 13C-leucine enrichment and breath 1'3CO2 enrichment were measured with GC-MS and gas isotope ratio MS respectively. The plateau isotope enrichments reached were used to calculate the whole body protein synthesis and breakdown rates using a two pools model. 15 post-viral hepatocirrhotic patients and 6 normal adults were studied. The whole body protein synthesis and breakdown rates were both markedly increased in cirrhotic patients, with the breakdown rate higher than the synthesis rate and hence leading to a negative nitrogen balance. The changes of 7 decompensated cirrhotic patients were more severe than 8 compensated cirrhotic patients. The results suggest that the protein malnutrition and negative nitrogen balance of post-viral hepatocirrhotic patients are not simply due to low intake of protein and that the hyperactive turnover, especially the excessive breakdown of body protein might play an important role

  5. Assessing body condition and energy budget components by scoring abdominal profiles in free-ranging pink-footed geese Anser brachyrhynchus

    NARCIS (Netherlands)

    Madsen, J.; Klaassen, M.R.J.

    2006-01-01

    An abdominal profile index (API) was developed for pink-footed geese Anser brachyrhynchus as a measure of body condition. On basis of carcass analysis of 56 adult geese with known API prior to collection, we found significant linear relationships between API against body mass, abdominal fat and tota

  6. The subcellular distribution of the human ribosomal "stalk" components: P1, P2 and P0 proteins

    DEFF Research Database (Denmark)

    Tchórzewski, Marek; Krokowski, Dawid; Rzeski, Wojciech;

    2003-01-01

    The ribosomal "stalk" structure is a distinct lateral protuberance located on the large ribosomal subunit in prokaryotic, as well as in eukaryotic cells. In eukaryotes, this ribosomal structure is composed of the acidic ribosomal P proteins, forming two hetero-dimers (P1/P2) attached to the ribos......The ribosomal "stalk" structure is a distinct lateral protuberance located on the large ribosomal subunit in prokaryotic, as well as in eukaryotic cells. In eukaryotes, this ribosomal structure is composed of the acidic ribosomal P proteins, forming two hetero-dimers (P1/P2) attached...... to the ribosome through the P0 protein. The "stalk" is essential for the ribosome activity, taking part in the interaction with elongation factors.In this report, we have shown that the subcellular distribution of the human P proteins does not fall into standard behavior of regular ribosomal proteins. We have...... used two approaches to assess the distribution of the P proteins, in vivo experiments with GFP fusion proteins and in vitro one with anti-P protein antibodies. In contrast to standard r-proteins, the P1 and P2 proteins are not actively transported into the nucleus compartment, remaining predominantly...

  7. mRNA decay proteins are targeted to poly(A+ RNA and dsRNA-containing cytoplasmic foci that resemble P-bodies in Entamoeba histolytica.

    Directory of Open Access Journals (Sweden)

    Itzel López-Rosas

    Full Text Available In higher eukaryotes, mRNA degradation and RNA-based gene silencing occur in cytoplasmic foci referred to as processing bodies (P-bodies. In protozoan parasites, the presence of P-bodies and their putative role in mRNA decay have yet to be comprehensively addressed. Identification of P-bodies might provide information on how mRNA degradation machineries evolved in lower eukaryotes. Here, we used immunofluorescence and confocal microscopy assays to investigate the cellular localization of mRNA degradation proteins in the human intestinal parasite Entamoeba histolytica and found evidence of the existence of P-bodies. Two mRNA decay factors, namely the EhXRN2 exoribonuclease and the EhDCP2 decapping enzyme, were localized in cytoplasmic foci in a pattern resembling P-body organization. Given that amoebic foci appear to be smaller and less rounded than those described in higher eukaryotes, we have named them "P-body-like structures". These foci contain additional mRNA degradation factors, including the EhCAF1 deadenylase and the EhAGO2-2 protein involved in RNA interference. Biochemical analysis revealed that EhCAF1 co-immunoprecipitated with EhXRN2 but not with EhDCP2 or EhAGO2-2, thus linking deadenylation to 5'-to-3' mRNA decay. The number of EhCAF1-containing foci significantly decreased after inhibition of transcription and translation with actinomycin D and cycloheximide, respectively. Furthermore, results of RNA-FISH assays showed that (i EhCAF1 colocalized with poly(A(+ RNA and (ii during silencing of the Ehpc4 gene by RNA interference, EhAGO2-2 colocalized with small interfering RNAs in cytoplasmic foci. Our observation of decapping, deadenylation and RNA interference proteins within P-body-like foci suggests that these structures have been conserved after originating in the early evolution of eukaryotic lineages. To the best of our knowledge, this is the first study to report on the localization of mRNA decay proteins within P-body

  8. Interactions of dietary protein and adiposity measures in relation to subsequent changes in body weight and waist circumference

    DEFF Research Database (Denmark)

    Ankarfeldt, Mikkel Z; Angquist, Lars; Jakobsen, Marianne Uhre; Overvad, Kim; Tjønneland, Anne; Halkjaer, Jytte; Astrup, Arne; Sørensen, Thorkild I A

    2014-01-01

    dietary protein, whether replacing carbohydrate or fat, and weight change. However, individuals in the highest tertile of baseline BMI (irrespective of baseline WCBMI ) had significantly inverse change in waist circumference when protein replaced carbohydrate, but not when protein replaced fat. CONCLUSION......: Replacing carbohydrate with protein in the diet may prevent a relative increase in WC in individuals with a greater BMI....

  9. Biosynthesis of intestinal microvillar proteins. Rapid expression of cytoskeletal components in microvilli of pig small intestinal mucosal explants

    DEFF Research Database (Denmark)

    Cowell, G M; Danielsen, E M

    1984-01-01

    Using alkaline extraction to separate cytoskeletal and membrane proteins of intestinal microvilli, the kinetics of assembly of these two microvillar protein compartments was studied by pulse-chase labelling of pig small intestinal mucosal explants, kept in organ culture. Following a 10 min pulse of...... pulse. These different kinetics of appearance indicate that the two microvillar protein compartments have separate mechanisms of biosynthesis and microvillar expression....

  10. Effect of gamma irradiation on nutritional components and Cry1Ab protein in the transgenic rice with a synthetic cry1Ab gene from Bacillus thuringiensis

    International Nuclear Information System (INIS)

    The effects of gamma irradiation on the transgenic rice containing a synthetic cry1Ab gene from Bacillus thuringiensis were investigated. There was almost no difference in the content of the major nutritional components, i.e. crude protein, crude lipid, eight essential amino acids and total ash between the irradiated grains and the non-irradiated transgenic rice. However, the amounts of Cry1Ab protein and apparent amylose in the irradiated transgenic rice were reduced significantly by the doses higher than 200 Gy. In vivo observation showed that Cry1Ab protein contents also decreased in the fresh leaf tissues of survival seedlings after irradiation with 200 Gy or higher doses and showed inhibition of seedling growth. The results indicate that gamma irradiation might improve the quality of transgenic rice due to removal of the toxic Cry1Ab protein

  11. Composition and structure of nucleolar skeleton (nucleolar matrix)——Actin and fibrillarin are two main protein components of nucleolar skeleton

    Institute of Scientific and Technical Information of China (English)

    陈建明; 沈延; 焦仁杰; 翟中和

    1999-01-01

    Purified nucleoli of HeLa cells were treated sequentially with nonionic detergent, nucleic acid enzyme, low salt and high salt. The residual nucleolar structure termed nucleolar skeleton (nucleolar matrix) was shown as a fine network under electron microscope with DGD embedding-unembedding technique. Such structures of BHK-21 cell and mouse liver cell are similar to that of HeLa cell. The protein composition of the nucleolar skeleton of HeLa cells was analyzed. The protein composition of such nucleolar residual shows obvious difference from the compositions of nuclear matrix and chromosome scaffold. The major protein composition of the nucleolar skeleton of HeLa cells contains 6-7 polypeptides. Their molecular weights are about 48, 43, 36 and 33 ku. Further studies show that actin and fib-rillarin are two major protein components of nucleolar skeleton of HeLa cells.

  12. Effect of gamma irradiation on nutritional components and Cry1Ab protein in the transgenic rice with a synthetic cry1Ab gene from Bacillus thuringiensis

    Science.gov (United States)

    Wu, Dianxing; Ye, Qingfu; Wang, Zhonghua; Xia, Yingwu

    2004-01-01

    The effects of gamma irradiation on the transgenic rice containing a synthetic cry1Ab gene from Bacillus thuringiensis were investigated. There was almost no difference in the content of the major nutritional components, i.e. crude protein, crude lipid, eight essential amino acids and total ash between the irradiated grains and the non-irradiated transgenic rice. However, the amounts of Cry1Ab protein and apparent amylose in the irradiated transgenic rice were reduced significantly by the doses higher than 200 Gy. In vivo observation showed that Cry1Ab protein contents also decreased in the fresh leaf tissues of survival seedlings after irradiation with 200 Gy or higher doses and showed inhibition of seedling growth. The results indicate that gamma irradiation might improve the quality of transgenic rice due to removal of the toxic Cry1Ab protein.

  13. Body Mass Index, High-Sensitivity C-Reactive Protein and Mortality in Chinese with Coronary Artery Disease.

    Directory of Open Access Journals (Sweden)

    Ding Ding

    Full Text Available To investigate single and joint associations of body mass index (BMI and serum high-sensitivity C-reactive protein (hsCRP with death.The study included 1871 coronary artery disease (CAD patients aged 40-85 year-old recruited from 2008 to 2011. Cox regression models were used to estimate the association of BMI and hsCRP with mortality. The data was analyzed in 2014.During 3.1 years follow-up, 141 deaths were recorded, 110 died of cardiovascular disease (CVD. After adjustment of major CVD risk factors, there was a J-shaped association between BMI and all-cause and CVD mortality, and a positive association between hsCRP and mortality. The J-shaped association of BMI with mortality was present among patients who never smoked or with elevated hsCRP (≥3.0 mg/L. Compared with overweight (BMI 24-27.9 kg/m2 patients with normal hsCRP (<3.0 mg/L, obese patients (BMI≥28 kg/m2 with elevated hsCRP had a 3.41-fold risk of all-cause mortality (95% CI 1.49-7.80 and a 3.50-fold risk of CVD mortality (1.40-8.75, lean patients (BMI<24 kg/m2 with elevated hsCRP concentration had a 2.54-fold risk of all-cause mortality (1.36-4.74 and a 2.36-fold risk of CVD mortality (1.19-4.70.The association pattern between baseline BMI and mortality changed among different baseline hsCRP concentrations, indicating that low-grade inflammation may be related to BMI and secondary prognosis of CAD.

  14. Feature extraction and fusion for protein structure identification in cryo-electron microscopic images using independent component analysis and the projection-slice synthetic filter

    Science.gov (United States)

    Riasati, Vahid R.; Braunreiter, Dennis

    2006-04-01

    In this paper we utilize the Projection-Slice Synthetic Discriminant Function Filters, PSDF, in concert with an Independent Component Analysis technique to simultaneously reduce the data set that represents each of the training images and to emphasize subtle differences in each of the training images. These differences are encoded into the PSDF in order to improve the filter sensitivity for the recognition and identification of protein images formed from a cryo-electron microscopic imaging process. The PSDF and Independent Component Analysis provide a premise not only for the identification of the class of structures under consideration, but also for detecting the orientation of the structures in these images. The protein structures found in cryo-electron microscopic imaging represent a class of objects that have low resolution and contrast and subtle variation. This poses a challenge in design of filters to recognize these structures due to false targets that often have similar characteristics as the protein structures. The incorporation of a component analysis and eigen values conditioning in forming the filter provides an enhanced approach of de-correlating images prior to their incorporation into the filter. We present our method of filter synthesis and the results of the application of this modified filter to a protein structure recognition problem.

  15. A novel zf-MYND protein, CHB-3, mediates guanylyl cyclase localization to sensory cilia and controls body size of Caenorhabditis elegans.

    Directory of Open Access Journals (Sweden)

    Manabi Fujiwara

    2010-11-01

    Full Text Available Cilia are important sensory organelles, which are thought to be essential regulators of numerous signaling pathways. In Caenorhabditis elegans, defects in sensory cilium formation result in a small-body phenotype, suggesting the role of sensory cilia in body size determination. Previous analyses suggest that lack of normal cilia causes the small-body phenotype through the activation of a signaling pathway which consists of the EGL-4 cGMP-dependent protein kinase and the GCY-12 receptor-type guanylyl cyclase. By genetic suppressor screening of the small-body phenotype of a cilium defective mutant, we identified a chb-3 gene. Genetic analyses placed chb-3 in the same pathway as egl-4 and gcy-12 and upstream of egl-4. chb-3 encodes a novel protein, with a zf-MYND motif and ankyrin repeats, that is highly conserved from worm to human. In chb-3 mutants, GCY-12 guanylyl cyclase visualized by tagged GFP (GCY-12::GFP fails to localize to sensory cilia properly and accumulates in cell bodies. Our analyses suggest that decreased GCY-12 levels in the cilia of chb-3 mutants may cause the suppression of the small-body phenotype of a cilium defective mutant. By observing the transport of GCY-12::GFP particles along the dendrites to the cilia in sensory neurons, we found that the velocities and the frequencies of the particle movement are decreased in chb-3 mutant animals. How membrane proteins are trafficked to cilia has been the focus of extensive studies in vertebrates and invertebrates, although only a few of the relevant proteins have been identified. Our study defines a new regulator, CHB-3, in the trafficking process and also shows the importance of ciliary targeting of the signaling molecule, GCY-12, in sensory-dependent body size regulation in C. elegans. Given that CHB-3 is highly conserved in mammal, a similar system may be used in the trafficking of signaling proteins to the cilia of other species.

  16. Calcium-dependent and -independent binding of the pentraxin serum amyloid P component to glycosaminoglycans and amyloid proteins

    DEFF Research Database (Denmark)

    Danielsen, B; Sørensen, I J; Nybo, Mads; Nielsen, E H; Kaplan, B; Svehag, S E

    1997-01-01

    beta2M) by ELISA. An increase in the dose-dependent binding of SAP to heparan sulfate, AA-protein and beta2M was observed as the pH decreased from 8.0 to 5.0. Furthermore, a lower, but significant Ca2(+)-independent binding of SAP to heparan sulfate, dermatan sulfate, AA protein and the amyloid...

  17. Protection of scaffold protein Isu from degradation by the Lon protease Pim1 as a component of Fe–S cluster biogenesis regulation

    Science.gov (United States)

    Ciesielski, Szymon J.; Schilke, Brenda; Marszalek, Jaroslaw; Craig, Elizabeth A.

    2016-01-01

    Iron–sulfur (Fe–S) clusters, essential protein cofactors, are assembled on the mitochondrial scaffold protein Isu and then transferred to recipient proteins via a multistep process in which Isu interacts sequentially with multiple protein factors. This pathway is in part regulated posttranslationally by modulation of the degradation of Isu, whose abundance increases >10-fold upon perturbation of the biogenesis process. We tested a model in which direct interaction with protein partners protects Isu from degradation by the mitochondrial Lon-type protease. Using purified components, we demonstrated that Isu is indeed a substrate of the Lon-type protease and that it is protected from degradation by Nfs1, the sulfur donor for Fe–S cluster assembly, as well as by Jac1, the J-protein Hsp70 cochaperone that functions in cluster transfer from Isu. Nfs1 and Jac1 variants known to be defective in interaction with Isu were also defective in protecting Isu from degradation. Furthermore, overproduction of Jac1 protected Isu from degradation in vivo, as did Nfs1. Taken together, our results lead to a model of dynamic interplay between a protease and protein factors throughout the Fe–S cluster assembly and transfer process, leading to up-regulation of Isu levels under conditions when Fe–S cluster biogenesis does not meet cellular demands. PMID:26842892

  18. Purification of an 80,000-dalton protein that is a component of the isolated microvillus cytoskeleton, and its localization in nonmuscle cells

    OpenAIRE

    1983-01-01

    The microvillus cytoskeleton, isolated from chicken intestinal epithelial cell brush borders, is known to contain five major protein components, the 110,000-dalton polypeptide, villin (95,000 daltons), fimbrin (68,000 daltons), actin (43,000 daltons), and calmodulin (17,000 daltons). In this paper we describe our first step in studying the minor components of the isolated core. We have so far identified and purified an 80,000-dalton polypeptide that was present in the isolated structure in ap...

  19. Protein-chemical characterization of NF-H, the largest mammalian neurofilament component; intermediate filament-type sequences followed by a unique carboxy-terminal extension

    OpenAIRE

    Geisler, N; Fischer, S.; Vandekerckhove, J; Van Damme, J; Plessmann, U.; Weber, K.

    1985-01-01

    NF-H has the highest mol. wt. of the three mammalian neurofilament components (NF-L, NF-M, NF-H). In spite of its unusually large mol. wt., estimated to be 200 K by gel electrophoresis, NF-H contains sequences which identify it as an integral intermediate filament (IF) protein in its amino-terminal region. We have isolated and partially characterized a basic, non-α-helical segment located at the amino-terminal end with properties similar to headpieces of other non-epithelial IF proteins. The ...

  20. Gorduras de descarte e componentes externos do corpo de novilhos e vacas de descarte de diferentes grupos genéticos Trim fat and external components of the body of steers and cull cows from different genetic groups

    Directory of Open Access Journals (Sweden)

    Jonatan Cattelan

    2010-12-01

    cows, respectively. The animals were maintained at feedlot finished until they reached the finishing degree for slaughter. The experimental diet had a roughage concentrate ratio of 40:60 (dry matter basis, containing 14.2% of crude protein and 2,869kcal of digestible energy kg-1 dry matter. The experimental design was complete randomized in a 2x2 (two categories and two genetic groups factorial scheme. The slaughter weight (PAB (509.67 versus 414.50kg and the empty body weight (PCVZ (433.01 versus 355.27kg, respectively were higher for cows in relation to steers. When the components were expreseds in absolutes values and adjusted for PCVZ and PAB, cull cows showed higher inguinal fat, renal fat and internal total fat weights. When the components weights were adjusted for PAB e PCVZ, the steers showed higher head, foots, tail broom and total external components. The 5/8Ch 3/8Ne animals showed higher head absolute weight and ruminal fat in relation to PCVZ, while the 5/8Ne 3/8Ch showed higher rawhide weight in relation to PAB and PCVZ, and in relation to PAB higher total extern components weight. The carcass dressing percentage and non integrants body parts of carcass didn't show any correlation.

  1. The expression of a xylanase targeted to ER-protein bodies provides a simple strategy to produce active insoluble enzyme polymers in tobacco plants.

    Directory of Open Access Journals (Sweden)

    Immaculada Llop-Tous

    Full Text Available BACKGROUND: Xylanases deserve particular attention due to their potential application in the feed, pulp bleaching and paper industries. We have developed here an efficient system for the production of an active xylanase in tobacco plants fused to a proline-rich domain (Zera of the maize storage protein γ-zein. Zera is a self-assembling domain able to form protein aggregates in vivo packed in newly formed endoplasmic reticulum-derived organelles known as protein bodies (PBs. METHODOLOGY/PRINCIPAL FINDINGS: Tobacco leaves were transiently transformed with a binary vector containing the Zera-xylanase coding region, which was optimized for plant expression, under the control of the 35S CaMV promoter. The fusion protein was efficiently expressed and stored in dense PBs, resulting in yields of up to 9% of total protein. Zera-xylanase was post-translationally modified with high-mannose-type glycans. Xylanase fused to Zera was biologically active not only when solubilized from PBs but also in its insoluble form. The resistance of insoluble Zera-xylanase to trypsin digestion demonstrated that the correct folding of xylanase in PBs was not impaired by Zera oligomerization. The activity of insoluble Zera-xylanase was enhanced when substrate accessibility was facilitated by physical treatments such as ultrasound. Moreover, we found that the thermostability of the enzyme was improved when Zera was fused to the C-terminus of xylanase. CONCLUSION/SIGNIFICANCE: In the present work we have successfully produced an active insoluble aggregate of xylanase fused to Zera in plants. Zera-xylanase chimeric protein accumulates within ER-derived protein bodies as active aggregates that can easily be recovered by a simple density-based downstream process. The production of insoluble active Zera-xylanase protein in tobacco outlines the potential of Zera as a fusion partner for producing enzymes of biotechnological relevance. Zera-PBs could thus become efficient and low

  2. Using 15N-glycine trace technique to observe the influence of growth hormone on the rate of whole body protein metabolism in dwarfism

    International Nuclear Information System (INIS)

    The changes of whole body protein metabolism was studied using one oral dose 15N-glycine. Urine 15N excretion 24 hours before and after injected growth hormone (GH) was measured in 9 cases of dwarfism. The results showed that in children with growth hormone deficiency (CHD) with low nitrogen balance, rate of protein syntheses and 15N retention capability were significantly increased after treatment with GH. Children with constitutional growth delay (CGD) with normal parameters of nitrogen balance showed no difference before and after treatment of GH. It was concluded that 15N trace might be a supplementary tool for the diagnosis and differential diagnosis of dwarfism

  3. Effect of high-level fish meal replacement by plant proteins in gilthead sea bream (Sparus aurata) on growth and body/fillet quality traits

    OpenAIRE

    M. de Francesco; Parisi, G.; Perez Sanchez, J.; Gomez Requeni, P; Medale, Francoise; Kaushik, Sadasivam; M. Mecatti; Poli, B

    2007-01-01

    Juvenile gilthead sea bream (initial body weight ca. 100 g) were reared in an indoor flow through marine water system for 1 year. Fish were fed two isoenergetic [19.2 kJ g−1 dry matter (DM)] and isoproteic (426 g kg−1 DM) diets either based on fish meal (diet FM) or on a mixture of plant protein sources (diet PP), replacing 75% of fish meal protein. The growth trial was conducted in duplicate, two tanks for each dietary treatment. Growth performance and feed utilization were regis...

  4. The effects of consuming a high protein diet (4.4 g/kg/d) on body composition in resistance-trained individuals

    OpenAIRE

    Antonio, Jose; Peacock, Corey A; Ellerbroek, Anya; Fromhoff, Brandon; Silver, Tobin

    2014-01-01

    Background The consumption of dietary protein is important for resistance-trained individuals. It has been posited that intakes of 1.4 to 2.0 g/kg/day are needed for physically active individuals. Thus, the purpose of this investigation was to determine the effects of a very high protein diet (4.4 g/kg/d) on body composition in resistance-trained men and women. Methods Thirty healthy resistance-trained individuals participated in this study (mean ± SD; age: 24.1 ± 5.6 yr; height: 171.4 ± 8.8 ...

  5. Bog bodies

    DEFF Research Database (Denmark)

    Lynnerup, Niels

    2015-01-01

    the bog bodies have been studied using medical and natural scientific methods, and recently many bog bodies have been re-examined using especially modern, medical imaging techniques. Because of the preservation of soft tissue, especially the skin, it has been possible to determine lesions and trauma......In northern Europe during the Iron Age, many corpses were deposited in bogs. The cold, wet and anaerobic environment leads in many cases to the preservation of soft tissues, so that the bodies, when found and excavated several thousand years later, are remarkably intact. Since the 19th century....... Conversely, the preservation of bones is less good, as the mineral component has been leached out by the acidic bog. Together with water-logging of collagenous tissue, this means that if the bog body is simply left to dry out when found, as was the case pre-19th century, the bones may literally warp...

  6. An unbiased nuclear proteomics approach reveals novel nuclear protein components that participates in MAMP-triggered immunity.

    Science.gov (United States)

    Fakih, Zainab; Ahmed, Md Bulbul; Letanneur, Claire; Germain, Hugo

    2016-06-01

    (MAMP)-triggered immunity (MTI) is the first layer of molecular defense encountered by pathogens. Genetic screens have contributed to our knowledge of MTI, but are limited to phenotype-causing mutations. Here we attempt to identify novel factors involved in the early event leading to plant MTI by comparing the nuclear proteomes of two Arabidopsis genotypes treated with chitosan. Our approach revealed that following chitosan treatment, cerk1 plants had many nuclear accumulating proteins in common, but also some unique ones, when compared with Col-0 plants. Analysis of the identified proteins revealed a nuclear accumulation of DNA-modifying enzymes, RNA-binding proteins and ribosomal proteins. Our results demonstrate that nuclear proteomic is a valid, phenotype-independent approach to uncover factor involved in cellular processes. PMID:27177187

  7. Comparative effects of dietary nucleoside-nucleotide mixture and its components on endotoxin induced bacterial translocation and small intestinal injury in protein deficient mice.

    OpenAIRE

    Adjei, A A; Yamauchi, K.; Chan, Y. C.; Konishi, M; Yamamoto, S.

    1996-01-01

    BACKGROUND--Nucleoside-nucleotide mixture has been shown to improve gut morphology and reduce the incidence of bacterial translocation in protein deficient mice. AIMS--To compare the reparative effect of nucleoside-nucleotide mixture and their individual components on maintenance of gut integrity and bacterial translocation based on their differential metabolism and utilisation. METHODS--ICR (CD-1) mice were randomised into eight groups of 10 animals each and fed 20% casein diet (control), pr...

  8. Plasma Membrane-Located Purine Nucleotide Transport Proteins Are Key Components for Host Exploitation by Microsporidian Intracellular Parasites

    OpenAIRE

    Heinz, E; Hacker, C.; Dean, P., Shaukat M.U., Khanna, S.P., Chakraborty, S., Lachab, M., Burnett, A., Davies, A.G., Linfield, E.H.; Mifsud, J.; Goldberg, A. V; Williams, T.A.; Nakjang, S.; Gregory, A.; Hirt, R. P.; Lucocq, J M; Kunji, E.R.S.; Embley, T M

    2014-01-01

    Microsporidia are obligate intracellular parasites of most animal groups including humans, but despite their significant economic and medical importance there are major gaps in our understanding of how they exploit infected host cells. We have investigated the evolution, cellular locations and substrate specificities of a family of nucleotide transport (NTT) proteins from Trachipleistophora hominis, a microsporidian isolated from an HIV/AIDS patient. Transport proteins are critical to microsp...

  9. A novel method for evaluating microglial activation using ionized calcium-binding adaptor protein-1 staining: cell body to cell size ratio

    Directory of Open Access Journals (Sweden)

    Iris Bertha Hovens

    2014-09-01

    Full Text Available Aim: The aim was to validate a newly developed methodology of semi-automatic image analysis to analyze microglial morphology as marker for microglial activation in ionized calcium-binding adaptor protein-1 (IBA-1 stained brain sections. Methods: The novel method was compared to currently used analysis methods, visual characterization of activation stage and optical density measurement, in brain sections of young and aged rats that had undergone surgery or remained naοve. Results: The cell body to cell size ratio of microglia was strongly correlated to the visual characterization activation stage. In addition, we observed specific surgery and age-related changes in cell body size, size of the dendritic processes and cell body to cell size ratio. Conclusion: The novel analysis method provides a sensitive marker for microglial activation in the rat brain, which is quick and easy to perform and provides additional information about microglial morphology.

  10. Depleting components of the THO complex causes increased telomere length by reducing the expression of the telomere-associated protein Rif1p.

    Directory of Open Access Journals (Sweden)

    Tai-Yuan Yu

    Full Text Available Telomere length is regulated mostly by proteins directly associated with telomeres. However, genome-wide analysis of Saccharomyces cerevisiae mutants has revealed that deletion of Hpr1p, a component of the THO complex, also affects telomere length. The THO complex comprises four protein subunits, namely, Tho2p, Hpr1p, Mft1p, and Thp2p. These subunits interplay between transcription elongation and co-transcriptional assembly of export-competent mRNPs. Here we found that the deletion of tho2 or hpr1 caused telomere lengthening by ∼50-100 bps, whereas that of mft1 or thp2 did not affect telomere length. Since the THO complex functions in transcription elongation, we analyzed the expression of telomere-associated proteins in mutants depleted of complex components. We found that both the mRNA and protein levels of RIF1 were decreased in tho2 and hpr1 cells. RIF1 encodes a 1917-amino acid polypeptide that is involved in regulating telomere length and the formation of telomeric heterochromatin. Hpr1p and Tho2p appeared to affect telomeres through Rif1p, as increased Rif1p levels suppressed the telomere lengthening in tho2 and hpr1 cells. Moreover, yeast cells carrying rif1 tho2 or rif1 hpr1 double mutations showed telomere lengths and telomere silencing effects similar to those observed in the rif1 mutant. Thus, we conclude that mutations of components of the THO complex affect telomere functions by reducing the expression of a telomere-associated protein, Rif1p.

  11. Ovary growth and protein levels in ovary and fat body during adult-wintering period in the red mason bee, Osmia rufa

    OpenAIRE

    Wasielewski, Oskar; Giejdasz, Karol; Wojciechowicz, Tatiana; Skrzypski, Marek

    2011-01-01

    Winter months are supposed to be a crucial period for the development of the reproductive system, especially ovaries, in Osmia rufa females. During 7 months of the wintering period (from September till March), the length and width of terminal oocytes and protein concentrations in ovary and fat body tissue extracts of red mason bee females, O. rufa, were studied. Our observations indicate statistically significant differences in the size of terminal oocytes for three different periods of over-...

  12. Comparison of Cell Viability and Embryoid Body Size of Two Embryonic Stem Cell Lines After Different Exposure Times to Bone Morphogenetic Protein 4

    OpenAIRE

    Nehleh Zarei Fard; Tahereh Talaei-Khozani; Soghra Bahmanpour; Tahereh Esmaeilpour

    2015-01-01

    Background: Activation of bone morphogenetic protein 4 (BMP4) signaling pathway in embryonic stem (ES) cells plays an important role in controlling cell proliferation, differentiation, and apoptosis. Adverse effects of BMP4 occur in a time dependent manner; however, little is known about the effect of different time exposure of this growth factor on cell number in culture media. In this study, we investigated the role of two different exposure times to BMP4 in cell viability, embryoid body (E...

  13. Crystallization and preliminary X-ray analysis of FliJ, a cytoplasmic component of the flagellar type III protein-export apparatus from Salmonella sp

    International Nuclear Information System (INIS)

    FliJ is one of the essential cytoplasmic components of the flagellar type III protein-export apparatus; it has been expressed, produced and crystallized and the crystals have been characterized by X-ray diffraction. The axial component proteins of the bacterial flagellum are synthesized in the cytoplasm and then translocated into the central channel of the flagellum by the flagellar type III protein-export apparatus for self-assembly at the distal growing end of the flagellum. FliJ is an essential cytoplasmic component of the export apparatus. In this study, Salmonella FliJ with an extra three residues (glycine, serine and histidine) attached to the N-terminus as the remainder of a His tag (GSH-FliJ) was purified and crystallized. Crystals were obtained by the sitting-drop vapour-diffusion technique using PEG 300 as a precipitant. GSH-FliJ crystals grew in the hexagonal space group P6122 or P6522. While the native crystals diffracted to 3.3 Å resolution, the diffraction resolution limit of mercury derivatives was extended to 2.1 Å. Anomalous and isomorphous difference Patterson maps of the mercury-derivative crystal showed significant peaks in their Harker sections, indicating the usefulness of the derivative data for structure determination

  14. OFD1 and flotillins are integral components of a ciliary signaling protein complex organized by polycystins in renal epithelia and odontoblasts.

    Directory of Open Access Journals (Sweden)

    Stephanie Jerman

    Full Text Available Mutation of the X-linked oral-facial-digital syndrome type 1 (OFD1 gene is embryonic lethal in males and results in craniofacial malformations and adult onset polycystic kidney disease in females. While the OFD1 protein localizes to centriolar satellites, centrosomes and basal bodies, its cellular function and how it relates to cystic kidney disease is largely unknown. Here, we demonstrate that OFD1 is assembled into a protein complex that is localized to the primary cilium and contains the epidermal growth factor receptor (EGFR and domain organizing flotillin proteins. This protein complex, which has similarity to a basolateral adhesion domain formed during cell polarization, also contains the polycystin proteins that when mutant cause autosomal dominant polycystic kidney disease (ADPKD. Importantly, in human ADPKD cells where mutant polycystin-1 fails to localize to cilia, there is a concomitant loss of localization of polycystin-2, OFD1, EGFR and flotillin-1 to cilia. Together, these data suggest that polycystins are necessary for assembly of a novel flotillin-containing ciliary signaling complex and provide a molecular rationale for the common renal pathologies caused by OFD1 and PKD mutations.

  15. Tube and sheet hydro-formed steel components for auto-body application; Composants hydroformes a partir de tubes et de toles pour applications en carrosserie automobile

    Energy Technology Data Exchange (ETDEWEB)

    Drewes, E.J.; Blumel, K.; Lenze, F.J. [Thyssen Krupp Stahl AG (Germany)

    2001-10-01

    In view of the weight reduction strategies required and increasingly implemented by the automobile industry, growing importance is being attached not only to the materials but also the forming methods used. Forming processes using hydraulic media in particular offer the potential to produce more complex parts economically while optimizing component properties. The tube hydroforming (IHU) and hydro-mechanical sheet forming (AHU) processes are ideally suited for the production of automotive components in line with these requirements. (authors)

  16. Evaluation of mass spectrometric data using principal component analysis for determination of the effects of organic lakes on protein binder identification.

    Science.gov (United States)

    Hrdlickova Kuckova, Stepanka; Rambouskova, Gabriela; Hynek, Radovan; Cejnar, Pavel; Oltrogge, Doris; Fuchs, Robert

    2015-11-01

    Matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry is commonly used for the identification of proteinaceous binders and their mixtures in artworks. The determination of protein binders is based on a comparison between the m/z values of tryptic peptides in the unknown sample and a reference one (egg, casein, animal glues etc.), but this method has greater potential to study changes due to ageing and the influence of organic/inorganic components on protein identification. However, it is necessary to then carry out statistical evaluation on the obtained data. Before now, it has been complicated to routinely convert the mass spectrometric data into a statistical programme, to extract and match the appropriate peaks. Only several 'homemade' computer programmes without user-friendly interfaces are available for these purposes. In this paper, we would like to present our completely new, publically available, non-commercial software, ms-alone and multiMS-toolbox, for principal component analyses of MALDI-TOF MS data for R software, and their application to the study of the influence of heterogeneous matrices (organic lakes) for protein identification. Using this new software, we determined the main factors that influence the protein analyses of artificially aged model mixtures of organic lakes and fish glue, prepared according to historical recipes that were used for book illumination, using MALDI-TOF peptide mass mapping. PMID:26505772

  17. Tumour necrosis factor and interleukin-6 production induced by components associated with merozoite proteins of Plasmodium falciparum

    DEFF Research Database (Denmark)

    Jakobsen, P H; Moon, R; Ridley, R G;

    1993-01-01

    of infected erythrocytes. These results indicate that the RAP-1 and MSP-1 proteins themselves do not stimulate the production of TNF. Instead, other components associating with these exoantigens may be responsible for the TNF production. Mouse antisera blocking TNF production stimulated by P. yoelii......P. falciparum merozoite antigens, merozoite surface protein-1 (MSP-1) and rhoptry associated protein-1 (RAP-1), were shown to be liberated into the supernatant of in vitro parasite cultures and to be included in the endotoxin-like exoantigen complex, previously designated Ag7. Material affinity...... purified from culture supernatants, using immobilized monoclonal antibodies specific for RAP-1 or MSP-1, stimulated normal human mononuclear cells to produce TNF and IL-6 in vitro. However, stimulation of TNF was absent, and that of IL-6 was reduced, when the antigens were purified from detergent extracts...

  18. REVIEW: Development of methods for body composition studies

    Science.gov (United States)

    Mattsson, Sören; Thomas, Brian J.

    2006-07-01

    This review is focused on experimental methods for determination of the composition of the human body, its organs and tissues. It summarizes the development and current status of fat determinations from body density, total body water determinations through the dilution technique, whole and partial body potassium measurements for body cell mass estimates, in vivo neutron activation analysis for body protein measurements, dual-energy absorptiometry (DEXA), computed tomography (CT) and magnetic resonance imaging (MRI, fMRI) and spectroscopy (MRS) for body composition studies on tissue and organ levels, as well as single- and multiple-frequency bioimpedance (BIA) and anthropometry as simple easily available methods. Methods for trace element analysis in vivo are also described. Using this wide range of measurement methods, together with gradually improved body composition models, it is now possible to quantify a number of body components and follow their changes in health and disease.

  19. Structural analysis of substrate binding by the TatBC component of the twin-arginine protein transport system.

    Science.gov (United States)

    Tarry, Michael J; Schäfer, Eva; Chen, Shuyun; Buchanan, Grant; Greene, Nicholas P; Lea, Susan M; Palmer, Tracy; Saibil, Helen R; Berks, Ben C

    2009-08-11

    The Tat system transports folded proteins across the bacterial cytoplasmic membrane and the thylakoid membrane of plant chloroplasts. In Escherichia coli substrate proteins initially bind to the integral membrane TatBC complex which then recruits the protein TatA to effect translocation. Overproduction of TatBC and the substrate protein SufI in the absence of TatA led to the accumulation of TatBC-SufI complexes that could be purified using an affinity tag on the substrate. Three-dimensional structures of the TatBC-SufI complexes and unliganded TatBC were obtained by single-particle electron microscopy and random conical tilt reconstruction. Comparison of the structures shows that substrate molecules bind on the periphery of the TatBC complex and that substrate binding causes a significant reduction in diameter of the TatBC part of the complex. Although the TatBC complex contains multiple copies of the signal peptide-binding TatC protomer, purified TatBC-SufI complexes contain only 1 or 2 SufI molecules. Where 2 substrates are present in the TatBC-SufI complex, they are bound at adjacent sites. These observations imply that only certain TatC protomers within the complex interact with substrate or that there is a negative cooperativity of substrate binding. Similar TatBC-substrate complexes can be generated by an alternative in vitro reconstitution method and using a different substrate protein. PMID:19666509

  20. A component of the Sec61 ER protein transporting pore is required for plant susceptibility to powdery mildew

    DEFF Research Database (Denmark)

    Zhang, Wen-Jing; Hanisch, Susanne; Kwaaitaal, Mark Adrianus Cornelis J;

    2013-01-01

    EHM, the fungus obtains nutrients from and secretes effector proteins into the plant cell. In the plant cell these effectors interfere with cellular processes such as pathogen defense and membrane trafficking. However, the mechanisms behind effector delivery are largely unknown. This paper provides a...... retrotranslocon pores, the latter being part of the ER-associated protein degradation machinery. We provide support for a model suggesting that the retrotranslocon function of HvSec61βa is required for successful powdery mildew fungal infection. HvSec61βa-GFP and a luminal ER marker were co-localized to the ER....... Effector transport across this EHM-ER interface may occur by a vesicle-mediated process, while the Sec61 retrotranslocon pore potentially provides an escape route for these proteins to reach the cytosol....

  1. Recombinant toluene-4-monooxygenase: catalytic and Mössbauer studies of the purified diiron and rieske components of a four-protein complex.

    Science.gov (United States)

    Pikus, J D; Studts, J M; Achim, C; Kauffmann, K E; Münck, E; Steffan, R J; McClay, K; Fox, B G

    1996-07-16

    Expression of the tmoA-F gene cluster from Pseudomonas mendocina KRI in Escherichia coli BL21(DE3) produces a catalytically active form of the toluene-4-monooxygenase (T4MO) complex. Here we report the purification and characterization of four soluble proteins required for the in vitro reconstitution of T4MO catalytic activity. These proteins are a diiron hydroxylase (T4MOH), a Riesketype ferredoxin (T4MOC), an effector protein (T4MOD), and an NADH oxidoreductase (T4MOF). The T4MOH component is composed of the tmoA, tmoB, and tmoE gene products [quaternary structure (alpha beta epsilon)2, Mr approximately 220 kDa]. The T4MOA polypeptide contains two copies of the amino acid sequence motif (D/E)X(28-37)DEXRH; the same motif provides all of the protein-derived ligands to the diiron centers of ribonucleotide reductase, the soluble methane monooxygenase, and the stearoyl-ACP delta 9 desaturase. Mössbauer, optical, and EPR measurements show that the T4MOH contains diiron centers and suggest that the diiron center contains hydroxo bridge(s) in the diferric state, as observed for methane monooxygenase. Mössbauer and EPR measurements also show that the T4MOC contains a Rieske-type iron-sulfur center. This assignment is in accord with the presence of the amino acid sequence motif CPHX(15-17)CX2H, which has also been found in the bacterial, chloroplastic, and mitochondrial Rieske proteins as well as the bacterial NADH-dependent cis-dihydrodiol-forming aromatic dioxygenases. While single-turnover catalytic studies confirm the function of the T4MOH as the hydroxylase, the NADH-dependent multiple-turnover hydroxylation activity is increased by more than 100-fold in the presence of the T4MOC, which mediates highly specific electron transfer between the T4MOF and the T4MOH. The T4MOD can be purified as an 11.6 kDa monomeric protein devoid of cofactors or redox-active metal ions; this component is also detected as a substoichiometric consitutent of the purified T4MOH. The rate

  2. Human TREX2 components PCID2 and centrin 2, but not ENY2, have distinct functions in protein export and co-localize to the centrosome

    International Nuclear Information System (INIS)

    TREX-2 is a five protein complex, conserved from yeast to humans, involved in linking mRNA transcription and export. The centrin 2 subunit of TREX-2 is also a component of the centrosome and is additionally involved in a distinctly different process of nuclear protein export. While centrin 2 is a known multifunctional protein, the roles of other human TREX-2 complex proteins other than mRNA export are not known. In this study, we found that human TREX-2 member PCID2 but not ENY2 is involved in some of the same cellular processes as those of centrin 2 apart from the classical TREX-2 function. PCID2 is present at the centrosome in a subset of HeLa cells and this localization is centrin 2 dependent. Furthermore, the presence of PCID2 at the centrosome is prevalent throughout the cell cycle as determined by co-staining with cyclins E, A and B. PCID2 but not ENY2 is also involved in protein export. Surprisingly, siRNA knockdown of PCID2 delayed the rate of nuclear protein export, a mechanism distinct from the effects of centrin 2, which when knocked down inhibits export. Finally we showed that co-depletion of centrin 2 and PCID2 leads to blocking rather than delaying nuclear protein export, indicating the dominance of the centrin 2 phenotype. Together these results represent the first discovery of specific novel functions for PCID2 other than mRNA export and suggest that components of the TREX-2 complex serve alternative shared roles in the regulation of nuclear transport and cell cycle progression. - Highlights: • TREX2 complex member PCID2 but not ENY2 localizes to the centrosome in HeLa cells. • Centrin 2 is required for the localization of PCID2 at the centrosome. • PCID2 is found at the centrosome in G1/S at a slightly higher rate than that in G2/M. • PCID2 but not ENY2 delays the rate of nuclear protein export. • Co-depletion of centrin 2 and PCID2 leads to blocking rather than delaying nuclear protein export

  3. Differences in two-component signal transduction proteins among the genus Brucella: implications for host preference and pathogenesis

    DEFF Research Database (Denmark)

    Binnewies, Tim Terence; Ussery, David; Lavín, JL;

    2010-01-01

    . anthropi lacks orthologs of the Brucella TCSs NodVW, TceSR and TcfSR, suggesting that these TCS proteins could be necessary for the adaptation of Brucella as an intracellular pathogen. This genomic analysis revealed the presence of a differential distribution of TCS pseudogenes among Brucella species...

  4. Spectral components of human cardiovascular responses to step changes in Lower Body Negative Pressure (LBNP) before and after 22 hour of 6 deg head down bed rest

    Science.gov (United States)

    Knapp, C. F.; Evans, J. M.; Grande, K. J.; Murphy, C. D.; Patwardhan, A. R.

    1992-01-01

    Changes in autonomic outflow to peripheral organs during the development of bedrest induced orthostatic intolerance have not been determined. Recent studies have indicated that spectral analysis provides an indirect assessment of these changes. Eight male subjects were studied before and after 22 hours of 6 degree head down bedrest plus Lasix (40 mg. P.P.). Cardiovascular spectra (using an autoregressive technique) were determined for heart rate (HR, ECG), arterial pressure (AP, Finapres), radial artery flow (RF, Hokansen) and respiration rate (RR, BoMed). Spectra were obtained from 2.5 minute segments during control, lower body negative pressure (minus 10, 20, 30, 40, 50 mmHg) and recovery. Bedrest increased HR spectra power in the low frequency (.001 to .041 Hz) range, increased RF power in the low and mid (.04 to .18 Hz) range and increased AP power in the high (.18 to .50 Hz) frequency range. Increasing levels of lower body negative pressure decreased HR power and increased RF power in the high frequency range and decreased AP power in the low frequency range. Since spectral power of HR in the high frequency range has been shown to indicate parasympathetically mediated regulation and power in the low and mid frequency ranges indicates a sympathetic / parasympathetic mixture, then both bedrest and lower body negative pressure appeared to shift sympathetic / parasympathetic balance toward sympathetic regulation of HR. The interpretation of the spectral content of AP and RF with respect to their autonomic origins remains unclear.

  5. Plasma membrane-located purine nucleotide transport proteins are key components for host exploitation by microsporidian intracellular parasites.

    Directory of Open Access Journals (Sweden)

    Eva Heinz

    2014-12-01

    Full Text Available Microsporidia are obligate intracellular parasites of most animal groups including humans, but despite their significant economic and medical importance there are major gaps in our understanding of how they exploit infected host cells. We have investigated the evolution, cellular locations and substrate specificities of a family of nucleotide transport (NTT proteins from Trachipleistophora hominis, a microsporidian isolated from an HIV/AIDS patient. Transport proteins are critical to microsporidian success because they compensate for the dramatic loss of metabolic pathways that is a hallmark of the group. Our data demonstrate that the use of plasma membrane-located nucleotide transport proteins (NTT is a key strategy adopted by microsporidians to exploit host cells. Acquisition of an ancestral transporter gene at the base of the microsporidian radiation was followed by lineage-specific events of gene duplication, which in the case of T. hominis has generated four paralogous NTT transporters. All four T. hominis NTT proteins are located predominantly to the plasma membrane of replicating intracellular cells where they can mediate transport at the host-parasite interface. In contrast to published data for Encephalitozoon cuniculi, we found no evidence for the location for any of the T. hominis NTT transporters to its minimal mitochondria (mitosomes, consistent with lineage-specific differences in transporter and mitosome evolution. All of the T. hominis NTTs transported radiolabelled purine nucleotides (ATP, ADP, GTP and GDP when expressed in Escherichia coli, but did not transport radiolabelled pyrimidine nucleotides. Genome analysis suggests that imported purine nucleotides could be used by T. hominis to make all of the critical purine-based building-blocks for DNA and RNA biosynthesis during parasite intracellular replication, as well as providing essential energy for parasite cellular metabolism and protein synthesis.

  6. The effect of dietary protein on reproduction in the mare. II. Growth of foals, body mass of mares and serum protein concentration of mares during the anovulatory, transitional and pregnant periods

    Directory of Open Access Journals (Sweden)

    F.E. Van Niekerk

    1997-07-01

    Full Text Available The effect of 4 different diets, in terms of protein quantity and quality, on total serum protein (TSP, albumin and globulin was investigated. Non-pregnant mares that were not lactating (n = 36, pregnant mares that had foaled (n = 24 and their foals (n = 24 were used in this study. Daily total protein intake had no effect on blood protein concentrations in the mares. Total protein intake and quality (available essential amino-acids did affect the body mass of mares during lactation. When mares were fed the minimum recommended (National Research Council 1989 total daily protein, foal mass decreased by approximately 25 % at weaning compared to the foals whose dams were on a higher level of protein intake. The TSP concentrations of foals at birth were on average 10 g/ℓ lower than those of the mares. Albumin concentrations of foals during the first 60 days of life were on average 2-3 g/ℓ lower than those of the mares. Globulin concentrations of foals were approximately 5 g/ℓ lower than those of mares at weaning.

  7. Theoretical analysis of the geometrical isotope effect on the hydrogen bonds in photoactive yellow protein with multi-component density functional theory

    International Nuclear Information System (INIS)

    Highlights: • We analyze nuclear quantum effects on hydrogen bonds around the chromophore in the photoactive yellow protein. • Multi-component density functional theory is used to analyze the H/D isotope effect on hydrogen bond distances. • The donor–acceptor distance in the hydrogen bonds are elongated by the deuterium substitution. - Abstract: To theoretically analyze the nuclear quantum effects of protons on two hydrogen bonds around the chromophore (CRO) in the photoactive yellow protein (PYP), we have calculated simple cluster model consisting of CRO, Glu46, and Tyr42 residues in PYP with the multi-component molecular orbital method and multi-component density functional theory, which can take account of quantum fluctuations of light mass particles. The average OO distances between CRO and Glu46 and between CRO and Tyr42 with our methods are shorter than the corresponding equilibrium ones, while the OH distances become longer due to the anharmonicity of the potential. The H/D geometrical isotope effect is also found, that is, the distances between oxygen atoms are elongated by the deuterium substitution, known as Ubbelohde effect

  8. Regular multi-component exercise increases physical fitness and muscle protein anabolism in frail, obese, older adults

    OpenAIRE

    Villareal, Dennis T.; Smith, Gordon I.; Sinacore, David R.; Shah, Krupa; Mittendorfer, Bettina

    2010-01-01

    Aging is associated with a decline in strength, endurance, balance, and mobility. Obesity worsens the age-related impairment in physical function and often leads to frailty. The American College of Sports Medicine recommends a multi-component (strength, endurance, flexibility, and balance) exercise program to maintain physical fitness. However, the effect of such an exercise program on physical fitness in frail, obese older adults is not known. We therefore determined the effect of a 3 month-...

  9. Cingulin, a specific protein component of tight junctions, is expressed in normal and neoplastic human epithelial tissues.

    OpenAIRE

    Citi, S.; Amorosi, A; Franconi, F.; Giotti, A.; Zampi, G.

    1991-01-01

    Cingulin is a 140-kd protein localized on the cytoplasmic face of avian tight junctions. The expression of cingulin in human normal and neoplastic colonic tissue has been investigated with an antiserum against chicken cingulin. Human cingulin shares its apparent molecular mass and localization with avian cingulin. In normal colonic epithelium, villous adenomas, and differentiated adenocarcinomas, cingulin staining is observed in the junctional region of the polarized cells lining the surface,...

  10. Ku autoantigen is the regulatory component of a template-associated protein kinase that phosphorylates RNA polymerase II.

    OpenAIRE

    Dvir, A; Peterson, S R; Knuth, M W; Lu, H.; Dynan, W S

    1992-01-01

    The carboxyl-terminal domain of RNA polymerase II contains a tandemly repeated heptapeptide sequence. Previous work has shown that this sequence is phosphorylated at multiple sites by a template-associated protein kinase, in a reaction that is closely associated with the initiation of RNA synthesis. We have purified this kinase to apparent homogeneity from human (HeLa) cells. The purified kinase phosphorylates native RNA polymerase II only in the presence of DNA and the general transcription ...

  11. Phospholipase C-eta enzymes as putative protein kinase C and Ca2+ signalling components in neuronal and neuroendocrine tissues

    OpenAIRE

    Stewart, Alan J.; Morgan, Kevin; Farquharson, Colin; Millar, Robert P

    2007-01-01

    Phosphoinositol-specific phospholipase C enzymes (PLCs) are central to inositol lipid signalling pathways, facilitating intracellular Ca2+ release and protein kinase C activation. A sixth class of phosphoinositol-specific PLC with a novel domain structure, PLC-eta (PLCeta) has recently been discovered in mammals. Recent research, reviewed here, shows that this class consists of two enzymes, PLCeta1 and PLCeta2. Both enzymes hydrolyze phosphatidylinositol 4,5-bisphosphate and are more sensitiv...

  12.  Surfactant Protein D of the Innate Immune Defence is Inversely Associated with Human Obesity and SP-D Deficiency Infers Increased Body Weight in Mice

    DEFF Research Database (Denmark)

    Sørensen, Grith Lykke; Hjelmborg, Jacob v. B.; Leth-Larsen, Rikke;

    2006-01-01

    Surfactant protein D (SP-D) is a key regulator of pathogen-induced inflammation. SP-D is further involved in lipid homeostasis in mouse lung and circulation and recent data have demonstrated that the body mass index (BMI; in kg/m(2)) is influenced by genes in common with SP-D. The objective of th...... males on normal chow. Fat percentage was significantly increased by 17% in the Spd-/- male mice (P = 0.003). We conclude, that there is an association between low levels or absent SP-D and obesity.......Surfactant protein D (SP-D) is a key regulator of pathogen-induced inflammation. SP-D is further involved in lipid homeostasis in mouse lung and circulation and recent data have demonstrated that the body mass index (BMI; in kg/m(2)) is influenced by genes in common with SP-D. The objective of the...... present study was to describe the association between serum SP-D and weight, waist circumference or BMI, and furthermore to observe body weight development in SP-D-deficient (Spd-/-) mice. As a part of the Danish population-based twin study (GEMINAKAR) on the metabolic syndrome, we analysed 1476 Danish...

  13. Surfactant protein D of the innate immune defence is inversely associated with human obesity and SP-D deficiency infers increased body weight in mice

    DEFF Research Database (Denmark)

    Sorensen, G L; Hjelmborg, J V B; Leth-Larsen, R;

    2006-01-01

    Surfactant protein D (SP-D) is a key regulator of pathogen-induced inflammation. SP-D is further involved in lipid homeostasis in mouse lung and circulation and recent data have demonstrated that the body mass index (BMI; in kg/m(2)) is influenced by genes in common with SP-D. The objective of th...... males on normal chow. Fat percentage was significantly increased by 17% in the Spd-/- male mice (P = 0.003). We conclude, that there is an association between low levels or absent SP-D and obesity.......Surfactant protein D (SP-D) is a key regulator of pathogen-induced inflammation. SP-D is further involved in lipid homeostasis in mouse lung and circulation and recent data have demonstrated that the body mass index (BMI; in kg/m(2)) is influenced by genes in common with SP-D. The objective of the...... present study was to describe the association between serum SP-D and weight, waist circumference or BMI, and furthermore to observe body weight development in SP-D-deficient (Spd-/-) mice. As a part of the Danish population-based twin study (GEMINAKAR) on the metabolic syndrome, we analysed 1476 Danish...

  14. Methodical studies on measuring kinetic parameters of the protein metabolism of the whole body in connection with measurings of the energy metabolic rate of rats

    International Nuclear Information System (INIS)

    In 3 successive experiments with growing rats the suitability of pulse labelling with 15N glycine, linked with labelling by means of 14C lysine (experiment 3), was tested for the determination of kinetic parameters of the protein metabolism of the whole body by the application of the compartment model in comparison with pulse labelling with 15N-amino acid mixture (experiment 2) and long-term labelling with 15N-labelled wheat in the feed (experiment 1) under standardized experiment conditions. In measurings of energy metabolism simultaneously carried out with parallel groups of animals the comparability of the metabolic development was studied. The ascertained values of protein synthesis rate, protein catabolism rate and re-utilization rate showed insignificant differences only between the 3 15N tracer variants (with certain limitations for the 'protein turnover' (P)-group of experiment 2) in comparison with errors of the applied methods, from which conclusions can be drawn for the suitability of 15N glycine as tracer, at least under the experimental conditions tested. Protein synthesis and degradation rates ascertained from 14CO2 excretion in experiment 3 were clearly below those average values ascertained with 15N. The differences in the average heat production between the main periods of the 3 experiments were statistically insignificant. (author)

  15. Soldier use of dietary supplements, including protein and body building supplements, in a combat zone is different than use in garrison.

    Science.gov (United States)

    Austin, Krista G; McLellan, Tom M; Farina, Emily K; McGraw, Susan M; Lieberman, Harris R

    2016-01-01

    United States Army personnel in garrison who are not deployed to combat theater report using dietary supplements (DSs) to promote health, increase physical and mental strength, and improve energy levels. Given the substantial physical and cognitive demands of combat, DS use may increase during deployment. This study compared DS use by garrison soldiers with DS use by personnel deployed to a combat theater in Afghanistan. Prevalence and patterns of DS use, demographic factors, and health behaviors were assessed by survey (deployed n = 221; garrison n = 1001). Eighty-two percent of deployed and 74% of garrison soldiers used DSs ≥ 1 time·week(-1). Logistic regression analyses, adjusted for significant demographic and health predictors of DS use, showed deployed personnel were more likely than garrison soldiers to use protein, amino acids, and combination products. Deployed females were more likely to use protein supplements and deployed males were more likely to use multivitamins, combination products, protein, and body building supplements than garrison respondents. Significantly more deployed (17%) than garrison (10%) personnel spent more than $50∙month(-1) on DSs. Higher protein supplement use among deployed personnel was associated with higher frequency of strength training and lower amounts of aerobic exercise for males but similar amounts of strength training and aerobic exercise for females. Protein supplements and combination products are used more frequently by deployed than garrison soldiers with the intent of enhancing strength and energy. PMID:26702674

  16. Time-of-day- and light-dependent expression of ubiquitin protein ligase E3 component N-recognin 4 (UBR4 in the suprachiasmatic nucleus circadian clock.

    Directory of Open Access Journals (Sweden)

    Harrod H Ling

    Full Text Available Circadian rhythms of behavior and physiology are driven by the biological clock that operates endogenously but can also be entrained to the light-dark cycle of the environment. In mammals, the master circadian pacemaker is located in the suprachiasmatic nucleus (SCN, which is composed of individual cellular oscillators that are driven by a set of core clock genes interacting in transcriptional/translational feedback loops. Light signals can trigger molecular events in the SCN that ultimately impact on the phase of expression of core clock genes to reset the master pacemaker. While transcriptional regulation has received much attention in the field of circadian biology in the past, other mechanisms including targeted protein degradation likely contribute to the clock timing and entrainment process. In the present study, proteome-wide screens of the murine SCN led to the identification of ubiquitin protein ligase E3 component N-recognin 4 (UBR4, a novel E3 ubiquitin ligase component of the N-end rule pathway, as a time-of-day-dependent and light-inducible protein. The spatial and temporal expression pattern of UBR4 in the SCN was subsequently characterized by immunofluorescence microscopy. UBR4 is expressed across the entire rostrocaudal extent of the SCN in a time-of-day-dependent fashion. UBR4 is localized exclusively to arginine vasopressin (AVP-expressing neurons of the SCN shell. Upon photic stimulation in the early subjective night, the number of UBR4-expressing cells within the SCN increases. This study is the first to identify a novel E3 ubiquitin ligase component, UBR4, in the murine SCN and to implicate the N-end rule degradation pathway as a potential player in regulating core clock mechanisms and photic entrainment.

  17. Growth performance and body composition of kutum fingerlings, Rutilus frisii kutum (Kamenskii 1901),in response to dietary protein levels

    OpenAIRE

    EBRAHIMI, Ghaffar; OURAJI, Hossein

    2012-01-01

    A feeding trial was conducted to determine the optimal dietary crude protein requirement for Rutilus frisii kutum fingerlings. Five isoenergetic experimental diets were formulated to contain graded levels of protein (32%, 37%, 42%, 47%, and 52%). Triplicate groups of 80 kutum (average weight 0.5 g) were stocked in 250-L tanks and fed to apparent satiation 3 times a day for 8 weeks. The results indicate that the growth performance and feed utilization of kutum were significantly (P < 0.05) ...

  18. Light aerobic physical exercise in combination with leucine and/or glutamine-rich diet can improve the body composition and muscle protein metabolism in young tumor-bearing rats.

    Science.gov (United States)

    Salomão, Emilianne Miguel; Gomes-Marcondes, Maria Cristina Cintra

    2012-12-01

    Nutritional supplementation with some amino acids may influence host's responses and also certain mechanism involved in tumor progression. It is known that exercise influences body weight and muscle composition. Previous findings from our group have shown that leucine has beneficial effects on protein composition in cachectic rat model as the Walker 256 tumor. The main purpose of this study was to analyze the effects of light exercise and leucine and/or glutamine-rich diet in body composition and skeletal muscle protein synthesis and degradation in young tumor-bearing rats. Walker tumor-bearing rats were subjected to light aerobic exercise (swimming 30 min/day) and fed a leucine-rich (3%) and/or glutamine-rich (4%) diet for 10 days and compared to healthy young rats. The carcasses were analyzed as total water and fat body content and lean body mass. The gastrocnemious muscles were isolated and used for determination of total protein synthesis and degradation. The chemical body composition changed with tumor growth, increasing body water and reducing body fat content and total body nitrogen. After tumor growth, the muscle protein metabolism was impaired, showing that the muscle protein synthesis was also reduced and the protein degradation process was increased in the gastrocnemius muscle of exercised rats. Although short-term exercise (10 days) alone did not produce beneficial effects that would reduce tumor damage, host protein metabolism was improved when exercise was combined with a leucine-rich diet. Only total carcass nitrogen and protein were recovered by a glutamine-rich diet. Exercise, in combination with an amino acid-rich diet, in particular, leucine, had effects beyond reducing tumoral weight such as improving protein turnover and carcass nitrogen content in the tumor-bearing host. PMID:22460363

  19. The Alstrom syndrome protein, ALMS1, interacts with α-actinin and components of the endosome recycling pathway.

    Directory of Open Access Journals (Sweden)

    Gayle B Collin

    Full Text Available Alström syndrome (ALMS is a progressive multi-systemic disorder characterized by cone-rod dystrophy, sensorineural hearing loss, childhood obesity, insulin resistance and cardiac, renal, and hepatic dysfunction. The gene responsible for Alström syndrome, ALMS1, is ubiquitously expressed and has multiple splice variants. The protein encoded by this gene has been implicated in ciliary function, cell cycle control, and intracellular transport. To gain better insight into the pathways through which ALMS1 functions, we carried out a yeast two hybrid (Y2H screen in several mouse tissue libraries to identify ALMS1 interacting partners. The majority of proteins found to interact with the murine carboxy-terminal end (19/32 of ALMS1 were α-actinin isoforms. Interestingly, several of the identified ALMS1 interacting partners (α-actinin 1, α-actinin 4, myosin Vb, rad50 interacting 1 and huntingtin associated protein1A have been previously associated with endosome recycling and/or centrosome function. We examined dermal fibroblasts from human subjects bearing a disruption in ALMS1 for defects in the endocytic pathway. Fibroblasts from these patients had a lower uptake of transferrin and reduced clearance of transferrin compared to controls. Antibodies directed against ALMS1 N- and C-terminal epitopes label centrosomes and endosomal structures at the cleavage furrow of dividing MDCK cells, respectively, suggesting isoform-specific cellular functions. Our results suggest a role for ALMS1 variants in the recycling endosome pathway and give us new insights into the pathogenesis of a subset of clinical phenotypes associated with ALMS.

  20. Podoplanin is a component of extracellular vesicles that reprograms cell-derived exosomal proteins and modulates lymphatic vessel formation

    Science.gov (United States)

    Andrés, Germán; Gopal, Shashi K.; Martín-Villar, Ester; Renart, Jaime; Simpson, Richard J.; Quintanilla, Miguel

    2016-01-01

    Podoplanin (PDPN) is a transmembrane glycoprotein that plays crucial roles in embryonic development, the immune response, and malignant progression. Here, we report that cells ectopically or endogenously expressing PDPN release extracellular vesicles (EVs) that contain PDPN mRNA and protein. PDPN incorporates into membrane shed microvesicles (MVs) and endosomal-derived exosomes (EXOs), where it was found to colocalize with the canonical EV marker CD63 by immunoelectron microscopy. We have previously found that expression of PDPN in MDCK cells induces an epithelial-mesenchymal transition (EMT). Proteomic profiling of MDCK-PDPN cells compared to control cells shows that PDPN-induced EMT is associated with upregulation of oncogenic proteins and diminished expression of tumor suppressors. Proteomic analysis of exosomes reveals that MDCK-PDPN EXOs were enriched in protein cargos involved in cell adhesion, cytoskeletal remodeling, signal transduction and, importantly, intracellular trafficking and EV biogenesis. Indeed, expression of PDPN in MDCK cells stimulated both EXO and MV production, while knockdown of endogenous PDPN in human HN5 squamous carcinoma cells reduced EXO production and inhibited tumorigenesis. EXOs released from MDCK-PDPN and control cells both stimulated in vitro angiogenesis, but only EXOs containing PDPN were shown to promote lymphatic vessel formation. This effect was mediated by PDPN on the surface of EXOs, as demonstrated by a neutralizing specific monoclonal antibody. These results contribute to our understanding of PDPN-induced EMT in association to tumor progression, and suggest an important role for PDPN in EV biogenesis and/or release and for PDPN-EXOs in modulating lymphangiogenesis. PMID:26893367

  1. The influence of ionizing radiation in wide dose range on the structural and functional characteristics of protein and lipid components of erythrocyte plasma membranes

    International Nuclear Information System (INIS)

    The mechanisms of low and high dose radiation biological effects is studied by means of erythrocyte ghosts irradiation with doses of 4 x 10-3 - 103 Gy. Changes in the membrane lipid microviscosity, membrane proteins structural mobility, membrane surface potential and intensity of the lipid peroxidation processes were determined. Variations in all investigated parameters take place in ranges of effect as low, so high ionizing radiation doses. Low dose ionizing radiation effects significantly on structural properties of plasma membrane lipid phase, whereas high dose irradiation leads to significant changes in structural parameters of protein component of membranes. It has been established that the features of membrane structural changes are characterized by polyphase changes of examined parameters

  2. Bovine proteins containing poly-glutamine repeats are often polymorphic and enriched for components of transcriptional regulatory complexes

    LENUS (Irish Health Repository)

    Whan, Vicki

    2010-11-23

    Abstract Background About forty human diseases are caused by repeat instability mutations. A distinct subset of these diseases is the result of extreme expansions of polymorphic trinucleotide repeats; typically CAG repeats encoding poly-glutamine (poly-Q) tracts in proteins. Polymorphic repeat length variation is also apparent in human poly-Q encoding genes from normal individuals. As these coding sequence repeats are subject to selection in mammals, it has been suggested that normal variations in some of these typically highly conserved genes are implicated in morphological differences between species and phenotypic variations within species. At present, poly-Q encoding genes in non-human mammalian species are poorly documented, as are their functions and propensities for polymorphic variation. Results The current investigation identified 178 bovine poly-Q encoding genes (Q ≥ 5) and within this group, 26 genes with orthologs in both human and mouse that did not contain poly-Q repeats. The bovine poly-Q encoding genes typically had ubiquitous expression patterns although there was bias towards expression in epithelia, brain and testes. They were also characterised by unusually large sizes. Analysis of gene ontology terms revealed that the encoded proteins were strongly enriched for functions associated with transcriptional regulation and many contributed to physical interaction networks in the nucleus where they presumably act cooperatively in transcriptional regulatory complexes. In addition, the coding sequence CAG repeats in some bovine genes impacted mRNA splicing thereby generating unusual transcriptional diversity, which in at least one instance was tissue-specific. The poly-Q encoding genes were prioritised using multiple criteria for their likelihood of being polymorphic and then the highest ranking group was experimentally tested for polymorphic variation within a cattle diversity panel. Extensive and meiotically stable variation was identified

  3. Regulatory Interactions of Csr Components: the RNA Binding Protein CsrA Activates csrB Transcription in Escherichia coli

    OpenAIRE

    Gudapaty, Seshagirirao; Suzuki, Kazushi; Wang, Xin; Babitzke, Paul; Romeo, Tony

    2001-01-01

    The global regulator CsrA (carbon storage regulator) of Escherichia coli is a small RNA binding protein that represses various metabolic pathways and processes that are induced in the stationary phase of growth, while it activates certain exponential phase functions. Both repression and activation by CsrA involve posttranscriptional mechanisms, in which CsrA binding to mRNA leads to decreased or increased transcript stability, respectively. CsrA also binds to a small untranslated RNA, CsrB, f...

  4. Surfactant Protein-A as an Anti-Inflammatory Component in the Amnion: Implications for Human Pregnancy1

    OpenAIRE

    Lee, Deug-Chan; Romero, Roberto; Kim, Chong Jai; Chaiworapongsa, Tinnakorn; Tarca, Adi L.; Lee, JoonHo; Suh, Yeon-Lim; Mazaki-Tovi, Shali; Vaisbuch, Edi; Mittal, Pooja; Draghici, Sorin; Erez, Offer; Kusanovic, Juan Pedro; Hassan, Sonia S; Kim, Jung-Sun

    2010-01-01

    The mechanism of mouse parturition is thought to involve myometrial infiltration by amniotic fluid (AF) macrophages, activated by surfactant protein-A (SP-A). In humans, the concentration of AF SP-A decreases during labor, and no fetal macrophages are found in the myometrium after labor. Therefore, it appears that the mechanisms of labor in mice and humans are different. We investigated a potential role for SP-A in human pregnancy and parturition by examining SP-A expression patterns in AF an...

  5. The TatA component of the twin-arginine protein transport system forms channel complexes of variable diameter

    OpenAIRE

    Gohlke, Ulrich; Pullan, Lee; McDevitt, Christopher A.; Porcelli, Ida; de Leeuw, Erik; Palmer, Tracy; Saibil, Helen R.; Berks, Ben C.

    2005-01-01

    The Tat system mediates Sec-independent transport of folded precursor proteins across the bacterial plasma membrane or the chloroplast thylakoid membrane. Tat transport involves distinct high-molecular-weight TatA and TatBC complexes. Here we report the 3D architecture of the TatA complex from Escherichia coli obtained by single-particle electron microscopy and random conical tilt reconstruction. TatA forms ring-shaped structures of variable diameter in which the internal channels are large e...

  6. A Two-Component DNA-Prime/Protein-Boost Vaccination Strategy for Eliciting Long-Term, Protective T Cell Immunity against Trypanosoma cruzi

    OpenAIRE

    Shivali Gupta; Garg, Nisha J.

    2015-01-01

    In this study, we evaluated the long-term efficacy of a two-component subunit vaccine against Trypanosoma cruzi infection. C57BL/6 mice were immunized with TcG2/TcG4 vaccine delivered by a DNA-prime/Protein-boost (D/P) approach and challenged with T. cruzi at 120 or 180 days post-vaccination (dpv). We examined whether vaccine-primed T cell immunity was capable of rapid expansion and intercepting the infecting T. cruzi. Our data showed that D/P vaccine elicited CD4+ (30-38%) and CD8+ (22-42%) ...

  7. Search for novel targets of the PII signal transduction protein in Bacteria identifies the BCCP component of acetyl-CoA carboxylase as a PII binding partner.

    Science.gov (United States)

    Rodrigues, Thiago E; Gerhardt, Edileusa C M; Oliveira, Marco A; Chubatsu, Leda S; Pedrosa, Fabio O; Souza, Emanuel M; Souza, Gustavo A; Müller-Santos, Marcelo; Huergo, Luciano F

    2014-02-01

    The PII family comprises a group of widely distributed signal transduction proteins. The archetypal function of PII is to regulate nitrogen metabolism in bacteria. As PII can sense a range of metabolic signals, it has been suggested that the number of metabolic pathways regulated by PII may be much greater than described in the literature. In order to provide experimental evidence for this hypothesis a PII protein affinity column was used to identify PII targets in Azospirillum brasilense. One of the PII partners identified was the biotin carboxyl carrier protein (BCCP), a component of the acetyl-CoA carboxylase which catalyses the committed step in fatty acid biosynthesis. As BCCP had been previously identified as a PII target in Arabidopsis thaliana we hypothesized that the PII -BCCP interaction would be conserved throughout Bacteria. In vitro experiments using purified proteins confirmed that the PII -BCCP interaction is conserved in Escherichia coli. The BCCP-PII interaction required MgATP and was dissociated by increasing 2-oxoglutarate. The interaction was modestly affected by the post-translational uridylylation status of PII ; however, it was completely dependent on the post-translational biotinylation of BCCP. PMID:24329683

  8. Enhancer-binding proteins with a forkhead-associated domain and the sigma(54) regulon in Myxococcus xanthus fruiting body development

    DEFF Research Database (Denmark)

    Jelsbak, Lars; Givskov, Michael Christian; Kaiser, D.

    2005-01-01

    In response to starvation, Myxococcus xanthus initiates a developmental program that results in the formation of spore-filled, multicellular fruiting bodies. Many developmentally regulated genes in M. xanthus are transcribed from sigma(54) promoters, and these genes require enhancer-binding prote......In response to starvation, Myxococcus xanthus initiates a developmental program that results in the formation of spore-filled, multicellular fruiting bodies. Many developmentally regulated genes in M. xanthus are transcribed from sigma(54) promoters, and these genes require enhancer...... these genes, Mx4885, caused a cell autonomous aggregation and sporulation defect. In-frame deletion mutants showed that the FHA domain is necessary for proper Mx4885 function. The altered pattern of developmental gene expression in the mutant implied that Mx4885 is on the pathway of response to the...

  9. THE EFFECTS OF PRE-AND POST-EXERCISE WHEY VS. CASEIN PROTEIN CONSUMPTION ON BODY COMPOSITION AND PERFORMANCE MEASURES IN COLLEGIATE FEMALE ATHLETES

    Directory of Open Access Journals (Sweden)

    olin D. Wilborn

    2013-03-01

    Full Text Available Two of the most popular forms of protein on the market are whey and casein. Both proteins are derived from milk but each protein differs in absorption rate and bioavailability, thus it is possible that each type of protein may contribute differently to the adaptations elicited through resistance training. Therefore, the purpose of this study was to investigate the potential effects of ingestion of two types of protein in conjunction with a controlled resistance training program in collegiate female basketball players. Sixteen NCAA Division III female basketball players were matched according to body mass and randomly assigned in a double-blind manner to consume 24 g whey protein (WP (N = 8, 20.0 ± 1.9 years, 1.58 ± 0.27 m, 66. 0 ± 4.9 kg, 27.0 ± 4.9 %BF or 24 g casein protein (CP (N = 8, 21.0 ± 2.8 years, 1.53 ± 0.29 m, 68.0 ± 2.9 kg, 25.0 ± 5.7 %BF immediately pre- and post-exercise for eight weeks. Subjects participated in a supervised 4-day per week undulating periodized training program. At 0 and 8 weeks, subjects underwent DXA body composition analysis, and at 0 and 8 weeks underwent one repetition maximum (1RM strength, muscle endurance, vertical jump, 5-10-5 agility run, and broad jump testing sessions. Data were analyzed using repeated measures ANOVA, and presented as mean ± SD changes from baseline after 60 days. No significant group x time interaction effects were observed among groups in changes in any variable (p > 0.05. A significant time effect was observed for body fat (WP: -2.0 ± 1.1 %BF; CP: -1.0 ± 1.6 %BF, p < 0.001, lean mass (WP: 1.5 ± 1.0 kg; CP: 1. 4 ± 1.0 kg, p < 0.001, fat mass (WP: -1.3 ± 1.2 kg; CP: -0.6 ± 1.4 kg, p < 0.001, leg press 1RM (WP: 88.7 ± 43.9 kg; CP: 90.0 ± 48.5 kg, p < 0.001, bench press 1RM (WP: 7.5 ± 4.6 kg; CP: 4.3 ± 4.5 kg, p = 0.01, vertical jump (WP: 4.1 ± 1.8 cm; CP: 3.5 ± 7.6 cm, p < 0.001, 5-10-5 (WP: -0.3 ± 0.2 sec; CP: -0.09 ± 0.42 sec, p < 0.001, and broad jump (WP: 10

  10. Developmental Programming: Effect of Prenatal Steroid Excess on Intraovarian Components of Insulin Signaling Pathway and Related Proteins in Sheep1

    Science.gov (United States)

    Ortega, Hugo H.; Rey, Florencia; Velazquez, Melisa M.L.; Padmanabhan, Vasantha

    2010-01-01

    Prenatal testosterone (T) excess increases ovarian follicular recruitment, follicular persistence, insulin resistance, and compensatory hyperinsulinemia. Considering the importance of insulin in ovarian physiology, in this study, using prenatal T- and dihydrotestosterone (DHT, a nonaromatizable androgen)-treated female sheep, we tested the hypothesis that prenatal androgen excess alters the intraovarian insulin signaling cascade and metabolic mediators that have an impact on insulin signaling. Changes in ovarian insulin receptor (INSRB), insulin receptor substrate 1 (IRS1), mammalian target of rapamycin (MTOR), phosphatidylinositol 3-kinase (PIK3), peroxisome proliferator-activated receptor-gamma (PPARG), and adiponectin proteins were determined at fetal (Days 90 and 140), postpubertal (10 mo), and adult (21 mo) ages by immunohistochemistry. Results indicated that these proteins were expressed in granulosa, theca, and stromal compartments, with INSRB, IRS1, PPARG, and adiponectin increasing in parallel with advanced follicular differentiation. Importantly, prenatal T excess induced age-specific changes in PPARG and adiponectin expression, with increased PPARG expression evident during fetal life and decreased antral follicular adiponectin expression during adult life. Comparison of developmental changes in prenatal T and DHT-treated females found that the effects on PPARG were programmed by androgenic actions of T, whereas the effects on adiponectin were likely by its estrogenic action. These results suggest a role for PPARG in the programming of ovarian disruptions by prenatal T excess, including a decrease in antral follicular adiponectin expression and a contributory role for adiponectin in follicular persistence and ovulatory failure. PMID:20147730

  11. [The influence of procaine electrophoresis and mesotherapy on the indices of body components and biochemical characteristics in the subjects above 30 years of age].

    Science.gov (United States)

    Turova, E A; Konchugova, T V; Balaban, E I; Fadeeva, N I; Golovach, A V; Teniaeva, E A

    2013-01-01

    This study has demonstrated the effectiveness of a 2% procaine solution given with a view to improving metabolic parameters of aging, such as the body weight and composition along with the lipid profile in the patients varying in age from 30 years to 75 years. The study involved 95 patients treated with a 2% procaine solution that was administered by different methods (electrophoresis and mesotherapy) into several points of the collar region and upper back. The control group was comprised of the patients who received placebo by means of galvanization and mesotherapy. The results of the study indicate that the introduction of a 2% procaine solution in the treatment of metabolic disorders effectively improves selected metabolic characteristics of ageing and thereby helps to reduce the biological age in comparison with the control patients treated with placebo. PMID:23718080

  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.

    Science.gov (United States)

    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 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

  13. Estimation of Genetic Variance Components Including Mutation and Epistasis using Bayesian Approach in a Selection Experiment on Body Weight in Mice

    DEFF Research Database (Denmark)

    Widyas, Nuzul; Jensen, Just; Nielsen, Vivi Hunnicke

    selected downwards and three lines were kept as controls. Bayesian statistical methods are used to estimate the genetic variance components. Mixed model analysis is modified including mutation effect following the methods by Wray (1990). DIC was used to compare the model. Models including mutation effect...... have better fit compared to the model with only additive effect. Mutation as direct effect contributes 3.18% of the total phenotypic variance. While in the model with interactions between additive and mutation, it contributes 1.43% as direct effect and 1.36% as interaction effect of the total variance...

  14. Effect of Vegetable Protein and Oil Supplementation on Growth Performance and Body Composition of Russian Sturgeon Juveniles (Acipenser gueldenstaedtii Brandt, 1833) at Low temperatures

    OpenAIRE

    , Erdal Şener; Mustafa Yıldız; (1), Esra Savaş

    2006-01-01

    Effect of different feed compositions on growth performance and body composition of Russian sturgeon (Acipenser gueldenstaedtii) juveniles was studied. One control and three experimental diets were prepared with different ratios of soybean meal (20%, 10.5%, 33%, and 27%) sunflower meal (7%, 8.5%, 14.5%, and 10.5%) and corn meal (10%, 20.5%, 15% and 7.5%) substitute fish meal (40%, 35%, 25%, and 30%) as a protein source. Corn oil (2.5% in Group I and 10% in Group II) and sunflower oil (2.5% in...

  15. Pathologically Collapsed Vertebral Body

    Directory of Open Access Journals (Sweden)

    Reza Saadat Mostafavi

    2010-05-01

    Full Text Available An 8-year-old boy, a case of CGD, presenting with quadriparesis "nFindings: Collapsed contiguous vertebral bodies"nSpared disks"nEpidural components extending one level above and below the involved vertebral bodies"nSignal of involved vertebral bodies: low on T1W and high on T2W image

  16. Change in proportional protein intake in a 10-week energy-restricted low- or high-fat diet, in relation to changes in body size and metabolic factors

    DEFF Research Database (Denmark)

    Stocks, Tanja; Taylor, Moira A; Ängquist, Lars;

    2013-01-01

    Objective: To investigate in a secondary analysis of a randomised trial the effects of a low-/high-fat diet and reported change from baseline in energy% from protein (prot%), in relation to changes in body size and metabolic factors. Methods: Obese adults (n = 771) were randomised to a 600 kcal...... increased the percentage energy intake from protein showed the greatest reduction in weight and cholesterol, and a triglyceride reduction equally large to that of participants on a high-fat diet. Copyright © 2013 S. Karger GmbH, Freiburg....... energy-deficient low-fat (20-25 fat%) or high-fat (40-45 fat%) diet over 10 weeks. Dietary intake data at baseline and during the intervention were available in 585 completers. We used linear regression to calculate the combined effects of randomised group and groups of prot% change (2) on outcomes...

  17. Pse-in-One: a web server for generating various modes of pseudo components of DNA, RNA, and protein sequences.

    Science.gov (United States)

    Liu, Bin; Liu, Fule; Wang, Xiaolong; Chen, Junjie; Fang, Longyun; Chou, Kuo-Chen

    2015-07-01

    With the avalanche of biological sequences generated in the post-genomic age, one of the most challenging problems in computational biology is how to effectively formulate the sequence of a biological sample (such as DNA, RNA or protein) with a discrete model or a vector that can effectively reflect its sequence pattern information or capture its key features concerned. Although several web servers and stand-alone tools were developed to address this problem, all these tools, however, can only handle one type of samples. Furthermore, the number of their built-in properties is limited, and hence it is often difficult for users to formulate the biological sequences according to their desired features or properties. In this article, with a much larger number of built-in properties, we are to propose a much more flexible web server called Pse-in-One (http://bioinformatics.hitsz.edu.cn/Pse-in-One/), which can, through its 28 different modes, generate nearly all the possible feature vectors for DNA, RNA and protein sequences. Particularly, it can also generate those feature vectors with the properties defined by users themselves. These feature vectors can be easily combined with machine-learning algorithms to develop computational predictors and analysis methods for various tasks in bioinformatics and system biology. It is anticipated that the Pse-in-One web server will become a very useful tool in computational proteomics, genomics, as well as biological sequence analysis. Moreover, to maximize users' convenience, its stand-alone version can also be downloaded from http://bioinformatics.hitsz.edu.cn/Pse-in-One/download/, and directly run on Windows, Linux, Unix and Mac OS. PMID:25958395

  18. Feasibility and efficacy of an isocaloric high-protein vs. standard diet on insulin requirement, body weight and metabolic parameters in patients with type 2 diabetes on insulin therapy

    DEFF Research Database (Denmark)

    Luger, M; Holstein, B; Schindler, K;

    2013-01-01

    To determine the feasibility and efficacy of a high-protein diet compared with a standard diet aiming for weight maintenance in insulin treated type-2 diabetic patients on insulin requirement, body weight and metabolic parameters over 12 weeks.......To determine the feasibility and efficacy of a high-protein diet compared with a standard diet aiming for weight maintenance in insulin treated type-2 diabetic patients on insulin requirement, body weight and metabolic parameters over 12 weeks....

  19. Effect of gamma irradiation on the total nitrogen and protein content in body during different stages of silkworm development

    International Nuclear Information System (INIS)

    The aim was to determine the effect of gamma irradiation of eggs of silk moth in B2 stage in doses of 1.00, 2.00 and 3.00 Gy on the changes of total nitrogen and protein content during different stages of Bombyx mori L. development. Highest levels of total nitrogen and protein were found in silk gland 14.032-14.355 mg%, followed by pupae - 7.448-8.092 and 46.550-48.906 mg%, moths after egg laying - 6.650-7.825 and 41.563-48.906 mg% and silkworm hemolymph - 6.920-6.980 and 43.250-43.625 mg%, respectively. The irradiation of eggs with 2.00 and 3,00 Gy gamma rays stimulated the increase of total nitrogen and protein content in silk gland by 6.66-7.3% compared to non-irradiated eggs of the same breed. 14 refs., 3 tabs. (author)

  20. The mouse mismatch repair protein, MSH3, is a nucleoplasmic protein that aggregates into denser nuclear bodies under conditions of stress.

    NARCIS (Netherlands)

    Holt, I.; Thanh Lam, L.; Tome, S.; Wansink, D.G.; Riele, H. te; Gourdon, G.; Morris, G.E.

    2011-01-01

    The mismatch repair protein, MSH3, together with MSH2, forms the MutSbeta heterodimer which recognizes and repairs base pair mismatches and larger insertion/deletion loops in DNA. Lack of specific antibodies against mouse MSH3 has hampered studies of its expression and localization. Mouse MSH3 is no