Sample records for antifreeze

  1. Antifreeze poisoning (United States)

    The poisonous ingredients in antifreeze are: Ethylene glycol Methanol Propylene glycol ... For ethylene glycol: Death may occur within the first 24 hours. If ... little as 2 tablespoons (1 ounce or 30 milliliters) can kill a ...

  2. Synthetic antifreeze peptide



    A synthetic antifreeze peptide and a synthetic gene coding for the antifreeze peptide have been produced. The antifreeze peptide has a greater number of repeating amino acid sequences than is present in the native antifreeze peptides from winter flounder upon which the synthetic antifreeze peptide was modeled. Each repeating amino acid sequence has two polar amino acid residues which are spaced a controlled distance apart so that the antifreeze peptide may inhibit ice formation. The synthetic...

  3. Saccharide antifreeze compositions

    Energy Technology Data Exchange (ETDEWEB)

    Walters, Kent; Duman, John G; Serianni, Anthony S


    The invention provides an antifreeze glycolipid compounds and composition comprising a polysaccharide moiety of Formula I; ##STR00001## wherein D-Manp represents a D-mannopyranose moiety, D-Xylp represents a D-xylopyranose moiety, and n is about 5 to about 70; and one or more lipid moieties covalently linked to the polysaccharide moiety of Formula I or electrostatically associated with the polysaccaride moiety for Formula I. The antifreeze glycolipid compounds and compositions can be used for a variety of industrial, agricultural, medical, and cosmetic applications where recrystallization-inhibition, cyroprotection, or cryopreservation is desired. The antifreeze glycolipid compounds or compositions can be used as, for example, as cryoprotectants for tissue preservation and transplantation, improving the texture of processed frozen food and frozen meats, frostbit protection, crop protection, and green alternatives for land vehicle antifreeze and aircraft de-icing.

  4. Antifreeze Proteins of Bacteria

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 12; Issue 12. Antifreeze Proteins of Bacteria. M K Chattopadhyay. General Article Volume 12 Issue 12 December 2007 pp 25-30. Fulltext. Click here to view fulltext PDF. Permanent link: ...

  5. Antifreeze Proteins of Bacteria

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 12; Issue 12. Antifreeze Proteins of Bacteria. M K Chattopadhyay. General Article Volume 12 Issue 12 December 2007 pp 25-30. Fulltext. Click here to view fulltext PDF. Permanent link: Keywords.

  6. Synthetic antifreeze peptide and synthetic gene coding for its production



    A synthetic antifreeze peptide and a synthetic gene coding for the antifreeze peptide have been produced. The antifreeze peptide has a greater number of repeating amino acid sequences than is present in the native antifreeze peptides from winter flounder upon which the synthetic antifreeze peptide was modeled. Each repeating amino acid sequence has two polar amino acid residues which are spaced a controlled distance apart so that the antifreeze peptide may inhibit ice formation. The synthetic...

  7. Antifreeze life cycle assessment (LCA

    Directory of Open Access Journals (Sweden)

    Kesić Jelena


    Full Text Available Antifreeze based on ethylene glycol is a commonly used commercial product The classification of ethylene glycol as a toxic material increased the disposal costs for used antifreeze and life cycle assessment became a necessity. Life Cycle Assessment (LCA considers the identification and quantification of raw materials and energy inputs and waste outputs during the whole life cycle of the analyzed product. The objectives of LCA are the evaluation of impacts on the environment and improvements of processes in order to reduce and/or eliminate waste. LCA is conducted through a mathematical model derived from mass and energy balances of all the processes included in the life cycle. In all energy processes the part of energy that can be transformed into some other kind of energy is called exergy. The concept of exergy considers the quality of different types of energy and the quality of different materials. It is also a connection between energy and mass transformations. The whole life cycle can be described by the value of the total loss of exergy. The physical meaning of this value is the loss of material and energy that can be used. The results of LCA are very useful for the analyzed products and processes and for the determined conditions under which the analysis was conducted. The results of this study indicate that recycling is the most satisfactory solution for the treatment of used antifreeze regarding material and energy consumption but the re-use of antifreeze should not be neglected as a solution.

  8. Antifreeze proteins of teleost fishes. (United States)

    Fletcher, G L; Hew, C L; Davies, P L


    Marine teleosts at high latitudes can encounter ice-laden seawater that is approximately 1 degrees C colder than the colligative freezing point of their body fluids. They avoid freezing by producing small antifreeze proteins (AFPs) that adsorb to ice and halt its growth, thereby producing an additional non-colligative lowering of the freezing point. AFPs are typically secreted by the liver into the blood. Recently, however, it has become clear that AFP isoforms are produced in the epidermis (skin, scales, fin, and gills) and may serve as a first line of defense against ice propagation into the fish. The basis for the adsorption of AFPs to ice is something of a mystery and is complicated by the extreme structural diversity of the five antifreeze types. Despite the recent acquisition of several AFP three-dimensional structures and the definition of their ice-binding sites by mutagenesis, no common ice-binding motif or even theme is apparent except that surface-surface complementarity is important for binding. The remarkable diversity of antifreeze types and their seemingly haphazard phylogenetic distribution suggest that these proteins might have evolved recently in response to sea level glaciation occurring just 1-2 million years ago in the northern hemisphere and 10-30 million years ago around Antarctica. Not surprisingly, the expression of AFP genes from different origins can also be quite dissimilar. The most intensively studied system is that of the winter flounder, which has a built-in annual cycle of antifreeze expression controlled by growth hormone (GH) release from the pituitary in tune with seasonal cues. The signal transduction pathway, transcription factors, and promoter elements involved in this process are just beginning to be characterized.

  9. Natural and Artificial Antifreeze Proteins (United States)

    Arai, Soichi; Hirao, Noriko

    In the blood of winter polar fish an antifreeze glycoprotein (AFGP) occurs which acts to protect the fish from freezing to death. The AFGP has a unique hydrophilic hydrophobic conformation, involved in non-colligative depression of the freezing temperature of water without altering the melting point of ice. This phenomenon is reportedly a reflection of the ice crystal growth inhibition by the adsorption of the AFGP onto a-axial surfaces of the ice crystal. The authors, on the other hand, have developed an enzymatically modified protein (EMG-12) by covalent attachment of leucine dodecyl ester to the C-terminal position of gelation with the aid of a reverse reaction catalyzed by a protease. EMG-12, having a hydrophilic-hydrophobic structure, is highly surface-active and acts to stabilize a supercooling state of water by antinucleation. Discussions are made on similarities and dissimilarities of structure-function relationships of these natural and artificial antifreeze proteins. The significance of using them as antifreeze agents is also discussed.

  10. A kinetic description of antifreeze glycoprotein activity. (United States)

    Burcham, T S; Osuga, D T; Yeh, Y; Feeney, R E


    The antifreeze glycoproteins (AFGP) of polar fish have the ability to depress the freezing temperature of water approximately 500 times the amount expected based on the number of AFGP molecules in solution; yet AFGP solutions have a purely colligative melting point depression. The difference of solution melting and freezing temperatures is the antifreeze activity of AFGP. One characteristic of AFGP activity that requires further examination is the effect of concentration on antifreeze activity, especially whether the activity saturates at high concentrations or the measured activity increases ad infinitum. This study first surveys the activity of the various antifreeze components from both Pagothenia borchgrevinki and the Arg-containing antifreeze glycoprotein from Eleginus gracilis (EgAF). It was found that all AFGP components examined have a plateau in activity at high concentration, but the actual value of the plateau activity differs between the different length AFGP components and between AFGP and EgAF. While the low molecular weight components of both AFGP and EgAF lose activity at deep supercooling, at high concentration activity is restored. The activity data is then shown to fit a reversible kinetic model of AFGP activity, and the coefficients obtained are used to compare the activity differences between AFGP components and between AFGP and EgAF. The model is also shown to describe the activity of the antifreeze protein of the fish Pseudopleuronectes americanus and the thermal hysteresis protein of the insect, Tenebrio molitor.

  11. Efficiency of Composite Binders with Antifreezing Agents (United States)

    Ogurtsova, Y. N.; Zhernovsky, I. V.; Botsman, L. N.


    One of the non-heating methods of cold-weather concreting is using concretes hardening at negative temperatures. This method consists in using chemical additives which reduce the freezing temperature of the liquid phase and provide for concrete hardening at negative temperatures. The non-heating cold-weather concreting, due to antifreezing agents, allows saving heat and electric energy at the more flexible work performance technology. At selecting the antifreezing components, the possibility of concreting at temperatures up to minus 20 °C and combination with a plasticizer contained in the composite binder were taken into account. The optimal proportions of antifreezing and complex agents produced by MC-Bauchemie Russia for fine-grained concretes were determined. So, the introduction of antifreezing and complex agents allows obtaining a structure of composite characteristic for cement stone in the conditions of below zero temperatures at using different binders; the hydration of such composite proceeded naturally. Low-water-demand binders (LWDB) based composites are characterized by a higher density and homogeneity due to a high dispersity of a binder and its complicated surface providing for a lot of crystallization centers. LWDB contains small pores keeping water in a liquid form and promoting a more complete hydration process.

  12. Antivirulence Properties of an Antifreeze Protein

    Directory of Open Access Journals (Sweden)

    Martin Heisig


    Full Text Available As microbial drug-resistance increases, there is a critical need for new classes of compounds to combat infectious diseases. The Ixodes scapularis tick antifreeze glycoprotein, IAFGP, functions as an antivirulence agent against diverse bacteria, including methicillin-resistant Staphylococcus aureus. Recombinant IAFGP and a peptide, P1, derived from this protein bind to microbes and alter biofilm formation. Transgenic iafgp-expressing flies and mice challenged with bacteria, as well as wild-type animals administered P1, were resistant to infection, septic shock, or biofilm development on implanted catheter tubing. These data show that an antifreeze protein facilitates host control of bacterial infections and suggest therapeutic strategies for countering pathogens.

  13. Antifreeze glycoprotein agents: structural requirements for activity. (United States)

    Carvajal-Rondanelli, Patricio A; Marshall, Sergio H; Guzman, Fanny


    Antifreeze glycoproteins (AFGPs) are considered to be the most efficient means to reduce ice damage to cell tissues since they are able to inhibit growth and crystallization of ice. The key element of antifreeze proteins is to act in a non-colligative manner which allows them to function at concentrations 300-500 times lowers than other dissolved solutes. During the past decade, AFGPs have demonstrated tremendous potential for many pharmaceutical and food applications. Presently, the only route to obtain AFGPs involves the time consuming and expensive process of isolation and purification from deep-sea polar fishes. Unfortunately, it is not amenable to mass production and commercial applications. The lack of understanding of the mechanism through which the AFGPs inhibit ice growth has also hampered the realization of industrial and biotechnological applications. Here we report the structural motifs that are essential for antifreeze activity of AFGPs, and propose a unified mechanism based on both recent studies of short alanine peptides and structure activity relationship of synthesized AFGPs. Copyright © 2011 Society of Chemical Industry.

  14. Antifreeze activity enhancement by site directed mutagenesis on an antifreeze protein from the beetle Rhagium mordax

    DEFF Research Database (Denmark)

    Friis, Dennis Steven; Kristiansen, Erlend; von Solms, Nicolas


    The ice binding motifs of insect antifreeze proteins (AFPs) mainly consist of repetitive TxT motifs aligned on a flat face of the protein. However, these motifs often contain non-threonines that disrupt the TxT pattern. We substituted two such disruptive amino acids located in the ice binding face...... of an AFP from Rhagium mordax with threonine. Furthermore, a mutant with an extra ice facing TxT motif was constructed. These mutants showed enhanced antifreeze activity compared to the wild type at low concentrations. However, extrapolating the data indicates that the wild type will become the most active...

  15. Antifreeze proteins enable plants to survive in freezing conditions

    Indian Academy of Sciences (India)

    Overwintering plants secrete antifreeze proteins (AFPs) to provide freezing tolerance. These proteins bind to and inhibit the growth of ice crystals that are formed in the apoplast during subzero temperatures. Antifreeze activity has been detected in more than 60 plants and AFPs have been purified from 15 of these, including ...

  16. Structural basis of antifreeze activity of a bacterial multi-domain antifreeze protein.

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    Chen Wang

    Full Text Available Antifreeze proteins (AFPs enhance the survival of organisms inhabiting cold environments by affecting the formation and/or structure of ice. We report the crystal structure of the first multi-domain AFP that has been characterized. The two ice binding domains are structurally similar. Each consists of an irregular β-helix with a triangular cross-section and a long α-helix that runs parallel on one side of the β-helix. Both domains are stabilized by hydrophobic interactions. A flat plane on the same face of each domain's β-helix was identified as the ice binding site. Mutating any of the smaller residues on the ice binding site to bulkier ones decreased the antifreeze activity. The bulky side chain of Leu174 in domain A sterically hinders the binding of water molecules to the protein backbone, partially explaining why antifreeze activity by domain A is inferior to that of domain B. Our data provide a molecular basis for understanding differences in antifreeze activity between the two domains of this protein and general insight on how structural differences in the ice-binding sites affect the activity of AFPs.

  17. Synthesis of Structures Related to Antifreeze Glycoproteins


    Fyrner, Timmy


    In this thesis, synthesis of structures related to antifreeze glycoproteins (AFGPs) are presented. Synthetic routes to a protected carbohydrate derivative, 2,3,4,6-tetra-O-benzyl-β-galactopyranosyl-(1→3)-2-deoxy-2-azido-4,6-di-O-benzyl-β-D-thio-1-galactopyranoside, and a tBu-Ala-Thr-Ala-Fmoc tripeptide, are described. These compounds are meant to be used in the assembly of AFGPs and analogues thereof. A Gal-GlcN disaccharide was synthesized via glycosylation between the donor, bromo-2-O-benzo...

  18. Antifreeze Proteins from Diverse Organisms and their Applications: An Overview. (United States)

    Cheung, Randy Chi Fai; Ng, Tzi Bun; Wong, Jack Ho


    Antifreeze proteins are ice-binding or ice-structuring proteins that prevent water from freezing by adsorbing to the ice surface and stopping the growth of minute ice crystals to large crystals in a non-colligative manner. The antifreeze proteins are found in species like fish, arthropods, plants, algae, fungi, yeasts and bacteria. The diversity, distribution and classification of antifreeze proteins were highlighted in this review. Antifreeze proteins help the organisms adapt to and survive in subzero temperature environments. The distribution of antifreeze proteins in different species appears to be the outcome of a combination of independent evolutionary events, probably the convergent evolution or horizontal gene transfer. Benefits can be derived from the frost resistance of these organisms. Their potential applications have been recognized in food processing, cryopreservation, cryosurgery, fishery and agricultural industries and anti-icing materials development. This review includes information on the current understanding of antifreeze proteins. A discussion on interactions and mechanisms involving ice recognition and adsorption was also included. Copyright© Bentham Science Publishers; For any queries, please email at

  19. Antifreeze Protein Mimetic Metallohelices with Potent Ice Recrystallization Inhibition Activity. (United States)

    Mitchell, Daniel E; Clarkson, Guy; Fox, David J; Vipond, Rebecca A; Scott, Peter; Gibson, Matthew I


    Antifreeze proteins are produced by extremophile species to control ice formation and growth, and they have potential applications in many fields. There are few examples of synthetic materials which can reproduce their potent ice recrystallization inhibition property. We report that self-assembled enantiomerically pure, amphipathic metallohelicies inhibited ice growth at just 20 μM. Structure-property relationships and calculations support the hypothesis that amphipathicity is the key motif for activity. This opens up a new field of metallo-organic antifreeze protein mimetics and provides insight into the origins of ice-growth inhibition.

  20. Renal conservation of antifreeze peptide in Antarctic eelpout, Rhigophila dearborni. (United States)

    Eastman, J T; De Vries, A L; Coalson, R E; Nordquist, R E; Boyd, R B


    In Antarctic notothenioid fishes large amounts (3% w/v) of small molecular weights of 2,600-23,500 and would be expected to be filtered into the urine, they remain in the blood because the kidneys of these fishes contain only aglomerular nephrons. Unlike the situation in most fishes, urine formation is the result of secretion rather than filtration and reabsorption. On the other hand, the peptide antifreezes in Northern Hemisphere fishes such as the winter flounder. Pseudopleuronectes americanus, are retained by the glomerular kidney even though inulin, of comparable weight, is rapidly filtered from the blood into the urine. The Antarctic eelpout (zoarcid), Rhigophila dearborni, which is unrelated to either the Antarctic notothenioids or P. americanus, also uses a peptide antifreeze (molecular weight 6,000) which is maintained at a concentration of 3% (w/v) in the blood plasma. We report here that the lack of antifreeze in the urine of R. dearborni probably reflects the fact that the glomeruli are not functional and cannot filter. We support this conclusion with morphological and physiological evidence and relate our findings to the conservation of biological antifreeze necessary for life in ice-laden polar waters.

  1. Antifreeze proteins enable plants to survive in freezing conditions

    Indian Academy of Sciences (India)


    Oct 20, 2014 ... 0.52°C. TH. Zhang et al. 2011. 4. Daucus carrota. Tap. Root extract. 36. Apoplastic. Polygalacturonase inhibitor protein. N-glycosylated,. TH-0.36 at. 150 μ ...... na R 2011 A framework for classification of antifreeze proteins in over wintering plants based on their sequence and structural features. J. Bioinform.

  2. Studies on new antifreeze protein from the psychrophilic diatom ...

    African Journals Online (AJOL)

    Antifreeze proteins (AFPs) are found in a wide range of species including fishes, plants, etc. They have very characteristic feature that inhibit the growth and recrystallization of ice that forms in intercellular spaces. Two expressed sequence tags (ESTs) were previously identified from salt stress cDNA library in Fragilariopsis ...

  3. In silico characterization of antifreeze proteins using computational ...

    Indian Academy of Sciences (India)

    In this paper, seventeen different fish Antifreeze Proteins (AFPs) retrieved from Swiss-Prot database are analysed and characterized using In silico tools. ... Department of Chemistry, Sri Chandrasekharendra Saraswathi Viswa Maha Vidyalaya (Deemed University), Enathur, Kanchipuram 631 561; EXCEL and Polymer ...


    Directory of Open Access Journals (Sweden)

    S. V. Gushchin


    Full Text Available Usage of chemical additives while executing concrete works at negative temperatures is considered as a convenient and economical method. Range of the used antifreeze additives is rather wide. A great number of new additives are advertised but their characteristics have not been practically studied. Evaluation of the antifreeze additive efficiency is unfortunately rather long process and it does not provide comprehensive data on concrete structure formation processes. Due to this development of rapid and comprehensive methodology for construction companies is urgently required.Freezing processes of antifreeze additive aqueous solutions and hardening of cement paste with them have been investigated in the paper. The paper proposes a methodology for determination of freezing point for aqueous solutions of chemical additives of various applications. Identity of  freezing point for a chemical additive aqueous solution and cement paste with an equal concentration of the additive in the paste pore fluid has been determined while taking  calcium nitrate and sodium formate additives as an example. The paper demonstrates the possibility to evaluate efficiency of antifreeze additive action on the basis of kinetics in temperature changes of the cement paste with additives by its consecutive freezing and defrosting.  A methodology for operational evaluation in the field of chemical additive application for concreting items at negative temperatures has been offered in the paper.  The methodology does not require  deficient and expensive test-equipment. It can be applied at ordinary construction companies and it is comprehensible for personnel of low-qualification.  The paper shows the possibility to develop an original methodology for designing concrete structure which is based on operating efficiency determinations  for single and integrated antifreeze additives.

  5. Potential Antifreeze Compounds in Present-Day Martian Seepage Groundwater

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    Jiin-Shuh Jean


    Full Text Available Is the recently found seepage groundwater on Mars pure H2O, or mixed with salts and other antifreeze compounds? Given the surface conditions of Mars, it is unlikely that pure water could either exist in its liquid state or have shaped Mars¡¦ fluid erosional landforms (gullies, channels, and valley networks. More likely is that Mars¡¦ seepage groundwater contains antifreeze and salt compounds that resist freezing and suppress evaporation. This model better accounts for Mars¡¦ enigmatic surface erosion. This paper suggests 17 antifreeze compounds potentially present in Martian seepage groundwater. Given their liquid state and physical properties, triethylene glycol, diethylene glycol, ethylene glycol, and 1,3-propylene glycol are advanced as the most likely candidate compounds. This paper also explores how a mixing of glycol or glycerol with salts in the Martian seepage groundwater may have lowered water¡¦s freezing point and raised its boiling point, with consequences that created fluid gully and channel erosion. Ethylene glycol and related hydrocarbon compounds have been identified in Martian and other interstellar meteorites. We suggest that these compounds and their proportions to water be included for detection in future explorations.

  6. Structure and application of antifreeze proteins from Antarctic bacteria. (United States)

    Muñoz, Patricio A; Márquez, Sebastián L; González-Nilo, Fernando D; Márquez-Miranda, Valeria; Blamey, Jenny M


    Antifreeze proteins (AFPs) production is a survival strategy of psychrophiles in ice. These proteins have potential in frozen food industry avoiding the damage in the structure of animal or vegetal foods. Moreover, there is not much information regarding the interaction of Antarctic bacterial AFPs with ice, and new determinations are needed to understand the behaviour of these proteins at the water/ice interface. Different Antarctic places were screened for antifreeze activity and microorganisms were selected for the presence of thermal hysteresis in their crude extracts. Isolates GU1.7.1, GU3.1.1, and AFP5.1 showed higher thermal hysteresis and were characterized using a polyphasic approach. Studies using cucumber and zucchini samples showed cellular protection when samples were treated with partially purified AFPs or a commercial AFP as was determined using toluidine blue O and neutral red staining. Additionally, genome analysis of these isolates revealed the presence of genes that encode for putative AFPs. Deduced amino acids sequences from GU3.1.1 (gu3A and gu3B) and AFP5.1 (afp5A) showed high similarity to reported AFPs which crystal structures are solved, allowing then generating homology models. Modelled proteins showed a triangular prism form similar to β-helix AFPs with a linear distribution of threonine residues at one side of the prism that could correspond to the putative ice binding side. The statistically best models were used to build a protein-water system. Molecular dynamics simulations were then performed to compare the antifreezing behaviour of these AFPs at the ice/water interface. Docking and molecular dynamics simulations revealed that gu3B could have the most efficient antifreezing behavior, but gu3A could have a higher affinity for ice. AFPs from Antarctic microorganisms GU1.7.1, GU3.1.1 and AFP5.1 protect cellular structures of frozen food showing a potential for frozen food industry. Modeled proteins possess a β-helix structure, and

  7. Antifreeze Glycoproteins Alter the Molecular Scale Surface Morphology of Ice (United States)

    Zepeda, Salvador; Orme, Christine A.; Qiu, Roger; Yeh, Yin


    Trematomas borchgrevinki live in the harsh super-cooled waters of the Antarctic. Critical to their survival are antifreeze glycoproteins (AFGPs) that further suppress the freezing temperature of their blood serum in addition to the colligative action of salts found in the ocean. These proteins also modify ice crystal growth habits as well as inhibit recrystallization in polycrystalline ice. To date many other types of antifreeze proteins have been identified in cold weather insects, plants, and other fish, but the exact mechanism is not entirely understood. The mechanism is non-colligative since only a few mg/ml are required for ice crystal growth inhibition and a non-equilibrium melting/freezing point hysteresis is observed. Atomic force microscopy (AFM) can yield a wealth of surface information that can reveal molecular scale information of biomineralization processes. We use AFM to directly probe the surface of ice crystals grown from the vapor in the pure phase and in the presence of growth inhibitors/modifiers, AFGPs. Results show that the AFGPs heavily pin the surface of ice.

  8. Ice growth in supercooled solutions of antifreeze glycoprotein. (United States)

    Harrison, K; Hallett, J; Burcham, T S; Feeney, R E; Kerr, W L; Yeh, Y

    Inhibition of ice growth in supercooled solution by certain proteins is vital to the survival of many living organisms. Some fish, native to both subzero northern and southern waters, have special proteins or glycoproteins in their blood serum that inhibit ice formation. Whereas these proteins have only a very small effect on the melting temperature of ice, the temperature of these fish can fall to nearly 1 K below the melting point before ice crystals grow. This phenomenon is called freezing hysteresis, in contrast to the normal colligative effect of solutes that depresses the equilibrium temperature, around which small changes lead to crystal growth or melting depending on sign. Some insects also exhibit a serum freezing hysteresis. We report the effects of different degrees of supercooling on the habit and rates of growth of ice crystals from solutions of these antifreeze glycoproteins (AFGPs). We find that the crystallization rate is up to five times greater than that in pure water.

  9. Inhibition of Condensation Frosting by Arrays of Hygroscopic Antifreeze Drops. (United States)

    Sun, Xiaoda; Damle, Viraj G; Uppal, Aastha; Linder, Rubin; Chandrashekar, Sriram; Mohan, Ajay R; Rykaczewski, Konrad


    The formation of frost and ice can have negative impacts on travel and a variety of industrial processes and is typically addressed by dispensing antifreeze substances such as salts and glycols. Despite the popularity of this anti-icing approach, some of the intricate underlying physical mechanisms are just being unraveled. For example, recent studies have shown that in addition to suppressing ice formation within its own volume, an individual salt saturated water microdroplet forms a region of inhibited condensation and condensation frosting (RIC) in its surrounding area. This occurs because salt saturated water, like most antifreeze substances, is hygroscopic and has water vapor pressure at its surface lower than water saturation pressure at the substrate. Here, we demonstrate that for macroscopic drops of propylene glycol and salt saturated water, the absolute RIC size can remain essentially unchanged for several hours. Utilizing this observation, we demonstrate that frost formation can be completely inhibited in-between microscopic and macroscopic arrays of propylene glycol and salt saturated water drops with spacing (S) smaller than twice the radius of the RIC (δ). Furthermore, by characterizing condensation frosting dynamics around various hygroscopic drop arrays, we demonstrate that they can delay complete frosting over of the samples 1.6 to 10 times longer than films of the liquids with equivalent volume. The significant delay in onset of ice nucleation achieved by dispensing propylene glycol in drops rather than in films is likely due to uniform dilution of the drops driven by thermocapillary flow. This transport mode is absent in the films, leading to faster dilution, and with that facilitated homogeneous nucleation, near the liquid-air interface.

  10. Blocking rapid ice crystal growth through nonbasal plane adsorption of antifreeze proteins. (United States)

    Olijve, Luuk L C; Meister, Konrad; DeVries, Arthur L; Duman, John G; Guo, Shuaiqi; Bakker, Huib J; Voets, Ilja K


    Antifreeze proteins (AFPs) are a unique class of proteins that bind to growing ice crystal surfaces and arrest further ice growth. AFPs have gained a large interest for their use in antifreeze formulations for water-based materials, such as foods, waterborne paints, and organ transplants. Instead of commonly used colligative antifreezes such as salts and alcohols, the advantage of using AFPs as an additive is that they do not alter the physicochemical properties of the water-based material. Here, we report the first comprehensive evaluation of thermal hysteresis (TH) and ice recrystallization inhibition (IRI) activity of all major classes of AFPs using cryoscopy, sonocrystallization, and recrystallization assays. The results show that TH activities determined by cryoscopy and sonocrystallization differ markedly, and that TH and IRI activities are not correlated. The absence of a distinct correlation in antifreeze activity points to a mechanistic difference in ice growth inhibition by the different classes of AFPs: blocking fast ice growth requires rapid nonbasal plane adsorption, whereas basal plane adsorption is only relevant at long annealing times and at small undercooling. These findings clearly demonstrate that biomimetic analogs of antifreeze (glyco)proteins should be tailored to the specific requirements of the targeted application.

  11. [Antifreeze glycoproteins in fishes: structure, mode of action and possible applications]. (United States)

    Wöhrmann, A


    Two types of antifreezes have been isolated from polar and northern temperate fishes so far. They are either glycopeptides or peptides. Whereas these proteins have only a very small effect on the melting temperature of ice, the temperature of these fish can fall to nearly 1 degree below the melting point before ice crystals grow. This phenomenon is called thermal hysteresis, in contrast to the normal colligative effect of solutes. All Antarctic notothenioids (perches) investigated so far have the typical antifreeze glycoproteins (AFGP) with the tripeptide Ala-Ala-Thr and the disaccharide Gal-GalNAc. In the Antarctic silverfish Pleuragramma antarcticum there could be found a novel GlcNAc containing antifreeze glycoprotein, the PAGP. The antifreezes not only lower the freezing temperature, but they also retard recrystallization on frozen storage. Antifreeze proteins thus could be useful for biotechnology and cryomedicine in the future. Since some are now synthesized chemically or by genetic engineering, they no longer have to be isolated from fish blood.


    Directory of Open Access Journals (Sweden)

    Bhavana Sharma


    Full Text Available Seabuckthorn (Hippophae rhamnoides, a cold desert shrub, can survive freezing temperature conditions and is considered as frost and drought tolerant. Earlier, antifreeze activity was reported in seabuckthorn seedlings grown under laboratory conditions. No reports are available on the cold hardiness of this huge bioresource available naturally in the Himalayan region. Detection of antifreeze activity in leaves and berries (splat assay and nanoliter osmometer confirmed the presence of putative antifreeze proteins (AFPs which may help in the survival of this plant under freezing conditions. Flower shaped ice crystals were observed in both leaves and berries while hexagonal ice crystals in seedlings indicated comparatively higher antifreeze activity in the wild parts. Splat assay results confirmed highest IRI activity in leaf (a 2.75 fold decrease in mean grain size of ice crystal after annealing followed by berry (with 1.75 fold decrease and least in the seedlings (with 1.5 fold decrease. Gel filtration chromatography resolved leaf fractions exhibited antifreeze activity in 34, 36 and 41 kDa while in berry fractions a 41 kDa polypeptide showed antifreeze activity. This is the first report showing presence of AFPs in (H. rhamnoides leaf and berry. Shotgun proteomic analysis using Q-Exactive Orbitrap High Resolution Mass Spectrometer and functional annotation of leaf and berry proteins revealed their association with primary, secondary metabolism, defence/stress, redox regulation, signalling and structural remodelling majority of which are affected during cold stress. Further purification of these AFPs could open gates for commercial utilization of this plant growing abundantly in Himalayan regions of India.

  13. Cloning, expression, and activity of type IV antifreeze protein from cultured subtropical olive flounder (Paralichthys olivaceus

    Directory of Open Access Journals (Sweden)

    Jong Kyu Lee


    Full Text Available Abstract Antifreeze proteins (AFPs lower the freezing point but not the melting point of aqueous solutions by inhibiting the growth of ice crystals via an adsorption-inhibition mechanism. However, the function of type IV AFP (AFP IV is questionable, as its antifreeze activity is on the verge of detectable limits, its physiological concentration in adult fish blood is too low to function as a biological antifreeze, and its homologues are present even in fish from tropic oceans as well as freshwater. Therefore, we speculated that AFP IV may have gained antifreeze activity not by selective pressure but by chance. To test this hypothesis, we cloned, expressed, and assayed AFP IV from cultured subtropical olive flounder (Paralichthys olivaceus, which do not require antifreeze protein for survival. Among the identified expressed sequence tags of the flounder liver sample, a 5′-deleted complementary DNA (cDNA sequence similar to the afp4 gene of the longhorn sculpin was identified, and its full-length cDNA and genome structure were examined. The deduced amino acid sequence of flounder AFP IV shared 55, 53, 52, and 49 % identity with those of Pleuragramma antarcticum, Myoxocephalus octodecemspinosus, Myoxocephalus scorpius, and Notothenia coriiceps, respectively. Furthermore, the genomic structure of this gene was conserved with those of other known AFP IVs. Notably, the recombinant AFP IV showed a weak but distinct thermal hysteresis of 0.07 ± 0.01 °C at the concentration of 0.5 mg/mL, and ice crystals in an AFP IV solution grew star-shaped, which are very similar to those obtained from other polar AFP IVs. Taken together, our results do not support the hypothesis of evolution of AFP IV by selective pressure, suggesting that the antifreeze activity of AFP IV may have been gained by chance.

  14. Two domains of RD3 antifreeze protein diffuse independently. (United States)

    Holland, Nolan B; Nishimiya, Yoshiyuki; Tsuda, Sakae; Sönnichsen, Frank D


    Antifreeze proteins (AFPs) make up a class of structurally diverse proteins that help to protect many organisms from freezing temperatures by inhibiting ice crystal growth at temperatures below the colligative freezing point. AFPs are typically small proteins with a relatively flat, slightly hydrophobic binding region that matches the lattice structure of a specific ice crystal plane. The only known two-domain AFP is RD3 from the Antarctic eel pout. It consists of two nearly identical type III domains connected by a nine-residue linker. This protein exhibits higher activity than the single-domain protein at low concentrations. The initial solution structure of RD3 revealed that the domains were aligned so that the binding regions were nearly coplanar, effectively doubling the surface area for binding. A more recent report suggests that the domains may not be aligned in solution but rather diffuse independently. To resolve the issue, we have measured the NMR residual dipolar couplings using alignment media of stretched gels and filamentous phage to determine the relative orientation of the domains. We find that the two domains of RD3 are free to move relative to each other, within the constraint of the flexible nine-residue linker. Our data show that there is no strongly preferred alignment in solution. Furthermore, the flexibility and length of the linker are sufficient to allow the two domains to have their binding faces in the same orientation and coplanar for simultaneous binding to an ice crystal surface.

  15. Solvation structure of ice-binding antifreeze proteins (United States)

    Hansen-Goos, Hendrik; Wettlaufer, John


    Antifreeze proteins (AFPs) can be found in organisms which survive at subzero temperatures. They were first discovered in polar fishes since the 1950's [1] and have been isolated meanwhile also from insects, plants, and bacteria. While AFPs shift the freezing point of water below the bulk melting point and hence can prevent recrystallization; the effect is non-colligative and there is a pronounced hysteresis between freezing and melting. For many AFPs it is generally accepted that they function through an irreversible binding to the ice-water interface which leads to a piecewise convex growth front with a lower nonequilibrium freezing point due to the Kelvin effect. Recent molecular dynamics simulations of the AFP from Choristoneura fumiferana reveal that the solvation structures of water at ice-binding and non-ice-binding faces of the protein are crucial for understanding how the AFP binds to the ice surface and how it is protected from being overgrown [2]. We use density functional theory of classical fluids in order to assess the microscopic solvent structure in the vicinity of protein faces with different surface properties. With our method, binding energies of different protein faces to the water-ice-interface can be computed efficiently in a simplified model. [1] Y. Yeh and R.E. Feeney, Chem. Rev. 96, 601 (1996). [2] D.R. Nutt and J.C. Smith, J. Am. Chem. Soc. 130, 13066 (2008).

  16. Anchored Clathrate Waters Bind Antifreeze Proteins to Ice

    Energy Technology Data Exchange (ETDEWEB)

    C Garnham; R Campbell; P Davies


    The mechanism by which antifreeze proteins (AFPs) irreversibly bind to ice has not yet been resolved. The ice-binding site of an AFP is relatively hydrophobic, but also contains many potential hydrogen bond donors/acceptors. The extent to which hydrogen bonding and the hydrophobic effect contribute to ice binding has been debated for over 30 years. Here we have elucidated the ice-binding mechanism through solving the first crystal structure of an Antarctic bacterial AFP. This 34-kDa domain, the largest AFP structure determined to date, folds as a Ca{sup 2+}-bound parallel beta-helix with an extensive array of ice-like surface waters that are anchored via hydrogen bonds directly to the polypeptide backbone and adjacent side chains. These bound waters make an excellent three-dimensional match to both the primary prism and basal planes of ice and in effect provide an extensive X-ray crystallographic picture of the AFP{vert_ellipsis}ice interaction. This unobstructed view, free from crystal-packing artefacts, shows the contributions of both the hydrophobic effect and hydrogen bonding during AFP adsorption to ice. We term this mode of binding the 'anchored clathrate' mechanism of AFP action.

  17. Antifreeze polymeric additives for fuels; Aditivos polimericos anticongelantes para combustiveis

    Energy Technology Data Exchange (ETDEWEB)

    Muniz, Aline S.; Carvalho, Agne Roani de; Sakae, George Hideki; Oliveira, Angelo R.S.; Cesar-Oliveira, Maria Aparecida F. [Universidade Federal do Parana - UFPR - Departamento de Quimica - LABPOL-Laboratorio de Polimeros Sinteticos, Centro Politecnico, Curitiba, PR (Brazil)], e-mails:,


    Owing to the current interest in the reduction of environmental pollution, several researchers are seeking renewable sources of energy which can at least partially replace combustibles derived from petroleum. Diesel oil is the combustible that most seriously pollutes the environment and is thus the biodiesel that is being considered as a fuel which can be replaced by a renewable combustible; this can possibly be used in diesel engines without any modifications. However, certain problems have to be overcome with regard to the temperature at which the biodiesel should be stored and used, since there is a tendency for biodiesel to solidify at low temperatures. This suggests that there is a need for the use of anti-freeze additives. This work behind the main focus additives with only 25 ppm, were able to reduce the pour point of fuel, achieving significant results, for example, the additive M14A18 lowered the pour point (PP) of B20 to -20 degree C, showing that the use of increasing amounts of biodiesel to diesel can aggregate. The main focus of work behind the development of additives that with only 25 ppm, were able to reduce the pour point of fuel, producing significant results such as those obtained with the use of additive M14A18 which lowered the pour point of the B20 to -20 degree C, showing the possibility of using increasing amounts of biodiesel added to diesel. (author)



    Wilson, P.; Devries, A.


    Glycopeptide and peptide antifreeze agents are present in the body fluids of polar fishes and allow them to avoid freezing in ice-laden seawater. These antifreezes lower the freezing point 200 times more than predicted by colligative relations, but have little effect on the melting point of ice. They bind to ice and appear to inhibit growth by increasing the curvature of growth steps on the ice crystal surface. Such a growth would result in a substantial increase in the roughness of the surfa...

  19. Structural characteristics of a novel antifreeze protein from the longhorn beetle Rhagium inquisitor

    DEFF Research Database (Denmark)

    Kristiansen, E; Ramløv, Hans; Højrup, Peter


    beetle Rhagium inquisitor is known to express AFPs in its body fluids. In this work we report on the primary structure and structural characteristics of a 12.8 kDa AFP from this beetle (RiAFP). It has a high capacity to evoke antifreeze activity as compared to other known insect AFPs......Antifreeze proteins (AFPs) are characterized by their capacity to inhibit the growth of ice and are produced by a variety of polar fish, terrestrial arthropods and other organisms inhabiting cold environments. This capacity reflects their role as stabilizers of supercooled body fluids. The longhorn...

  20. Antifreeze proteins: Adsorption to ice, silica and gas hydrates

    Energy Technology Data Exchange (ETDEWEB)

    Zeng, Huang; Brown, Alan; Wathen, Brent; Ripmeester, John A.; Walker, VIrginia K.


    Certain organisms survive under freezing conditions that could otherwise prove fatal by the synthesis of antifreeze proteins (AFPs). AFPs adsorb to the surface of microscopic ice crystals and prevent further ice growth, resulting in a noncolligative freezing point depression. Type I AFP from the winter flounder (wfAFP) is an alfa-helical, alanine-rich serum protein that helps protect against innoculative freezing from ice-laden seas. The AFP of a moth from the boreal forest, Choristoneura fumiferana (Cf), is a beta-helical threonine-rich protein that helps prevent freezing at the overwintering, caterpillar stage. In contrast, the beta-roll AFP from the grass, Lolium perenne (Lp), confers little freezing point depression and the plants readily freeze. Remarkably, AFPs also adsorb to tetrahyrofuran (THF) hydrate, changing the hydrate's octahedral morphology and, as well, inhibiting the growth of THF and gas hydrates. The hyperactive CfAFP, with 30-100 times the activity of wfAFP toward ice, showed far greater nucleation inhibition for THF hydrate than did a commercial hydrate inhibitor, poly(N-vinylpyrrolidone) (PVP). Active AFPs were also judged to be superior to PVP in that they inhibited the memory effect, a phenomenon whereby hydrate reforms at a faster rate soon after melting. An inactive mutant wfAFP, with an amino acid substitution at the ice-binding site, also reduced the growth of THF hydrate but was ineffective at suppressing hydrate reformation. These results suggest that the molecular properties important for ice adsorption and inhibition of hydrate reformation may be similar, and are distinct from those required for hydrate growth inhibition. The different AFPs also show markedly different aggregations on a third hydrophilic substrate, silica. Together these studies suggest that AFP adsorption to ice, hydrates and silica depends on the overall structure, specific residues and protein-protein interactions. (Author)

  1. Using Antifreeze Proteins to understand ice microstructure evolution (United States)

    Bayer-Giraldi, Maddalena; Azuma, Nobuhiko; Takata, Morimasa; Weikusat, Christian; Kondo, Hidemasa; Kipfstuhl, Sepp


    Polar ice sheets are considered a unique climate archive. The chemical analysis of its impurities and the development of its microstructure with depth give insight in past climate conditions as well as in the development of the ice sheet with time and deformation. Microstructural patterns like small grain size observed in specific depths are thought to be linked to the retarding effect of impurities on ice grain growth. Clear evidence of size or chemical composition of the impurities causing this effect is missing, but in this context a major role of nanoparticles has been suggested. In order to shed light on different mechanisms by which nanoparticles can control microstructure development we used antifreeze proteins (AFPs) as proxies for particles in ice. These proteins are small nanoparticles, approx. 5 nm in size, with the special characteristics of firmly binding to ice through several hydrogen bonds. We used AFPs from the sea-ice microalgae Fragilariopsis cylindrus (fcAFPs) in bubble-free, small-grained polycrystalline ice obtained by the phase-transition size refinement method. We explain how fcAFP bind to ice by presenting the 3-D-protein structure model inferred by X-ray structure analysis, and show the importance of the chemical interaction between particles and ice in controlling normal grain growth, comparing fcAFPs to other protein nanoparticles. We used modifications of fcAFPs for particle localization through fluorescence spectroscopy. Furthermore, the effect of fcAFPs on the driving factors for ice deformation during creep, i.e. on internal dislocations due to incorporation within the lattice and on the mobility of grain boundaries due to pinning, makes these proteins particularly interesting in studying the process of ice deformation.

  2. Expression and purification of sea raven type II antifreeze protein from Drosophila melanogaster S2 cells. (United States)

    Scotter, Andrew J; Kuntz, Douglas A; Saul, Michelle; Graham, Laurie A; Davies, Peter L; Rose, David R


    We present a system for the expression and purification of recombinant sea raven type II antifreeze protein, a cysteine-rich, C-type lectin-like globular protein that has proved to be a difficult target for recombinant expression and purification. The cDNAs encoding the pro- and mature forms of the sea raven protein were cloned into a modified pMT Drosophila expression vector. These constructs produced N-terminally His(6)-tagged pro- and mature forms of the type II antifreeze protein under the control of a metallothionein promoter when transfected into Drosophila melanogaster S2 cells. Upon induction of stable cell lines the two proteins were expressed at high levels and secreted into the medium. The proteins were then purified from the cell medium in a simple and rapid protocol using immobilized metal affinity chromatography and specific protease cleavage by tobacco etch virus protease. The proteins demonstrated antifreeze activity indistinguishable from that of wild-type sea raven antifreeze protein purified from serum as illustrated by ice affinity purification, ice crystal morphology, and their ability to inhibit ice crystal growth. This expression and purification system gave yields of 95 mg/L of fully active mature sea raven type II AFP and 9.6 mg/L of the proprotein. This surpasses all previous attempts to express this protein in Escherichia coli, baculovirus-infected fall armyworm cells and Pichia pastoris and will provide sufficient protein for structural analysis.

  3. Applications of type I antifreeze proteins: studies with model membranes & cryoprotectant properties. (United States)

    Inglis, Steven R; Turner, Jennifer J; Harding, Margaret M


    Antifreeze proteins (AFPs) and antifreeze glycoproteins (AFGPs), found in the body fluids of many species of polar fish allow them to survive in waters colder than the equilibrium freezing point of their blood and other internal fluids. Despite their structural diversity, all AF(G)Ps kinetically depress the temperature at which ice grows in a non-colligative manner and hence exhibit thermal hysteresis. AF(G)Ps also share the ability to interact with and protect mammalian cells and tissues from hypothermic damage (e.g., improved storage of human blood platelets at low temperatures), and are able to stabilize or disrupt membrane composition during low temperature and freezing stress (e.g., cryoprotectant properties in stabilization of sperm and oocytes). This review will summarize studies of AFPs with phospholipids and plant lipids, proposed mechanisms for inhibition of leakage from membranes, and cryoprotectant studies with biological samples. The major focus will be on the alpha-helical type I antifreeze proteins, and synthetic mutants, that have been most widely studied. For completeness, data on glycoproteins will also be presented. While a number of models to explain stabilization and destabilization of different lipid systems have been proposed, it is currently not possible to predict whether a particular AFP will stabilize or destabilize a given lipid system. Furthermore the relationship between the antifreeze property of thermal hysteresis and membrane stabilization is unknown. This lack of detailed knowledge about how AFPs function in the presence of different types of materials has hampered progress toward the development of antifreezes for cold storage of cells, tissues, and organs.

  4. A nonprotein thermal hysteresis-producing xylomannan antifreeze in the freeze-tolerant Alaskan beetle Upis ceramboides


    Walters, Kent R.; Serianni, Anthony S.; Sformo, Todd; Barnes, Brian M.; Duman, John G.


    Thermal hysteresis (TH), a difference between the melting and freezing points of a solution that is indicative of the presence of large-molecular-mass antifreezes (e.g., antifreeze proteins), has been described in animals, plants, bacteria, and fungi. Although all previously described TH-producing biomolecules are proteins, most thermal hysteresis factors (THFs) have not yet been structurally characterized, and none have been characterized from a freeze-tolerant animal. We isolated a highly a...

  5. Functional importance of short-range binding and long-range solvent interactions in helical antifreeze peptides. (United States)

    Ebbinghaus, Simon; Meister, Konrad; Prigozhin, Maxim B; Devries, Arthur L; Havenith, Martina; Dzubiella, Joachim; Gruebele, Martin


    Short-range ice binding and long-range solvent perturbation both have been implicated in the activity of antifreeze proteins and antifreeze glycoproteins. We study these two mechanisms for activity of winter flounder antifreeze peptide. Four mutants are characterized by freezing point hysteresis (activity), circular dichroism (secondary structure), Förster resonance energy transfer (end-to-end rigidity), molecular dynamics simulation (structure), and terahertz spectroscopy (long-range solvent perturbation). Our results show that the short-range model is sufficient to explain the activity of our mutants, but the long-range model provides a necessary condition for activity: the most active peptides in our data set all have an extended dynamical hydration shell. It appears that antifreeze proteins and antifreeze glycoproteins have reached different evolutionary solutions to the antifreeze problem, utilizing either a few precisely positioned OH groups or a large quantity of OH groups for ice binding, assisted by long-range solvent perturbation. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  6. HEAT INDUCIBLE EXPRESSION OF ANTIFREEZE PROTEIN GENES FROM THE BEETLES Tenebrio molitor AND Microdera punctipennis. (United States)

    Li, Jieqiong; Ma, Wenjing; Ma, Ji


    Antifreeze proteins (AFPs) play important roles in protecting poikilothermic organisms from cold damage. The expression of AFP genes (afps) is induced by low temperature. However, it is reported that heat can influence the expression of afps in the desert beetle Microdera punctipennis. To further detect whether heat also induce the expression of afps in other insects, and to determine the expression profiling of insect afps at different temperatures. The expression of antifreeze protein genes in the two beetles, Microdera punctipennis and Tenebrio molitor that have quite different living environment, under different temperatures were studied by using real-time quantitative PCR. Mild low temperatures (5~15 degree C), high temperature (38~47 degree C for M. punctipennis, or 37~42 degree C for T. molitor) and temperature difference (10~30 degree C) all stimulated strongly to the expression of AFP genes (Mpafps) in M. punctipennis which lives in the wild filed in desert. The mRNA level of Mpafps after M. punctipennis were exposed to these temperatures for 1h~5h was at least 30-fold of the control at 25 degree C. For T. molitor which is breeding in door with wheat bran all these temperatures stimulated significantly to the expression of Tmafps, while the extent and degree of the temperature stimulation on Tmafps expression were much lower than on Mpafps. After T. molitor were exposed to 5 degree C and 15 degree C for 1h~5h, the mRNA level of Tmafps was over 6-fold and 45-fold of the control at 25 degree C. High temperature (37~42 degree C) for 1h~3h treatments increased Tmafps mRNA level 4.8-fold of the control. Temperature difference of 10 degree C was effective in stimulating Tmafps expression. The expression of insect antifreeze protein genes both in M. punctipennis and T. molitor was induced by heat, suggesting that this phenomenon may be common in insects; the extent and degree of the influence differ in species that have different living conditions. The heat

  7. Marine Antifreeze Proteins: Structure, Function, and Application to Cryopreservation as a Potential Cryoprotectant

    Directory of Open Access Journals (Sweden)

    Hak Jun Kim


    Full Text Available Antifreeze proteins (AFPs are biological antifreezes with unique properties, including thermal hysteresis(TH,ice recrystallization inhibition(IRI,and interaction with membranes and/or membrane proteins. These properties have been utilized in the preservation of biological samples at low temperatures. Here, we review the structure and function of marine-derived AFPs, including moderately active fish AFPs and hyperactive polar AFPs. We also survey previous and current reports of cryopreservation using AFPs. Cryopreserved biological samples are relatively diverse ranging from diatoms and reproductive cells to embryos and organs. Cryopreserved biological samples mainly originate from mammals. Most cryopreservation trials using marine-derived AFPs have demonstrated that addition of AFPs can improve post-thaw viability regardless of freezing method (slow-freezing or vitrification, storage temperature, and types of biological sample type.

  8. Isolation of an antifreeze peptide from the Antarctic sponge Homaxinella balfourensis


    Wilkins, S. P.; Blum, A. J.; Burkepile, D. E.; Rutland, T. J.; Wierzbicki, A.; Kelly, M.; Hamann, M.T.


    Polar plants and animals survive in subzero waters (−2°C) and many of these marine organisms produce antifreeze proteins (AFPs) to better adapt themselves to these conditions. AFPs prevent the growth of ice crystals which disrupt cellular membranes and destroy cells by inhibiting crystallization of water within the organism. The hydrophilic extract of an Antarctic sponge Homaxinella balfourensis exhibited a non-colligative freezing point depression effect on the crystal morphology of water. T...

  9. De novo gene evolution of antifreeze glycoproteins in codfishes revealed by whole genome sequence data. (United States)

    Baalsrud, Helle Tessand; Tørresen, Ole Kristian; Hongrø Solbakken, Monica; Salzburger, Walter; Hanel, Reinhold; Jakobsen, Kjetill S; Jentoft, Sissel


    New genes can arise through duplication of a pre-existing gene or de novo from non-coding DNA, providing raw material for evolution of new functions in response to a changing environment. A prime example is the independent evolution of antifreeze glycoprotein genes (afgps) in the Arctic codfishes and Antarctic notothenioids to prevent freezing. However, the highly repetitive nature of these genes complicates studies of their organization. In notothenioids, afgps evolved from an extant gene, yet the evolutionary origin of afgps in codfishes is unknown. Here, we demonstrate that afgps in codfishes have evolved de novo from non-coding DNA 13-18 Ma, coinciding with the cooling of the Northern Hemisphere. Using whole-genome sequence data from several codfishes and notothenioids, we find higher copy number of afgp in species exposed to more severe freezing suggesting a gene dosage effect. Notably, antifreeze function is lost in one lineage of codfishes analogous to the afgp losses in non-Antarctic notothenioids. This indicates that selection can eliminate the antifreeze function when freezing is no longer imminent. Additionally, we show that evolution of afgp-assisting antifreeze potentiating protein genes (afpps) in notothenioids coincides with origin and lineage-specific losses of afgp. The origin of afgps in codfishes is one of the first examples of an essential gene born from non-coding DNA in a non-model species. Our study underlines the power of comparative genomics to uncover past molecular signatures of genome evolution, and further highlights the impact of de novo gene origin in response to a changing selection regime. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  10. Synthesis and characterization of natural and modified antifreeze glycopeptides: glycosylated foldamers. (United States)

    Nagel, Lilly; Plattner, Carolin; Budke, Carsten; Majer, Zsuzsanna; DeVries, Arthur L; Berkemeier, Thomas; Koop, Thomas; Sewald, Norbert


    In Arctic and Antarctic marine regions, where the temperature declines below the colligative freezing point of physiological fluids, efficient biological antifreeze agents are crucial for the survival of polar fish. One group of such agents is classified as antifreeze glycoproteins (AFGP) that usually consist of a varying number (n = 4-55) of [AAT]( n )-repeating units. The threonine side chain of each unit is glycosidically linked to β-D: -galactosyl-(1 → 3)-α-N-acetyl-D: -galactosamine. These biopolymers can be considered as biological antifreeze foldamers. A preparative route for stepwise synthesis of AFGP allows for efficient synthesis. The diglycosylated threonine building block was introduced into the peptide using microwave-enhanced solid phase synthesis. By this versatile solid phase approach, glycosylated peptides of varying sequences and lengths could be obtained. Conformational studies of the synthetic AFGP analogs were performed by circular dichroism experiments (CD). Furthermore, the foldamers were analysed microphysically according to their inhibiting effect on ice recrystallization and influence on the crystal habit.

  11. Cloning and expression of a novel antifreeze protein AFP72 from the beetle Tenebrio molitor. (United States)

    Yan, Qing-Hua; Yang, Li; Wang, Qing; Zhang, Hui-Rong; Shao, Qiang


    A novel antifreeze protein AFP72 cDNA (GenBbank accession No. AY929389) was obtained by RT-PCR from Tenebrio molitor. The 216 bp fragment encodes a protein of 72 amino acid residues. Sequence analysis revealed that the cDNA displays a high degree of homology with T. molitor antifreeze proteins, ranging up to 90.78%. Recombinant plasmids pMAL-p2X-afp72 and pMAL-c2X-afp72 were transferred into E. coil TBI to induce a MBP fusion protein by IPTG. The target fusion protein was released from the periplasm and cytoplasm by the cold osmotic shock procedure and sonication respectively. The content of the fusion protein came up to 38.9 and 41.5% of the total dissolved protein, respectively. The fusion protein was purified through an amylose affinity column, and incised by factor Xa. Molecular sieve chromatography was used to achieve a high state of purity of the target protein. The purified target protein displayed a single band in SDS-PAGE. The fusion protein was shown to increase resistance to low temperatures in bacteria. This finding could help in further investigations of the properties and function of antifreeze proteins.

  12. Interaction of Tenebrio Molitor Antifreeze Protein with Ice Crystal: Insights from Molecular Dynamics Simulations. (United States)

    Ramya, L; Ramakrishnan, Vigneshwar


    Antifreeze proteins (AFP) observed in cold-adapting organisms bind to ice crystals and prevent further ice growth. However, the molecular mechanism of AFP-ice binding and AFP-inhibited ice growth remains unclear. Here we report the interaction of the insect antifreeze protein (Tenebrio molitor, TmAFP) with ice crystal by molecular dynamics simulation studies. Two sets of simulations were carried out at 263 K by placing the protein near the primary prism plane (PP) and basal plane (BL) of the ice crystal. To delineate the effect of temperatures, both the PP and BL simulations were carried out at 253 K as well. The analyses revealed that the protein interacts strongly with the ice crystal in BL simulation than in PP simulation both at 263 K and 253 K. Further, it was observed that the interactions are primarily mediated through the interface waters. We also observed that as the temperature decreases, the interaction between the protein and the ice increases which can be attributed to the decreased flexibility and the increased structuring of the protein at low temperature. In essence, our study has shed light on the interaction mechanism between the TmAFP antifreeze protein and the ice crystal. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Animal ice-binding (antifreeze) proteins and glycolipids: an overview with emphasis on physiological function. (United States)

    Duman, John G


    Ice-binding proteins (IBPs) assist in subzero tolerance of multiple cold-tolerant organisms: animals, plants, fungi, bacteria etc. IBPs include: (1) antifreeze proteins (AFPs) with high thermal hysteresis antifreeze activity; (2) low thermal hysteresis IBPs; and (3) ice-nucleating proteins (INPs). Several structurally different IBPs have evolved, even within related taxa. Proteins that produce thermal hysteresis inhibit freezing by a non-colligative mechanism, whereby they adsorb onto ice crystals or ice-nucleating surfaces and prevent further growth. This lowers the so-called hysteretic freezing point below the normal equilibrium freezing/melting point, producing a difference between the two, termed thermal hysteresis. True AFPs with high thermal hysteresis are found in freeze-avoiding animals (those that must prevent freezing, as they die if frozen) especially marine fish, insects and other terrestrial arthropods where they function to prevent freezing at temperatures below those commonly experienced by the organism. Low thermal hysteresis IBPs are found in freeze-tolerant organisms (those able to survive extracellular freezing), and function to inhibit recrystallization - a potentially damaging process whereby larger ice crystals grow at the expense of smaller ones - and in some cases, prevent lethal propagation of extracellular ice into the cytoplasm. Ice-nucleator proteins inhibit supercooling and induce freezing in the extracellular fluid at high subzero temperatures in many freeze-tolerant species, thereby allowing them to control the location and temperature of ice nucleation, and the rate of ice growth. Numerous nuances to these functions have evolved. Antifreeze glycolipids with significant thermal hysteresis activity were recently identified in insects, frogs and plants. © 2015. Published by The Company of Biologists Ltd.

  14. An insight into the molecular basis for convergent evolution in fish antifreeze Proteins. (United States)

    Nath, Abhigyan; Chaube, Radha; Subbiah, Karthikeyan


    Antifreeze proteins (AFPs) prevent the growth of ice-crystals in order to enable certain organisms to survive under sub-zero temperature surroundings. These AFPs have evolved from different types of proteins without having any significant structural and sequence similarities among them. However, all the AFPs perform the same function of anti-freeze activity and are a classical example of convergent evolution. We have analyzed fish AFPs at the sequence level, the residue level and the physicochemical property group composition to discover molecular basis for this convergent evolution. Our study on amino acid distribution does not reveal any distinctive feature among AFPs, but comparative study of the AFPs with their close non-AFP homologs based on the physicochemical property group residues revealed some useful information. In particular (a) there is a similar pattern of avoidance and preference of amino acids in Fish AFP subtypes II, III and IV-Aromatic residues are avoided whereas small residues are preferred, (b) like other psychrophilic proteins, AFPs have a similar pattern of preference/avoidance for most of the residues except for Ile, Leu and Arg, and (c) most of the computed amino acids in preferred list are the key functional residues as obtained in previous predicted model of Doxey et al. For the first time this study revealed common patterns of avoidance/preference in fish AFP subtypes II, III and IV. These avoidance/preference lists can further facilitate the identification of key functional residues and can shed more light into the mechanism of antifreeze function. Copyright © 2013 Elsevier Ltd. All rights reserved.

  15. Antifreeze glycopeptides: from structure and activity studies to current approaches in chemical synthesis. (United States)

    Urbańczyk, Małgorzata; Góra, Jerzy; Latajka, Rafał; Sewald, Norbert


    Antifreeze glycopeptides (AFGPs) are a class of biological antifreeze agents found predominantly in Arctic and Antarctic species of fish. They possess the ability to regulate ice nucleation and ice crystal growth, thus creating viable life conditions at temperatures below the freezing point of body fluids. AFGPs usually consist of 4-55 repetitions of the tripeptide unit Ala-Ala-Thr that is O-glycosylated at the threonine side chains with β-D-galactosyl-(1 → 3)-α-N-acetyl-D-galactosamine. Due to their interesting properties and high antifreeze activity, they have many potential applications, e.g., in food industry and medicine. Current research is focused towards understanding the relationship between the structural preferences and the activity of the AFGPs, as well as developing time and cost efficient ways of synthesis of this class of molecules. Recent computational studies in conjunction with experimental results from NMR and THz spectroscopies were a possible breakthrough in understanding the mechanism of action of AFGPs. At the moment, as a result of these findings, the focus of research is shifted towards the analysis of behaviour of the hydration shell around AFGPs and the impact of water-dynamics retardation caused by AFGPs on ice crystal growth. In the field of organic synthesis of AFGP analogues, most of the novel protocols are centered around solid-phase peptide synthesis and multiple efforts are made to optimize this approach. In this review, we present the current state of knowledge regarding the structure and activity of AFGPs, as well as approaches to organic synthesis of these molecules with focus on the most recent developments.

  16. Perdeuteration, purification, crystallization and preliminary neutron diffraction of an ocean pout type III antifreeze protein

    International Nuclear Information System (INIS)

    Petit-Haertlein, Isabelle; Blakeley, Matthew P.; Howard, Eduardo; Hazemann, Isabelle; Mitschler, Andre; Haertlein, Michael; Podjarny, Alberto


    Perdeuterated type III antifreeze protein has been expressed, purified and crystallized. Preliminary neutron data collection showed diffraction to 1.85 Å resolution from a 0.13 mm 3 crystal. The highly homologous type III antifreeze protein (AFP) subfamily share the capability to inhibit ice growth at subzero temperatures. Extensive studies by X-ray crystallography have been conducted, mostly on AFPs from polar fishes. Although interactions between a defined flat ice-binding surface and a particular lattice plane of an ice crystal have now been identified, the fine structural features underlying the antifreeze mechanism still remain unclear owing to the intrinsic difficulty in identifying H atoms using X-ray diffraction data alone. Here, successful perdeuteration (i.e. complete deuteration) for neutron crystallographic studies of the North Atlantic ocean pout (Macrozoarces americanus) AFP in Escherichia coli high-density cell cultures is reported. The perdeuterated protein (AFP D) was expressed in inclusion bodies, refolded in deuterated buffer and purified by cation-exchange chromatography. Well shaped perdeuterated AFP D crystals have been grown in D 2 O by the sitting-drop method. Preliminary neutron Laue diffraction at 293 K using LADI-III at ILL showed that with a few exposures of 24 h a very low background and clear small spots up to a resolution of 1.85 Å were obtained using a ‘radically small’ perdeuterated AFP D crystal of dimensions 0.70 × 0.55 × 0.35 mm, corresponding to a volume of 0.13 mm 3

  17. Electro-Optical Properties Characterization of Fish Type III Antifreeze Protein


    Salvay, Andrés G.; Santos, Javier; Howard, Eduardo I.


    Antifreeze proteins (AFPs) are ice-binding proteins that depress the freezing point of water in a non-colligative manner without a significant modification of the melting point. Found in the blood and tissues of some organisms (such as fish, insects, plants, and soil bacteria), AFPs play an important role in subzero temperature survival. Fish Type III AFP is present in members of the subclass Zoarcoidei. AFPIII are small 7-kDa—or 14-kDa tandem—globular proteins. In the present work, we study ...


    Tanaka, Shunsuke; Takeichi, Kiyoshi; Masuyama, Yukiei; Takahashi, Naoto

    Snow and ice control in winter roads trends to be controlled by the skid friction coefficients in North America and North European countries at present, but the measurements are not necessarily easy. We studied on a simplified measurement method based on the relationship between skid friction coefficients and the bare pavement ratio (BPR) in the laboratory tests and field tests. The factors of BPR, surface textures and antifreezing materials which affect the skid friction coefficient are reviewed by a multiple linear regression analysis and a spectrum analysis, considering different freezing surfaces. These studies indicate that conclusions induced by laboratory tests could be applied to roads in service.

  19. An open source cryostage and software analysis method for detection of antifreeze activity

    DEFF Research Database (Denmark)

    Lørup Buch, Johannes; Ramløv, H


    The aim of this study is to provide the reader with a simple setup that can detect antifreeze proteins (AFP) by inhibition of ice recrystallisation in very small sample sizes. This includes an open source cryostage, a method for preparing and loading samples as well as a software analysis method....... The entire setup was tested using hyperactive AFP from the cerambycid beetle, Rhagium mordax. Samples containing AFP were compared to buffer samples, and the results are visualised as crystal radius evolution over time and in absolute change over 30 min. Statistical analysis showed that samples containing...

  20. Ice-surface adsorption enhanced colligative effect of antifreeze proteins in ice growth inhibition (United States)

    Mao, Yougang; Ba, Yong


    This Communication describes a mechanism to explain antifreeze protein's function to inhibit the growth of ice crystals. We propose that the adsorption of antifreeze protein (AFP) molecules on an ice surface induces a dense AFP-water layer, which can significantly decrease the mole fraction of the interfacial water and, thus, lower the temperature for a seed ice crystal to grow in a super-cooled AFP solution. This mechanism can also explain the nearly unchanged melting point for the ice crystal due to the AFP's ice-surface adsorption. A mathematical model combining the Langmuir theory of adsorption and the colligative effect of thermodynamics has been proposed to find the equilibrium constants of the ice-surface adsorptions, and the interfacial concentrations of AFPs through fitting the theoretical curves to the experimental thermal hysteresis data. This model has been demonstrated by using the experimental data of serial size-mutated beetle Tenebrio molitor (Tm) AFPs. It was found that the AFP's ice-surface adsorptions could increase the interfacial AFP's concentrations by 3 to 4 orders compared with those in the bulk AFP solutions.

  1. Agrobacterium-mediated transformation of modified antifreeze protein gene in strawberry

    Directory of Open Access Journals (Sweden)

    Srisulak Dheeranupattana


    Full Text Available The optimum condition for shoot regeneration from leaf explants of strawberry cultivar Tiogar was investigated. It was found that the best regeneration condition was MS medium containing N6-Benzyladenine (BA and 2,4-Dichlorophenoxy acetic acid (2,4-D at concentrations of 1 mg.l-1 and 0.2 mg.l-1, respectively. Antibiotics sensitivity test found that shoot regeneration from leaf explant was inhibited more than 90% at the concentration of kanamycin (Km as low as 5 mg.l-1. The modified gene encoding antifreeze protein isoform HPLC 6 was successfully constructed using codons which were optimally expressed in the strawberry plant. The antifreeze protein genes, naturally in plasmid pSW1 and modified in plasmid BB, were transformed to strawberry leaf explants by Agrobacterium tumefaciens LBA 4404. The strawberry plants, transformed with both AFP genes, were able to root in MS media containing 50 mg.l-1 Km, while no roots grew from nontransformed plant in this condition. Polymerase chain reaction indicated that the transgenes were integrated in the genome of transformants.

  2. An Effective Antifreeze Protein Predictor with Ensemble Classifiers and Comprehensive Sequence Descriptors

    Directory of Open Access Journals (Sweden)

    Runtao Yang


    Full Text Available Antifreeze proteins (AFPs play a pivotal role in the antifreeze effect of overwintering organisms. They have a wide range of applications in numerous fields, such as improving the production of crops and the quality of frozen foods. Accurate identification of AFPs may provide important clues to decipher the underlying mechanisms of AFPs in ice-binding and to facilitate the selection of the most appropriate AFPs for several applications. Based on an ensemble learning technique, this study proposes an AFP identification system called AFP-Ensemble. In this system, random forest classifiers are trained by different training subsets and then aggregated into a consensus classifier by majority voting. The resulting predictor yields a sensitivity of 0.892, a specificity of 0.940, an accuracy of 0.938 and a balanced accuracy of 0.916 on an independent dataset, which are far better than the results obtained by previous methods. These results reveal that AFP-Ensemble is an effective and promising predictor for large-scale determination of AFPs. The detailed feature analysis in this study may give useful insights into the molecular mechanisms of AFP-ice interactions and provide guidance for the related experimental validation. A web server has been designed to implement the proposed method.

  3. Operation of Kelvin Effect in the Activities of an Antifreeze Protein: A Molecular Dynamics Simulation Study. (United States)

    Midya, Uday Sankar; Bandyopadhyay, Sanjoy


    Ice growth and melting inhibition activities of antifreeze proteins (AFPs) are better explained by the adsorption-inhibition mechanism. Inhibition occurs as a result of the Kelvin effect induced by adsorbed protein molecules onto the surface of seed ice crystal. However, the Kelvin effect has not been explored by the state-of-the-art experimental techniques. In this work, atomistic molecular dynamics simulations have been carried out with Tenebrio molitor antifreeze protein ( TmAFP) placed at ice-water interface to probe the Kelvin effect in the mechanism of AFPs. Simulations show that, below equilibrium melting temperature, ice growth is inhibited through the convex ice-water interface formation toward the water phase and, above equilibrium melting temperature, ice melting is inhibited through the concave ice-water interface formation inward to ice phase. Simulations further reveal that the radius of curvature of the interface formed to stop the ice growth increases with decrease in the degree of supercooling. Our results are in qualitative agreement with the theoretical prediction of the Kelvin effect and thus reveal its operation in the activities of AFPs.

  4. Effects of preheating on ice growth in antifreeze polypeptides solutions in a narrow space (United States)

    Miyamoto, T.; Nishi, N.; Waku, T.; Tanaka, N.; Hagiwara, Y.


    We conducted measurements on the unidirectional freezing of aqueous solutions of polypeptide or of winter flounder antifreeze protein. The polypeptide was based on a part of the antifreeze protein. We measured temperatures in the solutions and ice with a small thermocouple, and defined the interface temperature as the temperature at the tip of the serrated or pectinate interface. It was found that the interface temperature of these solutions was lower than that of pure water. To vary the activity of these solutes, we preheated the solutions and cooled them before conducting the measurements. We found that preheating for several hours caused further decreases in the interface temperature and a decrease in the interface velocity. In addition, the inclined interfaces became wider as a result of the preheating. Thus, the supercooled states in the solutions were enhanced by the preheating. To investigate the reasons for these changes, we measured the aggregates of the solutes in the solutions. These aggregates became larger as a result of preheating. It can therefore be concluded that these large aggregates attenuated the ice growth by their interaction with the ice surfaces.

  5. Numerical prediction of micro-channel LD heat sink operated with antifreeze based on CFD method (United States)

    Liu, Gang; Liu, Yang; Wang, Chao; Wang, Wentao; Wang, Gang; Tang, Xiaojun


    To theoretically study the feasibility of antifreeze coolants applied as cooling fluids for high power LD heat sink, detailed Computational Fluid Dynamics (CFD) analysis of liquid cooled micro-channels heat sinks is presented. The performance operated with antifreeze coolant (ethylene glycol aqueous solution) compared with pure water are numerical calculated for the heat sinks with the same micro-channels structures. The maximum thermal resistance, total pressure loss (flow resistance), thermal resistance vs. flow-rate, and pressure loss vs. flow-rate etc. characteristics are numerical calculated. The results indicate that the type and temperature of coolants plays an important role on the performance of heat sinks. The whole thermal resistance and pressure loss of heat sinks increase significantly with antifreeze coolants compared with pure water mainly due to its relatively lower thermal conductivity and higher fluid viscosity. The thermal resistance and pressure loss are functions of the flow rate and operation temperature. Increasing of the coolant flow rate can reduce the thermal resistance of heat sinks; meanwhile increase the pressure loss significantly. The thermal resistance tends to a limit with increasing flow rate, while the pressure loss tends to increase exponentially with increasing flow rate. Low operation temperature chiefly increases the pressure loss rather than thermal resistance due to the remarkable increasing of fluid viscosity. The actual working point of the cooling circulation system can be determined on the basis of the pressure drop vs. flow rate curve for the micro-channel heat sink and that for the circulation system. In the same system, if the type or/and temperature of the coolant is changed, the working point is accordingly influenced, that is, working flow rate and pressure is changed simultaneously, due to which the heat sink performance is influenced. According to the numerical simulation results, if ethylene glycol aqueous

  6. Nanomaterials for efficiently lowering the freezing point of anti-freeze coolants. (United States)

    Hong, Haiping; Zheng, Yingsong; Roy, Walter


    In this paper, we report, for the first time, the effect of the lowered freezing point in a 50% water/50% anti-freeze coolant (PAC) or 50% water/50% ethylene glycol (EG) solution by the addition of carbon nanotubes and other particles. The experimental results indicated that the nano materials are much more efficient (hundreds fold) in lowering the freezing point than the regular ionic materials (e.g., NaCl). The possible explanation for this interesting phenomenon is the colligative property of fluid and relative small size of nano material. It is quite certain that the carbon nanotubes and metal oxide nano particles could be a wonderful candidate for the nano coolant application because they could not only increase the thermal conductivity, but also efficiently lower the freezing point of traditional coolants.

  7. Isolation of an antifreeze peptide from the Antarctic sponge Homaxinella balfourensis. (United States)

    Wilkins, S P; Blum, A J; Burkepile, D E; Rutland, T J; Wierzbicki, A; Kelly, M; Hamann, M T


    Polar plants and animals survive in subzero waters (-2 degrees C) and many of these marine organisms produce antifreeze proteins (AFPs) to better adapt themselves to these conditions. AFPs prevent the growth of ice crystals which disrupt cellular membranes and destroy cells by inhibiting crystallization of water within the organism. The hydrophilic extract of an Antarctic sponge Homaxinella balfourensis exhibited a non-colligative freezing point depression effect on the crystal morphology of water. The extract was purified by repeated reverse phase high-pressure liquid chromatography, then assayed and shown to contain several AFPs. The major peptide was isolated, analyzed using matrix-assisted laser desorption ionization mass spectrometry and the partial structure of the peptide identified through amino acid sequencing. AFPs have potential applications in agriculture, medicine and the food industry.

  8. Low thermodynamic but high kinetic stability of an antifreeze protein from Rhagium mordax

    DEFF Research Database (Denmark)

    Friis, Dennis Steven; Johnsen, Johannes Lørup; Kristiansen, Erlend


    , the RmAFP1 has only one disulfide bridge. The melting temperature, Tm, of the protein is determined to be 28.5°C (pH 7.4), which is much lower than most of those reported for AFPs or globular proteins in general. Despite its low melting temperature, both biophysical and activity measurements show......The equilibrium heat stability and the kinetic heat tolerance of a recombinant antifreeze protein (AFP) from the beetle Rhagium mordax (RmAFP1) are studied through differential scanning calorimetry and circular dichroism spectroscopy. In contrast to other insect AFPs studied with this respect...... that the protein almost completely refolds into the native state after repeated exposure of 70°C. RmAFP1 thus appears to be kinetically stable even far above its melting temperature. Thermodynamically, the insect AFPs seem to be dividable in three groups, relating to their content of disulfide bridges and widths...

  9. Re-evaluation of a bacterial antifreeze protein as an adhesin with ice-binding activity.

    Directory of Open Access Journals (Sweden)

    Shuaiqi Guo

    Full Text Available A novel role for antifreeze proteins (AFPs may reside in an exceptionally large 1.5-MDa adhesin isolated from an Antarctic Gram-negative bacterium, Marinomonas primoryensis. MpAFP was purified from bacterial lysates by ice adsorption and gel electrophoresis. We have previously reported that two highly repetitive sequences, region II (RII and region IV (RIV, divide MpAFP into five distinct regions, all of which require mM Ca(2+ levels for correct folding. Also, the antifreeze activity is confined to the 322-residue RIV, which forms a Ca(2+-bound beta-helix containing thirteen Repeats-In-Toxin (RTX-like repeats. RII accounts for approximately 90% of the mass of MpAFP and is made up of ∼120 tandem 104-residue repeats. Because these repeats are identical in DNA sequence, their number was estimated here by pulsed-field gel electrophoresis. Structural homology analysis by the Protein Homology/analogY Recognition Engine (Phyre2 server indicates that the 104-residue RII repeat adopts an immunoglobulin beta-sandwich fold that is typical of many secreted adhesion proteins. Additional RTX-like repeats in RV may serve as a non-cleavable signal sequence for the type I secretion pathway. Immunodetection shows both repeated regions are uniformly distributed over the cell surface. We suggest that the development of an AFP-like domain within this adhesin attached to the bacterial outer surface serves to transiently bind the host bacteria to ice. This association would keep the bacteria within the upper reaches of the water column where oxygen and nutrients are potentially more abundant. This novel envirotactic role would give AFPs a third function, after freeze avoidance and freeze tolerance: that of transiently binding an organism to ice.

  10. The mysteries of memory effect and its elimination with antifreeze proteins

    Energy Technology Data Exchange (ETDEWEB)

    Walker, V.; Gordienko, R.; Kuiper, M.; Huva, E.; Wu, Z. [Queen' s Univ., Kingston, ON (Canada). Dept. of Biology; Zeng, H.; Ripmeester, J. [Queen' s Univ., Kingston, ON (Canada). Dept. of Biology]|[National Research Council of Canada, Ottawa, ON (Canada). Steacie Inst. for Molecular Sciences


    With the decline in easily accessible and conventional hydrocarbon supplies, exploration will focus on hydrocarbons in deep offshore waters, in permafrost or in crystalline water as gas hydrates. Crystallization of water or water-encaged gas molecules takes place when nuclei reach a critical size, but the crystal growth may be inhibited by certain antifreeze proteins (AFPs). In this study, the authors hypothesized that the crystal lattice of gas hydrates may act as an alternative for substrate antifreeze proteins (AFPs). AFP-mediated inhibition of ice and clathrate hydrate crystallization was examined. Since the AFPs had a notable ability to eliminate the memory effect (ME) or the faster reformation of clathrate hydrates after melting, the authors were prompted to examine heterogeneous nucleation. Silica, served as a model nucleator hydrophilic surface. Quartz crystal microbalance-dissipation (QCM-D) experiments showed that an active AFP was tightly adsorbed to the silica surface. However, polyvinylpyrrolidone (PVP) and polyvinylcaprolactam (PVCap), 2 commercial hydrate kinetic inhibitors that do not eliminate ME, were not as tightly adsorbed. A mutant AFP inhibited tetrahydrofuran clathrate hydrate growth, but not ME. QCM-D analysis showed that adsorption of the mutant AFP was more similar to PVCap than the active AFP. It was concluded that although there is no evidence for memory in ice reformation, the crystallization of ice and hydrates, and the elimination of the more rapid recrystallization of hydrates, can be mediated by the same proteins. The properties of adsorbed layers can be effectively monitored by QCM-D. These study results provided useful information about the inhibition mechanism of heterogeneous nucleation of clathrate hydrate. The technique facilitates the screening of potential low dose hydrate inhibitors and residues in AFPs that are involved in silica adsorption. 24 refs., 1 tab., 4 figs.

  11. The effect of surface charge on the thermal stability and ice recrystallization inhibition activity of antifreeze protein III (AFP III). (United States)

    Deller, R C; Carter, B M; Zampetakis, I; Scarpa, F; Perriman, A W


    The aim of this study was to examine the effect of chemical cationization on the structure and function of antifreeze protein III (AFP III) over an extreme temperature range (-40°C to +90°C) using far-UV synchrotron radiation circular dichroism (SRCD) and ice recrystallization inhibition (IRI) assays. Chemical cationization was able to produce a modified AFP III with a net cationic charge at physiological pH that had enhanced resistance to denaturation at elevated temperatures, with no immediate negative impact on protein structure at subzero temperatures. Furthermore, cationized AFP III retained an IRI activity similar to that of native AFP III. Consequently, chemical cationization may provide a pathway to the development of more robust antifreeze proteins as supplementary cryoprotectants in the cryopreservation of clinically relevant cells. Copyright © 2017. Published by Elsevier Inc.

  12. Purification, crystal structure determination and functional characterization of type III antifreeze proteins from the European eelpout Zoarces viviparus

    DEFF Research Database (Denmark)

    Wilkens, Casper; Poulsen, Jens-Christian Navarro; Ramløv, Hans


    Antifreeze proteins (AFPs) are essential components of many organisms adaptation to cold temperatures. Fish type III AFPs are divided into two groups, SP isoforms being much less active than QAE1 isoforms. Two type III AFPs from Zoarces viviparus, a QAE1 (ZvAFP13) and an SP (ZvAFP6) isoform...... this is the first report of dimerization of AFP type III proteins....

  13. Influence of Block Copolymerization on the Antifreeze Protein Mimetic Ice Recrystallization Inhibition Activity of Poly(vinyl alcohol). (United States)

    Congdon, Thomas R; Notman, Rebecca; Gibson, Matthew I


    Antifreeze (glyco) proteins are produced by many cold-acclimatized species to enable them to survive subzero temperatures. These proteins have multiple macroscopic effects on ice crystal growth which makes them appealing for low-temperature applications-from cellular cryopreservation to food storage. Poly(vinyl alcohol) has remarkable ice recrystallization inhibition activity, but its mode of action is uncertain as is the extent at which it can be incorporated into other high-order structures. Here the synthesis and characterization of well-defined block copolymers containing poly(vinyl alcohol) and poly(vinylpyrrolidone) by RAFT/MADIX polymerization is reported, as new antifreeze protein mimetics. The effect of adding a large second hydrophilic block is studied across a range of compositions, and it is found to be a passive component in ice recrystallization inhibition assays, enabling retention of all activity. In the extreme case, a block copolymer with only 10% poly(vinyl alcohol) was found to retain all activity, where statistical copolymers of PVA lose all activity with very minor changes to composition. These findings present a new method to increase the complexity of antifreeze protein mimetic materials, while retaining activity, and also to help understand the underlying mechanisms of action.

  14. Growth-melt asymmetry in ice crystals under the influence of spruce budworm antifreeze protein

    Energy Technology Data Exchange (ETDEWEB)

    Pertaya, Natalya [Department of Physics and Astronomy, Ohio University, Athens, OH 45701 (United States); Celik, Yeliz [Department of Physics and Astronomy, Ohio University, Athens, OH 45701 (United States); DiPrinzio, Carlos L [Department of Physics and Astronomy, Ohio University, Athens, OH 45701 (United States); Wettlaufer, J S [Department of Geology and Geophysics, Yale University, New Haven, CT 06520-8109 (United States); Davies, Peter L [Department of Biochemistry, Queen' s University, Kingston, ON K7L 3N6 (Canada); Braslavsky, Ido [Department of Physics and Astronomy, Ohio University, Athens, OH 45701 (United States)


    Here we describe studies of the crystallization behavior of ice in an aqueous solution of spruce budworm antifreeze protein (sbwAFP) at atmospheric pressure. SbwAFP is an ice binding protein with high thermal hysteresis activity, which helps protect Choristoneura fumiferana (spruce budworm) larvae from freezing as they overwinter in the spruce and fir forests of the north eastern United States and Canada. Different types of ice binding proteins have been found in many other species. They have a wide range of applications in cryomedicine and cryopreservation, as well as the potential to protect plants and vegetables from frost damage through genetic engineering. However, there is much to learn regarding the mechanism of action of ice binding proteins. In our experiments, a solution containing sbwAFP was rapidly frozen and then melted back, thereby allowing us to produce small single crystals. These maintained their hexagonal shapes during cooling within the thermal hysteresis gap. Melt-growth-melt sequences in low concentrations of sbwAFP reveal the same shape transitions as are found in pure ice crystals at low temperature (-22 deg. C) and high pressure (2000 bar) (Cahoon et al 2006 Phys. Rev. Lett. 96 255502); while both growth and melt shapes display faceted hexagonal morphology, they are rotated 30 deg. relative to one another. Moreover, the initial melt shape and orientation is recovered in the sequence. To visualize the binding of sbwAFP to ice, we labeled the antifreeze protein with enhanced green fluorescent protein (eGFP) and observed the sbwAFP-GFP molecules directly on ice crystals using confocal microscopy. When cooling the ice crystals, facets form on the six primary prism planes (slowest growing planes) that are evenly decorated with sbwAFP-GFP. During melting, apparent facets form on secondary prism planes (fastest melting planes), leaving residual sbwAFP at the six corners of the hexagon. Thus, the same general growth-melt behavior of an apparently

  15. Fluorescence and confocal microscopy studies of the ice surface - antifreeze protein interactions. (United States)

    Pertaya, N.; Thomson, E.; Davies, P. L.; Braslavsky, I.


    Biomineralization is a phenomenon in which biological material influences mineral growth on the molecular level. A compelling example involves antifreeze proteins (AFPs) known to prevent fish and insects from freezing. AFPs have many potential applications in agriculture, biomedical science, and can be used as a model platform to understand biomineralization processes for future nanotechnology applications. Here we describe a new approach to study the interaction between AFPs and ice using fluorescence and confocal microscopy combined with a unique ice growth cell. After conjugating green fluorescent protein (GFP) to Type III AFP, we imaged the fluorescence signal around and inside of the ice crystals that emerged from the cooled AFP-GFP solution, and have observed an enhanced fluorescence signal at the edge of the ice crystal. In a second cell we observed a dramatic change in the ice growth morphology when AFPs were introduced into an initially pure system. Further developments of these methods will permit the direct imaging of the location and concentration of the AFPs on ice surfaces and enable a better understanding of their operation. Supported by CIHR, the Bosack and Kruger Foundation, Ohio and Yale Universities.

  16. FT-IR Spectra of Antifreeze Glycoproteins in Heavy Water and D2O Ice. (United States)

    Tsvetkova, N. M.; Crowe, J. H.; Feeney, R. H.; Fink, W. H.; Yeh, Yin


    This work presents FT-IR studies on the antifreeze glycoprotein (AFGP)/heavy water (D2O) mixtures during freezing and melting. AFGP in the blood serum of polar fish are known to prevent ice crystal growth by a non-colligative mechanism. There are 8 known fractions of AFGP (1 8) that range in molecular mass from 33.7 to 2.6 kD respectively, each composed of alanine-alanine-threonine repeats, with a disaccharide attached to the threonine residue. The smallest peptide (AFGP-8) is structurally different from fractions 1-5 in that it contains proline substituting for alanine in certain positions. Substantial linewidth change of the D20 bending mode (ca. 1210 cm-1) was measured with solutions containing fractions 2-5 during both freezing and thawing cycles, suggesting significant coupling between protein and water molecules. At the same time, the Amide I band between 1620 and 1675 cm-1 shows that 310 helix and random coils are the main conformations of fractions 2-5 and fraction 8 in the presence of ice. In liquid state, b-sheet dominates the secondary structure of AFGP 8, whereas b-sheet and random coil are the main conformations of AFGP 2-5. These results are discussed in terms of the ability of AFGP 2-5 to affect the surface states of ice.

  17. Electro-optical properties characterization of fish type III antifreeze protein. (United States)

    Salvay, Andrés G; Santos, Javier; Howard, Eduardo I


    Antifreeze proteins (AFPs) are ice-binding proteins that depress the freezing point of water in a non-colligative manner without a significant modification of the melting point. Found in the blood and tissues of some organisms (such as fish, insects, plants, and soil bacteria), AFPs play an important role in subzero temperature survival. Fish Type III AFP is present in members of the subclass Zoarcoidei. AFPIII are small 7-kDa-or 14-kDa tandem-globular proteins. In the present work, we study the behavior of several physical properties, such as the low-frequency dielectric permittivity spectrum, circular dichroism, and electrical conductivity of Fish Type III AFP solutions measured at different concentrations. The combination of the information obtained from these measurements could be explained through the formation of AFP molecular aggregates or, alternatively, by the existence of some other type of interparticle interactions. Thermal stability and electro-optical behavior, when proteins are dissolved in deuterated water, were also investigated.


    Directory of Open Access Journals (Sweden)

    Bhavana Sharma


    Full Text Available Antifreeze proteins (AFPs are a class of polypeptides which enables various organisms to survive subzero temperatures and have been found in vertebrates, invertebrates, plants, fungi and lichens. AFPs possess the characteristic thermal hysteresis (TH and ice recrystallization inhibition (IRI properties which allow them to adsorb the surface of ice crystals and inhibit their growth and recrystallization. AFPs are also known as ice restructuring proteins due to their ability to modify ice crystal morphology which leads to formation of hexagonal shape ice crystals in the presence of AFPs and disc shape AFPs in its absence. AFPs have various applications in medical, agricultural, industrial and biotechnological field. This review provides an overview of the AFPs, their TH and IRI properties and potential biotechnological applications of AFPs. Various conventional detection methods like Capillary assay and Differential Scanning Calorimetry (DSC with their advantages and disadvantages are discussed in detail along with the commonly used Splat assay and Nanoliter osmometer. Moreover, a novel, high-throughput and efficient nanobiotechnological method for AFP detection is also discussed. The method is based on colorimetric detection of freeze-labile gold nanoparticles and can provide an alternative to overcome the limitations of conventional methods by providing quick and easy way to screen AFPs in multiple systems simultaneously

  19. Balance between hydration enthalpy and entropy is important for ice binding surfaces in Antifreeze Proteins. (United States)

    Schauperl, Michael; Podewitz, Maren; Ortner, Teresa S; Waibl, Franz; Thoeny, Alexander; Loerting, Thomas; Liedl, Klaus R


    Antifreeze Proteins (AFPs) inhibit the growth of an ice crystal by binding to it. The detailed binding mechanism is, however, still not fully understood. We investigated three AFPs using Molecular Dynamics simulations in combination with Grid Inhomogeneous Solvation Theory, exploring their hydration thermodynamics. The observed enthalpic and entropic differences between the ice-binding sites and the inactive surface reveal key properties essential for proteins in order to bind ice: While entropic contributions are similar for all sites, the enthalpic gain for all ice-binding sites is lower than for the rest of the protein surface. In contrast to most of the recently published studies, our analyses show that enthalpic interactions are as important as an ice-like pre-ordering. Based on these observations, we propose a new, thermodynamically more refined mechanism of the ice recognition process showing that the appropriate balance between entropy and enthalpy facilitates ice-binding of proteins. Especially, high enthalpic interactions between the protein surface and water can hinder the ice-binding activity.

  20. Ancient climate change, antifreeze, and the evolutionary diversification of Antarctic fishes. (United States)

    Near, Thomas J; Dornburg, Alex; Kuhn, Kristen L; Eastman, Joseph T; Pennington, Jillian N; Patarnello, Tomaso; Zane, Lorenzo; Fernández, Daniel A; Jones, Christopher D


    The Southern Ocean around Antarctica is among the most rapidly warming regions on Earth, but has experienced episodic climate change during the past 40 million years. It remains unclear how ancient periods of climate change have shaped Antarctic biodiversity. The origin of antifreeze glycoproteins (AFGPs) in Antarctic notothenioid fishes has become a classic example of how the evolution of a key innovation in response to climate change can drive adaptive radiation. By using a time-calibrated molecular phylogeny of notothenioids and reconstructed paleoclimate, we demonstrate that the origin of AFGP occurred between 42 and 22 Ma, which includes a period of global cooling approximately 35 Ma. However, the most species-rich lineages diversified and evolved significant ecological differences at least 10 million years after the origin of AFGPs, during a second cooling event in the Late Miocene (11.6-5.3 Ma). This pattern indicates that AFGP was not the sole trigger of the notothenioid adaptive radiation. Instead, the bulk of the species richness and ecological diversity originated during the Late Miocene and into the Early Pliocene, a time coincident with the origin of polar conditions and increased ice activity in the Southern Ocean. Our results challenge the current understanding of the evolution of Antarctic notothenioids suggesting that the ecological opportunity that underlies this adaptive radiation is not linked to a single trait, but rather to a combination of freeze avoidance offered by AFGPs and subsequent exploitation of new habitats and open niches created by increased glacial and ice sheet activity.

  1. Calorimetric determination of inhibition of ice crystal growth by antifreeze protein in hydroxyethyl starch solutions. (United States)

    Hansen, T N; Carpenter, J F


    Differential scanning calorimetry and cryomicroscopy were used to investigate the effects of type I antifreeze protein (AFP) from winter flounder on 58% solutions of hydroxyethyl starch. The glass, devitrification, and melt transitions noted during rewarming were unaffected by 100 micrograms/ml AFP. Isothermal annealing experiments were undertaken to detect the effects of AFP-induced inhibition of ice crystal growth using calorimetry. A premelt endothermic peak was detected during warming after the annealing procedure. Increasing the duration or the temperature of the annealing for the temperature range from -28 and -18 degrees C resulted in a gradual increase in the enthalpy of the premelt endotherm. This transition was unaffected by 100 micrograms/ml AFP. Annealing between -18 and -10 degrees C resulted in a gradual decrease in the premelt peak enthalpy. This process was inhibited by 100 micrograms/ml AFP. Cryomicroscopic examination of the samples revealed that AFP inhibited ice recrystallization during isothermal annealing at -10 degrees C. Annealing at lower temperatures resulted in minimal ice recrystallization and no visible effect of AFP. Thus, the 100 micrograms/ml AFP to have a detectable influence on thermal events in the calorimeter, conditions must be used that result in significant ice growth without AFP and visible inhibition of this process by AFP. Images FIGURE 8 PMID:7690257

  2. Hydration behavior at the ice-binding surface of the Tenebrio molitor antifreeze protein. (United States)

    Midya, Uday Sankar; Bandyopadhyay, Sanjoy


    Molecular dynamics (MD) simulations have been carried out at two different temperatures (300 and 220 K) to study the conformational rigidity of the hyperactive Tenebrio molitor antifreeze protein (TmAFP) in aqueous medium and the structural arrangements of water molecules hydrating its surface. It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its nonice-binding surface (NIBS). The presence of a set of regularly arranged internally bound water molecules is found to play an important role in maintaining the flat rigid nature of the IBS. Importantly, the calculations reveal that the strategically located hydroxyl oxygens of the threonine (Thr) residues in the IBS influence the arrangements of five sets of ordered waters around it on two parallel planes that closely resemble the basal plane of ice. As a result, these waters can register well with the ice basal plane, thereby allowing the IBS to preferentially bind at the ice interface and inhibit its growth. This provides a possible molecular reason behind the ice-binding activity of TmAFP at the basal plane of ice.

  3. Reversible binding of the HPLC6 isoform of type I antifreeze proteins to ice surfaces and the antifreeze mechanism studied by multiple quantum filtering-spin exchange NMR experiment. (United States)

    Ba, Yong; Wongskhaluang, Jeff; Li, Jiabo


    Antifreeze proteins (AFPs) protect organisms from freezing damage by inhibiting the growth of seed-ice crystals. It has long been hypothesized that irreversible binding of AFPs to ice surfaces is responsible for inhibiting the growth of seed-ice crystals as such a mechanism supports the popularly accepted Kelvin effect for the explanation of local freezing-point depression. However, whether the binding is reversible or irreversible is still under debate due to the lack of direct experimental evidence. Here, we report the first direct experimental result, by using the newly developed multiple quantum (MQ) filtering-spin exchange NMR experiment, that shows that the binding of HPLC6 peptides to ice surfaces is reversible. It was found that the reversible process can be explained by the model of monolayer adsorption. These results suggest that the Kelvin effect is not suitable for explaining the antifreeze mechanism, and direct interactions between the peptides and the ice-surface binding sites are the driving forces for the binding of AFPs to ice surfaces. We propose that there exists a concentration gradient of AFP from an ice-binding surface to the solution due to the affinity of ice surfaces to AFPs. This concentration gradient creates a dense layer of AFP in contact with the ice-binding surface, which depresses the local freezing point because of the colligative property, but not the Kelvin effect.

  4. Revealing Surface Waters on an Antifreeze Protein by Fusion Protein Crystallography Combined with Molecular Dynamic Simulations. (United States)

    Sun, Tianjun; Gauthier, Sherry Y; Campbell, Robert L; Davies, Peter L


    Antifreeze proteins (AFPs) adsorb to ice through an extensive, flat, relatively hydrophobic surface. It has been suggested that this ice-binding site (IBS) organizes surface waters into an ice-like clathrate arrangement that matches and fuses to the quasi-liquid layer on the ice surface. On cooling, these waters join the ice lattice and freeze the AFP to its ligand. Evidence for the generality of this binding mechanism is limited because AFPs tend to crystallize with their IBS as a preferred protein-protein contact surface, which displaces some bound waters. Type III AFP is a 7 kDa globular protein with an IBS made up two adjacent surfaces. In the crystal structure of the most active isoform (QAE1), the part of the IBS that docks to the primary prism plane of ice is partially exposed to solvent and has clathrate waters present that match this plane of ice. The adjacent IBS, which matches the pyramidal plane of ice, is involved in protein-protein crystal contacts with few surface waters. Here we have changed the protein-protein contacts in the ice-binding region by crystallizing a fusion of QAE1 to maltose-binding protein. In this 1.9 Å structure, the IBS that fits the pyramidal plane of ice is exposed to solvent. By combining crystallography data with MD simulations, the surface waters on both sides of the IBS were revealed and match well with the target ice planes. The waters on the pyramidal plane IBS were loosely constrained, which might explain why other isoforms of type III AFP that lack the prism plane IBS are less active than QAE1. The AFP fusion crystallization method can potentially be used to force the exposure to solvent of the IBS on other AFPs to reveal the locations of key surface waters.

  5. Lateral transfer of a lectin-like antifreeze protein gene in fishes.

    Directory of Open Access Journals (Sweden)

    Laurie A Graham

    Full Text Available Fishes living in icy seawater are usually protected from freezing by endogenous antifreeze proteins (AFPs that bind to ice crystals and stop them from growing. The scattered distribution of five highly diverse AFP types across phylogenetically disparate fish species is puzzling. The appearance of radically different AFPs in closely related species has been attributed to the rapid, independent evolution of these proteins in response to natural selection caused by sea level glaciations within the last 20 million years. In at least one instance the same type of simple repetitive AFP has independently originated in two distant species by convergent evolution. But, the isolated occurrence of three very similar type II AFPs in three distantly related species (herring, smelt and sea raven cannot be explained by this mechanism. These globular, lectin-like AFPs have a unique disulfide-bonding pattern, and share up to 85% identity in their amino acid sequences, with regions of even higher identity in their genes. A thorough search of current databases failed to find a homolog in any other species with greater than 40% amino acid sequence identity. Consistent with this result, genomic Southern blots showed the lectin-like AFP gene was absent from all other fish species tested. The remarkable conservation of both intron and exon sequences, the lack of correlation between evolutionary distance and mutation rate, and the pattern of silent vs non-silent codon changes make it unlikely that the gene for this AFP pre-existed but was lost from most branches of the teleost radiation. We propose instead that lateral gene transfer has resulted in the occurrence of the type II AFPs in herring, smelt and sea raven and allowed these species to survive in an otherwise lethal niche.

  6. Testing antifreeze protein from the longhorn beetle Rhagium mordax as a kinetic gas hydrate inhibitor using a high-pressure micro differential scanning calorimeter

    DEFF Research Database (Denmark)

    Daraboina, Nagu; Perfeldt, Christine Malmos; von Solms, Nicolas


    pressure micro differential scanning calorimeter HP-mu DSC VII (Setaram Inc.) containing two 50 cc high pressure cells (maximum operating pressure 40 MPa; temperature range -40 to 120 degrees C) was employed to observe methane hydrate formation and decomposition in the presence of hyperactive antifreeze...

  7. Gold Nanoparticle Aggregation as a Probe of Antifreeze (Glyco) Protein-Inspired Ice Recrystallization Inhibition and Identification of New IRI Active Macromolecules. (United States)

    Mitchell, Daniel E; Congdon, Thomas; Rodger, Alison; Gibson, Matthew I


    Antifreeze (glyco)proteins are found in polar fish species and act to slow the rate of growth of ice crystals; a property known as ice recrystallization inhibition. The ability to slow ice growth is of huge technological importance especially in the cryopreservation of donor cells and tissue, but native antifreeze proteins are often not suitable, nor easily available. Therefore, the search for new materials that mimic this function is important, but currently limited by the low-throughout assays associated with the antifreeze properties. Here 30 nm gold nanoparticles are demonstrated to be useful colorimetric probes for ice recrystallization inhibition, giving a visible optical response and is compatible with 96 well plates for high-throughout studies. This method is faster, requires less infrastructure, and has easier interpretation than the currently used 'splat' methods. Using this method, a series of serum proteins were identified to have weak, but specific ice recrystallization inhibition activity, which was removed upon denaturation. It is hoped that high-throughput tools such as this will accelerate the discovery of new antifreeze mimics.

  8. Ice growth in supercooled solutions of a biological "antifreeze", AFGP 1-5: an explanation in terms of adsorption rate for the concentration dependence of the freezing point. (United States)

    Knight, C A; DeVries, A L


    It is widely accepted, and we agree, that the lowering of the temperature at which ice can grow in a water solution of one of the biological antifreezes is a result of adsorption of the antifreeze molecules at the ice surface. However, how this can produce a well-defined "freezing point" that varies with the solution concentration has remained problematical. The results of a series of measurements of ice growing in supercooled solutions of an effective antifreeze are reported and interpreted in terms of this fundamental problem. It seemed that the solution of the problem would have to rely upon adsorption rate, because that appeared to be the only way for the concentration in solution to be so important. The crystal growth results are most unusual, and appear to confirm this. The growth rates over a wide range of antifreeze concentration in solution (about 0.05 to 9 mg ml(-1)) are zero from the thermodynamic freezing point down to the "non-equilibrium" freezing point, where there is a very sudden increase to a plateau value that then remains about constant as the supercooling is increased by about 2 degrees C. The plateau values of growth rate are faster than those from pure water at the lower-supercooling ends of the plateaus, but slower at higher supercooling, until the growth rate starts rising toward that from pure water. These plateau values of growth rate increase markedly with increasing concentration of the antifreeze in solution. Along with these changes there are complex changes in the growth orientations, from c-axis spicules in the plateaus to those more characteristic of growth from pure water at greater supercooling. We conclude that the non-equilibrium freezing point is determined by the adsorption rate. It is the warmest temperature at which the ice growth rate on the basal plane (where the antifreeze does not adsorb) is fast enough to prevent the area of basal face on a growing ice crystal from becoming too small to grow, which is determined in

  9. Effects of anti-freeze concentration in the engine coolant on the cavitation temperature of a water pump

    International Nuclear Information System (INIS)

    David Huang, K.; Tzeng, S.-C.; Ma Weiping


    Improvements in engine-manufacturing technology have gradually increased the thermal efficiencies of engines as well as the burning temperature and pressure of fuels within the cylinders. Accordingly, greater heat dissipation are required. However, the volume of the radiators is constrained by the configuration of the engines, leading to excessive internal resistance in the engine-cooling system. Therefore, water pumps in engines are prone to cavitation, and air bubbles are likely to permeate into the anti-freeze, thereby severely reducing the performance, reliability and service life of the engines. Ethylene glycol (EG) is added to the radiator of some vehicles in cold areas to reduce the solidification point of the coolant and prevent freezing. This study probes the effects of the percentage of anti-freeze added to the cooling water in a water pump in an engine on the water-supply capability and cavitation temperature, whether air or burnt gas is present in the system. The results of this study have revealed that engines have a higher tolerance to air bubbles at lower rates of rotation. At a given fixed rotational speed, the tolerable cavitation temperature of an engine's water pump will fall slowly as the amount of air bubbles increases


    Directory of Open Access Journals (Sweden)

    Ricardo Vera Bravo


    Full Text Available A new strategy is presented for the designand synthesis of peptides that exhibitice-binding and antifreeze activity. Apennant-type dendrimer polypeptidescaffold combining an α-helical backbonewith four short β-strand branches wassynthesized in solid phase using Fmocchemistry in a divergent approach. The51-residue dendrimer was characterizedby reverse phase high performance liquidchromatography, mass spectrometry andcircular dichroism. Each β-strand branchcontained three overlapping TXT aminoacid repeats, an ice-binding motif foundin the ice-binding face of the sprucebudworm (Choristoneura fumiferanaand beetle (Tenebrio molitor antifreezeproteins. Ice crystals in the presence ofthe polypeptide monomer displayed flat,hexagonal plate morphology, similar tothat produced by weakly active antifreezeproteins. An oxidized dimeric form of thedendrimer polypeptide also produced flathexagonal ice crystals and was capableof inhibiting ice crystal growth upontemperature reduction, a phenomenontermed thermal hysteresis, a definingproperty of antifreeze proteins. Linkageof the pennant-type dendrimer to a trifunctionalcascade-type polypeptideproduced a trimeric macromolecule thatgave flat hexagonal ice crystals withhigher thermal hysteresis activity thanthe dimer or monomer and an ice crystal burst pattern similar to that producedby samples containing insect antifreezeproteins. This macromolecule was alsocapable of inhibiting ice recrystallization.

  11. Isolation and characterisation of sericin antifreeze peptides and molecular dynamics modelling of their ice-binding interaction. (United States)

    Wu, Jinhong; Rong, Yuzhi; Wang, Zhengwu; Zhou, Yanfu; Wang, Shaoyun; Zhao, Bo


    This study aimed to isolate and characterise a novel sericin antifreeze peptide and investigate its ice-binding molecular mechanism. The thermal hysteresis activity of ice-binding sericin peptides (I-SP) was measured and their activity reached as high as 0.94 °C. A P4 fraction, with high hypothermia protective activity and inhibition activity of ice recrystallisation, was obtained from I-SP, and a purified sericin peptide, named SM-AFP, with the sequence of TTSPTNVSTT and a molecular weight of 1009.50 Da was then isolated from the P4 fraction. Treatment of Lactobacillus delbrueckii Subsp. bulgaricus LB340 LYO with 100 μg/ml synthetic SM-AFP led to 1.4-fold increased survival (p Sericin peptides could be developed into beneficial cryoprotectants and used in frozen food processing. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Characterization of threonine side chain dynamics in an antifreeze protein using natural abundance {sup 13}C NMR spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Daley, Margaret E.; Sykes, Brian D. [University of Alberta, Department of Biochemistry, CIHR Group in Protein Structure and Function and Protein Engineering Network of Centres of Excellence (Canada)


    The dynamics of threonine side chains of the Tenebrio molitor antifreeze protein (TmAFP) were investigated using natural abundance {sup 13}C NMR. In TmAFP, the array of threonine residues on one face of the protein is responsible for conferring its ability to bind crystalline ice and inhibit its growth. Heteronuclear longitudinal and transverse relaxation rates and the {sup 1}H-{sup 13}C NOE were determined in this study. The C{alpha}H relaxation measurements were compared to the previously measured {sup 15}N backbone parameters and these are found to be in agreement. For the analysis of the threonine side chain motions, the model of restricted rotational diffusion about the {chi}{sub 1} dihedral angle was employed [London and Avitabile (1978) J. Am. Chem. Soc., 100, 7159-7165]. We demonstrate that the motion experienced by the ice binding threonine side chains is highly restricted, with an approximate upper limit of less than {+-}25 deg.

  13. Effects of three different types of antifreeze proteins on mouse ovarian tissue cryopreservation and transplantation.

    Directory of Open Access Journals (Sweden)

    Jaewang Lee

    Full Text Available Ovarian tissue (OT cryopreservation is effective in preserving fertility in cancer patients who have concerns about fertility loss due to cancer treatment. However, the damage incurred at different steps during the cryopreservation procedure may cause follicular depletion; hence, preventing chilling injury would help maintain ovarian function.This study was designed to investigate the beneficial effects of different antifreeze proteins (AFPs on mouse ovarian tissue cryopreservation and transplantation.Ovaries were obtained from 5-week-old B6D2F1 mice, and each ovary was cryopreserved using two-step vitrification and four-step warming procedures. In Experiment I, ovaries were randomly allocated into fresh, vitrification control, and nine experimental groups according to the AFP type (FfIBP, LeIBP, type III and concentration (0.1, 1, 10 mg/mL used. After vitrification and warming, 5,790 ovarian follicles were evaluated using histology and TUNEL assays, and immunofluorescence for τH2AX and Rad51 was used to detect DNA double-strand breaks (DSBs and repair (DDR, respectively. In Experiment II, 20 mice were randomly divided into two groups: one where the vitrification and warming media were supplemented with 10 mg/mL LeIBP, and the other where media alone were used (control. Ovaries were then autotransplanted under both kidney capsules 7 days after vitrification together with the addition of 10 mg/mL LeIBP in the vitrification-warming media. After transplantation, the ovarian follicles, the percentage of apoptotic follicles, the extent of the CD31-positive area, and the serum FSH levels of the transplanted groups were compared.In Experiment I, the percentage of total grade 1 follicles was significantly higher in the 10 mg/mL LeIBP group than in the vitrification control, while all AFP-treated groups had significantly improved grade 1 primordial follicle numbers compared with those of the vitrification control. The number of apoptotic (TUNEL

  14. Freeze resistance in rainbow smelt (Osmerus mordax): seasonal pattern of glycerol and antifreeze protein levels and liver enzyme activity associated with glycerol production. (United States)

    Lewis, Johanne M; Ewart, K Vanya; Driedzic, William R


    Rainbow smelt (Osmerus mordax) inhabit inshore waters along the North American Atlantic coast. During the winter, these waters are frequently ice covered and can reach temperatures as low as -1.9 degrees C. To prevent freezing, smelt accumulate high levels of glycerol, which lower the freezing point via colligative means, and antifreeze proteins (AFP). The up-regulation of the antifreeze response (both glycerol and AFP) occurs in early fall, when water temperatures are 5 degrees -6 degrees C. The accumulation of glycerol appears to be the main mechanism of freeze resistance in smelt because it contributes more to the lowering of the body's freezing point than the activity of the AFP (0.5 degrees C vs. 0.25 degrees C for glycerol and AFP, respectively) at a water temperature of -1.5 degrees C. Moreover, AFP in smelt appears to be a safeguard mechanism to prevent freezing when glycerol levels are low. Significant increases in activities of the liver enzymes glycerol 3-phosphate dehydrogenase (GPDH), alanine aminotransferase (AlaAT), and phosphoenolpyruvate carboxykinase (PEPCK) during the initiation of glycerol production and significant correlations between enzyme activities and plasma glycerol levels suggest that these enzymes are closely associated with the synthesis and maintenance of elevated glycerol levels for use as an antifreeze. These findings add further support to the concept that carbon for glycerol is derived from amino acids.

  15. Aqueous geochemistry in icy world interiors: Equilibrium fluid, rock, and gas compositions, and fate of antifreezes and radionuclides (United States)

    Neveu, Marc; Desch, Steven J.; Castillo-Rogez, Julie C.


    The geophysical evolution of many icy moons and dwarf planets seems to have provided opportunities for interaction between liquid water and rock (silicate and organic solids). Here, we explore two ways by which water-rock interaction can feed back on geophysical evolution: the production or consumption of antifreeze compounds, which affect the persistence and abundance of cold liquid; and the potential leaching into the fluid of lithophile radionuclides, affecting the distribution of a long-term heat source. We compile, validate, and use a numerical model, implemented with the PHREEQC code, of the interaction of chondritic rock with pure water and with C, N, S-bearing cometary fluid, thought to be the materials initially accreted by icy worlds, and describe the resulting equilibrium fluid and rock assemblages at temperatures, pressures, and water-to-rock ratios of 0-200 ° C, 1-1000 bar, and 0.1-10 by mass, respectively. Our findings suggest that water-rock interaction can strongly alter the nature and amount of antifreezes, resulting in solutions rich in reduced nitrogen and carbon, and sometimes dissolved H2, with additional sodium, calcium, chlorine, and/or oxidized carbon. Such fluids can remain partially liquid down to 176 K if NH3 is present. The prominence of Cl in solution seems to hinge on its primordial supply in ices, which is unconstrained by the meteoritical record. Equilibrium assemblages, rich in serpentine and saponite clays, retain thorium and uranium radionuclides unless U-Cl or U-HCO3 complexing, which was not modeled, significantly enhances U solubility. However, the radionuclide 40 K can be leached at high water:rock ratio and/or low temperature at which K is exchanged with ammonium in minerals. We recommend the inclusion of these effects in future models of the geophysical evolution of ocean-bearing icy worlds. Our simulation products match observations of chloride salts on Europa and Enceladus; CI chondrites mineralogies; the observation of

  16. Relationship of amino acid composition and molecular weight of antifreeze glycopeptides to non-colligative freezing point depression. (United States)

    Schrag, J D; O'Grady, S M; DeVries, A L


    Many polar fishes synthesize a group of eight glycopeptides that exhibit a non-colligative lowering of the freezing point of water. These glycopeptides range in molecular weight between 2600 and 33 700. The largest glycopeptides [1-5] lower the freezing point more than the small ones on a weight basis and contain only two amino acids, alanine and threonine, with the disaccharide galactose-N-acetyl-galactosamine attached to threonine. The small glycopeptides, 6, 7, and 8, also lower the freezing point and contain proline, which periodically substitutes for alanine. Glycopeptides with similar antifreeze properties isolated from the saffron cod and the Atlantic tomcod contain an additional amino acid, arginine, which substitutes for threonine in glycopeptide 6. In this study we address the question of whether differences in amino acid composition or molecular weight between large and small glycopeptides are responsible for the reduced freezing point depressing capability of the low molecular weight glycopeptides. The results indicate that the degree of amino acid substitutions that occur in glycopeptides 6-8 do not have a significant effect on the unusual freezing point lowering and that the observed decrease in freezing point depression with smaller glycopeptides can be accounted for on the basis of molecular weight.

  17. The role of endogenous antifreeze protein enhancers in the hemolymph thermal hysteresis activity of the beetle Dendroides canadensis. (United States)

    Duman, John G.; Serianni, Anthony S.


    Antifreeze proteins (AFPs) lower the freezing point of water by a non-colligative mechanism, but do not lower the melting point, therefore producing a difference between the freezing and melting points termed thermal hysteresis. Thermal hysteresis activity (THA) of AFPs from overwintering larvae of the beetle Dendroides canadensis is dependent upon AFP concentration and the presence of enhancers of THA which may be either other proteins or low molecular mass enhancers. The purpose of this study was to determine the relative contributions of endogenous enhancers in winter D. canadensis hemolymph.Winter hemolymph collected over four successive winters (1997-1998 to 2000-2001) was tested. The first three of these winters were the warmest on record in this area, while December of the final year was the coldest on record. Protein and low molecular mass enhancers raised hemolymph THA 60-97% and 35-55%, respectively, based on hemolymph with peak THA for each year collected over the four successive winters. However, the hemolymph AFPs were not maximally enhanced since addition of the potent enhancer citrate (at non-physiologically high levels) resulted in large increases in THA. (13)NMR showed that glycerol was the only low molecular mass solute present in sufficiently high concentrations in the hemolymph to function as an enhancer. Maximum THA appears to be approximately 8.5 degrees C.

  18. Oscillations and accelerations of ice crystal growth rates in microgravity in presence of antifreeze glycoprotein impurity in supercooled water (United States)

    Furukawa, Yoshinori; Nagashima, Ken; Nakatsubo, Shun-Ichi; Yoshizaki, Izumi; Tamaru, Haruka; Shimaoka, Taro; Sone, Takehiko; Yokoyama, Etsuro; Zepeda, Salvador; Terasawa, Takanori; Asakawa, Harutoshi; Murata, Ken-Ichiro; Sazaki, Gen


    The free growth of ice crystals in supercooled bulk water containing an impurity of glycoprotein, a bio-macromolecule that functions as ‘antifreeze’ in living organisms in a subzero environment, was observed under microgravity conditions on the International Space Station. We observed the acceleration and oscillation of the normal growth rates as a result of the interfacial adsorption of these protein molecules, which is a newly discovered impurity effect for crystal growth. As the convection caused by gravity may mitigate or modify this effect, secure observations of this effect were first made possible by continuous measurements of normal growth rates under long-term microgravity condition realized only in the spacecraft. Our findings will lead to a better understanding of a novel kinetic process for growth oscillation in relation to growth promotion due to the adsorption of protein molecules and will shed light on the role that crystal growth kinetics has in the onset of the mysterious antifreeze effect in living organisms, namely, how this protein may prevent fish freezing.

  19. Effects of short-time preheating on ice growth in antifreeze polypeptides solutions in a narrow space (United States)

    Miyamoto, T.; Nishi, N.; Waku, T.; Tanaka, N.; Hagiwara, Y.


    We conducted experiments on the unidirectional freezing of solutions of winter flounder antifreeze protein or of a polypeptide which was based on twelve amino-acid residues of this protein. The temperature in the solutions and ice was measured with a small thermocouple. The interface temperature was defined as the temperature at the tip of the serrated or pectinate interface. The interface temperature of these solutions was lower than that of pure water. To vary this supercooling activity of these solutes, we preheated the solutions and cooled them before conducting identical experiments. It was found that short-time preheating caused further decreases in the interface temperature and interface velocities. Furthermore, the inclined interfaces and the narrow liquid regions inside the ice area became wider. To investigate the reasons for these changes, we measured aggregates of the solutes in the solutions. These aggregates were found to become larger as a result of preheating. Thus, it can be concluded that these large aggregates attenuated the ice growth by their interaction with ice. Finally, we carried out similar measurements by using pH-adjusted solutions of the protein to produce aggregates without preheating, and obtained similar supercooling enhancement by the aggregates. Thus, the effects of thermal denaturation on the supercooling were not significant in the preheating.

  20. Crystallization and preliminary X-ray crystallographic analysis of Ca{sup 2+}-independent and Ca{sup 2+}-dependent species of the type II antifreeze protein

    Energy Technology Data Exchange (ETDEWEB)

    Nishimiya, Yoshiyuki; Kondo, Hidemasa [Functional Protein Research Group, Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo 062-8517 (Japan); Yasui, Masanori [Functional Protein Research Group, Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo 062-8517 (Japan); Division of Biological Sciences, Graduate School of Science, Hokkaido University, N8W5, Kita, Sapporo 060-0808 (Japan); Sugimoto, Hiroshi [Biometal Science Laboratory, Riken SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo, Hyogo 679-5148 (Japan); Noro, Natsuko; Sato, Ryoko [Functional Protein Research Group, Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo 062-8517 (Japan); Suzuki, Mamoru [Insititute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Miura, Ai [Functional Protein Research Group, Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo 062-8517 (Japan); Tsuda, Sakae, E-mail: [Functional Protein Research Group, Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo 062-8517 (Japan); Division of Biological Sciences, Graduate School of Science, Hokkaido University, N8W5, Kita, Sapporo 060-0808 (Japan)


    Crystallization and X-ray data analyses were successful for both Ca{sup 2+}-independent and Ca{sup 2+}-dependent species of the type II antifreeze protein. The resolution of the crystal was 1.35 Å for the Ca{sup 2+}-independent species, and was 1.25 and 1.06 Å for the Ca{sup 2+}-dependent species in the Ca{sup 2+}-free and -bound states, respectively. Ca{sup 2+}-independent and Ca{sup 2+}-dependent species of the type II antifreeze protein (AFP) were both crystallized using the hanging-drop vapour-diffusion method. It appeared that the crystal of the Ca{sup 2+}-independent species from Brachyosis rostratus belongs to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 43.3, b = 48.4, c = 59.7 Å, and diffraction data were collected to 1.34 Å resolution. For the Ca{sup 2+}-dependent type II AFP species from Hypomesus nipponensis, crystallization was carried out for its Ca{sup 2+}-free and Ca{sup 2+}-bound states. 1.25 Å resolution data were collected from the crystal in the Ca{sup 2+}-free state, which exhibited P3{sub 1}21 (or P3{sub 2}21) symmetry, with unit-cell parameters a = b = 66.0, c = 50.3 Å. Data collection could be extended to 1.06 Å resolution for the crystal in the Ca{sup 2+} -bound state, which appeared to be isomorphous to the crystal in the Ca{sup 2+}-free state (unit-cell parameters a = b = 66.0, c = 49.8 Å). These data will allow us to determine the high-resolution structures of the two species of type II AFP.

  1. Stable transmission and transcription of newfoundland ocean pout type III fish antifreeze protein (AFP) gene in transgenic mice and hypothermic storage of transgenic ovary and testis. (United States)

    Bagis, Haydar; Aktoprakligil, Digdem; Mercan, Hande Odaman; Yurdusev, Nevzat; Turgut, Gazi; Sekmen, Sakir; Arat, Sezen; Cetin, Seyfettin


    Here we describe the generation of transgenic mice carrying type III fish antifreeze protein (AFP) gene and evaluate whether AFP type III protects transgenic mouse ovaries and testes from hypothermic storage. AFPs exist in many different organisms. In fish, AFPs protect the host from freezing at temperatures below the colligative freezing point by adsorbing to the surface of nucleating ice crystals and inhibiting their growth. The transgenic expression of AFP holds great promise for conferring freeze-resistant plant and animal species. AFP also exhibits a potential for the cryopreservation of tissues and cells. In this study, we have generated 42 founder mice harboring the Newfoundland ocean pout (OP5A) type III AFP transgene and established one transgenic line (the line #6). This study demonstrated that AFP gene construct has been stably transmitted to the mouse progeny in the F3 generations in the line #6. Furthermore, the presence of AFP transcripts was confirmed by RT-PCR analysis on cDNAs from liver, kidney, ovarian, and testis tissues of the mouse from F3 generation in this line. These results indicate that ocean pout type III AFP gene could be integrated and transmitted to the next generation and stably transcribed in transgenic mice. In histological analysis of testis and ovarian tissues of nontransgenic control and AFP transgenic mice it has been shown that both tissues of AFP transgenic mice were protected from hypothermic storage (+4 degrees C). The AFP III transgenic mice obtained for the first time in this study would be useful for investigating the biological functions of AFP in mammalian systems and also its potential role in cryopreservation.

  2. Characterization of type IV antifreeze gene in Nile tilapia (Oreochromis niloticus) and influence of cold and hot weather on its expression and some immune-related genes. (United States)

    Ammar, Asmma Y; El Nahas, Abeer F; Mahmoud, Shawky; Barakat, Mohamed E; Hassan, Asmaa M


    The aim of this work is to study the effect of the thermal stress of ambient temperature during winter and summer on the expression of type IV antifreeze gene (ANF IV) in different tissues of Nile tilapia (Oreochromis niloticus) as well as some immune-related genes. At first, genomic ANF IV gene was characterized from one fish; 124 amino acids were identified with 92.7% similarity with that on the gene bank. Expression of ANF IV and immune-related genes were done twice, once at the end of December (winter sample, temperature 14 °C) and the other at August (summer sample, temperature 36 °C). Assessment of ANF IV gene expression in different organs of fish was done; splenic mRNA was used for assessment of immune-related gene transcripts (CXCl2 chemokine, cc-chemokine, INF-3A, and MHC IIβ). Winter expression analysis of AFP IV in O. niloticus revealed significant upregulation of mRNA transcript levels in the intestine, gills, skin, spleen, liver, and brain with 324.03-, 170.06-, 107.63-, 97.61-, 94.35-, and 27.85-folds, respectively. Furthermore, upregulation in the gene was observed in some organs during summer: in the liver, gills, skin, intestine, and brain with lower levels compared with winter. The level of expression of immune-related genes in winter is significantly higher than summer in all assessed genes. Cc-chemokine gene expression was the most affected in both winter and summer. Variable expression profile of ANF IV in different organs and in different seasons together with its amino acid similarity of N-terminal and C-terminal with apolipoprotein (lipid binder) and form of high-density lipoprotein (HDL) suggests a different role for this protein which may be related to lipid metabolism.

  3. Improved viability and reduced apoptosis in sub-zero 21-hour preservation of transplanted rat hearts using anti-freeze proteins. (United States)

    Amir, Gabriel; Rubinsky, Boris; Basheer, Sheick Yousif; Horowitz, Liana; Jonathan, Leor; Feinberg, Micha S; Smolinsky, Aram K; Lavee, Jacob


    Freeze-tolerant fish survive sub-zero temperatures by non-colligatively lowering the freezing temperature of their body fluids using anti-freeze proteins (AFPs). We sought to evaluate and compare the effects of prolonged sub-zero cryopreservation of transplanted rat hearts using AFP I or AFP III. Two heterotopic rat heart transplantation protocols were used. In Protocol 1 (n = 104), hearts (n = 8/group) were preserved for 12, 18 and 24 hours in University of Wisconsin solution (UW) at 4 degrees C, UW at -1.3 degrees C, UW/AFP I at -1.3 degrees C and UW/AFP III at -1.3 degrees C, with and without nucleation. Post-operative evaluation consisted of visual viability scoring of the hearts after 60 minutes. Protocol 2 (n = 58) involved evaluation of 24-hour post-transplant viability, echocardiography (fractional shortening [FS], left ventricular end-systolic and -diastolic diameter [ESD, EDD] and anterior and posterior wall systolic and diastolic thickness [AWT-S, AWT-D, PWT-S, PWT-D]), TUNEL staining and electron microscopy (EM) findings for hearts preserved for 18, 21 and 24 hours in UW at 4 degrees C or UW/AFP III at -1.3 degrees C. Hearts preserved in UW at -1.3 degrees C with nucleation froze and died. Three of 8 hearts preserved in UW at 4 degrees C for 24 hours died, whereas all hearts preserved at -1.3 degrees C survived. Hearts preserved in UW/AFP for 18 and 24 hours at -1.3 degrees C had superior viability scores compared with those in UW at 4 degrees C. Hearts in AFP III at -1.3 degrees C displayed greater AWT-S and AWT-D (3.5 +/- 0.2 vs 2.4 +/- 0.2, p hour preservation. In the 21-hour preservation group, AFP-treated hearts displayed improved echocardiographic systolic contraction indices, including: improved FS (27 +/- 3.7 vs 15 +/- 4, p = 0.04); diminished ESD (0.28 +/- 0.57 vs 0.47 +/- 0.6, p zero cryopreservation, AFPs protect the heart from freezing, improve survival and hemodynamics, and reduce apoptotic cell death.

  4. Anti-freeze proteins: characteristics, mechanism of action and applications

    NARCIS (Netherlands)

    Stolle-Smits, T.; Kreuwels, M.J.J.; Wichers, H.J.


    Antivrieseiwitten hebben de eigenschap om het vriespunt te verlagen en rekristallisatie van ijs tegen te gaan. Deze eiwitten komen voor in poolvissen, insekten, planten en micro-organismen. Bovendien geven ze de ijskristallen een andere morfologie

  5. Antifreeze proteins enable plants to survive in freezing conditions

    Indian Academy of Sciences (India)

    1. Acaenamagellanica. Prickly burr. Doucet et al. 2000. 2. Acer saccharoides. Maple. Doucet et al. 2000. 3. Agrostistenuis. Creeping bentgrass. Doucet et al. 2000. 4. Alliarapetiolata. Garlic mustard. Urrutia et al. 1992. 5. Ammopiptanthusmongolicus. Evergreen legume. Wang et al. 2003. 6. Aster cordifolius. Wood aster.

  6. In silico characterization of antifreeze proteins using computational ...

    Indian Academy of Sciences (India)


    organs.12 Chao et al13 have reported the relative effi- cacy of AFP types I, II and III in protecting the red blood cells. Numerous structure and function studies ..... Table 7. PDB templates (first 2 hits with maximum % identity) obtained using BLASTP search against the Pro- tein Data Bank. Accession number. PDB code.

  7. Studies on new antifreeze protein from the psychrophilic diatom ...

    African Journals Online (AJOL)



    Sep 12, 2011 ... forward primer and XhoI to reverse primer for cloning into pET vector. Bacterial and plant expressions. PCR products were analyzed and resolved on 1% agarose gel. (w/v). Bands with expected lengths were excised from the gel with razor blade and purified using the QIAquick gel extraction kit. (Qiagen, CA ...

  8. Atmospheric wet-type cooling tower with antifreeze system

    International Nuclear Information System (INIS)

    Coic, P.


    The cooling tower has air inlets at its base, a network of pipes which distributes the air to be cooled above the packing, and valves to isolate a part of the network. It includes also a bypass circuit, provided with means to control the flow rate fraction which is by-passed [fr

  9. In silico characterization of antifreeze proteins using computational ...

    Indian Academy of Sciences (India)


    GRAVY, Grand Average Hydropathy. structure of protein (3D coordinates data). The 3D structure of AFPs Q01758 and P05140 were gener- ated by homology modelling using Esypred34 server. The similar 3D structures (for the AFPs Q01758 and. P05140 sequences) in the Protein Data bank. ( were identified ...

  10. Antifreeze proteins enable plants to survive in freezing conditions

    Indian Academy of Sciences (India)

    Common name. References. 28. Hordeumvulgare. Barley. Antikainen and Griffith 1997. 29. Hydrophyllumvirginianum. Virginia waterleaf. Urrutia et al. 1992. 30. Juncussquarrosus. Heath rush. Doucet et al. 2000. 31. Loliumperenne. Perennial ryegrass. Sidebottom et al. 2000; Pudney et al. 2003. 32. Northofagusantarctica.

  11. Structure/Function Studies of Insect Antifreeze Proteins

    National Research Council Canada - National Science Library

    Duman, John


    ...-x5-x6-Cys-X8-X9-Ala-X11-Th-X13 where X3 and X1 tend toward charged residues, X5 toward threonine or serine, X9 toward asparagine or aspartate, X6 toward asparagine or lysine, and X13 toward alanine...

  12. Anti-freeze coatings for the rotor blades of wind turbines; Anti-freeze Beschichtungen fuer Rotorblaetter von Windenergieanlagen

    Energy Technology Data Exchange (ETDEWEB)

    Siegmann, K.; Kaufmann, A.; Hirayama, M.


    This final report for the Swiss Federal Office of Energy (SFOE) takes a look at projects involving the development of suggestions for coatings for the rotor blades of wind turbines. The coatings are to reduce the formation of hoarfrost on the leading edges of the blades. Various coatings are described and the mechanisms involved in the formation of the frost and in keeping the blades as free as possible from frost are discussed. Global know-how on the subject is discussed, as is know-how available in Europe and Switzerland. Manufacturers, planning offices and installation operators are listed, as are research institutes who are dealing with this problem. In the summary, the authors stress the importance of choosing the coating most suitable for the actual climatic conditions at the wind turbine's location. A suggestion is made for further work in this area.

  13. Detecting seasonal variation of antifreeze protein distribution in Rhagium mordax using immunofluorescence and high resolution microscopy

    DEFF Research Database (Denmark)

    Buch, Johannes Lørup; Ramløv, Hans


    with UV reflected light microscopy. An automated software analysis method was developed in order to discard autofluorescence, and quantify fluorescence from bound antibodies. The results show that R. mordax cuticle and gut exhibit a higher degree of fluorophore-bound fluorescence during summer, than...

  14. Antifreeze Production and Cold-Tolerance in Overwintering Purple Martin Fleas, Ceratophyllus idius Jordan and Rothschild. (United States)


    rates for females than for males, and better survival for both sexes at -5 or -6 C than under ambient conditions. A survey of the literature indicates...collembolans and oribatid mites from windswept mountain ridges. Oikos 29: 118-126. Storey, J.M.; Storey, K.B. 1983a. Regulation of cryo- protectant

  15. Additifs anticongelants pour huiles de moteurs Antifreezing Additives for Motor Oils

    Directory of Open Access Journals (Sweden)

    Denis J.


    Full Text Available Cet article présente d'abord les principales familles chimiques d'additifs de congélation. Les paraffines normales restant dans l'huile après le déparaffinage cristallisent à basse température sous forme de fines lamelles constituées de plusieurs couches de molécules accolées, qui se superposent en spirales. Ces lamelles s'enchevêtrent en bloquant l'écoulement de l'huile. Les additifs s'interposent, soit par adsorption, soit par cocristallisation en favorisant la croissance des lamelles en épaisseur, ce qui les rend moins aptes à l'enchevêtrement. This article begins by describing the main chemical families of pour-point depressant additives. The normal paraffins remaining in the oil after dewaxing crystallize at low temperature in the form of very thin platelets made up of several layers of associated molecules that are superposed in spirals. These platelets become entangled and prevent the flow of oil. Additives work either by adsorption or by cocrystallization, enhancing the growth of lamella thickness, thus making them less capable of becoming entangled.

  16. Hydration layer dynamics and association mechanisms of food and antifreeze proteins : A Molecular Dynamics and Transition Path Sampling study

    NARCIS (Netherlands)

    Brotzakis, Z.F.


    By the time the reader reads this line, billions of protein association events just occurred in our body, such as the ones regulating cell communication, signaling pathways, or in initiating a self-assembly processes, such as tissue fabrication, etc. The timescale of such transitions is slow,

  17. Case Report of a Fatal Antifreeze Ingestion with a Record High Level and Impressive Renal Crystal Deposition

    Directory of Open Access Journals (Sweden)

    Heidi L. Erickson


    Full Text Available Ethylene glycol, methanol, and diethylene glycol are readily available in many household and commercially available products. While these alcohols are relatively nontoxic themselves, their acidic metabolites are toxic and can result in significant morbidity and mortality. Herein we report a lethal case of massive ethylene glycol ingestion in a suicide with a record high level (1254 mg/dL and images of the histologic examination of the kidneys revealing impressive calcium oxalate crystal deposition. Autopsy findings also showed evidence of mild cerebral edema.

  18. Genetic Enhancement of an Anti-Freeze Protein for use as a Substitute for Ethylene Glycol for Aircraft Anti-icing (United States)


    freezing point depression is called a " colligative property", denoting "depending on the collection". 2 ...BACKGROUND: Traditional anti-icing agents are either propylene or ethylene glycol. Glycols are effective in lowering the freezing point of water...mixtures by the phenomenon of freezing point depression based solely on the molal concentration.

  19. Theoretical and experimental study of the antifreeze protein AFP752, trehalose and dimethyl sulfoxide cryoprotection mechanism: correlation with cryopreserved cell viability

    Czech Academy of Sciences Publication Activity Database

    Kratochvílová, Irena; Golan, Martin; Pomeisl, Karel; Richter, Jan; Sedláková, Silvia; Šebera, Jakub; Mičová, Júlia; Falk, Martin; Falková, Iva; Řeha, David; Elliott, J.R.; Varga, K.; Follett, S.E.; Šimek, Daniel


    Roč. 7, č. 1 (2017), s. 352-360 ISSN 2046-2069 R&D Projects: GA ČR(CZ) GA15-05095S; GA MŠk LO1409; GA ČR(CZ) GA14-10279S; GA ČR(CZ) GA16-12454S; GA ČR GBP302/12/G157; GA MZd NV16-29835A Grant - others:FUNBIO(XE) CZ.2.16/3.1.00/21568 Institutional support: RVO:68378271 ; RVO:61388971 ; RVO:68081707 Keywords : X-ray diffraction * DFT * Raman spectroscopy * DSC * cryoprotectants Subject RIV: BO - Biophysics; EE - Microbiology, Virology (MBU-M) OBOR OECD: Biophysics Impact factor: 3.108, year: 2016

  20. [Crystallization of aqueous solutions of cryoprotectors from Atlantic cod and lake Gammarus]. (United States)

    Andreev, A A; Petropavlov, N N


    A comparative analysis of two types of cryoprotectants, antifreeze glycoproteins from Atlantic cod (Gadus morhua) and carbohydrates from hemolymph of amphypoid crustacean (Gammarus lacustris) has been performed. Both glycoprotein and carbohydrate antifreezes effectively decreased the freezing point of water solutions and diminished the size of ice crystals formed. Noncolligative and colligative mechanisms of action are characteristic correspondingly for glycoproteins and carbohydrates.

  1. 75 FR 32754 - Certain New Chemicals; Receipt and Status Information (United States)


    ... Chemical (S) Ultra violet (S) 2-propenoic acid, Industries I, LLC. curable wood 2-methyl-, 2- coatings...; antifreeze in paints and coatings; coalecing aid - inks and coatings P-10-0310 03/30/10 06/27/10 Futurefuel... surfactants solvent; 4-hydroxymethyl-1,3- antifreeze in paints dioxolane and coatings; coalecing aid - inks...

  2. 75 FR 39520 - Certain New Chemicals; Receipt and Status Information (United States)


    ... 06/15/10 09/12/10 Organic Dyestuffs (S) Distribution of (S) Benzenesulfonic Corporation acid dye for... used by industrial customers up to 90%); antifreeze in paints and coatings (to be used by industrial...); solvent (to be used by industrial customers up to 90%); antifreeze in paints and coatings (to be used by...

  3. Presidential Green Chemistry Challenge: 2006 Academic Award (United States)

    Presidential Green Chemistry Challenge 2006 award winner, Professor Galen J. Suppes, developed a process to convert waste glycerin from biodiesel production into propylene glycol to replace ethylene glycol in antifreeze.

  4. Acidosis (United States)

    ... by aspirin, ethylene glycol (found in antifreeze), or methanol Severe dehydration Lactic acidosis is a buildup of ... M. is among the first to achieve this important distinction for online health information and services. Learn ...

  5. Medications: Using Them Safely (United States)

    ... it is given. Never use tableware or a kitchen spoon to measure medication because these don't ... expired medicines. These are the places where antifreeze, oil, and used batteries are taken for proper disposal. ...

  6. Targeted expression of redesigned and codon optimised synthetic gene leads to recrystallisation inhibition and reduced electrolyte leakage in spring wheat at sub-zero temperatures. (United States)

    Khanna, Harjeet K; Daggard, Grant E


    Antifreeze proteins (AFPs) adsorb to ice crystals and inhibit their growth, leading to non-colligative freezing point depression. Crops like spring wheat, that are highly susceptible to frost damage, can potentially be made frost tolerant by expressing AFPs in the cytoplasm and apoplast where ice recrystallisation leads to cellular damage. The protein sequence for HPLC-6 alpha-helical antifreeze protein from winter flounder was rationally redesigned after removing the prosequences in the native protein. Wheat nuclear gene preferred amino acid codons were used to synthesize a recombinant antifreeze gene, rAFPI. Antifreeze protein was targeted to the apoplast using a Murine leader peptide sequence from the mAb24 light chain or retained in the endoplasmic reticulum using C-terminus KDEL sequence. The coding sequences were placed downstream of the rice Actin promoter and Actin-1 intron and upstream of the nopaline synthase terminator in the plant expression vectors. Transgenic wheat lines were generated through micro projectile bombardment of immature embryos of spring wheat cultivar Seri 82. Levels of antifreeze protein in the transgenic lines without any targeting peptide were low (0.06-0.07%). The apoplast-targeted protein reached a level of 1.61% of total soluble protein, 90% of which was present in the apoplast. ER-retained protein accumulated in the cells at levels up to 0.65% of total soluble proteins. Transgenic wheat line T-8 with apoplast-targeted antifreeze protein exhibited the highest levels of antifreeze activity and provided significant freezing protection even at temperatures as low as -7 degrees C.

  7. First report of suspected ethylene glycol poisoning in 2 dogs in South Africa : clinical communication

    Directory of Open Access Journals (Sweden)

    N. Keller


    Full Text Available Ethylene glycol (anti-freeze toxicity is a serious emergency in both veterinary and human medicine. Ethylene glycol (E/G is the active anti-freeze principle in radiator water additives. It is odourless, colourless and has a sweet taste. As little as 5 mℓ or 20 mℓ is sufficient to kill a cat or a dog, respectively. Ethylene glycol is rapidly absorbed and metabolised in the liver to oxalate, which is deposited as calcium oxalate in the kidneys causing irreversible damage. This report describes 2 dogs that were suspected to have ingested ethylene glycol. The report contains a description of the 3 stages of ethylene glycol toxicity as well as a short discussion of the treatment. Public awareness about the dangers of anti-freeze will help in limiting exposure of pets and humans to this potentially fatal toxin. Veterinarians need to be aware of anti-freeze toxicity as delayed recognition and treatment will lead to the death of the patient.

  8. Journal of Chemical Sciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    In this paper, seventeen different fish Antifreeze Proteins (AFPs) retrieved from Swiss-Prot database are analysed and characterized using In silico tools. Primary structure analysis shows that most of the AFPs are hydrophobic in nature due to the high content of non-polar residues. The presence of 11 cysteines in the ...

  9. Green synthesis of tri/tetrasubstituted 1H-imidazoles and 2,3 ...

    Indian Academy of Sciences (India)

    cosmetics, pesticides, disinfectants, agrochemicals, dy- estuffs and antifreeze.1 In particular, derivatives of imi- dazoles and hydroquinazolins are used as ... lar processes.18 These exhibit a wide range of biological activities, such as antitumor, antibiotic, antidefibrillatory, antipyretic, analgesic, antihypertonic, diuretic, antihis ...

  10. Classical Bahavior of Alumina (Al2O3) Nanofluids in Antifrogen N with Experimental Evidence

    NARCIS (Netherlands)

    Saleemi, M.; Vanapalli, Srinivas; Nikkam, N.; Toprak, M.S.; Muhammed, M.


    A nanofluid is a suspension containing nanoparticles in conventional heat transfer fluids. This paper reports on an investigation of alumina (Al2O3) nanoparticles in Antifrogen N, also called AFN, which is a popular antifreeze coolant consisting primarily of ethylene glycol and other additives to

  11. 76 FR 38169 - Toxic Substances Control Act Chemical Testing; Receipt of Test Data (United States)


    ...; solvent for alkyd and phenotic resins; cellulosics; blending agent for lacquers, paints, and varnishes...] - cellulose acetate, Diacetone Alcohol In various oils, resins, Vitro in Human Skin. waxes, fats, dyes, tars... gold leaf, dyeing mixtures, antifreeze mixtures, extraction of resins and waxes, preservative for...

  12. Comparison of arboreal beetle catches in wet and dry collection cups with lindgren multiple funnel traps (United States)

    Daniel R. Miller; Donald A. Duerr


    We compared the effectiveness of a dry collection cup (with an insecticide killing strip) to a wet collection cup (containing antifreeze) for use with Lindgren multiple-funnel traps in catching several common species of bark and wood-boring beetles, and their associates in southern pine forests. All traps were baited with either the binary combination of ethanol and...

  13. Bay-scale population structure in coastal Atlantic cod in Labrador and Newfoundland, Canada

    DEFF Research Database (Denmark)

    Ruzzante, D.E.; Wroblewski, J.S.; Taggart, C.T.


    Polymorphisms at five microsatellite DNA loci provide evidence that Atlantic cod Gadus morhua inhabiting Gilbert Bay, Labrador are genetically distinguishable from offshore cod on the north- east Newfoundland shelf and from inshore cod in Trinity Bay, Newfoundland. Antifreeze activity in the bloo...

  14. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Volume 32 Issue 2 March 2007 pp 251-260 Articles. Antisense expression of a gene encoding a calcium-binding protein in transgenic tobacco leads to altered morphology and enhanced chlorophyll ... 2014 pp 931-944 Reviews. Antifreeze proteins enable plants to survive in freezing conditions · Ravi Gupta Renu Deswal.

  15. 48 CFR 8.1103 - Contract requirements. (United States)


    ... contractor or the Government for furnishing gasoline, motor oil, antifreeze, and similar items. (e) Unless it... PLANNING REQUIRED SOURCES OF SUPPLIES AND SERVICES Leasing of Motor Vehicles 8.1103 Contract requirements. Contracting officers shall include the following items in each contract for leasing motor vehicles: (a) Scope...

  16. Res Dec 07 Final 05-12-07

    Indian Academy of Sciences (India)



    Dec 5, 2007 ... glycerol and aminoacids such as alanine and proline. They are believed to stabilize cellular proteins and membranes at low temperature and hence called cryoprotectants. The cryoprotec- tive role of glycine betaine (GB) in bacteria is beginning to be understood. Antifreeze Proteins: An Overview. Birds ...

  17. Resonance – Journal of Science Education | News

    Indian Academy of Sciences (India)

    Maria Goeppert Mayer · Bikramjit Banerjee · More Details Fulltext PDF. pp 12-24 General Article. Mayer-Jensen Shell Model and Magic Numbers - An Independent Nucleon Model with Spin-Orbit Coupling · R Velusamy · More Details Fulltext PDF. pp 25-30 General Article. Antifreeze Proteins of Bacteria · M K Chattopadhyay.

  18. Fulltext PDF

    Indian Academy of Sciences (India)


    12. Mayer-Jensen Shell Model and Magic Numbers. An Independent Nucleon Model with Spin-Orbit. Coupling. R Velusamy. 46. 25 Antifreeze Proteins of Bacteria. MK Chattopadhyay. Collimator. 31 Seven Different Proofs of the Irrationality of 12. VG Tikekar. Prism Table. Telescope. 40 Getting Sloshed in Outer Space.

  19. Mapping of QTLs for frost tolerance and heading time using SSR ...

    African Journals Online (AJOL)



    Oct 19, 2008 ... Heredity 42: 125-132. Chmielewicz KM, Manly KF (2002). Software for genetic mapping for mendelian markers and quantitative trait loci. Roswell Park Cancer. Institute. Chun JU, Yu XM, Griffith M (1998). Genetic studies of antifreeze proteins and their correlation with winter survival in wheat. Euphytica.

  20. Cloning and mRNA expression pattern analysis under low ...

    African Journals Online (AJOL)



    Jul 13, 2011 ... mesophyll, epidermal cells and phloem parenchyma, and are accumulated indirectly as part of growth and development (Yeh et al., 2000). However, reports about anti-freeze protein evolution of EAPP chitinase of. Dongmu-70 rye and expression pattern analysis under low temperature stress are few.

  1. Data from thermal testing of the Open Source Cryostage

    DEFF Research Database (Denmark)

    Buch, Johannes Lørup; Ramløv, Hans


    The data presented here is related to the research article "An open source cryostage and software analysis method for detection of antifreeze activity" (Buch and Ramløv, 2016) [1]. The design of the Open Source Cryostage (OSC) is tested in terms of thermal limits, thermal efficiency and electrical...

  2. 21 CFR 522.1004 - Fomepizole. (United States)


    ... Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS...) Conditions of use in dogs—(1) Amount. 20 milligrams per kilogram (mg/kg) of body weight intravenously... antidote for ethylene glycol (antifreeze) poisoning in dogs who have ingested or are suspected of having...

  3. 13 Animal Emergencies That Should Receive Immediate Veterinary Consultation and/or Care (United States)

    ... has eaten something poisonous (such as antifreeze, xylitol, chocolate, rodent poison, etc.) Seizures and/or staggering Fractured bones, severe lameness or inability to move leg(s) Obvious signs of pain or extreme anxiety Heat stress or heatstroke Severe vomiting or diarrhea – more than ...

  4. Ti/TiO2 nanotube array electrode as a new sensor to ...

    Indian Academy of Sciences (India)

    EG (EG oxidation current in the UV irradiation) was assessed using the hydrodynamic photoamperometric method in the phosphate buffers. ... antifreeze, engine coolants, dyes, some types of inks and other chemicals. Given the ... treated Ti sheet was served as the anode and a Pt electrode was served as the cathode, and ...

  5. Attraction of Bactrocera dorsalis (Diptera: Tephritidae) and nontarget insects to methyl eugenol bucket traps with different preservative fluids on Oahu Island, Hawaiian Islands. (United States)

    Uchida, Grant K; Mackey, Bruce E; McInnis, Donald O; Vargas, Roger I


    Attraction of oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), and nontarget insects to preservative fluids ethylene glycol antifreeze, propylene glycol antifreeze, or mineral oil in bucket traps that contained captured decaying male oriental fruit flies, a male lure (methyl eugenol), and a toxicant (DDVP vapor insecticidal strip) were compared with dry control traps. Significantly (P < 0.05) greater numbers of B. dorsalis were captured in propylene glycol antifreeze traps than in other attractant trap types. Among attractant trap types with lowest negative impacts on nontarget insects, control traps captured significantly lower numbers of three species and one morphospecies of scavenger flies, one species of plant-feeding fly, and one species each of sweet-and lipid-feeding ants. Mineral oil traps captured significantly lower numbers of two species of scavengers flies and one morphospecies of plant-feeding fly, and one species of sweet-feeding ant. Because of the fragile nature of endemic Hawaiian insect fauna, the propylene glycol antifreeze bucket trap is best suited for use in environments (e.g., non-native habitats) where endemic species are known to be absent and mineral oil traps are more suited for minimizing insect captures in environmentally sensitive habitats.

  6. 49 CFR 178.810 - Drop test. (United States)


    ... liquids must be kept in the liquid state, if necessary, by the addition of anti-freeze. Water/anti-freeze solutions with a minimum specific gravity of 0.95 for testing at −18 °C (0 °F) or lower are considered... material having essentially the same physical characteristics. (3) The specific gravity and viscosity of a...

  7. A Hybrid Computational-Experimental Framework for Microbial Chemical Synthesis via Enzyme Channeling (United States)


    in the market size annually. Some typical uses of 1,2-PD are in unsaturated polyester resins, functional fluids (antifreeze, de-icing, and heat...its natural ligand, the freely diffusible quorum signaling molecule 3-oxooctanyl-l-homoserine lactone (OHHL), the TraR protein is a monomer that is

  8. Necrosis and haemorrhage of the putamen in methanol poisoning shown on MRI

    International Nuclear Information System (INIS)

    Kuteifan, K.; Gutbub, A.M.; Laplatte, G.; Oesterle, H.; Tajahmady, T.


    Methanol, a highly toxic substance, is used as an industrial solvent and in automobile antifreeze. Acute methanol poisoning produces severe metabolic acidosis and serious neurologic sequelae. We describe a 50-year-old woman with accidental methanol intoxication who was in a vegetative state. MRI showed haemorrhagic necrosis of the putamina and oedema in the deep white matter. (orig.)

  9. Green synthesis of tri/tetrasubstituted 1H-imidazoles and 2,3 ...

    Indian Academy of Sciences (India)

    cosmetics, pesticides, disinfectants, agrochemicals, dy- estuffs and antifreeze.1 In particular, derivatives of imi- dazoles and hydroquinazolins are used as reagents in industry.2 The five-membered imidazole ring is one of the important substructures found in a large number of natural products and pharmacologically active ...

  10. Inhibition of ice recrystallization and cryoprotective activity of wheat proteins in liver and pancreatic cells. (United States)

    Chow-Shi-Yée, Mélanie; Briard, Jennie G; Grondin, Mélanie; Averill-Bates, Diana A; Ben, Robert N; Ouellet, François


    Efficient cryopreservation of cells at ultralow temperatures requires the use of substances that help maintain viability and metabolic functions post-thaw. We are developing new technology where plant proteins are used to substitute the commonly-used, but relatively toxic chemical dimethyl sulfoxide. Recombinant forms of four structurally diverse wheat proteins, TaIRI-2 (ice recrystallization inhibition), TaBAS1 (2-Cys peroxiredoxin), WCS120 (dehydrin), and TaENO (enolase) can efficiently cryopreserve hepatocytes and insulin-secreting INS832/13 cells. This study shows that TaIRI-2 and TaENO are internalized during the freeze-thaw process, while TaBAS1 and WCS120 remain at the extracellular level. Possible antifreeze activity of the four proteins was assessed. The "splat cooling" method for quantifying ice recrystallization inhibition activity (a property that characterizes antifreeze proteins) revealed that TaIRI-2 and TaENO are more potent than TaBAS1 and WCS120. Because of their ability to inhibit ice recrystallization, the wheat recombinant proteins TaIRI-2 and TaENO are promising candidates and could prove useful to improve cryopreservation protocols for hepatocytes and insulin-secreting cells, and possibly other cell types. TaENO does not have typical ice-binding domains, and the TargetFreeze tool did not predict an antifreeze capacity, suggesting the existence of nontypical antifreeze domains. The fact that TaBAS1 is an efficient cryoprotectant but does not show antifreeze activity indicates a different mechanism of action. The cryoprotective properties conferred by WCS120 depend on biochemical properties that remain to be determined. Overall, our results show that the proteins' efficiencies vary between cell types, and confirm that a combination of different protection mechanisms is needed to successfully cryopreserve mammalian cells. © 2016 The Protein Society.

  11. Bioinspiration From Nano to Micro Scales

    CERN Document Server


    Methods in bioinspiration and biomimicking have been around for a long time. However, due to current advances in modern physical, biological sciences, and technologies, our understanding of the methods have evolved to a new level. This is due not only to the identification of mysterious and fascinating phenomena but also to the understandings of the correlation between the structural factors and the performance based on the latest theoretical, modeling, and experimental technologies. Bioinspiration: From Nano to Micro Scale provides readers with a broad view of the frontiers of research in the area of bioinspiration from the nano to macroscopic scales, particularly in the areas of biomineralization, antifreeze protein, and antifreeze effect. It also covers such methods as the lotus effect and superhydrophobicity, structural colors in animal kingdom and beyond, as well as behavior in ion channels. A number of international experts in related fields have contributed to this book, which offers a comprehensive an...

  12. Decoration of Chondroitin Polysaccharide with Threonine: Synthesis, Conformational Study, and Ice-Recrystallization Inhibition Activity. (United States)

    Laezza, Antonio; Casillo, Angela; Cosconati, Sandro; Biggs, Caroline I; Fabozzi, Antonio; Paduano, Luigi; Iadonisi, Alfonso; Novellino, Ettore; Gibson, Matthew I; Randazzo, Antonio; Corsaro, Maria M; Bedini, Emiliano


    Several threonine (Thr)- and alanine (Ala)-rich antifreeze glycoproteins (AFGPs) and polysaccharides act in nature as ice recrystallization inhibitors. Among them, the Thr-decorated capsular polysaccharide (CPS) from the cold-adapted Colwellia psychrerythraea 34H bacterium was recently investigated for its cryoprotectant activity. A semisynthetic mimic thereof was here prepared from microbial sourced chondroitin through a four-step strategy, involving a partial protection of the chondroitin polysaccharide as a key step for gaining an unprecedented quantitative amidation of its glucuronic acid units. In-depth NMR and computational analysis suggested a fairly linear conformation for the semisynthetic polysaccharide, for which the antifreeze activity by a quantitative ice recrystallization inhibition assay was measured. We compared the structure-activity relationships for the Thr-derivatized chondroitin and the natural Thr-decorated CPS from C. psychrerythraea.

  13. Hemodiafiltration efficacy in treatment of methanol and ethylene glycol poisoning in a 2-year-old girl. (United States)

    Szmigielska, Agnieszka; Szymanik-Grzelak, Hanna; Kuźma-Mroczkowska, Elżbieta; Roszkowska-Blaim, Maria


    Every year about 2.4 million people in USA are exposed to toxic substances. Many of them are children below 6 years of age. Majority of poisonings in children are incidental and related to household products including for example drugs, cleaning products or antifreeze products. Antifreeze solutions contain ethylene glycol and methanol. Treatment of these toxic substances involves ethanol administration, fomepizole, hemodialysis and correction of metabolic acidosis. The aim of the study was to check the efficacy of continuous venovenous hemodiagiltration in intoxication with ethylene glycol and methanol. One year and 7 months old girl after intoxication with ethylene glycol and methanol was treated with continuous venovenous hemodiafiltration instead of hemodialysis because of technical problems (circulatory instability). Intravenous ethanol infusion with hemodialtration resulted in rapid elimination of methanol from the body and significantly reduced blood ethylene glycol level. Continuous venovenous hemodiafiltration can be helpful in treatment of ethylene glycol and methanol intoxication.

  14. An Overview of Conceptual Frameworks (United States)


    the water will freeze , add antifreeze. NSM KNOW (v) KNOWS, KNEW, KNOWN To be sure of data, to have data “ready” to use If you know the clearance...illustrate these ideas. The Tropes of a ripening tomato are its color, its mass, its shape, etc. The change of a ripening tomato just pertains to...different Thick Objects representing the tomato and its Tropes. That means the Thick Objects are wholes centered around the bundle of core characteristics

  15. Aglomerularism in Antarctic fish. (United States)

    Dobbs, G H; Lin, Y; DeVries, A L


    Urine formation in antarctic bony fish does not involve glomerular filtration. Evidence for aglomerularism came from both direct observation of kidney serial sections by light microscopy and the low concentrations of inulin labeled with carbon-14 that were excreted into the urine when this renal clearance tracer was injected into the bloodstream via a cannula implanted in the caudal vein. Aglomerularism most likely prevents urinary loss of glycoproteins with biological antifreeze properties.

  16. Emission of ammonia from indoor concrete wall and assessment of human exposure. (United States)

    Bai, Z; Dong, Y; Wang, Z; Zhu, T


    Addition of urea-based antifreeze admixtures during cement mixing can make it possible to produce concrete cement in construction of buildings in cold weather; this, however, has led to increasing indoor air pollution due to continuous transformation and emission from urea to gaseous ammonia in indoor concrete wall. It is believed that ammonia is harmful to human body and exposure to ammonia can cause some serious symptoms such as headaches, burns, and even permanent damage to the eyes and lungs. In order to understand the emission of ammonia from indoor concrete wall in civil building and assess the health risk of people living in these buildings, the experimental pieces of concrete wall were first prepared by concreting cement and urea-based antifreeze admixtures to simulate the indoor wall in civil building in this work. Then environmental chamber was adopted for studying the effect of temperature, relative humility and air exchange rate on emission of ammonia from experimental pieces of concrete wall. Also the field experiment was made at selected rooms in given civil buildings. Exposure and potential dose of adult and children exposed to indoor/outdoor ammonia in summer and in winter are calculated and evaluated by using Scenario Evaluation Approach. The results indicated that high air exchange rate leads to decreased ammonia concentration, and elevation of temperature causes increasing ammonia concentration and volatilizing rate in chamber. The complete emission of ammonia from the wall containing urea-based antifreeze admixtures needs more than 10 years in general. Ventilating or improving air exchange can play a significant role in reducing ammonia concentration in actual rooms in field experiments. Urea-based antifreeze admixtures in concrete wall can give rise to high exposure and potential dose, especially in summer. Generally, adults have a high potential dose than children, while children have personal average dose rate beyond adults in the same

  17. Changing climate and microbial resources in polar realms

    Digital Repository Service at National Institute of Oceanography (India)

    LokaBharathi, P.A.

    relationship that exists between oceanic primayy productivity and sea surface OMS concentrations and justifies more intense observations on a dynamcc scale . Besides contributing to climate modulating processe,, microbial metabolites could contribute to a... | 49 SaGAA 2011 Antifreeze proteins have been used from microbial sources and higher organisms for processing low fat food items like ice cream. Such proteins can also be used to improve surgery and tissue transplants. Plants could also be engineered...

  18. Ethylene glycol poisoning

    African Journals Online (AJOL)

    Ethylene glycol poisoning. A 22-year-old male presented to the emergency centre after drinking 300 ml of antifreeze. Clinical examination was unremarkable except for a respiratory rate of 28 bpm, GCS of 9 and slight nystagmus. Arterial blood gas revealed: pH 7.167, pCO2. 3.01 kPa, pO2 13.0 kPa (on room air), HCO3-.

  19. Facing Stormwater Management Challenges at a Southeastern Army Installation: US Army Garrison Fort Gordon (United States)


    the Solution to Stormwater Pollution at Fort Gordon ~trriGffiG 01 ~~~ • Oil , grease , heavy metals and toxic chemicals from motor vehicles...etc.) into the storm drains. Sinw stormwater drains are not oonnectacl to tl’e wastewater treatment system, the polluted stormwat:er Is trcn...examples of illicit discharge materials include paint, oil , antifreeze, yancl waste, etc. These pollutants degrade water qufllity anrl threaten

  20. Industrial Processes to Reduce Generation of Hazardous Waste at DoD Facilities. Phase III Report. Appendix C. Workshop Manual Centralized Vehicle Wash Racks and Scheduled Maintenance Facilities, Fort Lewis, Washington. (United States)


    CLEANING ENGINE COMPARTMENTS, CHANGING OIL AND ANTIFREEZE, CHASSIS LUBRICATION, ETC., WITH THE ELIMINATION OF ENVIRONMENTAL POLLUTION COMMONLY...such as Fork Lifts or Wheel Loading V. and Other Critical Loads on Floor Slabs: Heavy abrasive use from X60 Tank and other track vechile . (13) Fallout...Is the product of engine and chassis cleaning operations using high pressure hot water washers. These washers DO NOT use detergents or chemical

  1. Molecular Characterization, Structural Modeling, and Evaluation of Antimicrobial Activity of Basrai Thaumatin-Like Protein against Fungal Infection. (United States)

    Yasmin, Nusrat; Saleem, Mahjabeen; Naz, Mamoona; Gul, Roquyya; Rehman, Hafiz Muzzammel


    A thaumatin-like protein gene from Basrai banana was cloned and expressed in Escherichia coli . Amplified gene product was cloned into pTZ57R/T vector and subcloned into expression vector pET22b(+) and resulting pET22b-basrai TLP construct was introduced into E. coli BL21. Maximum protein expression was obtained at 0.7 mM IPTG concentration after 6 hours at 37°C. Western blot analysis showed the presence of approximately 20 kDa protein in induced cells. Basrai antifungal TLP was tried as pharmacological agent against fungal disease. Independently Basrai antifungal protein and amphotericin B exhibited their antifungal activity against A. fumigatus ; however combined effect of both agents maximized activity against the pathogen. Docking studies were performed to evaluate the antimicrobial potential of TLP against A. fumigatus by probing binding pattern of antifungal protein with plasma membrane ergosterol of targeted fungal strain. Ice crystallization primarily damages frozen food items; however addition of antifreeze proteins limits the growth of ice crystal in frozen foods. The potential of Basrai TLP protein, as an antifreezing agent, in controlling the ice crystal formation in frozen yogurt was also studied. The scope of this study ranges from cost effective production of pharmaceutics to antifreezing and food preserving agent as well as other real life applications.

  2. Molecular Characterization, Structural Modeling, and Evaluation of Antimicrobial Activity of Basrai Thaumatin-Like Protein against Fungal Infection

    Directory of Open Access Journals (Sweden)

    Nusrat Yasmin


    Full Text Available A thaumatin-like protein gene from Basrai banana was cloned and expressed in Escherichia coli. Amplified gene product was cloned into pTZ57R/T vector and subcloned into expression vector pET22b(+ and resulting pET22b-basrai TLP construct was introduced into E. coli BL21. Maximum protein expression was obtained at 0.7 mM IPTG concentration after 6 hours at 37°C. Western blot analysis showed the presence of approximately 20 kDa protein in induced cells. Basrai antifungal TLP was tried as pharmacological agent against fungal disease. Independently Basrai antifungal protein and amphotericin B exhibited their antifungal activity against A. fumigatus; however combined effect of both agents maximized activity against the pathogen. Docking studies were performed to evaluate the antimicrobial potential of TLP against A. fumigatus by probing binding pattern of antifungal protein with plasma membrane ergosterol of targeted fungal strain. Ice crystallization primarily damages frozen food items; however addition of antifreeze proteins limits the growth of ice crystal in frozen foods. The potential of Basrai TLP protein, as an antifreezing agent, in controlling the ice crystal formation in frozen yogurt was also studied. The scope of this study ranges from cost effective production of pharmaceutics to antifreezing and food preserving agent as well as other real life applications.

  3. Optimization Of Freeze-Dried Starter For Yogurt By Full Factorial Experimental Design

    Directory of Open Access Journals (Sweden)

    Chen He


    Full Text Available With the rapidly development of fermented milk product, it is significant for enhancing the performance of starter culture. This paper not only investigated the influence of anti-freeze factors and freeze-drying protective agents on viable count, freeze-drying survival rate and yield of Lactobacillus bulgaricus (LB and Streptococcus thermophilus (ST, but also optimized the bacteria proportion of freeze-dried starter culture for yogurt by full factorial experimental design. The results showed as following: the freeze-drying protective agents or anti-freeze factors could enhanced survival rate of LB and ST; the freeze-dried LB and ST powders containing both of anti-freeze factors and freeze-drying protective agents had higher viable count and freeze-drying survival rate that were 84.7% and 79.7% respectively; In terms of fermentation performance, the best group of freeze-dried starter for yogurt was the compound of LB3 and ST2.



    ÖZSOLAK, Onur; ESEN, Mehmet


    In the following study, 12 m2 test chamber was heated by solar and ground source heat pump under the physical conditions of Elazığ. In order to place slinky heat exchanger pipes, a hole was dug with 1 meter width, 2 meters depth and 15 meters length. Slinky pipes were put horizontally in the hole and water-antifreeze mixture was circulated with the circulating pump in the slinky heat exchanger. The heat taken from the ground was transferred into the environment to be heated through the heat p...

  5. Data from thermal testing of the Open Source Cryostage

    DEFF Research Database (Denmark)

    Buch, Johannes Lørup; Ramløv, Hans


    The data presented here is related to the research article “An open source cryostage and software analysis method for detection of antifreeze activity” (Buch and Ramløv, 2016) [1]. The design of the Open Source Cryostage (OSC) is tested in terms of thermal limits, thermal efficiency and electrical...... efficiency. This article furthermore includes an overview of the electrical circuitry and a flowchart of the software program controlling the temperature of the OSC. The thermal efficiency data is presented here as degrees per volt and maximum cooling capacity....

  6. Data from thermal testing of the Open Source Cryostage

    Directory of Open Access Journals (Sweden)

    Johannes Lørup Buch


    Full Text Available The data presented here is related to the research article “An open source cryostage and software analysis method for detection of antifreeze activity” (Buch and Ramløv, 2016 [1]. The design of the Open Source Cryostage (OSC is tested in terms of thermal limits, thermal efficiency and electrical efficiency. This article furthermore includes an overview of the electrical circuitry and a flowchart of the software program controlling the temperature of the OSC. The thermal efficiency data is presented here as degrees per volt and maximum cooling capacity.

  7. Storage products and tissue interaction in the ovule of Pinus silvestris (L.

    Directory of Open Access Journals (Sweden)

    F. M. Engels


    Full Text Available The organel-sequence in ovular cells of Pinus silvestris was investigated by light- and electronmicroscopy during the post-pollination and pre-fertilization period. Changes in starch and lipid storage suppose starch to be a pool for lipid synthesis and a reserve for ovule development. The base nucellus plays an important role in the distribution of metabolites all over the ovular tissues. Lipid, starch and callose are of interest for the cells to protect them against low temperatures by means of isolation, antifreeze and plug formation respectively.

  8. TA-60-1 Heavy Equipment Shop Areas SWPPP Rev 2 Jan 2017-Final

    Energy Technology Data Exchange (ETDEWEB)

    Burgin, Jillian Elizabeth [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)


    The primary activities and equipment areas at the facility that are potential stormwater pollution sources include; The storage of vehicles and heavy equipment awaiting repair; or repaired vehicles waiting to be picked up; The storage and handling of oils, anti-freeze, solvents, degreasers, batteries and other chemicals for the maintenance of vehicles and heavy equipment; and Equipment cleaning operations including exterior vehicle wash-down. Steam cleaning is only done on the steam cleaning pad area located at the north east end of Building 60-0001.

  9. Unusual Clinical Presentation of Ethylene Glycol Poisoning: Unilateral Facial Nerve Paralysis

    Directory of Open Access Journals (Sweden)

    Eray Eroglu


    Full Text Available Ethylene glycol (EG may be consumed accidentally or intentionally, usually in the form of antifreeze products or as an ethanol substitute. EG is metabolized to toxic metabolites. These metabolites cause metabolic acidosis with increased anion gap, renal failure, oxaluria, damage to the central nervous system and cranial nerves, and cardiovascular instability. Early initiation of treatment can reduce the mortality and morbidity but different clinical presentations can cause delayed diagnosis and poor prognosis. Herein, we report a case with the atypical presentation of facial paralysis, hematuria, and kidney failure due to EG poisoning which progressed to end stage renal failure and permanent right peripheral facial nerve palsy.

  10. Probability of freezing in the freeze-avoiding beetle larvae Cucujus clavipes puniceus (Coleoptera: Cucujidae) from interior Alaska. (United States)

    Sformo, T; McIntyre, J; Walters, K R; Barnes, B M; Duman, J


    Freeze-avoiding insects must resist freezing or die. A suite of adaptations to low temperatures, including the production of antifreeze proteins, colligative antifreezes (polyols), and dehydration allows most individuals to prevent freezing below the lowest ambient temperatures experienced in situ; however, there can be a wide variance in the minimum temperatures that individuals of freeze-avoiding species reach before freezing. We used logistic regression to explore factors that affect this variance and to estimate the probability of freezing in larvae of the freeze-avoiding beetle Cucujus clavipes puniceus. We hypothesized that water content ≤0.5 mg mg(-1) dry mass would lead to deep supercooling (avoidance of freezing below -58°C). We found a significant interaction between water content and ambient below-snow temperature and a significant difference between individuals collected from two locations in Alaska: Wiseman and Fairbanks. Individuals collected in Wiseman deep supercooled with greater water content and to a greater range of ambient temperatures than individuals collected in Fairbanks, leading to significantly different lethal water contents associated with 50% probability of freezing. Copyright © 2011 Elsevier Ltd. All rights reserved.

  11. Surrogate alcohol containing methanol, social deprivation and public health in Novosibirsk, Russia. (United States)

    Neufeld, Maria; Lachenmeier, Dirk; Hausler, Thomas; Rehm, Jürgen


    Surrogate alcohol, i.e. alcohol not intended or not officially intended for human consumption, continues to play an important role in alcohol consumption in Russia, especially for people with alcohol dependence. Among the different types of surrogate alcohol, there are windshield washer antifreeze liquids; these products are the cheapest kinds of non-beverage alcohol available and thus likely to be used by the most deprived and marginalised groups such as homeless people with alcohol dependence. Although it is well known, that non-beverage alcohol is used for consumption by various groups in Russia, and although there are laws to prohibit the use of methanol as part of windshield washer antifreeze liquids for the very reason that such products could be used as surrogate alcohol, we detected products in retail sale which were a mix of water and methanol only. Methanol poses serious health threats including blindness and death, and there had been repeated methanol deaths from surrogate alcohol in Russia over the last years. If law-enforcement does not change for surrogate products, we can expect more methanol-resulting deaths in the most deprived and marginalized groups of people with alcohol dependence in Russia. In addition, ingredients with questionable safety profiles such as formic acid should also be prohibited in non-beverage alcohol products that are likely to be consumed as surrogate alcohol. Copyright © 2016. Published by Elsevier B.V.

  12. Capture of Anastrepha species (Diptera: Tephritidae) with multilure traps and biolure attractants in Guatemala

    International Nuclear Information System (INIS)

    Martinez, A.J.; Salinas, E.J.; Rendon, P.


    Two trapping systems were compared in a study in Guatemala during the wet season, May through Dec 2001. Trap/lure combinations consisting of green or yellow-based plastic McPhail-like traps baited with a synthetic 2-component lure (putrescine and ammonium acetate) and 300 mL of propylene glycol antifreeze as a preservative were compared to the traditional glass McPhail baited with torula yeast/borax and 300 mL of water. Both systems captured several key Anastrepha species including Anastrepha ludens Loew, A. obliqua, Macquart, A. serpentina Weidemann, A. striata Schiner, A. distincta Greene, A. fraterculus Weidemann as well as Ceratitis capitata Weidemann. Additionally, 13 other Anastrepha spp. were captured with the synthetic lure. The plastic traps captured more key flies than the McPhail trap except for A. distincta where there were no significant differences between the yellow-based plastic trap and the McPhail trap and no significant differences between any trap and lure for trapping A. fraterculus. The synthetic lure lasted 10 weeks. The sex ratio was female-biased for almost all captured key species in both systems. Moreover, there were significant numbers of captured nontarget insects in all traps; however, the captured flies in those traps with the synthetic lure were not adversely affected by these insects. Propylene glycol-based antifreeze was a superior preservative when compared to borax/water. (author) [es

  13. The genetic characteristics in cytology and plant physiology of two wheat (Triticum aestivum) near isogenic lines with different freezing tolerances. (United States)

    Wang, Wenqiang; Hao, Qunqun; Wang, Wenlong; Li, Qinxue; Wang, Wei


    Freezing tolerance in taft plants relied more upon an ABA-independent- than an ABA-dependent antifreeze signaling pathway. Two wheat (Triticum aestivum) near isogenic lines (NIL) named tafs (freezing sensitivity) and taft (freezing tolerance) were isolated in the laboratory and their various cytological and physiological characteristics under freezing conditions were studied. Proplastid, cell membrane, and mitochondrial ultrastructure were less damaged by freezing treatment in taft than tafs plants. Chlorophyll, ATP, and thylakoid membrane protein contents were significantly higher, but malondialdehyde content was significantly lower in taft than tafs plants under freezing condition. Antioxidant capacity, as indicated by reactive oxygen species accumulation and antioxidant enzyme activity, and the relative gene expression were significantly greater in taft than tafs plants. Soluble sugars and abscisic acid (ABA) contents were significantly higher in taft plants than in tafs plants under both normal and freezing conditions. The upregulated expression levels of certain freezing tolerance-related genes were greater in taft than tafs plants under freezing treatment. The addition of sodium tungstate, an ABA synthesis inhibitor, led to only partial freezing tolerance inhibition in taft plants and the down-regulated expression of some ABA-dependent genes. Thus, both ABA-dependent and ABA-independent signaling pathways are involved in the freezing tolerance of taft plants. At the same time, freezing tolerance in taft plants relied more upon an ABA-independent- than an ABA-dependent antifreeze signaling pathway.

  14. Bacterial lipoteichoic acid enhances cryosurvival. (United States)

    Rice, Charles V; Middaugh, Amy; Wickham, Jason R; Friedline, Anthony; Thomas, Kieth J; Scull, Erin; Johnson, Karen; Zachariah, Malcolm; Garimella, Ravindranth


    Antifreeze proteins in fish, plants, and insects provide protection to a few degrees below freezing. Microbes have been found to survive at even lower temperatures, and with a few exceptions, antifreeze proteins are missing. We show that lipoteichoic acid (LTA), a biopolymer in the cell wall of Gram-positive bacteria, can be added to B. subtilis cultures and increase freeze tolerance. At 1 % w/v, LTA enables a 50 % survival rate, similar to the results obtained with 1 % w/v glycerol as measured with the resazurin cell viability assay. In the absence of added LTA or glycerol, a very small number of B. subtilis cells survive freezing. This suggests that an innate freeze tolerance mechanism exists. While cryoprotection can be provided by extracellular polymeric substances, our data demonstrate a role for LTA in cryoprotection. Currently, the exact mode of action for LTA cryoprotection is unknown. With a molecular weight of 3-5 kDa, it is unlikely to enter the cell cytoplasm. However, low temperature microscopy data show small ice crystals aligned along channels of liquid water. Our observations suggest that teichoic acids could protect liquid water within biofilms and planktonic bacteria, augmenting the role of brine while also raising the possibility for survival without brine present.

  15. High Tensile Strength of Engineered β-Solenoid Fibrils via Sonication and Pulling. (United States)

    Peng, Zeyu; Parker, Amanda S; Peralta, Maria D R; Ravikumar, Krishnakumar M; Cox, Daniel L; Toney, Michael D


    We present estimates of ultimate tensile strength (UTS) for two engineered β-solenoid protein mutant fibril structures (spruce budworm and Rhagium inquisitor antifreeze proteins) derived from sonication-based measurements and from force pulling molecular dynamics simulations, both in water. Sonication experiments generate limiting scissioned fibrils with a well-defined length-to-width correlation for the mutant spruce budworm protein and the resultant UTS estimate is 0.66 ± 0.08 GPa. For fibrils formed from engineered R. inquisitor antifreeze protein, depending upon geometry, we estimate UTSs of 3.5 ± 3.2-5.5 ± 5.1 GPa for proteins with interfacial disulfide bonds, and 1.6 ± 1.5-2.5 ± 2.3 GPa for the reduced form. The large error bars for the R. inquisitor structures are intrinsic to the broad distribution of limiting scission lengths. Simulations provide pulling velocity-dependent UTSs increasing from 0.2 to 1 GPa in the available speed range, and 1.5 GPa extrapolated to the speeds expected in the sonication experiments. Simulations yield low-velocity values for the Young's modulus of 6.0 GPa. Without protein optimization, these mechanical parameters are similar to those of spider silk and Kevlar, but in contrast to spider silk, these proteins have a precisely known sequence-structure relationship. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  16. Short communication. Impact of the amino acid proline on the cold hardiness of honey bee, Apis mellifera L.

    Directory of Open Access Journals (Sweden)

    M. Mollaei


    Full Text Available Like many insects, honey bee can increase its cold tolerance through freeze avoidance, using antifreeze proteins (AFPs to lower its supercooling point (SCP. Proline is the most dominant amino acid in honey bee hemolymph, which can be obtained by the insect through feeding. In the current study the antifreeze activity of this amino acid was evaluated on worker honey bees, immediately before the start of cold season. The experiment was established on four treatments including three different concentrations of proline (1%, 3% and 4.35% diluted in 1:1 water sucrose syrup, and the syrup without proline (control. Newly emerged worker honey bees were fed on the mentioned diets for 2 weeks, under cage condition, and then 20 bees from each treatment (cage were selected randomly for determination of cold hardiness inside a cooling bath. Using a CHY data logger, equipped with a K100 sensor attached to the bee’s gaster, the SCP, the amount of released heat and the rate of this release as measures of insect cold hardiness were recorded. Proline significantly reduced honey bees’ SCP. The lowest point, -7.67±0.2646°C, was observed in the concentration of 1% proline. The amount of released heat and the rate of this release were not significantly different across the treatments.

  17. Ethylene Glycol, Hazardous Substance in the Household

    Directory of Open Access Journals (Sweden)

    Jiří Patočka


    Full Text Available Ethylene glycol is a colorless, odorless, sweet-tasting but poisonous type of alcohol found in many household products. The major use of ethylene glycol is as an antifreeze in, for example, automobiles, in air conditioning systems, in de-icing fluid for windshields, and else. People sometimes drink ethylene glycol mistakenly or on purpose as a substitute for alcohol. Ethylene glycol is toxic, and its drinking should be considered a medical emergency. The major danger from ethylene glycol is following ingestion. Due to its sweet taste, peoples and occasionally animals will sometimes consume large quantities of it if given access to antifreeze. While ethylene glycol itself has a relatively low degree of toxicity, its metabolites are responsible for extensive cellular damage to various tissues, especially the kidneys. This injury is caused by the metabolites, glycolic and oxalic acid and their respective salts, through crystal formation and possibly other mechanisms. Toxic metabolites of ethylene glycol can damage the brain, liver, kidneys, and lungs. The poisoning causes disturbances in the metabolism pathways, including metabolic acidosis. The disturbances may be severe enough to cause profound shock, organ failure, and death. Ethylene glycol is a common poisoning requiring antidotal treatment.

  18. Molecular Dynamics at the Interface between Ice and Poly(vinyl alcohol) and Ice Recrystallization Inhibition. (United States)

    Weng, Lindong; Stott, Shannon L; Toner, Mehmet


    Ice formation is a ubiquitous process that poses serious challenges for many areas. Nature has evolved a variety of different mechanisms to regulate ice formation. For example, many cold-adapted species produce antifreeze proteins (AFPs) and/or antifreeze glycoproteins (AFGPs) to inhibit ice recrystallization. Although several synthetic substitutes for AF(G)Ps have been developed, the fundamental principles of designing AF(G)P mimics are still missing. In this study, we explored the molecular dynamics of ice recrystallization inhibition (IRI) by poly(vinyl alcohol) (PVA), a well-recognized ice recrystallization inhibitor, to shed light on the otherwise hidden ice-binding mechanisms of chain polymers. Our molecular dynamics simulations revealed a stereoscopic, geometrical match between the hydroxyl groups of PVA and the water molecules of ice, and provided microscopic evidence of the adsorption of PVA to both the basal and prism faces of ice and the incorporation of short-chain PVA into the ice lattice. The length of PVA, i.e., the number of hydroxyl groups, seems to be a key factor dictating the performance of IRI, as the PVA molecule must be large enough to prevent the joining together of adjacent curvatures in the ice front. The findings in this study will help pave the path for addressing a pressing challenge in designing synthetic ice recrystallization inhibitors rationally, by enriching our mechanistic understanding of IRI process by macromolecules.

  19. Understanding Poly(vinyl alcohol)-Mediated Ice Recrystallization Inhibition through Ice Adsorption Measurement and pH Effects. (United States)

    Burkey, Aaron A; Riley, Christopher L; Wang, Lyndsey K; Hatridge, Taylor A; Lynd, Nathaniel A


    The development of improved cryopreservative materials is necessary to enable complete recovery of living cells and tissue after frozen storage. Remarkably, poly(vinyl alcohol) (PVA) displays some of the same cryoprotective properties as many antifreeze proteins found in cold tolerant organisms. In particular, PVA is very effective at halting the Ostwald ripening of ice, a process that mechanically damages cells and tissue. Despite the large practical importance of such a property, the mechanism by which PVA interacts with ice is poorly understood, hindering the development of improved cryoprotective materials. Herein, we quantitatively evaluated ice growth kinetics in the presence of PVA at different pH conditions and in the presence of a range of neutral salts. We demonstrated that pH, but not salt identity, alters the ability of PVA to halt ice grain coarsening. These observations are consistent with hydrogen-bonding playing a crucial role in PVA-mediated ice recrystallization inhibition. The evolution of the size distribution of ice crystals with annealing was consistent with incomplete surface coverage of ice with PVA. Binding assay measurements of dissolved fluorescently labeled PVA in an ice slurry showed that PVA interacts with ice through weak adsorption (<9%) to the ice crystal surface, which stands in contrast to fluorescently tagged type III antifreeze peptide, which binds strongly (ca. 64%) under the same conditions.


    Directory of Open Access Journals (Sweden)

    V. Cherepnin


    Full Text Available Purpose. The purpose of these studies was to evaluate the effect of the cryoselection of carp sperm of multiple age groups depending on the quality of thawed sperm after modification of the composite cryoprotective medium, which was used for the dilution of native sperm before freezing. Methodology. Coenzyme B12 (cobamamide, blood plasma of Prussian carp (Carassius gibelio, which was subjected to natural cold-acclimation, and purified protein antifreeze tmAFP isolated from larval mealworm (Tenebrio molitor, which also was subjected to natural cold-acclimation, were sued as modifiers. Findings. The results of the study demonstrated that qualitative and quantitative characteristics of the defrosted sperm, results of the incubation, rearing of embryos and larvae of Nyvky scaled carp (NLC, as well as fish culture parameters of produced young-of-the-year depended from on the composition of cryoprotective medium. The best results were demonstrated for the experimental groups, obtained with the use of the cryoprotective solution supplemented with purified antifreeze protein tmAFP. The larvae obtained from the sperm cryopreserved with the addition of cryoprotective medium TmAFP had better resistance to dehydration, surpassing the experimental groups obtained from the modifiers of Prussian carp plasma and cobamamide. There is a consolidation in the action of the related extracellular cryoprotectors, which were isolated from the cold-resistant organisms. And carp produced with their used demonstrated better performance during their rearing. The fact can be established that the manifestation of cryoselective effect depends on the integrity of sperm hereditary material and does not depend on the effect of extremely low temperatures on the cell membranes. Originality. There were the first experiments, where Prussian carp plasma and antifreeze protein tmAFP were used as extracellular cryoprotectors. Practical value. The modifications of composite

  1. Modeling of Rocket Fuel Heating and Cooling Processes in the Interior Receptacle Space of Ground-Based Systems

    Directory of Open Access Journals (Sweden)

    K. I. Denisova


    Full Text Available The propellant to fill the fuel tanks of the spacecraft, upper stages, and space rockets on technical and ground-based launch sites before fueling should be prepared to ensure many of its parameters, including temperature, in appropriate condition. Preparation of fuel temperature is arranged through heating and cooling the rocket propellants (RP in the tanks of fueling equipment. Processes of RP temperature preparation are the most energy-intensive and timeconsuming ones, which require that a choice of sustainable technologies and modes of cooling (heating RP provided by the ground-based equipment has been made through modeling of the RP [1] temperature preparation processes at the stage of design and operation of the groundbased fueling equipment.The RP temperature preparation in the tanks of the ground-based systems can be provided through the heat-exchangers built-in the internal space and being external with respect to the tank in which antifreeze, air or liquid nitrogen may be used as the heat transfer media. The papers [1-12], which note a promising use of the liquid nitrogen to cool PR, present schematic diagrams and modeling systems for the RP temperature preparation in the fueling equipment of the ground-based systems.We consider the RP temperature preparation using heat exchangers to be placed directly in RP tanks. Feeding the liquid nitrogen into heat exchanger with the antifreeze provides the cooling mode of PR while a heated air fed there does that of heating. The paper gives the systems of equations and results of modeling the processes of RP temperature preparation, and its estimated efficiency.The systems of equations of cooling and heating RP are derived on the assumption that the heat exchange between the fuel and the antifreeze, as well as between the storage tank and the environment is quasi-stationary.The paper presents calculation results of the fuel temperature in the tank, and coolant temperature in the heat exchanger, as

  2. Nano materials for efficiently lowering the freezing point of heat transfer nanofluids (United States)

    Hong, Haiping; Roy, Walter


    In this paper, we report, for the first time, the effect of the lowered freezing point in a 50% water / 50% antifreeze coolant (PAC) or 50% water / 50% ethylene glycol (EG) solution by the addition of carbon nanotubes and other particles. The experimental results indicated that the nano materials are much more efficient (hundreds fold) in lowering the freezing point than the regular ionic materials (e.g. NaCl). The possible explanation for this interesting phenomenon is the colligative property of fluid and relative small size of nano material. It is quite certain that the carbon nanotubes and metal oxide nano particles could be a wonderful candidate for the nano coolant application because they could not only increase the thermal conductivity, but also efficiently lower the freezing point of traditional coolants.

  3. Thermal hysteresis proteins. (United States)

    Barrett, J


    Extreme environments present a wealth of biochemical adaptations. Thermal hysteresis proteins (THPs) have been found in vertebrates, invertebrates, plants, bacteria and fungi and are able to depress the freezing point of water (in the presence of ice crystals) in a non-colligative manner by binding to the surface of nascent ice crystals. The THPs comprise a disparate group of proteins with a variety of tertiary structures and often no common sequence similarities or structural motifs. Different THPs bind to different faces of the ice crystal, and no single mechanism has been proposed to account for THP ice binding affinity and specificity. Experimentally THPs have been used in the cryopreservation of tissues and cells and to induce cold tolerance in freeze susceptible organisms. THPs represent a remarkable example of parallel and convergent evolution with different proteins being adapted for an anti-freeze role.

  4. Electro-suppression of water nano-droplets' solidification in no man's land: Electromagnetic fields' entropic trapping of supercooled water (United States)

    Nandi, Prithwish K.; Burnham, Christian J.; English, Niall J.


    Understanding water solidification, especially in "No Man's Land" (NML) (150 K < T < 235 K) is crucially important (e.g., upper-troposphere cloud processes) and challenging. A rather neglected aspect of tropospheric ice-crystallite formation is inevitably present electromagnetic fields' role. Here, we employ non-equilibrium molecular dynamics of aggressively quenched supercooled water nano-droplets in the gas phase under NML conditions, in externally applied electromagnetic (e/m) fields, elucidating significant differences between effects of static and oscillating fields: although static fields induce "electro-freezing," e/m fields exhibit the contrary - solidification inhibition. This anti-freeze action extends not only to crystal-ice formation but also restricts amorphisation, i.e., suppression of low-density amorphous ice which forms otherwise in zero-field NML environments. E/m-field applications maintain water in the deeply supercooled state in an "entropic trap," which is ripe for industrial impacts in cryo-freezing, etc.

  5. Recent Progress in Bionic Condensate Microdrop Self-Propelling Surfaces. (United States)

    Gong, Xiaojing; Gao, Xuefeng; Jiang, Lei


    Bionic condensate microdrop self-propelling (CMDSP) surfaces are attracting increased attention as novel, low-adhesivity superhydrophobic surfaces due to their value in fundamental research and technological innovation, e.g., for enhancing heat transfer, energy-effective antifreezing, and electrostatic energy harvesting. Here, the focus is on recent progress in bionic CMDSP surfaces. Metal-based CMDSP surfaces, which are the most promising in their respective fields, are highlighted for use in future applications. The selected topics are divided into four sections: biological prototypes, mechanism and construction rules, fabrication, and applications of metal-based CMDSP surfaces. Finally, the challenges and future development trends in bionic CMDSP surfaces are envisioned, especially the utilization of potential bionic inspiration in the design of more advanced CMDSP surfaces. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. The vasorelaxant mechanisms of methanol on isolated rat aortic rings: Involvement of ion channels and signal transduction pathways. (United States)

    Bai, Y; Zhang, Q; Yang, Z; Meng, Z; Zhao, Q


    It is reported that methanol is generally used as an industrial solvent, antifreeze, windshield washer fluid, cooking fuel and perfume. Methanol ingestion can lead to severe metabolic disturbances, blindness, or even death. So far, few studies about its negative effects on cardiovascular system have been reported. The purpose of this study was to determine the vasoactive effect of methanol and roles of ion channels and signal transduction pathways on isolated rat aorta. The results suggested that the mechanism of methanol-induced vasorelaxation at low concentrations (600 mM) was related to K ATP , voltage-dependent K + , big-conductance Ca 2+ -activated K + , L-type Ca 2+ channels as well as prostacyclin, protein kinase C, β-adrenoceptors pathways. In addition, methanol induced a dose-dependent inhibition of vasoconstrictions caused by calcium chloride, potassium chloride, or norepinephrine. Further work is needed to investigate the relative contribution of each channel and pathway in methanol-induced vasoactive effect.

  7. Temperatures and Heat Flows in a Soil Enclosing a Slinky Horizontal Heat Exchanger

    Directory of Open Access Journals (Sweden)

    Pavel Neuberger


    Full Text Available Temperature changes and heat flows in soils that host “slinky”-type horizontal heat exchangers are complex, but need to be understood if robust quantification of the thermal energy available to a ground-source heat pump is to be achieved. Of particular interest is the capacity of the thermal energy content of the soil to regenerate when the heat exchangers are not operating. Analysis of specific heat flows and the specific thermal energy regime within the soil, including that captured by the heat-exchangers, has been characterised by meticulous measurements. These reveal that high concentrations of antifreeze mix in the heat-transfer fluid of the heat exchanger have an adverse impact on heat flows discharged into the soil.

  8. Petroleum drilling and production operations in the Gulf of Mexico

    International Nuclear Information System (INIS)

    Fang, C.S.


    Decades of offshore and inland petroleum drilling and production in the Gulf of Mexico and on the Gulf Coast have provided the much needed energy and chemical feedstocks to the nation, and also have made an impact on the environment in the area. Our study showed deposits of contaminated sediment on the ocean floor around offshore platforms, old reserve pits, and dump sites next to many surface facilities and compressor stations. The substances found on the ocean floor and in dump sites are simple or emulsified mixtures of silt, hydrocarbons, and water. The cleaning of the ocean floor and pits is an economic and technical challenge. Hydrocarbons are from crude oil and chemical additions fro various operational necessities, including additions of biocides, corrosion inhibitors, antifreezes, and coagulants. When the new government regulations lower the allowable maximum total organic carbon level to the 50 ppm range, these hydrocarbons can no longer be ignored by drilling and production operators


    Directory of Open Access Journals (Sweden)

    VLASOV Vladimir Alexeevich


    Full Text Available The invention «Dispersion of Carbon Nanotubes (RU 2494961» can be used in production of modifying additives for construction materials. Dispersion of carbon nanotubes contains, mass %: carbon nanotubes 1–20; surface active agent – sodium chloride of sulfonated derived naphthalene 1–20; fumed silica 5–15; water – the rest. Dispersion can additionally contain ethylene glycol as antifreeze. Dispersion is steady in storage, it is soluble in water, provides increased strength of construction materials. Invention «Building Structures Reinforcement Composition (RU 2493337» can beused in construction to reinforce concrete, brick and masonry structures. Composition contains glass or basalt roving taken in quantity 90÷100 parts by weight, soaked in polymer binder based on epoxy taken in quantity 0,001÷1,5 parts by weight. This invention provides high resistance to operational load.

  10. The overwintering physiology of the emerald ash borer, Agrilus planipennis fairmaire (coleoptera: buprestidae). (United States)

    Crosthwaite, Jill C; Sobek, Stephanie; Lyons, D Barry; Bernards, Mark A; Sinclair, Brent J


    Ability to survive cold is an important factor in determining northern range limits of insects. The emerald ash borer (Agrilus planipennis) is an invasive beetle introduced from Asia that is causing extensive damage to ash trees in North America, but little is known about its cold tolerance. Herein, the cold tolerance strategy and mechanisms involved in the cold tolerance of the emerald ash borer were investigated, and seasonal changes in these mechanisms monitored. The majority of emerald ash borers survive winter as freeze-intolerant prepupae. In winter, A. planipennis prepupae have low supercooling points (approximately -30°C), which they achieve by accumulating high concentrations of glycerol (approximately 4M) in their body fluids and by the synthesis of antifreeze agents. Cuticular waxes reduce inoculation from external ice. This is the first comprehensive study of seasonal changes in cold tolerance in a buprestid beetle. 2010 Elsevier Ltd. All rights reserved.

  11. Synthesis of Degradable Poly(vinyl alcohol) by Radical Ring-Opening Copolymerization and Ice Recrystallization Inhibition Activity. (United States)

    Hedir, Guillaume; Stubbs, Christopher; Aston, Phillip; Dove, Andrew P; Gibson, Matthew I


    Poly(vinyl alcohol) (PVA) is the most active synthetic mimic of antifreeze proteins and has extremely high ice recrystallization inhibition (IRI) activity. Addition of PVA to cellular cryopreservation solutions increases the number of recovered viable cells due to its potent IRI, but it is intrinsically nondegradable in vivo . Here we report the synthesis, characterization, and IRI activity of PVA containing degradable ester linkages. Vinyl chloroacetate (VClAc) was copolymerized with 2-methylene-1,3-dioxepane (MDO) which undergoes radical ring-opening polymerization to install main-chain ester units. The use of the chloroacetate monomer enabled selective deacetylation with retention of esters within the polymer backbone. Quantitative IRI assays revealed that the MDO content had to be finely tuned to retain IRI activity, with higher loadings (24 mol %) resulting in complete loss of IRI activity. These degradable materials will help translate PVA, which is nontoxic and biocompatible, into a range of biomedical applications.

  12. Research on strength attenuation law of concrete in freezing - thawing environment (United States)

    Xiao, qianhui; Cao, zhiyuan; Li, qiang


    By rapid freezing and thawing method, the experiments of concrete have been 300 freeze-thaw cycles specimens in the water. The cubic compression strength value under different freeze-thaw cycles was measured. By analyzing the test results, the water-binder ratio of the concrete under freeze-thaw environments, fly ash and air entraining agent is selected dosage recommendations. The exponential attenuation prediction model and life prediction model of compression strength of concrete under freezing-thawing cycles considering the factors of water-binder ratio, fly ash content and air-entraining agent dosage were established. The model provides the basis for predicting the durability life of concrete under freezing-thawing environment. It also provides experimental basis and references for further research on concrete structures with antifreeze requirements.

  13. Thermal fluids in low temperature systems. Part 2

    Energy Technology Data Exchange (ETDEWEB)

    Lynde, P.G.; Yonkers, E.D. [Albert Kahn Associates, Inc., Detroit, MI (United States)


    This article focuses on the lifeblood of these systems, the thermal transfer fluid itself. Low-temperature heat-transfer fluids are used to condition engine fluids, test chambers, cooling fluids, or a combination of these in environmental test facilities. To meet the specific test criteria, these fluids may be required to maintain pumpability and function with thermal efficiency at temperatures as low as {minus}120 F. This article presents information related to heat-transfer fluids used in low-temperature cooling applications. Three general groups of fluids are discussed: water-based antifreezes (ethylene and propylene glycol solutions); chlorinated solvents (methylene chloride and trichloroethylene); organic and synthetic coolants (diethylbenzene, two forms of dimethylpolysiloxane, heavy naphtha hydrotreated, and citrus terpene).

  14. Solar heating and hot water system installed at Arlington Raquetball Club, Arlington, Virginia (United States)


    A solar space and water heating system is described. The solar energy system consists of 2,520 sq. ft. of flat plate solar collectors and a 4,000 gallon solar storage tank. The transfer medium in the forced closed loop is a nontoxic antifreeze solution (50 percent water, 50 percent propylene glycol). The service hot water system consists of a preheat coil (60 ft. of 1 1/4 in copper tubing) located in the upper third of the solar storage tank and a recirculation loop between the preheat coil and the existing electric water heaters. The space heating system consists of two separate water to air heat exchangers located in the ducts of the existing space heating/cooling systems. The heating water is supplied from the solar storage tank. Extracts from site files, specification references for solar modifications to existing building heating and hot water systems, and installation, operation and maintenance instructions are included.

  15. Eight fatalities due to drinking methanol-tainted alcohol in Pakistan: A case report

    Directory of Open Access Journals (Sweden)

    Humera Shafi


    Full Text Available Methanol has a widespread commercial use as a solvent in paints, varnishes, anti-freeze solutions and denaturant for ethanol. Exposure may occur due to accidental, suicidal ingestion or as a result of consuming adulterated liquor. Fatalities were reported in Pakistan in an incident after consuming methanol-tainted liquor. Postmortem specimens of eight deceased males, ages ranged from 16 to 40 yrs, were submitted for toxicological analysis. Presence of blurred vision, severe metabolic acidosis with decreased serum bicarbonate level, increased serum osmolality and mean anion gap before death strongly suggested methanol toxicity. Headspace gas chromatograph coupled to flame ionization detector was used to quantify volatiles in blood and stomach contents of victims. Lethal levels of methanol in addition to ethanol were detected. The most probable mechanism of methanol-related deaths was sudden cessation of respiration due to inhibition of cytochrome oxidase that led to histotoxic hypoxia.

  16. Ice cream structure modification by ice-binding proteins. (United States)

    Kaleda, Aleksei; Tsanev, Robert; Klesment, Tiina; Vilu, Raivo; Laos, Katrin


    Ice-binding proteins (IBPs), also known as antifreeze proteins, were added to ice cream to investigate their effect on structure and texture. Ice recrystallization inhibition was assessed in the ice cream mixes using a novel accelerated microscope assay and the ice cream microstructure was studied using an ice crystal dispersion method. It was found that adding recombinantly produced fish type III IBPs at a concentration 3 mg·L -1 made ice cream hard and crystalline with improved shape preservation during melting. Ice creams made with IBPs (both from winter rye, and type III IBP) had aggregates of ice crystals that entrapped pockets of the ice cream mixture in a rigid network. Larger individual ice crystals and no entrapment in control ice creams was observed. Based on these results a model of ice crystals aggregates formation in the presence of IBPs was proposed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Inhibition of nucleation and growth of ice by poly(vinyl alcohol) in vitrification solution. (United States)

    Wang, Hai-Yan; Inada, Takaaki; Funakoshi, Kunio; Lu, Shu-Shen


    Control of ice formation is crucial in cryopreservation of biological substances. Successful vitrification using several additives that inhibit ice nucleation in vitrification solutions has previously been reported. Among these additives, here we focused on a synthetic polymer, poly(vinyl alcohol) (PVA), and investigated the effects of PVA on nucleation and growth of ice in 35% (w/w) aqueous 1,2-propanediol solution by using a differential scanning calorimetry (DSC) system equipped with a cryomicroscope. First, the freezing temperature of the solution was measured using the DSC system, and then the change in ice fraction in the solution during cooling was evaluated based on images obtained using the cryomicroscope, at different concentrations of PVA between 0% and 3% (w/w). Based on the ice fraction, the change in residual solution concentration during cooling was also evaluated and then plotted on the state diagram of aqueous 1,2-propanediol solution. Results indicated that, when the partially glassy and partially frozen state was intentionally allowed, the addition of PVA effectively inhibited not only ice nucleation but also ice growth in the vitrification solution. The effect of PVA on ice growth in the vitrification solution was explained based on kinetic limitations mainly due to mass transport. The interfacial kinetics also might limit ice growth in the vitrification solution only when the ice growth rate decreased below a critical value. This coincides with the fact that PVA exhibits a unique antifreeze activity in the same manner as antifreeze proteins when ice growth rate is lower than a critical value.

  18. Molecular ecophysiology of Antarctic notothenioid fishes. (United States)

    Cheng, C-H Christina; Detrich, H William


    The notothenioid fishes of the Southern Ocean surrounding Antarctica are remarkable examples of organismal adaptation to extreme cold. Their evolution since the mid-Miocene in geographical isolation and a chronically cold marine environment has resulted in extreme stenothermality of the extant species. Given the unique thermal history of the notothenioids, one may ask what traits have been gained, and conversely, what characters have been lost through change in the information content of their genomes. Two dramatic changes that epitomize such evolutionary transformations are the gain of novel antifreeze proteins, which are obligatory for survival in icy seawater, by most notothenioids and the paradoxical loss of respiratory haemoproteins and red blood cells, normally deemed indispensable for vertebrate life, by the species of a highly derived notothenioid family, the icefishes. Here, we review recent advances in our understanding of these traits and their evolution and suggest future avenues of investigation. The formerly coherent paradigm of notothenioid freeze avoidance, developed from three decades of study of antifreeze glycoprotein (AFGP) based cold adaptation, now faces challenges stemming from the recent discovery of antifreeze-deficient, yet freeze-resistant, early notothenioid life stages and from definitive evidence that the liver is not the physiological source of AFGPs in notothenioid blood. The resolution of these intriguing observations is likely to reveal new physiological traits that are unique to the notothenioids. Similarly, the model of AFGP gene evolution from a notothenioid pancreatic trypsinogen-like gene precursor is being expanded and refined based on genome-level analyses of the linked AFGP loci and their ancestral precursors. Finally, the application of comparative genomics to study evolutionary change in the AFGP genotypes of cool-temperate notothenioids from sub-Antarctic habitats, where these genes are not necessary, will contribute to


    Directory of Open Access Journals (Sweden)

    L. P. Buchatsky


    Full Text Available The latest progress in biotechnology on fish aquaculture and different modern methods of investigations for increasing of fish productivity in aquaculture are analyzed. Except for the applied aspect, the use of modern biotechnological methods of investigations opens new possibilities for fundamental researches of sex-determining mechanisms, polyploidy, distant hybridization, and developmental biology of bony fishes. Review contains examples of utilizing modern biotechnology methods to obtain transgenic fishes with accelerated growth and for designing surrogate fishes. Methods for receiving unisexual shoals of salmon and sturgeon female fishes with the view of obtaining a large quantity of caviar, as well as receiving sterile (triploid fishes are analyzed. Great attention is given to androgenesis, particularly to disperm one, in connection with the problem of conserving rare and vanishing fish species using only sperm genetic material. Examples how distant hybrids may be obtained with the use of disperm androgenesis and alkylated DNA are given. Methods of obtaining fish primordium germ cells, recent developments in cultivation of fish stem cells and their use in biotechnology, as well as ones of transplantation of oogonium and spermatogonium to obtain surrogate fishes. The examples of successful experiments on spermatogonial xenotransplantation and characteristic of antifreezing fish proteins and also the prospect of their practical usage are given.

  20. Ice slurry accumulation

    Energy Technology Data Exchange (ETDEWEB)

    Christensen, K.G.; Kauffeld, M.


    More and more refrigeration systems are designed with secondary loops, thus reducing the refrigerant charge of the primary refrigeration plant. In order not to increase energy consumption by introducing a secondary refrigerant, alternatives to the well established single phase coolants (brines) and different concepts of the cooling plant have to be evaluated. Combining the use of ice-slurry - mixture of water, a freezing point depressing agent (antifreeze) and ice particles - as melting secondary refrigerant and the use of a cool storage makes it possible to build plants with secondary loops without increasing the energy consumption and investment. At the same time the operating costs can be kept at a lower level. The accumulation of ice-slurry is compared with other and more traditional storage systems. The method is evaluated and the potential in different applications is estimated. Aspects of practically use of ice-slurry has been examined in the laboratory at the Danish Technological Institute (DTI). This paper will include the final conclusions from this work concerning tank construction, agitator system, inlet, outlet and control. The work at DTI indicates that in some applications systems with ice-slurry and accumulation tanks have a great future. These applications are described by a varying load profile and a process temperature suiting the temperature of ice-slurry (-3 - -8/deg. C). (au)

  1. Neofunctionalization of zona pellucida proteins enhances freeze-prevention in the eggs of Antarctic notothenioids (United States)

    Cao, Lixue; Huang, Qiao; Wu, Zhichao; Cao, Dong-dong; Ma, Zhanling; Xu, Qianghua; Hu, Peng; Fu, Yanxia; Shen, Yu; Chan, Jiulin; Zhou, Cong-zhao; Zhai, Wanying; Chen, Liangbiao


    The mechanisms by which the eggs of the Antarctic notothenioid fishes avoid freezing are not fully understood. Zona pellucida proteins (ZPs) are constituents of the chorion which forms a protective matrix surrounding the egg. Here we report occurrence of freezing temperature-related gene expansion and acquisition of unusual ice melting-promoting (IMP) activity in a family of Antarctic notothenioid ZPs (AnnotoZPs). Members of AnnotoZPs are shown to bind with ice and non-colligatively depress the melting point of a solution in a range of 0.26 to 0.65 °C at a moderate concentration. Eggs of zebrafishes expressing an AnnotoZP transgene show improved melting point depression and enhanced survival in freezing conditions. Mutational analyses in a representative AnnotoZP indicate the ZP domain and patches of acidic residues are essential structures for the IMP activity. AnnotoZPs, therefore, represent a group of macromolecules that prevent freezing by a unique ZP–ice interaction mechanism distinct from the known antifreeze proteins. PMID:27698404

  2. Survival mechanisms of vertebrate ectotherms at subfreezing temperatures: applications in cryomedicine. (United States)

    Costanzo, J P; Lee, R E; DeVries, A L; Wang, T; Layne, J R


    Various marine fishes, amphibians, and reptiles survive at temperatures several degrees below the freezing point of their body fluids by virtue of adaptive mechanisms that promote freeze avoidance or freeze tolerance. Freezing is avoided by a colligative depression of the blood freezing point, supercooling of the body fluids, or the biosynthesis of unique antifreeze proteins that inhibit the propagation of ice within body fluids. Conversely, freeze tolerance is an adaptation for the survival of tissue freezing under ecologically relevant thermal and temporal conditions that is conferred by the biosynthesis of permeating carbohydrate cryoprotectants and an extensive dehydration of tissues and organs. Such cryoprotective responses, invoked by the onset of freezing, mitigate the osmotic stress associated with freeze-concentration of cytoplasm, attendant metabolic perturbations, and physical damage. Cryomedical research has historically relied on mammalian models for experimentation even though endotherms do not naturally experience subfreezing temperatures. Some vertebrate ectotherms have "solved" not only the problem of freezing individual tissues and organs, but also that of simultaneously freezing all organ systems. An emerging paradigm in cryomedicine is the application of principles governing natural cold hardiness to the development of protocols for the cryopreservation of mammalian tissues and organs.

  3. GEO-SOLE - A comparative hydrogeological and technical evaluation of heat transfer fluids for near-surface geothermal applications; GEO-SOLE. Vergleichende hydrogeologische und anlagentechnische Bewertung von Waermetraegerfluessigkeiten fuer oberflaechennahe geothermische Anwendungen

    Energy Technology Data Exchange (ETDEWEB)

    Koenigsdorff, Roland; Feuerstein, Philipp; Ryba, Meinhard [Hochschule Biberach (Germany). Inst. fuer Gebaeude- und Energiesysteme IGE


    Ground-coupled heat pump plants are commonly operated with mixtures of water and anti-freezing agents. Brines based on glycol are most frequently used, but there is also a variety of other organic and inorganic fluids available. Even the use of pure water is possible when frost-proof operation can be assured. The material properties of the fluids are well-known, but there exists no complete and consistent data basis to compare them, neither for the case of a leakage in the borehole heat exchanger's piping, nor with respect to the energy efficiency and operational characteristics of whole plants. The joint project GEO-SOLE of the Univerity of Applied Sciences Biberach (UASB) and the University Karlsruhe comprises experimental and theoretical investigations of various heat carrier fluids regarding their operational performance within heat pump plants and their characteristics when passing through the backfilling of boreholes and being dispersed within the ground. The results gained at UASB indicate that the thermal and energetic performance of the brines investigated show comparably small variations, i.e. not more then 5 % in the plant COP. The differences in the required borehole depth sum up to less than 10 % for turbulent flow. (orig.)

  4. NTP toxicology and carcinogensis studies of dipropylene glycol (CAS No. 25265-71-8) in F344/N rats and B6C3F1 mice (drinking water studies). (United States)


    Dipropylene glycol is found in antifreeze, air fresheners, cosmetic products, solvents, and plastics. We studied the effects of dipropylene glycol on male and female rats and mice to identify potential or cancer-related hazards to humans. We gave groups of 50 male and female mice drinking water containing dipropylene glycol at concentrations of 10,000, 20,000, or 40,000 parts per million (corresponding to 1%, 2%, or 4%) for two years. Male and female rats received concentrations of 2,500, 10,000, or 40,000 parts per million. Other groups received untreated water and were the control group. Tissues from more than 40 sites were examined for every animal. The groups of animals receiving 40,000 ppm dipropylene glycol weighed less than the control animals. All the make rats receiving 40,000 ppm dipropylene glycol died before the end of the study, mainly because of kidney disease. All the other animal group survived as well as the controls. No increase in tumor rates were seen in any of the groups of rats or mice. We conclude that dipropylene glycol did not cause cancer in male or female rats or mice. Exposure to dipropylene glycol did increase the rate and severity of kidney nephropathy and inflammation of the liver and salivary gland in male rats and some atrophy of the epithelial tissue of the nose in male and female rats.

  5. Professionalism: introduction

    Directory of Open Access Journals (Sweden)

    Raschke RA


    Full Text Available No abstract available. Article truncated at 150 words. An important event in my career occurred about 20 years ago, late on a Friday afternoon. I was scheduled on call in the ICU for the entire 72-hour weekend, and even though I was just getting started, I was already tired and in a lousy mood. At 5 PM, I got a consult to see a patient in the neuro ICU. He was a 34-year-old man who had attempted suicide by drinking ethylene glycol antifreeze after an argument with his girlfriend. He had initially stabilized from a medical standpoint, but then developed delayed-onset cerebral edema. The team that was taking care of him had unsuccessfully pursued all treatment options. After 8 days of effort, he remained in a deep coma, near brain death. Now, with nothing left to try, and no hope left for a good outcome, they were dumping responsibility onto me just in time for the weekend. I ...

  6. Transcriptome Analysis of Spartina pectinata in Response to Freezing Stress.

    Directory of Open Access Journals (Sweden)

    Gyoungju Nah

    Full Text Available Prairie cordgrass (Spartina pectinata, a perennial C4 grass native to the North American prairie, has several distinctive characteristics that potentially make it a model crop for production in stressful environments. However, little is known about the transcriptome dynamics of prairie cordgrass despite its unique freezing stress tolerance. Therefore, the purpose of this work was to explore the transcriptome dynamics of prairie cordgrass in response to freezing stress at -5°C for 5 min and 30 min. We used a RNA-sequencing method to assemble the S. pectinata leaf transcriptome and performed gene-expression profiling of the transcripts under freezing treatment. Six differentially expressed gene (DEG groups were categorized from the profiling. In addition, two major consecutive orders of gene expression were observed in response to freezing; the first being the acute up-regulation of genes involved in plasma membrane modification, calcium-mediated signaling, proteasome-related proteins, and transcription regulators (e.g., MYB and WRKY. The follow-up and second response was of genes involved in encoding the putative anti-freezing protein and the previously known DNA and cell-damage-repair proteins. Moreover, we identified the genes involved in epigenetic regulation and circadian-clock expression. Our results indicate that freezing response in S. pectinata reflects dynamic changes in rapid-time duration, as well as in metabolic, transcriptional, post-translational, and epigenetic regulation.

  7. Sustainable Carbon/Carbon Supercapacitors Operating Down to -40 °C in Aqueous Electrolyte Made with Cholinium Salt. (United States)

    Abbas, Qamar; Béguin, François


    Cholinium chloride at a concentration of 5 mol kg -1 in water is proposed as a low-cost and environmentally friendly aqueous electrolyte, enabling extension of the operating range of carbon/carbon supercapacitors (SCs) down to -40 °C. This solution has a pH close to neutrality (pH 6.1) and high conductivity of 88 mS cm -1 at 24 °C. The supercapacitors demonstrate a high capacitance of 126 F g -1 (per mass of one electrode) and long life span at voltages up to 1.5 V. At -40 °C, the carbon/carbon SCs display excellent electrochemical characteristics with only slightly reduced capacitance of 106 F g -1 and negligible ohmic losses. As compared to previous works, where antifreezing additives were introduced in traditional neutral electrolytes, the low solubility of the salt and related poor conductivity of the solution is no longer an issue, which makes cholinium salt aqueous solutions very promising for SCs operating at sub-ambient temperature conditions. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Dynamics of Physical and Physicochemical Properties of Urban Soils under the Effect of Ice-Melting Salts (United States)

    Azovtseva, N. A.; Smagin, A. V.


    Physical (water content, density, and air and water regimes) and physicochemical (electrical conductivity, pH, and SAR) properties of urban soils were investigated on test plots of Moscow to evaluate their dynamics under anthropogenic impact. The wilting point and the dependence of the capillary-sorption and total water potentials of the soil water content were determined in laboratory experiments with natural and artificially saline soil samples to evaluate the effect of salt antifreeze substances on water availability for plants under conditions of active application of deicing reagents. Seasonal dynamics of these parameters were investigated. It was found that electrolytes display a steady tendency for the accumulation and redistribution in the root zone rather than for their deep leaching despite humid climatic conditions in Moscow megalopolis. In summer, regular droughts result in drying of the root zone to critical values and to the concentration of electrolytes up to the values that make the total water potential of soil unsuitable for water uptake by roots. The key factor of soil degradation under the impact of electrolytes is the soil dispersity: the finer the texture, the higher the soil salinization and solonetzicity and the stronger irreversible changes in the soil water retention capacity and physical properties.

  9. Carbohydrate-Based Ice Recrystallization Inhibitors Increase Infectivity and Thermostability of Viral Vectors (United States)

    Ghobadloo, Shahrokh M.; Balcerzak, Anna K.; Gargaun, Ana; Muharemagic, Darija; Mironov, Gleb G.; Capicciotti, Chantelle J.; Briard, Jennie G.; Ben, Robert N.; Berezovski, Maxim V.


    The inability of vaccines to retain sufficient thermostability has been an obstacle to global vaccination programs. To address this major limitation, we utilized carbohydrate-based ice recrystallization inhibitors (IRIs) to eliminate the cold chain and stabilize the potency of Vaccinia virus (VV), Vesicular Stomatitis virus (VSV) and Herpes virus-1 (HSV-1). The impact of these IRIs was tested on the potency of the viral vectors using a plaque forming unit assay following room temperature storage, cryopreservation with successive freeze-thaw cycles and lyophilization. Viral potency after storage with all three conditions demonstrated that N-octyl-gluconamide (NOGlc) recovered the infectivity of shelf stored VV, 5.6 Log10 PFU mL-1 during 40 days, and HSV-1, 2.7 Log10 PFU mL-1 during 9 days. Carbon-linked antifreeze glycoprotein analogue ornithine-glycine-glycine-galactose (OGG-Gal) increases the recovery of VV and VSV more than 1 Log10 PFU mL-1 after 10 freeze-thaw cycles. In VSV, cryostorage with OGG-Gal maintains high infectivity and reduces temperature-induced aggregation of viral particles by 2 times that of the control. In total, OGG-Gal and NOGlc preserve virus potency during cryostorage. Remarkably, NOGlc has potential to eliminate the cold chain and permit room temperature storage of viral vectors.

  10. Effects of six substances on the growth and freeze-drying of Lactobacillus delbrueckii subsp. bulgaricus. (United States)

    Chen, He; Huang, Jie; Shi, Xiaoyu; Li, Yichao; Liu, Yu


    The efficacy of Lactobacillus delbrueckii subsp. bulgaricus as starter cultures for the dairy industry depends largely on the number of viable and active cells. Freeze-drying is the most convenient and successful method to preserve the bacterial cells. However, not all strains survived during freeze-drying. The effects of six substances including NaCl, sorbitol, mannitol, mannose, sodium glutamate, betaine added to the MRS medium on the growth and freeze-drying survival rate and viable counts of Lb. delbrueckii subsp. bulgaricus were studied through a single-factor test and Plackett-Burman design. Subsequently, the optimum freeze-drying conditions of Lb. delbrueckii subsp. bulgaricus were determined. Lb. delbrueckii subsp. bulgaricus survival rates were up to the maximum of 42.7%, 45.4%, 23.6%, while the concentrations of NaCl, sorbitol, sodium glutamate were 0.6%, 0.15%, 0.09%, respectively. In the optimum concentration, the viable counts in broth is 6.1, 6.9, 5.13 (×108 CFU/mL), respectively; the viable counts in freeze-drying power are 3.09, 5.2, 2.7 (×1010 CFU/g), respectively. Three antifreeze factors including NaCl, sorbitol, sodium glutamate have a positive effect on the growth and freeze-drying of Lb. delbrueckii subsp. bulgaricus. The results are beneficial for developing Lb. delbrueckii subsp. bulgaricus.

  11. Elevated CO{sub 2} and development of frost hardiness in Norway spruce (picea abies (L.) Karst.); Oekt CO{sub 2} og utvikling av frostherdighet i gran

    Energy Technology Data Exchange (ETDEWEB)

    Dalen, Lars Sandved


    This thesis discusses controlled laboratory experiments carried out to study the effects of CO{sub 2} pollution on Norwegian spruce. It was found that elevated CO{sub 2} increased height growth and biomass production. It slightly increased frost hardiness, but only at high nitrogen values. There was no evidence of adverse effects of elevated CO{sub 2} on the phenology of bud set and the development of frost hardiness. Although not statistically significant, there seemed to be a consistently higher concentration of soluble carbohydrates in one-season-old Norway spruce seedlings treated with elevated CO{sub 2}. This was not found in three-year-old seedlings grown in open top chambers, possibly indicating a down-regulation of photosynthesis or a transition from free to predetermined growth, and change in allocation of photosynthates with age. Treatment with high or low concentrations of CO{sub 2} and nitrogen fertilizer did not affect apoplastic chitinolytic activity during cold acclimation, nor were there any effects on antifreeze activity in these apoplastic extracts from cold acclimated needles. 149 refs., 21 figs., 8 tabs.

  12. Extremophiles and their application to veterinary medicine

    Directory of Open Access Journals (Sweden)

    Irwin Jane A


    Full Text Available Extremophiles are organisms that can grow and thrive in harsh conditions, e.g., extremes of temperature, pH, salinity, radiation, pressure and oxygen tension. Thermophilic, halophilic and radiation-resistant organisms are all microbes, some of which are able to withstand multiple extremes. Psychrophiles, or cold-loving organisms, include not only microbes, but fish that live in polar waters and animals that can withstand freezing. Extremophiles are structurally adapted at a molecular level to withstand these conditions. Thermophiles have particularly stable proteins and cell membranes, psychrophiles have flexible cellular proteins and membranes and/or antifreeze proteins, salt-resistant halophiles contain compatible solutes or high concentrations of inorganic ions, and acidophiles and alkaliphiles are able to pump ions to keep their internal pH close to neutrality. Their interest to veterinary medicine resides in their capacity to be pathogenic, and as sources of enzymes and other molecules for diagnostic and pharmaceutical purposes. In particular, thermostable DNA polymerases are a mainstay of PCR-based diagnostics.

  13. Hydrogeology along the southern boundary of the Hanford Site between the Yakima and Columbia Rivers, Washington

    International Nuclear Information System (INIS)

    Liikala, T.L.


    US Department of Energy (DOE) operations at the Hanford Site, located in southeastern Washington, have generated large volumes of hazardous and radioactive wastes since 1944. Some of the hazardous wastes were discharged to the ground in the 1100 and 3000 Areas, near the city of Richland. The specific waste types and quantities are unknown; however, they probably include battery acid, antifreeze, hydraulic fluids, waste oils, solvents, degreasers, paints, and paint thinners. Between the Yakima and Columbia rivers in support of future hazardous waste site investigations and ground-water and land-use management. The specific objectives were to collect and review existing hydrogeologic data for the study area and establish a water-level monitoring network; describe the regional and study area hydrogeology; develop a hydrogeologic conceptual model of the unconfined ground-water flow system beneath the study area, based on available data; describe the flow characteristics of the unconfined aquifer based on the spatial and temporal distribution of hydraulic head within the aquifer; use the results of this study to delineate additional data needs in support of future Remedial Investigation/Feasibility Studies (RI/FS), Fate and Transport modeling, Baseline Risk Assessments (BRA), and ground-water and land-use management

  14. Perplexing cooperative folding and stability of a low-sequence complexity, polyproline 2 protein lacking a hydrophobic core. (United States)

    Gates, Zachary P; Baxa, Michael C; Yu, Wookyung; Riback, Joshua A; Li, Hui; Roux, Benoît; Kent, Stephen B H; Sosnick, Tobin R


    The burial of hydrophobic side chains in a protein core generally is thought to be the major ingredient for stable, cooperative folding. Here, we show that, for the snow flea antifreeze protein (sfAFP), stability and cooperativity can occur without a hydrophobic core, and without α-helices or β-sheets. sfAFP has low sequence complexity with 46% glycine and an interior filled only with backbone H-bonds between six polyproline 2 (PP2) helices. However, the protein folds in a kinetically two-state manner and is moderately stable at room temperature. We believe that a major part of the stability arises from the unusual match between residue-level PP2 dihedral angle bias in the unfolded state and PP2 helical structure in the native state. Additional stabilizing factors that compensate for the dearth of hydrophobic burial include shorter and stronger H-bonds, and increased entropy in the folded state. These results extend our understanding of the origins of cooperativity and stability in protein folding, including the balance between solvent and polypeptide chain entropies.

  15. Inhibition of hydrate formation by kinetic inhibitors. Literature study; Inhibierung von Erdgashydraten durch kinetische Inhibitoren. Literaturstudie

    Energy Technology Data Exchange (ETDEWEB)

    Eberhardt, E.; Meyn, V.; Rahimian, I. [Institut fuer Erdoel- und Erdgasforschung, Clausthal-Zellerfeld (Germany)


    The aim of this study was to represent the state-of-the art of the inhibition of gas hydrates. Corresponding to recent publications the kinetic inhibition was considered in particular. Special inhibitors were validated using a set of criteria derived from different experimental test methods. Best results were obtained by the application of terpolymer VC-713 especially in relation to nucleation and crystal growth, followed by PVCap (polyvinylcaprolactame) and THI (threshold hydrate inhibitor), the chemical structure of which is derived from the antifreeze glycopeptids of antarcitc winter flounder. (orig.) [German] Die vorliegende Literaturstudie gibt den derzeitigen Stand der Kenntnis zur Inhibierung von Gashydraten wieder. Entsprechend der neueren Literatur wird insbesondere auf die kinetische Inhibierung eingegangen. Zur Beurteilung der verschiedenen Inhibitoren werden Bewertungskriterien zur Validierung der mit unterschiedlichen Untersuchungsmethoden erzielten experimentellen Ergebnisse angegeben. Anhand dieser Vorgehensweise zeigte sich, dass mit dem Terpolymer VC-713 die besten Ergebnisse, insbesondere im Hinblick auf Keimbildung und Wachstum, erzielt werden konnten. Sehr gute Ergebnisse wurden auch mit dem Polyvinylcaprolactam (PVCap) und den aus den Antigefrierpeptiden der antarktischen Winterflunder abgeleiteten Threshold Hydrate Inhibitoren (THI) erhalten. (orig.)

  16. Time-resolved laser-induced fluorescence system (United States)

    Bautista, F. J.; De la Rosa, J.; Gallegos, F. J.


    Fluorescence methods are being used increasingly in the measurement of species concentrations in gases, liquids and solids. Laser induced fluorescence is spontaneous emission from atoms or molecules that have been excited by laser radiation. Here we present a time resolved fluorescence instrument that consists of a 5 μJ Nitrogen laser (337.1 nm), a sample holder, a quartz optical fiber, a spectrometer, a PMT and a PC that allows the measurement of visible fluorescence spectra (350-750 nm). Time response of the system is approximately 5 ns. The instrument has been used in the measurement of colored bond paper, antifreeze, diesel, cochineal pigment and malignant tissues. The data acquisition was achieved through computer control of a digital oscilloscope (using General Purpose Interface Bus GPIB) and the spectrometer via serial (RS232). The instrument software provides a graphic interface that lets make some data acquisition tasks like finding fluorescence spectra, and fluorescence lifetimes. The software was developed using the Lab-View 6i graphic programming package and can be easily managed in order to add more functions to it.

  17. Inhibition of Ice Growth and Recrystallization by Zirconium Acetate and Zirconium Acetate Hydroxide (United States)

    Mizrahy, Ortal; Bar-Dolev, Maya; Guy, Shlomit; Braslavsky, Ido


    The control over ice crystal growth, melting, and shaping is important in a variety of fields, including cell and food preservation and ice templating for the production of composite materials. Control over ice growth remains a challenge in industry, and the demand for new cryoprotectants is high. Naturally occurring cryoprotectants, such as antifreeze proteins (AFPs), present one solution for modulating ice crystal growth; however, the production of AFPs is expensive and inefficient. These obstacles can be overcome by identifying synthetic substitutes with similar AFP properties. Zirconium acetate (ZRA) was recently found to induce the formation of hexagonal cavities in materials prepared by ice templating. Here, we continue this line of study and examine the effects of ZRA and a related compound, zirconium acetate hydroxide (ZRAH), on ice growth, shaping, and recrystallization. We found that the growth rate of ice crystals was significantly reduced in the presence of ZRA and ZRAH, and that solutions containing these compounds display a small degree of thermal hysteresis, depending on the solution pH. The compounds were found to inhibit recrystallization in a manner similar to that observed in the presence of AFPs. The favorable properties of ZRA and ZRAH suggest tremendous potential utility in industrial applications. PMID:23555701

  18. Ice Recrystallization in a Solution of a Cryoprotector and Its Inhibition by a Protein: Synchrotron X-Ray Diffraction Study. (United States)

    Zakharov, Boris; Fisyuk, Alexander; Fitch, Andy; Watier, Yves; Kostyuchenko, Anastasia; Varshney, Dushyant; Sztucki, Michael; Boldyreva, Elena; Shalaev, Evgenyi


    Ice formation and recrystallization is a key phenomenon in freezing and freeze-drying of pharmaceuticals and biopharmaceuticals. In this investigation, high-resolution synchrotron X-ray diffraction is used to quantify the extent of disorder of ice crystals in binary aqueous solutions of a cryoprotectant (sorbitol) and a protein, bovine serum albumin. Ice crystals in more dilute (10 wt%) solutions have lower level of microstrain and larger crystal domain size than these in more concentrated (40 wt%) solutions. Warming the sorbitol-water mixtures from 100 to 228 K resulted in partial ice melting, with simultaneous reduction in the microstrain and increase in crystallite size, that is, recrystallization. In contrast to sorbitol solutions, ice crystals in the BSA solutions preserved both the microstrain and smaller crystallite size on partial melting, demonstrating that BSA inhibits ice recrystallization. The results are consistent with BSA partitioning into quasi-liquid layer on ice crystals but not with a direct protein-ice interaction and protein sorption on ice surface. The study shows for the first time that a common (i.e., not-antifreeze) protein can have a major impact on ice recrystallization and also presents synchrotron X-ray diffraction as a unique tool for quantification of crystallinity and disorder in frozen aqueous systems. Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  19. Carbohydrate-Based Ice Recrystallization Inhibitors Increase Infectivity and Thermostability of Viral Vectors (United States)

    Ghobadloo, Shahrokh M.; Balcerzak, Anna K.; Gargaun, Ana; Muharemagic, Darija; Mironov, Gleb G.; Capicciotti, Chantelle J.; Briard, Jennie G.; Ben, Robert N.; Berezovski, Maxim V.


    The inability of vaccines to retain sufficient thermostability has been an obstacle to global vaccination programs. To address this major limitation, we utilized carbohydrate-based ice recrystallization inhibitors (IRIs) to eliminate the cold chain and stabilize the potency of Vaccinia virus (VV), Vesicular Stomatitis virus (VSV) and Herpes virus-1 (HSV-1). The impact of these IRIs was tested on the potency of the viral vectors using a plaque forming unit assay following room temperature storage, cryopreservation with successive freeze-thaw cycles and lyophilization. Viral potency after storage with all three conditions demonstrated that N-octyl-gluconamide (NOGlc) recovered the infectivity of shelf stored VV, 5.6 Log10 PFU mL−1 during 40 days, and HSV-1, 2.7 Log10 PFU mL−1 during 9 days. Carbon-linked antifreeze glycoprotein analogue ornithine-glycine-glycine-galactose (OGG-Gal) increases the recovery of VV and VSV more than 1 Log10 PFU mL−1 after 10 freeze-thaw cycles. In VSV, cryostorage with OGG-Gal maintains high infectivity and reduces temperature-induced aggregation of viral particles by 2 times that of the control. In total, OGG-Gal and NOGlc preserve virus potency during cryostorage. Remarkably, NOGlc has potential to eliminate the cold chain and permit room temperature storage of viral vectors. PMID:25078058

  20. Requirements and designs for Mars Rover RTGs (United States)

    Schock, A.; Sankarankandath, V.; Shirbacheh, M.


    A typical Rover mission, its requirements, the environment it imposes on the radionuclide thermoelectric generator (RTG), and a design approach for making the RTG operable in such an environment are described. Specific RTG designs for various thermoelectric element alternatives are presented. The results of studies show that current multifoil-insulated GPHS (general-purpose heat source) RTG and Mod-RTG designs can be modified to operate in an environment with an external atmosphere (e.g., Mars). This can be done while the helium generated by the fuel's alpha decay is vented to the external atmosphere. The use of novel selective vents and high-capacity getters is not required. The Rover RTGs can be built from standard, proven GPHS modules and standard SiGe unicouples or SiGe/GaP multicouples, using demonstrated thermoelectric material performance parameters. Rover's 500-W power requirement can be satisfied with two 250-W or four 125-W RTGs, whose sizes are compatible with currently envisaged Rover designs. An auxiliary cooling loop (e.g., water and antifreeze) will be required to cool the RTG while it is within the Rover's aeroshell during launch and transit to Mars.

  1. Extremophiles and their application to veterinary medicine (United States)


    Extremophiles are organisms that can grow and thrive in harsh conditions, e.g., extremes of temperature, pH, salinity, radiation, pressure and oxygen tension. Thermophilic, halophilic and radiation-resistant organisms are all microbes, some of which are able to withstand multiple extremes. Psychrophiles, or cold-loving organisms, include not only microbes, but fish that live in polar waters and animals that can withstand freezing. Extremophiles are structurally adapted at a molecular level to withstand these conditions. Thermophiles have particularly stable proteins and cell membranes, psychrophiles have flexible cellular proteins and membranes and/or antifreeze proteins, salt-resistant halophiles contain compatible solutes or high concentrations of inorganic ions, and acidophiles and alkaliphiles are able to pump ions to keep their internal pH close to neutrality. Their interest to veterinary medicine resides in their capacity to be pathogenic, and as sources of enzymes and other molecules for diagnostic and pharmaceutical purposes. In particular, thermostable DNA polymerases are a mainstay of PCR-based diagnostics. PMID:21851659

  2. Toxic optic neuropathy: An unusual cause

    Directory of Open Access Journals (Sweden)

    Hema L Ramkumar


    Full Text Available A 60-year-old woman with a history of chronic alcoholism and tobacco use presented with the complaint of a painless decrease in vision in both eyes. She lost vision first in the left eye then in the right eye. She admitted consuming at least one 16 ounce bottle of over the counter mouthwash daily and denied consumption of any other alcohols, methanol, or antifreeze. She stated that her vision had been continuing to deteriorate in both eyes. Her best-corrected visual acuity was 4/200 in each eye. Color vision was nil in each eye. Her pupils were sluggish bilaterally, and her optic discs were flat and hyperemic with peripapillary hemorrhages. Her visual fields revealed central scotomas bilaterally. The magnetic resonance imaging of the brain and lumbar puncture were within normal limits. Antinuclear antibody, human leukocyte antigen-B27 genotyping, and B12 were normal; serum thiamine was low. While continuing to ingest mouthwash, her vision decreased to count fingers at 2 feet, and maculopapillary bundle pallor developed. She was started on folate and thiamine supplementation. Once she discontinued mouthwash, her vision improved to 20/400 bilaterally, and her central scotomas improved. This case demonstrates an alcohol-induced toxic optic neuropathy from mouthwash ingestion with some visual recovery after discontinuation of the offending agent.

  3. Relevance of tidal heating on large TNOs (United States)

    Saxena, Prabal; Renaud, Joe P.; Henning, Wade G.; Jutzi, Martin; Hurford, Terry


    We examine the relevance of tidal heating for large Trans-Neptunian Objects, with a focus on its potential to melt and maintain layers of subsurface liquid water. Depending on their past orbital evolution, tidal heating may be an important part of the heat budget for a number of discovered and hypothetical TNO systems and may enable formation of, and increased access to, subsurface liquid water. Tidal heating induced by the process of despinning is found to be particularly able to compete with heating due to radionuclide decay in a number of different scenarios. In cases where radiogenic heating alone may establish subsurface conditions for liquid water, we focus on the extent by which tidal activity lifts the depth of such conditions closer to the surface. While it is common for strong tidal heating and long lived tides to be mutually exclusive, we find this is not always the case, and highlight when these two traits occur together. We find cases where TNO systems experience tidal heating that is a significant proportion of, or greater than radiogenic heating for periods ranging from100‧s of millions to a billion years. For subsurface oceans that contain a small antifreeze component, tidal heating due to very high initial spin states may enable liquid water to be preserved right up to the present day. Of particular interest is the Eris-Dysnomia system, which in those cases may exhibit extant cryovolcanism.

  4. Experimental study of the thermal characteristics of phase change slurries for active cooling

    International Nuclear Information System (INIS)

    Lu, W.; Tassou, S.A.


    Highlights: ► Tween 60 and hexadecanol can be employed to produce paraffin-in-water emulsions. ► Paraffin with longer carbon chain than the paraffin in the emulsion can act as nucleate agent to reduce supercooling. ► Increasing the quantity of paraffin increases the viscosity of the emulsion. ► Antifreeze and traces of thickener can cause a significant increase to the viscosity of the emulsion. ► Well prepared emulsions are stable with storage and thermal cycles. -- Abstract: Phase change materials (PCMs) are increasingly being used for thermal energy storage in buildings and industry to produce energy savings and reduce carbon dioxide emissions. PCM slurries are also being investigated for active thermal energy storage or as alternatives to conventional single phase fluids because they are pumpable and have advanced heat transport performance with phase change. The present study investigates several types of phase change materials for the preparation of PCM slurries which have potential for cooling applications. The thermophysical properties of paraffin in water emulsions, such as latent heat of fusion, melting and freezing temperature ranges, viscosity and the effect of surfactants, have been tested using appropriate experimental techniques. It has been identified that the use of small quantities of higher melting temperature paraffin and surfactants in the emulsion can reduce the effect of supercooling and increase the useful heat of fusion. However there are negative impacts on viscosity which should be considered in heat transport applications.

  5. The influence of calcium nitrate on setting and hardening rate of Portland cement concrete at different temperatures (United States)

    Kičaitė, A.; Pundienė, I.; Skripkiūnas, G.


    Calcium nitrate in mortars and concrete is used as a multifunctional additive: as set accelerator, plasticizer, long term strength enhancer and as antifreeze admixture. Used binding material and the amount of calcium nitrate, affect the characteristics of the concrete mixture and strength of hardened concrete. The setting time of the initial and the final binding at different temperatures of hardening (+ 20 °C and + 5 °C) of the pastes made of different cements (Portland cement CEM I 42.5 R and Portland limestone cement CEM II/A-LL 42.5 R) and various amounts of calcium nitrate from 1 % until 3 % were investigated. The effect of calcium nitrate on technological characteristics of concrete mixture (the consistency of the mixture, the density, and the amount of air in the mixture), on early concrete strength after 2 and 7 days, as well as on standard concrete strength after 28 days at different temperatures (at + 20 °C and + 5 °C) were analysed.

  6. Streamlined approach for environmental restoration work plan for Corrective Action Unit 126: Closure of aboveground storage tanks, Nevada Test Site, Nevada. Revision 1

    International Nuclear Information System (INIS)


    This plan addresses the closure of several aboveground storage tanks in Area 25 of the Nevada Test Site. The unit is currently identified as Corrective Action Unit 126 in the Federal Facility Agreement and Consent Order and is listed as having six Corrective Action Sites. This plan addresses the Streamlined Approach for Environmental Restoration closure for five of the six sites. Four of the CASs are located at the Engine Test Stand complex and one is located in the Central Support Area. The sites consist of aboveground tanks, two of which were used to store diesel fuel and one stored Nalcool (an antifreeze mixture). The remaining tanks were used as part of a water demineralization process and stored either sulfuric acid or sodium hydroxide, and one was used as a charcoal adsorption furnace. Closure will be completed by removal of the associated piping, tank supports and tanks using a front end loader, backhoe, and/or crane. When possible, the tanks will be salvaged as scrap metal. The piping that is not removed will be sealed using a cement grout

  7. Solar water heaters in China: A new day dawning

    International Nuclear Information System (INIS)

    Han Jingyi; Mol, Arthur P.J.; Lu, Yonglong


    Solar thermal utilization, especially the application of solar water heater technology, has developed rapidly in China in recent decades. Manufacturing and marketing developments have been especially strong in provinces such as Zhejiang, Shandong and Jiangsu. This paper takes Zhejiang, a relatively affluent province, as a case study area to assess the performance of solar water heater utilization in China. The study will focus on institutional setting, economic and technological performance, energy performance, and environmental and social impact. Results show that China has greatly increased solar water heater utilization, which has brought China great economic, environmental and social benefits. However, China is confronted with malfeasant market competition, technical flaws in solar water heater products and social conflict concerning solar water heater installation. For further development of the solar water heater, China should clarify the compulsory installation policy and include solar water heaters into the current 'Home Appliances Going to the Countryside' project; most of the widely used vacuum tube products should be replaced by flat plate products, and the technology improvement should focus on anti-freezing and water saving; the resources of solar water heater market should be consolidated and most of the OEM manufacturers should evolve to ODM and OBM enterprises.

  8. Ethylene Glycol and Metabolite Concentrations in Fatal Ethylene Glycol Poisonings. (United States)

    Viinamäki, Jenni; Sajantila, Antti; Ojanperä, Ilkka


    Ethylene glycol (EG) is used in antifreeze and other industrial products. It metabolizes to glycolic acid (GA) and oxalic acid (OX) that cause metabolic acidosis and are mainly responsible for the toxicity of EG. During 2010-2014, EG or GA was found in 25 postmortem cases in Finland. Of these cases, 21 were classified as fatal EG poisonings and 3 were classified as methanol (MeOH) poisonings. In this study, we report the concentrations of EG and GA in postmortem blood and urine samples of fatal EG or mixed MeOH/EG poisonings. In the fatal EG poisonings, the median EG and GA concentrations were 0.87 and 1.6 g/L in blood and 4.3 and 5.3 g/L in urine. The median urine-blood ratios were 3.8 and 3.1 for EG and GA. These results warrant the use of urine as a primary matrix for screening. In EG positive cases, the quantification of both EG and GA in blood is crucial as GA concentration appears to best indicate a fatal poisoning with an approximate threshold of 1.5 g/L. The measurement of urinary OX does not offer much additional value to toxic alcohol screening as it may originate from varying dietary conditions. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email:

  9. Delayed neurological sequelae from ethylene glycol, diethylene glycol and methanol poisonings. (United States)

    Reddy, Nandi J; Sudini, Madhuri; Lewis, Lionel D


    Ethylene glycol, diethylene glycol and methanol are widely available chemicals and are found in a variety of common household products including antifreeze, windshield washer fluid, brake fluid and lubricants. Following ingestion of these glycols and methanol, patients frequently develop an early neurological syndrome consisting of inebriation, ataxia, and if severe, seizures and coma. Though uncommon, a neurological syndrome may also develop as a delayed complication. Using Pub Med 438 references were identified of which 45 were relevant. Ethylene glycol poisoning has produced cranial nerve deficits (usually VII nerve dysfunction) after a delay of 5-20 days, Parkinsonism and cerebral edema. Diethylene glycol ingestion has been associated with the development of optic nerve injury, cranial nerve deficits, quadraparesis and peripheral neuropathy. Methanol poisoning has led to Parkinsonism and polyneuropathy. Oxalate crystal deposition likely causes the cranial neuropathies related to ethylene glycol and 2-hydroxyethoxyacetic acid is thought to be the causal moiety in cranial neuropathies resulting from diethylene glycol toxicity. Formic acid is implicated in the optic nerve damage associated with methanol. Uncommonly, delayed neurological syndromes may develop as complications of poisoning due to ethylene glycol, diethylene glycol and methanol; the onset of such neurological damage is often days or even weeks post-ingestion. Further research is required to explain why the facial nerve is the cranial nerve most commonly involved and why the basal ganglia are predisposed to injury.

  10. Detection of ethylene glycol - toward W51/e2 and G34.3+0.02 (United States)

    Lykke, Julie M.; Favre, Cécile


    Ethylene glycol (HOCH2CH2OH), also commenly known as antifreeze, is the reduced alcohol version of glycolaldehyde (CH2OHCHO). Glycoladehyde - the simplest possible aldehyde sugar (Marstokk and Møllendal 1973) - is the first intermediate step in the path toward forming more complex and biologically relevant molecules through the the formose reaction, which begins with formaldehyde (H2CO) and ends with the formation of sugars and ultimately ribose, the backbone of RNA (e.g., Larralde et al. 1995). The presence of glycolaldehyde is therefore an important indication that processes leading to biologically relevant molecules are taking place. It is however, still unclear as to how glycolaldehyde and ethylene glycol are formed in the ISM. It has been proposed that they share a common formation pathway through UV-irradiation of methanol (CH3OH) ices mixed with CO (Öberg et al. 2009). So far, ethylene glycol, in its lower energy con-former (g’Ga(CH2OH)2), has been detected toward SgrB2 (N) by Hollis et al. (2002), tentatively toward IRAS 16293-2422 (Jørgensen et al. 2012) and marginally by Kalenskii and Johansson (2010) toward W51 e1/e2. Here we present a firm detection of ethylene glycol toward W51/e2 as well as a first detection toward G34.3+0.02 at 1mm and 3mm using the IRAM 30m telescope.

  11. Reduction of energy essential to human life by 70%. Environment-friendly residence `ED-1` utilizing the sunlight effectively; Seikatsu ni hitsuyona energy no 70% sakugen. Taiyoko riyo no kankyo kyosei jutaku ED-1

    Energy Technology Data Exchange (ETDEWEB)



    This paper presents the environment-friendly residence `ED-1` utilizing the sunlight effectively. ED-1 of 97m{sup 2} in first floor area and 81m{sup 2} in second one has, on its roof, the photovoltaic power generation system, the solar hot water collector which supplies hot water by circulating heated antifreezing solution into a hot storage tank, and the solar hot wind collector which largely contributes to energy saving together with a hot room named an S unit on the first floor. The small S unit faced toward the south adjacent to a dining room is made of timber and thermal insulation glass. The S unit stores heat in a floor by taking the sunlight into a room in winter, while enhances a heating effect by discharging stored heat in nighttime. In some cases, duct circulation of heated wind is possible. Solar radiation of 90% can be removed by outside glass fiber screen of the S unit together with double glazing coated by special metal film. Roof material is superior in heat insulation, air tightness and energy saving, while floor material in sound insulation and heat storage. The target for reducing energy essential to human life by 70% was thus nearly achieved. 1 fig.

  12. Proteomics applied on plant abiotic stresses: role of heat shock proteins (HSP). (United States)

    Timperio, Anna Maria; Egidi, Maria Giulia; Zolla, Lello


    The most crucial function of plant cell is to respond against stress induced for self-defence. This defence is brought about by alteration in the pattern of gene expression: qualitative and quantitative changes in proteins are the result, leading to modulation of certain metabolic and defensive pathways. Abiotic stresses usually cause protein dysfunction. They have an ability to alter the levels of a number of proteins which may be soluble or structural in nature. Nowadays, in higher plants high-throughput protein identification has been made possible along with improved protein extraction, purification protocols and the development of genomic sequence databases for peptide mass matches. Thus, recent proteome analysis performed in the vegetal Kingdom has provided new dimensions to assess the changes in protein types and their expression levels under abiotic stress. As reported in this review, specific and novel proteins, protein-protein interactions and post-translational modifications have been identified, which play a role in signal transduction, anti-oxidative defence, anti-freezing, heat shock, metal binding etc. However, beside specific proteins production, plants respond to various stresses in a similar manner by producing heat shock proteins (HSPs), indicating a similarity in the plant's adaptive mechanisms; in plants, more than in animals, HSPs protect cells against many stresses. A relationship between ROS and HSP also seems to exist, corroborating the hypothesis that during the course of evolution, plants were able to achieve a high degree of control over ROS toxicity and are now using ROS as signalling molecules to induce HSPs.

  13. A novel GH secretagogue, A233, exhibits enhanced growth activity and innate immune system stimulation in teleosts fish. (United States)

    Martinez, Rebeca; Ubieta, Kenia; Herrera, Fidel; Forellat, Alina; Morales, Reynold; de la Nuez, Ania; Rodriguez, Rolando; Reyes, Osvaldo; Oliva, Ayme; Estrada, Mario P


    In teleosts fish, secretion of GH is regulated by several hypothalamic factors that are influenced by the physiological state of the animal. There is an interaction between immune and endocrine systems through hormones and cytokines. GH in fish is involved in many physiological processes that are not overtly growth related, such as saltwater osmoregulation, antifreeze synthesis, and the regulation of sexual maturation and immune functions. This study was conducted to characterize a decapeptide compound A233 (GKFDLSPEHQ) designed by molecular modeling to evaluate its function as a GH secretagogue (GHS). In pituitary cell culture, the peptide A233 induces GH secretion and it is also able to increase superoxide production in tilapia head-kidney leukocyte cultures. This effect is blocked by preincubation with the GHS receptor antagonist [d-Lys(3)]-GHRP6. Immunoneutralization of GH by addition of anti-tilapia GH monoclonal antibody blocked the stimulatory effect of A233 on superoxide production. These experiments propose a GH-mediated mechanism for the action of A233. The in vivo biological action of the decapeptide was also demonstrated for growth stimulation in goldfish and tilapia larvae (P<0.001). Superoxide dismutase levels, antiprotease activity, and lectin titer were enhanced in tilapia larvae treated with this novel molecule. The decapeptide A233 designed by molecular modeling is able to function as a GHS in teleosts and enhance parameters of the innate immune system in the fish larvae.

  14. Chemical diversity of organic volatiles among comets: An emerging taxonomy and implications for processes in the proto-planetary disk (United States)

    Mumma, Michael J.


    As messengers from the early Solar System, comets contain key information from the time of planet formation and even earlier some may contain material formed in our natal interstellar cloud. Along with water, the cometary nucleus contains ices of natural gases (CH4, C2H6), alcohols (CH3OH), acids (HCOOH), embalming fluid (H2CO), and even anti-freeze (ethylene glycol). Comets today contain some ices that vaporize at temperatures near absolute zero (CO, CH4), demonstrating that their compositions remain largely unchanged after 4.5 billion years. By comparing their chemical diversity, several distinct cometary classes have been identified but their specific relation to chemical gradients in the proto-planetary disk remains murky. How does the compositional diversity of comets relate to nebular processes such as chemical processing, radial migration, and dynamical scattering? No current reservoir holds a unique class, but their fractional abundance can test emerging dynamical models for origins of the scattered Kuiper disk, the Oort cloud, and the (proposed) main-belt comets. I will provide a simplified overview emphasizing what we are learning, current issues, and their relevance to the subject of this Symposium.

  15. Vertically Aligned Graphene Sheets Membrane for Highly Efficient Solar Thermal Generation of Clean Water. (United States)

    Zhang, Panpan; Li, Jing; Lv, Lingxiao; Zhao, Yang; Qu, Liangti


    Efficient utilization of solar energy for clean water is an attractive, renewable, and environment friendly way to solve the long-standing water crisis. For this task, we prepared the long-range vertically aligned graphene sheets membrane (VA-GSM) as the highly efficient solar thermal converter for generation of clean water. The VA-GSM was prepared by the antifreeze-assisted freezing technique we developed, which possessed the run-through channels facilitating the water transport, high light absorption capacity for excellent photothermal transduction, and the extraordinary stability in rigorous conditions. As a result, VA-GSM has achieved average water evaporation rates of 1.62 and 6.25 kg m -2 h -1 under 1 and 4 sun illumination with a superb solar thermal conversion efficiency of up to 86.5% and 94.2%, respectively, better than that of most carbon materials reported previously, which can efficiently produce the clean water from seawater, common wastewater, and even concentrated acid and/or alkali solutions.

  16. Corrosion problems with aqueous coolants, final report

    Energy Technology Data Exchange (ETDEWEB)

    Diegle, R B; Beavers, J A; Clifford, J E


    The results of a one year program to characterize corrosion of solar collector alloys in aqueous heat-transfer media are summarized. The program involved a literature review and a laboratory investigation of corrosion in uninhibited solutions. It consisted of three separate tasks, as follows: review of the state-of-the-art of solar collector corrosion processes; study of corrosion in multimetallic systems; and determination of interaction between different waters and chemical antifreeze additives. Task 1 involved a comprehensive review of published literature concerning corrosion under solar collector operating conditions. The reivew also incorporated data from related technologies, specifically, from research performed on automotive cooling systems, cooling towers, and heat exchangers. Task 2 consisted of determining the corrosion behavior of candidate alloys of construction for solar collectors in different types of aqueous coolants containing various concentrations of corrosive ionic species. Task 3 involved measuring the degradation rates of glycol-based heat-transfer media, and also evaluating the effects of degradation on the corrosion behavior of metallic collector materials.


    Directory of Open Access Journals (Sweden)

    Pedro Samuel Gomes Medeiros


    Full Text Available This paper makes a comparative analysis of the thermophysical properties of ice slurry with conventional single-phase secondary fluids used in thermal storage cooling systems. The ice slurry is a two-phase fluid consisting of water, antifreeze and ice crystals. It is a new technology that has shown great energy potential. In addition to transporting energy as a heat transfer fluid, it has thermal storage properties due to the presence of ice, storing coolness by latent heat of fusion. The single-phase fluids analyzed are water-NaCl and water-propylene glycol solutions, which also operate as carrier fluids in ice slurry. The presence of ice changes the thermophysical properties of aqueous solutions and a number of these properties were determined: density, thermal conductivity and dynamic viscosity. Data were obtained by software simulation. The results show that the presence of 10% by weight of ice provides a significant increase in thermal conductivity and dynamic viscosity, without causing changes in density. The rheological behavior of ice slurries, associated with its high viscosity, requires higher pumping power; however, this was not significant because higher thermal conductivity allows a lower mass flow rate without the use of larger pumps. Thus, the ice slurry ensures its high potential as a secondary fluid in thermal storage cooling systems, proving to be more efficient than single-phase secondary fluids.

  18. Solar water heating at Ikea's Montreal store; Chauffage solaire de l'eau courante au magasin Ikea de Montreal

    Energy Technology Data Exchange (ETDEWEB)

    Lagana, A.; Sonmor, K. [Ecovision experts-conseils inc., Montreal, PQ (Canada)


    The Ikea store in Montreal has integrated an energy efficient solar water heater into its building infrastructure. In June 2010, 30 fixed solar panels were installed on the roof to recuperate energy from the sun for hot water use in the restaurant and store facilities. Prior to this installation, the store was using natural gas for water heating purposes. During a sunny summer day, the solar water heater can provide 100 percent of the hot water needs of the store, even though it was designed to provide only one-third of the heating requirements. The panels are made of copper and painted with black titanium paint. A solution of water and antifreeze circulates in the panels. Digital controls are used at the pumps to control the temperature. The Ikea store uses approximately 3,100 gallons of water per day. The panels provide more than 60 MWh of energy, and represent a savings of 8440 cubic metres of natural gas, and a savings of 19.7 tons of carbon dioxide. The panels cost $1,300 and the return on investment is estimated to be 15.6 years. 3 figs.

  19. The Anion Gap is a Predictive Clinical Marker for Death in Patients with Acute Pesticide Intoxication. (United States)

    Lee, Sun-Hyo; Park, Samel; Lee, Jung-Won; Hwang, Il-Woong; Moon, Hyung-Jun; Kim, Ki-Hwan; Park, Su-Yeon; Gil, Hyo-Wook; Hong, Sae-Yong


    Pesticide formulation includes solvents (methanol and xylene) and antifreeze (ethylene glycol) whose metabolites are anions such as formic acid, hippuric acid, and oxalate. However, the effect of the anion gap on clinical outcome in acute pesticide intoxication requires clarification. In this prospective study, we compared the anion gap and other parameters between surviving versus deceased patients with acute pesticide intoxication. The following parameters were assessed in 1,058 patients with acute pesticide intoxication: blood chemistry (blood urea nitrogen, creatinine, glucose, lactic acid, liver enzymes, albumin, globulin, and urate), urinalysis (ketone bodies), arterial blood gas analysis, electrolytes (Na(+), K(+), Cl(-) HCO3 (-), Ca(++)), pesticide field of use, class, and ingestion amount, clinical outcome (death rate, length of hospital stay, length of intensive care unit stay, and seriousness of toxic symptoms), and the calculated anion gap. Among the 481 patients with a high anion gap, 52.2% had a blood pH in the physiologic range, 35.8% had metabolic acidosis, and 12.1% had acidemia. Age, anion gap, pesticide field of use, pesticide class, seriousness of symptoms (all P anion gap, and pesticide class were significant risk factors for death in a multiple logistic regression analysis (P anion gap is a significant risk factor for death, regardless of the accompanying acid-base balance status in patients with acute pesticide intoxication.

  20. Use and groundwater risk potential of additives in heat transfer fluids for borehole heat exchangers; Verwendung und Grundwassergefaehrdungspotenzial von Additiven in Waermetraegerfluessigkeiten fuer Erdwaermesonden

    Energy Technology Data Exchange (ETDEWEB)

    Ilieva, Dafina


    Ground based heat exchanger systems need to be evaluated in terms of potential effects on groundwater quality due to the risk of leakage of borehole heat exchanger fluids. The aim of this work was to identify the compounds which are present in additive mixtures and to investigate experimentally their biodegradability and effects on the biodegradation of the major organic component in borehole heat exchanger fluids. A data survey was carried out in cooperation with the State Ministry of the Environment Baden-Wuerttemberg, Germany to collect detailed information about the identity and application amounts of additives in borehole heat exchanger fluids. The survey revealed that numerous additives of various chemical classes and properties are used as corrosion inhibitors, alkalis, dyes, organic solvents, flavors, defoamers and surfactants. Furthermore, it was shown that glycols are among the most often applied antifreeze agents, the main component of the heat exchanger fluids. Based on the prioritization criteria (i) abundance in the borehole heat exchanger fluids, (ii) persistence, and (iii) mobility in the subsurface, the additives benzotriazole, tolyltriazole, 2-ethylhexanoate, benzoate and decane dicarboxylate were selected for further biodegradation experiments. The biodegradation experiments were carried out in batch systems with 60- or 70-m-deep sediments (sandstone or marl) as inoculum. The samples were taken during the installation of borehole heat exchanger systems at two different sites. The microcosms were conducted under oxic, denitrifying, iron- and sulfate-reducing as well as fermentative conditions at the presumed aquifer temperature of 12 C. The major component ethylene glycol was degraded under all conditions studied. The fastest biodegradation occurred under oxic and nitrate-reducing conditions (< 15 days). In all anoxic, nitrate free experiments with marl-sediment fermentation was the predominant process involved in the biodegradation of ethylene

  1. "Nanotechnology Enabled Advanced Industrial Heat Transfer Fluids"

    Energy Technology Data Exchange (ETDEWEB)

    Dr. Ganesh Skandan; Dr. Amit Singhal; Mr. Kenneth Eberts; Mr. Damian Sobrevilla; Prof. Jerry Shan; Stephen Tse; Toby Rossmann


    ABSTRACT Nanotechnology Enabled Advanced industrial Heat Transfer Fluids” Improving the efficiency of Industrial Heat Exchangers offers a great opportunity to improve overall process efficiencies in diverse industries such as pharmaceutical, materials manufacturing and food processing. The higher efficiencies can come in part from improved heat transfer during both cooling and heating of the material being processed. Additionally, there is great interest in enhancing the performance and reducing the weight of heat exchangers used in automotives in order to increase fuel efficiency. The goal of the Phase I program was to develop nanoparticle containing heat transfer fluids (e.g., antifreeze, water, silicone and hydrocarbon-based oils) that are used in transportation and in the chemical industry for heating, cooling and recovering waste heat. Much work has been done to date at investigating the potential use of nanoparticle-enhanced thermal fluids to improve heat transfer in heat exchangers. In most cases the effect in a commercial heat transfer fluid has been marginal at best. In the Phase I work, we demonstrated that the thermal conductivity, and hence heat transfer, of a fluid containing nanoparticles can be dramatically increased when subjected to an external influence. The increase in thermal conductivity was significantly larger than what is predicted by commonly used thermal models for two-phase materials. Additionally, the surface of the nanoparticles was engineered so as to have a minimal influence on the viscosity of the fluid. As a result, a nanoparticle-laden fluid was successfully developed that can lead to enhanced heat transfer in both industrial and automotive heat exchangers

  2. Control of glycerol production by rainbow smelt (Osmerus mordax) to provide freeze resistance and allow foraging at low winter temperatures. (United States)

    Driedzic, William R; Ewart, K Vanya


    The rainbow smelt (Osmerus mordax) is a small anadromous fish that actively feeds under the ice at temperatures as low as the freeze point of seawater. Freezing is avoided through the production of both non-colligative antifreeze protein (AFP) and glycerol that acts in a colligative manner. Glycerol is constantly lost across the gills and skin, thus glycerol production must continue on a sustained basis at low winter temperatures. AFP begins to accumulate in early fall while water temperatures are still high. Glycerol production is triggered when water temperatures decrease to about 5 degrees C. Glycerol levels rapidly increase with carbon flow from dihydroxyacetone phosphate (DHAP) to glycerol 3-phosphate (G3P) to glycerol. Glucose/glycogen serves as the initial carbon source for glycerol accumulation with amino acids contributing thereafter. The period of glycerol accumulation is associated with increases in GPDH mRNA and PEPCK mRNA followed by elevations in protein synthesis and enzyme activities. Plasma glycerol levels may reach in excess of 500 mM in winter. The high freeze resistance allows rainbow smelt to invade water of low temperature and forage for food. The lower the temperature, the higher the glycerol must be, and the higher the glycerol the greater the loss to the environment through diffusion. During the winter, rainbow smelt feed upon protein rich invertebrates with glycerol production being fueled in part by dietary amino acids via the gluconeogenic pathway. At winter temperatures, glycerol is quantitatively more important than AFP in providing freeze resistance of blood; however, the importance of AFPs to other tissues is yet to be assessed. Glycerol levels rapidly plummet in the spring when water temperature is still close to 0 degrees C. During this period, freeze resistance must be provided by AFP alone. Overall, the phenomenon of glycerol production by rainbow smelt reveals an elegant connection of biochemistry to ecology that allows this

  3. Reversible phosphorylation regulation of NADPH-linked polyol dehydrogenase in the freeze-avoiding gall moth, Epiblema scudderiana: role in glycerol metabolism. (United States)

    Holden, Helen A; Storey, Kenneth B


    Larvae of the goldenrod gall moth, Epiblema scudderiana, use a freeze avoidance strategy of cold hardiness to survive the winter. A key metabolic adaption that supports subzero survival is the accumulation of large amounts of glycerol as a colligative antifreeze. Production of glycerol relies on polyol dehydrogenase (PDH) which catalyzes the NADPH-dependent conversion of glyceraldehyde into glycerol. Kinetic analysis of PDH from E. scudderiana revealed significant changes in properties as a result of subzero temperature acclimation; the K(m) for glyceraldehyde in 5°C-acclimated larvae was 7.0 mM and doubled in - 15°C-exposed larvae. This change suggested that PDH is regulated by a state-dependent covalent modification. Indeed, high and low K(m) forms could be interconverted by incubating larval extracts in vitro under conditions that stimulated either endogenous protein kinases or protein phosphatases. Protein kinase incubations doubled the K(m) glyceraldehyde of the 5°C enzyme, whereas protein phosphatase incubations decreased the K(m) of the - 15°C enzyme by about 50%. PDH was purified by ion exchange and affinity chromatography steps and then subjected to electrophoresis. Staining with ProQ Diamond phosphoprotein stain showed a much higher phosphate content of PDH from - 15°C-acclimated larvae, a result that was further confirmed by immunoblotting that showed a much greater phosphoserine content on the - 15°C enzyme. These experiments established that PDH is regulated by state-dependent reversible phosphorylation in E. scudderiana and suggest that this regulatory mechanism makes a significant contribution to controlling the synthesis, maintenance, and degradation of glycerol pools over the winter months. © 2011 Wiley-Liss, Inc.

  4. Cold adaptation in marine organisms. (United States)

    Johnston, I A


    Animals from polar seas exhibit numerous so called resistance adaptations that serve to maintain homeostasis at low temperature and prevent lethal freezing injury. Specialization to temperatures at or below 0 degrees C is associated with an inability to survive at temperatures above 3-8 degrees C. Polar fish synthesize various types of glycoproteins or peptides to lower the freezing point of most extracellular fluid compartments in a non-colligative manner. Antifreeze production is seasonal in boreal species and is often initiated by environmental cues other than low temperature, particularly short day lengths. Most of the adaptations that enable intertidal invertebrates to survive freezing are associated with their ability to withstand ariel exposure. Unique adaptations for freezing avoidance include the synthesis of low molecular mass ice-nucleating proteins that control and induce extracellular ice-formation. Marine poikilotherms also exhibit a range of capacity adaptations that increase the rate of some physiological processes so as to partially compensate for the effects of low temperature. However, the rate of embryonic development in a diverse range of marine organisms shows no evidence of temperature compensation. This results in a significant lengthening of the time from fertilization to hatching in polar, relative to temperate, species. Some aspects of the physiology of polar marine species, such as low metabolic and slow growth rates, probably result from a combination of low temperature and other factors such as the highly seasonal nature of food supplies. Although neuromuscular function shows a partial capacity adaptation in Antarctic fish, maximum swimming speeds are lower than for temperate and tropical species, particularly for early stages in the life history.

  5. Contingent post-closure plan, hazardous waste management units at selected maintenance facilities, US Army National Training Center, Fort Irwin, California

    International Nuclear Information System (INIS)


    The National Training Center (NTC) at Fort Irwin, California, is a US Army training installation that provides tactical experience for battalion/task forces and squadrons in a mid- to high-intensity combat scenario. Through joint exercises with US Air Force and other services, the NTC also provides a data source for improvements of training doctrines, organization, and equipment. To meet the training and operational needs of the NTC, several maintenance facilities provide general and direct support for mechanical devices, equipment, and vehicles. Maintenance products used at these facilities include fuels, petroleum-based oils, lubricating grease, various degreasing solvents, antifreeze (ethylene glycol), transmission fluid, brake fluid, and hydraulic oil. Used or spent petroleum-based products generated at the maintenance facilities are temporarily accumulated in underground storage tanks (USTs), collected by the NTC hazardous waste management contractor (HAZCO), and stored at the Petroleum, Oil, and Lubricant (POL) Storage Facility, Building 630, until shipped off site to be recovered, reused, and/or reclaimed. Spent degreasing solvents and other hazardous wastes are containerized and stored on-base for up to 90 days at the NTC's Hazardous Waste Storage Facility, Building 703. The US Environmental Protection Agency (EPA) performed an inspection and reviewed the hazardous waste management operations of the NTC. Inspections indicated that the NTC had violated one or more requirements of Subtitle C of the Resource Conservation and Recovery Act (RCRA) and as a result of these violations was issued a Notice of Noncompliance, Notice of Necessity for Conference, and Proposed Compliance Schedule (NON) dated October 13, 1989. The following post-closure plan is the compliance-based approach for the NTC to respond to the regulatory violations cited in the NON

  6. The Effect of Early Frost Damage on the Penetration Resistance of Chloride Ion of NPP Concrete

    Energy Technology Data Exchange (ETDEWEB)

    Koh, Kyung Teak; Park, Chun Jin; Kim, Si Hwan; Ryu, Gum Sung [Korea Institute of Construction Technology, Goyang (Korea, Republic of)


    The specification for the nuclear power plant (NPP) structure construction specifies the conformity of the regulation ACI-306R in constructing the cold-weather concrete. According to the regulation with regard to the curing condition for cold weather concrete, the insulation curing of cold weather concrete should be appropriately performed under the environment of 5 .deg. C or more until the strength of 500 psi is developed. In addition, according to the regulations regarding the cold weather concrete in the domestic concrete specifications, the insulation curing should be performed until the strength development of 715 psi considering the safety factor indicated to the ACI regulation under the temperature of 5 .deg. C or more. According to the above-mentioned regulations, the NPP structure is required to develop the minimum strength of 715 psi or more and to maintain the important quality including strength development, early anti-freezing and duality under the cold weather condition. However, even though the early strength of 715 psi or more is secured under cold weather condition, if the structure is exposed to the continuous cold weather condition after the protection equipment including curing coat are removed, the structure's durability can go down compared to the concrete cured under the standard curing temperature condition in spring and fall, but the studies on this status still remain poor. Accordingly, this study tried to verify the adequacy of the insulation curing management standard, which is currently presented, in time of constructing the cold weather concrete, through reviewing the penetration resistance of chloride ion with considering the local characteristics of domestic NPP located at coastal areas after curing until the point of 715 psi, then exposing it to a certain cycle of freeze-thaw environment under the continuous cold weather condition

  7. Two cold-induced family 19 glycosyl hydrolases from cherimoya (Annona cherimola) fruit: an antifungal chitinase and a cold-adapted chitinase. (United States)

    Goñi, Oscar; Sanchez-Ballesta, María T; Merodio, Carmen; Escribano, María I


    Two cold-induced chitinases were isolated and purified from the mesocarp cherimoyas (Annona cherimola Mill.) and they were characterised as acidic endochitinases with a Mr of 24.79 and 47.77kDa (AChi24 and AChi48, respectively), both family 19 glycosyl hydrolases. These purified chitinases differed significantly in their biochemical and biophysical properties. While both enzymes had similar optimal acidic pH values, AChi24 was enzymatically active and stable at alkaline pH values, as well as displaying an optimal temperature of 45°C and moderate thermostability. Kinetic studies revealed a great catalytic efficiency of AChi24 for oligomeric and polymeric substrates. Conversely, AChi48 hydrolysis showed positive co-operativity that was associated to a mixture of different functional oligomeric states through weak transient protein interactions. The rise in the AChi48 kcat at increasing enzyme concentrations provided evidence of its oligomerisation. AChi48 chitinase was active and stable in a broad acidic pH range, and while it was relatively labile as temperatures increased, with an optimal temperature of 35°C, it retained about 50% of its maximal activity from 5 to 50°C. Thermodynamic characterisation reflected the high kcat of AChi48 and the remarkably lower ΔH(‡), ΔS(‡) and ΔG(‡) values at 5°C compared to AChi24, indicating that the hydrolytic activity of AChi48 was less thermodependent. In vitro functional studies revealed that AChi24 had a strong antifungal defence potential against Botrytis cinerea, whereas they displayed no cryoprotective or antifreeze activity. Hence, based on biochemical, thermodynamic and functional data, this study demonstrates that two acidic endochitinases are induced at low temperatures in a subtropical fruit, and that one of them acts in an oligomeric cold-adapted manner. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Immunochemical studies on the N-acetyllactosamine beta-(1----6)-linked trisaccharide specificity of Ricinus communis agglutinin. (United States)

    Wu, A M; Sugii, S; Gruezo, F G; Kabat, E A


    The combining site of Ricinus communis agglutinin (RCA1) was studied by quantitative precipitin and precipitin inhibition assays. Of 31 complex carbohydrates tested, all except active and inactive antifreeze glycoproteins, Streptococcus group C polysaccharide, and native rat salivary glycoprotein, reacted strongly, and 22 completely precipitated the lectin, indicating that RCA1 has both a broad range of affinity and a low solubility of its carbohydrate-bound complex. Of the monosaccharides and glycosides tested for inhibition of precipitation, p-nitrophenyl beta-D-galactopyranoside was the best. It was about 6.4 times better than methyl beta-D-galactopyranoside. The beta anomer of glycosides of D-galactose was much more potent than the corresponding alpha anomer. Among the oligosaccharides tested, beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----6)-D-Gal was the best inhibitor, which was approximately 2/3 as active as p-nitrophenyl beta-D-galactopyranoside. It was approximately 1.4 times as active as beta-D-Gal-(1----4)-D-GlcNAc (N-acetyllactosamine), twice as active as beta-D-Gal-(1----3)-D-GlcNAc, and 4.5 times more active than lacto-N-tetraose. From the results, it can be concluded that; (a) hydrophobic interaction is important for binding; (b) the combining site of this lectin is at least as large as a trisaccharide; and (c) of the compounds studied, the trisaccharide beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----6)-D-Gal was the most complementary to the human blood group I Ma determinant beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----6)-D-Gal.

  9. Thermal acclimation in rainbow smelt, Osmerus mordax, leads to faster myotomal muscle contractile properties and improved swimming performance

    Directory of Open Access Journals (Sweden)

    John R. Woytanowski


    Rainbow smelt (Osmerus mordax display an impressive ability to acclimate to very cold water temperatures. These fish express both anti-freeze proteins and glycerol in their plasma, liver, muscle and other tissues to avoid freezing at sub-zero temperatures. Maintenance of glycerol levels requires active feeding in very cold water. To understand how these fish can maintain activity at cold temperatures, we explored thermal acclimation by the myotomal muscle of smelt exposed to cold water. We hypothesized that cold-acclimated fish would show enhanced swimming ability due to shifts in muscle contractile properties. We also predicted that shifts in swimming performance would be associated with changes in the expression patterns of muscle proteins such as parvalbumin (PV and myosin heavy chain (MyHC. Swimming studies show significantly faster swimming by smelt acclimated to 5°C compared to fish acclimated to 20°C when tested at a common test temperature of 10°C. The cold-acclimated fish also had faster muscle contractile properties, such as a maximum shortening velocity (Vmax almost double that of warm-acclimated fish at the same test temperature. Cold-acclimation is associated with a modest increase in PV levels in the swimming muscle. Fluorescence microscopy using anti-MyHC antibodies suggests that MyHC expression in the myotomal muscle may shift in response to exposure to cold water. The complex set of physiological responses that comprise cold-acclimation in smelt includes modifications in muscle function to permit active locomotion in cold water.

  10. Simple and Efficient Trap for Bark and Ambrosia Beetles (Coleoptera: Curculionidae) to Facilitate Invasive Species Monitoring and Citizen Involvement. (United States)

    Steininger, M S; Hulcr, J; Šigut, M; Lucky, A


    Bark and ambrosia beetles (Coleoptera: Curculionidae: Scolytinae & Platypodinae) are among the most damaging forest pests worldwide, and monitoring is essential to damage prevention. Unfortunately, traps and attractants that are currently used are costly, and agencies rely on limited field personnel for deployment. The situation can be greatly aided by 1) the development of cost-effective trapping techniques, and 2) distribution of the effort through the Citizen Science approach. The goal of this study was to test a simple, effective trap that can be made and deployed by anyone interested in collecting bark and ambrosia beetles. Three trap types made from 2-liter soda bottles and, separately, four attractants were compared. Simple, one-window traps performed comparably at capturing species in traps painted or with multiple windows. A comparison of attractants in two-window traps found that 95% ethanol attracted the highest number of species but that Purell hand sanitizer (70% ethanol) and then Germ-X hand sanitizer (63% ethanol) were also effective. A perforated zip-top plastic bag containing Purell hanging over a trap filled with automobile antifreeze attracted the fewest species and individual specimens. Overall, >4,500 bark and ambrosia beetles, including 30 species were captured, representing a third of the regional species diversity. More than three quarters of the specimens were nonnative, representing nearly half of the known regional exotic species. These results suggest that simple one-window soda bottle traps baited with ethanol-based hand sanitizer will be effective and inexpensive tools for large-scale monitoring of bark and ambrosia beetles. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email:

  11. The Pharmacological Properties and Therapeutic Use of Apomorphine

    Directory of Open Access Journals (Sweden)

    Samo Ribarič


    Full Text Available Apomorphine (APO is an aporphine derivative used in human and veterinary medicine. APO activates D1, D2S, D2L, D3, D4, and D5 receptors (and is thus classified as a non-selective dopamine agonist, serotonin receptors (5HT1A, 5HT2A, 5HT2B, and 5HT2C, and α-adrenergic receptors (α1B, α1D, α2A, α2B, and α2C. In veterinary medicine, APO is used to induce vomiting in dogs, an important early treatment for some common orally ingested poisons (e.g., anti-freeze or insecticides. In human medicine, it has been used in a variety of treatments ranging from the treatment of addiction (i.e., to heroin, alcohol or cigarettes, for treatment of erectile dysfunction in males and hypoactive sexual desire disorder in females to the treatment of patients with Parkinson's disease (PD. Currently, APO is used in patients with advanced PD, for the treatment of persistent and disabling motor fluctuations which do not respond to levodopa or other dopamine agonists, either on its own or in combination with deep brain stimulation. Recently, a new and potentially important therapeutic role for APO in the treatment of Alzheimer’s disease has been suggested; APO seems to stimulate Ab catabolism in an animal model and cell culture, thus reducing the rate of Ab oligomerisation and consequent neural cell death.

  12. Information draft on the development of air standards for toluene

    Energy Technology Data Exchange (ETDEWEB)



    Toluene is a colorless, volatile liquid with a benzene-like odour. Its predominant use is in the production of benzene, as an octane enhancer in gasoline, as a solvent in aerosol spray paints, wall paints, lacquers, inks, adhesives, resins, and in such consumer products as spot removers, paint strippers, cosmetics, perfumes and antifreezes. Approximately 150 Ontario industrial sources reported toluene releases to the air totaling 4,245 to 5,300 tonnes during the reporting years from 1993 to 1996, making toluene one of the top pollutants by release quantities in Ontario and Canada for all these years. It is absorbed via the lungs and the gastrointestinal tract, both in humans and animals. Once absorbed, it tends to accumulate in the fatty tissues, and in vascularized tissues such as nerve cells and brain tissue. Toluene adversely affects the central nervous system (CNS) of humans and experimental animals. Observed symptoms in exposed humans range from decrease in psychometric performance, to headache, intoxication, convulsions, narcosis and death. Health Canada concluded that toluene is unlikely to be carcinogenic, although the available data is insufficient for definite classification. Ontario has 24-hour ambient air quality criterion and a half-hour Point of Impingement standard for toluene of 2,000 microgram/cubic meter, based on odour effects. The US Environmental Protection Agency inhalation reference concentration (also adopted by most of the American states) is 400 microgram/cubic meter. The WHO recommended a guideline value of 7500 microgram/cubic meter. Health Canada And Environment Canada established a tolerable concentration of 3750 microgram/cubic meter. 69 refs., 2 tabs., appendix

  13. Study of the mechanism of a kinetic inhibitor on the crystallization of methane hydrate; Etude du mecanisme d'action d'un inhibiteur cinetique sur la cristallisation de l'hydrate de methane

    Energy Technology Data Exchange (ETDEWEB)

    Pic, J.St.


    In the offshore exploitation of liquid fuels, problems of line plugging often occur, especially due to gas hydrates crystallization. At the present time, operators resort to antifreeze additives, which efficiency is defeated either by harder operating conditions or by a more severe environmental legislation. So research recently shifted towards a new class of 'low dosage inhibitors'. In order to understand the influence of such additives, we designed a high pressure reactor, fitted with a liquid injection device and an in situ turbidimetric sensor. Access to both the particle size distribution of the suspension during the first stages of crystallization, and the total gas consumption, allows us to characterize the kinetics of methane hydration formation. First, we developed an original experimental procedure, which generates an initial 'breeding' of the solution, and thus improves the mastering of nucleation. The induction time then becomes one of the relevant parameters to investigate the performance of inhibitors. Afterwards, we performed a first series of experiments which allowed us to determine the influence of the operating conditions (pressure and stirring) on the evolution of the particle size distribution, in the absence of additives. Then, we pointed out the inhibiting effect of a model kinetic inhibitor, polyvinylpyrrolidone. When dissolved in the solution before crystallization occurs, it increases the induction delay, decreases the gas consumption rate and also slows down the birth of new particles for several hours. On the contrary, when injected in the medium during crystallization, this polymer no more affects the reaction kinetics. At last, we raise the bases for a modelling, taking into account the elementary crystallization processes of nucleation, growth and particles agglomeration. A parametric study has been confronted to the experimental data. It enables us to suggest hypotheses regarding the effect of gas hydrates kinetic

  14. Rainbow smelt: the unusual case of cryoprotection by sustained glycerol production in an aquatic animal. (United States)

    Driedzic, William R


    Rainbow smelt flourish at -1.8 °C, the freezing point of sea water. An antifreeze protein contributes to freeze point depression but, more importantly, cryoprotection is due to an elevation in osmotic pressure, by the accumulation of glycerol. The lower the water temperature, the higher the plasma glycerol with levels recorded as high as 400 mmol l(-1). Glycerol freely diffuses out in direct relation to the glycerol concentration and fish may lose as much as 15% of their glycerol reserve per day. Glycerol levels decrease from a maximum in February/March while water temperature is still sub-zero. The decrease in glycerol may respond to a photoperiod signal as opposed to initiation which is triggered by low temperature. The initial increase in glycerol level is supported by liver glycogen but high sustained glycerol level is dependent upon dietary carbohydrate and protein. The metabolic pathways leading to glycerol involve flux from glycogen/glucose to the level of dihydroxyacetone phosphate (DHAP) via the initial part of glycolysis and from amino acids via a truncated gluconeogenesis again to the level of DHAP. DHAP in turn is converted to glycerol 3-phosphate (G3P) and then directly to glycerol. The key to directing DHAP to G3P is a highly active glycerol 3-P dehydrogenase. G3P is converted directly to glycerol via G3P phosphatase, the rate-limiting step in the process. The transition to glycerol production is associated with increased activities of enzymes at key loci in the top part of glycogenolysis/glycolysis. Curtailment of the final section of glycolysis may reside at the level of pyruvate oxidation with an inactivation of pyruvate dehydrogenase (PDH) driven by increased levels of PDH kinase. Enzymes associated with amino acid trafficking are elevated as is the pivotal enzyme phosphoenolpyruvate carboxykinase.

  15. Anti-ice nucleating activity of polyphenol compounds against silver iodide. (United States)

    Koyama, Toshie; Inada, Takaaki; Kuwabara, Chikako; Arakawa, Keita; Fujikawa, Seizo


    Freeze-avoiding organisms survive sub-zero temperatures without freezing in several ways, such as removal of ice nucleating agents (INAs), production of polyols, and dehydration. Another way is production of anti-ice nucleating agents (anti-INAs), such as has been reported for several antifreeze proteins (AFPs) and polyphenols, that inhibit ice nucleation by inactivating INAs. In this study, the anti-ice nucleating activity of five polyphenol compounds, including flavonoid and tannin compounds of both biological and synthetic origin, against silver iodide (AgI) was examined by measuring the ice nucleation temperature in emulsified polyphenol solutions containing AgI particles. The emulsified solutions eliminated the influence of contamination by unidentified INAs, thus enabling examination of the anti-ice nucleating activity of the polyphenols against AgI alone. Results showed that all five polyphenol compounds used here have anti-ice nucleating activities that are unique compared with other known anti-INAs, such as fish AFPs (type I and III) and synthetic polymers (poly(vinyl alcohol), poly(vinylpyrrolidone) and poly(ethylene glycol)). All five polyphenols completely inactivated the ice nucleating activity of AgI even at relatively low temperatures, and the first ice nucleation event was observed at temperatures between -14.1 and -19.4°C, compared with between -8.6 and -11.8°C for the fish AFPs and three synthetic polymers. These anti-ice nucleating activities of the polyphenols at such low temperatures are promising properties for practical applications where freezing should be prevented. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Physical Conditions and Exobiology Potential of Icy Satellites of the Giant Planets (United States)

    Simakov, M. B.


    All giant planets of the Solar system have a big number of satellites. A small part of them consist very large bodies, quite comparable to planets of terrestrial type, but including very significant share of water ice. Galileo spacecraft has given indications, primarily from magnetometer and gravity data, of the possibility that three of Jupiter's four large moons, Europa, Ganymede and Callisto have internal oceans. Formation of such satellites is a natural phenomenon, and satellite systems definitely should exist at extrasolar planets. The most recent models of the icy satellites interior lead to the conclusion that a substantial liquid layer exists today under relatively thin ice cover inside. The putative internal water ocean provide some exobiological niches on these bodies. We can see all conditions needed for origin and evolution of biosphere - liquid water, complex organic chemistry and energy sources for support of biological processes - are on the moons. The existing of liquid water ocean within icy world can be consequences of the physical properties of water ice, and they neither require the addition of antifreeze substances nor any other special conditions. On Earth life exists in all niches where water exists in liquid form for at least a portion of the year. Possible metabolic processes, such as nitrate/nitrite reduction, sulfate reduction and methanogenesis could be suggested for internal oceans of Titan and Jovanian satellites. Excreted products of the primary chemoautotrophic organisms could serve as a source for other types of microorganisms (heterotrophes). Subglacial life may be widespread among such planetary bodies as satellites of extrasolar giant planets, detected in our Galaxy.

  17. Transient freezing behavior in photophobic responses of Euglena gracilis investigated in a microfluidic device. (United States)

    Ozasa, Kazunari; Lee, Jeesoo; Song, Simon; Maeda, Mizuo


    We found that the transient freezing behavior in photophobic responses of Euglena gracilis is a good indicator of the metabolic status of the cells. The transient blue light photophobic responses of E. gracilis cells were investigated on-chip using a new measurement, 'trace momentum' (TM), to evaluate their swimming activity quantitatively in real time. When blue light of intensity >30 mW cm(-2) was repeatedly switched on and off, a large negative spike in the TM was observed at the onset of the 'blue-light-off' phase. Single-cell trace analysis at a blue light intensity of 40 mW cm(-2) showed that 48% (on average, n = 15) of tumbling Euglena cells ceased activity ('freezing') for 2-30 s at the onset of blue-light-off before commencing forward motion in a straight line (termed 'straightforward swimming'), while 45% smoothly commenced straightforward swimming without delay. The proportion of freezing Euglena cells depended on the blue light intensity (only 20% at 20 mW cm(-2)). When the cells were stimulated by four blue light pulses at the higher intensity, without pre-exposure, the transient freezing behavior was more prominent but, on repeating the stimuli after an 80 min interval in red light, the same cells did not freeze. This shows that the metabolism of the cells had changed to anti-freezing during the interval. The relationship between the interval time with/without light irradiation and the blue light adaptation was elucidated experimentally. The origin of the freezing behavior is considered to be a shortage of a metabolic substance that promotes smooth switching of flagellum movement from in situ rotation mode to a straightforward swimming mode. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email:

  18. Common protein sequence signatures associate with Sclerotinia borealis lifestyle and secretion in fungal pathogens of the Sclerotiniaceae. (United States)

    Badet, Thomas; Peyraud, Rémi; Raffaele, Sylvain


    Fungal plant pathogens produce secreted proteins adapted to function outside fungal cells to facilitate colonization of their hosts. In many cases such as for fungi from the Sclerotiniaceae family the repertoire and function of secreted proteins remains elusive. In the Sclerotiniaceae, whereas Sclerotinia sclerotiorum and Botrytis cinerea are cosmopolitan broad host-range plant pathogens, Sclerotinia borealis has a psychrophilic lifestyle with a low optimal growth temperature, a narrow host range and geographic distribution. To spread successfully, S. borealis must synthesize proteins adapted to function in its specific environment. The search for signatures of adaptation to S. borealis lifestyle may therefore help revealing proteins critical for colonization of the environment by Sclerotiniaceae fungi. Here, we analyzed amino acids usage and intrinsic protein disorder in alignments of groups of orthologous proteins from the three Sclerotiniaceae species. We found that enrichment in Thr, depletion in Glu and Lys, and low disorder frequency in hot loops are significantly associated with S. borealis proteins. We designed an index to report bias in these properties and found that high index proteins were enriched among secreted proteins in the three Sclerotiniaceae fungi. High index proteins were also enriched in function associated with plant colonization in S. borealis, and in in planta-induced genes in S. sclerotiorum. We highlight a novel putative antifreeze protein and a novel putative lytic polysaccharide monooxygenase identified through our pipeline as candidate proteins involved in colonization of the environment. Our findings suggest that similar protein signatures associate with S. borealis lifestyle and with secretion in the Sclerotiniaceae. These signatures may be useful for identifying proteins of interest as targets for the management of plant diseases.

  19. Common protein sequence signatures associate with Sclerotinia borealis lifestyle and secretion in fungal pathogens of the Sclerotiniaceae

    Directory of Open Access Journals (Sweden)

    Thomas eBadet


    Full Text Available Fungal plant pathogens produce secreted proteins adapted to function outside fungal cells to facilitate colonization of their hosts. In many cases such as for fungi from the Sclerotiniaceae family the repertoire and function of secreted proteins remains elusive. In the Sclerotiniaceae, whereas Sclerotinia sclerotiorum and Botrytis cinerea are cosmopolitan broad host-range plant pathogens, Sclerotinia borealis has a psychrophilic lifestyle with a low optimal growth temperature, a narrow host range and geographic distribution. To spread successfully, S. borealis must synthesize proteins adapted to function in its specific environment. The search for signatures of adaptation to S. borealis lifestyle may therefore help revealing proteins critical for colonization of the environment by Sclerotiniaceae fungi. Here, we analyzed amino acids usage and intrinsic protein disorder in alignments of groups of orthologous proteins from the three Sclerotiniaceae species. We found that enrichment in Thr, depletion in Glu and Lys, and low disorder frequency in hot loops are significantly associated with S. borealis proteins. We designed an index to report bias in these properties and found that high index proteins were enriched among secreted proteins in the three Sclerotiniaceae fungi. High index proteins were also enriched in function associated with plant colonization in S. borealis, and in in planta-induced genes in S. sclerotiorum. We highlight a novel putative antifreeze protein and a novel putative lytic polysaccharide monooxygenase identified through our pipeline as candidate proteins involved in colonization of the environment. Our findings suggest that similar protein signatures associate with S. borealis lifestyle and with secretion in the Sclerotiniaceae. These signatures may be useful for identifying proteins of interest as targets for the management of plant diseases.

  20. Molecular modeling and structural characterization of a high glycine-tyrosine hair keratin associated protein. (United States)

    Singh, Rakesh S; Palmer, Jeremy C; Pudney, Paul D A; Paul, Prem K C; Johannessen, Christian; Debenedetti, Pablo G; Raut, Janhavi; Lee, Ken; Noro, Massimo; Tiemessen, David


    High glycine-tyrosine (HGT) proteins are an important constituent of the keratin associated proteins (KAPs) present in human hair. The glassy state physics of hair fibres are thought to be largely regulated by KAPs, which exist in an amorphous state and are readily affected by environmental conditions. However, there are no studies characterizing the individual KAPs. In this paper, we present the first step to fill this gap by computational modeling and experimental studies on a HGT protein, KAP8.1. In particular, we have modeled the three-dimensional structure of this 63-residue protein using homology information from an anti-freeze protein in snow flea. The model for KAP8.1 is characterized by four strands of poly-proline II (or PPII) type helical secondary structures, held together by two cysteine disulphide bridges. Computer simulations confirm the stability of the modelled structure and show that the protein largely samples the PPII and β-sheet conformations during the molecular dynamics simulations. Spectroscopic studies including Raman, IR and vibrational circular dichroism have also been performed on synthesized KAP8.1. The experimental studies suggest that KAP8.1 is characterised by β-sheet and PPII structures, largely consistent with the simulation studies. The model built in this work is a good starting point for further simulations to study in greater depth the glassy state physics of hair, including its water sorption isotherms, glass transition, and the effect of HGT proteins on KAP matrix plasticization. These results are a significant step towards our goal of understanding how the properties of hair can be affected and manipulated under different environmental conditions of temperature, humidity, ageing and small molecule additives.

  1. Kinetics of the Methanol Reaction with OH at Interstellar, Atmospheric, and Combustion Temperatures. (United States)

    Gao, Lu Gem; Zheng, Jingjing; Fernández-Ramos, Antonio; Truhlar, Donald G; Xu, Xuefei


    The OH radical is the most important radical in combustion and in the atmosphere, and methanol is a fuel and antifreeze additive, model biofuel, and trace atmospheric constituent. These reagents are also present in interstellar space. Here we calculate the rate constants, branching ratios, and kinetic isotope effects (KIEs) of the hydrogen abstraction reaction of methanol by OH radical in a broad temperature range of 30-2000 K, covering interstellar space, the atmosphere, and combustion by using the competitive canonical unified statistical (CCUS) model in both the low-pressure and high-pressure limits and, for comparison, the pre-equilibrium model. Coupled cluster CCSD(T)-F12a theory and multi-reference CASPT2 theory were used to carry out benchmark calculations of the stationary points on the potential energy surface to select the most appropriate density functional method for direct dynamics calculations of rate constants. We find a significant effect of the anharmonicity of high-frequency modes of transition states on the low-temperature rate constant, and we show how tunneling leads to an unusual negative temperature dependence of the rate constants in the range 200 K > T > 100 K. The calculations also demonstrate the importance of the extent of stabilization of the pre-reactive complex. The capture rate for the formation of the complex is the dominant dynamical bottleneck for T < 100 K, and it leads to weak temperature dependence of the rate below 100 K in the high-pressure-limit of the CCUS model. We also report the pressure dependence of branching ratios (which are hard to measure so theory is essential) and the KIEs, and we report an unusual nonmonotonic variation of the KIE in the high-pressure limit at low temperatures.

  2. Studies on the new fuels with Santilli magnecular structure and their industrial applications

    Energy Technology Data Exchange (ETDEWEB)

    Pandhurnekar, Chandrashekhar P., E-mail: [Shri Ramdeobaba College of Engineering and Management, Nagpur, Maharashtra 440 013 (India)


    Professor R. M. Santilli, the Italian-American physicist, for the first time in the history of Science, presented the theoretical and experimental evidence on the existence of the new chemical species of “magnecules” [1]. This new species mainly consist of individual atoms, radicals and conventional molecules bonded together with stable clusters under the new attractive force primarily originating from torroidal polarization of orbitals of atomic electrons under strong magnetic field. The main contribution in this area was the production of Magnegas{sup TM}, new clean fuels developed by Prof. Santilli, which are produced as byproducts of recycling nonradioactive liquid feedstock such as antifreeze waste, engine oil waste, town sewage, crude oil, etc., and generally vary with the liquid used for their production. A new technology, called Plasma Arc FlowTM, flows the waste through a submerged electric arc between conventional electrodes. The arc decomposes the liquid molecules into their atomic constituents, and forms a plasma in the immediate vicinity of the electrodes at about 10,000{sup 0} F. The technology then moves the plasma away from the electrodes, and controls its recombination into environmentally acceptable fuels. In fact, the exhaust of magnegases shows: absence of carcinogenic or other toxic substances; breathable oxygen up 14 percent; and carbon dioxide down to 0.01 percent. Since, in addition, the new fuels can be produced everywhere, and have environmentally acceptable exhausts, Magnegases offer promising possibilities to satisfy our ever increasing energy needs, as well as to contain the alarming environmental problems caused by fossil fuels. Thus, it was thought worthwhile to present some of the industrial applications of environmentally benign fuel consisting magnecular bonds [2, 3, 4, 5]. Also in the present communications, some of the experimental evidences of Santilli’s new chemical species i. e. Magnecules which had been published

  3. Cryopreservation of boar sperm induces differential microRNAs expression. (United States)

    Zhang, Yan; Dai, Dinghui; Chang, Yu; Li, Yuan; Zhang, Ming; Zhou, Guangbin; Peng, Zhanghua; Zeng, Changjun


    Lower conception rates and litter sizes limit the wide use of artificial insemination with frozen-thawed boar sperm, due to a lack of understanding of the mechanisms that cause cryodamage and cryoinjury to sperm during cryopreservation. CryoMiRs, a family of freeze-related microRNAs (miRNAs), are associated with freeze tolerance, and regulate metabolism in mammalian hibernators and insects. Thus, we speculate that miRNAs maybe involved in the regulation of the freeze-thaw process and may affect boar sperm function. In this study, we studied the differential expression of 46 miRNAs that have roles in spermatogenesis, sperm maturation, and sperm quality in response to cryopreservation (with or without 3% glycerol). The results indicated that, in response to cryopreservation with 3% glycerol, 14 miRNAs were significantly up-regulated, but only two miRNAs (miR-22 and miR-450b-5p) were significantly down-regulated, relative to fresh sperm. Preservation with 3% glycerol caused up-regulation of 17 miRNAs, but only caused down-regulation of one miRNA (miR-24), relative to sperm cryopreserved without glycerol. Functional annotations of these differentially expressed miRNAs indicated that these miRNAs and their targets are mainly associated with metabolic and cellular processes. Therefore, our findings show that cryopreservation results in changes in miRNA expression, and suggest that the anti-freeze mechanisms of boar sperm need to be studied further. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Genome reconstructions indicate the partitioning of ecological functions inside a phytoplankton bloom in the Amundsen Sea, Antarctica

    Directory of Open Access Journals (Sweden)

    Tom O Delmont


    Full Text Available Antarctica polynyas support intense phytoplankton blooms, impacting their environment by a substantial depletion of inorganic carbon and nutrients. These blooms are dominated by the colony-forming haptophyte Phaeocystis antarctica and they are accompanied by a distinct bacterial population. Yet, the ecological role these bacteria may play in P. antarctica blooms awaits elucidation of their functional gene pool and of the geochemical activities they support. Here, we report on a metagenome (῀160 million reads analysis of the microbial community associated with a P. antarctica bloom event in the Amundsen Sea polynya (West Antarctica. Genomes of the most abundant Bacteroidetes and Proteobacteria populations have been reconstructed and a network analysis indicates a strong functional partitioning of these bacterial taxa. Three of them (SAR92, and members of the Oceanospirillaceae and Cryomorphaceae are found in close association with P. antarctica colonies. Distinct features of their carbohydrate, nitrogen, sulfur and iron metabolisms may serve to support mutualistic relationships with P. antarctica. The SAR92 genome indicates a specialization in the degradation of fatty acids and dimethylsulfoniopropionate (compounds released by P. antarctica into dimethyl sulfide, an aerosol precursor. The Oceanospirillaceae genome carries genes that may enhance algal physiology (cobalamin synthesis. Finally, the Cryomorphaceae genome is enriched in genes that function in cell or colony invasion. A novel pico-eukaryote, Micromonas related genome (19.6 Mb, ~94% completion was also recovered. It contains the gene for an anti-freeze protein, which is lacking in Micromonas at lower latitudes. These draft genomes are representative for abundant microbial taxa across the Southern Ocean surface.

  5. A rare polyglycine type II-like helix motif in naturally occurring proteins. (United States)

    Warkentin, Eberhard; Weidenweber, Sina; Schühle, Karola; Demmer, Ulrike; Heider, Johann; Ermler, Ulrich


    Common structural elements in proteins such as α-helices or β-sheets are characterized by uniformly repeating, energetically favorable main chain conformations which additionally exhibit a completely saturated hydrogen-bonding network of the main chain NH and CO groups. Although polyproline or polyglycine type II helices (PP II or PG II ) are frequently found in proteins, they are not considered as equivalent secondary structure elements because they do not form a similar self-contained hydrogen-bonding network of the main chain atoms. In this context our finding of an unusual motif of glycine-rich PG II -like helices in the structure of the acetophenone carboxylase core complex is of relevance. These PG II -like helices form hexagonal bundles which appear to fulfill the criterion of a (largely) saturated hydrogen-bonding network of the main-chain groups and therefore may be regarded in this sense as a new secondary structure element. It consists of a central PG II -like helix surrounded by six nearly parallel PG II -like helices in a hexagonal array, plus an additional PG II -like helix extending the array outwards. Very related structural elements have previously been found in synthetic polyglycine fibers. In both cases, all main chain NH and CO groups of the central PG II -helix are saturated by either intra- or intermolecular hydrogen-bonds, resulting in a self-contained hydrogen-bonding network. Similar, but incomplete PG II -helix patterns were also previously identified in a GTP-binding protein and an antifreeze protein. © 2017 Wiley Periodicals, Inc.

  6. Infection by chikungunya virus modulates the expression of several proteins in Aedes aegypti salivary glands

    Directory of Open Access Journals (Sweden)

    Tchankouo-Nguetcheu Stephane


    Full Text Available Abstract Background Arthropod-borne viral infections cause several emerging and resurging infectious diseases. Among the diseases caused by arboviruses, chikungunya is responsible for a high level of severe human disease worldwide. The salivary glands of mosquitoes are the last barrier before pathogen transmission. Methods We undertook a proteomic approach to characterize the key virus/vector interactions and host protein modifications that occur in the salivary glands that could be responsible for viral transmission by using quantitative two-dimensional electrophoresis. Results We defined the protein modulations in the salivary glands of Aedes aegypti that were triggered 3 and 5 days after an oral infection (3 and 5 DPI with chikungunya virus (CHIKV. Gel profile comparisons showed that CHIKV at 3 DPI modulated the level of 13 proteins, and at 5 DPI 20 proteins. The amount of 10 putatively secreted proteins was regulated at both time points. These proteins were implicated in blood-feeding or in immunity, but many have no known function. CHIKV also modulated the quantity of proteins involved in several metabolic pathways and in cell signalling. Conclusion Our study constitutes the first analysis of the protein response of Aedes aegypti salivary glands infected with CHIKV. We found that the differentially regulated proteins in response to viral infection include structural proteins and enzymes for several metabolic pathways. Some may favour virus survival, replication and transmission, suggesting a subversion of the insect cell metabolism by arboviruses. For example, proteins involved in blood-feeding such as the short D7, an adenosine deaminase and inosine-uridine preferring nucleoside hydrolase, may favour virus transmission by exerting an increased anti-inflammatory effect. This would allow the vector to bite without the bite being detected. Other proteins, like the anti-freeze protein, may support vector protection.

  7. Fish from the Southern Ocean: biodiversity, ecology and conservation challenges

    Directory of Open Access Journals (Sweden)

    Marino Vacchi


    Living and functioning at subzero temperatures implied important adaptations, including freezing avoidance by antifreeze glycoproteins ( AFGPs. Among the system-wide adaptive traits holding major ecological implications, the acquisition of secondary pelagicism in some species (plesiomorphically devoid of swim-bladder is a major. In those notothenioids, lipid deposition and reduced ossification allowed to achieve partial or full neutral buoyancy, and enabled expansion into semi-pelagic, pelagic, and cryopelagic habitats. Such an impressive ecological expansion has allowed several notothenioids to play a primary role in the Antarctic marine ecosystems. On the other side, their fine adaptation to the environment, might expose these fishes to risks that need to be properly considered and addressed. For instance, a relationship between the Antarctic silverfish (Pleuragramma antarctica, a key species in the coastal Antarctic ecosystem and the sea-ice, has recently been assessed, thus making this species potentially threatened by the ongoing climatic change, with implications for the whole ecosystem. In addition, some Antarctic fish, such as toothfishes (Dissostichus eleginoides and Dissostichus mawsoni are primary targets of industrial fish harvesting in the SO. To increase and update the scientific knowledge on these species is mandatory in order to improve the management of Antarctic marine resources, in response to the increasing international request of exploitation. This task is presently being conducted by CCAMLR (Commission for the Conservation of Antarctic Marine Living Resources, along with fighting the illegal, unreported and unregulated (IUU fishing and with the establishment of MPAs (Marine Protected Areas in various sectors of the Southern Ocean.

  8. Ethylene glycol toxicity presenting with non-anion gap metabolic acidosis. (United States)

    Soghoian, Sari; Sinert, Richard; Wiener, Sage W; Hoffman, Robert S


    Ethylene glycol classically produces an elevated anion gap metabolic acidosis. We report a series of patients with ethylene glycol toxicity with a component of non-anion gap metabolic acidosis without known associated confounding factors. A retrospective review of Poison Control Center records were searched more than 8 years (2000-2007) for ethylene glycol and antifreeze. Cases were reviewed and excluded for miscoding, information calls, animal exposures, or non-ingestion exposures. The bicarbonate gap, or delta ratio (DR), was calculated using the formula: DR = (AG - 12)/[24 - measured serum where anion gap (AG) = [Na(+)] - [Cl(-)] - , all in mEq/l. Non-anion gap metabolic acidosis was considered present when the DR anion gap metabolic acidosis at presentation. Their calculated anion gap was 14-28, and measured serum ranged from 2-20 mEq/l. A normal anion gap was present in two patients who presented with non-anion gap metabolic acidosis. The DR ranged from 0.28-0.95. Seven out of 14 patients with non-anion gap metabolic acidosis had elevated serum [Cl(-)]. In the other cases, no explanation for the non-anion gap metabolic acidosis could be determined. The absence of a significant anion gap elevation in the setting of metabolic acidosis after ethylene glycol ingestion without other confounding factors (such as ethanol, lithium carbonate or bromide) has not previously been recognized. Clinicians should be aware of the potential for non-anion gap metabolic acidosis in patients with ethylene glycol toxicity, and should not exclude the diagnosis in patients who present with a non-anion gap metabolic acidosis. Further study is needed to determine the mechanisms by which this occurs.

  9. Toxicology and carcinogenesis studies of dipropylene glycol in rats and mice. (United States)

    Hooth, Michelle J; Herbert, Ronald A; Haseman, Joseph K; Orzech, Denise P; Johnson, Jerry D; Bucher, John R


    Dipropylene glycol (DPG) is a component of many commercial products such as antifreeze, air fresheners, cosmetic products, solvents, and plastics. Male and female F344/N rats and B6C3F1 mice were exposed to DPG in the drinking water for 2 weeks, 3 months, or 2 years. In the 2-week and 3-month studies, rats and mice were exposed to 0, 5000, 10,000, 20,000, 40,000, or 80,000 ppm DPG. There was no mortality in the 2-week studies. In the 3-month rat study, all animals survived to the end of the study. Liver weights of rats exposed to 10,000 ppm or greater and kidney weights of rats exposed to 40,000 and 80,000 ppm were greater than those of the controls. The incidences of liver and kidney lesions were significantly increased in males exposed to 20,000 ppm or greater and females exposed to 80,000 ppm. Focal olfactory epithelial degeneration was present in all rats exposed to 80,000 ppm. In males, the incidences of testicular atrophy, epididymal hypospermia, and preputial gland atrophy were significantly increased in the 80,000 ppm group. In the 3-month mouse study, three males and one female exposed to 80,000 ppm died. Liver weights were increased, as was the incidence of centrilobular hypertrophy in males exposed to 40,000 ppm and males and females exposed to 80,000 ppm. In the 2-year studies, exposure groups were 0, 2500 (rats only), 10,000, 20,000 (mice only) or 40,000 ppm DPG. Survival of male rats exposed to 40,000 ppm and mean body weights of males and females exposed to 40,000 ppm were significantly less than controls. In male rats, exposure to DPG resulted in increased incidences and severities of nephropathy and secondary lesions in the parathyroid and forestomach. Increased incidences of focal histiocytic and focal granulomatous inflammation of the liver were also observed. In male and female rats, there were increased incidences of bile duct hyperplasia and changes in the olfactory epithelium of the nose. In mice, survival of males and females was similar to

  10. Suppression of Frost Nucleation Achieved Using the Nanoengineered Integral Humidity Sink Effect. (United States)

    Sun, Xiaoda; Rykaczewski, Konrad


    coatings can be designed to combine optimal antifrosting functionality with a superhydrophobic water repelling exterior to provide coatings that can robustly prevent frost, rime, and glaze accumulation. By minimizing the required amount of antifreeze, this anti-icing method can have minimal operational cost and environmental impact.

  11. Deletion of meso-2,3-butanediol dehydrogenase gene budC for enhanced D-2,3-butanediol production in Bacillus licheniformis (United States)


    Background D-2,3-butanediol has many industrial applications such as chiral reagents, solvents, anti-freeze agents, and low freezing point fuels. Traditional D-2,3-butanediol producing microorganisms, such as Klebsiella pneumonia and K. xoytoca, are pathogenic and not capable of producing D-2,3-butanediol at high optical purity. Bacillus licheniformis is a potential 2,3-butanediol producer but the wild type strain (WX-02) produces a mix of D- and meso-type isomers. BudC in B. licheniformis is annotated as 2,3-butanediol dehydrogenase or acetoin reductase, but no pervious experiment was performed to verify this hypothesis. Results We developed a genetically modified strain of B. licheniformis (WX-02 ΔbudC) as a D-2,3-butanediol producer with high optimal purity. A marker-less gene deletion protocol based on a temperature sensitive knock-out plasmid T2-Ori was used to knock out the budC gene in B. licheniformis WX-02. The budC knock-out strain successfully abolished meso-2,3-butanediol production with enhanced D-2,3-butanediol production. No meso-BDH activity was detectable in cells of this strain. On the other hand, the complementary strain restored the characteristics of wild strain, and produced meso-2,3-butanediol and possessed meso-BDH activity. All of these data suggested that budC encoded the major meso-BDH catalyzing the reversible reaction from acetoin to meso-2,3-butanediol in B. licheniformis. The budC knock-out strain produced D-2,3-butanediol isomer only with a high yield of 30.76 g/L and a productivity of 1.28 g/L-h. Conclusions We confirmed the hypothesis that budC gene is responsible to reversibly transfer acetoin to meso-2,3-butanediol in B. licheniformis. A mutant strain of B. licheniformis with depleted budC gene was successfully developed and produced high level of the D-2,3-butanediol with high optimal purity. PMID:24475980

  12. Phase behaviour and thermoelastic properties of ammonia hydrate and ice polymorphs from 0 - 2 GPa (United States)

    Fortes, A. D.; Wood, I. G.; Vocadlo, L.


    Ammonia remains amongst the most plausible planetary "antifreeze" agents, and its physical properties in hydrate compounds under the appropriate conditions (roughly 0 - 5 GPa, 100 - 300 K) must be known in order for it to be accommodated in planetary models. The pressure melting curve, and the expected polymorphism of the stoichiometric ammonia hydrates have implications for the internal structure of large icy moons like Titan, leading to phase layering and the possible persistence of deep subsurface oceans, the latter being sites of high astrobiological potential. Aqueous ammonia is also a candidate substance involved in cryomagmatism on Titan, and again the melting behaviour, and densities of liquids and solids, in the ammonia-water system must be known to model properly the partial melting and propagation of magma. We describe the results of a series of powder neutron diffraction experiments over the range 0 - 2.0 GPa, 150 - 280 K which were carried out with the objective of determining the phase behaviour and thermoelastic properties of ammonia dihydrate. In addition to the low-pressure cubic crystalline phase, ADH I, we have identified two closely related monoclinic polymorphs of ammonia dihydrate (ADH IIa and IIb) in the range 0.45 - 0.60 GPa (at 175 K), and have determined that this phase dissociates to a mixture of ammonia monohydrate phase II and ice II when warmed to ~190 K, which in turn melts at a binary eutectic at ~196 K; AMH II has a large (Z = 16) orthorhombic unit cell. Above 0.60 GPa, an orthorhombic polymorph of ammonia dihydrate, which we have referred to previously as ADH IV, persists to pressures > 3 GPa, and appears to be the liquidus phase over this whole pressure range. We have observed this phase co- existing with both ice II and ice VI. Here we describe the most plausible synthesis of the high-pressure phase diagram which explains our observations, and provide measurements of the densities, thermal expansion, bulk moduli, and crystal

  13. Hydrothermal Habitats: Measurements of Bulk Microbial Elemental Composition, and Models of Hydrothermal Influences on the Evolution of Dwarf Planets (United States)

    Neveu, Marc Francois Laurent

    Finding habitable worlds is a key driver of solar system exploration. Many solar system missions seek environments providing liquid water, energy, and nutrients, the three ingredients necessary to sustain life. Such environments include hydrothermal systems, spatially-confined systems where hot aqueous fluid circulates through rock by convection. I sought to characterize hydrothermal microbial communities, collected in hot spring sediments and mats at Yellowstone National Park, USA, by measuring their bulk elemental composition. To do so, one must minimize the contribution of non-biological material to the samples analyzed. I demonstrate that this can be achieved using a separation method that takes advantage of the density contrast between cells and sediment and preserves cellular elemental contents. Using this method, I show that in spite of the tremendous physical, chemical, and taxonomic diversity of Yellowstone hot springs, the composition of microorganisms there is surprisingly ordinary. This suggests the existence of a stoichiometric envelope common to all life as we know it. Thus, future planetary investigations could use elemental fingerprints to assess the astrobiological potential of hydrothermal settings beyond Earth. Indeed, hydrothermal activity may be widespread in the solar system. Most solar system worlds larger than 200 km in radius are dwarf planets, likely composed of an icy, cometary mantle surrounding a rocky, chondritic core. I enhance a dwarf planet evolution code, including the effects of core fracturing and hydrothermal circulation, to demonstrate that dwarf planets likely have undergone extensive water-rock interaction. This supports observations of aqueous products on their surfaces. I simulate the alteration of chondritic rock by pure water or cometary fluid to show that aqueous alteration feeds back on geophysical evolution: it modifies the fluid antifreeze content, affecting its persistence over geological timescales; and the

  14. Silicon Utilizing Microbial Bioactivities in the Biosphere (United States)

    Sen, M. M.; Das, S.


    Diatoms are unicellular eukaryotic algae and an important member of the silicon utilizing organisms, that generate ~20% of the ~100 billion metric tons of organic carbon produced through photosynthesis on Earth each year. Fragilariopsis is a dominating psychrophilic diatom genus in polar sea ice. The two species Fragilariopsis cylindrus and Fragilariopsis curta are able to grow and divide below freezing temperature. Antifreeze proteins (AFPs), involved in cold adaptation in several psychrophilic organisms, are widespread in this two polar species. Achanthes minutissima isolated as dominant diatom has degradable effects involving petroleum hydocarbons. Phaeodactylum tricornutum, have antibacterial activity and the fatty acid, eicosapentaenoic acid (EPA), has been identified as one compound responsible for this activity. Other antibacterial compounds are monounsaturated fatty acid (9Z)-hexadecenoic acid (palmitoleic acid; C16:1 n-7) and the relatively unusual polyunsaturated fatty acid (6Z, 9Z, 12Z)-hexadecatrienoic acid (HTA; C16:3 n-4). Both are active against Gram-positive bacteria and many Gram-negative pathogen. Palmitoleic acid is active at micro-molar concentrations, kills bacteria rapidly, and is highly active against multidrug-resistant Staphylococcus aureus. Domoic acid -a neurotoxin produced by Pseudo-nitzschia accumulates in marine invertebrates. Evidences of sea lion (Zalophus californianus) and human poisoning following consumption of contaminated blue mussels (Mytilus edulis) is mainly due to this toxin. Among the most prominent features described in human beings was memory impairment which led to the name Amnesic Shellfish Poisoning [ASP]. Silicon utilizing organisms can act as a bioindicator of environmental contamination, thus a rapid change in phytochelatins to both the increase in and the withdrawal of environmental Cd stress was found in Thalassiosira nordenskioeldii. Some of them also can produce biofuels particularly diatoms have significant

  15. Modeling Late-State Serpentinization on Enceladus and Implications for Methane-Utilizing Microbial Metabolisms (United States)

    Hart, R.; Cardace, D.


    Modeling investigations of Enceladus and other icy-satellites have included physicochemical properties (Sohl et al., 2010; Glein et al., 2015; Neveu et al., 2015), geophysical prospects of serpentinization (Malamud and Prialnik, 2016; Vance et al., 2016), and aqueous geochemistry across different antifreeze fluid-rock scenarios (Neveu et al., 2017). To more effectively evaluate the habitability of Enceladus, in the context of recent observations (Waite et al., 2017), we model the potential bioenergetic pathways that would be thermodynamically favorable at the interface of hydrothermal water-rock reactions resulting from late stage serpentinization (>90% serpentinized), hypothesized on Enceladus. Building on previous geochemical model outputs of Enceladus (Neveu et al., 2017), and bioenergetic modeling (as in Amend and Shock, 2001; Cardace et al., 2015), we present a model of late stage serpentinization possible at the water-rock interface of Enceladus, and report changing activities of chemical species related to methane utilization by microbes over the course of serpentinization using the Geochemist's Workbench REACT code [modified Extended Debye-Hückel (Helgeson, 1969) using the thermodynamic database of SUPCRT92 (Johnson et al., 1992)]. Using a model protolith speculated to exist at Enceladus's water-rock boundary, constrained by extraterrestrial analog analytical data for subsurface serpentinites of the Coast Range Ophiolite (Lower Lake, CA, USA) mélange rocks, we deduce evolving habitability conditions as the model protolith reacts with feasible, though hypothetical, planetary ocean chemistries (from Glien et al., 2015, and Neveu et al., 2017). Major components of modeled oceans, Na-Cl, Mg-Cl, and Ca-Cl, show shifts in the feasibility of CO2-CH4-H2 driven microbial habitability, occurring early in the reaction progress, with methanogenesis being bioenergetically favored. Methanotrophy was favored late in the reaction progress of some Na-Cl systems and in the

  16. The indoor volatile organic compound (VOC) characteristics and source identification in a new university campus in Tianjin, China. (United States)

    Kang, Jian; Liu, Junjie; Pei, Jingjing


    This study investigates the volatile organic compounds (VOCs) constituents and concentration levels on a new university campus, where all of the buildings including classrooms and student dormitories were newly built and decorated within 1 year. Investigated indoor environments include dormitories, classrooms, and the library. About 30 dormitory buildings with different furniture loading ratios were measured. The characteristics of the indoor VOCs species are analyzed and possible sources are identified. The VOCs were analyzed with gas chromatography-mass spectroscopy (GC-MS). It was found that the average total VOC (TVOC) concentration can reach 2.44 mg/m 3 . Alkenes were the most abundant VOCs in dormitory rooms, contributing up to 86.5% of the total VOCs concentration. The concentration of α-pinene is the highest among the alkenes. Unlike the dormitory rooms, there is almost no room with TVOC concentration above 0.6 mg/m 3 in classroom and library buildings. Formaldehyde concentration in the dormitory rooms increased about 23.7% after the installation of furniture, and the highest level reached 0.068 mg/m 3 . Ammonia released from the building antifreeze material results in an average indoor concentration of 0.28 mg/m 3 , which is 100% over the threshold and should be seriously considered. Further experiments were conducted to analyze the source of the α-pinene and some alkanes in dormitory rooms. The results showed that the α-pinene mainly comes from the bed boards, while the wardrobes are the main sources of alkanes. The contribution of the pinewood bed boards to the α-pinene and TVOC concentration can reach up to above 90%. The same type rooms were sampled 1 year later and the decay rate of α-pinene is quite high, close to 100%, so that it almost cannot be detected in the sampled rooms. Analysis of indoor volatile organic compounds (VOCs) in newly built campus buildings in China identified the specific constituents of indoor VOCs contaminants exposed to

  17. NTP-CERHR expert panel report on the reproductive and developmental toxicity of methanol. (United States)

    Shelby, Michael; Portier, Christopher; Goldman, Lynn; Moore, John; Iannucci, Annette; Jahnke, Gloria; Donkin, Steve


    The National Toxicology Program (NTP) and the National Institute of Environmental Health Sciences (NIEHS) established the NTP Center for the Evaluation of Risks to Human Reproduction (CERHR) in June 1998. The purpose of the Center is to provide timely, unbiased, scientifically sound evaluations of human and experimental evidence for adverse effects on reproduction, including development, caused by agents to which humans may be exposed. Methanol was selected for evaluation by the CERHR based on high production volume, extent of human exposure, and published evidence of reproductive or developmental toxicity. Methanol is used in chemical syntheses and as an industrial solvent. It is a natural component of the human diet and is found in consumer products such as paints, antifreeze, cleaning solutions, and adhesives. It is used in race car fuels and there is potential for expanded use as an automobile fuel. This evaluation is the result of a 10-month effort by a 12-member panel of government and non-government scientists that culminated in a public Expert Panel meeting. This report has been reviewed by CERHR staff scientists, and by members of the Methanol Expert Panel. Copies have been provided to the CERHR Core Committee, which is made up of representatives of NTP-participating agencies. This report is a product of the Expert Panel and is intended to (1). interpret the strength of scientific evidence that a given exposure or exposure circumstance may pose a hazard to reproduction and the health and welfare of children; (2). provide objective and scientifically thorough assessments of the scientific evidence that adverse reproductive/development health effects are associated with exposure to specific chemicals or classes of chemicals, including descriptions of any uncertainties that would diminish confidence in assessment of risks; and (3). identify knowledge gaps to help establish research and testing priorities. The expert panel report becomes a central part of the

  18. Wintertime Emissions from Produced Water Ponds (United States)

    Evans, J.; Lyman, S.; Mansfield, M. L.


    surfaces. These compounds are highly reactive and, because of their relatively high water solubility, tend to concentrate in produced water. The average methanol emission rate from unfrozen pond surfaces was more than 100 mg m-2 h-1. Methanol, used as an antifreeze and anti-scaler in the oil and gas industry, is abundant during winter inversions in the Uintah Basin and may also be a significant precursor to ozone production. Total VOC and methanol emissions from produced water ponds during winter were estimated to be 178 and 83 tons month-1, respectively, for the entire Uintah Basin.

  19. Evolution of Planetary Ice-Ocean Systems: Effects of Salinity (United States)

    Allu Peddinti, D.; McNamara, A. K.


    Planetary oceanography is enjoying renewed attention thanks to not only the detection of several exoplanetary ocean worlds but also due to the expanding family of ocean worlds within our own star system. Our solar system is now believed to host about nine ocean worlds including Earth, some dwarf planets and few moons of Jupiter and Saturn. Amongst them, Europa, like Earth is thought to have an ice Ih-liquid water system. However, the thickness of the Europan ice-ocean system is much larger than that of the Earth. The evolution of this system would determine the individual thicknesses of the ice shell and the ocean. In turn, these thicknesses can alter the course of evolution of the system. In a pure H2O system, the thickness of the ice shell would govern if heat loss occurs entirely by conduction or if the shell begins to convect as it attains a threshold thickness. This switch between conduction-convection regimes could determine the longevity of the subsurface ocean and hence define the astrobiological potential of the planetary body at any given time. In reality, however, the system is not pure water ice. The detected induced magnetic field infers a saline ocean layer. Salts are expected to act as an anti-freeze allowing a subsurface ocean to persist over long periods but the amount of salts would determine the extent of that effect. In our current study, we use geodynamic models to examine the effect of salinity on the evolution of ice-ocean system. An initial ocean with different salinities is allowed to evolve. The effect of salinity on thickness of the two layers at any time is examined. We also track how salinity controls the switch between conductive-convective modes. The study shows that for a given time period, larger salinities can maintain a thick vigorously convecting ocean while the smaller salinities behave similar to a pure H2O system leading to a thick convecting ice-shell. A range of salinities identified can potentially predict the current state

  20. Oil palm EgCBF3 conferred stress tolerance in transgenic tomato plants through modulation of the ethylene signaling pathway. (United States)

    Ebrahimi, Mortaza; Abdullah, Siti Nor Akmar; Abdul Aziz, Maheran; Namasivayam, Parameswari


    CBF/DREB1 is a group of transcription factors that are mainly involved in abiotic stress tolerance in plants. They belong to the AP2/ERF superfamily of plant-specific transcription factors. A gene encoding a new member of this group was isolated from ripening oil palm fruit and designated as EgCBF3. The oil palm fruit demonstrates the characteristics of a climacteric fruit like tomato, in which ethylene has a major impact on the ripening process. A transgenic approach was used for functional characterization of the EgCBF3, using tomato as the model plant. The effects of ectopic expression of EgCBF3 were analyzed based on expression profiling of the ethylene biosynthesis-related genes, anti-freeze proteins (AFPs), abiotic stress tolerance and plant growth and development. The EgCBF3 tomatoes demonstrated altered phenotypes compared to the wild type tomatoes. Delayed leaf senescence and flowering, increased chlorophyll content and abnormal flowering were the consequences of overexpression of EgCBF3 in the transgenic tomatoes. The EgCBF3 tomatoes demonstrated enhanced abiotic stress tolerance under in vitro conditions. Further, transcript levels of ethylene biosynthesis-related genes, including three SlACSs and two SlACOs, were altered in the transgenic plants' leaves and roots compared to that in the wild type tomato plant. Among the eight AFPs studied in the wounded leaves of the EgCBF3 tomato plants, transcript levels of SlOSM-L, SlNP24, SlPR5L and SlTSRF1 decreased, while expression of the other four, SlCHI3, SlPR1, SlPR-P2 and SlLAP2, were up-regulated. These findings indicate the possible functions of EgCBF3 in plant growth and development as a regulator of ethylene biosynthesis-related and AFP genes, and as a stimulator of abiotic stress tolerance. Copyright © 2016 Elsevier GmbH. All rights reserved.

  1. Fluid Physics and Macromolecular Crystal Growth in Microgravity (United States)

    Helliwell, John R.; Snell, Edward H.; Chayen, Naomi E.; Judge, Russell A.; Boggon, Titus J.; Pusey, M. L.; Rose, M. Franklin (Technical Monitor)


    The first protein crystallization experiment in microgravity was launched in April, 1981 and used Germany's Technologische Experimente unter Schwerelosigkeit (TEXUS 3) sounding rocket. The protein P-galactosidase (molecular weight 465Kda) was chosen as the sample with a liquid-liquid diffusion growth method. A sliding device brought the protein, buffer and salt solution into contact when microgravity was reached. The sounding rocket gave six minutes of microgravity time with a cine camera and schlieren optics used to monitor the experiment, a single growth cell. In microgravity a strictly laminar diffusion process was observed in contrast to the turbulent convection seen on the ground. Several single crystals, approx 100micron in length, were formed in the flight which were of inferior but of comparable visual quality to those grown on the ground over several days. A second experiment using the same protocol but with solutions cooled to -8C (kept liquid with glycerol antifreeze) again showed laminar diffusion. The science of macromolecular structural crystallography involves crystallization of the macromolecule followed by use of the crystal for X-ray diffraction experiments to determine the three dimensional structure of the macromolecule. Neutron protein crystallography is employed for elucidation of H/D exchange and for improved definition of the bound solvent (D20). The structural information enables an understanding of how the molecule functions with important potential for rational drug design, improved efficiency of industrial enzymes and agricultural chemical development. The removal of turbulent convection and sedimentation in microgravity, and the assumption that higher quality crystals will be produced, has given rise to the growing number of crystallization experiments now flown. Many experiments can be flown in a small volume with simple, largely automated, equipment - an ideal combination for a microgravity experiment. The term "protein crystal growth

  2. Production of (2R, 3R)-2,3-butanediol using engineeredPichia pastoris: strain construction, characterization and fermentation. (United States)

    Yang, Zhiliang; Zhang, Zisheng


    2,3-butanediol (2,3-BD) is a bulk platform chemical with various potential applications such as aviation fuel. 2,3-BD has three optical isomers: (2R, 3R)-, (2S, 3S)- and meso-2,3-BD. Optically pure 2,3-BD is a crucial precursor for the chiral synthesis and it can also be used as anti-freeze agent due to its low freezing point. 2,3-BD has been produced in both native and non-native hosts. Several pathogenic bacteria were reported to produce 2,3-BD in mixture of its optical isomers including Klebsiella pneumoniae and Klebsiella oxytoca . Engineered hosts based on episomal plasmid expression such as Escherichia coli , Saccharomyces cerevisiae and Bacillus subtilis are not ideal for industrial fermentation due to plasmid instability. Pichia pastoris is generally regarded as safe and a well-established host for high-level heterologous protein production. To produce pure (2R, 3R)-2,3-BD enantiomer, we developed a P. pastoris strain by introducing a synthetic pathway. The als S and als D genes from B. subtilis were codon-optimized and synthesized. The BDH1 gene from S. cerevisiae was cloned. These three pathway genes were integrated into the genome of P. pastoris and expressed under the control of GAP promoter. Production of (2R, 3R)-2,3-BD was achieved using glucose as feedstock. The optical purity of (2R, 3R)-2,3-BD was more than 99%. The titer of (2R, 3R)-2,3-BD reached 12 g/L with 40 g/L glucose as carbon source in shake flask fermentation. The fermentation conditions including pH, agitation speeds and aeration rates were optimized in batch cultivations. The highest titer of (2R, 3R)-2,3-BD achieved in fed-batch fermentation using YPD media was 45 g/L. The titer of 2,3-BD was enhanced to 74.5 g/L through statistical medium optimization. The potential of engineering P. pastoris into a microbial cell factory for biofuel production was evaluated in this work using (2R, 3R)-2,3-BD as an example. Engineered P. pastoris could be a promising workhorse for the production

  3. Prediction of mineral scale formation in wet gas condensate pipelines and in MEG (mono ethylene glycol) regeneration plants

    Energy Technology Data Exchange (ETDEWEB)

    Sandengen, Kristian


    Gas hydrate formation is a serious problem in the oil and gas industry, since its formation can plug wells and prevent production. The gas hydrate is a crystalline solid with a natural gas molecule surrounded by a cage of water molecules. It forms at high pressures and low temperatures. This is a problem for offshore gas wells, where the temperature is low in transport lines from well to the production facilities. Mono Ethylene Glycol (MEG) is commonly used as hydrate inhibitor. Classified as a thermodynamic inhibitor, this additive functions just as antifreeze in an automotive radiator. When producing oil and gas there will in most cases also be produced some water, which can contain dissolved salts. These salts may precipitate and they tend to deposit on surfaces. Deposition of inorganic minerals from brine is called scale. Generally MEG has the adverse effect of lowering the solubility of most salts. A common method to prevent corrosion in flow lines is to increase pH by adding basic agents (e.g. NaOH, NaHCO{sub 3}) to the MEG stream. In such cases, carbonate salts are particularly troublesome since an increase in pH by one unit, will reduce the solubility by two orders of magnitude. Thus there will be a trade off between good corrosion protection (high pH) and scale control (low pH). The aim of this work has been to develop a model that can predict mineral solubility in the presence of MEG. Experimental solubility data, together with thermodynamic data taken from literature, have been utilized to construct empirical functions for the influence of MEG on mineral scale formation. These functions enabled the expansion of an already existing aqueous scale model into a model valid for water+MEG mixed solutions. The aqueous scale model combines an equation of state (gas+oil phase) with the Pitzer ion interaction model (water phase) to describe the multiphase behaviour of gas-oil-water systems. This work summarizes the theoretical foundation and proposes how to work

  4. Producing Light Oil from a Frozen Reservoir: Reservoir and Fluid Characterization of Umiat Field, National Petroleum Reserve, Alaska

    Energy Technology Data Exchange (ETDEWEB)

    Hanks, Catherine


    with deionized water and less reduction when saturated with saline water. This reduction in relative permeability can be explained by formation of ice crystals in the center of pores. Theoretically, the radius of ice formed in the center of the pore can be determined using the Kozeny–Carman Equation by assuming the pores and pore throats as a cube with ‘N’ identical parallel pipes embedded in it. Using the values of kro obtained from the experimental work as input to the Kozeny–Carman Equation at -10ºC, the radius of ice crystals dropped from 0.145 μm to 0.069 μm when flooding-water salinity is increased to 6467 ppm. This explains the reduction of relative permeability with decreasing salinity but does not take into consideration other effects such as variations in pore throat structure. In addition, fluids like deionized water, saline water, and antifreeze (a mixture of 60% ethylene or propylene glycol with 40% water) were tested to find the best flooding agent for frozen reservoirs. At 0ºC, 9% greater recovery was observed with antifreeze was used as a flooding agent as compared to using saline water. Antifreeze showed 48% recovery even at -10ºC, at which temperature the rest of the fluids failed to increase production. Preliminary evaluation of drilling fluids indicate that the brine-based muds caused significantly less swelling in the Umiat reservoir sands when compared to fresh-water based muds. However since freezing filtrate is another cause of formation damage, a simple water-based-mud may not a viable option. It is recommended that new fluids be tested, including different salts, brines, polymers and oil-based fluids. These fluids should be tested at low temperatures in order to determine the potential for formation damage, the fluid properties under these conditions and to ensure that the freezing point is below that of the reservoir. In order to reduce the surface footprint while accessing the maximum amount of the Lower Grandstand interval

  5. Emergence of Habitable Environments in Icy World Interiors (United States)

    Neveu, Marc


    Finding habitable worlds is a key driver of solar system exploration. Many solar system missions seek environments providing liquid water, energy, and nutrients, the three ingredients necessary to sustain life [1]. Such environments include hydrothermal systems, spatially confined systems where hot aqueous fluid circulates through rock by convection. Hydrothermal activity may be widespread in the solar system. Most solar system worlds larger than 200 km in radius are icy moons and dwarf planets, likely composed of an icy, cometary mantle surrounding a rocky, chondritic core [2]. By improving an icy world evolution code [3] to include the effects of core fracturing and hydrothermal circulation, I show that several icy moons and dwarf planets likely have undergone extensive water-rock interaction [4,5]. This supports observations of aqueous products on their surfaces [6,7]. I simulated the alteration of chondritic rock [8] by pure water or fluid of cometary composition [9] to show that aqueous alteration feeds back on geophysical evolution: it modifies the fluid antifreeze content, affecting its persistence over geological timescales; and the distribution of radionuclides, whose decay is a chief heat source on dwarf planets [10]. Hydrothermal circulation also efficiently transports heat from the core into the ocean, thereby increasing ocean persistence [4]. Thus, these coupled geophysical-geochemical models provide a comprehensive picture of icy world evolution and the emergence of liquid environments in chemical disequilibrium with underlying rock in their interiors. Habitable settings also require a suitable supply of bioessential elements; but what constitutes "suitable"? I sought to quantify the bulk elemental composition of hydrothermal microbial communities, collected in hot spring sediments and mats at Yellowstone National Park, USA. To do so, one must minimize the contribution of non-biological material to the samples analyzed. This was achieved using a

  6. Traffic safety and environmental impacts. Synergies and conflicts; Liikenteen turvallisuuden ja ympaeristoevaikutusten synergiat ja vastakkainasettelut

    Energy Technology Data Exchange (ETDEWEB)

    Pollanen, M.; Ahlroth, J.; Aalto, E.; Liimatainen, H. [Tampere Univ. of Technology (Finland). Transport Research Centre Verne


    transport is an example of a synergetic measure with positive effects on both safety and the environment. Significant synergies are also achieved by promoting the renewal of vehicle stock, improving the smoothness and reliability of rail transport, reducing speed limits on highways and adopting intelligent speed adaptation systems, promoting a Single European Sky and educating, monitoring and encouraging anticipatory and economical driving habits in road and rail traffic. The greatest conflicts are related to winter traffic: the antiskid treatment of roads and streets, limiting the use of studded tyres, heating switches in rail traffic, the anti-freeze treatment of aeroplanes, and the winter maintenance of runways. Other contradictory measures include vegetation control, increasing the attractiveness of bicycling and calming traffic flow through the use of elevated pedestrian crossings and bumps. Based on the results of the study, the synergies between safety and the environment can be reinforced and the conflicts mitigated. The implementation of the most synergetic measures can be promoted by combining the perspectives of safety and the environment also in the justifications for the measures. (orig.)

  7. Understanding processes at the origin of species flocks with a focus on the marine Antarctic fauna. (United States)

    Chenuil, Anne; Saucède, Thomas; Hemery, Lenaïg G; Eléaume, Marc; Féral, Jean-Pierre; Améziane, Nadia; David, Bruno; Lecointre, Guillaume; Havermans, Charlotte


    (insularity, fragmentation and environmental stability) may favour the three criteria, although the effect is indirect for habitat dominance. We then apply this analytical framework to Antarctic marine environments by analysing data from 18 speciose clades belonging to echinoderms (five unrelated clades), notothenioid fishes (five clades) and peracarid crustaceans (eight clades). Antarctic shelf environments and history appear favourable to endemicity and speciosity, but not to ecological specialization. Two main patterns are distinguished among taxa. (i) In echinoderms, many brooding, species-rich and endemic clades are reported, but without remarkable ecological diversity or habitat dominance. In these taxa, loss of the larval stage is probably a consequence of past Antarctic environmental factors, and brooding is suggested to be responsible for enhanced allopatric speciation (via dispersal limitation). (ii) In notothenioids and peracarids, many clades fulfill all three SF criteria. This could result from unusual features in fish and crustaceans: chromosome instability and key innovations (antifreeze proteins) in notothenioids, ecological opportunity in peracarids, and a genomic architecture favouring ecological speciation in both groups. Therefore, the data do not support our starting point that normal evolutionary factors or processes drive SFs because in these two groups uncommon intrinsic features or ecological opportunity provide the best explanation. The utility of the three-criterion SF concept is therefore questioned and guidelines are given for future studies. © 2017 Cambridge Philosophical Society.

  8. Cold Fire: The Geology and Geophysics of Enceladus (United States)

    McKinnon, W. B.


    sufficient, whereas models of mantle and/or lithospheric dissipation are under active development. And what is the relation between the present activity at the SPT and the rest of the surface, most of which appears geologically youthful and also betrays evidence of high heat flows (viscously relaxed craters, groove-like extensional terrain)? Ammonia and salts have been suggested as antifreezes in Enceladus' ocean, facilitating plume eruptions. To date neither have been detected in the plumes or in the related E-ring torus, and the limits on Na are particularly stringent. The latter sets a severe constraint on the extent of water-rock interaction within Enceladus, if the proximate plume source is ocean water. An alternate view posits that the plumes are driven by degassing clathrate; this hypothesis is attractive in terms of physical chemistry but a clathrate- dominated mantle is too stiff (as presently measured) to be compatible with Enceladus' active tectonics. Whatever the cause and path for Enceladus' activity, it promises to greatly clarify our theoretical understanding of how icy worlds work.

  9. University of Iowa Cerenkov light test

    International Nuclear Information System (INIS)

    Not Available


    This Memorandum of Understanding requests beam time at Fermilab during the Winter 2005-2006 Meson Test Run to measure the signal size from various configurations of Cerenkov light generators and phototubes. The University of Iowa has pioneered in application of Cerenkov radiation for high-energy detectors. One of the results of this effort is the huge forward calorimeter for CMS (at the LHC) with a half million quartz fibers in iron. The laboratory plans to use this expertise to develop a variety of detectors making use of Cerenkov light and phototubes as the active elements in calorimeters for high-energy particles. There are three immediate applications related to forward angle calorimeters in CMS; ZDC, CASTOR, and the HE upgrade. The University of Iowa will make the detailed design for the Zero Degree Calorimeter. The proposed design uses tungsten plates interleaved with sheets of quartz fibers. In the EM part the plates are perpendicular to the beam, in the rear hadronic part they are at an angle of 45 o . Because of the limited cross sectional area of the plates, there will be considerable leakage of shower particles out of the four sides. We plan to measure this leakage and compare the results with simulations. The leakage will be measured by placing a polished aluminum tank, 4 in wide, 20 in long and 8 in deep on top of absorber material made of blocks of tantalum and copper. The fluid in the tank, that generates the Ccrcnkov light, could be water; but ethylene glycol (antifreeze, but without the added color) would be better. Being non-polar, it is not corrosive like water, and it has a higher index of refraction, 1.42 vs. 1.33. The larger index of refraction would result in more Cerenkov light and better coupling to the PMT. The University of Iowa has been asked to design the light guides for CASTOR that take the light from the quartz plates, in which the light is produced, and carry it to the PMTs. These plates, interleaved with tungsten plates, are

  10. Getting Real: A General Chemistry Laboratory Program Focusing on "Real World" Substances (United States)

    Kerber, Robert C.; Akhtar, Mohammad J.


    In order to confront the abstractness of the freshman chemistry syllabus and the consequent failure of students to relate what they learn to their everyday lives, we have designed a new freshman laboratory program. It is intended as an interface between the substances that surround the students in their ordinary lives and the abstract principles presented in chemistry classrooms (1). A laboratory should provide the organized experiences and observations that underlie the intellectual constructs of chemistry, and tying these experiences and observations to the real world can help to provide motivation for study of the principles. The freshman laboratory program constitutes the foundation for subsequent laboratory courses. However, the good habits we strive to develop there (careful observation, thorough record keeping, proper use of equipment, objective data analysis) are essential to all scientific work, and are intended to provide lasting educational value for all students, especially those who do not take later laboratory work. What We Do A list of the laboratory exercises carried out during 1994-1995 is presented in Table 1. The course incorporates the following features. 1. The exercises deal with recognizable, everyday substances, not just with "chemicals". That "baking soda" and "sodium bicarbonate" are the same is a chemical truism of which the students may be aware, but the visible presence of the Arm and Hammer box nevertheless helps them to make connections to the world outside the laboratory. Perceiving the connections, students may be inspired by curiosity to understand chemical phenomena better, not just to tolerate what they are being taught, as an irrelevant hurdle in the pursuit of a career. 2. Since many significant substances around students in the everyday world are organic, we work in the lab with organic as well as the usual inorganic materials. These include analgesics, vitamins, antifreeze, foodstuffs, dyestuffs, plastics, and fibers. In

  11. Cooption of secretory phospholipase (SPLA2) for different aspects of gravity receptor-associated mineralization in vertebrate phylogeny (United States)

    Thalmann, R.; Lu, W.


    mineral modulation not because of its enzymatic activity but to provide a rigid interface conducive to mineral interaction. To provide sufficient matrix protein for in vitro experimentation, we generated recombinant proteins. Circular dichroidism (CD) spectra indicate that the alpha helical structure of the parent SPLA2 is conserved in the SPLAL domains. A precedent of alpha helical structure for provision of a rigid interface was demonstrated to be essential for the activity of the antifreeze protein of the winter flounder. Support for alpha helical structure as signature property of the SPLAL domains of OC90 is the fact that rOC90, when exposed to calcium or carbonate-rich ionic solutions resulted in marked conformational changes, with the largest effects seen by combined application of both ions. The capacity to induce reproducible conformational changes is a testament to the quality and authenticity of rOC90. Alpha helical structure as signature characteristic of OC90 is contrary to the traditional paradigm of beta sheet structure as the essential agent in mineral interaction of highly acidic mollusk shell proteins. Apart from the alpha helical regions of the SPLAL domains, homology-based molecular modeling indicates that most of the linker segment and the terminal extensions consist of unordered structure. The significance of unordered structure in mineral interaction has recently been pointed out by several authors. For instance, the linker segment exhibits a 20 amino residue regions, dominated by hydrogen bonding and charged residues, in other words a hydrophilic segment, suitable for mineral interaction; the same applies to the C-terminal extension. Homology-based molecular models of the SPLAL domains exhibit a spherical surface with a uniform negative electrostatic potential that should be effective in attracting calcium. OC22, the principal soluble aragonitic matrix protein, consists of a single SPLAL domain, with a minor N-linked glycoside. rOC22 in vitro does

  12. The H2O2-H2O Hypothesis: Extremophiles Adapted to Conditions on Mars? (United States)

    Houtkooper, Joop M.; Schulze-Makuch, Dirk


    evolved into employing H2O2 as an antifreeze, which would also have the function as a water collector. If we would find life on Mars based on an intracellular H2O2-H2O mixture, this would not necessarily imply an independent origin of terrestrial and martian life. For that, a detailed study of the biochemistry and genetics is needed. The transfer of terrestrial organisms to Mars or vice versa is a possibility given favorable conditions for the origin and persistance of life on both planets early in solar system history (Schulze-Makuch and Houtkooper, 2007). The transfer of terrestrial organisms by early spacecrafts to Mars that either landed or crashed is a possibility, but it is not plausible that these organisms evolved in a few years. We suggest that we already have evidence of their existence from the Viking landers in two widely distant locations. The H2O2-H2O hypothesis does explain the Viking observations remarkably well, especially (1) the lack of organics detected by GC-MS, (2) the lack of detected oxidant(s) to support a chemical explanation, (3) evolution of O2 upon wetting (GEx experiment), (4) limited organic synthesis reactions (PR experiment), and (5) the gas release observations made (LR experiment)(Houtkooper and Schulze-Makuch, 2007). From the amounts of evolved CO2, O2 and N2 in the GEx experiment it can be concluded that the organisms have an excess oxidative content. This is a problem since in any destructive test, even by laser desorption-mass spectrometry (LDMS), the organisms may decompose completely into H2O, CO2, O2, and N2. The same will occur if the organisms are exposed to excess water, as they will perish due to hyperhydration. The consequence for future biology experiments is that the most fruitful approach may be the detection of metabolism under close to local environmental conditions, especially avoiding the addition of too much water. Of the Viking experiments, the PR experiment which aimed at carbon assimilation was the closest to

  13. LabVIEW-operated novel nanoliter osmometer for ice binding protein investigations. (United States)

    Braslavsky, Ido; Drori, Ran


    Ice-binding proteins (IBPs), including antifreeze proteins, ice structuring proteins, thermal hysteresis proteins, and ice recrystallization inhibition proteins, are found in cold-adapted organisms and protect them from freeze injuries by interacting with ice crystals. IBPs are found in a variety of organism, including fish(1), plants(2, 3), arthropods(4, 5), fungi(6), and bacteria(7). IBPs adsorb to the surfaces of ice crystals and prevent water molecules from joining the ice lattice at the IBP adsorption location. Ice that grows on the crystal surface between the adsorbed IBPs develops a high curvature that lowers the temperature at which the ice crystals grow, a phenomenon referred to as the Gibbs-Thomson effect. This depression creates a gap (thermal hysteresis, TH) between the melting point and the nonequilibrium freezing point, within which ice growth is arrested(8-10), see Figure 1. One of the main tools used in IBP research is the nanoliter osmometer, which facilitates measurements of the TH activities of IBP solutions. Nanoliter osmometers, such as the Clifton instrument (Clifton Technical Physics, Hartford, NY,) and Otago instrument (Otago Osmometers, Dunedin, New Zealand), were designed to measure the osmolarity of a solution by measuring the melting point depression of droplets with nanoliter volumes. These devices were used to measure the osmolarities of biological samples, such as tears(11), and were found to be useful in IBP research. Manual control over these nanoliter osmometers limited the experimental possibilities. Temperature rate changes could not be controlled reliably, the temperature range of the Clifton instrument was limited to 4,000 mOsmol (about -7.5 °C), and temperature recordings as a function of time were not an available option for these instruments. We designed a custom-made computer-controlled nanoliter osmometer system using a LabVIEW platform (National Instruments). The cold stage, described previously(9, 10), contains a metal

  14. Detection de faute automatique dans les systemes solaires thermiques basee sur la verification de regles et la simulation (United States)

    Maltais Larouche, Simon

    how the amount of work required to obtain a model could be reduced while allowing an accurate fault detection. First, the method was assessed using a calibrated model. Then, the obtained results were compared with the ones obtained with an as-built model and a model using web-based weather data instead of on-site measurements. The rule-based algorithm has successfully detected two major system faults related to a failure and replacement of the auxiliary heater and was well suited to detect instrumentation errors such as an unconnected sensor. The fault detection level has also shown to be a useful tool for fault diagnosis since it facilitates the identification of the location (which sensor), severity and time behavior of a fault. The simulation-based fault detection has allowed the detection of faults causing a performance reduction that were not always identified by the rule-based algorithm. These includes badly positioned collector's outlet temperature sensor and solar fluid freezing problem due to a too low concentration of the anti-freeze fluid. Instrumentation errors associated to some simulation inputs have sometime led to bad performance predictions from the simulation that resulted in false alarms. Fortunately, these instrument errors were always detected by the rule-based algorithm. It was also demonstrated that the performance assessment obtained with a non-calibrated model was sufficiently accurate for fault detection, since all major faults were detected without generating a significant number of false alarms. On the other hand, the model using web-based weather data was less successful and resulted in multiple false alarms. This was mainly due to an overestimation of the daily solar radiation by the weather data for days with low solar radiation. In summary, the results presented in this thesis have shown that the combination of rule-based and simulation based algorithms offers a great synergy. The conclusions obtained from this work combined with the

  15. Occurrence of benzotriazoles in the rivers Main, Hengstbach, and Hegbach (Germany). (United States)

    Kiss, Aliz; Fries, Elke


    Benzotriazoles (BT) as 1H-benzotriazole (1H-BT), 5-methyl-1H-benzotriazole (5Me-BT), and 4-methyl-1H-benzotriazole (4Me-BT) are frequently used as corrosion inhibitors in dish washer detergents, aircraft de-icing/anti-icing fluids (ADAF), automotive antifreeze formulations, brake fluids, fluids for industrial cooling systems, metal-cutting fluids, and in solid cooling lubricants. Discharge of treated municipal waste water and controlled over-runs of combined waste water sewers are potential point sources for BT in rivers. The aim of this monitoring study was to yield an overview on exposure concentrations and loads of BT in the German rivers Main, Hengstbach, and Hegbach. Concentrations of 1H-BT, 5Me-BT, and 4Me-BT were determined in grab samples collected from different sampling points in the rivers Main, Hengstbach, and Hegbach at four different sampling times. Main and Hengstbach rivers were sampled close to Frankfurt International Airport. Both rivers receive domestic waste water effluents. BT were extracted from 2.5 L of river water by solid phase extraction using Bond Elut ppl cartridges (200 mg/3 mL). The extracts were analyzed by gas chromatography/mass spectrometry in full scan mode. Mass flows of BT were calculated by concentrations multiplied by mean daily river flow rates. Median concentrations and mass flows were compared for different rivers. Mass flows were also compared for selected sampling points at different sampling times. 1H-BT, 5Me-BT, and 4Me-BT were detected in Main and Hengstbach rivers. 1H-BT and 5Me-BT were also detected in Hegbach River. Concentrations ranged from 38 to 1,474 ng/L for 1H-BT, from 25 to 281 ng/L for 5Me-BT, and from 25 to 952 ng/L for 4Me-BT. Median concentrations of 1H-BT, 5Me-BT, and 4Me-BT were lower in Main than in Hengstbach River. Much higher median mass flows of all BT were calculated for Main than for Hengstbach River. At sampling points P9 (Main) and P5 (Hengstbach) concentrations of 4Me-BT and 5Me-BT increased