Freeze-fracture study of the filamentous, segmented microorganism attached to the murine small bowel.

PubMed Central

Snellen, J E; Savage, D C

1978-01-01

A freeze-fracture study has provided new information about the filamentous, segmented microorganism known to live in the murine small bowel. The intracellular bodies produced by this microbe appear to arise by a modified sporogenesis so that they are enclosed in an envelopment membrane at least prior to release by the filament mother cell. At least some of the intracellular bodies divide while still within the mother cell, suggesting a reproductive role for these structures. The host epithelial membrane remains intact at the site of attachment, but does appear to have a reduced concentration of intramembrane particles. Changes in the host cytoplasm adjacent to the attachment site are documented and interpreted to be a sol-gel transformation which may stabilize the attachment socket. Images PMID:659364

Bioinspired Hydroxyapatite/Poly(methyl methacrylate) Composite with a Nacre-Mimetic Architecture by a Bidirectional Freezing Method.

PubMed

Bai, Hao; Walsh, Flynn; Gludovatz, Bernd; Delattre, Benjamin; Huang, Caili; Chen, Yuan; Tomsia, Antoni P; Ritchie, Robert O

2016-01-06

Using a bidirectional freezing technique, combined with uniaxial pressing and in situ polymerization, "nacre-mimetic" hydroxyapatite/poly(methyl methacrylate) (PMMA) composites are developed by processing large-scale aligned lamellar ceramic scaffolds. Structural and mechanical characterization shows "brick-and-mortar" structures, akin to nacre, with interesting combinations of strength, stiffness, and work of fracture, which provide a pathway to making strong and tough lightweight materials. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Assessment of different sample preparation routes for mass spectrometric monitoring and imaging of lipids in bone cells via ToF-SIMS

PubMed Central

Schaepe, Kaija; Kokesch-Himmelreich, Julia; Rohnke, Marcus; Wagner, Alena-Svenja; Schaaf, Thimo; Wenisch, Sabine; Janek, Jürgen

2015-01-01

In ToF-SIMS analysis, the experimental outcome from cell experiments is to a great extent influenced by the sample preparation routine. In order to better judge this critical influence in the case of lipid analysis, a detailed comparison of different sample preparation routines is performed—aiming at an optimized preparation routine for systematic lipid imaging of cell cultures. For this purpose, human mesenchymal stem cells were analyzed: (a) as chemically fixed, (b) freeze-dried, and (c) frozen-hydrated. For chemical fixation, different fixatives, i.e., glutaraldehyde, paraformaldehyde, and a mixture of both, were tested with different postfixative handling procedures like storage in phosphate buffered saline, water or critical point drying. Furthermore, secondary lipid fixation via osmium tetroxide was taken into account and the effect of an ascending alcohol series with and without this secondary lipid fixation was evaluated. Concerning freeze-drying, three different postprocessing possibilities were examined. One can be considered as a pure cryofixation technique while the other two routes were based on chemical fixation. Cryofixation methods known from literature, i.e., freeze-fracturing and simple frozen-hydrated preparation, were also evaluated to complete the comparison of sample preparation techniques. Subsequent data evaluation of SIMS spectra in both, positive and negative, ion mode was performed via principal component analysis by use of peak sets representative for lipids. For freeze-fracturing, these experiments revealed poor reproducibility making this preparation route unsuitable for systematic investigations and statistic data evaluation. Freeze-drying after cryofixation showed improved reproducibility and well preserved lipid contents while the other freeze-drying procedures showed drawbacks in one of these criteria. In comparison, chemical fixation techniques via glutar- and/or paraformaldehyde proved most suitable in terms of reproducibility and preserved lipid contents, while alcohol and osmium treatment led to the extraction of lipids and are therefore not recommended. PMID:25791294

Assessment of different sample preparation routes for mass spectrometric monitoring and imaging of lipids in bone cells via ToF-SIMS.

PubMed

Schaepe, Kaija; Kokesch-Himmelreich, Julia; Rohnke, Marcus; Wagner, Alena-Svenja; Schaaf, Thimo; Wenisch, Sabine; Janek, Jürgen

2015-03-19

In ToF-SIMS analysis, the experimental outcome from cell experiments is to a great extent influenced by the sample preparation routine. In order to better judge this critical influence in the case of lipid analysis, a detailed comparison of different sample preparation routines is performed-aiming at an optimized preparation routine for systematic lipid imaging of cell cultures. For this purpose, human mesenchymal stem cells were analyzed: (a) as chemically fixed, (b) freeze-dried, and (c) frozen-hydrated. For chemical fixation, different fixatives, i.e., glutaraldehyde, paraformaldehyde, and a mixture of both, were tested with different postfixative handling procedures like storage in phosphate buffered saline, water or critical point drying. Furthermore, secondary lipid fixation via osmium tetroxide was taken into account and the effect of an ascending alcohol series with and without this secondary lipid fixation was evaluated. Concerning freeze-drying, three different postprocessing possibilities were examined. One can be considered as a pure cryofixation technique while the other two routes were based on chemical fixation. Cryofixation methods known from literature, i.e., freeze-fracturing and simple frozen-hydrated preparation, were also evaluated to complete the comparison of sample preparation techniques. Subsequent data evaluation of SIMS spectra in both, positive and negative, ion mode was performed via principal component analysis by use of peak sets representative for lipids. For freeze-fracturing, these experiments revealed poor reproducibility making this preparation route unsuitable for systematic investigations and statistic data evaluation. Freeze-drying after cryofixation showed improved reproducibility and well preserved lipid contents while the other freeze-drying procedures showed drawbacks in one of these criteria. In comparison, chemical fixation techniques via glutar- and/or paraformaldehyde proved most suitable in terms of reproducibility and preserved lipid contents, while alcohol and osmium treatment led to the extraction of lipids and are therefore not recommended.

Mechanical Behavior of Tough Hydrogels for Structural Applications

NASA Astrophysics Data System (ADS)

Illeperuma, Widusha Ruwangi Kaushalya

Hydrogels are widely used in many commercial products including Jell-O, contact lenses, and superabsorbent diapers. In recent decades, hydrogels have been under intense development for biomedical applications, such as scaffolds in tissue engineering, carriers for drug delivery, and valves in microfluidic systems. But the scope is severely limited as conventional hydrogels are weak and brittle and are not very stretchable. This thesis investigates the approaches that enhance the mechanical properties of hydrogels and their structural applications. We discov¬ered a class of exceptionally stretchable and tough hydrogels made from poly-mers that form networks via ionic and covalent crosslinks. Although such a hydrogel contains ~90% water, it can be stretched beyond 20 times its initial length, and has a fracture energy of ~9000 J/m2. The combination of large stretchability, remarkable toughness, and recoverability of stiffness and toughness, along with easy synthesis makes this material much superior over existing hydrogels. Extreme stretchability and blunted crack tips of these hydrogels question the validity of traditional fracture testing methods. We re-examine a widely used pure shear test method to measure the fracture energy. With the experimental and simulation results, we conclude that the pure shear test method can be used to measure fracture energy of extremely stretchable materials. Even though polyacrylamide-alginate hydrogels have an extremely high toughness, it has a relatively low stiffness and strength. We improved the stiffness and strength by embedding fibers. Most hydrogels are brittle, allowing the fibers to cut through the hydrogel when the composite is loaded. But tough hydrogel composites do not fail by the fibers cutting the hydrogel; instead, it undergoes large deforming by fibers sliding through the matrix. Hydrogels were not considered as materials for structural applications. But with enhanced mechanical properties, they have opened up novel applications. This thesis aims to investigate the broader applications, well beyond those investigated so far. We show fiber reinforced tough hydrogels can dissipate a significant amount of energy at a tunable level of stress, making them suitable for energy absorbing applications such as inner layer of helmets. We develop inexpensive fire-retarding materials using tough hydrogels that provide superior protection from burn injuries. We also study hydrogels as actuators that can be used in soft robotics. Hydrogels contain mostly water and they freeze when the temperature drops below 00C and lose its functions. We demonstrate a new class of hydrogels that do not freeze and hydrogels that partially freeze below water freezing temperature. Partially freezing hydrogels are ideal for cooling applications such as gel packs and non-freezing hydrogels are useful in all the structural applications at low temperatures. This thesis will enable the use of inexpensive hydrogels in a new class of non-traditional structural applications where the mechanical behavior of the hydrogel is of prime importance.

The Effect of Freezing on the Dynamics of Dike Propagation

NASA Astrophysics Data System (ADS)

Tait, S.; Taisne, B.

2007-12-01

When magma-filled cracks propagate close to the Earth's surface, host rock temperature is well below the magma solidus. Solidification and substantial increase in magma viscosity can occur, are most pronounced near the propagating tip and can slow or arrest the progress of the dike. Quantitative analysis is required to predict whether a given dike will reach the surface to erupt and the duration of the precursor sequence. This challenging physical problem mixes elasticity, fracture mechanics, heat transfer and fluid flow with strong rheologic gradients due to cooling. We describe the propagation behaviour of such a hydraulic fracture using a laboratory experimental system of a crack fed by a constant flux of paraffin wax from a source reservoir propagating through gelatin below the solidus of the wax. The most novel behaviour is an intermittent regime in which cracks periodically stop advancing due to solidification, then swell at constant length while enhancing the elastic deformation in the surrounding solid before propagation resumes. We present a physical model of this system, based on different balances between driving and resistive forces: the former are elastic stress and liquid buoyancy, the latter are fracture resistance at the tip and viscous resistance. The fracture is represented as a head, behind the propagating tip, connected to the source by a narrow tail. Freezing of liquid close to the tip is assumed to enhance fracture resistance according to a cooling law, and propagation is assumed to occur only when the stress exerted by the liquid is enough to overcome fracture resistance. Our theoretical model reproduces intermittent propagation with precise behaviour depending on the controlling stress balances, and provides a tool to analyse natural systems.

Freeze-fracture studies of photoreceptor membranes: new observations bearing upon the distribution of cholesterol

PubMed Central

1983-01-01

We performed electron microscopy of replicas from freeze-fractured retinas exposed during or after fixation to the cholesterol-binding antibiotic, filipin. We observed characteristic filipin-induced perturbations throughout the disk and plasma membranes of retinal rod outer segments of various species. It is evident that a prolonged exposure to filipin in fixative enhances rather than reduces presumptive cholesterol detection in the vertebrate photoreceptor cell. In agreement with the pattern seen in our previous study (Andrews, L.D., and A. I. Cohen, 1979, J. Cell Biol., 81:215-228), filipin- binding in membranes exhibiting particle-free patches seemed largely confined to these patches. Favorably fractured photoreceptors exhibited marked filipin-binding in apical inner segment plasma membrane topologically confluent with and proximate to the outer segment plasma membrane, which was comparatively free of filipin binding. A possible boundary between these differing membrane domains was suggested in a number of replicas exhibiting lower filipin binding to the apical plasma membrane of the inner segment in the area surrounding the cilium. This area contains a structure (Andrews, L. D., 1982, Freeze- fracture studies of vertebrate photoreceptors, In Structure of the Eye, J. G. Hollyfield and E. Acosta Vidrio, editors, Elsevier/North-Holland, New York, 11-23) that resembles the active zones of the nerve terminals for the frog neuromuscular junction. These observations lead us to hypothesize that these structures may function to direct vesicle fusion to occur near them, in a domain of membrane more closely resembling outer than inner segment plasma membrane. The above evidence supports the views that (a) all disk membranes contain cholesterol, but the particle-free patches present in some disks trap cholesterol from contiguous particulate membrane regions; (b) contiguous inner and outer segment membranes may greatly differ in cholesterol content; and (c) the suggested higher cholesterol in the inner segment than in the outer segment plasma membrane may help direct newly inserted photopigment molecules to the outer segment. PMID:6411740

Scanning electron microscopy of high-pressure-frozen sea urchin embryos.

PubMed

Walther, P; Chen, Y; Malecki, M; Zoran, S L; Schatten, G P; Pawley, J B

1993-12-01

High-pressure-freezing permits direct cryo-fixation of sea urchin embryos having a defined developmental state without the formation of large ice crystals. We have investigated preparation protocols for observing high-pressure-frozen and freeze-fractured samples in the scanning electron microscope. High-pressure-freezing was superior to other freezing protocols, because the whole bulk sample was reasonably well frozen and the overall three-dimensional shape of the embryos was well preserved. The samples were either dehydrated by freeze-substitution and critical-point-drying, or imaged in the partially hydrated state, using a cold stage in the SEM. During freeze-substitution the samples were stabilized by fixatives. The disadvantage of this method was that shrinking and extraction effects, caused by the removal of the water, could not be avoided. These disadvantages were avoided when the sample was imaged in the frozen-hydrated state using a cold-stage in the SEM. This would be the method of choice for morphometric studies. Frozen-hydrated samples, however, were very beam sensitive and many structures remained covered by the ice and were not visible. Frozen-hydrated samples were partially freeze-dried to make visible additional structures that had been covered by ice. However, this method also caused drying artifacts when too much water was removed.

A novel adhering junction in the apical ciliary apparatus of the rotifer Brachionus plicatilis (Rotifera, Monogononta).

PubMed

Dallai, R; Lupetti, P; Lane, N J

1996-10-01

Cultures of the rotifer Brachionus plicatilis were examined with regard to their interepithelial junctions after infiltration with the extracellular tracer lanthanum, freeze-fracturing or quick-freeze deep-etching. The lateral borders between ciliated cells have an unusual apical adhering junction. This apical part of their intercellular cleft looks desmosome-like, but it is characterized by unusual intramembranous E-face clusters of particles. Deep-etching reveals that these are packed together in short rows which lie parallel to one another in orderly arrays. The true membrane surface in these areas features filaments in the form of short ribbons; these are produced by projections, possibly part of the glycocalyx, emerging from the membranes, between which the electron-dense tracer lanthanum permeates. These projections appear to overlap with each other in the centre of the intercellular cleft; this would provide a particularly flexible adaptation to maintain cell-cell contact and coordination as a consequence. The filamentous ribbons may be held in position by the intramembranous particle arrays since both have a similar size and distribution. These contacts are quite different from desmosomes and appear to represent a distinct new category of adhesive cell-cell junction. Beneath these novel structures, conventional pleated septate junctions are found, exhibiting the undulating intercellular ribbons typical of this junctional type, as well as the usual parallel alignments of intramembranous rows of EF grooves and PF particles. Below these are found gap junctions as close-packed plaques of intramembranous particles on either the P-face or E-face. After freeze-fracturing, the complementary fracture face to the particles shows pits, usually on the P-face, arrayed with a very precise hexagonal pattern.

Structure of biodiesel based bicontinuous microemulsions for environmentally compatible decontamination: A small angle neutron scattering and freeze fracture electron microscopy study.

PubMed

Wellert, S; Karg, M; Imhof, H; Steppin, A; Altmann, H-J; Dolle, M; Richardt, A; Tiersch, B; Koetz, J; Lapp, A; Hellweg, T

2008-09-01

Most toxic industrial chemicals and chemical warfare agents are hydrophobic and can only be solubilized in organic solvents. However, most reagents employed for the degradation of these toxic compounds can only be dissolved in water. Hence, microemulsions are auspicious media for the decontamination of a variety of chemical warfare agents and pesticides. They allow for the solubilization of both the lipophilic toxics and the hydrophilic reagent. Alkyl oligoglucosides and plant derived solvents like rapeseed methyl ester enable the formulation of environmentally compatible bicontinuous microemulsions. In the present article the phase behavior of such a microemulsion is studied and the bicontinuous phase is identified. Small angle neutron scattering (SANS) and freeze fracture electron microscopy (FFEM) measurements are used to characterize the structure of the bicontinuous phase and allow for an estimation of the total internal interface. Moreover, also the influence of the co-surfactant (1-pentanol) on the structural parameters of the bicontinuous phase is studied with SANS.

The plasma membrane of myxosporidian valve cells: freeze fracture data.

PubMed

Desportes-Livage, I; Nicolas, G

1990-01-01

Freeze fracturing of Myxosporidian spores reveals the occurrence of a continuous layer of transmembrane particles all over the surface area of the valve cells which form the spore envelope. These particles are densely packed all over the P face membrane. Due to their polygonal outline, their diameter (6-7 nm) and their central core, they resemble the particles forming the connections of gap junctions which metabolically couple the neighboring cells in animal tissues. In the present report, the role of the transmembrane particles is still hypothetical. However, they might represent a membrane structural specialization of the spores which are submitted to osmotic variations of the fluid external medium. Furthermore similar transmembrane particles are observed at the level of the septate junction which seals the valve cells. In this occurrence, they are arranged in a series of 40 double rows parallel to the suture of the spore envelope. These findings support the view that Myxosporidia are Metazoa and raise the problem of their origin.

Dissolved Gases and Ice Fracturing During the Freezing of a Multicellular Organism: Lessons from Tardigrades

PubMed Central

Kletetschka, Gunther; Hruba, Jolana

2015-01-01

Abstract Three issues are critical for successful cryopreservation of multicellular material: gases dissolved in liquid, thermal conductivity of the tissue, and localization of microstructures. Here we show that heat distribution is controlled by the gas amount dissolved in liquids and that when changing the liquid into solid, the dissolved gases either form bubbles due to the absence of space in the lattice of solids and/or are migrated toward the concentrated salt and sugar solution at the cost of amount of heat required to be removed to complete a solid-state transition. These factors affect the heat distribution in the organs to be cryopreserved. We show that the gas concentration issue controls fracturing of ice when freezing. There are volumetric changes not only when changing the liquid into solid (volume increases) but also reduction of the volume when reaching lower temperatures (volume decreases). We discuss these issues parallel with observations of the cryosurvivability of multicellular organisms, tardigrades, and discuss their analogy for cryopreservation of large organs. PMID:26309797

Dissolved Gases and Ice Fracturing During the Freezing of a Multicellular Organism: Lessons from Tardigrades.

PubMed

Kletetschka, Gunther; Hruba, Jolana

2015-01-01

Three issues are critical for successful cryopreservation of multicellular material: gases dissolved in liquid, thermal conductivity of the tissue, and localization of microstructures. Here we show that heat distribution is controlled by the gas amount dissolved in liquids and that when changing the liquid into solid, the dissolved gases either form bubbles due to the absence of space in the lattice of solids and/or are migrated toward the concentrated salt and sugar solution at the cost of amount of heat required to be removed to complete a solid-state transition. These factors affect the heat distribution in the organs to be cryopreserved. We show that the gas concentration issue controls fracturing of ice when freezing. There are volumetric changes not only when changing the liquid into solid (volume increases) but also reduction of the volume when reaching lower temperatures (volume decreases). We discuss these issues parallel with observations of the cryosurvivability of multicellular organisms, tardigrades, and discuss their analogy for cryopreservation of large organs.

Colloid-facilitated mobilization of metals by freeze-thaw cycles.

PubMed

Mohanty, Sanjay K; Saiers, James E; Ryan, Joseph N

2014-01-21

The potential of freeze-thaw cycles to release colloids and colloid-associated contaminants into water is unknown. We examined the effect of freeze-thaw cycles on the mobilization of cesium and strontium in association with colloids in intact cores of a fractured soil, where preferential flow paths are prevalent. Two intact cores were contaminated with cesium and strontium. To mobilize colloids and metal cations sequestered in the soil cores, each core was subjected to 10 intermittent wetting events separated by 66 h pauses. During the first five pauses, the cores were dried at room temperature, and during last five pauses, the cores were subjected to 42 h of freezing followed by 24 h of thawing. In comparison to drying, freeze-thaw cycles created additional preferential flow paths through which colloids, cesium, and strontium were mobilized. The wetting events following freeze-thaw intervals mobilized about twice as many colloids as wetting events following drying at room temperature. Successive wetting events following 66 h of drying mobilized similar amounts of colloids; in contrast, successive wetting events after 66 h of freeze-thaw intervals mobilized greater amounts of colloids than the previous one. Drying and freeze-thaw treatments, respectively, increased and decreased the dissolved cesium and strontium, but both treatments increased the colloidal cesium and strontium. Overall, the freeze-thaw cycles increased the mobilization of metal contaminants primarily in association with colloids through preferential flow paths. These findings suggest that the mobilization of colloid and colloid-associated contaminants could increase when temperature variations occur around the freezing point of water. Thus, climate extremes have the potential to mobilize contaminants that have been sequestered in the vadose zone for decades.

Freeze-Fracture Ultrastructure of Thylakoid Membranes in Chloroplasts from Manganese-Deficient Plants

PubMed Central

Simpson, David J.; Robinson, Simon P.

1984-01-01

Leaves from spinach (Spinacia oleracea L. cv Hybrid 102) plants grown in Mn-deficient nutrient solution were characterized by chlorosis, lowered chlorophyll a/b ratio and reduced electron transport. There were characteristic changes in room temperature fluorescence induction kinetics with increased initial yield (Fo) and decreased variable fluorescence (Fv). The fluorescence yield after the maximum fell rapidly to a level below Fo. The shape of the rise from Fo to the maximum was altered and the size of photosystem II units increased, as measured by half-rise time of Fv in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. The Mn-deficient leaves were harvested before necrosis, when thin section electron microscopy revealed no disorganization of the thylakoid system. Thylakoid membranes were examined by freeze-fracture electron microscopy. The effect of Mn-deficiency was the specific loss of three-quarters of the particles from the endoplasmic fracture face of appressed thylakoids (EFs). Mn-deficient leaves were restored to near normal 2 days after application of exogenous Mn to the nutrient solution. It is concluded that the loss of most, but not all, functional photosystem II reaction centers from grana, with no alteration in light-harvesting complex or photosystem I, is responsible for the fluorescence and functional properties observed. The response of thylakoids to Mn deficiency shows that there is a fundamental difference in composition and function of stacked and unstacked endoplasmic fracture particles. The stacked endoplasmic fracture particle probably contains, in close association, the photosystem II reaction center and also the Mn-containing polypeptide, the 3-(3,4-dichlorophenyl)-1,1-dimethylurea-binding protein, and all electron transport components in between. Images Fig. 3 Fig. 4 Fig. 5 PMID:16663491

A Periglacial Analog for Landforms in Gale Crater, Mars

NASA Technical Reports Server (NTRS)

Oehler, Dorothy Z.

2013-01-01

Several features in a high thermal inertia (TI) unit at Gale crater can be interpreted within a periglacial framework. These features include polygonally fractured terrain (cf. ice-wedge polygons), circumferential patterns of polygonal fractures (cf. relict pingos with ice-wedge polygons on their surfaces), irregularly-shaped and clustered depressions (cf. remnants of collapsed pingos and ephemeral lakes), and a general hummocky topography (cf. thermokarst). This interpretation would imply a major history of water and ice in Gale crater, involving permafrost, freeze-thaw cycles, and perhaps ponded surface water.

Freeze fracturing of elastic porous media: a mathematical model

PubMed Central

Vlahou, I.; Worster, M. G.

2015-01-01

We present a mathematical model of the fracturing of water-saturated rocks and other porous materials in cold climates. Ice growing inside porous rocks causes large pressures to develop that can significantly damage the rock. We study the growth of ice inside a penny-shaped cavity in a water-saturated porous rock and the consequent fracturing of the medium. Premelting of the ice against the rock, which results in thin films of unfrozen water forming between the ice and the rock, is one of the dominant processes of rock fracturing. We find that the fracture toughness of the rock, the size of pre-existing faults and the undercooling of the environment are the main parameters determining the susceptibility of a medium to fracturing. We also explore the dependence of the growth rates on the permeability and elasticity of the medium. Thin and fast-fracturing cracks are found for many types of rocks. We consider how the growth rate can be limited by the existence of pore ice, which decreases the permeability of a medium, and propose an expression for the effective ‘frozen’ permeability. PMID:25792954

Freeze fracturing of elastic porous media: a mathematical model.

PubMed

Vlahou, I; Worster, M G

2015-03-08

We present a mathematical model of the fracturing of water-saturated rocks and other porous materials in cold climates. Ice growing inside porous rocks causes large pressures to develop that can significantly damage the rock. We study the growth of ice inside a penny-shaped cavity in a water-saturated porous rock and the consequent fracturing of the medium. Premelting of the ice against the rock, which results in thin films of unfrozen water forming between the ice and the rock, is one of the dominant processes of rock fracturing. We find that the fracture toughness of the rock, the size of pre-existing faults and the undercooling of the environment are the main parameters determining the susceptibility of a medium to fracturing. We also explore the dependence of the growth rates on the permeability and elasticity of the medium. Thin and fast-fracturing cracks are found for many types of rocks. We consider how the growth rate can be limited by the existence of pore ice, which decreases the permeability of a medium, and propose an expression for the effective 'frozen' permeability.

Comparison of drying characteristic and uniformity of banana cubes dried by pulse-spouted microwave vacuum drying, freeze drying and microwave freeze drying.

PubMed

Jiang, Hao; Zhang, Min; Mujumdar, Arun S; Lim, Rui-Xin

2014-07-01

To overcome the flaws of high energy consumption of freeze drying (FD) and the non-uniform drying of microwave freeze drying (MFD), pulse-spouted microwave vacuum drying (PSMVD) was developed. The results showed that the drying time can be dramatically shortened if microwave was used as the heating source. In this experiment, both MFD and PSMVD could shorten drying time by 50% as compared to the FD process. Depending on the heating method, MFD and PSMVD dried banana cubes showed trends of expansion while FD dried samples demonstrated trends of shrinkage. Shrinkage also brought intensive structure and highest fracturability of all three samples dried by different methods. The residual ascorbic acid content of PSMVD dried samples can be as high as in FD dried samples, which were superior to MFD dried samples. The tests confirmed that PSMVD could bring about better drying uniformity than MFD. Besides, compared with traditional MFD, PSMVD can provide better extrinsic feature, and can bring about improved nutritional features because of the higher residual ascorbic acid content. © 2013 Society of Chemical Industry.

Fabrication and characterisation of a novel biomimetic anisotropic ceramic/polymer-infiltrated composite material.

PubMed

Al-Jawoosh, Sara; Ireland, Anthony; Su, Bo

2018-04-10

To fabricate and characterise a novel biomimetic composite material consisting of aligned porous ceramic preforms infiltrated with polymer. Freeze-casting was used to fabricate and control the microstructure and porosity of ceramic preforms, which were subsequently infiltrated with 40-50% by volume UDMA-TEGDMA polymer. The composite materials were then subjected to characterisation, namely density, compression, three-point bend, hardness and fracture toughness testing. Samples were also subjected to scanning electron microscopy and computerised tomography (Micro-CT). Three-dimensional aligned honeycomb-like ceramic structures were produced and full interpenetration of the polymer phase was observed using micro-CT. Depending on the volume fraction of the ceramic preform, the density of the final composite ranged from 2.92 to 3.36g/cm 3 , compressive strength ranged from 206.26 to 253.97MPa, flexural strength from 97.73 to 145.65MPa, hardness ranged from 1.46 to 1.62GPa, and fracture toughness from 3.91 to 4.86MPam 1/2 . Freeze-casting provides a novel method to engineer composite materials with a unique aligned honeycomb-like interpenetrating structure, consisting of two continuous phases, inorganic and organic. There was a correlation between the ceramic fraction and the subsequent, density, strength, hardness and fracture toughness of the composite material. Copyright © 2018 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.

Failure of cement hydrates: freeze-thaw and fracture

NASA Astrophysics Data System (ADS)

Ioannidou, Katerina; Del Gado, Emanuela; Ulm, Franz-Josef; Pellenq, Roland

Mechanical and viscoelastic behavior of concrete crucially depends on cement hydrates, the ``glue'' of cement. Even more than the atomistic structure, the mesoscale amorphous texture of cement hydrates over hundreds of nanometers plays a crucial role for material properties. We use simulations that combine information of the nano-scale building units of cement hydrates and on their effective interactions, obtained from atomistic simulations and experiments, into a statistical physics framework for aggregating nanoparticles.Our mesoscale model was able to reconcile different experimental results ranging from small-angle neutron scattering, SEM, adsorption/desorption of N2, and water to nanoindentation and gain the new fundamental insights into the microscopic origin of the properties measured. Our results suggest that heterogeneities developed during the early stages of hydration persist in the structure of C-S-H, impacting the rheological and mechanical performance of the hardened cement paste. In this talk I discuss recent investigation on failure mechanism at the mesoscale of hardened cement paste such as freeze-thaw and fracture. Using correlations between local volume fractions and local stress we provide a link between structural and mechanical heterogeneities during the failure mechanisms.

Structure of the cell envelope of corynebacteria: importance of the non-covalently bound lipids in the formation of the cell wall permeability barrier and fracture plane.

PubMed

Puech, V; Chami, M; Lemassu, A; Lanéelle, M A; Schiffler, B; Gounon, P; Bayan, N; Benz, R; Daffé, M

2001-05-01

With the recent success of the heterologous expression of mycobacterial antigens in corynebacteria, in addition to the importance of these bacteria in biotechnology and medicine, a better understanding of the structure of their cell envelopes was needed. A combination of molecular compositional analysis, ultrastructural appearance and freeze-etch electron microscopy study was used to arrive at a chemical model, unique to corynebacteria but consistent with their phylogenetic relatedness to mycobacteria and other members of the distinctive suprageneric actinomycete taxon. Transmission electron microscopy and chemical analyses showed that the cell envelopes of the representative strains of corynebacteria examined consisted of (i) an outer layer composed of polysaccharides (primarily a high-molecular-mass glucan and arabinomannans), proteins, which include the mycoloyltransferase PS1, and lipids; (ii) a cell wall glycan core of peptidoglycan-arabinogalactan which may contain other sugar residues and was usually esterified by corynomycolic acids; and (iii) a typical plasma membrane bilayer. Freeze-etch electron microscopy showed that most corynomycolate-containing strains exhibited a main fracture plane in their cell wall and contained low-molecular-mass porins, while the fracture occurred within the plasma membrane of strains devoid of both corynomycolate and pore-forming proteins. Importantly, in most strains, the amount of cell wall-linked corynomycolates was not sufficient to cover the bacterial surface; interestingly, the occurrence of a cell wall fracture plane correlated with the amount of non-covalently bound lipids of the strains. Furthermore, these lipids were shown to spontaneously form liposomes, indicating that they may participate in a bilayer structure. Altogether, the data suggested that the cell wall permeability barrier in corynebacteria involved both covalently linked corynomycolates and non-covalently bound lipids of their cell envelopes.

The USAF Phillips Laboratory sodium-sulfur battery technology program: Results and status

NASA Technical Reports Server (NTRS)

Rainbow, Marc E.; Somerville, Andrew

1996-01-01

Tests performed on NaS batteries are reported. The results of safety and abuse testing, shock and vibration tests, cell failure on warm-up, freeze thaw, overtemperature conditions, electrolyte fracture, overdischarge, and short circuit tests are presented along with GEO and LEO cycle tests and the status of the NaS cell flight tests.

Adhesion between liposomes mediated by the chlorophyll a/b light- harvesting complex isolated from chloroplast membranes

PubMed Central

1980-01-01

A highly purified chlorophyll a/b light-harvesting complex (chl a/b LHC; chl a/b ratio 1.2) was obtained from Triton-solubilized chloroplast membranes of pea and barley according to the method of Burke et al. (1978, Arch. Biochem. Biophys. 187: 252--263). Gel electrophoresis of the cation-precipitated chl a/b LHC from peas reveals the presence of four polypeptides in the 23- to 28-kdalton size range. Three of these peptides appear to be identical to those derived from re-electrophoresed CPII and CPII* bands. In freeze-fracture replicas, the cation-precipitated chl a/b LHC appears as a semicrystalline aggregate of membranous sheets containing closely spaced granules. Upon removal of the cations by dialysis, the aggregates break up into their constituent membranous sheets without changing their granular substructure. These membranous sheets can be resolubilized in 1.5% Triton X-100, and the chl a/b LHC particles then reconstituted into soybean lecithin liposomes. Freeze-fracture micrographs of the reconstituted chl a/b LHC vesicles suspended in a low salt medium reveal randomly dispersed approximately 80-A particles on both concave and convex fracture faces as well as some crystalline particle arrays, presumably resulting from incompletely solubilized fragments of the membranous sheets. Based on the approximately 80-A diameter of the particles, and on the assumption that one freeze- fracture particle represents the structural unit of one chl a/b LHC aggregate, a theoretical mol wt of approximately 200 kdalton has been calculated for the chl a/b LHC. Deep-etching and negative-staining techniques reveal that the chl a/b LHC particles are also exposed on the surface of the bilayer membranes. Addition of greater than or equal to 2 mM MgCl2 or greater than or equal to 60 mM NaCl to the reconstituted vesicles leads to their aggregation and, with divalent cations, to the formation of extensive membrane stacks. At the same time, the chl a/b LHC particles become clustered into the adhering membrane regions. Under these conditions the particles in adjacent membranes usually become precisely aligned. Evidence is presented to aupport the hypothesis that adhesion between the chl a/b LHC particles is mediated by hydrophobic interactions, and that the cations are needed to neutralize surface charges on the particles. PMID:7350170

Rockfall triggering by cyclic thermal stressing of exfoliation fractures

USGS Publications Warehouse

Collins, Brian D.; Stock, Greg M.

2016-01-01

Exfoliation of rock deteriorates cliffs through the formation and subsequent opening of fractures, which in turn can lead to potentially hazardous rockfalls. Although a number of mechanisms are known to trigger rockfalls, many rockfalls occur during periods when likely triggers such as precipitation, seismic activity and freezing conditions are absent. It has been suggested that these enigmatic rockfalls may occur due to solar heating of rock surfaces, which can cause outward expansion. Here we use data from 3.5 years of field monitoring of an exfoliating granite cliff in Yosemite National Park in California, USA, to assess the magnitude and temporal pattern of thermally induced rock deformation. From a thermodynamic analysis, we find that daily, seasonal and annual temperature variations are sufficient to drive cyclic and cumulative opening of fractures. Application of fracture theory suggests that these changes can lead to further fracture propagation and the consequent detachment of rock. Our data indicate that the warmest times of the day and year are particularly conducive to triggering rockfalls, and that cyclic thermal forcing may enhance the efficacy of other, more typical rockfall triggers.

Environmental controls on micro fracture processes in shelf ice

NASA Astrophysics Data System (ADS)

Sammonds, Peter

2013-04-01

The recent retreat and collapse of the ice shelves on the Antarctic Peninsula has been associated with regional atmospheric warming, oceanic warming, increased summer melt and shelf flexure. Although the cause of collapse is a matter of active discussion, the process is that of fracture of a creep-brittle material, close to its melting point. The environmental controls on how fracturing initiates, at a micro-scale, strongly determine the macroscopic disintegration of ice shelves. In particular the shelf temperature profile controls the plasticity of the ice shelf; the densification of shelf ice due to melting and re-freezing affects the crack tip stress intensity; the accretion of marine ice at the bottom of the shelf imposes a thermal/mechanical discontinuity; saline environments control crack tip stress corrosion; cyclic loading promotes sub-critical crack propagation. These strong environmental controls on shelf ice fracture means that assessing shelf stability is a non-deterministic problem. How these factors may be parameterized in ice shelf models, through the use of fracture mechanisms maps, is discussed. The findings are discussed in relation to the stability of Larsen C.

Molecular morphology of the tetrodotoxin-binding sodium channel protein from Electrophorus electricus in solubilized and reconstituted preparations

PubMed Central

1983-01-01

The appearance of detergent-solubilized voltage-regulated sodium channel protein was recently characterized by this laboratory. Negative- staining revealed rod-shaped particles measuring 40 X 170 A. Further studies have suggested that the actual configuration of this protein may be quite different from the rod-shaped structures. Freeze-fracture and freeze-etch images of the protein in reconstituted membranes indicated that the channel is cylindrical with a diameter of 100 A and a minimum length of 80 A. Experiments with two detergent systems (Lubrol-PX and sodium cholate) enabled us to explain the discrepancy between this structure and the rod-shaped particles visualized earlier. Negative staining in either detergent at low pH (4.5) produced rod- shaped structures. As the pH was increased, doughnut-shaped particles, consistent with the structure of the protein in freeze-etch, appeared in negative stain. The tendency of the protein to change shape under different pH conditions appears to be a peculiar property of this protein. PMID:6315745

Plant cell plasma membrane structure and properties under clinostatting

NASA Astrophysics Data System (ADS)

Polulakh, Yu. A.; Zhadko, S. I.; Klimchuk, D. A.; Baraboy, V. A.; Alpatov, A. N.; Sytnik, K. M.

Structural-functional organization of plasma membrane of pea roots seedling was investigated by methods of chemiluminescence, fluorescence probes, chromatography and freeze-fracture studies under normal conditions and clinostatting. Phase character of lipid peroxidation intensity was fixed. The initial phase of this process is characterized by lipid peroxidation decreasing with its next induction. The primary changes depending on free-radical mechanisms of lipid peroxidation were excellently revealed by chemiluminescence. Plasmalemma microviscosity increased on the average of 15-20 % under microgravity at the initial stages of its phenomenon. There were major changes of phosphatidilcholine and phosphatidilethanolamine contents. The total quantity of phospholipids remained rather stable. Changes of phosphatide acid concentration point to degradation and phospholipids biosynthesis. There were increases of unsaturated fatty acids mainly at the expense of linoleic and linolenic acids and also a decrease of saturated fatty acid content at the expense of palmitic and stearic acids. Unsaturation index of fatty acids increased as well. On the whole fatty acid composition was variable in comparison with phospholipids. Probably it is one of mechanisms of maintaining of microviscosity within definite limits. Considerable structural changes in organization of plasmalemma protein-lipid complex were not revealed by the freeze-fracture studies.

Freeze-fracture scanning electron microscopy of Lemna minor L. (duckweed)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Echlin, P.; Pawley, J.B.; Hayes, T.L.

1979-01-01

A detailed study has been carried out on the frozen fracture faces of plant material. Roots of Lemna minor are encapsulated in different nonpenetrating polymeric cryoprotectants, rapidly cooled in melting nitrogen and transferred to the pre-cooled cold stage of the AMR Biochamber. The technique has been used to follow the course of development of the phloem tissue in the root tip. These studies have shown that the phloem parenchyma appears to develop in a regular sequence. Unetched surfaces are virtually featureless, and it is necessary to remove a surface layer of water in order to visualize the biological structure. Themore » amount of water sublimed from the fractured surface is a function of both the time of etching and the water binding capacity of the cell contents. It is not possible to etch cells infiltrated with a penetrating cryoprotectant as the glycerol-water eutectic is stable at low temperatures and no water is lost from the fractured surface. Several distinct stages have been observed during the etching process.« less

Fiber reinforced solids possessing great fracture toughness: The role of interfacial strength

NASA Technical Reports Server (NTRS)

Atkins, A. G.

1975-01-01

The results of angle-ply investigations for strength and toughness of brittle fiber/brittle filament composites are presented. General results are discussed for both unidirectional and angle-ply intermittently bonded boron/epoxy composites as affected by soaking and freezing water environments. A description of and the operating instructions are included for the modified 230 mm (9 inch) wide intermittent coating tape making apparatus.

Cryovolcanic Conduit Evolution and Eruption on Icy Satellites

NASA Astrophysics Data System (ADS)

Mitchell, K. L.

2014-12-01

In silicate volcanism, such as on Earth or Io, eruptions typically result from fracture formation caused by interaction of tectonic stresses with inflating, pressurized magma sources, leading to transport of melt through an evolving conduit. On icy satellites the paradigm may be similar, resulting from some combination of tidal stresses and expansion of freezing water within, or near the base of, an ice shell. Such a fracture will result in eruption if mass continuity can be established, with buoyancy aided by exsolution and expansion of dissolved volatiles. After onset, conduit shape evolves due to: (1) shear-stresses or frictional erosional; (2) wallrock "bursting" due to massive wall stresses; (3) wall melting or condensation of particles due to heat transfer; or (4) changes in applied stresses. Preliminary thermodynamic and fluid mechanical analysis suggests some initial cooling during ascent resulting from exsolution and expansion of volatiles, thermally buffered by freezing, Conduit contraction may occur, and so evolution towards a deep, gas-filled plume chamber is difficult to accommodate without evoking a co-incidental process. Conduit flaring occurs near the surface where velocities are greatest, enhancing erosion. Here, viscous dissipative heating exceeds adiabatic cooling, and so some boiling (a few wt%) may occur. In contrast with silicate volcanism, decompression to below the triple point will occur within conduit, vent or jet, resulting in rapid freezing and boiling of the remaining water at a 6.8:1 ratio. Subsequent isentropic or adiabatic expansion within erupting jets may result in a few percent net of condensation or sublimation. These effects combined lead to ~4:1-7:1 solid:vapor ratios in the jet for most eruption conditions. These figures are consistent with the ~6:1 inferred in Enceladus' jets, supporting the hypothesis that the Enceladus plume draws from a subsurface body of liquids through a conduit. Similar results are anticipated if cryovolcanic plumes are confirmed on Europa. However, eruption from an ocean is more challenging there, as far greater volatile contents would be required to facilitate mass continuity from an initial fracture, and hence eruption, from the higher pressure source. Shallower sources, such as proposed under chaos, are less challenging.

Gap junctions contain different amounts of cholesterol which undergo unique sequestering processes during fiber cell differentiation in the embryonic chicken lens.

PubMed

Biswas, Sondip K; Lo, Woo-Kuen

2007-03-09

To determine the possible changes in the distribution of cholesterol in gap junction plaques during fiber cell differentiation and maturation in the embryonic chicken lens. The possible mechanism by which cholesterol is removed from gap junction plaques is also investigated. Filipin cytochemistry in conjunction with freeze-fracture TEM was used to visualize cholesterol, as represented by filipin-cholesterol complexes (FCCs) in gap junction plaques. Quantitative analysis on the heterogeneous distribution of cholesterol in gap junction plaques was conducted from outer and inner cortical regions. A novel technique combining filipin cytochemistry with freeze-fracture replica immunogold labeling (FRIL) was used to label Cx45.6 and Cx56 antibodies in cholesterol-containing gap junctions. Filipin cytochemistry and freeze-fracture TEM and thin-section TEM were used to examine the appearance and nature of the cholesterol-containing vesicular structures associated with gap junction plaques. Chicken lens fibers contain cholesterol-rich, cholesterol-intermediate and cholesterol-free gap junction populations in both outer and inner cortical regions. Filipin cytochemistry and FRIL studies confirmed that cholesterol-containing junctions were gap junctions. Quantitative analysis showed that approximately 86% of gap junctions in the outer cortical zone were cholesterol-rich gap junctions, whereas approximately 81% of gap junctions in the inner cortical zone were cholesterol-free gap junctions. A number of pleiomorphic cholesterol-rich vesicles of varying sizes were often observed in the gap junction plaques. They appear to be involved in the removal of cholesterol from gap junction plaques through endocytosis. Gap junctions in the young fibers are enriched with cholesterol because they are assembled in the unique cholesterol-rich cell membranes in the lens. A majority of cholesterol-rich gap junctions in the outer young fibers are transformed into cholesterol-free ones in the inner mature fibers during fiber cell maturation. A distinct endocytotic process appears to be involved in removing cholesterol from the cholesterol-containing gap junctions, and it may play a major role in the transformation of cholesterol-rich gap junctions into cholesterol-free ones during fiber cell maturation.

Adhesion at the interface in cured graphite fiber epoxy-amine resin composites

NASA Technical Reports Server (NTRS)

Needles, Howard L.; Alger, Kenneth W.; Okamoto, Robert

1987-01-01

The effect of high temperature curing on the interface between unsized or epoxy-sized graphite fiber tow and epoxy-amine resin was examined by scanning electron microscopy of compression and freeze fractured specimens. Little or no adhesion was found between the unsized graphite fiber tows and the epoxy-amine resin on curing at 165 C for 17 hrs. Epoxy-sized graphite fibers showed a similar lack of adhesion between the fiber tows and the epoxy-amine resin at 3 and 17 hr cures, although good penetration of the resin into the sized fiber tows had occurred. Interfacial bond strengths for the composites could not be effectively measured by compression fracture of specimens.

Spiderweb-like Fractures in Occator Crater

NASA Image and Video Library

2018-01-30

This image shows a complex set of fractures found in the southwestern region of the floor of Occator Crater on Ceres. In this picture, north is at the top. The two intersecting fracture systems (roughly northwest-southeast and southwest-northeast) are part of a larger fault network that extends across Occator's floor. These fractures have been interpreted as evidence that material came up from below and formed a dome shape, as if a piston was pushing Occator's floor from beneath the surface. This may be due to the upwelling of material coming from Ceres' deep interior. An alternative hypothesis is that the deformation is due to volume changes inside a reservoir of icy magma in the shallow subsurface that is in the process of freezing, similar to the change in volume that a bottle of water experiences when put in a freezer. Another set of fractures can be seen parallel to the southwestern wall and is not connected to the Occator fracture network. Dawn took this image during its extended mission on August 17, 2016, from its low-altitude mapping orbit, at a distance of about 240 miles (385 kilometers) above the surface. The image resolution is 120 feet (35 meters) per pixel. The center coordinates are 16 degrees north in latitude and 237 east in longitude. https://photojournal.jpl.nasa.gov/catalog/PIA22091

Nitrogen dioxide-induced alterations in ganglioside content and structure of pulmonary artery endothelial cell plasma membranes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sekharam, M.; Patel, J.M.; Block, E.R.

1990-02-26

Nitrogen dioxide (NO{sub 2}), an environmental oxidant, is known to cause injury to the surface of pulmonary artery endothelial cells (PAEC). Because gangliosides are present in the outer leaflet of plasma membranes, the authors hypothesize that NO{sub 2} exposure may alter the ganglioside content and structure of PAEC plasma membranes. To test this, confluent porcine PAEC were exposed to 5 ppm NO{sub 2} containing 5% CO{sub 2} for 48 hours at 37 C in a CO{sub 2} incubator. Controls were exposed to air containing 5% Co{sub 2} under identical conditions. After exposure: (1) total lipids were extracted and ganglioside basesmore » were separated and estimated by fluorescamine, (2) the sialic acid content of intact cells was measured by the resorcinol method, and (3) freeze-fracture analysis of the intact cell plasma membrane was done by propane jet freezing and shadowing with platinum and carbon to form a replica. The ganglioside and sialic acid/{mu}g protein, respectively. In No{sub 2}-exposed cells, ganglioside content was reduced by 45% and sialic acid content was increased by 30%. Freeze-fracture analysis of the plasma membrane of control cells showed the presence of 160{+-}12 particles/cm area at 45000x. In contrast, the number of particles on the No{sub 2}-exposed plasma membrane was reduced to 68{+-}5 particles/cm at 45000x (p < 0.05). These results demonstrate that NO{sub 2} causes structural changes in the surface of PAEC plasma membranes, and these are temporally associated with a reduction in the number of gagliosides in these cells.« less

Visualization of pores (export sites) correlated with cellulose production in the envelope of the gram-negative bacterium Acetobacter xylinum.

PubMed

Zaar, K

1979-03-01

The Gram-negative bacterium Acetobacter xylinum assembles a cellulse ribbon composed of a number of microfibrils in the longitudinal axis of its envelope. The zone of ribbon assembly was investigated by freeze-etch electron microscopy. Freeze-etching revealed, beneath the cellulose ribbons, a linear array of pores on the lipopolysaccharide membrane. These pores have a rim diameter of 120--150 A and a central hole or deepening of approximately 35 A. The axes of pore arrays closely coincide with linear arrays of 100 A particles on the E- and P-faces of the fractured lipopolysaccharide membranes. Pores and particles in the lipopolysaccharide membrane are probably congruent. The pores are hypothesized to be the export sites (penetration sites) for cellulose.

Cryo-scanning electron microscopy discloses differences in dehydration of frozen boar semen stored in large containers.

PubMed

Ekwall, H

2009-02-01

In general, freezing in flat plastic polyethylene terephthalate (PET) bags (FlatPacks) at 50 degrees C/min gives better post-thaw viability, in terms of sperm motility and membrane integrity, than does freezing in plastic maxi-straws, probably owing to differences in cryobiology. To test the hypothesis that this better survival post-thaw relates to the degree of sperm dehydration during freezing, the present study investigated the structure of boar semen in a frozen state using cryo-scanning electron microscopy (cryo-SEM) to compare two different packages (FlatPacks and maxi-straws) for single artificial insemination (AI) doses, and three different freezing rates. The semen was split-sample frozen in maxi-straws or FlatPacks (both holding 5 ml) using 3% glycerol as cryoprotectant. Three freezing rates were applied from -5 degrees C to -100 degrees C, namely 2 degrees C/min, 50 degrees C/min and 1200 degrees C/min, the lattermost by plunging the samples into liquid nitrogen (LN(2)). The samples were thereafter fractured into LN(2) and larger areas of extra-cellular, unbound frozen water ('ice lakes') were measured to determine the degree of dehydration of the spermatozoa. These areas decreased in size with an increase in cooling rate, the differences in size being more dramatic for maxi-straws than for FlatPacks. Size of ice lakes was also influenced by location within package in relation to cooling rate, the central values being always smaller in maxi-straws than in Flatpacks (p < 0.05 at 2 degrees C/min and 50 degrees C/min) but not at 1200 degrees C/min, which suggested the FlatPack allows for more homogenous freezing of boar semen.

Quantifying irreversible movement in steep, fractured bedrock permafrost on Matterhorn (CH)

NASA Astrophysics Data System (ADS)

Weber, Samuel; Beutel, Jan; Faillettaz, Jérome; Hasler, Andreas; Krautblatter, Michael; Vieli, Andreas

2017-02-01

Understanding rock slope kinematics in steep, fractured bedrock permafrost is a challenging task. Recent laboratory studies have provided enhanced understanding of rock fatigue and fracturing in cold environments but were not successfully confirmed by field studies. This study presents a unique time series of fracture kinematics, rock temperatures and environmental conditions at 3500 m a. s. l. on the steep, strongly fractured Hörnligrat of the Matterhorn (Swiss Alps). Thanks to 8 years of continuous data, the longer-term evolution of fracture kinematics in permafrost can be analyzed with an unprecedented level of detail. Evidence for common trends in spatiotemporal pattern of fracture kinematics could be found: a partly reversible seasonal movement can be observed at all locations, with variable amplitudes. In the wider context of rock slope stability assessment, we propose separating reversible (elastic) components of fracture kinematics, caused by thermoelastic strains, from the irreversible (plastic) component due to other processes. A regression analysis between temperature and fracture displacement shows that all instrumented fractures exhibit reversible displacements that dominate fracture kinematics in winter. Furthermore, removing this reversible component from the observed displacement enables us to quantify the irreversible component. From this, a new metric - termed index of irreversibility - is proposed to quantify relative irreversibility of fracture kinematics. This new index can identify periods when fracture displacements are dominated by irreversible processes. For many sensors, irreversible enhanced fracture displacement is observed in summer and its initiation coincides with the onset of positive rock temperatures. This likely indicates thawing-related processes, such as meltwater percolation into fractures, as a forcing mechanism for irreversible displacements. For a few instrumented fractures, irreversible displacements were found at the onset of the freezing period, suggesting that cryogenic processes act as a driving factor through increasing ice pressure. The proposed analysis provides a tool for investigating and better understanding processes related to irreversible kinematics.

Preparation of cryofixed cells for improved 3D ultrastructure with scanning transmission electron tomography.

PubMed

Höhn, Katharina; Sailer, Michaela; Wang, Li; Lorenz, Myriam; Schneider, Marion E; Walther, Paul

2011-01-01

Scanning transmission electron tomography offers enhanced contrast compared to regular transmission electron microscopy, and thicker samples, up to 1 μm or more, can be analyzed, since the depth of focus and inelastic scattering are not limitations. In this study, we combine this novel imaging approach with state of the art specimen preparation by using novel light transparent sapphire specimen carrier for high-pressure freezing and a freeze substitution protocol for better contrast of membranes. This combination allows for imaging membranes and other subcellular structures with unsurpassed quality. This is demonstrated with mitochondria, where the inner and outer mitochondrial membranes as well as the membranes in the cristae appear in very close apposition with a minimal intermembrane space. These findings correspond well with old observations using freeze fracturing. In 880-nm thick sections of hemophagocytes, the three-dimensional structure of membrane sheets could be observed in the virtual sections of the tomogram. Microtubules, actin and intermediate filaments could be visualized within one sample. Intermediate filaments, however, could even be better observed in 3D using surface scanning electron tomography.

Cytoplasmic membrane changes during adaptation of the fresh water cyanobacterium Synechococcus 6311 to salinity

NASA Technical Reports Server (NTRS)

Lefort-Tran, M.; Pouphile, M.; Spath, S.; Packer, L.

1988-01-01

In this investigation, changes were characterized in cell structure and cytoplasmic membrane organization that occur when the freshwater cyanobacterium Synechococcus 6311 is transferred from 'low salt' (0.03 molar NaCl) to 'high salt' (0.5 molar NaCl) media (i.e. sea water concentration). Cells were examined at several time points after the imposition of the salt stress and compared to control cells, in thin sections and freeze fracture electron microscopy, and by flow cytometry. One minute after exposure to high salt, i.e. 'salt shock', virtually all intracellular granules disappeared, the density of the cytoplasm decreased, and the appearance of DNA material was changed. Glycogen and other granules, however, reappeared by 4 hours after salt exposure. The organization of the cytoplasmic membrane undergoes major reorganization following salt shock. Freeze-fracture electron microscopy showed that small intramembrane particles (diameter 7.5 and 8.5 nanometers) are reduced in number by two- to fivefold, whereas large particles, (diameters 14.5 and 17.5 nanometers) increase two- to fourfold in frequency, compared to control cells grown in low salt medium. The changes in particle size distribution suggest synthesis of new membrane proteins, in agreement with the known increases in respiration, cytochrome oxidase, and sodium proton exchange activity of the cytoplasmic membrane.

Imaging of single liver tumor cells intoxicated by heavy metals using ToF-SIMS

NASA Astrophysics Data System (ADS)

Mai, Fu-Der; Chen, Bo-Jung; Wu, Li-Chen; Li, Feng-Yin; Chen, Wen-Kang

2006-07-01

Human liver tumor cells intoxicated with five different Cd, Cu, Cr, Hg and Zn metals were analyzed using imaging time-of-flight secondary ion mass spectrometry (ToF-SIMS) to visualize the metal distributions in a single cell basis. A protocol was developed by combining rapid freezing, freeze-fracture and imprinting for transferring the intoxicated cells to a silicon wafer. As shown in the ToF-SIMS images, the cellular morphology was preserved indicating that this protocol can be used to prepare a representative cell for ToF-SIMS imaging analysis. Among the five metal ions investigated in this study, only Cr and Cu ions show preferential diffusion into the cell after simulated intoxication while the signals of the other three ions are either too low to be detected or unable to be distinguished from background intensity.

Animal-vegetal polarity in the plasma membrane of a molluscan egg: a quantitative freeze-fracture study.

PubMed

Speksnijder, J E; Mulder, M M; Dohmen, M R; Hage, W J; Bluemink, J G

1985-03-01

Using freeze-fracture electron microscopy, the numerical particle distribution in the fertilized Nassarius egg plasma membrane has been analyzed in four areas at different positions along the animal-vegetal axis of the egg. These areas can be distinguished by distinct microvilli patterns and differences in microvilli densities. In all areas, more IMPs (intramembrane particles) are present on the P face than on the corresponding E face. The ratio of the number of IMPs present on E and P face is similar in all areas (0.48-0.55) except for the most animal part of the vegetal hemisphere, where relatively more IMPs remain attached to the exterior half of the fractured membrane (E/P ratio = 0.88). The IMP density at the vegetal pole of the egg is considerably higher than in the animal hemisphere and in the animal part of the vegetal hemisphere. This difference is due to an increased number of IMPs in all size classes (4-18 nm). In the area adjacent to the vegetal pole the density of particles is also higher than in the two more animal areas, but here the difference is exclusively due to the smaller IMP size classes (4-8 nm). Statistical analysis of our data reveals that the area adjacent to the vegetal pole patch is significantly different from the other areas with respect to the distribution of the IMPs over the different IMP size classes. These results demonstrate the polar organization of the Nassarius egg plasma membrane. The possible role of this surface heterogeneity in the spatial organization of the egg cell and the later embryo is discussed.

Vasopressin Receptor Signaling and Cycling of Water Channels in Renal Epithelia (HBC)

DTIC Science & Technology

1993-08-27

in the cellular ultrastructure that results in mucosal membrane reorganization to numerous microvilli from the normal phase of microridges ( DiBona ...a restructuring of the apical membrane microridges into numerous microvilli with an increase in membrane surface area ( DiBona , 1981; DiBona et aL...markers in freeze-fracture studies of toad urinary bladder. J. Histochem. Cytochem., 35, 1405-1414. DiBona , D.R. 1981. Vasopressin action on the

Effects of proof loads and combined mode loadings on fracture and flaw growth characteristics of aerospace alloys

NASA Technical Reports Server (NTRS)

Shah, R. C.

1974-01-01

This experimental program was undertaken to determine the effects of (1) combined tensile and bending loadings, (2) combined tensile and shear loadings, and (3) proof overloads on fracture and flaw growth characteristics of aerospace alloys. Tests were performed on four alloys: 2219-T87 aluminum, 5Al-2.5Sn (ELl) titanium, 6Al-4V beta STA titanium and high strength 4340 steel. Tests were conducted in room air, gaseous nitrogen at -200F (144K), liquid nitrogen and liquid hydrogen. Flat center cracked and surface flawed specimens, cracked tube specimens, circumferentially notched round bar and surface flawed cylindrical specimens were tested. The three-dimensional photoelastic technique of stress freezing and slicing was used to determine stress intensity factors for surface flawed cylindrical specimens subjected to tension or torsion. Results showed that proof load/temperature histories used in the tests have a small beneficial effect or no effect on subsequent fracture strength and flaw growth rates.

Modeling Sea Ice Trajectories for Oil Spill Tracking.

DTIC Science & Technology

1981-06-01

is compared with sea ice motions observed during the AIDJEX main field experiment in the Beaufort Sea from April 1975 to February 1976. The average ...more recently grown on leads formed as the floes fracture and divide. The large-scale average thickness of the pack ice is roughly 3 m. As an...opposite extreme, during the summer when air temperatures rise above freezing, melting and offshore winds combine to form an approximately 300-km-wide swath

Summary of events and geotechnical factors leading to decommissioning of the Strategic Petroleum Reserve (SPR) facility at Weeks Island, Louisiana

DOE Office of Scientific and Technical Information (OSTI.GOV)

Neal, J.T.; Bauer, S.J.; Ehgartner, B.L.

1996-10-01

A sinkhole discovered over the edge of the Strategic Petroleum Reserve storage facility at Weeks Island salt dome, Louisiana, led to decommissioning the site during 1995--1998, following extensive diagnostics in 1994. The sinkhole resulted from mine-induced fractures in the salt which took may years to develop, eventually causing fresh water to leak into the storage chamber and dissolve the overlying salt, thus causing overburden collapse into the void. Prior to initiating the oil removal, a freeze wall was constructed at depth around the sinkhole in 1995 to prevent water inflow; a freeze plug will remain in place until the minemore » is backfilled with brine in 1997--8, and stability is reached. Residual oil will be removed; environmental monitoring has been initiated and will continue until the facility is completely plugged and abandoned, and environmental surety is achieved.« less

A comparison between rib fracture patterns in peri- and post-mortem compressive injury in a piglet model.

PubMed

Bradley, Amanda L; Swain, Michael V; Neil Waddell, J; Das, Raj; Athens, Josie; Kieser, Jules A

2014-05-01

Forensic biomechanics is increasingly being used to explain how observed injuries occur. We studied infant rib fractures from a biomechanical and morphological perspective using a porcine model. We used 24, 6th ribs of one day old domestic pigs Sus scrofa, divided into three groups, desiccated (representing post-mortem trauma), fresh ribs with intact periosteum (representing peri-mortem trauma) and those stored at -20°C. Two experiments were designed to study their biomechanical behaviour fracture morphology: ribs were axially compressed and subjected to four-point bending in an Instron 3339 fitted with custom jigs. Morphoscopic analysis of resultant fractures consisted of standard optical methods, micro-CT (μCT) and Scanning Electron Microscopy (SEM). During axial compression fresh ribs did not fracture because of energy absorption capabilities of their soft and fluidic components. In flexure tests, dry ribs showed typical elastic-brittle behaviour with long linear load-extension curves, followed by short non-linear elastic (hyperelastic) behaviour and brittle fracture. Fresh ribs showed initial linear-elastic behaviour, followed by strain softening and visco-plastic responses. During the course of loading, dry bone showed minimal observable damage prior to the onset of unstable fracture. Frozen then thawed bone showed similar patterns to fresh bone. Morphologically, fresh ribs showed extensive periosteal damage to the tensile surface with areas of collagen fibre pull-out along the tensile surface. While all dry ribs fractured precipitously, with associated fibre pull-out, the latter feature was absent in thawed ribs. Our study highlights the fact that under controlled loading, fresh piglet ribs (representing perimortem trauma) did not fracture through bone, but was associated with periosteal tearing. These results suggest firstly, that complete lateral rib fracture in infants may in fact not result from pure compression as has been previously assumed; and secondly, that freezing of bone during storage may affect its fracture behaviour. Copyright © 2013 Elsevier Ltd. All rights reserved.

Molecule-Specific Imaging Analysis of Carcinogens in Breast Cancer Cells Using Time-of-Flight Secondary Ion Mass Spectrometry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Quong, J N; Knize, M G; Kulp, K S

2003-08-19

Imaging time-of-flight secondary ion mass spectrometry (TOF-SIMS) is used to study the localization of heterocyclic amines in MCF7 line of human breast cancer cells. The detection sensitivities of a model rodent mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were determined. Following an established criteria for the determination of status of freeze-fracture cells, the distribution of PhIP in the MCF7 cells are reported.

Characterization of outer membranes isolated from Treponema pallidum, the syphilis spirochete.

PubMed

Radolf, J D; Robinson, E J; Bourell, K W; Akins, D R; Porcella, S F; Weigel, L M; Jones, J D; Norgard, M V

1995-11-01

Previous freeze-fracture electron microscopy (EM) studies have shown that the outer membrane (OM) of Treponema pallidum contains sparse transmembrane proteins. One strategy for molecular characterization of these rare OM proteins involves isolation of T. pallidum OMs. Here we describe a simple and extremely gentle method for OM isolation based upon isopycnic sucrose density gradient ultracentrifugation of treponemes following plasmolysis in 20% sucrose. Evidence that T. pallidum OMs were isolated included (i) the extremely low protein/lipid ratio of the putative OM fraction, (ii) a paucity of antigenic and/or biochemical markers for periplasmic, cytoplasmic membrane, and cytosolic compartments, and (iii) freeze-fracture EM demonstrating that the putative OMs contained intramembranous particles highly similar in size and density to those in native T. pallidum OMs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that the OMs contained a relatively small number of treponemal proteins, including several which did not appear to correspond to previously characterized T. pallidum antigens. Interestingly, these candidate rare OM proteins reacted poorly with syphilitic sera as determined by both conventional immunoblotting and enhanced chemiluminescence. Compared with whole cells, T. pallidum OMs were deficient in cardiolipin, the major lipoidal antigen reactive with antibodies in syphilitic sera. Also noteworthy was that other lipoidal constituents of OMs, including the recently discovered glycolipids, did not react with human syphilitic sera. These latter observations suggest that the poor antigenicity of virulent T. pallidum is a function of both the lipid composition and the low protein content of its OM.

Drying techniques for the visualisation of agarose-based chromatography media by scanning electron microscopy.

PubMed

Nweke, Mauryn C; Turmaine, Mark; McCartney, R Graham; Bracewell, Daniel G

2017-03-01

The drying of chromatography resins prior to scanning electron microscopy is critical to image resolution and hence understanding of the bead structure at sub-micron level. Achieving suitable drying conditions is especially important with agarose-based chromatography resins, as over-drying may cause artefact formation, bead damage and alterations to ultrastructural properties; and under-drying does not provide sufficient resolution for visualization under SEM. This paper compares and contrasts the effects of two drying techniques, critical point drying and freeze drying, on the morphology of two agarose based resins (MabSelect™/d w ≈85 µm and Capto™ Adhere/d w ≈75 µm) and provides a complete method for both. The results show that critical point drying provides better drying and subsequently clearer ultrastructural visualization of both resins under SEM. Under this protocol both the polymer fibers (thickness ≈20 nm) and the pore sizes (diameter ≈100 nm) are clearly visible. Freeze drying is shown to cause bead damage to both resins, but to different extents. MabSelect resin encounters extensive bead fragmentation, whilst Capto Adhere resin undergoes partial bead disintegration, corresponding with the greater extent of agarose crosslinking and strength of this resin. While freeze drying appears to be the less favorable option for ultrastructural visualization of chromatography resin, it should be noted that the extent of fracturing caused by the freeze drying process may provide some insight into the mechanical properties of agarose-based chromatography media. © 2017 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hypersolidus geothermal energy from the moving freeze-fracture-flow boundary

NASA Astrophysics Data System (ADS)

Carrigan, Charles; Eichelberger, John; Sigmundsson, Freysteinn; Papale, Paolo; Sun, Yunwei

2014-05-01

Rhyolitic magmas at low pressure undergo much of their crystallization over a small temperature interval just above the solidus. This hypersolidus material has a high energy density and effective heat capacity because of stored heat of crystallization, yet may sustain fractures and therefore admit heat exchange with fluids because of its interlocking crystal framework. Rhyolitic magmas emplaced near the liquidus should at first cool rapidly, owing to internal convection, modest crystallization with declining temperature, and extreme temperature gradients at their boundaries. However, once the solidus is approached the rapid rise in effective heat capacity should result in low temperature gradients and rates of heat flow within the bodies. They are suspended for a time in the hypersolidus state. Prodigious quantities of heat can be released from these thermal masses by hydrothermal systems, natural or perhaps stimulated, fracturing their way inward from the margins. The fracture front drives the solidus isotherm ahead of it. Heat of crystallization in front of the advancing solidus is transferred across the thin, moving boundary zone to the external fluid, which advects it away. Once the material is below (outboard of) the solidus, it behaves as normal rock and cools rapidly, having a heat capacity only about 20% that of water. Variations on this theme were published by Lister (1974) for mid-ocean ridges, Hardee (1980) for lava lakes, and Bjornsson et al (1982) for Grimsvotn and Heimaey, who cited possible geothermal energy exploitiation. This scenario is consistent with a number of observations: 1. The geophysical rarity of imaging mostly liquid magma in the shallow crust, despite common petrologic evidence that silicic magma has undergone shallow storage. 2. More common imaging of "partial melt" volumes, whose inferred properties suggest some, but not dominant proportion of melt. 3. Evidence that pure-melt rhyolitic eruptions may have drained relatively shallow hypersolidus plutons. 4. Downward propagating thin conductive boundary zone observed in repeated coring of Kilauea Iki lava lake, Hawaii 5. Record enthalpy flow and temperature during flow-testing of Iceland Deep Drilling Project (IDDP)-1 in Krafla Caldera by Landsvirkjun Co. Production came from a 2.1-km-deep 500oC "magma" contact zone, from the vicinity of which fresh rhyolite glass-bearing felsite and crystal-poor rhyolite glass fragments were recovered. The hypothesis of a moving freeze-fracture-flow boundary raises the possibility of ultra-high-temperature, natural or engineered geothermal systems in volcanic areas. We believe that this prospect, as well as the benefit to understanding volcanic hazards at restless calderas, gives merit to further exploration of the hypersolidus regime beneath Krafla Caldera.

Investigation on the modification behavior of A356.2 alloy with Yb-La composite modifier

NASA Astrophysics Data System (ADS)

Zhang, Shaochen; Leng, Jinfeng; Wang, Zhibin; Wang, Qi; Shao, Yuewen; Teng, Xinying

2018-01-01

In this work, an investigation was conducted to evaluate the influence of the Yb + La addition on the microstructures and mechanical properties of A356.2 alloy. The results show that the the grain size was refined and morphological structure of Si changed from needle-like to fine spheroidal particle by adding Yb + La. The eutectic temperature decreased from 589 °C to 581 °C and the freezing range of primary α-Al become wider after adding Yb + La. The mechanical properties showed great improvement after T6 heat treatment. The ultimate tensile strength and microhardness of the composites were increased to 279.0 MPa and 99.2 HB after T6 heat treatment which increased by 28.2% and 47.3%, respectively. The fracture mechanism was analyzed using the scanning electron microscopy and proved to be a transgranular/intergranular fracture modes.

A SIMPLE FREEZE-FRACTURE REPLICATION METHOD FOR ELECTRON MICROSCOPY

PubMed Central

Bullivant, Stanley; Ames, Adelbert

1966-01-01

A simple method to achieve results similar to the freeze-etching technique of Moor et al. (1961) is described. The frozen tissue is cut under liquid nitrogen with a razor blade outside the evaporator rather than inside with a cooled microtome. The conditions of the experiment do not favor sublimation, and it is proposed that the structure of the replica be explained by local faults in the cleavage plane which leaves structures, such as membranes, standing above the ice. Micrographs of replicas of glycerol-protected frozen small intestine of mouse prepared by the method are presented and the structural details they show are discussed. The problem of vapor-deposited contamination is discussed. It is concluded that this is a practical method for obtaining electron micrographs that are relatively free of artifact, and that further improvements may be expected from the use of rapidly frozen fresh tissue and a clean vacuum system, possibly of the ion-pumped type. PMID:5962938

Self-assembly of chlorophenols in water

PubMed Central

Rogalska, Ewa; Rogalski, Marek; Gulik-Krzywicki, Tadeusz; Gulik, Annette; Chipot, Christophe

1999-01-01

In saturated solutions of some di- and trichlorophenols, structures with complex morphologies, consisting of thin, transparent sheets often coiling into helices and ultimately twisting into filaments, were observed under the optical microscope. Freeze-fracture electron microscopy, x-ray diffraction, phase diagrams, and molecular modeling were performed to elucidate the observed phenomena. Here, we present evidence that the chlorophenols studied, when interacting with water, self-assemble into bilayers. The fact that some chlorophenols form the same supramolecular structures as those described previously for structurally nonrelated surfactants sheds light on the mechanisms of self-assembly. PMID:10359753

Management of unicameral bone cyst by using freeze dried radiation sterilized bone allograft impregnate with autogenous bone marrow.

PubMed

Datta, N K; Das, K P; Alam, M S; Kaiser, M S

2014-07-01

Unicameral bone cyst is a common benign bone tumor and most frequent cause of the pathological fracture in children. We have started a prospective study for that treatment of unicameral bone cyst by using freeze dried radiation sterilized bone allograft impregnated with autogenous bone marrow in the department of Orthopaedics, Bangabandhu Sheikh Mujib Medical University (BSMMU) during May 1999 to April 2012. Aim of this study was to see Freeze dried radiation sterilized bone allograft impregnate with autogenous bone marrow a satisfactory graft material in the treatment of unicameral bone cyst as well as factors such as patients age, sex, cyst size and site of lesion influence on cyst healing. A total 35 patients of unicameral bone cyst were operated. In this study out of 35 patients, male were 22(62.86%) and female were 13(37.14). Male Female ratio 22:13(1.70:1) Age of the patients ranging from 2 years 6 month to 20 years, mean age 12.18 years more common 11 years to 20 years 29(82.86%) patients. Common bones sites involvements are proximal end of Humerus 20(57.14%), proximal end of Femur 7(20 %), proximal end of Tibia 3(8.57%), Calcanium 2(5.71%), proximal end of Ulna 1(2.86%), shaft of Radius 1(2.86%) and Phalanx 1(2.86%). Final clinical outcome of unicameral bone cyst treated by thorough curettage of cavity and tightly filled with freeze dried radiation sterilized bone allograft impregnate with autogenous bone marrow in which healed (success rate) 88.57% (31) and recurrence rate is 11.43% (4). P value is <0.001. Follow up period was 6 month to 11 years. From our study it was realized that freeze dried radiation sterilized bone allograft impregnated with autogenous bone marrow is useful graft material for healing of the lesional area as well as restoring structural integrity for the treatment of unicameral bone cyst.

Thermal regimes in bedrock and open fractures in the Nordnes rockslide, Norway

NASA Astrophysics Data System (ADS)

Hvidtfeldt Christiansen, Hanne; Harald Blikra, Lars

2010-05-01

The Nordnes rockslide site is located in the arctic part of the periglacial mountain landscape of Northern Norway at 69°30'N. It consists in the upper part of 1-10 m wide and 1-10 m deep open fractures. Extensive displacements measurements using GPS surveys, crackmeters, tiltmeter and lasers establish the intermunicipality monitoring programme, which shows ongoing deformation of the rockslide. In the rather special topographical setting of the open fractures we have during the International Polar Year 2007 to 2009 recorded the thermal regime of the upper part of the bedrock and of the air in the cracks for attempting to determine whether the recorded deformation can be geomorphologically controlled by bedrock surface expansion and contraction and/or by seasonal freezing or even by permafrost, or if only normal gravitational processes control the observed displacements. The upper 40 cm bedrock thermal conditions have been investigated in different exposures to identify the seasonal freezing depth and length, for determination of the influence of potential ice segregation processes causing weathering of the bedrock surfaces. The data show generally that that the bedrock surface is in the -3 to -8C freezing window for 3 to 6 months. Likewise 250 cm deep bedrock thermal monitoring have been carried out in three boreholes during one year at 900 m, 800 m and 625 m asl. extending over the area from the upper part of the unstable area and into the stable area above, for determination of the regional permafrost zone. These results in combination with thermal evidence from other deeper boreholes from the same setting in the same region show that seasonal freezing extends 5-10 m down, and that a potential active layer also is in the order of 5-10 m deep. The air temperatures in the cracks show significant cooling during winter, when the cracks have a thick snow cover, thus demonstrating the potential existence of permafrost in deeper part of the cracks and in the ground just around these. Automatic photography has been used for the last 4 years to study the seasonal snow cover duration and thickness in the open cracks. This shows that a thicker snow cover only develops in mid winter, with maximum amounts of snow in March and April, but also that not all snow melts during summer in the deeper parts of the open cracks. In addition we have found small pockets of ice in closed spaces of the bottom parts of the open crack, indicating the presence of permafrost. The combination of thermal data and the special seasonal variation in the rockslide deformations indicate that most likely refreezing of snow meltwater goes on in the open cracks for a considerable period from late summer, autumn and into the early winter, when the recorded rockslide deformation is largest. In late winter no significant deformation is recorded when the ground is constantly frozen, but there is a significant potential for ice segregation to occur where moisture is present in the rock.

In vitro early changes in intercellular junctions by treatment with a chemical carcinogen.

PubMed

Tachikawa, T; Kohno, Y; Matsui, Y; Yoshiki, S

1986-06-01

To examine early intercellular junction changes caused by treatment with 9,10-dimethyl-1,2-benzanthracene (DMBA), rat lingual epithelium was cultivated in isolation and observed by electrophysiological, freeze-fracture and whole-mount electron microscopy. Electrophysiological measurements showed a transient decrease in membrane potential of -10.2 mV 6 h after the treatment. It returned to almost the same level as that of the control group 1 day later. Six hours after treatment, input resistance decreased rapidly to 5.3 M omega but increased to 18.0 M omega 12 h after treatment. Transient reduction of input resistance and membrane potential occurred prior to the decrease in the coupling ratio 6 h after treatment with DMBA. In freeze-fracture replicas, the number of gap junctions decreased by approximately 45% of the control value 6 h after treatment with DMBA. At 12 h and thereafter, the number and area of gap junctions subsequently decreased by 60-80% of the control value. Alterations in the number and area of desmosomes were similar to those of the gap junctions. The formation of epithelial cytoskeletons, partially devoid of the 2-4 and 5-8 nm filaments was also observed. A decrease in the density of filament networks beneath the plasma membranes was especially apparent. Treatment with a carcinogen brought about morphological cellular changes as early as 6 h after treatment, and such early changes might trigger metabolic cellular abnormalities. Affected cells appear to move away from normal cells in a process of repeated destruction and revision of intercellular junctions, and cytoskeletons.

Cold-induced ultrastructural changes in bull and boar sperm plasma membranes.

PubMed

De Leeuw, F E; Chen, H C; Colenbrander, B; Verkleij, A J

1990-04-01

The effect of low temperatures on the ultrastructure of the plasma membrane of bull and boar spermatozoa was investigated. Cold-induced changes in the organization of sperm plasma membrane components were demonstrated by the use of fast-freezing combined with freeze-fracture electron microscopy. This preparation technique ensures fixation without artifacts. At 38 degrees C bull and boar spermatozoa exhibited a random distribution of intramembranous particles over the plasma membrane of both head and tail. Exposure to 0 degree C resulted in redistribution of the intramembranous particles: on the head and principal piece of bull spermatozoa and on the principal piece of boar spermatozoa, particle-free areas were observed, whereas on the boar sperm head, particle aggregates were present. The original particle distribution was restored upon rewarming of bull and boar spermatozoa to 38 degrees C, as well as after freezing and thawing of bull spermatozoa. Dilution of bull and boar semen into Tris-dilution buffer and Beltsville Thaw Solution-dilution buffer, respectively, could not prevent cold-induced redistribution of intramembranous particles. The observed particle reorganization upon cooling was interpreted as the result of lateral phase separation in the plasma membrane. Species-dependent differences in cold-induced ultrastructural changes were considered to be determined by lipid composition and asymmetry of the plasma membrane, and might be related to differences in cold resistance between species.

Molecular imaging of lipids in cells and tissues

NASA Astrophysics Data System (ADS)

Borner, Katrin; Malmberg, Per; Mansson, Jan-Eric; Nygren, Hakan

2007-02-01

The distribution pattern of lipid species in biological tissues was analyzed with imaging mass spectrometry (TOF-SIMS; time-of-flight secondary ion mass spectrometry). The first application shows distribution of a glycosphingolipid, the galactosylceramide-sulfate (sulfatide) with different hydrocarbon chain lengths and the fatty acids palmitate and oleate in rat cerebellum. Sulfatides were seen localized in regions suggested as paranodal areas of rat cerebellar white matter as well as in the granular layer, with highest concentrations at the borders of the white matter. Different distribution patterns could be shown for the fatty acid C16:0 palmitate and C18:1 oleate in rat cerebellum, which seem to origin partly from the hydrocarbon chains of phosphatidylcholine. Results were shown for two different tissue preparation methods, which were plunge-freezing and cryostat sectioning as well as high-pressure freezing, freeze-fracturing and freeze-drying. The second application shows TOF-SIMS analysis on a biological trial of choleratoxin treatment in mouse intestine. The effect of cholera toxin on lipids in the intestinal epithelium was shown by comparing control and cholera toxin treated mouse intestine samples. A significant increase of the cholesterol concentration was seen after treatment. Cholesterol was mainly localized to the brush border of enterocytes of the intestinal villi, which could be explained by the presence of cholesterol-rich lipid rafts present on the microvilli or by relations to cholesterol uptake. After cholera toxin exposure, cholesterol was seen increased in the nuclei of enterocytes and apparently in the interstitium of the villi. We find that imaging TOF-SIMS is a powerful tool for studies of lipid distributions in cells and tissues, enabling the elucidation of their role in cell function and biology.

Comparative study of subtalar arthrodesis after calcaneal frature malunion with autologous bone graft or freeze-dried xenograft.

PubMed

Henning, Carlo; Poglia, Gabriel; Leie, Murilo Anderson; Galia, Carlos Roberto

2015-12-01

Calcaneal fracture malunion may evolve into arthrosis and severe foot deformities. The aim of this study was to identify differences in bony union following corrective subtalar arthrodesis with interposition of autologous tricortical bone graft or freeze-dried bovine xenograft. We prospectively evaluated 12 patients who underwent subtalar arthrodesis, six patients received autografts and 6 received freeze-dried bovine xenografts. After a mean followup of 58 weeks, the patients were clinical assessed using AOFAS scale and the visual analog scale (VAS) for pain and for final radiographic parameters measurement. Two blind raters evaluated the length of time required for solid union of the arthrodesis and graft integration by retrospective radiographic examination. In the autograft group, AOFAS score improved from a preoperative average of 37 to 64 points postoperatively (p = 0.02) and mean VAS score improved from 4.7 to 1.9 (p = 0.028). In the xenograft group, AOFAS score improved from 38 to 74 points (p = 0.02) and VAS from 5.5 to 2.7 (p = 0.046). Solid union was achieved in all cases in the autograft group at an average of 5.3 weeks and in five cases in the xenograft group at 8.8 weeks (p = 0.077). Graft integration occurred after an average of 10.7 weeks in the autograft group and 28.8 weeks in the xenograft group (p = 0.016). With the numbers available, no significant difference could be detected in the length of time required for solid union of subtalar arthrodesis between groups, although time to integration of freeze-dried bovine xenografts was statistically higher. Clinical and functional improvement was observed in both groups.

An ultrastructural analysis of the epithelial-fiber interface (EFI) in primate lenses.

PubMed

Kuszak, J R; Novak, L A; Brown, H G

1995-11-01

The purpose of this study was to conduct a comprehensive ultrastructural analysis of the epithelial-fiber interface (EFI) in normal adult primate (Macaque nemestrina and fascicularis; 6-9 years old, n = 10) lenses. Scanning electron microscopy (SEM) was used to initially characterize the gross size, shape and three-dimensional organization of central zone (cz) epithelial cells and the anterior ends of elongating fibers beneath these cells. This fiducial information was essential to properly orient lens pieces in freeze fracture specimen carriers for the production of replicas with unambiguously identifiable EFI. Transmission electron microscopy (TEM) of replicas and thin-sectioned material were used to ultrastructurally analyse the cz EFI. TEM thin-sectioned material was also used to ultrastructurally analyse the pregerminative (pgz), germinative (gz) and transitional zone (tz) EFI. Correlative SEM and TEM of cz EFI components revealed that the apical membrane of both epithelial and elongating fiber cells were irregularly polygonal in shape, and aligned in parallel as smooth, concave-convex surfaces. However, whereas epithelial cell apical surfaces had minimal size variation, elongating fibers were larger and considerably variable in size. Quantitative analysis of > 10000 micron2 cz elongating fiber apical surfaces failed to detect any gap junctions defined in freeze fracture replicas as complementary aggregates of transmembrane proteins (connexons) conjoined across a narrowed extracellular space. However, a comparable frequency of vesicular events was noted in this region as quantified previously in adult and embryonic chick lens. Correlative TEM analysis > 1500 linear micrometers of thin-sectioned EFI from this region confirmed the presence of epithelial-epithelial gap junctions, elongating fiber-elongating fiber gap junctions, and an extreme paucity of epithelial-elongating fiber gap junctions. In contrast, TEM analysis of > 1000 linear micrometers of thin-sectioned pgz, gz and tz EFI, confirmed the presence of epithelial-epithelial gap junctions, elongating fiber-elongating fiber gap junctions, numerous epithelial-elongating fiber adherens junctions and a few epithelial-elongating fiber gap junctions. Thus, the results of this and previous quantitative morphological and physiological studies (electronic and dye coupling) demonstrate that there is limited coupling between cz epithelial cells and underlying elongating fibers. Furthermore, the absence of gap junctional plaques in cz EFI freeze-fracture replicas and either pentalaminar or septalaminar profiles in correlative thin-sections, suggests that this limited coupling could be mediated via isolated gap junction channels. However, the results of this and previous quantitative studies further show that a greater degree of coupling exists across the pgz, gz and tz regions of the EFI and that this coupling is likely to be mediated by gap junction plaques. Finally, this and other studies continue to demonstrate that transcytotic processes play a role in lens physiology at the EFI.

Relative intranuclear magnesium and phosphorus contents in normal and tumor cells of the human thyroid gland as revealed by energy-dispersive X-ray microanalysis.

PubMed

Lukács, G L; Zs-Nagy, I; Steiber, J; Györi, F; Balázs, G

1996-01-01

Energy dispersive X-ray microanalysis was performed on altogether 42 surgically removed tissue specimens of 32 patients, which were taken either from intact thyroid parts or various histopathologically verified tumors of the thyroid gland. The tissue specimens were processed with the freeze-fracture-freeze-drying technique and then analyzed in the so-called bulk specimen form. The studies were carried out during the years 1980-81, when intranuclear monovalent ionic composition was studied in detail. From the retained total elemental peak list, it was possible to calculate retrospectively the relative intranuclear Mg and P contents. The data processed by nested (hierarchical) analysis of variance show that the intranuclear Mg content of the 5 diagnostic groups (normal thyroid tissue, thyroiditis, benign adenomas, differentiated carcinomas and undifferentiated thyroid tumors) increases significantly, in parallel with the increasing malignancy, but the P content remains unchanged. One can conclude that the elevated intranuclear Mg content in the tumors of high malignancy may be of diagnostic importance, and a warning signal for the therapeutic approaches based on Mg-supplementations.

Moisture migration, microstructure damage and protein structure changes in porcine longissimus muscle as influenced by multiple freeze-thaw cycles.

PubMed

Zhang, Mingcheng; Li, Fangfei; Diao, Xinping; Kong, Baohua; Xia, Xiufang

2017-11-01

This study investigated the effects of multiple freeze-thaw (F-T) cycles on water mobility, microstructure damage and protein structure changes in porcine longissimus muscle. The transverse relaxation time T 2 increased significantly when muscles were subjected to multiple F-T cycles (P<0.05), which means that immobile water shifted to free water and the free water mobility increased. Multiple F-T cycles caused sarcomere shortening, Z line fractures, and I band weakening and also led to microstructural destruction of muscle tissue. The decreased free amino group content and increased dityrosine in myofibrillar protein (MP) revealed that multiple F-T cycles caused protein cross-linking and oxidation. In addition, the results of size exclusion chromatography, circular dichroism spectra, UV absorption spectra, and intrinsic fluorescence spectroscopy indirectly proved that multiple F-T cycles could cause protein aggregation and degradation, α-helix structure disruption, hydrophobic domain exposure, and conformational changes of MP. Overall, repeated F-T cycles changed the protein structure and water distribution within meat. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cellular control over spicule formation in sea urchin embryos: A structural approach.

PubMed

Beniash, E; Addadi, L; Weiner, S

1999-03-01

The spicules of the sea urchin embryo form in intracellular membrane-delineated compartments. Each spicule is composed of a single crystal of calcite and amorphous calcium carbonate. The latter transforms with time into calcite by overgrowth of the preexisting crystal. Relationships between the membrane surrounding the spiculogenic compartment and the spicule mineral phase were studied in the transmission electron microscope (TEM) using freeze-fracture. In all the replicas observed the spicules were tightly surrounded by the membrane. Furthermore, a variety of structures that are related to the material exchange process across the membrane were observed. The spiculogenic cells were separated from other cell types of the embryo, frozen, and freeze-dried on the TEM grids. The contents of electron-dense granules in the spiculogenic cells were shown by electron diffraction to be composed of amorphous calcium carbonate. These observations are consistent with the notion that the amorphous calcium carbonate-containing granules contain the precursor mineral phase for spicule formation and that the membrane surrounding the forming spicule is involved both in transport of material and in controlling spicule mineralization. Copyright 1999 Academic Press.

Nanoscale patterning of STIM1 and Orai1 during store-operated Ca2+ entry.

PubMed

Perni, Stefano; Dynes, Joseph L; Yeromin, Andriy V; Cahalan, Michael D; Franzini-Armstrong, Clara

2015-10-06

Stromal interacting molecule (STIM) and Orai proteins constitute the core machinery of store-operated calcium entry. We used transmission and freeze-fracture electron microscopy to visualize STIM1 and Orai1 at endoplasmic reticulum (ER)-plasma membrane (PM) junctions in HEK 293 cells. Compared with control cells, thin sections of STIM1-transfected cells possessed far more ER elements, which took the form of complex stackable cisternae and labyrinthine structures adjoining the PM at junctional couplings (JCs). JC formation required STIM1 expression but not store depletion, induced here by thapsigargin (TG). Extended molecules, indicative of STIM1, decorated the cytoplasmic surface of ER, bridged a 12-nm ER-PM gap, and showed clear rearrangement into small clusters following TG treatment. Freeze-fracture replicas of the PM of Orai1-transfected cells showed extensive domains packed with characteristic "particles"; TG treatment led to aggregation of these particles into sharply delimited "puncta" positioned upon raised membrane subdomains. The size and spacing of Orai1 channels were consistent with the Orai crystal structure, and stoichiometry was unchanged by store depletion, coexpression with STIM1, or an Orai1 mutation (L273D) affecting STIM1 association. Although the arrangement of Orai1 channels in puncta was substantially unstructured, a portion of channels were spaced at ∼15 nm. Monte Carlo analysis supported a nonrandom distribution for a portion of channels spaced at ∼15 nm. These images offer dramatic, direct views of STIM1 aggregation and Orai1 clustering in store-depleted cells and provide evidence for the interaction of a single Orai1 channel with small clusters of STIM1 molecules.

Polyhedral protein cages encase synaptic vesicles and participate in their attachment to the active zone.

PubMed

Zampighi, G A; Fisher, R S

1997-08-01

In an effort to elucidate the interactions between synaptic vesicles and the membrane of the active zone, we have investigated the structure of interneuronal asymmetric synapses in the neocortex of adult rats using thin-sectioning, freeze-fracture, and negative staining electron microscopy. We identified three subtypes of spherical synaptic vesicles. Type I were agranular vesicles of 47.5 +/- 3.8 nm (mean SD, n = 24) in diameter usually seen aggregated in clusters in the presynaptic bouton. Type II synaptic vesicles were composed of a approximately 45-nm-diameter lipid bilayer sphere encased in a cage 77 +/- 4.6 nm (mean SD, n = 42) in diameter. The cage was composed of open-faced pentamers 20-22 nm/side arranged as a regular polyhedron. Type II caged vesicles were found in clusters at the boutons, adhered to the active zone, and were also present in axons. Type III synaptic vesicles appeared as electron-dense spheres 60-75 nm in diameter abutted to the membrane of the active zone. Clathrin-coated vesicles and pits of 116.6 +/- 9 nm (mean SD, n = 14) in diameter were also present in both the pre- and postsynaptic sides. Freeze-fracture showed that some intrinsic membrane proteins in the active zone were arranged as pentamers exhibiting the same dimension of those forming cages (approximately 22 nm/side). From these data, we concluded that: (a) the presynaptic bouton contains a heterogeneous population of "caged" and "plain" synaptic vesicles and (b) type II synaptic vesicles bind to receptors in the active zone. Therefore, current models of transmitter release should take into account the substantial heterogeneity of the vesicle population and the binding of vesicular cages to the membrane of the active zone.

Study on the Effect of Straw Fiber on the Performance of Volcanic Slag Concrete

NASA Astrophysics Data System (ADS)

Xiao, Li-guang; Liu, Xi-xu

2018-03-01

In this paper, the effects of straw fiber on the working performance, mechanical properties and frost resistance of volcanic slag lightweight aggregate concrete were studied. The experimental results show that the straw fiber is subjected to surface carbonization treatment and mixed into the volcanic slag light aggregate concrete. The flexural strength and fracture pressure ratio of volcanic slag lightweight aggregate concrete are improved obviously Improved volcanic slag lightweight aggregate concrete brittleness improves toughness. Carbonized straw fiber greatly improves the frost resistance of volcanic slag lightweight aggregate concrete. So that the volcanic slag light aggregate concrete freeze-thaw cycle can reach 300 times.

Clastic polygonal networks around Lyot crater, Mars: Possible formation mechanisms from morphometric analysis

NASA Astrophysics Data System (ADS)

Brooker, L. M.; Balme, M. R.; Conway, S. J.; Hagermann, A.; Barrett, A. M.; Collins, G. S.; Soare, R. J.

2018-03-01

Polygonal networks of patterned ground are a common feature in cold-climate environments. They can form through the thermal contraction of ice-cemented sediment (i.e. formed from fractures), or the freezing and thawing of ground ice (i.e. formed by patterns of clasts, or ground deformation). The characteristics of these landforms provide information about environmental conditions. Analogous polygonal forms have been observed on Mars leading to inferences about environmental conditions. We have identified clastic polygonal features located around Lyot crater, Mars (50°N, 30°E). These polygons are unusually large (>100 m diameter) compared to terrestrial clastic polygons, and contain very large clasts, some of which are up to 15 metres in diameter. The polygons are distributed in a wide arc around the eastern side of Lyot crater, at a consistent distance from the crater rim. Using high-resolution imaging data, we digitised these features to extract morphological information. These data are compared to existing terrestrial and Martian polygon data to look for similarities and differences and to inform hypotheses concerning possible formation mechanisms. Our results show the clastic polygons do not have any morphometric features that indicate they are similar to terrestrial sorted, clastic polygons formed by freeze-thaw processes. They are too large, do not show the expected variation in form with slope, and have clasts that do not scale in size with polygon diameter. However, the clastic networks are similar in network morphology to thermal contraction cracks, and there is a potential direct Martian analogue in a sub-type of thermal contraction polygons located in Utopia Planitia. Based upon our observations, we reject the hypothesis that polygons located around Lyot formed as freeze-thaw polygons and instead an alternative mechanism is put forward: they result from the infilling of earlier thermal contraction cracks by wind-blown material, which then became compressed and/or cemented resulting in a resistant fill. Erosion then leads to preservation of these polygons in positive relief, while later weathering results in the fracturing of the fill material to form angular clasts. These results suggest that there was an extensive area of ice-rich terrain, the extent of which is linked to ejecta from Lyot crater.

Mechanical weathering and rock erosion by climate-dependent subcritical cracking

NASA Astrophysics Data System (ADS)

Eppes, Martha-Cary; Keanini, Russell

2017-06-01

This work constructs a fracture mechanics framework for conceptualizing mechanical rock breakdown and consequent regolith production and erosion on the surface of Earth and other terrestrial bodies. Here our analysis of fracture mechanics literature explicitly establishes for the first time that all mechanical weathering in most rock types likely progresses by climate-dependent subcritical cracking under virtually all Earth surface and near-surface environmental conditions. We substantiate and quantify this finding through development of physically based subcritical cracking and rock erosion models founded in well-vetted fracture mechanics and mechanical weathering, theory, and observation. The models show that subcritical cracking can culminate in significant rock fracture and erosion under commonly experienced environmental stress magnitudes that are significantly lower than rock critical strength. Our calculations also indicate that climate strongly influences subcritical cracking—and thus rock weathering rates—irrespective of the source of the stress (e.g., freezing, thermal cycling, and unloading). The climate dependence of subcritical cracking rates is due to the chemophysical processes acting to break bonds at crack tips experiencing these low stresses. We find that for any stress or combination of stresses lower than a rock's critical strength, linear increases in humidity lead to exponential acceleration of subcritical cracking and associated rock erosion. Our modeling also shows that these rates are sensitive to numerous other environment, rock, and mineral properties that are currently not well characterized. We propose that confining pressure from overlying soil or rock may serve to suppress subcritical cracking in near-surface environments. These results are applicable to all weathering processes.

The implications of tides on the Mimas ocean hypothesis

NASA Astrophysics Data System (ADS)

Rhoden, Alyssa Rose; Henning, Wade; Hurford, Terry A.; Patthoff, D. Alex; Tajeddine, Radwan

2017-02-01

We investigate whether a present-day global ocean within Mimas is compatible with the lack of tectonic activity on its surface by computing tidal stresses for ocean-bearing interior structure models derived from observed librations. We find that, for the suite of compatible rheological models, peak surface tidal stresses caused by Mimas' high eccentricity would range from a factor of 2 smaller to an order of magnitude larger than those on tidally active Europa. Thermal stresses from a freezing ocean, or a past higher eccentricity, would enhance present-day tidal stresses, exceeding the magnitudes associated with Europa's ubiquitous tidally driven fractures and, in some cases, the failure strength of ice in laboratory studies. Therefore, in order for Mimas to have an ocean, its ice shell cannot fail at the stress values implied for Europa. Furthermore, if Mimas' ocean is freezing out, the ice shell must also be able to withstand thermal stresses that could be an order of magnitude higher than the failure strength of laboratory ice samples. In light of these challenges, we consider an ocean-free Mimas to be the most straightforward model, best supported by our tidal stress analysis.

The Implications of Tides on the Mimas Ocean Hypothesis

NASA Technical Reports Server (NTRS)

Rhoden, Alyssa Rose; Henning, Wade; Hurford, Terry A.; Patthoff, D. Alex; Tajeddine, Radwan

2017-01-01

We investigate whether a present-day global ocean within Mimas is compatible with the lack of tectonic activity on its surface by computing tidal stresses for ocean-bearing interior structure models derived from observed librations. We find that, for the suite of compatible rheological models, peak surface tidal stresses caused by Mimas' high eccentricity would range from a factor of 2 smaller to an order of magnitude larger than those on tidally active Europa. Thermal stresses from a freezing ocean, or a past higher eccentricity, would enhance present-day tidal stresses, exceeding the magnitudes associated with Europa's ubiquitous tidally driven fractures and, in some cases, the failure strength of ice in laboratory studies. Therefore, in order for Mimas to have an ocean, its ice shell cannot fail at the stress values implied for Europa. Furthermore, if Mimas' ocean is freezing out, the ice shell must also be able to withstand thermal stresses that could be an order of magnitude higher than the failure strength of laboratory ice samples. In light of these challenges, we consider an ocean-free Mimas to be the most straightforward model, best supported by our tidal stress analysis.

Endoplasmic Reticulum-Golgi Intermediate Compartment Membranes and Vimentin Filaments Participate in Vaccinia Virus Assembly

PubMed Central

Risco, Cristina; Rodríguez, Juan R.; López-Iglesias, Carmen; Carrascosa, José L.; Esteban, Mariano; Rodríguez, Dolores

2002-01-01

Vaccinia virus (VV) has a complex morphogenetic pathway whose first steps are poorly characterized. We have studied the early phase of VV assembly, when viral factories and spherical immature viruses (IVs) form in the cytoplasm of the infected cell. After freeze-substitution numerous cellular elements are detected around assembling viruses: membranes, ribosomes, microtubules, filaments, and unidentified structures. A double membrane is clearly resolved in the VV envelope for the first time, and freeze fracture reveals groups of tubules interacting laterally on the surface of the viroplasm foci. These data strongly support the hypothesis of a cellular tubulovesicular compartment, related to the endoplasmic reticulum-Golgi intermediate compartment (ERGIC), as the origin of the first VV envelope. Moreover, the cytoskeletal vimentin intermediate filaments are found around viral factories and inside the viroplasm foci, where vimentin and the VV core protein p39 colocalize in the areas where crescents protrude. Confocal microscopy showed that ERGIC elements and vimentin filaments concentrate in the viral factories. We propose that modified cellular ERGIC membranes and vimentin intermediate filaments act coordinately in the construction of viral factories and the first VV form through a unique mechanism of viral morphogenesis from cellular elements. PMID:11799179

Electron tomography and cryo-SEM characterization reveals novel ultrastructural features of host-parasite interaction during Chlamydia abortus infection.

PubMed

Wilkat, M; Herdoiza, E; Forsbach-Birk, V; Walther, P; Essig, A

2014-08-01

Chlamydia (C.) abortus is a widely spread pathogen among ruminants that can be transmitted to women during pregnancy leading to severe systemic infection with consecutive abortion. As a member of the Chlamydiaceae, C. abortus shares the characteristic feature of an obligate intracellular biphasic developmental cycle with two morphological forms including elementary bodies (EBs) and reticulate bodies (RBs). In contrast to other chlamydial species, C. abortus ultrastructure has not been investigated yet. To do so, samples were fixed by high-pressure freezing and processed by different electron microscopic methods. Freeze-substituted samples were analysed by transmission electron microscopy, scanning transmission electron microscopical tomography and immuno-electron microscopy, and freeze-fractured samples were analysed by cryo-scanning electron microscopy. Here, we present three ultrastructural features of C. abortus that have not been reported up to now. Firstly, the morphological evidence that C. abortus is equipped with the type three secretion system. Secondly, the accumulation and even coating of whole inclusion bodies by membrane complexes consisting of multiple closely adjacent membranes which seems to be a C. abortus specific feature. Thirdly, the formation of small vesicles in the periplasmic space of RBs in the second half of the developmental cycle. Concerning the time point of their formation and the fact that they harbour chlamydial components, these vesicles might be morphological correlates of an intermediate step during the process of redifferentiation of RBs into EBs. As this feature has also been shown for C. trachomatis and C. pneumoniae, it might be a common characteristic of the family of Chlamydiaceae.

Determination of Stress Intensity Factor Distributions for "Interface" Cracks in Incompressible, Dissimilar Materials

NASA Technical Reports Server (NTRS)

Smith, C. W.

1997-01-01

The present study was undertaken in order to develop test methods and procedures for measuring the variation of the stress intensity factor through the thickness in bimaterial specimens containing cracks within and parallel to the bond line using the frozen stress photoelastic method. Since stress freezing materials are incompressible above critical temperature, and since thick plates are to be employed which tend to produce a state of plane strain near the crack tip, the interface near tip fracture equations reduce to the classic form for homogeneous materials. Moreover, zero thickness interfaces do not exist when materials are bonded together. It was decided early on that it would be important to insure a uniform straight and accurate crack tip region through the thickness of the body to reduce scatter in the SIF distribution through the thickness. It was also observed that rubberlike materials which were desired to be modeled exhibited significant tip blunting prior to crack extension and that some blunting of the tip would provide a more realistic model. It should be noted that, in normal stress freezing photoelastic work, it is considered good practice to avoid utilizing data near bond lines in photoelastic models due to the bond line stresses which inevitably develop when two parts are bonded together. Thus, the present study involves certain exploratory aspects in deviating from standard practice in stress freezing work. With the above ideas in mind, several different test methods were investigated and are described in the following sections and appendices. The geometry selected for the program was a thick, edge cracked specimen containing a bond line.

The structure and effect of suture zones in the Larsen C Ice Shelf, Antarctica

NASA Astrophysics Data System (ADS)

McGrath, Daniel; Steffen, Konrad; Holland, Paul R.; Scambos, Ted; Rajaram, Harihar; Abdalati, Waleed; Rignot, Eric

2014-03-01

Ice shelf fractures frequently terminate where they encounter suture zones, regions of material heterogeneity that form between meteoric inflows in ice shelves. This heterogeneity can consist of marine ice, meteoric ice with modified rheological properties, or the presence of fractures. Here, we use radar observations on the Larsen C Ice Shelf, Antarctica, to investigate (i) the termination of a 25 km long rift in the Churchill Peninsula suture zone, which was found to contain 60 m of accreted marine ice, and (ii) the along-flow evolution of a suture zone originating at Cole Peninsula. We determine a steady state field of basal melting/freezing rates and apply it to a flowline model to delineate the along-flow evolution of layers within the ice shelf. The thickening surface wedge of locally accumulated meteoric ice, which likely has limited lateral variation in its mechanical properties, accounts for 60% of the total ice thickness near the calving front. Thus, we infer that the lower 40% of the ice column and the material heterogeneities present there are responsible for resisting fracture propagation and thereby delaying tabular calving events, as demonstrated in the >40 year time series leading up to the 2004/2005 calving event for Larsen C. This likely represents a highly sensitive aspect of ice shelf stability, as changes in the oceanic forcing may lead to the loss of this heterogeneity.

Gap junctions are selectively associated with interlocking ball-and-sockets but not protrusions in the lens.

PubMed

Biswas, Sondip K; Lee, Jai Eun; Brako, Lawrence; Jiang, Jean X; Lo, Woo-Kuen

2010-11-09

Ball-and-sockets and protrusions are specialized interlocking membrane domains between lens fibers of all species studied. Ball-and-sockets and protrusions are similar in their shape, size, and surface morphology, and are traditionally believed to play a key role in maintaining fiber-to-fiber stability. Here, we evaluate the hypothesis that ball-and-sockets and protrusions possess important structural and functional differences during fiber cell differentiation and maturation. Intact lenses of leghorn chickens (E7 days to P62 weeks old) and rhesus monkeys (1.5-20 years old) were studied with SEM, freeze-fracture TEM, freeze-fracture immunogold labeling (FRIL), and filipin cytochemistry for membrane cholesterol detection. SEM showed that ball-and-sockets were distributed along the long and short sides of hexagonal fiber cells, whereas protrusions were located along the cell corners, from superficial to deep cortical regions in both chicken and monkey lenses. Importantly, by freeze-fracture TEM, we discovered the selective association of gap junctions with all ball-and-sockets examined, but not with protrusions, in both species. In the embryonic chicken lens (E18), the abundant distribution of ball-and-socket gap junctions was regularly found in an approximate zone extending at least 300 μm deep from the equatorial surface of the superficial cortical fibers. Many ball-and-socket gap junctions often protruded deeply into neighboring cells. However, in the mature fibers of monkey lenses, several ball-and-sockets exhibited only partial occupancy of gap junctions with disorganized connexons, possibly due to degradation of gap junctions during fiber maturation and aging. FRIL analysis confirmed that both connexin46 (Cx46) and connexin50 (Cx50) antibodies specifically labeled ball-and-socket gap junctions, but not protrusions. Furthermore, filipin cytochemistry revealed that the ball-and-socket gap junctions contained different amounts of cholesterol (i.e., cholesterol-rich versus cholesterol-free) as seen with the filipin-cholesterol-complexes (FCC) in different cortical regions during maturation. In contrast, the protrusions contained consistently high cholesterol amounts (i.e., 402 FCCs/μm2 membrane) which were approximately two times greater than that of the cholesterol-rich gap junctions (i.e., 188 FCCs/μm2 membrane) found in ball-and-sockets. Gap junctions are regularly associated with all ball-and-sockets examined in metabolically active young cortical fibers, but not with protrusions, in both chicken and monkey lenses. Since these unique gap junctions often protrude deeply into neighboring cells to increase membrane surface areas, they may significantly facilitate cell-to-cell communication between young cortical fiber cells. In particular, the large number of ball-and-socket gap junctions found near the equatorial region may effectively facilitate the flow of outward current toward the equatorial surface for internal circulation of ions in the lens. In contrast, a consistent distribution of high concentrations of cholesterol in protrusions would make the protrusion membrane less deformable and would be more suitable for maintaining fiber-to-fiber stability during visual accommodation. Thus, the ball-and-sockets and protrusions are two structurally and functionally distinct membrane domains in the lens.

Gap junctions are selectively associated with interlocking ball-and-sockets but not protrusions in the lens

PubMed Central

Biswas, Sondip K.; Lee, Jai Eun; Brako, Lawrence; Jiang, Jean X.

2010-01-01

Purpose Ball-and-sockets and protrusions are specialized interlocking membrane domains between lens fibers of all species studied. Ball-and-sockets and protrusions are similar in their shape, size, and surface morphology, and are traditionally believed to play a key role in maintaining fiber-to-fiber stability. Here, we evaluate the hypothesis that ball-and-sockets and protrusions possess important structural and functional differences during fiber cell differentiation and maturation. Methods Intact lenses of leghorn chickens (E7 days to P62 weeks old) and rhesus monkeys (1.5–20 years old) were studied with SEM, freeze-fracture TEM, freeze-fracture immunogold labeling (FRIL), and filipin cytochemistry for membrane cholesterol detection. Results SEM showed that ball-and-sockets were distributed along the long and short sides of hexagonal fiber cells, whereas protrusions were located along the cell corners, from superficial to deep cortical regions in both chicken and monkey lenses. Importantly, by freeze-fracture TEM, we discovered the selective association of gap junctions with all ball-and-sockets examined, but not with protrusions, in both species. In the embryonic chicken lens (E18), the abundant distribution of ball-and-socket gap junctions was regularly found in an approximate zone extending at least 300 μm deep from the equatorial surface of the superficial cortical fibers. Many ball-and-socket gap junctions often protruded deeply into neighboring cells. However, in the mature fibers of monkey lenses, several ball-and-sockets exhibited only partial occupancy of gap junctions with disorganized connexons, possibly due to degradation of gap junctions during fiber maturation and aging. FRIL analysis confirmed that both connexin46 (Cx46) and connexin50 (Cx50) antibodies specifically labeled ball-and-socket gap junctions, but not protrusions. Furthermore, filipin cytochemistry revealed that the ball-and-socket gap junctions contained different amounts of cholesterol (i.e., cholesterol-rich versus cholesterol-free) as seen with the filipin-cholesterol-complexes (FCC) in different cortical regions during maturation. In contrast, the protrusions contained consistently high cholesterol amounts (i.e., 402 FCCs/μm2 membrane) which were approximately two times greater than that of the cholesterol-rich gap junctions (i.e., 188 FCCs/μm2 membrane) found in ball-and-sockets. Conclusions Gap junctions are regularly associated with all ball-and-sockets examined in metabolically active young cortical fibers, but not with protrusions, in both chicken and monkey lenses. Since these unique gap junctions often protrude deeply into neighboring cells to increase membrane surface areas, they may significantly facilitate cell-to-cell communication between young cortical fiber cells. In particular, the large number of ball-and-socket gap junctions found near the equatorial region may effectively facilitate the flow of outward current toward the equatorial surface for internal circulation of ions in the lens. In contrast, a consistent distribution of high concentrations of cholesterol in protrusions would make the protrusion membrane less deformable and would be more suitable for maintaining fiber-to-fiber stability during visual accommodation. Thus, the ball-and-sockets and protrusions are two structurally and functionally distinct membrane domains in the lens. PMID:21139982

Intracellular ice and cell survival in cryo-exposed embryonic axes of recalcitrant seeds of Acer saccharinum: an ultrastructural study of factors affecting cell and ice structures

PubMed Central

Wesley-Smith, James; Berjak, Patricia; Pammenter, N. W.; Walters, Christina

2014-01-01

Background and Aims Cryopreservation is the only long-term conservation strategy available for germplasm of recalcitrant-seeded species. Efforts to cryopreserve this form of germplasm are hampered by potentially lethal intracellular freezing events; thus, it is important to understand the relationships among cryo-exposure techniques, water content, structure and survival. Methods Undried embryonic axes of Acer saccharinum and those rapidly dried to two different water contents were cooled at three rates and re-warmed at two rates. Ultrastructural observations were carried out on radicle and shoot tips prepared by freeze-fracture and freeze-substitution to assess immediate (i.e. pre-thaw) responses to cooling treatments. Survival of axes was assessed in vitro. Key Results Intracellular ice formation was not necessarily lethal. Embryo cells survived when crystal diameter was between 0·2 and 0·4 µm and fewer than 20 crystals were distributed per μm2 in the cytoplasm. Ice was not uniformly distributed within the cells. In fully hydrated axes cooled at an intermediate rate, the interiors of many organelles were apparently ice-free; this may have prevented the disruption of vital intracellular machinery. Intracytoplasmic ice formation did not apparently impact the integrity of the plasmalemma. The maximum number of ice crystals was far greater in shoot apices, which were more sensitive than radicles to cryo-exposure. Conclusions The findings challenge the accepted paradigm that intracellular ice formation is always lethal, as the results show that cells can survive intracellular ice if crystals are small and localized in the cytoplasm. Further understanding of the interactions among water content, cooling rate, cell structure and ice structure is required to optimize cryopreservation treatments without undue reliance on empirical approaches. PMID:24368198

Intracellular ice and cell survival in cryo-exposed embryonic axes of recalcitrant seeds of Acer saccharinum: an ultrastructural study of factors affecting cell and ice structures.

PubMed

Wesley-Smith, James; Berjak, Patricia; Pammenter, N W; Walters, Christina

2014-03-01

Cryopreservation is the only long-term conservation strategy available for germplasm of recalcitrant-seeded species. Efforts to cryopreserve this form of germplasm are hampered by potentially lethal intracellular freezing events; thus, it is important to understand the relationships among cryo-exposure techniques, water content, structure and survival. Undried embryonic axes of Acer saccharinum and those rapidly dried to two different water contents were cooled at three rates and re-warmed at two rates. Ultrastructural observations were carried out on radicle and shoot tips prepared by freeze-fracture and freeze-substitution to assess immediate (i.e. pre-thaw) responses to cooling treatments. Survival of axes was assessed in vitro. Intracellular ice formation was not necessarily lethal. Embryo cells survived when crystal diameter was between 0·2 and 0·4 µm and fewer than 20 crystals were distributed per μm(2) in the cytoplasm. Ice was not uniformly distributed within the cells. In fully hydrated axes cooled at an intermediate rate, the interiors of many organelles were apparently ice-free; this may have prevented the disruption of vital intracellular machinery. Intracytoplasmic ice formation did not apparently impact the integrity of the plasmalemma. The maximum number of ice crystals was far greater in shoot apices, which were more sensitive than radicles to cryo-exposure. The findings challenge the accepted paradigm that intracellular ice formation is always lethal, as the results show that cells can survive intracellular ice if crystals are small and localized in the cytoplasm. Further understanding of the interactions among water content, cooling rate, cell structure and ice structure is required to optimize cryopreservation treatments without undue reliance on empirical approaches.

Resting myosin cross-bridge configuration in frog muscle thick filaments.

PubMed

Cantino, M; Squire, J

1986-02-01

Clear images of myosin filaments have been seen in shadowed freeze-fracture replicas of single fibers of relaxed frog semitendinosus muscles rapidly frozen using a dual propane jet freezing device. These images have been analyzed by optical diffraction and computer averaging and have been modelled to reveal details of the myosin head configuration on the right-handed, three-stranded helix of cross-bridges. Both the characteristic 430-A and 140-150-A repeats of the myosin cross-bridge array could be seen. The measured filament backbone diameter was 140-160 A, and the outer diameter of the cross-bridge array was 300 A. Evidence is presented that suggests that the observed images are consistent with a model in which both of the heads of one myosin molecule tilt in the same direction at an angle of approximately 50-70 degrees to the normal to the filament long axis and are slewed so that they lie alongside each other and their radially projected density lies along the three right-handed helical tracks. Any perturbation of the myosin heads away from their ideal lattice sites needed to account for x-ray reflections not predicted for a perfect helix must be essentially along the three helical tracks of cross-bridges. Little trace of the presence of non-myosin proteins could be seen.

Isolation, Chemical Composition, and Ultrastructural Features of the Cell Membrane of the Mycoplasma-Like Organism Spiroplasma citri

PubMed Central

Razin, Shmuel; Hasin, Miriam; Ne'eman, Zvi; Rottem, Shlomo

1973-01-01

Thin sections of Spiroplasma citri, a mycoplasma-like organism isolated from citrus infected with “Stubborn” disease, showed the organisms to be limited by a single trilaminar plasma membrane. An additional outer layer could, however, be frequently seen in freeze-etched preparations of unwashed cells. The organisms were found to be extremely sensitive to lysis by osmotic shock. The cell membrane of S. citri isolated in this way resembled that of mycoplasmas in ultrastructure and gross chemical composition. The isolated membranes showed the characteristic trilaminar shape in section and the typical particle-studded fracture faces in freeze-etched preparations. Protein and lipid formed over 80% of the total dry weight of the membrane, which had a density of ~1.180 g/cm3. Cholesterol constituted over 20% of the total membrane lipid. Phosphatidyl-glycerol, synthesized by the organisms, was the major phospholipid. Significant amounts of hexosamine (15 to 35 μg/mg of membrane protein) could be found in the membrane preparations. Our results support the thesis that S. citri does not possess a cell wall, either of the gram-positive or the gram-negative type, though it may be coated by some other type of an envelope or by a slime layer, at least temporarily. Images PMID:4127633

High-linearity piezoresistive response of mechanically strong graphene-based elastomer

NASA Astrophysics Data System (ADS)

Yuanzheng, Luo; Buyin, Li; Xiaoqi

2017-05-01

Traditional additive-free graphene bulk materials based on mono- three dimensional(3D) graphene networks type are fragile in most cases, which is unfavorable for their potential applications. Here we present compressible graphene foams (CGF) with superior properties endowed by the hierarchical porous structure, which taking graphene sheets as an inorganic embedding material and polyurethane sponge (PUS) as a polymer open-framework. The preparation process utilized a dip-coating method associated with directional freezing followed by lyophilization. The as-synthesized CGF not only possess a combination of ultralow density and excellent electrical conductivity, but it also can withstand large strains (>99%) without permanent deformation or fracture. We believe that these sponge/graphene embeddable multifunctional nanocomposites will expand practical applications of graphene monolith in the future.

Chiral lyotropic chromonic liquid crystals composed of disodium cromoglycate doped with water-soluble chiral additives.

PubMed

Shirai, Tatsuya; Shuai, Min; Nakamura, Keita; Yamaguchi, Akihiro; Naka, Yumiko; Sasaki, Takeo; Clark, Noel A; Le, Khoa V

2018-02-28

We investigated the pitches of cholesteric liquid crystals prepared by mixing disodium cromoglycate (DSCG) in water with 5 different water-soluble chiral additives. The measurements are based on the Grandjean-Cano wedge cell method. Overall, the twisting effect is weak, and the shortest pitch of 2.9 ± 0.2 μm is obtained using trans-4-hydroxy-l-proline, by which the cholesteric sample is iridescent at certain viewing angles. Freeze-fracture transmission electron microscopy (FFTEM) was also performed for the first time on both the nematic and cholesteric phases, revealing that stacked chromonic aggregates are very long, up to a few hundred nm, which explains why cholesteric chromonic liquid crystals hardly have pitches in the visible wavelength region.

Process for controlling morphology and improving thermal-mechanical performance of high performance interpenetrating and semi-interpenetrating polymer networks

NASA Technical Reports Server (NTRS)

Pater, Ruth H. (Inventor); Hansen, Marion G. (Inventor)

1997-01-01

In the process of the present invention, a non-polar, aprotic solvent is removed from an oligomer/polymer solution by freeze-drying in order to produce IPNs and semi-IPNs. By thermally quenching the solution to a solid in a short length of time, the size of the minor constituent-rich regions is greatly reduced as they are excluded along with the major constituent from the regions of crystallizing solvent. The use of this process sequence of controlling phase morphology provides IPNs and semi-IPNs with improved fracture toughness, microcracking resistance, and other physical-mechanical properties as compared to IPNs and semi-IPNs formed when the solvent is evaporated rather than sublimed.

Process for controlling morphology and improving thermal mechanical performance of high performance interpenetrating and semiinterpenetrating polymer networks

NASA Technical Reports Server (NTRS)

Pater, Ruth H. (Inventor); Hansen, Marion G. (Inventor)

1998-01-01

In the process of the present invention, a non-polar, aprotic solvent is removed from an oligomer/polymer solution by freeze-drying in order to produce IPNs and semi-IPNs. By thermally quenching the solution to a solid in a short length of time, the size of the minor constituent-rich regions is greatly reduced as they are excluded along with the major constituent from the regions of crystallizing solvent. The use of this process sequence of controlling phase morphology provides IPNs and semi-IPNs with improved fracture toughness, microcracking resistance, and other physical-mechanical properties as compared to IPNs and semi-IPNs formed when the solvent is evaporated rather than sublimed.

Annular Air Leaks in a liquid hydrogen storage tank

NASA Astrophysics Data System (ADS)

Krenn, AG; Youngquist, RC; Starr, SO

2017-12-01

Large liquid hydrogen (LH2) storage tanks are vital infrastructure for NASA, the DOD, and industrial users. Over time, air may leak into the evacuated, perlite filled annular region of these tanks. Once inside, the extremely low temperatures will cause most of the air to freeze. If a significant mass of air is allowed to accumulate, severe damage can result from nominal draining operations. Collection of liquid air on the outer shell may chill it below its ductility range, resulting in fracture. Testing and analysis to quantify the thermal conductivity of perlite that has nitrogen frozen into its interstitial spaces and to determine the void fraction of frozen nitrogen within a perlite/frozen nitrogen mixture is presented. General equations to evaluate methods for removing frozen air, while avoiding fracture, are developed. A hypothetical leak is imposed on an existing tank geometry and a full analysis of that leak is detailed. This analysis includes a thermal model of the tank and a time-to-failure calculation. Approaches to safely remove the frozen air are analyzed, leading to the conclusion that the most feasible approach is to allow the frozen air to melt and to use a water stream to prevent the outer shell from chilling.

Ribbed moraine stratigraphy and formation in southern Finnish Lapland

NASA Astrophysics Data System (ADS)

Sarala, Pertti

2006-05-01

Characteristics of ribbed moraines, the dominating moraine type in southern Finnish Lapland, have been studied in detail. The ridges are composed of several till units, of which the bottommost units consist of mature basal tills and the surficial parts are enriched with local, short-transport rock fragments and boulders in till and at the surface of ridges. As a result of this re-examination a two-step model of the formation process of ribbed moraines is presented. In the first stage, while cold-based conditions prevailed, both the bottommost part of the ice sheet and the frozen, substrate fractured under compressive ice flow. Following glacial transport of fractured blocks and formation of the transverse ridge morphology, erosion between the ridges continued owing to freeze-thaw process under variable pressure conditions. In the areas with a low pre-existing till sheet, the process caused quarrying of the bedrock surface and subsequent deposition of rock fragments and boulders under high pressure on the next ridge. The most suitable conditions for ribbed moraine formation existed during Late Weichselian deglaciation, after the Younger Dryas when the climate warmed very quickly, leading to an imbalance between a warm glacier surface and a cold base. Copyright

Membrane interactions between secretion granules and plasmalemma in three exocrine glands

PubMed Central

Tanaka, Y; De Camilli, P; Meldolesi, J

1980-01-01

Three types of membrane interactions were studied in three exocrine systems (the acinar cells of the rat parotid, rat lacrimal gland, and guinea pig pancrease) by freeze- fracture and thin-section electron microscopy: exocytosis, induced in vivo by specific pharmacological stimulations; the mutual apposition of secretory granule membranes in the intact cell; membrane appositions induced in vitro by centrifugation of the isolated granules. In all three glandular cells, the distribution of intramembrane particles (IMP) on the fracture faces of the luminal plasmagranule membrane particles (IMP) on the fracture faces of the lumenal plasmalemma appeared random before stimulation. However, after injection of secretagogues, IMP were rapidly clearly from the areas of granule- plasmalemma apposition in the parotid cells and, especially, in lacrimocytes. In the latter, the cleared areas appeared as large bulges toward the lumen, whereas in the parotid they were less pronounced. Exocytotic openings were usually large and the fracture faces of their rims were covered with IMP. In contrast, in stimulated pancreatic acinar cells, the IMP distribution remained apparently random after stimulation. Exocytoses were established through the formation of narrown necks, and no images which might correspond to early stages of membrane fusion were revealed. Within the cytoplasm of parotid and lacrimal cells (but not in the pancreas), both at rest and after stimulation, secretion granules were often closely apposed by means of flat, circular areas, also devoid of IMP. In thin sections, the images corresponding to IMP-free areas were close granule-granule and granule-plasmalemma appositions, sometimes with focal merging of the membrane outer layers to yield pentalaminar structures. Isolated secretion granules were forced together in vitro by centrifugation. Under these conditions, increasing the centrifugal force from 1,600 to 50,000 g for 10 min resulted in a progressive, statistically significant increase of the frequency of IMP-free flat appositions between parotid granules. In contrast, no such areas were seen between freeze-fractured pancreatic granules, although some focal pentalaminar appositions appeared in section after centrifugation at 50 and 100,000 g for 10 min. On the basis of the observation that, in secretory cells, IMP clearing always develops in deformed membrane areas (bulges, depressions, flat areas), it is suggested that it might result from the forced mechanical apposition of the interacting membranes. This might be a preliminary process not sufficient to initiate fusion. In the pancreas, IMP clearing could occur over surface areas too small to be detected. In stimulated parotid and lacrimal glands they were exceptional. These structures were either attached at the sites of continuity between granule and plasma membranes, or free in the acinar lumen, with a preferential location within exocytotic pockets or in their proximity. Experiments designed to investigate the nature of these blisters and vesicles revealed that they probably arise artifactually during glutaraldehyde fixation. In fact, (a) they were large and numerous in poorly fixed samples but were never observed in thin sections of specimens fixed in one step with glutaraldehyde and OsO(4); and (b) no increase in concentration of phospholipids was observed in the parotid saliva and pancreatic juice after stimulation of protein discharge, as was to be expected if release of membrane material were occurring after exocytosis. PMID:7380885

[Ultrastructural organization of cytoplasmatic membrane of Anaerobacter polyendosporus studied by electron microscopic cryofractography].

PubMed

Duda, V I; Suzina, N E; Dmitriev, V V

2001-01-01

Anaerobacter polyendosporus cells do not have typical mesosomes. However, the analysis of this anaerobic multispore bacterium by electron microscopic cryofractography showed that its cytoplasmic membrane contains specific intramembrane structures in the form of flat lamellar inverted lipid membranes tenths of nanometers to several microns in size. It was found that these structures are located in the hydrophobic interior between the outer and inner leaflets of the cytoplasmic membrane and do not contain intramembrane particles that are commonly present on freeze-fracture replicas. The flat inverted lipid membranes were revealed in bacterial cells cultivated under normal growth conditions, indicating the existence of a complex-type compartmentalization in biological membranes, which manifests itself in the formation of intramembrane compartments having the appearance of vesicles and inverted lipid membranes.

Triblock copolyampholytes from 5-(N,N-dimethyl amino)isoprene styrene, and methacrylic acid: Synthesis and solution properties

NASA Astrophysics Data System (ADS)

Bieringer, R.; Abetz, V.; Müller, A. H. E.

ABC triblock copolymers of the type poly[5-(N,N-dimethyl amino)isoprene]-block-polystyrene-block-poly(tert-butyl methacrylate) (AiST) were synthesized and hydrolyzed to yield poly[5-(N,N-dimethyl amino)isoprene]-block-polystyrene-block-poly(methacrylic acid) (AiSA) triblock copolyampholytes. Due to a complex solubility behavior the solution properties of these materials had to be investigated in THF/water solvent mixtures. Potentiometric titrations of AiSA triblock copolyampholytes showed two inflection points with the A block being deprotonated prior to the Ai hydrochloride block thus forming a polyzwitterion at the isoelectric point (iep). The aggregation behavior was studied by dynamic light scattering (DLS) and freeze-fracture/transmission electron microscopy (TEM). Large vesicular structures with almost pH-independent radii were observed.

Adrenocortical Gap Junctions and Their Functions

PubMed Central

Bell, Cheryl L.; Murray, Sandra A.

2016-01-01

Adrenal cortical steroidogenesis and proliferation are thought to be modulated by gap junction-mediated direct cell–cell communication of regulatory molecules between cells. Such communication is regulated by the number of gap junction channels between contacting cells, the rate at which information flows between these channels, and the rate of channel turnover. Knowledge of the factors regulating gap junction-mediated communication and the turnover process are critical to an understanding of adrenal cortical cell functions, including development, hormonal response to adrenocorticotropin, and neoplastic dedifferentiation. Here, we review what is known about gap junctions in the adrenal gland, with particular attention to their role in adrenocortical cell steroidogenesis and proliferation. Information and insight gained from electrophysiological, molecular biological, and imaging (immunocytochemical, freeze fracture, transmission electron microscopic, and live cell) techniques will be provided. PMID:27445985

Nanosized cancer cell-targeted polymeric immunomicelles loaded with superparamagnetic iron oxide nanoparticles

NASA Astrophysics Data System (ADS)

Sawant, Rishikesh M.; Sawant, Rupa R.; Gultepe, Evin; Nagesha, Dattatri; Papahadjopoulos-Sternberg, Brigitte; Sridhar, Srinivas; Torchilin, Vladimir P.

2009-10-01

Stable 30-50 nm polymeric polyethylene glycol-phosphatidylethanolamine (PEG-PE)-based micelles entrapping superparamagnetic iron oxide nanoparticles (SPION) have been prepared. At similar concentrations of SPION, the SPION-micelles had significantly better magnetic resonance imaging (MRI) T2 relaxation signal compared to `plain' SPION. Freeze-fracture electron microscopy confirmed SPION entrapment in the lipid core of the PEG-PE micelles. To enhance the targeting capability of these micelles, their surface was modified with the cancer cell-specific anti-nucleosome monoclonal antibody 2C5 (mAb 2C5). Such mAb 2C5-SPION immunomicelles demonstrated specific binding with cancer cells in vitro and were able to bring more SPION to the cancer cells thus demonstrating the potential to be used as targeted MRI contrast agents for tumor imaging.

A forensic investigation into the etiology of bat mortality at a wind farm: barotrauma or traumatic injury?

PubMed

Rollins, K E; Meyerholz, D K; Johnson, G D; Capparella, A P; Loew, S S

2012-03-01

Migrating bats have increased mortality near moving turbine blades at wind farms. The authors evaluated competing hypotheses of barotrauma and traumatic injury to determine the cause. They first examined the utility of lungs from salvaged bat carcasses for histopathologic diagnosis of barotrauma and studied laboratory mice as a model system. Postmortem time, environmental temperature, and freezing of carcasses all affected the development of vascular congestion, hemorrhage, and edema. These common tissue artifacts mimicked the diagnostic criteria of pulmonary barotrauma; therefore, lung tissues from salvaged bats should not be used for barotrauma diagnosis. The authors next compared wind farm (WF) bats to building collision (BC) bats collected near downtown Chicago buildings. WF bats had an increased incidence in fracture cases and specific bone fractures and had more external lacerations than BC bats. WF bats had additional features of traumatic injury, including diaphragmatic hernia, subcutaneous hemorrhage, and bone marrow emboli. In summary, 73% (190 of 262) of WF bats had lesions consistent with traumatic injury. The authors then examined for ruptured tympana, a sensitive marker of barotrauma in humans. BC bats had only 1 case (2%, 1 of 42), but this was attributed to concurrent cranial fractures, whereas WF bats had a 20% (16 of 81) incidence. When cases with concurrent traumatic injury were excluded, this yielded a small fraction (6%, 5 of 81) of WF bats with lesions possibly consistent with barotrauma etiology. Forensic pathology examination of the data strongly suggests that traumatic injury is the major cause of bat mortality at wind farms and, at best, barotrauma is a minor etiology.

Tidal reorientation and the fracturing of Jupiter's moon Europa

USGS Publications Warehouse

McEwen, A.S.

1986-01-01

The most striking characteristic of Europa is the network of long linear albedo markings over the surface, suggestive of global-scale tectonic processes. Various explanations for the fractures have been proposed: Freezing and expansion of an early liquid water ocean1, planetary expansion due to dehydration of hydrated silicates2, localization by weak points in the crust generated by impacts3, and a combination of stresses due to planetary volume change and tidal distortions from orbital recession and orbital eccentricity4,5. Calculations by Yoder6 and Greenberg and Weidenschilling7 have shown that Europa may rotate slightly more rapidly than the synchronous rate, with a rotation period (reorientation through 360??) ranging from 20 to >103 yr if a liquid mantle is present, or up to 1010 yr if the satellite is essentially solid7. Helfen-stein and Parmentier8 modelled the stresses due to nonsynchronous rotation, and concluded that this could explain the long fractures in part of the anti-jovian hemisphere. In this note, I present a global map of lineaments with long arc lengths (>20?? or 550 km), and compare the lineament orientations to the tensile stress trajectories due to tidal distortions (changes in the lengths of three principal semiaxes) and to nonsynchronous rotation (longitudinal reorientation of two of the principal semiaxes). An excellent orthogonal fit to the lineaments is achieved by the stresses due to nonsynchronous rotation with the axis radial to Jupiter located 25?? east of its present position. This fit suggests that nonsynchronous rotation occurred at some time in Europa's history. ?? 1986 Nature Publishing Group.

The effect of hydroxyapatite addition on the mechanical properties of polyvinyl alcohol/chitosan biomaterials for bone scaffolds application

NASA Astrophysics Data System (ADS)

Ramahdita, Ghiska; Puspita, Debie Maya; Albab, Muh Fadhil; Alfata, Rowi; Sofyan, Nofrijon; Yuwono, Akhmad Herman

2018-02-01

The increasing number of bone fracture incident in Indonesia from year to year needs an urgent problem solving of the limited bone substitute which can meet the necessary criteria for that purpose. Motivated by this, therefore, the current study is focusing on the optimization of material properties used as bone scaffold. A biomaterial of polyvinyl alcohol (PVA)/chitosan-hydroxyapatite (HA) composite was successfully made by wet chemistry method, followed by freeze thawing and freeze drying. For comparison purposes, the percentage of HA has been varied from 0, 25 and 40 % (wt/v). The resulting composites were characterized by Fourier transform infrared (FTIR) spectroscopy, scanning electron microscope (SEM), compressive test, and swelling behavior. The results showed that the addition of HA up to 40% (wt/v) has yielded a porous structure with an average pore size of 42.39 µm. In addition, the compressive modulus was enhanced from 14 MPa for 0% HA to 143, and 191 MPa for composites with the addition of HA from 25 to 40% (wt/v). The addition of HA has also reduced the swelling ratio from 296% for the sample without HA to 85 and 78 % for sample with addition of HA from 25 to 40 (wt/v), respectively. The obtained results show that PVA/chitosan-HA in this study is potential to be used as scaffold in bone tissue engineering.

Structure of the cellulose synthase complex of Gluconacetobacter hansenii at 23.4 Å resolution

DOE PAGES

Du, Juan; Vepachedu, Venkata; Cho, Sung Hyun; ...

2016-05-23

Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA) exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC) that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalytic AcsAB complex is embedded in the cytoplasmic membrane. The C-terminal portion of AcsC is predicted to form a translocation channel in the outer membrane, with the rest of AcsC possibly interacting with AcsDmore » in the periplasm. It is thus believed that synthesis from an organized array of TCs coordinated with extrusion by AcsC and AcsD enable this bacterium to make crystalline cellulose. The only structural data that exist for this system are the above mentioned freeze-fracture TEM images, fluorescence microscopy images revealing that TCs align in a row, a crystal structure of AcsD bound to cellopentaose, and a crystal structure of PilZ domain of AcsA. Here we advance our understanding of the structural basis for crystalline cellulose production by bacterial cellulose synthase by determining a negative stain structure resolved to 23.4 angstrom for highly purified AcsAB complex that catalyzed incorporation of UDP-glucose into β-1,4-glucan chains, and responded to the presence of allosteric activator cyclic diguanylate. Although the AcsAB complex was functional in vitro, the synthesized cellulose was not visible in TEM. The negative stain structure revealed that AcsAB is very similar to that of the BcsAB synthase of Rhodobacter sphaeroides, a non-crystalline cellulose producing bacterium. Furthermore, the results indicate that the crystalline cellulose producing and non-crystalline cellulose producing bacteria share conserved catalytic and membrane translocation components, and support the hypothesis that it is the extrusion mechanism and order in linearly arrayed TCs that enables production of crystalline cellulose.« less

Structure of the cellulose synthase complex of Gluconacetobacter hansenii at 23.4 Å resolution

DOE Office of Scientific and Technical Information (OSTI.GOV)

Du, Juan; Vepachedu, Venkata; Cho, Sung Hyun

Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA) exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC) that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalytic AcsAB complex is embedded in the cytoplasmic membrane. The C-terminal portion of AcsC is predicted to form a translocation channel in the outer membrane, with the rest of AcsC possibly interacting with AcsDmore » in the periplasm. It is thus believed that synthesis from an organized array of TCs coordinated with extrusion by AcsC and AcsD enable this bacterium to make crystalline cellulose. The only structural data that exist for this system are the above mentioned freeze-fracture TEM images, fluorescence microscopy images revealing that TCs align in a row, a crystal structure of AcsD bound to cellopentaose, and a crystal structure of PilZ domain of AcsA. Here we advance our understanding of the structural basis for crystalline cellulose production by bacterial cellulose synthase by determining a negative stain structure resolved to 23.4 angstrom for highly purified AcsAB complex that catalyzed incorporation of UDP-glucose into β-1,4-glucan chains, and responded to the presence of allosteric activator cyclic diguanylate. Although the AcsAB complex was functional in vitro, the synthesized cellulose was not visible in TEM. The negative stain structure revealed that AcsAB is very similar to that of the BcsAB synthase of Rhodobacter sphaeroides, a non-crystalline cellulose producing bacterium. Furthermore, the results indicate that the crystalline cellulose producing and non-crystalline cellulose producing bacteria share conserved catalytic and membrane translocation components, and support the hypothesis that it is the extrusion mechanism and order in linearly arrayed TCs that enables production of crystalline cellulose.« less

Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 Å Resolution

PubMed Central

Du, Juan; Vepachedu, Venkata; Cho, Sung Hyun; Kumar, Manish; Nixon, B. Tracy

2016-01-01

Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA) exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC) that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalytic AcsAB complex is embedded in the cytoplasmic membrane. The C-terminal portion of AcsC is predicted to form a translocation channel in the outer membrane, with the rest of AcsC possibly interacting with AcsD in the periplasm. It is thus believed that synthesis from an organized array of TCs coordinated with extrusion by AcsC and AcsD enable this bacterium to make crystalline cellulose. The only structural data that exist for this system are the above mentioned freeze-fracture TEM images, fluorescence microscopy images revealing that TCs align in a row, a crystal structure of AcsD bound to cellopentaose, and a crystal structure of PilZ domain of AcsA. Here we advance our understanding of the structural basis for crystalline cellulose production by bacterial cellulose synthase by determining a negative stain structure resolved to 23.4 Å for highly purified AcsAB complex that catalyzed incorporation of UDP-glucose into β-1,4-glucan chains, and responded to the presence of allosteric activator cyclic diguanylate. Although the AcsAB complex was functional in vitro, the synthesized cellulose was not visible in TEM. The negative stain structure revealed that AcsAB is very similar to that of the BcsAB synthase of Rhodobacter sphaeroides, a non-crystalline cellulose producing bacterium. The results indicate that the crystalline cellulose producing and non-crystalline cellulose producing bacteria share conserved catalytic and membrane translocation components, and support the hypothesis that it is the extrusion mechanism and order in linearly arrayed TCs that enables production of crystalline cellulose. PMID:27214134

Analytical solution and numerical simulation of the liquid nitrogen freezing-temperature field of a single pipe

NASA Astrophysics Data System (ADS)

Cai, Haibing; Xu, Liuxun; Yang, Yugui; Li, Longqi

2018-05-01

Artificial liquid nitrogen freezing technology is widely used in urban underground engineering due to its technical advantages, such as simple freezing system, high freezing speed, low freezing temperature, high strength of frozen soil, and absence of pollution. However, technical difficulties such as undefined range of liquid nitrogen freezing and thickness of frozen wall gradually emerge during the application process. Thus, the analytical solution of the freezing-temperature field of a single pipe is established considering the freezing temperature of soil and the constant temperature of freezing pipe wall. This solution is then applied in a liquid nitrogen freezing project. Calculation results show that the radius of freezing front of liquid nitrogen is proportional to the square root of freezing time. The radius of the freezing front also decreases with decreased the freezing temperature, and the temperature gradient of soil decreases with increased distance from the freezing pipe. The radius of cooling zone in the unfrozen area is approximately four times the radius of the freezing front. Meanwhile, the numerical simulation of the liquid nitrogen freezing-temperature field of a single pipe is conducted using the Abaqus finite-element program. Results show that the numerical simulation of soil temperature distribution law well agrees with the analytical solution, further verifies the reliability of the established analytical solution of the liquid nitrogen freezing-temperature field of a single pipe.

Macromolecular differentiation of Golgi stacks in root tips of Arabidopsis and Nicotiana seedlings as visualized in high pressure frozen and freeze-substituted samples

NASA Technical Reports Server (NTRS)

Staehelin, L. A.; Giddings, T. H. Jr; Kiss, J. Z.; Sack, F. D.

1990-01-01

The plant root tip represents a fascinating model system for studying changes in Golgi stack architecture associated with the developmental progression of meristematic cells to gravity sensing columella cells, and finally to "young" and "old", polysaccharide-slime secreting peripheral cells. To this end we have used high pressure freezing in conjunction with freeze-substitution techniques to follow developmental changes in the macromolecular organization of Golgi stacks in root tips of Arabidopsis and Nicotiana. Due to the much improved structural preservation of all cells under investigation, our electron micrographs reveal both several novel structural features common to all Golgi stacks, as well as characteristic differences in morphology between Golgi stacks of different cell types. Common to all Golgi stacks are clear and discrete differences in staining patterns and width of cis, medial and trans cisternae. Cis cisternae have the widest lumina (approximately 30 nm) and are the least stained. Medial cisternae are narrower (approximately 20 nm) and filled with more darkly staining products. Most trans cisternae possess a completely collapsed lumen in their central domain, giving rise to a 4-6 nm wide dark line in cross-sectional views. Numerous vesicles associated with the cisternal margins carry a non-clathrin type of coat. A trans Golgi network with clathrin coated vesicles is associated with all Golgi stacks except those of old peripheral cells. It is easily distinguished from trans cisternae by its blebbing morphology and staining pattern. The zone of ribosome exclusion includes both the Golgi stack and the trans Golgi network. Intercisternal elements are located exclusively between trans cisternae of columella and peripheral cells, but not meristematic cells. In older peripheral cells only trans cisternae exhibit slime-related staining. Golgi stacks possessing intercisternal elements also contain parallel rows of freeze-fracture particles in their trans cisternal membranes. We propose that intercisternal elements serve as anchors of enzyme complexes involved in the synthesis of polysaccharide slime molecules to prevent the complexes from being dragged into the forming secretory vesicles by the very large slime molecules. In addition, we draw attention to the similarities in composition and apparent site of synthesis of xyloglucans and slime molecules.

Freezing induces a loss of freeze tolerance in an overwintering insect.

PubMed

Brown, C L; Bale, J S; Walters, K F A

2004-07-22

Cold-hardy insects overwinter by one of two main strategies: freeze tolerance and freeze avoidance by supercooling. As a general model, many freeze-tolerant species overwinter in extreme climates, freeze above -10 degrees C via induction by ice-nucleating agents, and once frozen, can survive at temperatures of up to 40 degrees C or more below the initial freezing temperature or supercooling point (SCP). It has been assumed that the SCP of freeze-tolerant insects is unaffected by the freezing process and that the freeze-tolerant state is therefore retained in winter though successive freeze-thaw cycles of the body tissues and fluids. Studies on the freeze-tolerant larva of the hoverfly Syrphus ribesii reveal this assumption to be untrue. When a sample with a mean 'first freeze' SCP of -7.6 degrees C (range of -5 degrees C to -9.5 degrees C) were cooled, either to -10 degrees C or to their individual SCP, on five occasions, the mean SCP was significantly depressed, with some larvae subsequently freezing as low as -28 degrees C. Only larvae that froze at the same consistently high temperature above -10 degrees C were alive after being frozen five times. The wider occurrence of this phenomenon would require a fundamental reassessment of the dynamics and distinctions of the freeze-tolerant and freeze-avoiding strategies of insect overwintering. Copyright 2004 The Royal Society

Repeated vitrification/warming of human sperm gives better results than repeated slow programmable freezing

PubMed Central

Vutyavanich, Teraporn; Lattiwongsakorn, Worashorn; Piromlertamorn, Waraporn; Samchimchom, Sudarat

2012-01-01

In this study, we compared the effects of repeated freezing/thawing of human sperm by our in-house method of rapid freezing with slow programmable freezing. Sperm samples from 11 normozoospermic subjects were processed through density gradients and divided into three aliquots: non-frozen, rapid freezing and slow programmable freezing. Sperm in the rapid freezing group had better motility and viability than those in the slow freezing group (P<0.01) after the first, second and third cycles of freezing/thawing, but there was no difference in morphology. In the second experiment, rapid freezing was repeated three times in 20 subjects. The samples from each thawing cycle were evaluated for DNA fragmentation using the alkaline comet assay. DNA fragmentation began to increase considerably after the second cycle of freezing/thawing, but to a level that was not clinically important. In the third experiment, rapid freezing was done repeatedly in 10 subjects, until no motile sperm were observed after thawing. The median number of repeated freezing/thawing that yielded no motile sperm was seven (range: 5–8, mean: 6.8). In conclusion, we demonstrated that repeated freezing/thawing of processed semen using our rapid freezing method gave better results than standard slow programmable freezing. This method can help maximize the usage of precious cryopreserved sperm samples in assisted reproduction technology. PMID:23064685

Characteristics of sugar surfactants in stabilizing proteins during freeze-thawing and freeze-drying.

PubMed

Imamura, Koreyoshi; Murai, Katsuyuki; Korehisa, Tamayo; Shimizu, Noriyuki; Yamahira, Ryo; Matsuura, Tsutashi; Tada, Hiroko; Imanaka, Hiroyuki; Ishida, Naoyuki; Nakanishi, Kazuhiro

2014-06-01

Sugar surfactants with different alkyl chain lengths and sugar head groups were compared for their protein-stabilizing effect during freeze-thawing and freeze-drying. Six enzymes, different in terms of tolerance against inactivation because of freeze-thawing and freeze-drying, were used as model proteins. The enzyme activities that remained after freeze-thawing and freeze-drying in the presence of a sugar surfactant were measured for different types and concentrations of sugar surfactants. Sugar surfactants stabilized all of the tested enzymes both during freeze-thawing and freeze-drying, and a one or two order higher amount of added sugar surfactant was required for achieving protein stabilization during freeze-drying than for the cryoprotection. The comprehensive comparison showed that the C10-C12 esters of sucrose or trehalose were the most effective through the freeze-drying process: the remaining enzyme activities after freeze-thawing and freeze-drying increased at the sugar ester concentrations of 1-10 and 10-100 μM, respectively, and increased to a greater extent than for the other surfactants at higher concentrations. Results also indicate that, when a decent amount of sugar was also added, the protein-stabilizing effect of a small amount of sugar ester through the freeze-drying process could be enhanced. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

Ice crystallization in porous building materials: assessing damage using real-time 3D monitoring

NASA Astrophysics Data System (ADS)

Deprez, Maxim; De Kock, Tim; De Schutter, Geert; Cnudde, Veerle

2017-04-01

Frost action is one of the main causes of deterioration of porous building materials in regions at middle to high latitudes. Damage will occur when the internal stresses due to ice formation become larger than the strength of the material. Hence, the sensitivity of the material to frost damage is partly defined by the structure of the solid body. On the other hand, the size, shape and interconnection of pores manages the water distribution in the building material and, therefore, the characteristics of the pore space control potential to form ice crystals (Ruedrich et al., 2011). In order to assess the damage to building materials by ice crystallization, lot of effort was put into identifying the mechanisms behind the stress build up. First of all, volumetric expansion of 9% (Hirschwald, 1908) during the transition of water to ice should be mentioned. Under natural circumstances, however, water saturation degrees within natural rocks or concrete cannot reach a damaging value. Therefore, linear growth pressure (Scherer, 1999), as well as several mechanisms triggered by water redistribution during freezing (Powers and Helmuth, 1953; Everett, 1961) are more likely responsible for damage due to freezing. Nevertheless, these theories are based on indirect observations and models and, thus, direct evidence that reveals the exact damage mechanism under certain conditions is still lacking. To obtain this proof, in-situ information needs to be acquired while a freezing process is performed. X-ray computed tomography has proven to be of great value in material research. Recent advances at the Ghent University Centre for Tomography (UGCT) have already allowed to dynamically 3D image crack growth in natural rock during freeze-thaw cycles (De Kock et al., 2015). A great potential to evaluate the different stress build-up mechanisms can be found in this imaging technique consequently. It is required to cover a range of materials with different petrophysical properties to achieve a correct assessment of the acting damage processes. Natural building stones tend to have an accessible open pore structure, whereas concrete is significantly less permeable and has large gel pores. Three sedimentary rocks (Bentheimer sandstone, Savonnières limestone and a classic Turkish travertine) and two types of concrete with a different water-cement ratio (0.6 and 0.5) are included in this research. With X-ray CT two approaches can be considered. First, an attempt directly to monitor the transition of water into ice can be performed. Secondly, the location of the induced micro-cracks within the pores or pore throats can possibly indicate which mechanism has caused them. Moreover, the retrieved CT data will be compared to different proxies for ice formation, such as length and temperature changes during freezing. This research is supported by FWO, project 3G004115. References: De Kock, T., et al. (2015). A Pore-Scale Study of Fracture Dynamics in Rock Using X-ray Micro- CT Under Ambient Freeze-Thaw Cycling. Environ. Sci. Technol. 49, 2867-2874. Everett, D.H. (1961). The thermodynamics of frost damage to porous solids. Trans. Faraday Soc. 57, 1541. Hirschwald, J. (1908). Die Prüfung der natürlichen Bausteine auf ihre Verwitterungsbeständigkeit. Verlag Wilhelm Ernst & Sohn, Berlin. Powers, T., Hemuth, R. (1953). Theory of volume chances in hardened portland-cement paste during freezing. Highway Research Board Proceedings, 32. Scherer, G. W. (1999). Crystallization in pores. Cem. Concr. Res., 29, 1347-1358. Ruedrich, J., Kirchner, D., Siegesmund, S. (2011). Physical weathering of building stones induced by freeze-thaw action: a laboratory long-term study. Environmental Earth Sciences, 63, 1573-1586.

The ability of the Antarctic nematode Panagrolaimus davidi to survive intracellular freezing is dependent upon nutritional status.

PubMed

Raymond, Mélianie R; Wharton, David A

2013-02-01

The Antarctic nematode Panagrolaimus davidi is the best documented example of an animal surviving intracellular freezing and the only animal so far shown to survive such freezing throughout its tissues. However, a recent study found that after exposure to a freezing stress that produced intracellular freezing in a proportion of nematodes, the resulting survival levels could be explained if those nematodes that froze intracellularly had died. We have thus re-examined the survival of intracellular freezing in this nematode. The ability to survive a freezing exposure that is likely to produce intracellular freezing (freezing at -10 °C) declines with culture age. In cultures that are fed regularly, the ability to survive freezing at -10 °C increases, but in starved cultures freezing survival declines. Survival of intracellular freezing in fed cultures was confirmed using cryomicroscopy, staining of cells with vital dyes and by freeze substitution and transmission electron microscopy. We have thus confirmed that P. davidi can survive intracellular freezing and shown that this ability is dependent upon them being well fed. The effect of culture conditions on the nutrient status of the nematodes should thus be an important factor in the design of experiments.

Emerging techniques for assisting and accelerating food freezing processes: A review of recent research progresses.

PubMed

Cheng, Lina; Sun, Da-Wen; Zhu, Zhiwei; Zhang, Zi

2017-03-04

Freezing plays an important role in food preservation and the emergence of rapid freezing technologies can be highly beneficial to the food industry. This paper reviews some novel food freezing technologies, including high-pressure freezing (HPF), ultrasound-assisted freezing (UAF), electrically disturbed freezing (EF) and magnetically disturbed freezing (MF), microwave-assisted freezing (MWF), and osmo-dehydro-freezing (ODF). HPF and UAF can initiate ice nucleation rapidly, leading to uniform distribution of ice crystals and the control of their size and shape. Specifically, the former is focused on increasing the degree of supercooling, whereas the latter aims to decrease it. Direct current electric freezing (DC-EF) and alternating current electric freezing (AC-EF) exhibit different effects on ice nucleation. DC-EF can promote ice nucleation and AC-EF has the opposite effect. Furthermore, ODF has been successfully used for freezing various vegetables and fruit. MWF cannot control the nucleation temperature, but can decrease supercooling degree, thus decreasing the size of ice crystals. The heat and mass transfer processes during ODF have been investigated experimentally and modeled mathematically. More studies should be carried out to understand the effects of these technologies on food freezing process.

Freezing and thawing or freezing, thawing, and aging effects on beef tenderness.

PubMed

Grayson, A L; King, D A; Shackelford, S D; Koohmaraie, M; Wheeler, T L

2014-06-01

The objective of this study was to determine the effect of freezing and thawing or freezing and thawing with an additional aging period after frozen storage on the tenderness of longissimus lumborum (LL) and semitendinosus (ST) steaks relative to aged, fresh steaks. Left-side LL and ST (n = 35 each) were obtained from U.S. Select carcasses classified at the grading stand by the U.S. Meat Animal Research Center visible and near-infrared spectroscopy tenderness system to have predicted slice shear force greater than 16.5 kg at 14 d postmortem. At 2 d postmortem, 2.54 cm thick steaks were cut from each muscle and assigned to 1 of the following treatments: 2 d fresh (2FRESH), 2 d freeze + thaw (2FREEZE), 2 d freeze + thaw + 12 d age (2FREEZE+12AGE), 14 d fresh (14FRESH), 14 d freeze + thaw (14FREEZE), 14 d freeze + thaw + 14 d age (14FREEZE+14AGE), and 28 d fresh (28FRESH). Steaks assigned to a freezing treatment were frozen at -26°C for 30 d before thawing/cooking or thawing with an additional aging period at 2°C. Slice shear force for LL and ST was lower (P < 0.01) for 2FREEZE (27.4 and 24.5 kg) and 14FREEZE (22.4 and 22.4 kg) compared to 2FRESH (33.0 and 29.2 kg) and 14FRESH (25.3 and 25.5 kg), respectively. Slice shear force for LL and ST was lower (P < 0.01) for 2FREEZE+12AGE (17.8 and 20.8 kg) and 14FREEZE+14AGE (14.6 and 19.0 kg) compared to 14FRESH (25.3 and 25.5 kg) and 28FRESH (18.7 and 21.7 kg), respectively. Desmin degradation for LL was not different (P > 0.05) between 2FREEZE (21.0%) and 2FRESH (14.6%) or between 14FREEZE (40.4%) and 14FRESH (38.4%); however, desmin degradation was higher (P < 0.06) in 2FREEZE+12AGE (46.7%) and 14FREEZE+14AGE (71.1%) when compared to 14FRESH (38.4%) and 28FRESH (60.5%), respectively. Cooking loss for LL was higher (P < 0.01) in 2FREEZE+12AGE (15.2%) compared to 14FRESH (14.0%) but was not different (P > 0.05) between 14FREEZE+14AGE (15.0%) and 28FRESH (14.3%). Freezing and thawing or a combination of freezing, thawing, and aging resulted in increased tenderness for LL and ST steaks when compared to fresh steaks with the same aging time. These results indicate freezing could be incorporated into normal commercial product distribution processes to improve the consistency of meat tenderness. Researchers who freeze steaks before tenderness assessment should be aware and acknowledge that freezing affects tenderness data.

Modeling Mantle Shear Zones, Melt Focusing and Stagnation - Are Non Volcanic Margins Really Magma Poor?

NASA Astrophysics Data System (ADS)

Lavier, L. L.; Muntener, O.

2011-12-01

Mantle peridotites from ocean-continent transition zones (OCT's) and ultraslow spreading ridges question the commonly held assumption of a simple link between mantle melting and MORB. 'Ancient' and partly refertilized mantle in rifts and ridges illustrates the distribution of the scale of upper mantle heterogeneity even on a local scale. Upwelling of partial melts that enter the conductive lithospheric mantle inevitably leads to freezing of the melt and metasomatized lithosphere. Field data and petrology demonstrates that ancient, thermally undisturbed, pyroxenite-veined subcontinental mantle blobs formed parts of the ocean floor next to thinned continental crust. Similar heterogeneity might be created in the oceanic lithosphere where the thermal boundary layer (TBM) is thick and veined with metasomatic assemblages. This cold, ancient, 'subcontinental domain' is separated by ductile shear zones (or some other form of permeability barriers) from an infiltrated ('hot') domain dominated by refertilized spinel and/or plagioclase peridotite. The footwall of these mantle shear zones display complex refertilization processes and high-temperature deformation. We present numerical models that illustrate the complex interplay of km-scale refertilization with active deformation and melt focusing on top of the mantle. Melt lubricated shear zones focus melt flow in shear fractures (melt bands) occurring along grain boundaries. Continuous uplift and cooling leads to crystallization, and crystal plastic deformation prevails in the subsolidus state. Below 800oC if water is present deformation by shearing of phyllosilicates may become prevalent. We develop physical boundary conditions for which stagnant melt beneath a permeability barrier remains trapped rather than being extracted to the surface via melt-filled fractures. We explore the parameter space for fracturing and drainage and development of anastomozing impermeable shear zones. Our models might be useful to constrain the conditions and enigmatic development of magma-poor and magma rich margins.

Cognitive Factors Affecting Freeze-like Behavior in Humans.

PubMed

Alban, Michael W; Pocknell, Victoria

2017-01-01

Contemporary research on survival-related defensive behaviors has identified physiological markers of freeze/flight/fight. Our research focused on cognitive factors associated with freeze-like behavior in humans. Study 1 tested if an explicit decision to freeze is associated with the psychophysiological state of freezing. Heart rate deceleration occurred when participants chose to freeze. Study 2 varied the efficacy of freezing relative to other defense options and found "freeze" was responsive to variations in the perceived effectiveness of alternative actions. Study 3 tested if individual differences in motivational orientation affect preference for a "freeze" option when the efficacy of options is held constant. A trend in the predicted direction suggested that naturally occurring cognitions led loss-avoiders to select "freeze" more often than reward-seekers. In combination, our attention to the cognitive factors affecting freeze-like behavior in humans represents a preliminary step in addressing an important but neglected research area.

Response of seasonal soil freeze depth to climate change across China

NASA Astrophysics Data System (ADS)

Peng, Xiaoqing; Zhang, Tingjun; Frauenfeld, Oliver W.; Wang, Kang; Cao, Bin; Zhong, Xinyue; Su, Hang; Mu, Cuicui

2017-05-01

The response of seasonal soil freeze depth to climate change has repercussions for the surface energy and water balance, ecosystems, the carbon cycle, and soil nutrient exchange. Despite its importance, the response of soil freeze depth to climate change is largely unknown. This study employs the Stefan solution and observations from 845 meteorological stations to investigate the response of variations in soil freeze depth to climate change across China. Observations include daily air temperatures, daily soil temperatures at various depths, mean monthly gridded air temperatures, and the normalized difference vegetation index. Results show that soil freeze depth decreased significantly at a rate of -0.18 ± 0.03 cm yr-1, resulting in a net decrease of 8.05 ± 1.5 cm over 1967-2012 across China. On the regional scale, soil freeze depth decreases varied between 0.0 and 0.4 cm yr-1 in most parts of China during 1950-2009. By investigating potential climatic and environmental driving factors of soil freeze depth variability, we find that mean annual air temperature and ground surface temperature, air thawing index, ground surface thawing index, and vegetation growth are all negatively associated with soil freeze depth. Changes in snow depth are not correlated with soil freeze depth. Air and ground surface freezing indices are positively correlated with soil freeze depth. Comparing these potential driving factors of soil freeze depth, we find that freezing index and vegetation growth are more strongly correlated with soil freeze depth, while snow depth is not significant. We conclude that air temperature increases are responsible for the decrease in seasonal freeze depth. These results are important for understanding the soil freeze-thaw dynamics and the impacts of soil freeze depth on ecosystem and hydrological process.

Mechanisms of deterioration of nutrients. [of freeze dried foods

NASA Technical Reports Server (NTRS)

Karel, M.; Flink, J. M.

1976-01-01

Methods which produce freeze dried foods of improved quality were examined with emphasis on storage stability. Specific topics discussed include: microstructure of freeze dried systems, investigation of structural changes in freeze dried systems, artificial food matrices, osmotic preconcentration to yield improved quality freeze dried fruits, and storage stability of osmotically preconcentrated freeze dried fruits.

Effect of freeze/thaw cycles on several biomarkers in urine from patients with kidney disease.

PubMed

Zhang, Yinan; Luo, Yi; Lu, Huijuan; Wang, Niansong; Shen, Yixie; Chen, Ruihua; Fang, Pingyan; Yu, Hong; Wang, Congrong; Jia, Weiping

2015-04-01

Urine samples were collected from eleven randomly selected patients with kidney disease, including diabetic nephropathy, chronic nephritis, and nephritic syndrome. Urine samples were treated with one of four protocols for freezing and thawing: freeze directly and thaw directly; freeze directly and thaw by temperature gradient; freeze by temperature gradient and thaw directly; and freeze by temperature gradient and thaw by temperature gradient. After one to six freeze/thaw cycles at -20°C or -80°C, different biomarkers showed differential stabilities. The concentrations of total protein, calcium, and potassium did not change significantly after five freeze/thaw cycles at either -20°C or -80°C. Albumin could only sustain three freeze/thaw cycles at -20°C before it started to degrade. We recommend that urine be stored at -80°C as albumin and the organic ions could sustain five and six freeze/thaw cycles, respectively, using the simple "direct freeze and direct thaw" protocol. Furthermore, in most cases, gradient freeze/thaw cycles are not necessary for urine sample storage.

The Iceland Deep Drilling Project (IDDP): Deep Fluid Sampling in Fractured Quartz, Reykjanes Geothermal System, Iceland

NASA Astrophysics Data System (ADS)

Seward, R. J.; Reed, M. H.; Grist, H. R.; Fridriksson, T.; Danielsen, P.; Thorhallsson, S.; Elders, W. A.; Fridleifsson, G. O.

2011-12-01

In July of 2011 a fluid inclusion tool (FIT) was deployed in well RN-17b of the Reykjanes geothermal system, Iceland, with the goal of sampling fluids in situ at the deepest feed point in the well. The tool consists of a perforated stainless steel pipe containing eight stainless steel mesh canisters, each loaded with 10mm-scale blocks of thermally fractured quartz. Except for one control canister, in each canister the fractured quartz blocks were surrounded by a different grain size of SiO¬2 glass that ranged in size from 10μm-scale glass wool to cm-scale glass shards. The FIT was left in the well on a wireline at a depth of 2768m and retrieved after three weeks. The fluid at 2768m depth is known from November 2010 well logs to have a temperature of about 330°C and pressure of 170 bars, a pressure ~40 bar too high for boiling at that temperature. After retrieval, quartz in all of the canisters contained liquid-dominated fluid inclusions, but their quantity and size differed by canister. Groups of inclusions occur in healed fractures and both healed and open fracture surfaces are visible within single quartz blocks. Measurements on a heating and cooling stage yield approximant inclusion homogenization temperatures of 332°C and freezing points of -2.0°C. These measurements and a pressure of 170 bars yield trapping temperatures of 335°C and a NaCl weight percent of 3.4, both of which match known values, thus verifying that the device trapped fluids as intended. In upcoming studies, these fluids will be analyzed using bulk methods and LA-ICP-MS on individual inclusions. The glass added to the quartz blocks in the canisters allowed the Reykjanes fluids to precipitate enough quartz to heal fractures and trap fluids despite the fluid undersaturation in quartz. Almost all of the glass that was added to the canisters, 27 to 66 grams in each (except glass wool), was consumed in the experiment. Remaining glass was in the non-mesh bottom caps of the canisters where fluid flux may have been minimal, indicating that most of the dissolved SiO2 was carried away with flowing fluid. This may explain why not all fractures were healed, as they were in our previous closed-system laboratory experiments. Upon recovery from the well, the FIT and the canister contents were covered in fine black particles, the greatest quantity by far occurring in canisters that had contained glass wool as the SiO2 source. Preliminary SEM-EDS analyses show that the particles contain silica, iron, magnesium, and small amounts of zinc sulfide. The precipitation of sulfides from the fluid sampled in the quartz fractures provides a valuable constraint on interpretation of the fluid inclusion compositions.

Assessing storage of stability and mercury reduction of freeze-dried Pseudomonas putida within different types of lyoprotectant

NASA Astrophysics Data System (ADS)

Azoddein, Abdul Aziz Mohd; Nuratri, Yana; Azli, Faten Ahada Mohd; Bustary, Ahmad Bazli

2017-12-01

Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose was able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage at 4 °C without vacuum. Polyethylene glycol (PEG) pre-treated freeze dried cells and broth pre-treated freeze dried cells after the freeze-dry process recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introducing freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage of 3 weeks was 17.91 %. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been grown in agar. Result from this study may be beneficial and useful as initial reference before commercialized freeze-dried P. putida.

Synchrotron X-Ray Visualisation of Ice Formation in Insects during Lethal and Non-Lethal Freezing

PubMed Central

Sinclair, Brent J.; Gibbs, Allen G.; Lee, Wah-Keat; Rajamohan, Arun; Roberts, Stephen P.; Socha, John J.

2009-01-01

Although the biochemical correlates of freeze tolerance in insects are becoming well-known, the process of ice formation in vivo is subject to speculation. We used synchrotron x-rays to directly visualise real-time ice formation at 3.3 Hz in intact insects. We observed freezing in diapausing 3rd instar larvae of Chymomyza amoena (Diptera: Drosophilidae), which survive freezing if it occurs above −14°C, and non-diapausing 3rd instar larvae of C. amoena and Drosophila melanogaster (Diptera: Drosophilidae), neither of which survive freezing. Freezing was readily observed in all larvae, and on one occasion the gut was seen to freeze separately from the haemocoel. There were no apparent qualitative differences in ice formation between freeze tolerant and non-freeze tolerant larvae. The time to complete freezing was positively related to temperature of nucleation (supercooling point, SCP), and SCP declined with decreasing body size, although this relationship was less strong in diapausing C. amoena. Nucleation generally occurred at a contact point with the thermocouple or chamber wall in non-diapausing larvae, but at random in diapausing larvae, suggesting that the latter have some control over ice nucleation. There were no apparent differences between freeze tolerant and non-freeze tolerant larvae in tracheal displacement or distension of the body during freezing, although there was markedly more distension in D. melanogaster than in C. amoena regardless of diapause state. We conclude that although control of ice nucleation appears to be important in freeze tolerant individuals, the physical ice formation process itself does not differ among larvae that can and cannot survive freezing. This suggests that a focus on cellular and biochemical mechanisms is appropriate and may reveal the primary adaptations allowing freeze tolerance in insects. PMID:20011523

Using Power Ultrasound to Accelerate Food Freezing Processes: Effects on Freezing Efficiency and Food Microstructure.

PubMed

Zhang, Peizhi; Zhu, Zhiwei; Sun, Da-Wen

2018-05-31

Freezing is an effective way of food preservation. However, traditional freezing methods have the disadvantages of low freezing efficiency and generation of large ice crystals, leading to possible damage of food quality. Power ultrasound assisted freezing as a novel technique can effectively reduce the adverse effects during freezing process. This paper gives an overview on recent researches of power ultrasound technique to accelerate the food freezing processes and illustrates the main principles of power ultrasound assisted freezing. The effects of power ultrasound on liquid food, model solid food as well as fruit and vegetables are discussed, respectively, from the aspects of increasing freezing rate and improving microstructure. It is shown that ultrasound assisted freezing can effectively improve the freezing efficiency and promote the formation of small and evenly distributed ice crystals, resulting in better food quality. Different inherent properties of food samples affect the effectiveness of ultrasound application and optimum ultrasound parameters depend on the nature of the samples. The application of ultrasound to the food industry is more likely on certain types of food products and more efforts are still needed to realize the industrial translation of laboratory results.

Aquaporin-Based Biomimetic Polymeric Membranes: Approaches and Challenges

PubMed Central

Habel, Joachim; Hansen, Michael; Kynde, Søren; Larsen, Nanna; Midtgaard, Søren Roi; Jensen, Grethe Vestergaard; Bomholt, Julie; Ogbonna, Anayo; Almdal, Kristoffer; Schulz, Alexander; Hélix-Nielsen, Claus

2015-01-01

In recent years, aquaporin biomimetic membranes (ABMs) for water separation have gained considerable interest. Although the first ABMs are commercially available, there are still many challenges associated with further ABM development. Here, we discuss the interplay of the main components of ABMs: aquaporin proteins (AQPs), block copolymers for AQP reconstitution, and polymer-based supporting structures. First, we briefly cover challenges and review recent developments in understanding the interplay between AQP and block copolymers. Second, we review some experimental characterization methods for investigating AQP incorporation including freeze-fracture transmission electron microscopy, fluorescence correlation spectroscopy, stopped-flow light scattering, and small-angle X-ray scattering. Third, we focus on recent efforts in embedding reconstituted AQPs in membrane designs that are based on conventional thin film interfacial polymerization techniques. Finally, we describe some new developments in interfacial polymerization using polyhedral oligomeric silsesquioxane cages for increasing the physical and chemical durability of thin film composite membranes. PMID:26264033

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kononchik, Joseph P.; Vancini, Ricardo; Brown, Dennis T., E-mail: dennis_brown@ncsu.edu

Sindbis Virus (SV), the prototype alphavirus in the family togaviridae, infects both mammalian and insect cells. The ability of SV to infect cells possessing significantly different biochemical environments suggests that there may be a common mode of entry into each cell type. Previous studies show that up to 4 h post infection cells are permeable to small ions and alpha sarcin suggesting that the plasma membrane is compromised as infection takes place. Thin-section electron microscopy has also shown SV to bind to the plasma membrane and lose its electron dense core through a pore like structure developed upon interaction ofmore » the virus with the cell surface. Using freeze-fracture replicas, thin-sections and antibody labeling the data presented herein show virus associated with intramembrane particles on mosquito cells. These data suggest that the intramembrane particles associated with SV may be part of the pore structure consisting of virus proteins and cell receptor.« less

Niemann-Pick type C proteins promote microautophagy by expanding raft-like membrane domains in the yeast vacuole

PubMed Central

Tsuji, Takuma; Fujimoto, Megumi; Tatematsu, Tsuyako; Cheng, Jinglei; Orii, Minami; Takatori, Sho; Fujimoto, Toyoshi

2017-01-01

Niemann-Pick type C is a storage disease caused by dysfunction of NPC proteins, which transport cholesterol from the lumen of lysosomes to the limiting membrane of that compartment. Using freeze fracture electron microscopy, we show here that the yeast NPC orthologs, Ncr1p and Npc2p, are essential for formation and expansion of raft-like domains in the vacuolar (lysosome) membrane, both in stationary phase and in acute nitrogen starvation. Moreover, the expanded raft-like domains engulf lipid droplets by a microautophagic mechanism. We also found that the multivesicular body pathway plays a crucial role in microautophagy in acute nitrogen starvation by delivering sterol to the vacuole. These data show that NPC proteins promote microautophagy in stationary phase and under nitrogen starvation conditions, likely by increasing sterol in the limiting membrane of the vacuole. DOI: http://dx.doi.org/10.7554/eLife.25960.001 PMID:28590904

Enhanced flexibility and electron-beam-controlled shape recovery in alumina-coated Au and Ag core-shell nanowires

NASA Astrophysics Data System (ADS)

Vlassov, Sergei; Polyakov, Boris; Vahtrus, Mikk; Mets, Magnus; Antsov, Mikk; Oras, Sven; Tarre, Aivar; Arroval, Tõnis; Lõhmus, Rünno; Aarik, Jaan

2017-12-01

The proper choice of coating materials and methods in core-shell nanowire (NW) engineering is crucial to assuring improved characteristics or even new functionalities of the resulting composite structures. In this paper, we have reported electron-beam-induced reversible elastic-to-plastic transition in Ag/Al2O3 and Au/Al2O3 NWs prepared by the coating of Ag and Au NWs with Al2O3 by low-temperature atomic layer deposition. The observed phenomenon enabled freezing the bent core-shell NW at any arbitrary curvature below the yield strength of the materials and later restoring its initially straight profile by irradiating the NW with electrons. In addition, we demonstrated that the coating efficiently protects the core material from fracture and plastic yield, allowing it to withstand significantly higher deformations and stresses in comparison to uncoated NW.

Polymer-dispersed liquid crystal elastomers

NASA Astrophysics Data System (ADS)

Rešetič, Andraž; Milavec, Jerneja; Zupančič, Blaž; Domenici, Valentina; Zalar, Boštjan

2016-10-01

The need for mechanical manipulation during the curing of conventional liquid crystal elastomers diminishes their applicability in the field of shape-programmable soft materials and future applications in additive manufacturing. Here we report on polymer-dispersed liquid crystal elastomers, novel composite materials that eliminate this difficulty. Their thermal shape memory anisotropy is imprinted by curing in external magnetic field, providing for conventional moulding of macroscopically sized soft, thermomechanically active elastic objects of general shapes. The binary soft-soft composition of isotropic elastomer matrix, filled with freeze-fracture-fabricated, oriented liquid crystal elastomer microparticles as colloidal inclusions, allows for fine-tuning of thermal morphing behaviour. This is accomplished by adjusting the concentration, spatial distribution and orientation of microparticles or using blends of microparticles with different thermomechanical characteristics. We demonstrate that any Gaussian thermomechanical deformation mode (bend, cup, saddle, left and right twist) of a planar sample, as well as beat-like actuation, is attainable with bilayer microparticle configurations.

Thylakoid membrane landscape in the sixties: a tribute to Andrew Benson.

PubMed

Anderson, Jan M

2007-05-01

Prior to the 1960s, the model for the molecular structure of cell membranes consisted of a lipid bilayer held in place by a thin film of electrostatically-associated protein stretched over the bilayer surface: (the Danielli-Davson-Robertson "unit membrane" model). Andrew Benson, an expert in the lipids of chloroplast thylakoid membranes, questioned the relevance of the unit membrane model for biological membranes, especially for thylakoid membranes, instead of emphasizing evidence in favour of hydrophobic interactions of membrane lipids within complementary hydrophobic regions of membrane-spanning proteins. With Elliot Weier, Benson postulated a remarkable subunit lipoprotein monolayer model for thylakoids. Following the advent of freeze fracture microscopy and the fluid lipid-protein mosaic model by Singer and Nicolson, the subunits, membrane-spanning integral proteins, span a dynamic lipid bilayer. Now that high resolution X-ray structures of photosystems I and II are being revealed, the seminal contribution of Andrew Benson can be appreciated.

Tunicamycin Prevents Cellulose Microfibril Formation in Oocystis solitaria.

PubMed

Quader, H

1984-07-01

The effect of tunicamycin (TM) on the development of the cell wall in Oocystis solitaria has been investigated. It was found that 10 micromolar TM completely stops the assembly of new microfibrils as observed at the ultrastructural level. During cell wall formation, freeze fracture replicas of the E-face of the plasma membrane reveal two major substructures: the terminal complexes (TC), paired and unpaired, and the microfibril imprints extending from unpaired TCs. In cells treated for 3 hours or longer with TM, the TCs are no longer visible, whereas microfibril imprints are still present. Because of the reported highly selective mode of action of TM, our results implicate a role for lipid-intermediates in cellulose synthesis in O. solitaria. It is assumed that TM prevents the formation of a glycoprotein which probably is a fundamental part of the TCs and may act as a primer for the assembly of the microfibrils.

Three-dimensional textures and defects of soft material layering revealed by thermal sublimation.

PubMed

Yoon, Dong Ki; Kim, Yun Ho; Kim, Dae Seok; Oh, Seong Dae; Smalyukh, Ivan I; Clark, Noel A; Jung, Hee-Tae

2013-11-26

Layering is found and exploited in a variety of soft material systems, ranging from complex macromolecular self-assemblies to block copolymer and small-molecule liquid crystals. Because the control of layer structure is required for applications and characterization, and because defects reveal key features of the symmetries of layered phases, a variety of techniques have been developed for the study of soft-layer structure and defects, including X-ray diffraction and visualization using optical transmission and fluorescence confocal polarizing microscopy, atomic force microscopy, and SEM and transmission electron microscopy, including freeze-fracture transmission electron microscopy. Here, it is shown that thermal sublimation can be usefully combined with such techniques to enable visualization of the 3D structure of soft materials. Sequential sublimation removes material in a stepwise fashion, leaving a remnant layer structure largely unchanged and viewable using SEM, as demonstrated here using a lamellar smectic liquid crystal.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sajgalik, P.; Sedlacek, J.; Lences, Z.

Densification of silicon carbide without any sintering aids by hot-pressing and rapid hot pressing was investigated. Full density (>99% t.d.) has been reached at 1850 °C, a temperature of at least 150-200 °C lower compared to the up to now known solid state sintered silicon carbide powders. Silicon carbide was freeze granulated and heat treated prior the densification. Furthermore, evolution of microstructure, mechanical properties and creep behavior were evaluated and compared to reference ceramics from as received silicon carbide powder as well as those of commercial one. Novel method results in dense ceramics with Vickers hardness and indentation fracture toughnessmore » of 29.0 GPa and 5.25 MPam 1/2, respectively. Moreover, the creep rate of 3.8 x 10 –9 s –1 at 1450 °C and the load of 100 MPa is comparable to the commercial α-SiC solid state sintered at 2150 °C.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fontana, Juan; Lopez-Iglesias, Carmen; Tzeng, Wen-Ping

Viral factories are complex structures in the infected cell where viruses compartmentalize their life cycle. Rubella virus (RUBV) assembles factories by recruitment of rough endoplasmic reticulum (RER), mitochondria and Golgi around modified lysosomes known as cytopathic vacuoles or CPVs. These organelles contain active replication complexes that transfer replicated RNA to assembly sites in Golgi membranes. We have studied the structure of RUBV factory in three dimensions by electron tomography and freeze-fracture. CPVs contain stacked membranes, rigid sheets, small vesicles and large vacuoles. These membranes are interconnected and in communication with the endocytic pathway since they incorporate endocytosed BSA-gold. RER andmore » CPVs are coupled through protein bridges and closely apposed membranes. Golgi vesicles attach to the CPVs but no tight contacts with mitochondria were detected. Immunogold labelling confirmed that the mitochondrial protein p32 is an abundant component around and inside CPVs where it could play important roles in factory activities.« less

Novel Real-Time Diagnosis of the Freezing Process Using an Ultrasonic Transducer

PubMed Central

Tseng, Yen-Hsiang; Cheng, Chin-Chi; Cheng, Hong-Ping; Lee, Dasheng

2015-01-01

The freezing stage governs several critical parameters of the freeze drying process and the quality of the resulting lyophilized products. This paper presents an integrated ultrasonic transducer (UT) in a stainless steel bottle and its application to real-time diagnostics of the water freezing process. The sensor was directly deposited onto the stainless steel bottle using a sol-gel spray technique. It could operate at temperature range from −100 to 400 °C and uses an ultrasonic pulse-echo technique. The progression of the freezing process, including water-in, freezing point and final phase change of water, were all clearly observed using ultrasound. The ultrasonic signals could indicate the three stages of the freezing process and evaluate the cooling and freezing periods under various processing conditions. The temperature was also adopted for evaluating the cooling and freezing periods. These periods increased with water volume and decreased with shelf temperature (i.e., speed of freezing). This study demonstrates the effectiveness of the ultrasonic sensor and technology for diagnosing and optimizing the process of water freezing to save energy. PMID:25946629

A comparison of freezing-damage during isochoric and isobaric freezing of the potato.

PubMed

Lyu, Chenang; Nastase, Gabriel; Ukpai, Gideon; Serban, Alexandru; Rubinsky, Boris

2017-01-01

Freezing is commonly used for food preservation. It is usually done under constant atmospheric pressure (isobaric). While extending the life of the produce, isobaric freezing has detrimental effects. It causes loss of food weight and changes in food quality. Using thermodynamic analysis, we have developed a theoretical model of the process of freezing in a constant volume system (isochoric). The mathematical model suggests that the detrimental effects associated with isobaric freezing may be reduced in an isochoric freezing system. To explore this hypothesis, we performed a preliminary study on the isochoric freezing of a produce with which our group has experience, the potato. Experiments were performed in an isochoric freezing device we designed. The device is robust and has no moving parts. For comparison, we used a geometrically identical isobaric freezing device. Following freezing and thawing, the samples were weighed, examined with colorimetry, and examined with microscopy. It was found that potatoes frozen to -5 °C in an isochoric system experienced no weight loss and limited enzymatic browning. In contrast the -5 °C isobaric frozen potato experienced substantial weight loss and substantial enzymatic browning. Microscopic analysis shows that the structural integrity of the potato is maintained after freezing in the isochoric system and impaired after freezing in the isobaric system. Tissue damage during isobaric freezing is caused by the increase in extracellular osmolality and the mechanical damage by ice crystals. Our thermodynamic analysis predicts that during isochoric freezing the intracellular osmolality remains comparable to the extracellular osmolality and that isochoric systems can be designed to eliminate the mechanical damage by ice. The results of this preliminary study seem to confirm the theoretical predictions. This is a preliminary exploratory study on isochoric freezing of food. We have shown that the quality of a food product preserved by isochoric freezing is better than the quality of food preserved to the same temperature in isobaric conditions. Obviously, more extensive research remains to be done to extend this study to lower freezing temperatures and other food items.

A comparison of freezing-damage during isochoric and isobaric freezing of the potato

PubMed Central

Serban, Alexandru; Rubinsky, Boris

2017-01-01

Background Freezing is commonly used for food preservation. It is usually done under constant atmospheric pressure (isobaric). While extending the life of the produce, isobaric freezing has detrimental effects. It causes loss of food weight and changes in food quality. Using thermodynamic analysis, we have developed a theoretical model of the process of freezing in a constant volume system (isochoric). The mathematical model suggests that the detrimental effects associated with isobaric freezing may be reduced in an isochoric freezing system. To explore this hypothesis, we performed a preliminary study on the isochoric freezing of a produce with which our group has experience, the potato. Method Experiments were performed in an isochoric freezing device we designed. The device is robust and has no moving parts. For comparison, we used a geometrically identical isobaric freezing device. Following freezing and thawing, the samples were weighed, examined with colorimetry, and examined with microscopy. Results It was found that potatoes frozen to −5 °C in an isochoric system experienced no weight loss and limited enzymatic browning. In contrast the −5 °C isobaric frozen potato experienced substantial weight loss and substantial enzymatic browning. Microscopic analysis shows that the structural integrity of the potato is maintained after freezing in the isochoric system and impaired after freezing in the isobaric system. Discussion Tissue damage during isobaric freezing is caused by the increase in extracellular osmolality and the mechanical damage by ice crystals. Our thermodynamic analysis predicts that during isochoric freezing the intracellular osmolality remains comparable to the extracellular osmolality and that isochoric systems can be designed to eliminate the mechanical damage by ice. The results of this preliminary study seem to confirm the theoretical predictions. Conclusion This is a preliminary exploratory study on isochoric freezing of food. We have shown that the quality of a food product preserved by isochoric freezing is better than the quality of food preserved to the same temperature in isobaric conditions. Obviously, more extensive research remains to be done to extend this study to lower freezing temperatures and other food items. PMID:28533970

Mechanisms of deterioration of nutrients. [improved quality of freeze-dried foods

NASA Technical Reports Server (NTRS)

Karel, M.; Flink, J. M.

1978-01-01

Methods for improving the quality of freeze-dried foods were investigated. Areas discussed include: (1) microstructure of freeze-dried systems, (2) structural changes in freeze-dried systems, (3) artificial food matrices, and (4) osmotic preconcentration to yield improved freeze-dried products.

Accurate prediction of collapse temperature using optical coherence tomography-based freeze-drying microscopy.

PubMed

Greco, Kristyn; Mujat, Mircea; Galbally-Kinney, Kristin L; Hammer, Daniel X; Ferguson, R Daniel; Iftimia, Nicusor; Mulhall, Phillip; Sharma, Puneet; Kessler, William J; Pikal, Michael J

2013-06-01

The objective of this study was to assess the feasibility of developing and applying a laboratory tool that can provide three-dimensional product structural information during freeze-drying and which can accurately characterize the collapse temperature (Tc ) of pharmaceutical formulations designed for freeze-drying. A single-vial freeze dryer coupled with optical coherence tomography freeze-drying microscopy (OCT-FDM) was developed to investigate the structure and Tc of formulations in pharmaceutically relevant products containers (i.e., freeze-drying in vials). OCT-FDM was used to measure the Tc and eutectic melt of three formulations in freeze-drying vials. The Tc as measured by OCT-FDM was found to be predictive of freeze-drying with a batch of vials in a conventional laboratory freeze dryer. The freeze-drying cycles developed using OCT-FDM data, as compared with traditional light transmission freeze-drying microscopy (LT-FDM), resulted in a significant reduction in primary drying time, which could result in a substantial reduction of manufacturing costs while maintaining product quality. OCT-FDM provides quantitative data to justify freeze-drying at temperatures higher than the Tc measured by LT-FDM and provides a reliable upper limit to setting a product temperature in primary drying. Copyright © 2013 Wiley Periodicals, Inc.

Freeze-drying of nanosuspensions, 1: freezing rate versus formulation design as critical factors to preserve the original particle size distribution.

PubMed

Beirowski, Jakob; Inghelbrecht, Sabine; Arien, Albertina; Gieseler, Henning

2011-05-01

It has been recently reported in the literature that using a fast freezing rate during freeze-drying of drug nanosuspensions is beneficial to preserve the original particle size distribution. All freezing rates studied were obtained by utilizing a custom-made apparatus and were then indirectly related to conventional vial freeze-drying. However, a standard freeze-dryer is only capable of achieving moderate freezing rates in the shelf fluid circulation system. Therefore, it was the purpose of the present study to evaluate the possibility to establish a typical freezing protocol applicable to a standard freeze-drying unit in combination with an adequate choice of cryoprotective excipients and steric stabilizers to preserve the original particle size distribution. Six different drug nanosuspensions containing itraconazole as a drug model were studied using freeze-thaw experiments and a full factorial design to reveal major factors for the stabilization of drug nanosuspensions and the corresponding interactions. In contrast to previous reports, the freezing regime showed no significant influence on preserving the original particle size distribution, suggesting that the concentrations of both the steric stabilizer and the cryoprotective agent are optimized. Moreover, it could be pinpointed that the combined effect of steric stabilizer and cryoprotectant clearly contribute to nanoparticle stability. Copyright © 2010 Wiley-Liss, Inc.

Freeze Technology for Nuclear Applications - 13590

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rostmark, Susanne C.; Knutsson, Sven; Lindberg, Maria

2013-07-01

Freezing of soil materials is a complicated process of a number of physical processes: - freezing of pore water in a thermal gradient, - cryogenic suction causing water migration and - ice formation expanding pores inducing frost heave. Structural changes due to increase of effective stress during freezing also take place. The over consolidation gives a powerful dewatering/drying effect and the freeze process causes separation of contaminates. Artificial ground freezing (AGF is a well established technique first practiced in south Wales, as early as 1862. AGF is mostly used to stabilize tunnels and excavations. During the last ten years underwatermore » applications of freeze technologies based on the AGF have been explored in Sweden. The technology can, and has been, used in many different steps in a remediation action. Freeze Sampling where undisturbed samples are removed in both soft and hard sediment/sludge, Freeze Dredging; retrieval of sediment with good precision and minimal redistribution, and Freeze Drying; volume reduction of contaminated sludge/sediment. The application of these technologies in a nuclear or radioactive environment provides several advantages. Sampling by freezing gives for example an advantage of an undisturbed sample taken at a specified depth, salvaging objects by freezing or removal of sludges is other applications of this, for the nuclear industry, novel technology. (authors)« less

Summer Freezing Resistance: A Critical Filter for Plant Community Assemblies in Mediterranean High Mountains

PubMed Central

Pescador, David S.; Sierra-Almeida, Ángela; Torres, Pablo J.; Escudero, Adrián

2016-01-01

Assessing freezing community response and whether freezing resistance is related to other functional traits is essential for understanding alpine community assemblages, particularly in Mediterranean environments where plants are exposed to freezing temperatures and summer droughts. Thus, we characterized the leaf freezing resistance of 42 plant species in 38 plots at Sierra de Guadarrama (Spain) by measuring their ice nucleation temperature, freezing point (FP), and low-temperature damage (LT50), as well as determining their freezing resistance mechanisms (i.e., tolerance or avoidance). The community response to freezing was estimated for each plot as community weighted means (CWMs) and functional diversity (FD), and we assessed their relative importance with altitude. We established the relationships between freezing resistance, growth forms, and four key plant functional traits (i.e., plant height, specific leaf area, leaf dry matter content (LDMC), and seed mass). There was a wide range of freezing resistance responses and more than in other alpine habitats. At the community level, the CWMs of FP and LT50 responded negatively to altitude, whereas the FD of both traits increased with altitude. The proportion of freezing-tolerant species also increased with altitude. The ranges of FP and LT50 varied among growth forms, and only leaf dry matter content was negatively correlated with freezing-resistance traits. Summer freezing events represent important abiotic filters for assemblies of Mediterranean high mountain communities, as suggested by the CWMs. However, a concomitant summer drought constraint may also explain the high freezing resistance of species that thrive in these areas and the lower FD of freezing resistance traits at lower altitudes. Leaves with high dry matter contents may maintain turgor at lower water potential and enhance drought tolerance in parallel to freezing resistance. This adaptation to drought seems to be a general prerequisite for plants found in xeric mountains. PMID:26941761

Survival mechanisms of vertebrate ectotherms at subfreezing temperatures: applications in cryomedicine.

PubMed

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

1995-03-01

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.

Hepatitis B vaccine freezing in the Indonesian cold chain: evidence and solutions.

PubMed

Nelson, Carib M; Wibisono, Hariadi; Purwanto, Hary; Mansyur, Isa; Moniaga, Vanda; Widjaya, Anton

2004-02-01

To document and characterize freezing temperatures in the Indonesian vaccine cold chain and to evaluate the feasibility of changes designed to reduce the occurrence of freezing. Data loggers were used to measure temperatures of shipments of hepatitis B vaccine from manufacturer to point of use. Baseline conditions and three intervention phases were monitored. During each of the intervention phases, vaccines were removed progressively from the standard 2-8 degrees C cold chain. Freezing temperatures were recorded in 75% of baseline shipments. The highest rates of freezing occurred during transport from province to district, storage in district-level ice-lined refrigerators, and storage in refrigerators in health centres. Interventions reduced freezing, without excessive heat exposure. Inadvertent freezing of freeze-sensitive vaccines is widespread in Indonesia. Simple strategies exist to reduce freezing - for example, selective transport and storage of vaccines at ambient temperatures. The use of vaccine vial monitors reduces the risk associated with heat-damaged vaccines in these scenarios. Policy changes that allow limited storage of freeze-sensitive vaccines at temperatures >2-8 degrees C would enable flexible vaccine distribution strategies that could reduce vaccine freezing, reduce costs, and increase capacity.

Repeated freeze-thaw cycles induce embolism in drought stressed conifers (Norway spruce, stone pine).

PubMed

Mayr, Stefan; Gruber, Andreas; Bauer, Helmut

2003-07-01

Freezing and thawing lead to xylem embolism when gas bubbles caused by ice formation expand during the thaw process. However, previous experimental studies indicated that conifers are resistant to freezing-induced embolism, unless xylem pressure becomes very negative during the freezing. In this study, we show that conifers experienced freezing-induced embolism when exposed to repeated freeze-thaw cycles and simultaneously to drought. Simulating conditions at the alpine timberline (128 days with freeze-thaw events and thawing rates of up to 9.5 K h(-1) in the xylem of exposed twigs during winter), young trees of Norway spruce [Picea abies (L.) Karst.] and stone pine (Pinus cembra L.) were exposed to 50 and 100 freeze-thaw cycles. This treatment caused a significant increase in embolism rates in drought-stressed samples. Upon 100 freeze-thaw cycles, vulnerability thresholds (50% loss of conductivity) were shifted 1.8 MPa (Norway spruce) and 0.8 MPa (stone pine) towards less negative water potentials. The results demonstrate that freeze-thaw cycles are a possible reason for winter-embolism in conifers observed in several field studies. Freezing-induced embolism may contribute to the altitudinal limits of conifers.

Freezing of gait in Parkinson's disease: the paradoxical interplay between gait and cognition.

PubMed

Ricciardi, Lucia; Bloem, Bastiaan R; Snijders, Anke H; Daniele, Antonio; Quaranta, Davide; Bentivoglio, Anna Rita; Fasano, Alfonso

2014-08-01

Freezing of gait is a disabling episodic gait disturbance common in patients with Parkinson's disease. Recent evidences suggest a complex interplay between gait impairment and executive functions. Aim of our study was to evaluate whether specific motor conditions (sitting or walking) influence cognitive performance in patients with or without different types of freezing. Eight healthy controls, eight patients without freezing, nine patients with levodopa-responsive and nine patients with levodopa-resistant freezing received a clinical and neuropsychological assessment during two randomly performed conditions: at rest and during walking. At rest, patients with levodopa-resistant freezing performed worse than patients without freezing on tests of phonological fluency (p = 0.01). No differences among the four groups were detected during walking. When cognitive performances during walking were compared to the performance at rest, there was a significant decline of verbal episodic memory task (Rey Auditory Verbal Learning Test) in patients without freezing and with levodopa-responsive freezing. Interestingly, walking improved performance on the phonological fluency task in patients with levodopa-resistant freezing (p = 0.04). Compared to patients without freezing, patients with levodopa-resistant freezing perform worse when tested while seated in tasks of phonological verbal fluency. Surprisingly, gait was associated with a paradoxical improvement of phonological verbal fluency in the patients with levodopa-resistant freezing whilst walking determined a worsening of episodic memory in the other patient groups. Copyright © 2014 Elsevier Ltd. All rights reserved.

Freezing during tapping tasks in patients with advanced Parkinson's disease and freezing of gait.

PubMed

Delval, Arnaud; Defebvre, Luc; Tard, Céline

2017-01-01

Parkinson's disease patients with freezing of gait also experience sudden motor blocks (freezing) during other repetitive motor tasks. We assessed the proportion of patients with advanced PD and freezing of gait who also displayed segmental "freezing" in tapping tasks. Fifteen Parkinson's disease patients with freezing of gait were assessed. Freezing of gait was evaluated using a standardized gait trajectory with the usual triggers. Patients performed repetitive tapping movements (as described in the MDS-UPDRS task) with the hands or the feet in the presence or absence of a metronome set to 4 Hz. Movements were recorded with a video motion system. The primary endpoint was the occurrence of segmental freezing in these tapping tasks. The secondary endpoints were (i) the relationship between segmental episodic phenomena and FoG severity, and (ii) the reliability of the measurements. For the upper limbs, freezing was observed more frequently with a metronome (21% of trials) than without a metronome (5%). For the lower limbs, the incidence of freezing was higher than for the upper limbs, and was again observed more frequently in the presence of an auditory cue (47%) than in its absence (14%). Although freezing of the lower limbs was easily assessed during an MDS-UPDRS task with a metronome, it was not correlated with the severity of freezing of gait (as evaluated during a standardized gait trajectory). Only this latter was a reliable measurement in patients with advanced Parkinson's disease.

Short-term and practice effects of metronome pacing in Parkinson's disease patients with gait freezing while in the 'on' state: randomized single blind evaluation.

PubMed

Cubo, Esther; Leurgans, Sue; Goetz, Christopher G

2004-12-01

In a randomized single blind parallel study, we tested the efficacy of an auditory metronome on walking speed and freezing in Parkinson's disease (PD) patients with freezing gait impairment during their 'on' function. No pharmacological treatment is effective in managing 'on' freezing in PD. Like visual cues that can help overcome freezing, rhythmic auditory pacing may provide cues that help normalize walking pace and overcome freezing. Non-demented PD patients with freezing during their 'on' state walked under two conditions, in randomized order: unassisted walking and walking with the use of an audiocassette with a metronome recording. The walking trials were randomized and gait variables were rated from videotapes by a blinded evaluator. Outcome measures were total walking time (total trial time-total freezing time), which was considered the time over a course of specified length, freezing time, average freeze duration and number of freezes. All outcomes were averaged across trials for each person and then compared across conditions using Signed Rank tests. Twelve non-demented PD patients with a mean age of 65.8 +/- 11.2 years, and mean PD duration of 12.4 +/- 7.3 years were included. The use of the metronome slowed ambulation and increased the total walking time (P < 0.0005) only during the first visit, without affecting any freezing variable. In the nine patients who took the metronome recording home and used it daily for 1 week while walking, freezing remained unimproved. Though advocated in prior publications as a walking aid for PD patients, auditory metronome pacing slows walking and is not a beneficial intervention for freezing during their 'on' periods.

Freezing Injury in Onion Bulb Cells

PubMed Central

Palta, Jiwan P.; Levitt, Jacob; Stadelmann, Eduard J.

1977-01-01

Onion (Allium cepa L.) bulbs were subjected for 12 days to either a moderate freeze (−4 C) or a severe freeze (−11 C). They were then thawed slowly over ice. During 7 to 12 days following the thaw, the injury progressed with time in the severely frozen bulbs, but appeared completely repaired in the moderately frozen bulbs. This was shown by the following post-thawing changes. Infiltration of the intercellular spaces increased from 80 to 90% to 100% after the severe freeze, and decreased from 30 to 50% to zero after the moderate freeze. All of the cells were alive immediately after thawing whether the freeze was moderate or severe. Corresponding to the infiltration results 7 to 12 days later, many to most were dead following the severe freeze, all were alive following the moderate freeze. The conductivity of the effusate from pieces of bulb tissue increased after the severe freezing, and decreased after the moderate freezing. The concentration of K+, total solutes, and sugars in the effusate paralleled the conductivity changes. Neither the pH of the effusate nor the permeability of the cells (as long as cells were living) to water was changed following either the severe or the moderate freezes. Some treatments of the thawed tissue following the severe freeze halted the progress of injury. The above results indicate that the semipermeable properties of the cell are uninjured but that the ion and sugar transport mechanism is damaged by freezing. Most likely the primary injury is to the active transport mechanism involved in their transport. It must be concluded that the final injury following freezing and thawing cannot be evaluated from the degree of infiltration or the conductivity of the effusate immediately after thawing, since injury may progress or recede following the thawing. PMID:16660101

Effects of Pressure-shift Freezing on the Structural and Physical Properties of Gelatin Hydrogel Matrices

PubMed Central

Kim, Byeongsoo; Gil, Hyung Bae; Min, Sang-Gi; Lee, Si-Kyung; Choi, Mi-Jung

2014-01-01

This study investigates the effects of the gelatin concentration (10-40%, w/v), freezing temperatures (from -20℃ to -50℃) and freezing methods on the structural and physical properties of gelatin matrices. To freeze gelatin, the pressure-shift freezing (PSF) is being applied at 0.1 (under atmospheric control), 50 and 100 MPa, respectively. The freezing point of gelatin solutions decrease with increasing gelatin concentrations, from -0.2℃ (10% gelatin) to -6.7℃ (40% gelatin), while the extent of supercooling did not show any specific trends. The rheological properties of the gelatin indicate that both the storage (G') and loss (G") moduli were steady in the strain amplitude range of 0.1-10%. To characterize gelatin matrices formed by the various freezing methods, the ice crystal sizes which were being determined by the scanning electron microscopy (SEM) are affected by the gelatin concentrations. The ice crystal sizes are affected by gelatin concentrations and freezing temperature, while the size distributions of ice crystals depend on the freezing methods. Smaller ice crystals are being formed with PSF rather than under the atmospheric control where the freezing temperature is above -40℃. Thus, the results of this study indicate that the PSF processing at a very low freezing temperature (-50℃) offers a potential advantage over commercial atmospheric freezing points for the formation of small ice crystals. PMID:26760743

Massive formation of square array junctions dramatically alters cell shape but does not cause lens opacity in the cav1-KO mice.

PubMed

Biswas, Sondip K; Brako, Lawrence; Lo, Woo-Kuen

2014-08-01

The wavy square array junctions are composed of truncated aquaporin-0 (AQP0) proteins typically distributed in the deep cortical and nuclear fibers in wild-type lenses. These junctions may help maintain the narrowed extracellular spaces between fiber cells to minimize light scattering. Herein, we investigate the impact of the cell shape changes, due to abnormal formation of extensive square array junctions, on the lens opacification in the caveolin-1 knockout mice. The cav1-KO and wild-type mice at age 1-22 months were used. By light microscopy examinations, cav1-KO lenses at age 1-18 months were transparent in both cortical and nuclear regions, whereas some lenses older than 18 months old exhibited nuclear cataracts. Scanning EM consistently observed the massive formation of ridge-and-valley membrane surfaces in young fibers at approximately 150 μm deep in all cav1-KO lenses studied. In contrast, the typical ridge-and-valleys were only seen in mature fibers deeper than 400 μm in wild-type lenses. The resulting extensive ridge-and-valleys dramatically altered the overall cell shape in cav1-KO lenses. Remarkably, despite dramatic shape changes, these deformed fiber cells remained intact and made close contact with their neighboring cells. By freeze-fracture TEM, ridge-and-valleys exhibited the typical orthogonal arrangement of 6.6 nm square array intramembrane particles and displayed the narrowed extracellular spaces. Immunofluorescence analysis showed that AQP0 C-terminus labeling was significantly decreased in outer cortical fibers in cav1-KO lenses. However, freeze-fracture immunogold labeling showed that the AQP0 C-terminus antibody was sparsely distributed on the wavy square array junctions, suggesting that the cleavage of AQP0 C-termini might not yet be complete. The cav1-KO lenses with nuclear cataracts showed complete cellular breakdown and large globule formation in the lens nucleus. This study suggests that despite dramatic cell shape changes, the massive formation of wavy square array junctions in intact fibers may provide additional adhesive support for maintaining the narrowed extracellular spaces that are crucial for the transparency of cav1-KO lenses. Copyright © 2014 Elsevier Ltd. All rights reserved.

Evaluation of combined effects of ageing period and freezing rate on quality attributes of beef loins.

PubMed

Kim, Yuan H Brad; Liesse, Charlotte; Kemp, Robert; Balan, Prabhu

2015-12-01

The objective of our study was to evaluate the combined effects of ageing period and different freezing rates on meat quality attributes of beef loins. Pairs of loins (M. longissimus at 1 day post mortem) from 12 carcasses were divided into four equal portions and randomly assigned to four ageing/freezing treatments (aged only, frozen only, and 3 or 4 weeks ageing at -1.5°C then frozen). Two freezing methods (fast freezing by calcium chloride immersion or slow freezing by air freezer at -18°C) were applied to the loin sections. Fast freezing had no effect on shear force (P>0.05), but significantly improved the water-holding capacity of the aged/frozen loins by reducing purge and drip losses. Ageing-then-freezing significantly improved shear force values of loins compared to both the aged only and frozen only loins. These observations suggest that fast freezing will add more value to the aged/frozen/thawed meat by minimising the amount of water-loss due to the freezing/thawing process. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effects of different freezing methods on calcium enriched papaya (Carica papaya L.).

PubMed

Lovera, Nancy N; Ramallo, Laura; Salvadori, Viviana O

2018-06-01

The effect of calcium impregnation on drip loss, colour, mechanical properties, sensory perception and freezing time on frozen-thawed papaya was studied, evaluating different freezing methods: cryogenic, tunnel and household freezer freezing. Osmotic dehydration as pre-treatment was also evaluated. Freezing in liquid nitrogen was considered an inappropriate method for papaya preservation due to cracking. Calcium impregnation and osmotic dehydration increased tissue firmness and decreased freezing time (freezing time for fresh, calcium impregnated and osmo-dehydrated fruit was 23, 17 and 5 min in a tunnel and 118, 83 and 60 min in a household freezer, respectively). Calcium lactate was the most effective way to protect tissue's firmness before and after a freeze-thaw cycle (maximum stress values approx. 300-400% of the raw tissue for tunnel freezing and 260% for household freezer). Microstructure analysis showed better tissue integrity retention in papaya samples impregnated with calcium lactate than in those with calcium gluconate, after a freezing-thawing cycle, in agreement with the drip loss results. In spite of these results, consumers preferred frozen papaya without pre-treatment or impregnated with calcium gluconate.

Response of shoal grass, Halodule wrightii, to extreme winter conditions in the Lower Laguna Madre, Texas

USGS Publications Warehouse

Hicks, D.W.; Onuf, C.P.; Tunnell, J.W.

1998-01-01

Effects of a severe freeze on the shoal grass, Halodule wrightii, were documented through analysis of temporal and spatial trends in below-ground biomass. The coincidence of the second lowest temperature (-10.6??C) in 107 years of record, 56 consecutive hours below freezing, high winds and extremely low water levels exposed the Laguna Madre, TX, to the most severe cold stress in over a century. H. wrightii tolerated this extreme freeze event. Annual pre- and post-freeze surveys indicated that below-ground biomass estimated from volume was Unaffected by the freeze event. Nor was there any post-freeze change in biomass among intertidal sites directly exposed to freezing air temperatures relative to subtidal sites which remained submerged during the freezing period.

The effect of undissolved air on isochoric freezing.

PubMed

Perez, Pedro A; Preciado, Jessica; Carlson, Gary; DeLonzor, Russ; Rubinsky, Boris

2016-06-01

This study evaluates the effect of undissolved air on isochoric freezing of aqueous solutions. Isochoric freezing is concerned with freezing in a constant volume thermodynamic system. A possible advantage of the process is that it substantially reduces the percentage of ice in the system at every subzero temperature, relative to atmospheric freezing. At the pressures generated by isochoric freezing, or high pressure isobaric freezing, air cannot be considered an incompressible substance and the presence of undissolved air substantially increases the amount of ice that forms at any subfreezing temperature. This effect is measurable at air volumes as low as 1%. Therefore eliminating the undissolved air, or any separate gaseous phase, from the system is essential for retaining the properties of isochoric freezing. Copyright © 2016. Published by Elsevier Inc.

Visual Indicators on Vaccine Boxes as Early Warning Tools to Identify Potential Freeze Damage.

PubMed

Angoff, Ronald; Wood, Jillian; Chernock, Maria C; Tipping, Diane

2015-07-01

The aim of this study was to determine whether the use of visual freeze indicators on vaccines would assist health care providers in identifying vaccines that may have been exposed to potentially damaging temperatures. Twenty-seven sites in Connecticut involved in the Vaccine for Children Program participated. In addition to standard procedures, visual freeze indicators (FREEZEmarker ® L; Temptime Corporation, Morris Plains, NJ) were affixed to each box of vaccine that required refrigeration but must not be frozen. Temperatures were monitored twice daily. During the 24 weeks, all 27 sites experienced triggered visual freeze indicator events in 40 of the 45 refrigerators. A total of 66 triggered freeze indicator events occurred in all 4 types of refrigerators used. Only 1 of the freeze events was identified by a temperature-monitoring device. Temperatures recorded on vaccine data logs before freeze indicator events were within the 35°F to 46°F (2°C to 8°C) range in all but 1 instance. A total of 46,954 doses of freeze-sensitive vaccine were stored at the time of a visual freeze indicator event. Triggered visual freeze indicators were found on boxes containing 6566 doses (14.0% of total doses). Of all doses stored, 14,323 doses (30.5%) were of highly freeze-sensitive vaccine; 1789 of these doses (12.5%) had triggered indicators on the boxes. Visual freeze indicators are useful in the early identification of freeze events involving vaccines. Consideration should be given to including these devices as a component of the temperature-monitoring system for vaccines.

Influence of the freezing method on the changes that occur in grape samples after frozen storage.

PubMed

Santesteban, Luis G; Miranda, Carlos; Royo, José B

2013-09-01

Sample freezing is frequently used in oenological laboratories as a compromise solution to increase the number of samples that can be analysed, despite the fact that some grape characteristics are known to change after frozen storage. However, freezing is usually performed using standard freezers, which provide a slow freezing. The aim of this work was to evaluate whether blast freezing would decrease the impact of standard freezing on grape composition. Grape quality parameters were assessed in fresh and in frozen stored samples that had been frozen using three different procedures: standard freezing and blast freezing using either a blast freezer or an ultra-freezer. The implications of frozen storage in grape samples reported in earlier research were observed for the three freezing methods evaluated. Although blast freezing improved repeatability for the most problematic parameters (tartaric acidity, TarA; total phenolics, TP), the improvement was not important from a practical point of view. However, TarA and TP were relatively repeatable among the three freezing procedures, which suggests that freezing had an effect on these parameters independently of the method used . According to our results, the salification potential of the must is probably implied in the changes observed for TarA, whereas for TP the precipitation of protoanthocyanins after association with cell wall material is hypothesized to cause the lack of repeatability between fresh and frozen grapes. Blast freezing would not imply a great improvement if implemented in oenological laboratories, at least for the parameters included in this study. © 2013 Society of Chemical Industry.

The Adsorption and Desorption of Pb(2+) and Cd(2+) in Freeze-Thaw Treated Soils.

PubMed

Li, Linhui; Ma, Jincai; Xu, Meng; Li, Xu; Tao, Jiahui; Wang, Guanzhu; Yu, Jitong; Guo, Ping

2016-01-01

Adsorption and desorption are important processes that influence the potential toxicity and bioavailability of heavy metals in soils. However, information regarding adsorption and desorption behavior of heavy metals in soils subjected to freeze-thaw cycles is poorly understood. In the current study, the effect of freeze-thaw cycles with different freezing temperature (-15, -25, -35°C) on soil properties was investigated. Then the adsorption and desorption behavior of Pb(2+) and Cd(2+) in freeze-thaw treated soils was studied. The adsorption amounts of Pb(2+) and Cd(2+) in freeze-thaw treated soils were smaller than those in unfrozen soils (p < 0.05), due to the fact that pH, cation exchange capacity, organic matter content, free iron oxide content, and CaCO3 content in freeze-thaw treated soils were smaller than those in unfrozen soils. The adsorption amounts of Pb(2+) and Cd(2+) in soils treated with lower freezing temperatures were higher than those in soils treated with higher freezing temperatures. Desorption percentages of Pb(2+) and Cd(2+) in unfrozen soils were smaller than those in freeze-thaw treated soils (p < 0.05). The desorption percentages of Pb(2+) and Cd(2+) were smaller in soils treated with lower freezing temperatures than those in soils treated with higher freezing temperatures. The results obtained highlight the change of the adsorption and desorption behavior of typical heavy metals in freeze-thaw treated soils located in seasonal frozen soils zone in northeast China.

Evaluating Long-Term Impacts of Soil-Mixing Source-Zone Treatment using Cryogenic Core Collection

DTIC Science & Technology

2017-06-01

to (a) coring equipment freezing downhole, (b) freezing or binding of the core sample in barrel, and ( c ) running out of LN in the vicinity of sampling...encountered due to (a) coring equipment freezing downhole, (b) freezing or binding of the core sample in barrel, and ( c ) running out of LN in the...equipment freezing downhole, (b) freezing or binding of the core sample in barrel, and ( c ) running out of LN in the vicinity of sampling. Downhole

Insufficiency of copper ion homeostasis causes freeze-thaw injury of yeast cells as revealed by indirect gene expression analysis.

PubMed

Takahashi, Shunsuke; Ando, Akira; Takagi, Hiroshi; Shima, Jun

2009-11-01

Saccharomyces cerevisiae is exposed to freeze-thaw stress in commercial processes, including frozen dough baking. Cell viability and fermentation activity after a freeze-thaw cycle were dramatically decreased due to freeze-thaw injury. Because this type of injury involves complex phenomena, the injury mechanisms are not fully understood. We examined freeze-thaw injury by indirect gene expression analysis during postthaw incubation after freeze-thaw treatment using DNA microarray profiling. The results showed that genes involved in the homeostasis of metal ions were frequently contained in genes that were upregulated, depending on the freezing period. We assessed the phenotype of deletion mutants of the metal ion homeostasis genes that exhibited freezing period-dependent upregulation and found that the strains with deletion of the MAC1 and CTR1 genes involved in copper ion homeostasis exhibited freeze-thaw sensitivity, suggesting that copper ion homeostasis is required for freeze-thaw tolerance. We found that supplementation with copper ions during postthaw incubation increased intracellular superoxide dismutase activity and intracellular levels of reactive oxygen species were decreased. Moreover, cell viability was increased by supplementation with copper ions. These results suggest that insufficiency of copper ion homeostasis may be one of the causes of freeze-thaw injury.

Application of the Quality by Design Approach to the Freezing Step of Freeze-Drying: Building the Design Space.

PubMed

Arsiccio, Andrea; Pisano, Roberto

2018-06-01

The present work shows a rational method for the development of the freezing step of a freeze-drying cycle. The current approach to the selection of freezing conditions is still empirical and nonsystematic, thus resulting in poor robustness of control strategy. The final aim of this work is to fill this gap, describing a rational procedure, based on mathematical modeling, for properly choosing the freezing conditions. Mechanistic models are used for the prediction of temperature profiles during freezing and dimension of ice crystals being formed. Mathematical description of the drying phase of freeze-drying is also coupled with the results obtained by freezing models, thus providing a comprehensive characterization of the lyophilization process. In this framework, deep understanding of the phenomena involved is required, and according to the Quality by Design approach, this knowledge can be used to build the design space. The step-by-step procedure for building the design space for freezing is thus described, and examples of applications are provided. The calculated design space is validated upon experimental data, and we show that it allows easy control of the freezing process and fast selection of appropriate operating conditions. Copyright © 2018 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Freezing curve-based monitoring to quickly evaluate the viability of biological materials subject to freezing or thermal injury.

PubMed

Liu, Jing; Zhou, Yi-Xin

2003-09-01

This paper is aimed at investigating the roles of freezing dynamics of a liquid droplet to characterize the properties of the material. In particular, freezing curve-based monitoring was proposed to quickly evaluate the viability of biological materials subject to freezing, re-warming, or other kinds of injury, which is an extremely important issue in practices such as cryobiology, hyperthermia, or freshness evaluation of bio-samples. An integrated micro analysis device was fabricated which is simple in structure and cheap to make. Preliminary freezing results demonstrated that minor changes in a biological material due to freezing or warming injury might result in a significant deviation of its freezing curve from that of the intact biomaterials. Several potential thermal indexes to quantify the material features were pointed out. Further, experiments were performed on some freezing and thawing processes of small amount of water on a cooling surface to test the effects of droplet sizes, measurement sites, cooling strength, and cooling geometry, etc., on the freezing responses of a water droplet. Their implementation in developing a new micro analysis system were suggested. This freezing curve-based monitoring method may open a new strategy for the evaluation of biomaterials subject to destruction in diverse fields.

Recent developments in smart freezing technology applied to fresh foods.

PubMed

Xu, Ji-Cheng; Zhang, Min; Mujumdar, Arun S; Adhikari, Benu

2017-09-02

Due to the increased awareness of consumers in sensorial and nutritional quality of frozen foods, the freezing technology has to seek new and innovative technologies for better retaining the fresh like quality of foods. In this article, we review the recent developments in smart freezing technology applied to fresh foods. The application of these intelligent technologies and the associated underpinning concepts have greatly improved the quality of frozen foods and the freezing efficiency. These technologies are able to automatically collect the information in-line during freezing and help control the freezing process better. Smart freezing technology includes new and intelligent technologies and concepts applied to the pretreatment of the frozen product, freezing processes, cold chain logistics as well as warehouse management. These technologies enable real-time monitoring of quality during the freezing process and help improve product quality and freezing efficiency. We also provide a brief overview of several sensing technologies used to achieve automatic control of individual steps of freezing process. These sensing technologies include computer vision, electronic nose, electronic tongue, digital simulation, confocal laser, near infrared spectroscopy, nuclear magnetic resonance technology and ultrasound. Understanding of the mechanism of these new technologies will be helpful for applying them to improve the quality of frozen foods.

Freezing tolerance of winter wheat as influenced by extended growth at low temperature and exposure to freeze-thaw cycles

USDA-ARS?s Scientific Manuscript database

As the seasons progress, autumn-planted winter wheat plants (Triticum aestivum L.) first gain, then progressively lose freezing tolerance. Exposing the plants to freeze-thaw cycles of -3/3°C results in increased ability to tolerate subsequent freezing to potentially damaging temperatures. This stu...

Host-mediated shift in the cold tolerance of an invasive insect

Treesearch

Amy C. Morey; Robert C. Venette; Erica C. Nystrom Santacruz; Laurel A. Mosca; W. D. Hutchison

2016-01-01

While many insects cannot survive the formation of ice within their bodies, a few species can. On the evolutionary continuum from freeze-intolerant (i.e., freeze-avoidant) to freeze-tolerant insects, intermediates likely exist that can withstand some ice formation, but not enough to be considered fully freeze tolerant. Theory suggests that freeze tolerance should be...

Freezing pattern and frost killing temperature of apple (Malus domestica) wood under controlled conditions and in nature.

PubMed

Pramsohler, Manuel; Hacker, Jürgen; Neuner, Gilbert

2012-07-01

The freezing pattern and frost killing temperatures of apple (Malus domestica Borkh.) xylem were determined by differential thermal analysis and infrared differential thermal analysis (IDTA). Results from detached or attached twigs in controlled freezing experiments and during natural field freezing of trees were compared. Non-lethal freezing of apoplastic water in apple xylem as monitored during natural winter frosts in the field occurred at -1.9 ± 0.4 °C and did not change seasonally. The pattern of whole tree freezing was variable and specific to the environmental conditions. On detached twigs high-temperature freezing exotherms (HTEs) occurred 2.8 K below the temperature observed under natural frosts in the field with a seasonal mean of -4.7 ± 0.5 °C. Microporous apple xylem showed freezing without a specific pattern within a few seconds in IDTA images during HTEs, which is in contrast to macroporous xylem where a 2D freezing pattern mirrors anatomical structures. The pith tissue always remained unfrozen. Increasing twig length increased ice nucleation temperature; for increased twig diameter the effect was not significant. In attached twigs frozen in field portable freezing chambers, HTEs were recorded at a similar mean temperature (-4.6 ± 1.0 °C) to those for detached twigs. Upon lethal intracellular freezing of apple xylem parenchyma cells (XPCs) low-temperature freezing exotherms (LTEs) can be recorded. Low-temperature freezing exotherms determined on detached twigs varied significantly between a winter minimum of -36.9 °C and a summer maximum -12.7 °C. Within the temperature range wherein LTEs were recorded by IDTA in summer (-12.7 ± 0.5 to -20.3 ± 1.1 °C) various tiny clearly separated discontinuous freezing events could be detected similar to that in other species with contrasting XPC anatomy. These freezing events appeared to be initially located in the primary and only later in the secondary xylem. During the LTE no freezing events in the bark and central pith tissue were recorded. Attached twigs were exposed to various freezing temperatures at which LTEs occur. Even if 60% of XPCs were frost-damaged twigs were able to recuperate and showed full re-growth indicating a high regeneration capacity even after severe frost damage to XPCs.

Defensive freezing links Hypothalamic-Pituitary-Adrenal-axis activity and internalizing symptoms in humans.

PubMed

Niermann, Hannah C M; Figner, Bernd; Tyborowska, Anna; van Peer, Jacobien M; Cillessen, Antonius H N; Roelofs, Karin

2017-08-01

The Hypothalamic-Pituitary-Adrenal (HPA)-axis plays an important role in the expression of defensive freezing. Adaptive freezing reactivity, characterized by an immediate increase in acute stress and timely termination upon threat offset or need to act, is essential for adequate stress coping. Blunted HPA-axis activity in animals is associated with blunted freezing reactivity and internalizing symptoms. Despite their potential relevance, it remains unknown whether these mechanisms apply to humans and human psychopathology. Using a well-established method combining electrocardiography and posturography, we assessed freezing before, immediately after, and one hour after a stress induction in 92 human adolescents. In line with animal models, human adolescents showed stress-induced freezing, as quantified by relative reductions in heart rate and body sway after, as compared to before, stress. Moreover, relatively lower basal cortisol was associated with reduced stress-induced freezing reactivity (i.e., less immediate freezing and less recovery). Path analyses showed that decreased freezing recovery in individuals with reduced cortisol levels was associated with increased levels of internalizing symptoms. These findings suggest that reduced freezing recovery may be a promising marker for the etiology of internalizing symptoms. Copyright © 2017 Elsevier Ltd. All rights reserved.

Improving survival and storage stability of bacteria recalcitrant to freeze-drying: a coordinated study by European culture collections.

PubMed

Peiren, Jindrich; Buyse, Joke; De Vos, Paul; Lang, Elke; Clermont, Dominique; Hamon, Sylviane; Bégaud, Evelyne; Bizet, Chantal; Pascual, Javier; Ruvira, María A; Macián, M Carmen; Arahal, David R

2015-04-01

The objective of this study is to improve the viability after freeze-drying and during storage of delicate or recalcitrant strains safeguarded at biological resource centers. To achieve this objective, a joint experimental strategy was established among the different involved partner collections of the EMbaRC project ( www.embarc.eu ). Five bacterial strains considered as recalcitrant to freeze-drying were subjected to a standardized freeze-drying protocol and to seven agreed protocol variants. Viability of these strains was determined before and after freeze-drying (within 1 week, after 6 and 12 months, and after accelerated storage) for each of the protocols. Furthermore, strains were exchanged between partners to perform experiments with different freeze-dryer-dependent parameters. Of all tested variables, choice of the lyoprotectant had the biggest impact on viability after freeze-drying and during storage. For nearly all tested strains, skim milk as lyoprotectant resulted in lowest viability after freeze-drying and storage. On the other hand, best freeze-drying and storage conditions were strain and device dependent. For Aeromonas salmonicida CECT 894(T), best survival was obtained when horse serum supplemented with trehalose was used as lyoprotectant, while Aliivibrio fischeri LMG 4414(T) should be freeze-dried in skim milk supplemented with marine broth in a 1:1 ratio. Freeze-drying Campylobacter fetus CIP 53.96(T) using skim milk supplemented with trehalose as lyoprotectant resulted in best recovery. Xanthomonas fragariae DSM 3587(T) expressed high viability after freeze-drying and storage for all tested lyoprotectants and could not be considered as recalcitrant. In contrary, Flavobacterium columnare LMG 10406(T) did not survive the freeze-drying process under all tested conditions.

State transitions and physicochemical aspects of cryoprotection and stabilization in freeze-drying of Lactobacillus rhamnosus GG (LGG).

PubMed

Pehkonen, K S; Roos, Y H; Miao, S; Ross, R P; Stanton, C

2008-06-01

The frozen and dehydrated state transitions of lactose and trehalose were determined and studied as factors affecting the stability of probiotic bacteria to understand physicochemical aspects of protection against freezing and dehydration of probiotic cultures. Lactobacillus rhamnosus GG was frozen (-22 or -43 degrees C), freeze-dried and stored under controlled water vapour pressure (0%, 11%, 23% and 33% relative vapour pressure) conditions. Lactose, trehalose and their mixture (1 : 1) were used as protective media. These systems were confirmed to exhibit relatively similar state transition and water plasticization behaviour in freeze-concentrated and dehydrated states as determined by differential scanning calorimetry. Ice formation and dehydrated materials were studied using cold-stage microscopy and scanning electron microscopy. Trehalose and lactose-trehalose gave the most effective protection of cell viability as observed from colony forming units after freezing, dehydration and storage. Enhanced cell viability was observed when the freezing temperature was -43 degrees C. State transitions of protective media affect ice formation and cell viability in freeze-drying and storage. Formation of a maximally freeze-concentrated matrix with entrapped microbial cells is essential in freezing prior to freeze-drying. Freeze-drying must retain a solid amorphous state of protectant matrices. Freeze-dried matrices contain cells entrapped in the protective matrices in the freezing process. The retention of viability during storage seems to be controlled by water plasticization of the protectant matrix and possibly interactions of water with the dehydrated cells. Highest cell viability was obtained in glassy protective media. This study shows that physicochemical properties of protective media affect the stability of dehydrated cultures. Trehalose and lactose may be used in combination, which is particularly important for the stabilization of probiotic bacteria in dairy systems.

Experimental analysis and modeling of ultrasound assisted freezing of potato spheres.

PubMed

Kiani, Hossein; Zhang, Zhihang; Sun, Da-Wen

2015-09-01

In recent years, innovative methods such as ultrasound assisted freezing have been developed in order to improve the freezing process. During freezing of foods, accurate prediction of the temperature distribution, phase ratios, and process time is very important. In the present study, ultrasound assisted immersion freezing process (in 1:1 ethylene glycol-water solution at 253.15K) of potato spheres (0.02 m diameter) was evaluated using experimental, numerical and analytical approaches. Ultrasound (25 kHz, 890 W m(-2)) was irradiated for different duty cycles (DCs=0-100%). A finite volume based enthalpy method was used in the numerical model, based on which temperature and liquid fraction profiles were simulated by a program developed using OpenFOAM® CFD software. An analytical technique was also employed to calculate freezing times. The results showed that ultrasound irradiation could decrease the characteristic freezing time of potatoes. Since ultrasound irradiation increased the heat transfer coefficient but simultaneously generated heat at the surface of the samples, an optimum DC was needed for the shortest freezing time which occurred in the range of 30-70% DC. DCs higher than 70% increased the freezing time. DCs lower than 30% did not provide significant effects on the freezing time compared to the control sample. The numerical model predicted the characteristic freezing time in accordance with the experimental results. In addition, analytical calculation of characteristic freezing time exhibited qualitative agreement with the experimental results. As the numerical simulations provided profiles of temperature and water fraction within potatoes frozen with or without ultrasound, the models can be used to study and control different operation situations, and to improve the understanding of the freezing process. Copyright © 2015 Elsevier B.V. All rights reserved.

Numerical simulation of the effect of groundwater salinity on artificial freezing wall in coastal area

NASA Astrophysics Data System (ADS)

Hu, Rui; Liu, Quan

2017-04-01

During the engineering projects with artificial ground freezing (AFG) techniques in coastal area, the freezing effect is affected by groundwater salinity. Based on the theories of artificially frozen soil and heat transfer in porous material, and with the assumption that only the variations of total dissolved solids (TDS) impact on freezing point and thermal conductivity, a numerical model of an AFG project in a saline aquifer was established and validated by comparing the simulated temperature field with the calculated temperature based on the analytic solution of rupak (reference) for single-pipe freezing temperature field T. The formation and development of freezing wall were simulated with various TDS. The results showed that the variety of TDS caused the larger temperature difference near the frozen front. With increasing TDS in the saline aquifer (1 35g/L), the average thickness of freezing wall decreased linearly and the total formation time of the freezing wall increased linearly. Compared with of the scenario of fresh-water (<1g/L), the average thickness of frozen wall decreased by 6% and the total formation time of the freezing wall increased by 8% with each increasing TDS of 7g/L. Key words: total dissolved solids, freezing point, thermal conductivity, freezing wall, numerical simulation Reference D.J.Pringel, H.Eicken, H.J.Trodahl, etc. Thermal conductivity of landfast Antarctic and Arctic sea ice[J]. Journal of Geophysical Research, 2007, 112: 1-13. Lukas U.Arenson, Dave C.Sego. The effect of salinity on the freezing of coarse- grained sand[J]. Canadian Geotechnical Journal, 2006, 43: 325-337. Hui Bing, Wei Ma. Laboratory investigation of the freezing point of saline soil[J]. Cold Regions Science and Technology, 2011, 67: 79-88.

Mechanisms of deterioration of nutrients phase 1

NASA Technical Reports Server (NTRS)

Karel, M.; Flink, J. M.

1972-01-01

Experimental methods are studied by which freeze-dried foods of improved quality are produced. Considered are: (1) Factors effecting the loss of butanol from frozen aqueous food solutions during storage; (2) a freeze-drying microscope system for observing solidification processes in organic mixtures and aqueous inorganic salt solutions; (3) browning of high quality freeze-dried foods with minimal organoleptic and nutritional detoriation; (4) retention of PVP-n-propanol in freeze-dried food models; and (5) effects of freezing rate and sucrose immersion on taste and texture of freeze-dried apple slices.

Effect of moisture content on the invertase activity of freeze-dried S. cerevisiae.

PubMed

Pitombo, R N; Spring, C; Passos, R F; Tonato, M; Vitolo, M

1994-08-01

The invertase activity of intact Saccharomyces cerevisiae submitted to freezing-thawing was affected by pH, the chemical nature of the buffer, and the freezing cooling rate (CR), leading in some cases to a complete invertase inactivation (acetate buffer, pH 4.0, CR = 0.5 degree C/min). Once established under adequate freezing conditions the invertase activity remained unchanged after freeze-drying. Nevertheless, in some cases the cell-growing capability after freeze-drying diminished around 70%, mainly if the frozen cell suspension was attained through freezing carried out at CR = 0.5 degree C/min. Water sorption isotherms of freeze-dried samples (freeze-dryer Edwards L-4KR; 30 degrees C and 0.1 mB) were determined at 10 and 25 degrees C. The monolayer moisture content (MMC) at each temperature (12.7 and 3.71 for 10 and 25 degrees C, respectively) was calculated from isotherms by applying BET and GAB models. Freeze-dried yeast with water activity (Aw) between 0 and 0.33 (about the MMC value) maintained at 25 degrees C for 235 days and at 89 degrees C for 15 min retained at least 85% of its original invertase activity (IA), whereas samples with Aw > MMC lost at least 60% of its IA. X ray diffraction showed that the freeze-dried cake before and after storage presented an amorphous structure.

Ice-Binding Proteins in Plants

PubMed Central

Bredow, Melissa; Walker, Virginia K.

2017-01-01

Sub-zero temperatures put plants at risk of damage associated with the formation of ice crystals in the apoplast. Some freeze-tolerant plants mitigate this risk by expressing ice-binding proteins (IBPs), that adsorb to ice crystals and modify their growth. IBPs are found across several biological kingdoms, with their ice-binding activity and function uniquely suited to the lifestyle they have evolved to protect, be it in fishes, insects or plants. While IBPs from freeze-avoidant species significantly depress the freezing point, plant IBPs typically have a reduced ability to lower the freezing temperature. Nevertheless, they have a superior ability to inhibit the recrystallization of formed ice. This latter activity prevents ice crystals from growing larger at temperatures close to melting. Attempts to engineer frost-hardy plants by the controlled transfer of IBPs from freeze-avoiding fish and insects have been largely unsuccessful. In contrast, the expression of recombinant IBP sequences from freeze-tolerant plants significantly reduced electrolyte leakage and enhanced freezing survival in freeze-sensitive plants. These promising results have spurred additional investigations into plant IBP localization and post-translational modifications, as well as a re-evaluation of IBPs as part of the anti-stress and anti-pathogen axis of freeze-tolerant plants. Here we present an overview of plant freezing stress and adaptation mechanisms and discuss the potential utility of IBPs for the generation of freeze-tolerant crops. PMID:29312400

Ice-Binding Proteins in Plants.

PubMed

Bredow, Melissa; Walker, Virginia K

2017-01-01

Sub-zero temperatures put plants at risk of damage associated with the formation of ice crystals in the apoplast. Some freeze-tolerant plants mitigate this risk by expressing ice-binding proteins (IBPs), that adsorb to ice crystals and modify their growth. IBPs are found across several biological kingdoms, with their ice-binding activity and function uniquely suited to the lifestyle they have evolved to protect, be it in fishes, insects or plants. While IBPs from freeze-avoidant species significantly depress the freezing point, plant IBPs typically have a reduced ability to lower the freezing temperature. Nevertheless, they have a superior ability to inhibit the recrystallization of formed ice. This latter activity prevents ice crystals from growing larger at temperatures close to melting. Attempts to engineer frost-hardy plants by the controlled transfer of IBPs from freeze-avoiding fish and insects have been largely unsuccessful. In contrast, the expression of recombinant IBP sequences from freeze-tolerant plants significantly reduced electrolyte leakage and enhanced freezing survival in freeze-sensitive plants. These promising results have spurred additional investigations into plant IBP localization and post-translational modifications, as well as a re-evaluation of IBPs as part of the anti-stress and anti-pathogen axis of freeze-tolerant plants. Here we present an overview of plant freezing stress and adaptation mechanisms and discuss the potential utility of IBPs for the generation of freeze-tolerant crops.

Changes in Ultrastructure and Sensory Characteristics on Electro-magnetic and Air Blast Freezing of Beef during Frozen Storage

PubMed Central

2015-01-01

The ultrastructure in the beef muscle of the electro-magnetic resonance and air blast freezing during the frozen storage, and the changes in the quality characteristics after thawing were evaluated. The size of ice crystal was small and evenly formed in the initial freezing period, and it showed that the size was increased as the storage period was elapsed (p<0.05). The beef stored by the electro-magnetic resonance freezing showed the size of ice crystal with a lower rate of increase than the air blast freezing during the frozen storage. The thawing loss of beef stored by the electro-magnetic resonance freezing was significantly lower than the air blast freezing during frozen storage (p<0.05), and it showed that the thawing loss of the round was higher than the loin. Water holding capacity decreased as the storage period became longer while the electro-magnetic resonance freezing was higher than the air blast on 8 month (p<0.05). As a result of sensory evaluation, the beef stored by the electro-magnetic resonance freezing did not show the difference until 4 months, and it showed higher acceptability in comparison with the beef stored by the air blast freezing. Thus, it is considered that the freezing method has an effect on the change in the ultrastructure and quality characteristics of the beef. PMID:26761797

Freeze-thaw durability of concrete: Ice formation process in pores

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cai, H.; Liu, X.

1998-09-01

Freeze-thaw durability of concrete is of great importance to hydraulic structures in cold areas. Study of ice formation process in concrete pores is necessary to evaluate the damages in concrete caused by freezing. In this paper, freezing of pore solution in concrete exposed to a freeze-thaw cycle is studied by following the change of concrete electrical conductivity with freezing temperatures. Concretes were subjected to freeze-thaw cycles with temperature varying between {minus}0 C and {minus}20 C. In the freezing process, the changing rate of concrete electrical conductivity obviously decreases at about {minus}10 C, indicating that more pore solution in concrete freezesmore » above {minus}10 C than below {minus}10C. According to Powers` static hydraulic pressure hypothesis, it is thought that frost damage mainly occurs between 0 C and {minus}100 C. To ordinary concrete, frost damages below {minus}10 C are negligible.« less

Fish antifreeze protein and the freezing and recrystallization of ice.

PubMed

Knight, C A; DeVries, A L; Oolman, L D

Antifreeze glycopeptide and peptides from the blood of polar fishes prevent the growth of ice crystals in water at temperatures down to approximately 1 degree C below freezing point, but do not appreciably influence the equilibrium freezing point. This freezing point hysteresis must be a disequilibrium effect, or it would violate Gibbs' phase rule, but the separate freezing and melting points are experimentally very definite: ice neither melts nor freezes perceptibly within the 'hysteresis gap', for periods of hours or days. We report here unusual crystal faces on ice crystals grown from solutions of very low concentrations of the anti-freeze glycopeptides and peptides. This is a clue to the mechanism of freezing inhibition, and it may be the basis of a simple, very sensitive test for antifreeze material. Very low concentrations of the antifreeze protein are also remarkably effective in preventing the recrystallization of ice.

Increased spring freezing vulnerability for alpine shrubs under early snowmelt.

PubMed

Wheeler, J A; Hoch, G; Cortés, A J; Sedlacek, J; Wipf, S; Rixen, C

2014-05-01

Alpine dwarf shrub communities are phenologically linked with snowmelt timing, so early spring exposure may increase risk of freezing damage during early development, and consequently reduce seasonal growth. We examined whether environmental factors (duration of snow cover, elevation) influenced size and the vulnerability of shrubs to spring freezing along elevational gradients and snow microhabitats by modelling the past frequency of spring freezing events. We sampled biomass and measured the size of Salix herbacea, Vaccinium myrtillus, Vaccinium uliginosum and Loiseleuria procumbens in late spring. Leaves were exposed to freezing temperatures to determine the temperature at which 50% of specimens are killed for each species and sampling site. By linking site snowmelt and temperatures to long-term climate measurements, we extrapolated the frequency of spring freezing events at each elevation, snow microhabitat and per species over 37 years. Snowmelt timing was significantly driven by microhabitat effects, but was independent of elevation. Shrub growth was neither enhanced nor reduced by earlier snowmelt, but decreased with elevation. Freezing resistance was strongly species dependent, and did not differ along the elevation or snowmelt gradient. Microclimate extrapolation suggested that potentially lethal freezing events (in May and June) occurred for three of the four species examined. Freezing events never occurred on late snow beds, and increased in frequency with earlier snowmelt and higher elevation. Extrapolated freezing events showed a slight, non-significant increase over the 37-year record. We suggest that earlier snowmelt does not enhance growth in four dominant alpine shrubs, but increases the risk of lethal spring freezing exposure for less freezing-resistant species.

Controlled crystallization of the lipophilic drug fenofibrate during freeze-drying: elucidation of the mechanism by in-line Raman spectroscopy.

PubMed

de Waard, Hans; De Beer, Thomas; Hinrichs, Wouter L J; Vervaet, Chris; Remon, Jean-Paul; Frijlink, Henderik W

2010-12-01

We developed a novel process, "controlled crystallization during freeze-drying" to produce drug nanocrystals of poorly water-soluble drugs. This process involves freeze-drying at a relatively high temperature of a drug and a matrix material from a mixture of tertiary butyl alcohol and water, resulting in drug nanocrystals incorporated in a matrix. The aim of this study was to elucidate the mechanisms that determine the size of the drug crystals. Fenofibrate was used as a model lipophilic drug. To monitor the crystallization during freeze-drying, a Raman probe was placed just above the sample in the freeze-dryer. These in-line Raman spectroscopy measurements clearly revealed when the different components crystallized during freeze-drying. The solvents crystallized only during the freezing step, while the solutes only crystallized after the temperature was increased, but before drying started. Although the solutes crystallized only after the freezing step, both the freezing rate and the shelf temperature were critical parameters that determined the final crystal size. At a higher freezing rate, smaller interstitial spaces containing the freeze-concentrated fraction were formed, resulting in smaller drug crystals (based on dissolution data). On the other hand, when the solutes crystallized at a lower shelf temperature, the degree of supersaturation is higher, resulting in a higher nucleation rate and consequently more and therefore smaller crystals. In conclusion, for the model drug fenofibrate, a high freezing rate and a relatively low crystallization temperature resulted in the smallest crystals and therefore the highest dissolution rate.

Effect of three cueing devices for people with Parkinson's disease with gait initiation difficulties.

PubMed

McCandless, Paula J; Evans, Brenda J; Janssen, Jessie; Selfe, James; Churchill, Andrew; Richards, Jim

2016-02-01

Freezing of gait (FOG) remains one of the most common debilitating aspects of Parkinson's disease and has been linked to injuries, falls and reduced quality of life. Although commercially available portable cueing devices exist claiming to assist with overcoming freezing; their immediate effectiveness in overcoming gait initiation failure is currently unknown. This study investigated the effects of three different types of cueing device in people with Parkinson's disease who experience freezing. Twenty participants with idiopathic Parkinson's disease who experienced freezing during gait but who were able to walk short distances indoors independently were recruited. At least three attempts at gait initiation were recorded using a 10 camera Qualisys motion analysis system and four force platforms. Test conditions were; Laser Cane, sound metronome, vibrating metronome, walking stick and no intervention. During testing 12 of the 20 participants had freezing episodes, from these participants 100 freezing and 91 non-freezing trials were recorded. Clear differences in the movement patterns were seen between freezing and non-freezing episodes. The Laser Cane was most effective cueing device at improving the forwards/backwards and side to side movement and had the least number of freezing episodes. The walking stick also showed significant improvements compared to the other conditions. The vibration metronome appeared to disrupt movement compared to the sound metronome at the same beat frequency. This study identified differences in the movement patterns between freezing episodes and non-freezing episodes, and identified immediate improvements during gait initiation when using the Laser Cane over the other interventions. Copyright © 2015. Published by Elsevier B.V.

Role of glucose-6-phosphate dehydrogenase in freezing-induced freezing resistance of Populus suaveolens.

PubMed

Lin, Shan-Zhi; Zhang, Zhi-Yi; Liu, Wen-Feng; Lin, Yuan-Zhen; Zhang, Qian; Zhu, Bao-Qing

2005-02-01

To explore the role of glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) in the enhancement of freezing resistance induced by freezing acclimation, G6PDH was purified from the leaves of 8-week-old Populus suaveolens cuttings. The G6PDH activity in the absence or the presence of reduced dithiothreitol (DTT(red)) were determined, and the changes in superoxide dismutase (SOD), peroxides (POD) and cytosolic G6PDH activities, malondial-dehyde (MDA) content as well as freezing resistance (expressed as LT(50)) of P. suaveolens cuttings during freezing acclimation at -20 degrees C were investigated. The results showed that the purified G6PDH was probably located in the cytosol of P. suaveolens. Freezing acclimation increased the activities of SOD, POD and cytosolic G6PDH, and decreased the MDA content and LT(50) of cuttings, while 2 d of de-acclimation at 25 degrees C resulted in a decrease in SOD, POD and cytosolic G6PDH activities, and caused an increase in MDA content and LT(50). The change in cytosolic G6PDH activity was found to be closely correlated to the levels of SOD, POD and MDA, and to the degree of freezing resistance of cuttings during freezing acclimation. It is suggested that the enhancement of freezing resistance of cuttings induced by freezing acclimation is related to the distinct increase in cytosolic G6PDH activity, which may be involved in the activation of SOD and POD, and the induction of freezing resistance of cuttings.

Differential freezing resistance and photoprotection in C3 and C4 eudicots and grasses

PubMed Central

Liu, Mei-Zhen; Osborne, Colin P.

2013-01-01

Globally, C4 plants dominate hot, open environments, but this general pattern is underpinned by important differences in the biogeography of C4 lineages. In particular, the species richness of C4 Poaceae (grasses) increases strongly with increasing temperature, whereas that of the major C4 eudicot group Chenopodiaceae correlates positively with aridity. Freezing tolerance is a crucial determinant of biogeographical relationships with temperature and is mediated by photodamage and cellular disruption by desiccation, but little is known about differences between C4 families. This study hypothesized that there is a greater risk of freezing damage via these mechanisms in C4 Poaceae than Chenopodiaceae, that freezing protection differs between the taxonomic groups, and that freezing tolerance of species is linked to arid habitat preference. Chlorophyll fluorescence, water relations, and freezing injury were compared in four C3 and six C4 species of Poaceae and Chenopodiaceae from the same Mongolian flora. Contrary to expectations, freezing-induced leaf mortality and photodamage were lower in Poaceae than Chenopodiaceae species, and unrelated to photosynthetic pathway. The freezing resistance of Poaceae species resulted from constitutive protection and cold acclimation and an ability to protect the photosynthetic apparatus from photodamage. Freezing protection was associated with low osmotic potential and low tissue elasticity, and freezing damage was accompanied by electrolyte leakage, consistent with cell-membrane disruption by ice. Both Chenopodiaceae and Poaceae had the potential to develop cold acclimation and withstand freezing during the growing season, which conflicted with the hypothesis. Instead, freezing tolerance was more closely associated with life history and ecological preference in these Mongolian species. PMID:23599273

Embolism Formation during Freezing in the Wood of Picea abies1

PubMed Central

Mayr, Stefan; Cochard, Hervé; Améglio, Thierry; Kikuta, Silvia B.

2007-01-01

Freeze-thaw events can cause embolism in plant xylem. According to classical theory, gas bubbles are formed during freezing and expand during thawing. Conifers have proved to be very resistant to freeze-thaw induced embolism, because bubbles in tracheids are small and redissolve during thawing. In contrast, increasing embolism rates upon consecutive freeze-thaw events were observed that cannot be explained by the classical mechanism. In this study, embolism formation during freeze-thaw events was analyzed via ultrasonic and Cryo-scanning electron microscope techniques. Twigs of Picea abies L. Karst. were subjected to up to 120 freeze-thaw cycles during which ultrasonic acoustic emissions, xylem temperature, and diameter variations were registered. In addition, the extent and cross-sectional pattern of embolism were analyzed with staining experiments and Cryo-scanning electron microscope observations. Embolism increased with the number of freeze-thaw events in twigs previously dehydrated to a water potential of −2.8 MPa. In these twigs, acoustic emissions were registered, while saturated twigs showed low, and totally dehydrated twigs showed no, acoustic activity. Acoustic emissions were detected only during the freezing process. This means that embolism was formed during freezing, which is in contradiction to the classical theory of freeze-thaw induced embolism. The clustered pattern of embolized tracheids in cross sections indicates that air spread from a dysfunctional tracheid to adjacent functional ones. We hypothesize that the low water potential of the growing ice front led to a decrease of the potential in nearby tracheids. This may result in freezing-induced air seeding. PMID:17041033

Contact freezing of supercooled cloud droplets on collision with mineral dust particles: effect of particle size

NASA Astrophysics Data System (ADS)

Hoffmann, Nadine; Duft, Denis; Kiselev, Alexei; Leisner, Thomas

2013-04-01

The contact freezing of supercooled cloud droplets is one of the potentially important and the least investigated heterogeneous mechanism of ice formation in the tropospheric clouds [1]. On the time scales of cloud lifetime the freezing of supercooled water droplets via contact mechanism may occur at higher temperature compared to the same IN immersed in the droplet. However, the laboratory experiments of contact freezing are very challenging due to the number of factors affecting the probability of ice formation. In our experiment we study single water droplets freely levitated in the laminar flow of mineral dust particles acting as the contact freezing nuclei. By repeating the freezing experiment sufficient number of times we are able to reproduce statistical freezing behavior of large ensembles of supercooled droplets and measure the average rate of freezing events. We show that the rate of freezing at given temperature is governed only by the rate of droplet -particle collision and by the properties of the contact ice nuclei. In this contribution we investigate the relationship between the freezing probability and the size of mineral dust particle (represented by illite) and show that their IN efficiency scales with the particle size. Based on this observation, we discuss the similarity between the freezing of supercooled water droplets in immersion and contact modes and possible mechanisms of apparent enhancement of the contact freezing efficiency. [1] - K.C. Young, The role of contact nucleation in ice phase initiation in clouds, Journal of the Atmospheric Sciences 31, 1974

Ice nucleation temperature influences recovery of activity of a model protein after freeze drying.

PubMed

Cochran, Teresa; Nail, Steven L

2009-09-01

The objective of this study was to determine whether a relationship exists between ice nucleation temperature and recovery of activity of a model protein, lactate dehydrogenase, after freeze drying. Aqueous buffer systems containing 50 microg/mL of protein were frozen in vials with externally mounted thermocouples on the shelf of a freeze dryer, then freeze dried. Various methods were used to establish a wide range of ice nucleation temperatures. An inverse relationship was found between the extent of supercooling during freezing and recovery of activity in the reconstituted solution. The data are consistent with a mechanism of inactivation resulting from adsorption of protein at the ice/freeze-concentrate interface during the freezing process.

Predictive modeling of freezing and thawing of frost-susceptible soils.

DOT National Transportation Integrated Search

2015-09-01

Frost depth is an essential factor in design of various transportation infrastructures. In frost : susceptible soils, as soils freezes, water migrates through the soil voids below the freezing line : towards the freezing front and causes excessive he...

Cognitive Contributions to Freezing of Gait in Parkinson Disease: Implications for Physical Rehabilitation

PubMed Central

King, Laurie A.; Cohen, Rajal G.; Horak, Fay B.

2016-01-01

People with Parkinson disease (PD) who show freezing of gait also have dysfunction in cognitive domains that interact with mobility. Specifically, freezing of gait is associated with executive dysfunction involving response inhibition, divided attention or switching attention, and visuospatial function. The neural control impairments leading to freezing of gait have recently been attributed to higher-level, executive and attentional cortical processes involved in coordinating posture and gait rather than to lower-level, sensorimotor impairments. To date, rehabilitation for freezing of gait primarily has focused on compensatory mobility training to overcome freezing events, such as sensory cueing and voluntary step planning. Recently, a few interventions have focused on restitutive, rather than compensatory, therapy. Given the documented impairments in executive function specific to patients with PD who freeze and increasing evidence of overlap between cognitive and motor function, incorporating cognitive challenges with mobility training may have important benefits for patients with freezing of gait. Thus, a novel theoretical framework is proposed for exercise interventions that jointly address both the specific cognitive and mobility challenges of people with PD who freeze. PMID:26381808

Cognitive Contributions to Freezing of Gait in Parkinson Disease: Implications for Physical Rehabilitation.

PubMed

Peterson, Daniel S; King, Laurie A; Cohen, Rajal G; Horak, Fay B

2016-05-01

People with Parkinson disease (PD) who show freezing of gait also have dysfunction in cognitive domains that interact with mobility. Specifically, freezing of gait is associated with executive dysfunction involving response inhibition, divided attention or switching attention, and visuospatial function. The neural control impairments leading to freezing of gait have recently been attributed to higher-level, executive and attentional cortical processes involved in coordinating posture and gait rather than to lower-level, sensorimotor impairments. To date, rehabilitation for freezing of gait primarily has focused on compensatory mobility training to overcome freezing events, such as sensory cueing and voluntary step planning. Recently, a few interventions have focused on restitutive, rather than compensatory, therapy. Given the documented impairments in executive function specific to patients with PD who freeze and increasing evidence of overlap between cognitive and motor function, incorporating cognitive challenges with mobility training may have important benefits for patients with freezing of gait. Thus, a novel theoretical framework is proposed for exercise interventions that jointly address both the specific cognitive and mobility challenges of people with PD who freeze. © 2016 American Physical Therapy Association.

Representative Values of Icing-Related Variables Aloft in Freezing Rain and Freezing Drizzle

DOT National Transportation Integrated Search

1996-03-01

Radiosonde and surface observations in freezing rain (ZR) and freezing drizzle (ZL), and a limited number of aircraft measurements in ZR, have been examined for information on the magnitude and altitude dependence of meteorological variables associat...

Effects of Artificial Supercooling Followed by Slow Freezing on the Microstructure and Qualities of Pork Loin

PubMed Central

2016-01-01

This study investigated the effects of artificial supercooling followed by still air freezing (SSF) on the qualities of pork loin. The qualities of pork frozen by SSF were compared with the fresh control (CT, stored at 4℃ for 24 h), slow freezing (SAF, still air freezing) and rapid freezing (EIF, ethanol immersion freezing) treatments. Compared with no supercooling phenomena of SAF and EIF, the extent of supercooling obtained by SSF treatment was 1.4℃. Despite that SSF was conducted with the same method with SAF, application of artificial supercooling accelerated the phase transition (traverse from -0.6℃ to -5℃) from 3.07 h (SAF) to 2.23 h (SSF). The observation of a microstructure indicated that the SSF prevented tissue damage caused by ice crystallization and maintained the structural integrity. The estimated quality parameters reflected that SSF exhibited superior meat quality compared with slow freezing (SAF). SSF showed better water-holding capacity (lower thawing loss, cooking loss and expressible moisture) and tenderness than SAF, and these quality parameters of SSF were not significantly different with ultra-fast freezing treatment (EIF). Consequently, the results demonstrated that the generation of supercooling followed by conventional freezing potentially had the advantage of minimizing the quality deterioration caused by the slow freezing of meat. PMID:27857541

Demonstrating Functional Equivalence of Pilot and Production Scale Freeze-Drying of BCG

PubMed Central

ten Have, R.; Reubsaet, K.; van Herpen, P.; Kersten, G.; Amorij, J.-P.

2016-01-01

Process analytical technology (PAT)-tools were used to monitor freeze-drying of Bacille Calmette-Guérin (BCG) at pilot and production scale. Among the evaluated PAT-tools, there is the novel use of the vacuum valve open/close frequency for determining the endpoint of primary drying at production scale. The duration of primary drying, the BCG survival rate, and the residual moisture content (RMC) were evaluated using two different freeze-drying protocols and were found to be independent of the freeze-dryer scale evidencing functional equivalence. The absence of an effect of the freeze-dryer scale on the process underlines the feasibility of the pilot scale freeze-dryer for further BCG freeze-drying process optimization which may be carried out using a medium without BCG. PMID:26981867

Demonstrating Functional Equivalence of Pilot and Production Scale Freeze-Drying of BCG.

PubMed

Ten Have, R; Reubsaet, K; van Herpen, P; Kersten, G; Amorij, J-P

2016-01-01

Process analytical technology (PAT)-tools were used to monitor freeze-drying of Bacille Calmette-Guérin (BCG) at pilot and production scale. Among the evaluated PAT-tools, there is the novel use of the vacuum valve open/close frequency for determining the endpoint of primary drying at production scale. The duration of primary drying, the BCG survival rate, and the residual moisture content (RMC) were evaluated using two different freeze-drying protocols and were found to be independent of the freeze-dryer scale evidencing functional equivalence. The absence of an effect of the freeze-dryer scale on the process underlines the feasibility of the pilot scale freeze-dryer for further BCG freeze-drying process optimization which may be carried out using a medium without BCG.

Research on strength attenuation law of concrete in freezing - thawing environment

NASA Astrophysics Data System (ADS)

Xiao, qianhui; Cao, zhiyuan; Li, qiang

2018-03-01

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.

Experimental research on durability of recycled aggregate concrete under freeze- thaw cycles

NASA Astrophysics Data System (ADS)

Cheng, Yanqiu; Shang, Xiaoyu; Zhang, Youjia

2017-07-01

The freeze-thaw durability of recycled aggregate concrete has significance for the concrete buildings in the cold region. In this paper, the rapid freezing and thawing cycles experience on recycle aggregate concrete was conducted to study on the effects of recycle aggregate amount, water-binder ratio and fly ash on freeze-thaw durability of recycle aggregate concrete. The results indicates that recycle aggregate amount makes the significant influence on the freeze-thaw durability. With the increase of recycled aggregates amount, the freeze-thaw resistance for recycled aggregate concrete decreases. Recycled aggregate concrete with lower water cement ratio demonstrates better performance of freeze-thaw durability. It is advised that the amount of fly ash is less than 30% for admixture of recycled aggregates in the cold region.

MAL62 overexpression and NTH1 deletion enhance the freezing tolerance and fermentation capacity of the baker's yeast in lean dough.

PubMed

Sun, Xi; Zhang, Cui-Ying; Wu, Ming-Yue; Fan, Zhi-Hua; Liu, Shan-Na; Zhu, Wen-Bi; Xiao, Dong-Guang

2016-04-04

Trehalose is related to several types of stress responses, especially freezing response in baker's yeast (Saccharomyces cerevisiae). It is desirable to manipulate trehalose-related genes to create yeast strains that better tolerate freezing-thaw stress with improved fermentation capacity, which are in high demand in the baking industry. The strain overexpressing MAL62 gene showed increased trehalose content and cell viability after prefermention-freezing and long-term frozen. Deletion of NTH1 in combination of MAL62 overexpression further strengthens freezing tolerance and improves the leavening ability after freezing-thaw stress. The mutants of the industrial baker's yeast with enhanced freezing tolerance and leavening ability in lean dough were developed by genetic engineering. These strains had excellent potential industrial applications.

Freeze/Thaw-Induced Embolism: Probability of Critical Bubble Formation Depends on Speed of Ice Formation

DOE PAGES

Sevanto, Sanna; Holbrook, N. Michele; Ball, Marilyn C.

2012-06-06

Bubble formation in the conduits of woody plants sets a challenge for uninterrupted water transportation from the soil up to the canopy. Freezing and thawing of stems has been shown to increase the number of air-filled (embolized) conduits, especially in trees with large conduit diameters. Despite numerous experimental studies, the mechanisms leading to bubble formation during freezing have not been addressed theoretically. We used classical nucleation theory and fluid mechanics to show which mechanisms are most likely to be responsible for bubble formation during freezing and what parameters determine the likelihood of the process. Our results confirm the common assumptionmore » that bubble formation during freezing is most likely due to gas segregation by ice. If xylem conduit walls are not permeable to the salts expelled by ice during the freezing process, osmotic pressures high enough for air seeding could be created. The build-up rate of segregated solutes in front of the ice-water interface depends equally on conduit diameter and freezing velocity. Therefore, bubble formation probability depends on these variables. The dependence of bubble formation probability on freezing velocity means that the experimental results obtained for cavitation threshold conduit diameters during freeze/thaw cycles depend on the experimental setup; namely sample size and cooling rate. The velocity dependence also suggests that to avoid bubble formation during freezing trees should have narrow conduits where freezing is likely to be fast (e.g., branches or outermost layer of the xylem). Avoidance of bubble formation during freezing could thus be one piece of the explanation why xylem conduit size of temperate and boreal zone trees varies quite systematically.« less

Optimizing the Protocol for Pulmonary Cryoablation: A Comparison of a Dual- and Triple-Freeze Protocol

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hinshaw, J. Louis, E-mail: jhinshaw@uwhealth.or; Littrup, Peter J.; Durick, Nathan

2010-12-15

The purpose of this study was to compare a double freeze-thaw protocol to a triple freeze-thaw protocol for pulmonary cryoablation utilizing an in vivo porcine lung model. A total of 18 cryoablations were performed in normal porcine lung utilizing percutaneous technique with 9 each in a double- (10-5-10) and triple-freeze (3-3-7-7-5) protocol. Serial noncontrast CT images were obtained during the ablation. CT imaging findings and pathology were reviewed. No imaging changes were identified during the initial freeze cycle with either protocol. However, during the first thaw cycle, a region of ground glass opacity developed around the probe with both protocols.more » Because the initial freeze was shorter with the triple freeze-thaw protocol, the imaging findings were apparent sooner with this protocol (6 vs. 13 min). Also, despite a shorter total freeze time (15 vs. 20 min), the ablation zone identified with the triple freeze-thaw protocol was not significantly different from the double freeze-thaw protocol (mean diameter: 1.67 {+-} 0.41 cm vs. 1.66 {+-} 0.21 cm, P = 0.77; area: 2.1 {+-} 0.48 cm{sup 2} vs. 1.99 {+-} 0.62 cm{sup 2}, P = 0.7; and circularity: 0.95 {+-} 0.04 vs. 0.96 {+-} 0.03, P = 0.62, respectively). This study suggests that there may be several advantages of a triple freeze-thaw protocol for pulmonary cryoablation, including earlier identification of the imaging findings associated with the ablation, the promise of a shorter procedure time or larger zones of ablation, and theoretically, more effective cytotoxicity related to the additional freeze-thaw cycle.« less

Rapid freezing without cooling equilibration in canine sperm.

PubMed

Kim, Suhee; Lee, Yongcheol; Yang, Honghyun; Kim, Yong-Jun

2012-01-01

The aim of this study was to develop a rapid method of canine semen freezing without cooling equilibration using treatment with different cryoprotectant agents (CPAs) and freezing in liquid nitrogen (LN(2)) vapor in a 0.5-mL straw via modifying vitrification. Ejaculates from eight beagle dogs were frozen with different CPAs (CPA-free, 5% glycerol, 5% ethylene glycol, and 10% ethylene glycol) and freezing times (direct plunging into LN(2) or freezing for 1, 2, 3, or 10 min in LN(2) vapor before plunging into LN(2)). Frozen-thawed sperm were evaluated for motility, viability, normal morphology, and plasma- and acrosome-membrane integrities. The 5% glycerol treatment resulted in improved sperm motility, plasma-membrane integrity and acrosome-membrane integrity (P<0.05). Freezing in LN(2) vapor showed improved sperm motility, viability, and plasma membrane integrity (P<0.05), and freezing for more than 2 min in LN(2) vapor increased acrosome-membrane integrity compared with direct plunging into LN(2) (P<0.05). The direct plunging into LN(2) showed no motile sperm. However, freezing for more than 2 min in LN(2) vapor increased the total abnormalities compared to direct plunging into LN(2) (P<0.05). In conclusion, use of 5% glycerol and freezing in LN(2) vapor were essential for the rapid freezing of canine sperm without cooling equilibration. In particular, holding for 2 min in LN(2) vapor was sufficient to yield successful rapid freezing. This rapid freezing method is simple and effective in canine sperm and would be helpful to offer information for trial of vitrification in large volumes of canine sperm. Copyright © 2012 Elsevier B.V. All rights reserved.

Freeze/Thaw-induced embolism: probability of critical bubble formation depends on speed of ice formation.

PubMed

Sevanto, Sanna; Holbrook, N Michele; Ball, Marilyn C

2012-01-01

Bubble formation in the conduits of woody plants sets a challenge for uninterrupted water transportation from the soil up to the canopy. Freezing and thawing of stems has been shown to increase the number of air-filled (embolized) conduits, especially in trees with large conduit diameters. Despite numerous experimental studies, the mechanisms leading to bubble formation during freezing have not been addressed theoretically. We used classical nucleation theory and fluid mechanics to show which mechanisms are most likely to be responsible for bubble formation during freezing and what parameters determine the likelihood of the process. Our results confirm the common assumption that bubble formation during freezing is most likely due to gas segregation by ice. If xylem conduit walls are not permeable to the salts expelled by ice during the freezing process, osmotic pressures high enough for air seeding could be created. The build-up rate of segregated solutes in front of the ice-water interface depends equally on conduit diameter and freezing velocity. Therefore, bubble formation probability depends on these variables. The dependence of bubble formation probability on freezing velocity means that the experimental results obtained for cavitation threshold conduit diameters during freeze/thaw cycles depend on the experimental setup; namely sample size and cooling rate. The velocity dependence also suggests that to avoid bubble formation during freezing trees should have narrow conduits where freezing is likely to be fast (e.g., branches or outermost layer of the xylem). Avoidance of bubble formation during freezing could thus be one piece of the explanation why xylem conduit size of temperate and boreal zone trees varies quite systematically.

Mutagenic effect of freezing on mitochondrial DNA of Saccharomyces cerevisiae.

PubMed

Stoycheva, T; Venkov, P; Tsvetkov, Ts

2007-06-01

Although suggested in some studies, the mutagenic effect of freezing has not been proved by induction and isolation of mutants. Using a well-defined genetic model, we supply in this communication evidence for the mutagenic effect of freezing on mitochondrial DNA (mtDNA) of the yeast Saccharomyces cerevisiae. The cooling for 2 h at +4 degrees C, followed by freezing for 1 h at -10 degrees C and 16 h at -20 degrees C resulted in induction of respiratory mutations. The immediate freezing in liquid nitrogen was without mutagenic effect. The study of the stepwise procedure showed that the induction of respiratory mutants takes place during the freezing at -10 and -20 degrees C of cells pre-cooled at +4 degrees C. The genetic crosses of freeze-induced mutants evidenced their mitochondrial rho- origin. The freeze-induced rho- mutants are most likely free of simultaneous nuclear mutations. The extracellular presence of cryoprotectants did not prevent the mutagenic effect of freezing while accumulation of cryoprotectors inside cells completely escaped mtDNA from cryodamage. Although the results obtained favor the notion that the mutagenic effect of freezing on yeast mtDNA is due to formation and growth of intracellular ice crystals, other reasons, such as impairment of mtDNA replication or elevated levels of ROS production are discussed as possible explanations of the mutagenic effect of freezing. It is concluded that: (i) freezing can be used as a method for isolation of mitochondrial mutants in S. cerevisiae and (ii) given the substantial development in cryopreservation of cells and tissues, special precautions should be made to avoid mtDNA damage during the cryopreservation procedures.

SENSITIVE TO FREEZING2 Aids in Resilience to Salt and Drought in Freezing-Sensitive Tomato

DOE PAGES

Wang, Kun; Hersh, Hope Lynn; Benning, Christoph

2016-09-06

SENSITIVE TO FREEZING2 (SFR2) is crucial for protecting chloroplast membranes following freezing in Arabidopsis (Arabidopsis thaliana). It has been shown that SFR2 homologs are present in all land plants, including freezing-sensitive species, raising the question of SFR2 function beyond freezing tolerance. Similar to freezing, salt and drought can cause dehydration. Thus, it is hypothesized that in freezing-sensitive plants SFR2 may play roles in their resilience to salt or drought. To test this hypothesis, SlSFR2 RNAi lines were generated in the cold/freezing-sensitive species tomato (Solanum lycopersicum [M82 cv]). Hypersensitivity to salt and drought of SlSFR2-RNAi lines was observed. Higher tolerance ofmore » wild-type tomatoes was correlated with the production of trigalactosyldiacylglycerol, a product of SFR2 activity. Tomato SFR2 in vitro activity is Mg 2+-dependent and its optimal pH is 7.5, similar to that of Arabidopsis SFR2, but the specific activity of tomato SFR2 in vitro is almost double that of Arabidopsis SFR2. When salt and drought stress were applied to Arabidopsis, no conditions could be identified at which SFR2 was induced prior to irreversibly impacting plant growth, suggesting that SFR2 protects Arabidopsis primarily against freezing. Discovery of tomato SFR2 function in drought and salt resilience provides further insights into general membrane lipid remodeling-based stress tolerance mechanisms and together with protection against freezing in freezing-resistant plants such as Arabidopsis, it adds lipid remodeling as a possible target for the engineering of abiotic stress-resilient crops.« less

SENSITIVE TO FREEZING2 Aids in Resilience to Salt and Drought in Freezing-Sensitive Tomato

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Kun; Hersh, Hope Lynn; Benning, Christoph

SENSITIVE TO FREEZING2 (SFR2) is crucial for protecting chloroplast membranes following freezing in Arabidopsis (Arabidopsis thaliana). It has been shown that SFR2 homologs are present in all land plants, including freezing-sensitive species, raising the question of SFR2 function beyond freezing tolerance. Similar to freezing, salt and drought can cause dehydration. Thus, it is hypothesized that in freezing-sensitive plants SFR2 may play roles in their resilience to salt or drought. To test this hypothesis, SlSFR2 RNAi lines were generated in the cold/freezing-sensitive species tomato (Solanum lycopersicum [M82 cv]). Hypersensitivity to salt and drought of SlSFR2-RNAi lines was observed. Higher tolerance ofmore » wild-type tomatoes was correlated with the production of trigalactosyldiacylglycerol, a product of SFR2 activity. Tomato SFR2 in vitro activity is Mg 2+-dependent and its optimal pH is 7.5, similar to that of Arabidopsis SFR2, but the specific activity of tomato SFR2 in vitro is almost double that of Arabidopsis SFR2. When salt and drought stress were applied to Arabidopsis, no conditions could be identified at which SFR2 was induced prior to irreversibly impacting plant growth, suggesting that SFR2 protects Arabidopsis primarily against freezing. Discovery of tomato SFR2 function in drought and salt resilience provides further insights into general membrane lipid remodeling-based stress tolerance mechanisms and together with protection against freezing in freezing-resistant plants such as Arabidopsis, it adds lipid remodeling as a possible target for the engineering of abiotic stress-resilient crops.« less

SENSITIVE TO FREEZING2 Aids in Resilience to Salt and Drought in Freezing-Sensitive Tomato1[OPEN

PubMed Central

Hersh, Hope Lynn

2016-01-01

SENSITIVE TO FREEZING2 (SFR2) is crucial for protecting chloroplast membranes following freezing in Arabidopsis (Arabidopsis thaliana). It has been shown that SFR2 homologs are present in all land plants, including freezing-sensitive species, raising the question of SFR2 function beyond freezing tolerance. Similar to freezing, salt and drought can cause dehydration. Thus, it is hypothesized that in freezing-sensitive plants SFR2 may play roles in their resilience to salt or drought. To test this hypothesis, SlSFR2 RNAi lines were generated in the cold/freezing-sensitive species tomato (Solanum lycopersicum [M82 cv]). Hypersensitivity to salt and drought of SlSFR2-RNAi lines was observed. Higher tolerance of wild-type tomatoes was correlated with the production of trigalactosyldiacylglycerol, a product of SFR2 activity. Tomato SFR2 in vitro activity is Mg2+-dependent and its optimal pH is 7.5, similar to that of Arabidopsis SFR2, but the specific activity of tomato SFR2 in vitro is almost double that of Arabidopsis SFR2. When salt and drought stress were applied to Arabidopsis, no conditions could be identified at which SFR2 was induced prior to irreversibly impacting plant growth, suggesting that SFR2 protects Arabidopsis primarily against freezing. Discovery of tomato SFR2 function in drought and salt resilience provides further insights into general membrane lipid remodeling-based stress tolerance mechanisms and together with protection against freezing in freezing-resistant plants such as Arabidopsis, it adds lipid remodeling as a possible target for the engineering of abiotic stress-resilient crops. PMID:27600812

Arginine and proline applied as food additives stimulate high freeze tolerance in larvae of Drosophila melanogaster.

PubMed

Koštál, Vladimír; Korbelová, Jaroslava; Poupardin, Rodolphe; Moos, Martin; Šimek, Petr

2016-08-01

The fruit fly Drosophila melanogaster is an insect of tropical origin. Its larval stage is evolutionarily adapted for rapid growth and development under warm conditions and shows high sensitivity to cold. In this study, we further developed an optimal acclimation and freezing protocol that significantly improves larval freeze tolerance (an ability to survive at -5°C when most of the freezable fraction of water is converted to ice). Using the optimal protocol, freeze survival to adult stage increased from 0.7% to 12.6% in the larvae fed standard diet (agar, sugar, yeast, cornmeal). Next, we fed the larvae diets augmented with 31 different amino compounds, administered in different concentrations, and observed their effects on larval metabolomic composition, viability, rate of development and freeze tolerance. While some diet additives were toxic, others showed positive effects on freeze tolerance. Statistical correlation revealed tight association between high freeze tolerance and high levels of amino compounds involved in arginine and proline metabolism. Proline- and arginine-augmented diets showed the highest potential, improving freeze survival to 42.1% and 50.6%, respectively. Two plausible mechanisms by which high concentrations of proline and arginine might stimulate high freeze tolerance are discussed: (i) proline, probably in combination with trehalose, could reduce partial unfolding of proteins and prevent membrane fusions in the larvae exposed to thermal stress (prior to freezing) or during freeze dehydration; (ii) both arginine and proline are exceptional among amino compounds in their ability to form supramolecular aggregates which probably bind partially unfolded proteins and inhibit their aggregation under increasing freeze dehydration. © 2016. Published by The Company of Biologists Ltd.

Evidence of cyclical light/dark-regulated expression of freezing tolerance in young winter wheat plants.

PubMed

Skinner, Daniel Z; Bellinger, Brian; Hiscox, William; Helms, Gregory L

2018-01-01

The ability of winter wheat (Triticum aestivum L.) plants to develop freezing tolerance through cold acclimation is a complex rait that responds to many environmental cues including day length and temperature. A large part of the freezing tolerance is conditioned by the C-repeat binding factor (CBF) gene regulon. We investigated whether the level of freezing tolerance of 12 winter wheat lines varied throughout the day and night in plants grown under a constant low temperature and a 12-hour photoperiod. Freezing tolerance was significantly greater (P<0.0001) when exposure to subfreezing temperatures began at the midpoint of the light period, or the midpoint of the dark period, compared to the end of either period, with an average of 21.3% improvement in survival. Thus, freezing survival was related to the photoperiod, but cycled from low, to high, to low within each 12-hour light period and within each 12-hour dark period, indicating ultradian cyclic variation of freezing tolerance. Quantitative real-time PCR analysis of expression levels of CBF genes 14 and 15 indicated that expression of these two genes also varied cyclically, but essentially 180° out of phase with each other. Proton nuclear magnetic resonance analysis (1H-NMR) showed that the chemical composition of the wheat plants' cellular fluid varied diurnally, with consistent separation of the light and dark phases of growth. A compound identified as glutamine was consistently found in greater concentration in a strongly freezing-tolerant wheat line, compared to moderately and poorly freezing-tolerant lines. The glutamine also varied in ultradian fashion in the freezing-tolerant wheat line, consistent with the ultradian variation in freezing tolerance, but did not vary in the less-tolerant lines. These results suggest at least two distinct signaling pathways, one conditioning freezing tolerance in the light, and one conditioning freezing tolerance in the dark; both are at least partially under the control of the CBF regulon.

Comparison of Cryopreserved Human Sperm between Ultra Rapid Freezing and Slow Programmable Freezing: Effect on Motility, Morphology and DNA Integrity.

PubMed

Tongdee, Pattama; Sukprasert, Matchuporn; Satirapod, Chonticha; Wongkularb, Anna; Choktanasiri, Wicham

2015-05-01

Cryopreservation of sperm is common methods to preserve male fertility. Sperm freezing, suggest slow programmable freezing caused lower change of sperm morphology than sperm freezing in vapor of liquid nitrogen. Ultra rapid freezing is easy to be worked on, less time, low cost and does not need high experience. To compare the effect on sperm motility, morphology and DNA integrity of post-thawed sperm after ultra rapid freezing and slow programmable freezing methods. Experimental study at laboratory of infertility unit, Department of Obstetrics and Gynecology, Faculty of Medicine Ramathibodi Hospital. Thirty-seven semen samples with normal semen analysis according to World Health Organization (WHO) 1999 [normal sperm volume ( 2 ml) and normal sperm concentration (≥ 20 x10(6)/ml) and sperm motility (≥ 50%)]. Semen samples were washed. Then each semen sample was divided into six cryovials. Two cryovials, 0.5 ml each, were cryopreserved by slow programmable freezing. Four 0.25 ml containing cryovials, were cryopreserved by ultra rapidfreezing method. After cryopreservationfor 1 month, thawedprocess was carried out at room temperature. Main outcomes are sperm motility was determined by Computer-Assisted Semen Analysis (CASA), sperm morphology was determined by eosin-methylene blue staining and sperm DNA integrity was assessed by TUNEL assay. Sperm motility was reduced significantly by both methods, from 70.4 (9.0)% to 29.1 (12.3)% in slowprogrammable freezing and to 19.7 (9.8)% in ultra rapid freezing (p < 0.05). Sperm motility decreased significantly more by ultra rapid freezing (p < 0.001). The percentage of normal sperm morphology and DNA integrity were also reduced significantly by both methods. However, no significant difference between the two methods was found (p > 0.05). Cryopreservation of human sperm for 1 month significantly decreased sperm motility, morphology and DNA integrity in both methods. However sperm motility was decreased more by ultra rapid freezing.

A study of the impact of freezing on the lyophilization of a concentrated formulation with a high fill depth.

PubMed

Liu, Jinsong; Viverette, Todd; Virgin, Marlin; Anderson, Mitch; Paresh, Dalal

2005-01-01

The objective of this study was to evaluate the impact of freezing on the lyophilization of a concentrated formulation with a high fill depth. A model system consisting of a 15-mL fill of 15% (w/w) sulfobutylether 7-beta-cyclodextrin (SBECD) solution in a 30-mL vial was selected for this study. Various freezing methods including single-step freezing, two-step freezing with a super-cooling holding, annealing, vacuum-induced freezing, changing ice habit using tert-butyl-alcohol (TBA), ice nucleation with silver iodide (AgI), as well as combinations of some of the methods, were used in the lyophilization of this model system. This work demonstrated that the freezing process had a significant impact on primary drying rate and product quality of a concentrated formulation with a high fill depth. Annealing, vacuum-induced freezing, and addition of either TBA or an ice nucleating agent (AgI) to the formulation accelerated the subsequent ice sublimation process. Two-step freezing or addition of TBA improved the product quality by eliminating vertical heterogeneity within the cake. The combination of two-step freezing in conjunction with an annealing step was shown to be a method of choice for freezing in the lyophilization of a product with a high fill depth. In addition to being an effective method of freezing, it is most applicable for scaling up. An alternative approach is to add a certain amount of TBA to the formulation, if the TBA-formulation interaction or regulatory concerns can be demonstrated as not being an issue. An evaluation of vial size performed in this study showed that although utilizing large-diameter vials to reduce the fill depth can greatly shorten the cycle time of a single batch, it will substantially decrease the product throughput in a large-scale freeze-dryer.

A freeze-stable formulation for DTwP and DTaP vaccines.

PubMed

Xue, Honggang; Yang, Bangling; Kristensen, Debra D; Chen, Dexiang

2014-01-01

Inadvertent vaccine freezing often occurs in the cold chain and may cause damage to freeze‑sensitive vaccines. Liquid vaccines that contain aluminum salt adjuvants are particularly vulnerable. Polyol cryoprotective excipients have been shown to prevent freeze damage to hepatitis B vaccine. In this study, we examined the freeze-protective effect of propylene glycol on diphtheria-tetanus-pertussis-whole-cell (DTwP) and acellular (DTaP) vaccines. Pilot lots of DTwP and DTaP formulated with 7.5% propylene glycol underwent 3 freeze-thaw treatments. The addition of propylene glycol had no impact on pH, particle size distribution, or potency of the vaccines prior to freeze-thaw treatment; the only change noted was an increase in osmolality. The potencies and the physical properties of the vaccines containing cryoprotectant were maintained after freeze-thawing and for 3 months in accelerated stability studies. The results from this study indicate that formulating vaccines with propylene glycol can protect diphtheria-tetanus-pertussis vaccines against freeze damages.

Indirect contact freeze water desalination for an ice maker machine - CFD simulation

NASA Astrophysics Data System (ADS)

Jayakody, Harith; Al-Dadah, Raya; Mahmoud, Saad

2017-11-01

To offer for potable water shortages, sea water desalination is a potential solution for the global rising demand for fresh water. The latent heat of fusion is about one-seventh the latent heat of vaporisation, thus indicating the benefit of lower energy consumption for the freeze desalination process. Limited literature is reported on computational fluid dynamics (CFD) on freeze desalination. Therefore, analysing and investigating thermodynamic processes are easily conducted by the powerful tool of CFD. A single unit of ice formation in an ice maker machine was modelled using ANSYS Fluent software three-dimensionally. Energy, species transport and solidification/melting modules were used in building the CFD model. Parametric analysis was conducted using the established CFD model to predict the effects of freezing temperature and the geometry of the ice maker machine; on ice production and the freezing time. Lower freezing temperatures allowed more ice production and faster freezing. Increasing the diameter and the length of the freezing tube enabled more ice to be produced.

Freeze-drying of tert-butyl alcohol/water cosolvent systems: effects of formulation and process variables on residual solvents.

PubMed

Wittaya-Areekul, S; Nail, S L

1998-04-01

The objective of this study was to identify significant formulation and processing variables affecting levels of tert-butyl alcohol (TBA) and isopropyl alcohol (IPA) in freeze-dried solids prepared from TBA/water cosolvent systems. The variables examined were the physical state of the solute (crystalline vs amorphous), initial TBA concentration, freezing rate, cake thickness, and the temperature and duration of secondary drying. Sucrose and glycine were used as models for noncrystallizing and crystallizing solutes, respectively. The TBA concentration above which eutectic crystallization takes place was determined by differential scanning calorimetry. Model formulations were subjected to extremes of freezing rate by either dipping in liquid nitrogen or by slowly freezing on the shelf of a freeze-dryer. Dynamics of solvent loss during secondary drying was determined by withdrawing samples as a function of time at different shelf temperatures using a thief system. On the basis of these studies, the most important determinant of residual TBA level is the physical state of the solute. Freeze-dried glycine contained very low levels of residual TBA (0.01-0.03%) regardless of freezing rate or initial TBA concentration. For freeze-dried sucrose, residual TBA levels were approximately 2 orders of magnitude higher and were significantly affected by initial TBA concentration and freezing rate. For the sucrose/TBA/water system, relatively low residual TBA levels were obtained when the initial TBA level was above the threshold concentration for eutectic crystallization of TBA, whereas samples freeze-dried from solutions containing TBA concentrations below this threshold contained significantly higher levels of TBA. Residual IPA levels increased continuously with initial concentration of TBA in the sucrose/TBA/water system. Formulations of sucrose/TBA/water which were frozen rapidly contained residual TBA levels which were approximately twice those measured in the same formulation after slow freezing and drying under the same conditions. For the sucrose/TBA/water system, the temperature and time of secondary drying had only minimal influence on residual TBA in the freeze-dried solid. At low initial TBA concentrations (2%), residual TBA increases with increased cake thickness, perhaps because of the influence of depth of fill on effective freezing rate.

Characterization of freezing precipitation events through other meteorological variables and their recent changes over Northern Extratropics

NASA Astrophysics Data System (ADS)

Groisman, P. Y.; Yin, X.; Bulygina, O.

2017-12-01

Freezing precipitation events intertwine with agriculture, recreation, energy consumption, and seasonal transportation cycles of human activities. Using supplementary synoptic reports at 1,500 long-term stations of North America and Northern Eurasia, we created climatology of freezing precipitation near the surface and found significant changes (increases) in these occurrences in the past decade at high latitudes/elevations (Groisman et al. 2016; updated). Firstly, we document narrow boundaries of near surface temperature and humidity fields when freezing precipitation events occur; these are necessary but insufficient conditions of their occurrence. Secondly, using the upper air data at the sites collocated with in situ observations of freezing events, we quantify the typical pattern of lower troposphere temperature anomalies during freezing events: At the same locations and Julian days, the presence of freezing event at the surface is associated with significantly warmer temperatures in the lower troposphere; comparison of temperatures at nearest days before and after the freezing events with days during these events also shows statistically significant positive temperature anomalies in the lower troposphere to 500 hPa (on average, +3 to 4 °C) In the days with freezing events, vertical air temperature gradients between surface and 850 hPa become less than usual with frequent inversions, when the tropospheric air is warmer than at the surface. The above features of the lower tropospheric temperature, near-surface temperature and humidity represent a combination of weather conditions conducive for precipitation, if it happens, falling in the freezing rain form. The in situ reports of freezing events at synoptic stations allow us to estimate temporary and spatial distributions of such "special weather conditions". Thus, a posteriori high probability of freezing events under these weather conditions invokes similar probabilities of freezing rain over the ungauged terrain, where we do not have special synoptic reports but can reproduce these "special weather conditions" from less sophisticated observational networks and/or reanalyses. Reference: Groisman et al. 2016: Recent changes in the frequency of freezing precipitation in North America and Northern Eurasia. Environ Res Lett 11 045007.

Cryopreservation of Iberian pig spermatozoa. Comparison of different freezing extenders based on post-thaw sperm quality.

PubMed

De Mercado, Eduardo; Rodríguez, Ana; Gómez, Emilio; Sanz, Elena

2010-03-01

The aim of this study was to evaluate the cryoprotective effect of different freezing extenders against cryopreservation injuries on Iberian boar sperm. The sperm-rich fraction was collected and pooled from six sexually mature Iberian boars, and was frozen in different extenders containing glucose, lactose or fructose as sugar source and including Orvus ES Paste only in the freezing extender-2 (Glucose; Lactose and Fructose) or in both freezing extenders (Glucose2; Lactose2 and Fructose2). During the cryopreservation process, the supernatant was removed after the centrifugation step, then was extended with freezing extender-1 for the equilibration period and with freezing extender-2 immediately before freezing. Post-thaw sperm characteristics, such as plasma membrane integrity (SYBR-14/PI), mitochondrial function (Rhodamine 123) and acrosome integrity (NAR), were monitored. Overall sperm motility and the individual kinematic parameters of motile spermatozoa (assessed by the computer-aided sperm analysis system Sperm Class Analyzer [SCA]) were recorded in the different experimental treatments. Measurements were taken at 30 and 150 min post-thaw. The state of the acrosome after thawing did not show significant differences between the freezing extenders studied. Freezing-thawing caused a significant decrease (P<0.001) in plasma membrane integrity and in mitochondrial activity in the spermatozoa frozen with Orvus ES Paste in both freezing extenders. Furthermore, spermatozoa frozen with Orvus ES Paste in both freezing extenders exhibited lower (P<0.05) motility and kinematic parameters than those frozen in the absence of Orvus ES Paste in the first freezing extender. The spermatozoa frozen with the Lactose extender and with Orvus ES Paste only in the second freezing extender showed a better evolution of the motility and kinematic characteristics (P<0.05) over time. The deterioration in post-thaw sperm motility and kinematic parameters were concurrent with reduced sperm characteristics. It can be suggested that in the Iberian pig, the beneficial effects of Orvus ES Paste during the freezing process of spermatozoa is time dependent. The analysis of different sperm characteristics such as motility, plasma membrane integrity and mitochondrial function, determined that the extenders studied in the present experiment affected the quality of frozen-thawed semen in Iberian boar.

Care during freeze-drying of bovine pericardium tissue to be used as a biomaterial: a comparative study.

PubMed

Polak, Roberta; Pitombo, Ronaldo N M

2011-10-01

Bovine pericardium (BP) tissue is widely used in the manufacture of bioprosthetics. The effects of freeze-drying on the BP tissue have been studied by some researchers in order to decrease their cytotoxicity due to preservation in formaldehyde solution, and to increase the lifetime of the product in storage. This study was undertaken in order to study the effect of freeze-drying in the structure of BP. To perform this study BP samples were freeze-dried in two different types of freeze-dryers available in our laboratory: a laboratory freeze-dryer, in which it was not possible to control parameters and a pilot freeze-dryer, wherein all parameters during freezing and drying were controlled. After freeze-drying processes, samples were analyzed by SEM, Raman spectroscopy, tensile strength, water uptake tests and TEM. In summary, it has been demonstrated that damages occur in collagen fibers by the loss of bulk water of collagen structure implicating in a drastic decreasing of BP mechanical properties due to its structural alterations. Moreover, it was proven that the collagen fibrils suffered breakage at some points, which can be attributed to the uncontrolled parameters during drying. Copyright © 2011 Elsevier Inc. All rights reserved.

Perspective: Surface freezing in water: A nexus of experiments and simulations

NASA Astrophysics Data System (ADS)

Haji-Akbari, Amir; Debenedetti, Pablo G.

2017-08-01

Surface freezing is a phenomenon in which crystallization is enhanced at a vapor-liquid interface. In some systems, such as n-alkanes, this enhancement is dramatic and results in the formation of a crystalline layer at the free interface even at temperatures slightly above the equilibrium bulk freezing temperature. There are, however, systems in which the enhancement is purely kinetic and only involves faster nucleation at or near the interface. The first, thermodynamic, type of surface freezing is easier to confirm in experiments, requiring only the verification of the existence of crystalline order at the interface. The second, kinetic, type of surface freezing is far more difficult to prove experimentally. One material that is suspected of undergoing the second type of surface freezing is liquid water. Despite strong indications that the freezing of liquid water is kinetically enhanced at vapor-liquid interfaces, the findings are far from conclusive, and the topic remains controversial. In this perspective, we present a simple thermodynamic framework to understand conceptually and distinguish these two types of surface freezing. We then briefly survey fifteen years of experimental and computational work aimed at elucidating the surface freezing conundrum in water.

Three-Dimensional Microstructure of Biological Tissues during Freezing and Thawing

NASA Astrophysics Data System (ADS)

Ishiguro, Hiroshi; Horimizu, Takashi; Kataori, Akinobu; Kajigaya, Hiroshi

Three-dimensional behavior of ice crystals and cells during the freezing and thawing of biological tissues was investigated microscopically in real time by using a confocal laser scanning microscope(CLSM) and a fluorescent dye, acridine orange (AO). Fresh tender meat (2nd pectoral muscles) of chicken was stained with the AO in physiological saline to distinguish ice crystals and cells by their different colors, and then frozen and thawed under two different thermal protocols: a) slow-cooling and rapid-warming and b) rapid-cooling and rapid-warming. The CLSM noninvasively produced optical tomograms of the tissues to clarify the pattern of freezing, morphology of ice crystals in the tissues, and the interaction between ice crystals and cells. Also, the tissues were morphologically investigated by pathological means after the freezing and thawing. Typical freezing pattern during the slow-cooling was extracellular-freezing, and those during the rapid-cooling were extracellular-freezing and intracellular freezing with a lot of fine ice crystals in the cells. Cracks caused by the extracellular and intracellular ice crystals remained in the muscle tissues after the thawing. The results obtained by using the CLSM/dye method were consistent with pathologically morphological changes in the tissues through freezing and thawing.

Cold tolerance and freeze-induced glucose accumulation in three terrestrial slugs.

PubMed

Slotsbo, Stine; Hansen, Lars Monrad; Jordaens, Kurt; Backeljau, Thierry; Malmendal, Anders; Nielsen, Niels Chr; Holmstrup, Martin

2012-04-01

Cold tolerance and metabolic responses to freezing of three slug species common in Scandinavia (Arion ater, Arion rufus and Arion lusitanicus) are reported. Autumn collected slugs were cold acclimated in the laboratory and subjected to freezing conditions simulating likely winter temperatures in their habitat. Slugs spontaneously froze at about -4 °C when cooled under dry conditions, but freezing of body fluids was readily induced at -1 °C when in contact with external ice crystals. All three species survived freezing for 2 days at -1 °C, and some A. rufus and A. lusitanicus also survived freezing at -2 °C. (1)H NMR spectroscopy revealed that freezing of body fluids resulted in accumulation of lactate, succinate and glucose. Accumulation of lactate and succinate indicates that ATP production occurred via fermentative pathways, which is likely a result of oxygen depletion in frozen tissues. Glucose increased from about 6 to 22 μg/mg dry tissue upon freezing in A. rufus, but less so in A. ater and A. lusitanicus. Glucose may thus act as a cryoprotectant in these slugs, although the concentrations are not as high as reported for other freeze tolerant invertebrates. Copyright © 2012 Elsevier Inc. All rights reserved.

To freeze or not to freeze embryos: clarity, confusion and conflict.

PubMed

Goswami, Mohar; Murdoch, Alison P; Haimes, Erica

2015-06-01

Although embryo freezing is a routine clinical practice, there is little contemporary evidence on how couples make the decision to freeze their surplus embryos, or of their perceptions during that time. This article describes a qualitative study of 16 couples who have had in vitro fertilisation (IVF) treatment. The study question was 'What are the personal and social factors that patients consider when deciding whether to freeze embryos?' We show that while the desire for a baby is the dominant drive, couples' views revealed more nuanced and complex considerations in the decision-making process. It was clear that the desire to have a baby influenced couples' decision-making and that they saw freezing as 'part of the process'. However, there were confusions associated with the term 'freezing' related to concerns about the safety of the procedure. Despite being given written information, couples were confused about the practical aspects of embryo freezing, which suggests they were preoccupied with the immediate demands of IVF. Couples expressed ethical conflicts about freezing 'babies'. We hope the findings from this study will inform clinicians and assist them in providing support to couples confronted with this difficult decision-making.

Silicon-based Porous Ceramics via Freeze Casting of Preceramic Polymers

NASA Astrophysics Data System (ADS)

Naviroj, Maninpat

Freeze casting is a technique for processing porous materials that has drawn significant attention for its effectiveness in producing a variety of tailorable pore structures for ceramics, metals, and polymers. With freeze casting, pores are generated based on a solidification process where ice crystals act as a sacrificial template which can eventually be sublimated to create pores. While the majority of freeze-casting studies have been performed using conventional ceramic suspensions, this work explores an alternative processing route by freeze casting with preceramic polymer solutions. Significant differences exist between freeze casting of a particulate suspension and a polymeric solution. These changes affect the processing method, solidification behavior, and pore structure, thereby introducing new challenges and possibilities for the freeze-casting technique. The first part of this study explored the processing requirements involved with freeze casting of preceramic polymers, along with methods to control the resulting pore structure. Solvent choice, freezing front velocity, and polymer concentration were used as processing variables to manipulate the pore structures. A total of seven organic solvents were freeze cast with a polymethylsiloxane preceramic polymer to produce ceramics with isotropic, dendritic, prismatic, and lamellar pore morphologies. Changes in freezing front velocity and polymer concentration were shown to influence pore size, shape, and connectivity. Differences between suspension- and solution-based samples freeze cast under equivalent conditions were also investigated. Certain solidification microstructures were strongly affected by the presence of suspended particles, creating differences between pore structures generated from the same solvents. Additionally, processing of solution-based samples were found to be the more facile technique. Compressive strength and water permeability of dendritic and lamellar structures were analyzed to determine functional differences between the pore structures. Results show that dendritic structures were up to 30 times stronger, while lamellar structures provided higher permeability constants. A change in freezing front velocity was shown to significantly affect permeability but not compressive strength. Finally, improved pore alignment along the freezing direction was achieved by controlling the nucleation and growth of solvent crystals through the use of a grain-selection template. Dendritic samples freeze cast with a template showed substantial increase in pore alignment, as determined by image analysis and permeability tests, with the permeability constant increasing by up to 6-fold when compared to a control sample.

Modification of physical properties of freeze-dried rice

NASA Technical Reports Server (NTRS)

Huber, C. S.

1971-01-01

Freeze cycling process consists of alternately freezing and thawing precooked rice for two cycles, rice is then frozen and freeze-dehydrated in vacuum sufficient to remove water from rice by sublimitation. Process modifies rice grain structure and porosity, enabling complete rehydration in one minute in hot water.

9 CFR 590.534 - Freezing facilities.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Freezing facilities. 590.534 Section 590.534 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG..., and Facility Requirements § 590.534 Freezing facilities. (a) Freezing rooms, either on or off the...

9 CFR 590.536 - Freezing operations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Freezing operations. 590.536 Section 590.536 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG..., and Facility Requirements § 590.536 Freezing operations. (a) Freezing rooms shall be kept clean and...

The role of calcium and calmodulin in freezing-induced freezing resistance of Populus tomentosa cuttings.

PubMed

Lin, Shan-Zhi; Zhang, Zhi-Yi; Lin, Yuan-Zhen; Zhang, Qian; Guo, Huan

2004-02-01

To explore the role of calcium-calmodulin messenger system in the transduction of low temperature signal in woody plants, Populus tomentosa cuttings after being treated with CaCl(2) (10 mmol/L), Ca(2+) chelator EGTA (3 mmol/L), Ca(2+) channel inhibitor LaCl(3) (100 mmol/L) or CaM antagonist CPZ (50 mmol/L) were used for freezing acclimation at -3 degrees C. The changes in the calmodulin (CaM) and malonaldehyde (MDA) contents, the activities of superoxide dismutase (SOD), peroxidase (POD) and Ca(2+)-dependent adenosinetriphosphatase (Ca(2+)-ATPase) of mitochondrial membrane as well as freezing resistance (expressed as LT(50)) of cuttings were investigated to elucidate the physiological mechanisms by which trees adapt to freezing. The results showed that freezing acclimation increased the CaM content, the activities of SOD, POD and Ca(2+)-ATPase of mitochondrial membrane as well as freezing resistance of cuttings, and decreased the MDA content as compared with control cuttings. Treatment with CaCl(2) at the time of freezing acclimation enhanced the effect of freezing acclimation on the above-mentioned indexes, but this enhancement was abolished by Ca(2+)chelator EGTA, Ca(2+) channel inhibitor LaCl(3) or CaM antagonist CPZ, indicating that the calcium-calmodulin messenger system was involved in the course of freezing resistance development. The presence of CaCl(2) at the same time of freezing acclimation also reduced the degree of decline in CaM content, and in SOD, POD and Ca(2+)-ATPase activities caused by freezing stress at -14 degrees C, and enhanced the level of increase in CaM content, and in SOD, POD and Ca(2+)-ATPase activity in the recovery periods at 25 degrees C . The change in CaM content was found to be closely correlated to the levels of SOD, POD and Ca(2+)-ATPase, and to the degree of freezing resistance of cuttings during freezing acclimation either with or without CaCl(2) treatment. It was suggested that the increase of CaM content induced by CaCl(2) treatment promote the formation of Ca(2+)-CaM complexes, which effectively activates the activities of SOD, POD and mitochondrial Ca(2+)-ATPase and then further result in the adaptive changes associated with the development and enhancement of freezing resistance. Thus, It could be concluded that Ca(2+)-calmodulin may be involved in the regulation of the increase in SOD, POD and Ca(2+)-ATPase activities, and the induction of freezing resistance of cuttings.

Intact preservation of environmental samples by freezing under an alternating magnetic field.

PubMed

Morono, Yuki; Terada, Takeshi; Yamamoto, Yuhji; Xiao, Nan; Hirose, Takehiro; Sugeno, Masaya; Ohwada, Norio; Inagaki, Fumio

2015-04-01

The study of environmental samples requires a preservation system that stabilizes the sample structure, including cells and biomolecules. To address this fundamental issue, we tested the cell alive system (CAS)-freezing technique for subseafloor sediment core samples. In the CAS-freezing technique, an alternating magnetic field is applied during the freezing process to produce vibration of water molecules and achieve a stable, super-cooled liquid phase. Upon further cooling, the temperature decreases further, achieving a uniform freezing of sample with minimal ice crystal formation. In this study, samples were preserved using the CAS and conventional freezing techniques at 4, -20, -80 and -196 (liquid nitrogen) °C. After 6 months of storage, microbial cell counts by conventional freezing significantly decreased (down to 10.7% of initial), whereas that by CAS-freezing resulted in minimal. When Escherichia coli cells were tested under the same freezing conditions and storage for 2.5 months, CAS-frozen E. coli cells showed higher viability than the other conditions. In addition, an alternating magnetic field does not impact on the direction of remanent magnetization in sediment core samples, although slight partial demagnetization in intensity due to freezing was observed. Consequently, our data indicate that the CAS technique is highly useful for the preservation of environmental samples. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.

Freeze for action: neurobiological mechanisms in animal and human freezing

PubMed Central

2017-01-01

Upon increasing levels of threat, animals activate qualitatively different defensive modes, including freezing and active fight-or-flight reactions. Whereas freezing is a form of behavioural inhibition accompanied by parasympathetically dominated heart rate deceleration, fight-or-flight reactions are associated with sympathetically driven heart rate acceleration. Despite the potential relevance of freezing for human stress-coping, its phenomenology and neurobiological underpinnings remain largely unexplored in humans. Studies in rodents have shown that freezing depends on amygdala projections to the brainstem (periaqueductal grey). Recent neuroimaging studies in humans have indicated that similar brain regions may be involved in human freezing. In addition, flexibly shifting between freezing and active defensive modes is critical for adequate stress-coping and relies on fronto-amygdala connections. This review paper presents a model detailing these neural mechanisms involved in freezing and the shift to fight-or-flight action. Freezing is not a passive state but rather a parasympathetic brake on the motor system, relevant to perception and action preparation. Study of these defensive responses in humans may advance insights into human stress-related psychopathologies characterized by rigidity in behavioural stress reactions. The paper therefore concludes with a research agenda to stimulate translational animal–human research in this emerging field of human defensive stress responses. This article is part of the themed issue ‘Movement suppression: brain mechanisms for stopping and stillness’. PMID:28242739

Structurally Caused Freezing Point Depression of Biological Tissues

PubMed Central

Bloch, Rene; Walters, D. H.; Kuhn, Werner

1963-01-01

When investigating the freezing behaviour (by thermal analysis) of the glycerol-extracted adductor muscle of Mytilus edulis it was observed that the temperature of ice formation in the muscular tissue was up to 1.5°C lower than the freezing point of the embedding liquid, a 0.25 N KCl solution with pH = 4.9 with which the tissue had been equilibrated prior to the freezing experiment. A smaller freezing point depression was observed if the pH values of the embedding 0.25 N KCl solution were above or below pH = 4.9. Reasoning from results obtained previously in analogous experiments with artificial gels, the anomalous freezing depression is explained by the impossibility of growing at the normal freezing temperature regular macroscopic crystals inside the gel, due to the presence of the gel network. The freezing temperature is here determined by the size of the microprisms penetrating the meshes of the network at the lowered freezing temperature. This process leads finally to an ice block of more or less regular structure in which the filaments are embedded. Prerequisite for this hindrance of ideal ice growth is a sufficient tensile strength of the filamental network. The existence of structurally caused freezing point depression in biological tissue is likely to invalidate many conclusions reported in the literature, in which hypertonicity was deduced from cryoscopic data. PMID:13971682

Effects of buffer composition and processing conditions on aggregation of bovine IgG during freeze-drying.

PubMed

Sarciaux, J M; Mansour, S; Hageman, M J; Nail, S L

1999-12-01

The objective of this study was to identify critical formulation and processing variables affecting aggregation of bovine IgG during freeze-drying when no lyoprotective solute is used. Parameters examined were phosphate buffer concentration and counterion (Na versus K phosphate), added salts, cooling rate, IgG concentration, residual moisture level, and presence of a surfactant. No soluble aggregates were detected in any formulation after either freezing/thawing or freeze-drying. No insoluble aggregates were detected in any formulation after freezing, but insoluble aggregate levels were always detectable after freeze-drying. The data are consistent with a mechanism of aggregate formation involving denaturation of IgG at the ice/freeze-concentrate interface which is reversible upon freeze-thawing, but becomes irreversible after freeze-drying and reconstitution. Rapid cooling (by quenching in liquid nitrogen) results in more and larger aggregates than slow cooling on the shelf of the freeze-dryer. This observation is consistent with surface area measurements and environmental electron microscopic data showing a higher surface area of freeze-dried solids after fast cooling. Annealing of rapidly cooled solutions results in significantly less aggregation in reconstituted freeze-dried solids than in nonannealed controls, with a corresponding decrease in specific surface area of the freeze-dried, annealed system. Increasing the concentration of IgG significantly improves the stability of IgG against freeze-drying-induced aggregation, which may be explained by a smaller percentage of the protein residing at the ice/freeze-concentrate interface as IgG concentration is increased. A sodium phosphate buffer system consistently results in more turbid reconstituted solids than a potassium phosphate buffer system at the same concentration, but this effect is not attributable to a pH shift during freezing. Added salts such as NaCl or KCl contribute markedly to insoluble aggregate formation. Both sodium and potassium chloride contribute more to turbidity of the reconstituted solid than either sodium or potassium phosphate buffers at similar ionic strength, with sodium chloride resulting in a substantially higher level of aggregates than potassium chloride. At a given cooling rate, the specific surface area of dried solids is approximately a factor of 2 higher for the formulation containing sodium chloride than the formulation containing potassium chloride. Turbidity is also influenced by the extent of secondary drying, which underscores the importance of minimizing secondary drying of this system. Including a surfactant such as polysorbate 80, either in the formulation or in the water used for reconstitution, decreased, but did not eliminate, insoluble aggregates. There was no correlation between pharmaceutically acceptability of the freeze-dried cake and insoluble aggregate levels in the reconstituted product.

Wheeled and standard walkers in Parkinson's disease patients with gait freezing.

PubMed

Cubo, Esther; Moore, Charity G; Leurgans, Sue; Goetz, Christopher G

2003-10-01

Compare the efficacy of two walking assistance devices (wheeled walker and standard walker) to unassisted walking for patients with PD and gait freezing. Although numerous walking devices are used clinically, their relative effects on freezing and walking speed have never been systematically tested. Nineteen PD patients (14 non-demented) walked under three conditions in randomized order: unassisted walking, standard walker, and wheeled walker. Patients walked up to three times in each condition through a standard course that included rising from a chair, walking through a doorway, straightway walking, pivoting, and return. Total walking time, freezing time and number of freezes were compared for the three conditions using mixed models (walking time) and Friedman's test (freezing). The wheeled walker was further studied by comparing the effect of an attached laser that projected a bar of light on the floor as a visual walking cue. Use of either type of device significantly slowed walking compared to unassisted walking. Neither walker reduced any index of freezing, nor the laser attachment offered any advantage to the wheeled walker. The standard walker increased freezing, and the wheeled walker had no effect on freezing. Among the non-demented subjects (n=14), the same patterns occurred, although the walking speed was less impaired by the wheeled walker than the standard walker in this group. Though walkers may stabilize patients and increase confidence, PD patients walk more slowly when using them, without reducing freezing. Because the wheeled walker was intermediate for walking time and does not aggravate freezing, if walkers are used for these subjects, this type of walker should be favored.

Tracheid diameter is the key trait determining the extent of freezing-induced embolism in conifers.

PubMed

Pittermann, Jarmila; Sperry, John

2003-09-01

We tested the hypotheses that freezing-induced embolism is related to conduit diameter, and that conifers and angiosperms with conduits of equivalent diameter will exhibit similar losses of hydraulic conductivity in response to freezing. We surveyed the freeze-thaw response of conifers with a broad range of tracheid diameters by subjecting wood segments (root, stem and trunk wood) to a freeze-thaw cycle at -0.5 MPa in a centrifuge. Embolism increased as mean tracheid diameter exceeded 30 microm. Tracheids with a critical diameter greater than 43 microm were calculated to embolize in response to freezing and thawing at a xylem pressure of -0.5 MPa. To confirm that freezing-induced embolism is a function of conduit air content, we air-saturated stems of Abies lasiocarpa (Hook.) Nutt. (mean conduit diameter 13.7 +/- 0.7 microm) by pressurizing them 1 to 60 times above atmospheric pressure, prior to freezing and thawing. The air saturation method simulated the effect of increased tracheid size because the degree of super-saturation is proportional to a tracheid volume holding an equivalent amount of dissolved air at ambient pressure. Embolism increased when the dissolved air content was equivalent to a mean tracheid diameter of 30 microm at ambient air pressure. Our centrifuge and air-saturation data show that conifers are as vulnerable to freeze-thaw embolism as angiosperms with equal conduit diameter. We suggest that the hydraulic conductivity of conifer wood is maximized by increasing tracheid diameters in locations where freezing is rare. Conversely, the narrowing of tracheid diameters protects against freezing-induced embolism in cold climates.

Evaluation and Validation of the Messinger Freezing Fraction

NASA Technical Reports Server (NTRS)

Anderson, David N.; Tsao, Jen-Ching

2005-01-01

One of the most important non-dimensional parameters used in ice-accretion modeling and scaling studies is the freezing fraction defined by the heat-balance analysis of Messinger. For fifty years this parameter has been used to indicate how rapidly freezing takes place when super-cooled water strikes a solid body. The value ranges from 0 (no freezing) to 1 (water freezes immediately on impact), and the magnitude has been shown to play a major role in determining the physical appearance of the accreted ice. Because of its importance to ice shape, this parameter and the physics underlying the expressions used to calculate it have been questioned from time to time. Until now, there has been no strong evidence either validating or casting doubt on the current expressions. This paper presents experimental measurements of the leading-edge thickness of a number of ice shapes for a variety of test conditions with nominal freezing fractions from 0.3 to 1.0. From these thickness measurements, experimental freezing fractions were calculated and compared with values found from the Messinger analysis as applied by Ruff. Within the experimental uncertainty of measuring the leading-edge thickness, agreement of the experimental and analytical freezing fraction was very good. It is also shown that values of analytical freezing fraction were entirely consistent with observed ice shapes at and near rime conditions: At an analytical freezing fraction of unity, experimental ice shapes displayed the classic rime shape, while for conditions producing analytical freezing fractions slightly lower than unity, glaze features started to appear.

Multiple Glass Transitions and Freezing Events of Aqueous Citric Acid

PubMed Central

2014-01-01

Calorimetric and optical cryo-microscope measurements of 10–64 wt % citric acid (CA) solutions subjected to moderate (3 K/min) and slow (0.5 and 0.1 K/min) cooling/warming rates and also to quenching/moderate warming between 320 and 133 K are presented. Depending on solution concentration and cooling rate, the obtained thermograms show one freezing event and from one to three liquid–glass transitions upon cooling and from one to six liquid–glass and reverse glass–liquid transitions, one or two freezing events, and one melting event upon warming of frozen/glassy CA/H2O. The multiple freezing events and glass transitions pertain to the mother CA/H2O solution itself and two freeze-concentrated solution regions, FCS1 and FCS2, of different concentrations. The FCS1 and FCS2 (or FCS22) are formed during the freezing of CA/H2O upon cooling and/or during the freezing upon warming of partly glassy or entirely glassy mother CA/H2O. The formation of two FCS1 and FCS22 regions during the freezing upon warming to our best knowledge has never been reported before. Using an optical cryo-microscope, we are able to observe the formation of a continuous ice framework (IF) and its morphology and reciprocal distribution of IF/(FCS1 + FCS2). Our results provide a new look at the freezing and glass transition behavior of aqueous solutions and can be used for the optimization of lyophilization and freezing of foods and biopharmaceutical formulations, among many other applications where freezing plays a crucial role. PMID:25482069

Freeze Tolerance of Nine Zoysiagrass Cultivars Using Natural Cold Acclimation and Freeze Chambers

USDA-ARS?s Scientific Manuscript database

Winter hardiness of zoysiagrass (Zoysia spp.) cultivars is an important attribute throughout the biogeographical transition zone, thus the inability to withstand freezing temperatures may limit the use of these cultivars. The objective of this research was to determine the freeze tolerance (LT50) of...

Surface freezing of water.

PubMed

Pérez-Díaz, J L; Álvarez-Valenzuela, M A; Rodríguez-Celis, F

2016-01-01

Freezing, melting, evaporation and condensation of water are essential ingredients for climate and eventually life on Earth. In the present work, we show how surface freezing of supercooled water in an open container is conditioned and triggered-exclusively-by humidity in air. Additionally, a change of phase is demonstrated to be triggered on the water surface forming surface ice crystals prior to freezing of bulk. The symmetry of the surface crystal, as well as the freezing point, depend on humidity, presenting at least three different types of surface crystals. Humidity triggers surface freezing as soon as it overpasses a defined value for a given temperature, generating a plurality of nucleation nodes. An evidence of simultaneous nucleation of surface ice crystals is also provided.

An Investigation of the Cryogenic Freezing of Water in Non-Metallic Pipelines

NASA Astrophysics Data System (ADS)

Martin, C. I.; Richardson, R. N.; Bowen, R. J.

2004-06-01

Pipe freezing is increasingly used in a range of industries to solve otherwise intractable pipe line maintenance and servicing problems. This paper presents the interim results from an experimental study on deliberate freezing of polymeric pipelines. Previous and contemporary works are reviewed. The object of the current research is to confirm the feasibility of ice plug formation within a polymeric pipe as a method of isolation. Tests have been conducted on a range of polymeric pipes of various sizes. The results reported here all relate to freezing of horizontal pipelines. In each case the process of plug formation was photographed, the frozen plug pressure tested and the pipe inspected for signs of damage resulting from the freeze procedure. The time to freeze was recorded and various temperatures logged. These tests have demonstrated that despite the poor thermal and mechanical properties of the polymers, freezing offers a viable alternative method of isolation in polymeric pipelines.

Frost heave susceptibility of saturated soil under constant rate of freezing

NASA Astrophysics Data System (ADS)

Ryokai, K.; Iguro, M.; Yoneyama, K.

Introduced are the results of experiments carried out to quantitatively obtain the frost heave pressure and displacement of soil subjected to artificial freezing or freezing around in-ground liquefied natural gas storage tanks. This experiment is conducted to evaluate the frost heave susceptibility of saturated soil under overconsolidation. In other words, this experiment was carried out to obtain the relation of the over-burden pressure and freezing rate to the frost heave ratio by observing the frost heave displacement and freezing time of specimens by freezing the specimens at a constant freezing rate under a constant overburden pressure, while letting water freely flow in and out of the system. Introduced are the procedures for frost heave test required to quantitatively obtain the frost heave displacement and pressure of soil. Furthermore, the relation between the frost heave susceptibility and physical properties of soil obtained by this test is reported.

Contact Freezing of Water by Salts.

PubMed

Niehaus, Joseph; Cantrell, Will

2015-09-03

Water is unlikely to crystallize homogeneously at temperatures greater than -34 °C. Freezing at higher temperatures is heterogeneous-catalyzed by the presence of a second substance. If that substance is at an air-water interface, then the mode is called contact freezing, and it typically will trigger nucleation at a higher temperature than if the substance were wholly immersed within the liquid. We find that the impact of salt particles initiates freezing in experiments using water droplets at supercoolings of 9 to 16 °C. These results show that contact freezing nuclei need not be effective as immersion mode nuclei. We discuss our results in the context of proposed mechanisms of contact freezing. Finally, we use the time scales for diffusion of heat and of ions and the propagation of a sound wave through the droplet to estimate that contact freezing occurs within 10 ns of impact.

Cold hardiness in molluscs

NASA Astrophysics Data System (ADS)

Ansart, Armelle; Vernon, Philippe

2003-05-01

Molluscs inhabit all types of environments: seawater, intertidal zone, freshwater and land, and of course may have to deal with subzero temperatures. Ectotherm animals survive cold conditions by avoiding it by extensive supercooling (freezing avoidant species) or by bearing the freezing of their extracellular body fluids (freezing tolerant species). Although some studies on cold hardiness are available for intertidal molluscs, they are scarce for freshwater and terrestrial ones. Molluscs often exhibit intermediary levels of cold hardiness, with a moderate or low ability to supercool and a limited survival to the freezing of their tissues. Several factors could be involved: their dependence on water, their ability to enter dormancy, the probability of inoculative freezing in their environment, etc. Size is an important parameter in the development of cold hardiness abilities: it influences supercooling ability in land snails, which are rather freezing avoidant and survival to ice formation in intertidal organisms, which generally tolerate freezing.

Effects of industrial pre-freezing processing and freezing handling on glucosinolates and antioxidant attributes in broccoli florets.

PubMed

Cai, Congxi; Miao, Huiying; Qian, Hongmei; Yao, Leishuan; Wang, Bingliang; Wang, Qiaomei

2016-11-01

The effects of industrial pre-freezing processing and freezing handling on the contents of glucosinolates and antioxidants (vitamin C, polyphenols, carotenoid and chlorophyll), as well as the antioxidant capacity in broccoli (Brassica oleracea L. var. italica) florets were investigated in the present study. Our results showed that the glucosinolate accumulations were significantly decreased after pre-freezing processing, whereas elevated levels of phenols, carotenoids, chlorophyll, and also antioxidant capacity were observed in frozen broccoli florets. The contents of vitamin C remained constant during above mentioned processing. In conclusion, the current industrial freezing processing method is a good practice for the preservation of main antioxidant nutrients in broccoli florets, although some improvements in pre-freezing processing, such as steam blanching and ice-water cooling, are needed to attenuate the decrease in glucosinolate content. Copyright © 2016 Elsevier Ltd. All rights reserved.

Two-dimensional freezing criteria for crystallizing colloidal monolayers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang Ziren; Han Yilong; Alsayed, Ahmed M.

Video microscopy was employed to explore crystallization of colloidal monolayers composed of diameter-tunable microgel spheres. Two-dimensional (2D) colloidal liquids were frozen homogenously into polycrystalline solids, and four 2D criteria for freezing were experimentally tested in thermal systems for the first time: the Hansen-Verlet freezing rule, the Loewen-Palberg-Simon dynamical freezing criterion, and two other rules based, respectively, on the split shoulder of the radial distribution function and on the distribution of the shape factor of Voronoi polygons. Importantly, these freezing criteria, usually applied in the context of single crystals, were demonstrated to apply to the formation of polycrystalline solids. At themore » freezing point, we also observed a peak in the fluctuations of the orientational order parameter and a percolation transition associated with caged particles. Speculation about these percolated clusters of caged particles casts light on solidification mechanisms and dynamic heterogeneity in freezing.« less

Effect of pasteurisation and freezing method on bioactive compounds and antioxidant activity of strawberry pulp.

PubMed

Gonçalves, Gilma Auxiliadora Santos; Resende, Nathane Silva; Carvalho, Elisângela Elena Nunes; Resende, Jaime Vilela de; Vilas Boas, Eduardo Valério de Barros

2017-09-01

This study evaluated the stability of strawberry pulp subjected to three factors, pasteurisation (pasteurised and unpasteurised), freezing method (static air and forced air) and storage time (0, 2, 4 and 6 months). Pasteurisation favoured vitamin C retention during storage but enhanced the total loss of phenolics without affecting anthocyanin levels. Freezing by forced air was more effective in retaining phenolics during the first 4 months of storage, although the freezing method did not affect the anthocyanin levels. Processing and storage reduced the levels of individual phenolics. Freezing by forced air was more effective than static air in retaining antioxidant activity of the pulp. Polyphenol oxidase and peroxidase enzyme levels were relatively stable and independent of pasteurisation, freezing and storage time. Even after 6 months of frozen storage, strawberry pulp is a significant source of nutrients and bioactive compounds and retains high antioxidant capacity independent of pasteurisation and freezing method.

Freezing and thawing or freezing, thawing, and aging effects on beef tenderness

USDA-ARS?s Scientific Manuscript database

The objective of this study was to determine the effect of freezing and thawing or freezing and thawing with an additional aging period after frozen storage on the tenderness of longissimus lumborum (LL) and semitendinosus (ST) steaks relative to aged, fresh steaks. Left-side LL and ST (n=35 each) ...

7 CFR 929.11 - To can, freeze, or dehydrate.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 8 2010-01-01 2010-01-01 false To can, freeze, or dehydrate. 929.11 Section 929.11... LONG ISLAND IN THE STATE OF NEW YORK Order Regulating Handling Definitions § 929.11 To can, freeze, or dehydrate. To can, freeze, or dehydrate means to convert cranberries into canned, frozen, or dehydrated...

Applications of Simulator Freeze to Carrier Glideslope Tracking Instruction.

DTIC Science & Technology

1982-07-01

Showing Datum Bars and Meatball . .. .. .. ... .. ... .... 19 4 Freezes Per Trial Averaged Across Freeze Conditions and Across 4-Trial Blocks of Training...algorithm linearly increased the criterion in meatball units from 1.0 at 6000 feet from the ramp to 1.5 at the ramp. "Freezes" did not occur beyond 6000

7 CFR 305.18 - Quick freeze treatment schedule.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 5 2010-01-01 2010-01-01 false Quick freeze treatment schedule. 305.18 Section 305.18... SERVICE, DEPARTMENT OF AGRICULTURE PHYTOSANITARY TREATMENTS Quick Freeze Treatments § 305.18 Quick freeze treatment schedule. (a) T110. (1) Initially, lower the commodity's temperature to 0 °F or below. (2) Hold...

The equilibrated state of freezing as a basis for distinguishing lethal stresses of freezing in plants

USDA-ARS?s Scientific Manuscript database

A model for coordination of stresses that limit winterhardiness in plants based on the thermodynamic equilibrated state of freezing and melting provides a rational basis for distinction of freeze-induced energies which can stress and injure living organisms in various ways. The departure from equili...

7 CFR 58.620 - Freezing and packaging rooms.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 3 2013-01-01 2013-01-01 false Freezing and packaging rooms. 58.620 Section 58.620... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Rooms and Compartments § 58.620 Freezing and packaging rooms. The rooms used for freezing and packaging frozen desserts shall be...

7 CFR 58.620 - Freezing and packaging rooms.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 3 2014-01-01 2014-01-01 false Freezing and packaging rooms. 58.620 Section 58.620... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Rooms and Compartments § 58.620 Freezing and packaging rooms. The rooms used for freezing and packaging frozen desserts shall be...

7 CFR 58.620 - Freezing and packaging rooms.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 3 2012-01-01 2012-01-01 false Freezing and packaging rooms. 58.620 Section 58.620... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Rooms and Compartments § 58.620 Freezing and packaging rooms. The rooms used for freezing and packaging frozen desserts shall be...

Freezing temperature of finger skin.

PubMed

Wilson, O; Goldman, R F; Molnar, G W

1976-10-01

In 45 subjects, 154 frostnips of the finger were induced by cooling in air at -15 degrees C with various wind speeds. The mean supercooled skin temperature at which frostnip appeared was -9.4 degrees C. The mean skin temperature rise due to heat of fusion at ice crystallization was 5.3 degrees C. The skin temperature rose to what was termed the apparent freezing point. The relation of this point to the supercooled skin temperature was analyzed for the three wind speeds used. An apparent freezing point for a condition of no supercooling was calculated, estimating the highest temperature at which skin freezes at a given wind speed. The validity of the obtained differences in apparent freezing point was tested by an analysis of covariance. Although not statistically significant, the data suggest that the apparent freezing point with no supercooling decreases with increasing wind velocity. The highest calculated apparent freezing point at -15 degrees C and 6.8 m/s was 1.2 degrees C lower than the true freezing point for skin previously determined in brine, which is a statistically significant difference.

A Vivens Ex Vivo Study on the Synergistic Effect of Electrolysis and Freezing on the Cell Nucleus.

PubMed

Lugnani, Franco; Zanconati, Fabrizio; Marcuzzo, Thomas; Bottin, Cristina; Mikus, Paul; Guenther, Enric; Klein, Nina; Rubinsky, Liel; Stehling, Michael K; Rubinsky, Boris

2015-01-01

Freezing-cryosurgery, and electrolysis-electrochemical therapy (EChT), are two important minimally invasive surgery tissue ablation technologies. Despite major advantages they also have some disadvantages. Cryosurgery cannot induce cell death at high subzero freezing temperatures and requires multiple freeze thaw cycles, while EChT requires high concentrations of electrolytic products-which makes it a lengthy procedure. Based on the observation that freezing increases the concentration of solutes (including products of electrolysis) in the frozen region and permeabilizes the cell membrane to these products, this study examines the hypothesis that there could be a synergistic effect between freezing and electrolysis in their use together for tissue ablation. Using an animal model we refer to as vivens ex vivo, which may be of value in reducing the use of animals for experiments, combined with a Hematoxylin stain of the nucleus, we show that there are clinically relevant protocols in which the cell nucleus appears intact when electrolysis and freezing are used separately but is affected by certain combinations of electrolysis and freezing.

Inner ear tissue preservation by rapid freezing: Improving fixation by high-pressure freezing and hybrid methods

PubMed Central

Bullen, A.; Taylor, R.R.; Kachar, B.; Moores, C.; Fleck, R.A.; Forge, A.

2014-01-01

In the preservation of tissues in as ‘close to life’ state as possible, rapid freeze fixation has many benefits over conventional chemical fixation. One technique by which rapid freeze-fixation can be achieved, high pressure freezing (HPF), has been shown to enable ice crystal artefact-free freezing and tissue preservation to greater depths (more than 40 μm) than other quick-freezing methods. Despite increasingly becoming routine in electron microscopy, the use of HPF for the fixation of inner ear tissue has been limited. Assessment of the quality of preservation showed routine HPF techniques were suitable for preparation of inner ear tissues in a variety of species. Good preservation throughout the depth of sensory epithelia was achievable. Comparison to chemically fixed tissue indicated that fresh frozen preparations exhibited overall superior structural preservation of cells. However, HPF fixation caused characteristic artefacts in stereocilia that suggested poor quality freezing of the actin bundles. The hybrid technique of pre-fixation and high pressure freezing was shown to produce cellular preservation throughout the tissue, similar to that seen in HPF alone. Pre-fixation HPF produced consistent high quality preservation of stereociliary actin bundles. Optimising the preparation of samples with minimal artefact formation allows analysis of the links between ultrastructure and function in inner ear tissues. PMID:25016142

Identification and classification of genes required for tolerance to freeze-thaw stress revealed by genome-wide screening of Saccharomyces cerevisiae deletion strains.

PubMed

Ando, Akira; Nakamura, Toshihide; Murata, Yoshinori; Takagi, Hiroshi; Shima, Jun

2007-03-01

Yeasts used in bread making are exposed to freeze-thaw stress during frozen-dough baking. To clarify the genes required for freeze-thaw tolerance, genome-wide screening was performed using the complete deletion strain collection of diploid Saccharomyces cerevisiae. The screening identified 58 gene deletions that conferred freeze-thaw sensitivity. These genes were then classified based on their cellular function and on the localization of their products. The results showed that the genes required for freeze-thaw tolerance were frequently involved in vacuole functions and cell wall biogenesis. The highest numbers of gene products were components of vacuolar H(+)-ATPase. Next, the cross-sensitivity of the freeze-thaw-sensitive mutants to oxidative stress and to cell wall stress was studied; both of these are environmental stresses closely related to freeze-thaw stress. The results showed that defects in the functions of vacuolar H(+)-ATPase conferred sensitivity to oxidative stress and to cell wall stress. In contrast, defects in gene products involved in cell wall assembly conferred sensitivity to cell wall stress but not to oxidative stress. Our results suggest the presence of at least two different mechanisms of freeze-thaw injury: oxidative stress generated during the freeze-thaw process, and defects in cell wall assembly.

Application of freeze-drying technology in manufacturing orally disintegrating films.

PubMed

Liew, Kai Bin; Odeniyi, Michael Ayodele; Peh, Kok-Khiang

2016-01-01

Freeze drying technology has not been maximized and reported in manufacturing orally disintegrating films. The aim of this study was to explore the freeze drying technology in the formulation of sildenafil orally disintegrating films and compare the physical properties with heat-dried orally disintegrating film. Central composite design was used to investigate the effects of three factors, namely concentration of carbopol, wheat starch and polyethylene glycol 400 on the tensile strength and disintegration time of the film. Heat-dried films had higher tensile strength than films prepared using freeze-dried method. For folding endurance, freeze-dried films showed improved endurance than heat-dried films. Moreover, films prepared using freeze-dried methods were thicker and had faster disintegration time. Formulations with higher amount of carbopol and starch showed higher tensile strength and thickness whereas formulations with higher PEG 400 content showed better flexibility. Scanning electron microscopy showed that the freeze-dried films had more porous structure compared to the heat-dried film as a result of the release of water molecule from the frozen structure when it was subjected to freeze drying process. The sildenafil film was palatable. The dissolution profiles of freeze-dried and heat-dried films were similar to Viagra® with f2 of 51.04 and 65.98, respectively.

Freezing: an underutilized food safety technology?

PubMed

Archer, Douglas L

2004-01-15

Freezing is an ancient technology for preserving foods. Freezing halts the activities of spoilage microorganisms in and on foods and can preserve some microorganisms for long periods of time. Frozen foods have an excellent overall safety record. The few outbreaks of food-borne illness associated with frozen foods indicate that some, but not all human pathogens are killed by commercial freezing processes. Freezing kills microorganisms by physical and chemical effects and possibly through induced genetic changes. Research is needed to better understand the physical and chemical interactions of various food matrices with the microbial cell during freezing and holding at frozen temperatures. The literature suggests that many pathogenic microorganisms may be sublethally injured by freezing, so research should be done to determine how to prevent injured cells from resuscitating and becoming infectious. Studies on the genetics of microbial stress suggest that the induction of resistance to specific stresses may be counteracted by, for example, simple chemicals. Research is needed to better understand how resistance to the lethal effect of freezing is induced in human pathogens and means by which it can be counteracted in specific foods. Through research, it seems possible that freezing may in the future be used to reliably reduce populations of food-borne pathogens as well as to preserve foods.

The Use of High-resolution Infrared Thermography (HRIT) for the Study of Ice Nucleation and Ice Propagation in Plants

PubMed Central

Wisniewski, Michael; Neuner, Gilbert; Gusta, Lawrence V.

2015-01-01

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events often have devastating effects on agricultural production and can also play an important role in shaping community structure in natural populations of plants, especially in alpine, sub-arctic, and arctic ecosystems. Therefore, a better understanding of the freezing process in plants can play an important role in the development of methods of frost protection and understanding mechanisms of freeze avoidance. Here, we describe a protocol to visualize the freezing process in plants using high-resolution infrared thermography (HRIT). The use of this technology allows one to determine the primary sites of ice formation in plants, how ice propagates, and the presence of ice barriers. Furthermore, it allows one to examine the role of extrinsic and intrinsic nucleators in determining the temperature at which plants freeze and evaluate the ability of various compounds to either affect the freezing process or increase freezing tolerance. The use of HRIT allows one to visualize the many adaptations that have evolved in plants, which directly or indirectly impact the freezing process and ultimately enables plants to survive frost events. PMID:25992743

The use of high-resolution infrared thermography (HRIT) for the study of ice nucleation and ice propagation in plants.

PubMed

Wisniewski, Michael; Neuner, Gilbert; Gusta, Lawrence V

2015-05-08

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events often have devastating effects on agricultural production and can also play an important role in shaping community structure in natural populations of plants, especially in alpine, sub-arctic, and arctic ecosystems. Therefore, a better understanding of the freezing process in plants can play an important role in the development of methods of frost protection and understanding mechanisms of freeze avoidance. Here, we describe a protocol to visualize the freezing process in plants using high-resolution infrared thermography (HRIT). The use of this technology allows one to determine the primary sites of ice formation in plants, how ice propagates, and the presence of ice barriers. Furthermore, it allows one to examine the role of extrinsic and intrinsic nucleators in determining the temperature at which plants freeze and evaluate the ability of various compounds to either affect the freezing process or increase freezing tolerance. The use of HRIT allows one to visualize the many adaptations that have evolved in plants, which directly or indirectly impact the freezing process and ultimately enables plants to survive frost events.

Water-cooled probe technique for the study of freeze lining formation

NASA Astrophysics Data System (ADS)

Verscheure, Karel; Campforts, Mieke; Verhaeghe, Frederik; Boydens, Eddy; Blanpain, Bart; Wollants, Patrick; van Camp, Maurits

2006-12-01

Furnace protection by water-cooled freeze linings becomes increasingly important as the metal producing industry attempts to achieve higher process intensities. Systematic investigations of the growth and the resulting microstructure and compositional profile of freeze linings are necessary to understand the behavior of freeze linings, their relation with the industrial process, and their interaction with the wall cooling system. We have developed a technique based on the submergence of a water-cooled probe into a liquid slag bath. Freeze linings of two industrial nonferrous slags have been produced using this technique and their growth, microstructural, and compositional profiles as a function of submergence time were determined. Thermodynamic equilibrium for the investigated slag systems was calculated and compared with the observed microstructures. The freeze linings form in approximately 15 minutes. Close to the water cooling, the freeze linings are predominantly amorphous in structure. With increasing distance from the water cooling, the proportion of crystalline phases increases and bath material is entrapped in the microstructure. Cellular crystals are observed close to the bath. The freeze linings exhibit an approximate homogeneous composition. The results demonstrate that the technique is a successful tool in obtaining information on the growth, microstructure, and composition of freeze linings in industrial water-cooled furnaces.

Freeze-dried spermatozoa: A future tool?

PubMed

Olaciregui, M; Gil, L

2017-04-01

Cryopreservation has been routinely used to preserve sperm of human and different animal species. However, frozen sperm storage for a long time brings many inconveniences because of liquid nitrogen. Many attempts have been made to overcome the disadvantages of the current cryopreservation method. Freeze-drying has been proposed as alternative method for sperm preservation to achieve the ability to store sperm doses indefinitely at ambient temperature or in ordinary refrigerators. At present, it has been reported successfully sperm freeze-drying on many animal species including canine and feline. It is well known that during freeze-drying process, sperm DNA could be damaged, but if suitable protection is provided, the sperm nucleus could preserve the ability to activate the oocyte and embryos could be generated by intracytoplasmic sperm injection (ICSI). Many factors influence the freeze-drying efficacy, so current researches have been conducted to find strategies to control these factors to maintain the sperm DNA integrity. This review describes the latest method of sperm freeze-drying for practical application in preserving and transporting genetic resources. In addition, the approaches to improve the efficiency of the technique were studied. We demonstrated that the DNA integrity of freeze-dried dog sperm is affected by the composition of the freeze-drying solution as well as the temperature and period of storage. Further studies are necessary to refine freeze-drying protocol in order to protect the DNA and maintain the sperm functionality and obtain offspring from freeze-dried sperm. © 2016 Blackwell Verlag GmbH.

Role of cells in freezing-induced cell-fluid-matrix interactions within engineered tissues.

PubMed

Seawright, Angela; Ozcelikkale, Altug; Dutton, Craig; Han, Bumsoo

2013-09-01

During cryopreservation, ice forms in the extracellular space resulting in freezing-induced deformation of the tissue, which can be detrimental to the extracellular matrix (ECM) microstructure. Meanwhile, cells dehydrate through an osmotically driven process as the intracellular water is transported to the extracellular space, increasing the volume of fluid for freezing. Therefore, this study examines the effects of cellular presence on tissue deformation and investigates the significance of intracellular water transport and cell-ECM interactions in freezing-induced cell-fluid-matrix interactions. Freezing-induced deformation characteristics were examined through cell image deformetry (CID) measurements of collagenous engineered tissues embedded with different concentrations of MCF7 breast cancer cells versus microspheres as their osmotically inactive counterparts. Additionally, the development of a biophysical model relates the freezing-induced expansion of the tissue due to the cellular water transport and the extracellular freezing thermodynamics for further verification. The magnitude of the freezing-induced dilatation was found to be not affected by the cellular water transport for the cell concentrations considered; however, the deformation patterns for different cell concentrations were different suggesting that cell-matrix interactions may have an effect. It was, therefore, determined that intracellular water transport during freezing was insignificant at the current experimental cell concentrations; however, it may be significant at concentrations similar to native tissue. Finally, the cell-matrix interactions provided mechanical support on the ECM to minimize the expansion regions in the tissues during freezing.

Alcohol Brine Freezing of Japanese Horse Mackerel (Trachurus japonicus) for Raw Consumption

NASA Astrophysics Data System (ADS)

Maeda, Toshimichi; Yuki, Atsuhiko; Sakurai, Hiroshi; Watanabe, Koichiro; Itoh, Nobuo; Inui, Etsuro; Seike, Kazunori; Mizukami, Yoichi; Fukuda, Yutaka; Harada, Kazuki

In order to test the possible application of alcohol brine freezing to Japanese horse mackerel (Trachurus japonicus) for raw consumption, the quality and taste of fish frozen by direct immersion in 60% ethanol brine at -20, -25 and -30°C was compared with those by air freezing and fresh fish without freezing. Cracks were not found during the freezing. Smell of ethanol did not remain. K value, an indicator of freshness, of fish frozen in alcohol brine was less than 8.3%, which was at the same level as those by air freezing and fresh fish. Oxidation of lipid was at the same level as air freezing does, and lower than that of fresh fish. The pH of fish frozen in alcohol brine at -25 and -30°C was 6.5 and 6.6, respectively, which were higher than that by air freezing and that of fresh fish. Fish frozen in alcohol brine was better than that by air and at the same level as fresh fish in total evaluation of sensory tests. These results show that the alcohol brine freezing is superior to air freezing, and fish frozen in alcohol brine can be a material for raw consumption. The methods of thawing in tap water, cold water, refrigerator, and at room temperature were compared. Thawing in tap water is considered to be convenient due to the short thaw time and the quality of thawed fish that was best among the methods.

Contribution of extracellular ice formation and the solution effects to the freezing injury of PC-3 cells suspended in NaCl solutions.

PubMed

Takamatsu, Hiroshi; Zawlodzka, Sylwia

2006-08-01

The mechanism of cell injury during slow freezing was examined using PC-3 human prostate adenocarcinoma cells suspended in NaCl solutions. The objective was to evaluate contribution of extracellular ice and the 'solution effects' to freezing injury separately. The solution effects that designate the influence of elevated concentration were evaluated from a pseudo-freezing experiment, where cells were subjected to the milieu that simulated a freeze-thaw process by changing the NaCl concentration and the temperature at the same time. The effect of extracellular ice formation on cell injury was then estimated from the difference in cell survival between the pseudo-freezing experiment and a corresponding freezing experiment. When cells were frozen to a relatively higher freezing temperature at -10 degrees C, about 30% of cells were damaged mostly due to extracellular ice formation, because the concentration increase without ice formation to 2.5-M NaCl, i.e., the equilibrium concentration at -10 degrees C, had no effect on cell survival. In contrast, in the case of the lower freezing temperature at -20 degrees C, about 90% of cells were injured by both effects, particularly 60-80% by the solution effects among them. The present results suggested that the solution effects become more crucial to cell damage during slow freezing at lower temperatures, while the effect of ice is limited to some extent.

Effects of freezing conditions on quality changes in blueberries.

PubMed

Cao, Xuehui; Zhang, Fangfang; Zhao, Dongyu; Zhu, Danshi; Li, Jianrong

2018-03-12

Freezing preservation is one of the most effective methods used to maintain the flavour and nutritional value of fruit. This research studied the effects of different freezing conditions, -20 °C, -40 °C, -80 °C, and immersion in liquid nitrogen, on quality changes of freeze-thawed blueberries. The water distribution estimates of blueberries were measured based on low-field nuclear magnetic resonance (LF-NMR) analysis. The pectin content, drip loss, and fruit texture were also detected to evaluate quality changes in samples. The freezing curves of blueberry showed super-cooling points at -20 °C and - 40 °C, whereas super-cooling points were not observed at -80 °C or in liquid nitrogen. After freeze-thaw treatment, the relaxation time of the cell wall water (T 21 ), cytoplasm water and extracellular space (T 22 ), and vacuole water (T 23 ) were significantly shortened compared to fresh samples, which suggested a lower liquidity. Although the freezing speed for samples immersed in liquid nitrogen was faster than other treatments, samples treated at -80 °C showed better quality regarding vacuole water holding, drip loss, and original pectin content retention. This study contributed to understanding how freezing temperature affects the qualities of blueberries. The super-fast freezing rate might injure fruit, and an appropriate freezing rate could better preserve blueberries. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

Involvement of Multiple Types of Dehydrins in the Freezing Response in Loquat (Eriobotrya japonica)

PubMed Central

Xu, Hongxia; Yang, Yong; Xie, Li; Li, Xiaoying; Feng, Chao; Chen, Junwei; Xu, Changjie

2014-01-01

Dehydrins (DHNs) are a family of plant proteins typically induced in response to stress conditions that cause cellular dehydration, such as low temperatures, high salinity, and drought. Loquat (Eriobotrya japonica) is a perennial fruit crop that blossoms during winter. Loquat fruitlets are frequently injured by freezing. To evaluate the role of the EjDHNs in freezing resistance in loquat fruitlets, two cultivars of loquat, the freezing-sensitive ‘Ninghaibai’ (FS-NHB) and the freezing-tolerant ‘Jiajiao’ (FT-JJ), were analyzed under induced freezing stress. Freezing stress led to obvious accumulation of reactive oxygen species and considerable lipid peroxidation in membranes during the treatment period. Both these phenomena were more pronounced in ‘FS-NHB’ than in ‘FS-JJ.’ Immunogold labeling of dehydrin protein was performed. DHN proteins were found to be concentrated mainly in the vicinity of the plasma membrane, and the density of the immunogold labeling was significantly higher after freezing treatment, especially in the more freezing-tolerant cultivar ‘FT-JJ.’ Seven DHNs, showing four different structure types, were obtained from loquat fruitlets and used to study the characteristics of different EjDHN proteins. These DHN proteins are all highly hydrophilic, but they differ significantly in size, ranging from 188 to 475 amino acids, and in biochemical properties, such as theoretical pI, aliphatic index, and instability index. Freezing treatment resulted in up-regulation of the expression levels of all seven EjDHNs, regardless of structure type. The accumulation of the transcripts of these EjDHN genes was much more pronounced in ‘FT-JJ’ than in ‘FS-NHB.’ Altogether, this study provides evidence that EjDHNs are involved in the cryoprotection of the plasma membrane during freeze-induced dehydration in loquat fruitlets. PMID:24498141

Osmotically Induced Reversible Transitions in Lipid-DNA Mesophases

PubMed Central

Danino, Dganit; Kesselman, Ellina; Saper, Gadiel; Petrache, Horia I.; Harries, Daniel

2009-01-01

We follow the effect of osmotic pressure on isoelectric complexes that self-assemble from mixtures of DNA and mixed neutral and cationic lipids. Using small angle x-ray diffraction and freeze-fracture cryo-electron microscopy, we find that lamellar complexes known to form in aqueous solutions can reversibly transition to hexagonal mesophases under high enough osmotic stress exerted by adding a neutral polymer. Using molecular spacings derived from x-ray diffraction, we estimate the reversible osmotic pressure-volume (Π-V) work needed to induce this transition. We find that the transition free energy is comparable to the work required to elastically bend lipid layers around DNA. Consistent with this, the required work is significantly lowered by an addition of hexanol, which is known to soften lipid bilayers. Our findings not only help to resolve the free-energy contributions associated with lipid-DNA complex formation, but they also demonstrate the importance that osmotic stress can have to the macromolecular phase geometry in realistic biological environments. PMID:19348739

Nuclear Involvement in the Appearance of a Chloroplast-Encoded 32,000 Dalton Thylakoid Membrane Polypeptide Integral to the Photosystem II Complex 1

PubMed Central

Leto, Kenneth J.; Keresztes, Aron; Arntzen, Charles J.

1982-01-01

The genetic locus for the high chlorophyll fluorescent photosystem II-deficient maize mutant hcf*-3 has been definitively located to the nuclear genome. Fluorography of lamellar polypeptides labeled with [35S]methionine in vivo revealed the specific loss of a heavily labeled 32,000 dalton thylakoid membrane polypeptide as well as its chloroplast encoded precursor species at 34,000 daltons. Examination of freeze-fractured mesophyll and bundle sheath thylakoids from hcf*-3 revealed that both plastid types lacked the large EFs particles believed to consist of the photosystem II reaction center-core complex and associated light harvesting chlorophyll-proteins. The present evidence suggests that the synthesis or turnover/integration of the chloroplast-encoded 34,000 to 32,000 dalton polypeptide is under nuclear control, and that these polyipeptides are integral components of photosystem II which may be required for the assembly or structural stabilization of newly formed photosystem II reaction centers in both mesophyll and bundle sheath chloroplasts. Images PMID:16662421

Analysis of Workplace Accidents in Automotive Repair Workshops in Spain.

PubMed

López-Arquillos, Antonio; Rubio-Romero, Juan Carlos

2016-09-01

To analyze the effects of the factors associated with different types of injury (superficial wounds, dislocations and sprains, bone fractures, concussion and internal injuries, burns scalding and freezing) caused by occupational accidents in automotive repair workshops. Study of a sample consisting of 89,954 industry accidents reported from 2003 to 2008. Odds ratios were calculated with a 95% confidence interval. Belonging to a small company is a risk factor for suffering three of the five types of injury studied. Women are less likely to suffer burns and superficial wounds, and more likely to suffer dislocations or sprains. Foreign workers are more likely to suffer concussion and internal injuries. Health and safety strategies and accident prevention measures should be individualized and adapted to the type of worker most likely to be injured in each type of accident. Occupational health and safety training courses designed according to worker profile, and improving the participation of the workers in small firms creating regional or roving safety representatives would improve working conditions.

Single-Pixel Optical Fluctuation Analysis of Calcium Channel Function in Active Zones of Motor Nerve Terminals

PubMed Central

Luo, Fujun; Dittrich, Markus; Stiles, Joel R.; Meriney, Stephen D.

2011-01-01

We used high-resolution fluorescence imaging and single-pixel optical fluctuation analysis to estimate the opening probability of individual voltage-gated calcium (Ca2+) channels during an action potential and the number of such Ca2+ channels within active zones of frog neuromuscular junctions. Analysis revealed ~36 Ca2+ channels within each active zone, similar to the number of docked synaptic vesicles but far less than the total number of transmembrane particles reported based on freeze-fracture analysis (~200–250). The probability that each channel opened during an action potential was only ~0.2. These results suggest why each active zone averages only one quantal release event during every other action potential, despite a substantial number of docked vesicles. With sparse Ca2+ channels and low opening probability, triggering of fusion for each vesicle is primarily controlled by Ca2+ influx through individual Ca2+ channels. In contrast, the entire synapse is highly reliable because it contains hundreds of active zones. PMID:21813687

Environmental Scanning Electron Microscope Imaging of Vesicle Systems.

PubMed

Perrie, Yvonne; Ali, Habib; Kirby, Daniel J; Mohammed, Afzal U R; McNeil, Sarah E; Vangala, Anil

2017-01-01

The structural characteristics of liposomes have been widely investigated and there is certainly a strong understanding of their morphological characteristics. Imaging of these systems, using techniques such as freeze-fracturing methods, transmission electron microscopy, and cryo-electron imaging, has allowed us to appreciate their bilayer structures and factors which can influence this. However, there are few methods which all us to study these systems in their natural hydrated state; commonly the liposomes are visualized after drying, staining, and/or fixation of the vesicles. Environmental Scanning Electron Microscopy (ESEM) offers the ability to image a liposome in its hydrated state without the need for prior sample preparation. Within our studies we were the first to use ESEM to study liposomes and niosomes and we have been able to dynamically follow the hydration of lipid films and changes in liposome suspensions as water condenses on to, or evaporates from, the sample in real time. This provides insight into the resistance of liposomes to coalescence during dehydration, thereby providing an alternative assay of liposome formulation and stability.

Functional analysis of tight junction organization.

PubMed

DiBona, D R

1985-01-01

The functional basis of tight junction design has been examined from the point of view that this rate-limiting barrier to paracellular transport is a multicompartment system. Review of the osmotic sensitivity of these structures points to the need for this sort of analysis for meaningful correlation of structure and function under a range of conditions. A similar conclusion is drawn with respect to results from voltage-clamping protocols where reversal of spontaneous transmural potential difference elicits parallel changes in both structure and function in much the same way as does reversal of naturally occurring osmotic gradients. In each case, it becomes necessary to regard the junction as a functionally polarized structure to account for observations of its rectifying properties. Lastly, the details of experimentally-induced junction deformation are examined in light of current theories of its organization; arguments are presented in favor of the view that the primary components of intramembranous organization (as viewed with freeze-fracture techniques) are lipidic rather than proteinaceous.

Antimicrobial efficiency of essential oil and freeze-thaw treatments against Escherichia coli O157:H7 and Salmonella enterica Ser. Enteritidis in strawberry juice.

PubMed

Duan, J; Zhao, Y

2009-04-01

This study investigated the antimicrobial efficiency of 3 essential oils (EOs), lemongrass, cinnamon leaf, and basil, and freeze-thaw treatment, alone or in combination, against Escherichia coli O157:H7 and Salmonella enterica Ser. Enteritidis inoculated in strawberry juice stored at 7 degrees C. EO of lemongrass or cinnamon leaf at 0.1 to 2 microL/mL and freezing at -23 degrees C for 24 or 48 h followed by thawing at 7 degrees C for 4 h all showed significant antimicrobial activities (P < 0.05) against E. coli O157:H7 and S. Enteritidis in strawberry juice. The antimicrobial activity increased with increasing EO concentration and storage time, but extending freezing time from 24 to 48 h did not enhance the antimicrobial activity of freeze-thaw treatment (P > 0.05). EO of lemongrass or cinnamon leaf at 0.1 microL/mL and freeze-thaw treatment alone obtained a 5 log(10) reduction in the population of S. Enteritidis, while EOs at 0.1 to 0.3 microL/mL or freeze-thaw alone could not achieve a satisfactory protection against E. coli O157:H7 in strawberry juice. Combined EO and freeze-thaw treatment enhanced the overall antimicrobial effect against E. coli O157:H7, with adding EO before the freeze-thaw treatment showed a faster decontamination rate than when added EO after the freeze-thaw. EOs of lemongrass and cinnamon leaf at 0.1 or 0.3 microL/mL followed by the freeze-thawing resulted in a 5 log(10) reduction in E. coli O157:H7 on the 5th and 2nd day of storage, respectively. This study suggested that combined EO and freeze-thaw treatment may be a suitable and inexpensive method to eliminate microorganisms that can be a hazard for the consumers of unpasteurized berry juices.

"On" freezing in Parkinson's disease: resistance to visual cue walking devices.

PubMed

Kompoliti, K; Goetz, C G; Leurgans, S; Morrissey, M; Siegel, I M

2000-03-01

To measure "on" freezing during unassisted walking (UW) and test if two devices, a modified inverted stick (MIS) and a visual laser beam stick (LBS) improved walking speed and number of "on" freezing episodes in patients with Parkinson's disease (PD). Multiple visual cues can overcome "off' freezing episodes and can be useful in improving gait function in parkinsonian patients. These devices have not been specifically tested in "on" freezing, which is unresponsive to pharmacologic manipulations. Patients with PD, motor fluctuations and freezing while "on," attempted walking on a 60-ft track with each of three walking conditions in a randomized order: UW, MIS, and LBS. Total time to complete a trial, number of freezes, and the ratio of walking time to the number of freezes were compared using Friedman's test. Twenty-eight patients with PD, mean age 67.81 years (standard deviation [SD] 7.54), mean disease duration 13.04 years (SD 7.49), and mean motor Unified Parkinson's Disease Rating Scale score "on" 32.59 (SD 10.93), participated in the study. There was a statistically significant correlation of time needed to complete a trial and number of freezes for all three conditions (Spearman correlations: UW 0.973, LBS 0.0.930, and MIS 0.842). The median number of freezes, median time to walk in each condition, and median walking time per freeze were not significantly different in pairwise comparisons of the three conditions (Friedman's test). Of the 28 subjects, six showed improvement with the MIS and six with the LBS in at least one outcome measure. Assisting devices, specifically based on visual cues, are not consistently beneficial in overcoming "on" freezing in most patients with PD. Because this is an otherwise untreatable clinical problem and because occasional subjects do respond, cautious trials of such devices under the supervision of a health professional should be conducted to identify those patients who might benefit from their long-term use.

Freezing of gait in PD: prospective assessment in the DATATOP cohort.

PubMed

Giladi, N; McDermott, M P; Fahn, S; Przedborski, S; Jankovic, J; Stern, M; Tanner, C

2001-06-26

To study the development of freezing of gait in PD. Freezing of gait is a common, disabling, and poorly understood symptom in PD. The authors analyzed data from 800 patients with early PD from the Deprenyl and Tocopherol Antioxidative Therapy of Parkinsonism (DATATOP) clinical trial who were assigned either placebo, deprenyl, tocopherol, or the combination of deprenyl and tocopherol. The primary outcome measure was the time from randomization until the freezing of gait score on the Unified Parkinson's Disease Rating Scale (UPDRS) became positive. Fifty-seven patients (7.1%) had freezing of gait at study entry and 193 (26%) of the remaining patients experienced the symptom by the end of the follow-up period. Those with freezing of gait at baseline had significantly more advanced disease than those without the symptom, as measured by total UPDRS and Hoehn and Yahr stage. High baseline risk factors for developing freezing of gait during the follow-up period were the onset of PD with a gait disorder; higher scores of rigidity, postural instability, bradykinesia and speech; and longer disease duration. In contrast, tremor was strongly associated with a decreased risk for freezing of gait. At the end of follow-up, the signs most strongly associated with the freezing phenomenon were gait, balance, and speech disorders, not rigidity or bradykinesia. Deprenyl treatment was strongly associated with a decreased risk for developing freezing of gait; tocopherol had no effect. Freezing of gait is directly related to duration of PD. Risk factors at onset of disease are the absence of tremor and PD beginning as a gait disorder. The development of freezing of gait in the course of the illness is strongly associated with the development of balance and speech problems, less so with the worsening of bradykinesia, and is not associated with the progression of rigidity. These results support the concept that the freezing phenomenon is distinct from bradykinesia. Deprenyl, in the absence of L-dopa, was found to be an effective prophylactic treatment and should be considered for patients with PD who have an onset of gait difficulty.

Gelatin-Filtered Consomme: A Practical Demonstration of the Freezing and Thawing Processes

ERIC Educational Resources Information Center

Lahne, Jacob B.; Schmidt, Shelly J.

2010-01-01

Freezing is a key food processing and preservation technique widely used in the food industry. Application of best freezing and storage practices extends the shelf-life of foods for several months, while retaining much of the original quality of the fresh food. During freezing, as well as its counterpart process, thawing, a number of critical…

The 2007 California Citrus Freeze: Vulnerability, Poverty, and Unemployment Issues of Farmworkers

ERIC Educational Resources Information Center

Orozco, Graciela Leon

2010-01-01

In January of 2007, freezing temperatures destroyed citrus and vegetable crops in California. By May 2007, more than 9,000 freeze-related unemployment applications had been fled. Through face-to-face interviews, this study documents the experience of 63 farmworkers to find out how they survived the freeze and accessed services. Findings revealed…

Method of freezing living cells and tissues with improved subsequent survival

DOEpatents

Senkan, Selim M.; Hirsch, Gerald P.

1980-01-01

This invention relates to an improved method for freezing red blood cells, ther living cells, or tissues with improved subsequent survival, wherein constant-volume freezing is utilized that results in significantly improved survival compared with constant-pressure freezing; optimization is attainable through the use of different vessel geometries, cooling baths and warming baths, and sample concentrations.

21 CFR 864.9225 - Cell-freezing apparatus and reagents for in vitro diagnostic use.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Cell-freezing apparatus and reagents for in vitro... Establishments That Manufacture Blood and Blood Products § 864.9225 Cell-freezing apparatus and reagents for in vitro diagnostic use. (a) Identification. Cell-freezing apparatus and reagents for in vitro diagnostic...

21 CFR 864.9225 - Cell-freezing apparatus and reagents for in vitro diagnostic use.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Cell-freezing apparatus and reagents for in vitro... Establishments That Manufacture Blood and Blood Products § 864.9225 Cell-freezing apparatus and reagents for in vitro diagnostic use. (a) Identification. Cell-freezing apparatus and reagents for in vitro diagnostic...

Plant moisture stress: a portable freezing-point meter compared with the psychrometer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cary, J.W.; Fisher, H.D.

A small portable instrument for measuring the freezing-point depression of plant tissue has been developed for field use. The instrument is easy to operate and can be constructed from materials costing less than $100. Moisture stress measurements made with the freezing-point meter on a variety of plants were compared with vapor pressure psychrometer measurements. Variation between duplicates in the freezing point averaged 1.2 bars, but differences between stress measurements made with the psychrometer and freezing-point instrument averaged 2.6 bars. 11 references, 5 figures, 2 tables.

Freeze drying apparatus

DOEpatents

Coppa, Nicholas V.; Stewart, Paul; Renzi, Ernesto

2001-01-01

The present invention provides methods and apparatus for freeze drying in which a solution, which can be a radioactive salt dissolved within an acid, is frozen into a solid on vertical plates provided within a freeze drying chamber. The solid is sublimated into vapor and condensed in a cold condenser positioned above the freeze drying chamber and connected thereto by a conduit. The vertical positioning of the cold condenser relative to the freeze dryer helps to help prevent substances such as radioactive materials separated from the solution from contaminating the cold condenser. Additionally, the system can be charged with an inert gas to produce a down rush of gas into the freeze drying chamber to also help prevent such substances from contaminating the cold condenser.

Freeze drying method

DOEpatents

Coppa, Nicholas V.; Stewart, Paul; Renzi, Ernesto

1999-01-01

The present invention provides methods and apparatus for freeze drying in which a solution, which can be a radioactive salt dissolved within an acid, is frozen into a solid on vertical plates provided within a freeze drying chamber. The solid is sublimated into vapor and condensed in a cold condenser positioned above the freeze drying chamber and connected thereto by a conduit. The vertical positioning of the cold condenser relative to the freeze dryer helps to help prevent substances such as radioactive materials separated from the solution from contaminating the cold condenser. Additionally, the system can be charged with an inert gas to produce a down rush of gas into the freeze drying chamber to also help prevent such substances from contaminating the cold condenser.

Selectivity in biomineralization of barium and strontium.

PubMed

Krejci, Minna R; Wasserman, Brian; Finney, Lydia; McNulty, Ian; Legnini, Daniel; Vogt, Stefan; Joester, Derk

2011-11-01

The desmid green alga Closterium moniliferum belongs to a small number of organisms that form barite (BaSO(4)) or celestite (SrSO(4)) biominerals. The ability to sequester Sr in the presence of an excess of Ca is of considerable interest for the remediation of (90)Sr from the environment and nuclear waste. While most cells dynamically regulate the concentration of the second messenger Ca(2+) in the cytosol and various organelles, transport proteins rarely discriminate strongly between Ca, Sr, and Ba. Herein, we investigate how these ions are trafficked in C. moniliferum and how precipitation of (Ba,Sr)SO(4) crystals occurs in the terminal vacuoles. Towards this goal, we simultaneously visualize intracellular dynamics of multiple elements using X-ray fluorescence microscopy (XFM) of cryo-fixed/freeze-dried samples. We correlate the resulting elemental maps with ultrastructural information gleaned from freeze-fracture cryo-SEM of frozen-hydrated cells and use micro X-ray absorption near edge structure (micro-XANES) to determine sulfur speciation. We find that the kinetics of Sr uptake and efflux depend on external Ca concentrations, and Sr, Ba, and Ca show similar intracellular localization. A highly ion-selective cross-membrane transport step is not evident. Based on elevated levels of sulfate detected in the terminal vacuoles, we propose a "sulfate trap" model, where the presence of dissolved barium leads to preferential precipitation of (Ba,Sr)SO(4) due to its low solubility relative to SrSO(4) and CaSO(4). Engineering the sulfate concentration in the vacuole may thus be the most direct way to increase the Sr sequestered per cell, an important consideration in using desmids for phytoremediation of (90)Sr. Copyright © 2011 Elsevier Inc. All rights reserved.

[Preparation of freeze-dried powder of recombinate hirudin-2 nanoparticle for nasal delivery and permeability through nasal membrane in vitro].

PubMed

Chen, Ming-Xia; Zhang, Jian-Bao; Yu, Ji-Ping; Ye, Jing; Wei, Bao-Hong; Zhang, Yu-Jie

2013-06-01

To optimize the freeze-dried powder preparation technology of recombinate hirudin-2 (rHV2) nanoparticle which has bio-adhesive characteristic for nasal delivery, also to investigate its stability and permeability through nasal membrane in vitro. Taking the appearance, rediffusion of nanoparticle and rHV2 encapsulation efficiency as the evaluation indexes. Cryoprotector, the preparative technique and the effect of illumination and high temperature factors on its stability for rHV2 freeze-dried powder were investigated. Using Fraze diffusion cell technique, the permeability of rHV2 across rabbit nasal mucous membrane in chitosan solution, chitosan nanoparticle, and nanoparticle frozen-dried powder were compared with that in normal saline solution. The optimized preparation of rHV2 nanoparticle freeze-dried powder was as follows: 5% trehalose and glucose (1:1) was used as cryoprotector, nanoparticle solution was freezed for 24 h in vacuum frozen-dryer after being pre-freezed for 24 h. The content of rHV2 in the freeze-dried powder was 1.1 ug/mg. Illumination had little effect on the appearance, rediffusion and encapsulation efficiency of the rHV2 freeze-dried powder. High temperature could obviously influence the appearance of nanoparticle freeze-dried powder. The permeability coefficient (P) of nanoparticle was 5 times more than that in chictonson solution. It was indicated that chitosan nanoparticle has effect on increasing the permeability of rHV2. The freeze-dried powder of chitosan nanoparticle can be a good nasal preparation of rHV2.

Effect of freeze-drying on viability and in vitro probiotic properties of a mixture of lactic acid bacteria and yeasts isolated from kefir.

PubMed

Bolla, Patricia A; Serradell, María de los Angeles; de Urraza, Patricio J; De Antoni, Graciela L

2011-02-01

The effect of freeze-drying on viability and probiotic properties of a microbial mixture containing selected bacterial and yeast strains isolated from kefir grains (Lactobacillus kefir, Lactobacillus plantarum, Lactococcus lactis, Saccharomyces cerevisiae and Kluyveromyces marxianus) was studied. The microorganisms were selected according to their potentially probiotic properties in vitro already reported. Two types of formulations were performed, a microbial mixture (MM) suspended in milk and a milk product fermented with MM (FMM). To test the effect of storage on viability of microorganisms, MM and FMM were freeze-dried and maintained at 4°C for six months. After 180 days of storage at 4°C, freeze-dried MM showed better survival rates for each strain than freeze-dried FMM. The addition of sugars (trehalose or sucrose) did not improve the survival rates of any of the microorganisms after freeze-drying. Freeze-drying did not affect the capacity of MM to inhibit growth of Shigella sonnei in vitro, since the co-incubation of this pathogen with freeze-dried MM produced a decrease of 2 log in Shigella viability. The safety of freeze-dried MM was tested in mice and non-translocation of microorganisms to liver or spleen was observed in BALB/c mice feed ad libitum during 7 or 20 days. To our knowledge, this is the first report about the effect of freeze-drying on viability, in vitro probiotic properties and microbial translocation of a mixture containing different strains of both bacteria and yeasts isolated from kefir.

Additional freeze hardiness in wheat acquired by exposure to -3 degreesC is associated with extensive physiological, morphological, and molecular changes.

PubMed

Herman, Eliot M; Rotter, Kelsi; Premakumar, Ramaswamy; Elwinger, G; Bae, Hanhong; Bae, Rino; Ehler-King, Linda; Chen, Sixue; Livingston, David P

2006-01-01

Cold-acclimated plants acquire an additional 3-5 degrees C increase in freezing tolerance when exposed to -3 degrees C for 12-18 h before a freezing test (LT50) is applied. The -3 degrees C treatment replicates soil freezing that can occur in the days or weeks leading to overwintering by freezing-tolerant plants. This additional freezing tolerance is called subzero acclimation (SZA) to differentiate it from cold acclimation (CA) that is acquired at above-freezing temperatures. Using wheat as a model, results have been obtained indicating that SZA is accompanied by changes in physiology, cellular structure, the transcriptome, and the proteome. Using a variety of assays, including DNA arrays, reverse transcription-polymerase chain reaction (RT-PCR), 2D gels with mass spectroscopic identification of proteins, and electron microscopy, changes were observed to occur as a consequence of SZA and the acquisition of added freezing tolerance. In contrast to CA, SZA induced the movement of intracellular water to the extracellular space. Many unknown and stress-related genes were upregulated by SZA including some with obvious roles in SZA. Many genes related to photosynthesis and plastids were downregulated. Changes resulting from SZA often appeared to be a loss of rather than an appearance of new proteins. From a cytological perspective, SZA resulted in alterations of organelle structure including the Golgi. The results indicate that the enhanced freezing tolerance of SZA is correlated with a wide diversity of changes, indicating that the additional freezing tolerance is the result of complex biological processes.

Soil Moisture Controls the Thermal Habitat of Active Layer Soils in the McMurdo Dry Valleys, Antarctica

NASA Astrophysics Data System (ADS)

Wlostowski, A. N.; Gooseff, M. N.; Adams, B. J.

2018-01-01

Antarctic soil ecosystems are strongly controlled by abiotic habitat variables. Regional climate change in the McMurdo Dry Valleys is expected to cause warming over the next century, leading to an increase in frequency of freeze-thaw cycling in the soil habitat. Previous studies show that physiological stress associated with freeze-thaw cycling adversely affects invertebrate populations by decreasing abundance and positively selecting for larger body sizes. However, it remains unclear whether or not climate warming will indeed enhance the frequency of annual freeze-thaw cycling and associated physiological stresses. This research quantifies the frequency, rate, and spatial heterogeneity of active layer freezing to better understand how regional climate change may affect active layer soil thermodynamics, and, in turn, affect soil macroinvertebrate communities. Shallow active layer temperature, specific conductance, and soil moisture were observed along natural wetness gradients. Field observations show that the frequency and rate of freeze events are nonlinearly related to freezable soil moisture (θf). Over a 2 year period, soils at θf < 0.080 m3/m3 experienced between 15 and 35 freeze events and froze rapidly compared to soils with θf > 0.080 m3/m3, which experienced between 2 and 6 freeze events and froze more gradually. A numerical soil thermodynamic model is able to simulate observed freezing rates across a range of θf, reinforcing a well-known causal relationship between soil moisture and active layer freezing dynamics. Findings show that slight increases in soil moisture can potentially offset the effect of climate warming on exacerbating soil freeze-thaw cycling.

Detailed spectral profile analysis of electrocorticograms during freezing against penicillin-induced epileptiform discharges in the anesthetized rat.

PubMed

Tokiwa, Tatsuji; Zimin, Lev; Inoue, Takao; Nomura, Sadahiro; Suzuki, Michiyasu; Yamakawa, Takeshi

2018-07-01

Cryosurgery is an alternative technique for minimally invasive treatment of lesions. We have recently examined cryosurgery for epilepsy in animal models, and found that penicillin G (PG)-induced epileptiform discharges (EDs) mostly vanished after freezing. However, EDs were provoked again after insufficient freezing. Inadequate freezing is not visually detectable during and just after freezing and is not predictable beforehand. To manage this problem, we examined whether intraoperative monitoring of electrocorticograms (ECoGs) can predict recurrence of EDs after cryosurgery. A palm-sized cryoprobe system was applied to focal seizures in a Wistar rat model in which EDs were induced in advance by intracerebral injection of PG. During stable induction of EDs, the cryoprobe was carefully inserted into the epileptic region and this region was immediately frozen. After the series of prefreezing, freezing, and postfreezing, rats in which PG-induced EDs relapsed within 3 h were defined as the ED-relapsed (EDR) group, and other rats were defined as the ED-vanished (EDV) group. Time-frequency analysis was conducted on the ECoGs in each group through each freezing series. Relapse of PG-induced EDs on ECoG after the freezing series was associated with the remaining power of the delta band in the freezing period more strongly in the EDR group than in the EDV group. Success or failure of the freezing procedure can be predicted by the specificity of the delta band of the ECoG obtained intraoperatively. Copyright © 2018 Elsevier B.V. All rights reserved.

Thermomechanical analysis of freezing-induced cell-fluid-matrix interactions in engineered tissues

PubMed Central

Han, Bumsoo; Teo, Ka Yaw; Ghosh, Soham; Dutton, J. Craig; Grinnell, Frederick

2012-01-01

Successful cryopreservation of functional engineered tissues (ETs) is significant to tissue engineering and regenerative medicine, but it is extremely challenging to develop a successful protocol because the effects of cryopreservation parameters on the post-thaw functionality of ETs are not well understood. Particularly, the effects on the microstructure of their extracellular matrix (ECM) have not been well studied, which determines many functional properties of the ETs. In this study, we investigated the effects of two key cryopreservation parameters – i) freezing temperature and corresponding cooling rate; and ii) the concentration of cryoprotective agent (CPA) on the ECM microstructure as well as the cellular viability. Using dermal equivalent as a model ET and DMSO as a model CPA, freezing-induced spatiotemporal deformation and post-thaw ECM microstructure of ETs was characterized while varying the freezing temperature and DMSO concentrations. The spatial distribution of cellular viability and the cellular actin cytoskeleton was also examined. The results showed that the tissue dilatation increased significantly with reduced freezing temperature (i.e., rapid freezing). A maximum limit of tissue deformation was observed for preservation of ECM microstructure, cell viability and cell-matrix adhesion. The dilatation decreased with the use of DMSO, and a freezing temperature dependent threshold concentration of DMSO was observed. The threshold DMSO concentration increased with lowering freezing temperature. In addition, an analysis was performed to delineate thermodynamic and mechanical components of freezing-induced tissue deformation. The results are discussed to establish a mechanistic understanding of freezing-induced cell-fluid-matrix interaction and phase change behavior within ETs in order to improve cryopreservation of ETs. PMID:23246556

Hatchling turtles survive freezing during winter hibernation.

PubMed Central

Storey, K B; Storey, J M; Brooks, S P; Churchill, T A; Brooks, R J

1988-01-01

Hatchlings of the painted turtle (Chrysemys picta marginata) are unique as the only reptile and highest vertebrate life form known to tolerate the natural freezing of extracellular body fluids during winter hibernation. Turtles survived frequent exposures to temperatures as low as -6 degrees C to -8 degrees C in their shallow terrestrial nests over the 1987-1988 winter. Hatchlings collected in April 1988 had a mean supercooling point of -3.28 +/- 0.24 degrees C and survived 24 hr of freezing at -4 degrees C with 53.4% +/- 1.98% of total body water as ice. Recovery appeared complete after 20 hr of thawing at 3 degrees C. However, freezing at -10.9 degrees C, resulting in 67% ice, was lethal. A survey of possible cryoprotectants revealed a 2- to 3-fold increase in glucose content of liver and blood and a 3-fold increase in blood glycerol in response to freezing. Although quantitatively low, these responses by spring turtles strongly indicate that these may be the winter-active cryoprotectants. The total amino acid pool of blood also increased 2.25-fold in freezing-exposed turtles, and taurine accounted for 52% of the increase. Most organs accumulated high concentrations of lactate during freezing, a response to the ischemic state imposed by extracellular freezing. Changes in glycogen phosphorylase activity and levels of glucose 6-phosphate and fructose 2,6-bisphosphate were also consistent with a dependence on anaerobic glycolysis during freezing. Studies of the molecular mechanisms of natural freeze tolerance in these turtles may identify protective strategies that can be used in mammalian organ cryopreservation technology. PMID:3186730

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

PubMed

Duman, John G

2015-06-01

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.

Freeze-drying in novel container system: Characterization of heat and mass transfer in glass syringes.

PubMed

Patel, Sajal M; Pikal, Michael J

2010-07-01

This study is aimed at characterizing and understanding different modes of heat and mass transfer in glass syringes to develop a robust freeze-drying process. Two different holder systems were used to freeze-dry in syringes: an aluminum (Al) block and a plexiglass holder. The syringe heat transfer coefficient was characterized by a sublimation test using pure water. Mannitol and sucrose (5% w/v) were also freeze-dried, as model systems, in both the assemblies. Dry layer resistance was determined from manometric temperature measurement (MTM) and product temperature was measured using thermocouples, and was also determined from MTM. Further, freeze-drying process was also designed using Smart freeze-dryer to assess its application for freeze-drying in novel container systems. Heat and mass transfer in syringes were compared against the traditional container system (i.e., glass tubing vial). In the Al block, the heat transfer was via three modes: contact conduction, gas conduction, and radiation with gas conduction being the dominant mode of heat transfer. In the plexiglass holder, the heat transfer was mostly via radiation; convection was not involved. Also, MTM/Smart freeze-drying did work reasonably well for freeze-drying in syringes. When compared to tubing vials, product temperature decreases and hence drying time increases in syringes. (c) 2010 Wiley-Liss, Inc. and the American Pharmacists Association

Freeze-drying process monitoring using a cold plasma ionization device.

PubMed

Mayeresse, Y; Veillon, R; Sibille, P H; Nomine, C

2007-01-01

A cold plasma ionization device has been designed to monitor freeze-drying processes in situ by monitoring lyophilization chamber moisture content. This plasma device, which consists of a probe that can be mounted directly on the lyophilization chamber, depends upon the ionization of nitrogen and water molecules using a radiofrequency generator and spectrometric signal collection. The study performed on this probe shows that it is steam sterilizable, simple to integrate, reproducible, and sensitive. The limitations include suitable positioning in the lyophilization chamber, calibration, and signal integration. Sensitivity was evaluated in relation to the quantity of vials and the probe positioning, and correlation with existing methods, such as microbalance, was established. These tests verified signal reproducibility through three freeze-drying cycles. Scaling-up studies demonstrated a similar product signature for the same product using pilot-scale and larger-scale equipment. On an industrial scale, the method efficiently monitored the freeze-drying cycle, but in a larger industrial freeze-dryer the signal was slightly modified. This was mainly due to the positioning of the plasma device, in relation to the vapor flow pathway, which is not necessarily homogeneous within the freeze-drying chamber. The plasma tool is a relevant method for monitoring freeze-drying processes and may in the future allow the verification of current thermodynamic freeze-drying models. This plasma technique may ultimately represent a process analytical technology (PAT) approach for the freeze-drying process.

Detection of Collapse and Crystallization of Saccharide, Protein, and Mannitol Formulations by Optical Fibers in Lyophilization

PubMed Central

Horn, Jacqueline; Friess, Wolfgang

2018-01-01

The collapse temperature (Tc) and the glass transition temperature of freeze-concentrated solutions (Tg') as well as the crystallization behavior of excipients are important physicochemical characteristics which guide the cycle development in freeze-drying. The most frequently used methods to determine these values are differential scanning calorimetry (DSC) and freeze-drying microscopy (FDM). The objective of this study was to evaluate the optical fiber system (OFS) unit as alternative tool for the analysis of Tc, Tg' and crystallization events. The OFS unit was also tested as a potential online monitoring tool during freeze-drying. Freeze/thawing and freeze-drying experiments of sucrose, trehalose, stachyose, mannitol, and highly concentrated IgG1 and lysozyme solutions were carried out and monitored by the OFS. Comparative analyses were performed by DSC and FDM. OFS and FDM results correlated well. The crystallization behavior of mannitol could be monitored by the OFS during freeze/thawing as it can be done by DSC. Online monitoring of freeze-drying runs detected collapse of amorphous saccharide matrices. The OFS unit enabled the analysis of both Tc and crystallization processes, which is usually carried out by FDM and DSC. The OFS can hence be used as novel measuring device. Additionally, detection of these events during lyophilization facilitates online-monitoring. Thus the OFS is a new beneficial tool for the development and monitoring of freeze-drying processes. PMID:29435445

Evaluation of spin freezing versus conventional freezing as part of a continuous pharmaceutical freeze-drying concept for unit doses.

PubMed

De Meyer, L; Van Bockstal, P-J; Corver, J; Vervaet, C; Remon, J P; De Beer, T

2015-12-30

Spin-freezing as alternative freezing approach was evaluated as part of an innovative continuous pharmaceutical freeze-drying concept for unit doses. The aim of this paper was to compare the sublimation rate of spin-frozen vials versus traditionally frozen vials in a batch freeze-dryer, and its impact on total drying time. Five different formulations, each having a different dry cake resistance, were tested. After freezing, the traditionally frozen vials were placed on the shelves while the spin-frozen vials were placed in aluminum vial holders providing radial energy supply during drying. Different primary drying conditions and chamber pressures were evaluated. After 2h of primary drying, the amount of sublimed ice was determined in each vial. Each formulation was monitored in-line using NIR spectroscopy during drying to determine the sublimation endpoint and the influence of drying conditions upon total drying time. For all tested formulations and applied freeze-drying conditions, there was a significant higher sublimation rate in the spin-frozen vials. This can be explained by the larger product surface and the lower importance of product resistance because of the much thinner product layers in the spin frozen vials. The in-line NIR measurements allowed evaluating the influence of applied drying conditions on the drying trajectories. Copyright © 2015 Elsevier B.V. All rights reserved.

Inner ear tissue preservation by rapid freezing: improving fixation by high-pressure freezing and hybrid methods.

PubMed

Bullen, A; Taylor, R R; Kachar, B; Moores, C; Fleck, R A; Forge, A

2014-09-01

In the preservation of tissues in as 'close to life' state as possible, rapid freeze fixation has many benefits over conventional chemical fixation. One technique by which rapid freeze-fixation can be achieved, high pressure freezing (HPF), has been shown to enable ice crystal artefact-free freezing and tissue preservation to greater depths (more than 40 μm) than other quick-freezing methods. Despite increasingly becoming routine in electron microscopy, the use of HPF for the fixation of inner ear tissue has been limited. Assessment of the quality of preservation showed routine HPF techniques were suitable for preparation of inner ear tissues in a variety of species. Good preservation throughout the depth of sensory epithelia was achievable. Comparison to chemically fixed tissue indicated that fresh frozen preparations exhibited overall superior structural preservation of cells. However, HPF fixation caused characteristic artefacts in stereocilia that suggested poor quality freezing of the actin bundles. The hybrid technique of pre-fixation and high pressure freezing was shown to produce cellular preservation throughout the tissue, similar to that seen in HPF alone. Pre-fixation HPF produced consistent high quality preservation of stereociliary actin bundles. Optimising the preparation of samples with minimal artefact formation allows analysis of the links between ultrastructure and function in inner ear tissues. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

Detection of Collapse and Crystallization of Saccharide, Protein and Mannitol Formulations by Optical Fibers in Lyophilization

NASA Astrophysics Data System (ADS)

Horn, Jacqueline; Friess, Wolfgang

2018-01-01

The collapse temperature (Tc) and the glass transition temperature of freeze-concentrated solutions (Tg’) as well as the crystallization behavior of excipients are important physicochemical characteristics which guide the cycle development in freeze-drying. The most frequently used methods to determine these values are differential scanning calorimetry (DSC) and freeze-drying microscopy (FDM). The objective of this study was to evaluate the optical fiber system (OFS) unit as alternative tool for the analysis of Tc, Tg’ and crystallization events. The OFS unit was also tested as a potential online monitoring tool during freeze-drying. Freeze/thawing and freeze-drying experiments of sucrose, trehalose, stachyose, mannitol and highly concentrated IgG1 and lysozyme solutions were carried out and monitored by the OFS. Comparative analyses were performed by DSC and FDM. OFS and FDM results correlated well. The crystallization behavior of mannitol could be monitored by the OFS during freeze/thawing as it can be done by DSC. Online monitoring of freeze-drying runs detected collapse of amorphous saccharide matrices. The OFS unit enabled the analysis of both Tc and crystallization processes, which is usually carried out by FDM and DSC. The OFS can hence be used as novel measuring device. Additionally, detection of these events during lyophilization facilitate online-monitoring. Thus the OFS is a new beneficial tool for the development and monitoring of freeze-drying processes.

Development of a freeze-tolerant solar water heater using crosslinked polyethylene as a material of construction. Final report, June 18, 1976--October 1, 1977

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bradley, J.M.

The feasibility of building a freeze-tolerant absorber for a solar water heater out of carbon-black-reinforced crosslinked polyethylene has been explored. Ten-foot tube specimens made from various crosslinked polyethylene formulations were filled with water at various pressures, and then placed into a deep freeze, then thawed and frozen again for 100 freeze-thaw cycles, or until the tube specimen failed. Tube diameters were measured before and after each freezing to determine how much distention the freezing caused, and how much permanent distention was caused by the strains of repeated freezings. Five tube specimens containing water at as high as 80 psi survivedmore » 100 freeze-thaw cycles. Also, a flat plate collector was fabricated using as absorber surface a single 400 ft tube of carbon-black-reinforced crosslinked polyethylene in the form of a flat spiral coil and this collector was tested for performance at the Los Alamos Scientific Laboratory. The performance test indicates that the absorbancy of such a flat spiral coil to solar radiation is similar to typical black surfaces used on solar absorbers. Thus, it does seem very feasible that domestic water can be directly heated in a solar collector having an absorber made from crosslinked polyethylene, and that this collector can safely withstand at least 100 freeze-thaw cycles.« less

Repeated freezing induces a trade-off between cryoprotection and egg production in the goldenrod gall fly, Eurosta solidaginis.

PubMed

Marshall, Katie E; Sinclair, Brent J

2018-06-12

Internal ice formation leads to wholesale changes in ionic, osmotic and pH homeostasis, energy metabolism, and mechanical damage, across a small range of temperatures, and is thus an abiotic stressor that acts at a distinct, physiologically-relevant, threshold. Insects that experience repeated freeze-thaw cycles over winter will cross this stressor threshold many times over their lifespan. Here we examine the effect of repeatedly crossing the freezing threshold on short-term physiological parameters (metabolic reserves and cryoprotectant concentration) as well as long-term fitness-related performance (survival and egg production) in the freeze-tolerant goldenrod gall fly Eurosta solidaginis We exposed overwintering prepupae to a series of low temperatures (-10, -15, or -20 °C) with increasing numbers of freezing events (3, 6, or 10) with differing recovery periods between events (1, 5, or 10 days). Repeated freezing increased sorbitol concentration by about 50% relative to a single freezing episode, and prompted prepupae to modify long chain triacylglycerols to acetylated triacylglycerols. Long-term, repeated freezing did not significantly reduce survival, but did reduce egg production by 9.8% relative to a single freezing event. Exposure temperature did not affect any of these measures, suggesting that threshold crossing events may be more important to fitness than the intensity of stress in E. solidaginis overwintering. © 2018. Published by The Company of Biologists Ltd.

Freezing Bubbles

NASA Astrophysics Data System (ADS)

Kingett, Christian; Ahmadi, Farzad; Nath, Saurabh; Boreyko, Jonathan

2017-11-01

The two-stage freezing process of a liquid droplet on a substrate is well known; however, how bubbles freeze has not yet been studied. We first deposited bubbles on a silicon substrate that was chilled at temperatures ranging from -10 °C to -40 °C, while the air was at room temperature. We observed that the freeze front moved very slowly up the bubble, and in some cases, even came to a complete halt at a critical height. This slow freezing front propagation can be explained by the low thermal conductivity of the thin soap film, and can be observed more clearly when the bubble size or the surface temperature is increased. This delayed freezing allows the frozen portion of the bubble to cool the air within the bubble while the top part is still liquid, which induces a vapor pressure mismatch that either collapses the top or causes the top to pop. In cases where the freeze front reaches the top of the bubble, a portion of the top may melt and slowly refreeze; this can happen more than just once for a single bubble. We also investigated freezing bubbles inside of a freezer where the air was held at -20 °C. In this case, the bubbles freeze quickly and the ice grows radially from nucleation sites instead of perpendicular to the surface, which provides a clear contrast with the conduction limited room temperature bubbles.

Female Fertility Issues

Cancer.gov

Fertility issues are common in women getting cancer treatment. Fertility preservation options include egg freezing, embryo freezing, ovarian tissue freezing, ovarian shielding and ovarian transposition. Resources, support, and clinical trials are listed.

Non-Toxic, Low-Freezing, Drop-In Replacement Heat Transfer Fluids

NASA Technical Reports Server (NTRS)

Cutbirth, J. Michael

2012-01-01

A non-toxic, non-flammable, low-freezing heat transfer fluid is being developed for drop-in replacement within current and future heat transfer loops currently using water or alcohol-based coolants. Numerous water-soluble compounds were down-selected and screened for toxicological, physical, chemical, compatibility, thermodynamic, and heat transfer properties. Two fluids were developed, one with a freezing point near 0 C, and one with a suppressed freezing point. Both fluids contain an additive package to improve material compatibility and microbial resistance. The optimized sub-zero solution had a freezing point of 30 C, and a freezing volume expansion of 10-percent of water. The toxicity of the solutions was experimentally determined as LD(50) greater than 5g/kg. The solutions were found to produce minimal corrosion with materials identified by NASA as potentially existing in secondary cooling loops. Thermal/hydrodynamic performance exceeded that of glycol-based fluids with comparable freezing points for temperatures Tf greater than 20 C. The additive package was demonstrated as a buffering agent to compensate for CO2 absorption, and to prevent microbial growth. The optimized solutions were determined to have physically/chemically stable shelf lives for freeze/thaw cycles and longterm test loop tests.

The Structural Properties and Stability of Monoclonal Antibodies at Freezing Conditions

NASA Astrophysics Data System (ADS)

Perevozchikova, Tatiana; Zarraga, Isidro; Scherer, Thomas; Wagner, Norman; Liu, Yun

2013-03-01

Monoclonal Antibodies (MAb) have become a crucial therapeutic agent in a number of anti-cancer treatments. Due to the inherent unstable nature of proteins in an aqueous formulation, a freeze-drying method has been developed to maintain long-term stability of biotherapeutics. The microstructural changes in Mabs during freezing, however, remain not fully described, and it was proposed that the formed morphology of freeze drying samples could affect the final product quality after reconstitution. Furthermore, it is well known that proteins tend to aggregate during the freezing process if a careful processing procedure is not formulated. Small Angle Neutron Scattering (SANS) is a powerful tool to investigate the structural properties and interactions of Mabs during various stages of lyophilization in situ. Here we present the SANS results of freeze-thaw studies on two MAbs at several different freezing temperatures. While the chosen proteins share a significant sequence homology, their freezing properties are found to be strikingly distinctive. We also show the effect of excipients, concentration and quenching speed on the final morphology of the frozen samples. These findings provide critical information for more effective lyophilization schemes for therapeutic proteins, as well as increase our understanding on structural properties of proteins under cryogenic conditions.

Key composition optimization of meat processed protein source by vacuum freeze-drying technology.

PubMed

Ma, Yan; Wu, Xingzhuang; Zhang, Qi; Giovanni, Vigna; Meng, Xianjun

2018-05-01

Vacuum freeze-drying technology is a high technology content, a wide range of knowledge of technology in the field of drying technology is involved, it is also a method of the most complex drying equipment, the largest energy consumption, the highest cost of drying method, but due to the particularity of its dry goods: the freeze-drying food has the advantages of complex water performance is good, cooler and luster of freezing and drying food to maintain good products, less nutrient loss, light weight, easy to carry transportation, easy to long-term preservation, and on the quality is far superior to the obvious advantages of other dried food, making it become the forefront of drying technology research and development. The freeze-drying process of Chinese style ham and western Germany fruit tree tenderloin is studied in this paper, their eutectic point, melting point and collapse temperature, freeze-drying curve and its heat and mass transfer characteristics are got, then the precool temperature and the highest limiting temperature of sublimation interface are determined. The effect of system pressure on freeze-dried rate in freeze-drying process is discussed, and the method of regulating pressure circularly is determined.

Vernal freeze damage and genetic variation alter tree growth, chemistry, and insect interactions.

PubMed

Rubert-Nason, Kennedy F; Couture, John J; Gryzmala, Elizabeth A; Townsend, Philip A; Lindroth, Richard L

2017-11-01

Anticipated consequences of climate change in temperate regions include early spring warmup punctuated by intermittent hard freezes. Warm weather accelerates leaf flush in perennial woody species, potentially exposing vulnerable young tissues to damaging frosts. We employed a 2 × 6 randomized factorial design to examine how the interplay of vernal (springtime) freeze damage and genetic variation in a hardwood species (Populus tremuloides) influences tree growth, phytochemistry, and interactions with an insect herbivore (Chaitophorus stevensis). Acute effects of freezing included defoliation and mortality. Surviving trees exhibited reduced growth and altered biomass distribution. Reflushed leaves on these trees had lower mass per area, lower lignin concentrations, and higher nitrogen concentrations, altered chemical defence profiles, and supported faster aphid population growth. Many effects varied among plant genotypes and were related with herbivore performance. This study suggests that a single damaging vernal freeze event can alter tree-insect interactions through effects on plant growth and chemistry. Differential responses of various genotypes to freeze damage suggest that more frequent vernal freeze events could also influence natural selection, favouring trees with greater freeze hardiness, and more resistance or tolerance to herbivores following damage. © 2017 John Wiley & Sons Ltd.

Freeze-Thaw Durability of Air-Entrained Concrete

PubMed Central

Shang, Huai-Shuai; Yi, Ting-Hua

2013-01-01

One of the most damaging actions affecting concrete is the abrupt temperature change (freeze-thaw cycles). The types of deterioration of concrete structures by cyclic freeze-thaw can be largely classified into surface scaling (characterized by the weight loss) and internal crack growth (characterized by the loss of dynamic modulus of elasticity). The present study explored the durability of concrete made with air-entraining agent subjected to 0, 100, 200, 300, and 400 cycles of freeze-thaw. The experimental study of C20, C25, C30, C40, and C50 air-entrained concrete specimens was completed according to “the test method of long-term and durability on ordinary concrete” GB/T 50082-2009. The dynamic modulus of elasticity and weight loss of specimens were measured after different cycles of freeze-thaw. The influence of freeze-thaw cycles on the relative dynamic modulus of elasticity and weight loss was analyzed. The findings showed that the dynamic modulus of elasticity and weight decreased as the freeze-thaw cycles were repeated. They revealed that the C30, C40, and C50 air-entrained concrete was still durable after 300 cycles of freeze-thaw according to the experimental results. PMID:23576906

Freeze-thaw durability of air-entrained concrete.

PubMed

Shang, Huai-Shuai; Yi, Ting-Hua

2013-01-01

One of the most damaging actions affecting concrete is the abrupt temperature change (freeze-thaw cycles). The types of deterioration of concrete structures by cyclic freeze-thaw can be largely classified into surface scaling (characterized by the weight loss) and internal crack growth (characterized by the loss of dynamic modulus of elasticity). The present study explored the durability of concrete made with air-entraining agent subjected to 0, 100, 200, 300, and 400 cycles of freeze-thaw. The experimental study of C20, C25, C30, C40, and C50 air-entrained concrete specimens was completed according to "the test method of long-term and durability on ordinary concrete" GB/T 50082-2009. The dynamic modulus of elasticity and weight loss of specimens were measured after different cycles of freeze-thaw. The influence of freeze-thaw cycles on the relative dynamic modulus of elasticity and weight loss was analyzed. The findings showed that the dynamic modulus of elasticity and weight decreased as the freeze-thaw cycles were repeated. They revealed that the C30, C40, and C50 air-entrained concrete was still durable after 300 cycles of freeze-thaw according to the experimental results.

Mutual Influence of Mannitol and Trehalose on Crystallization Behavior in Frozen Solutions.

PubMed

Jena, Sampreeti; Suryanarayanan, Raj; Aksan, Alptekin

2016-06-01

Phase separation of trehalose during freeze-drying could render it ineffective as a lyoprotectant. The bulking agent, mannitol, on the other hand, should crystallize readily upon freezing. It is therefore imperative to understand the mutual interaction of these sugars during freezing to ensure preservation of the API during freeze-drying. We investigated the effect of mannitol to trehalose ratio (R) on the crystallization behavior of both solutes using Differential Scanning Calorimetry, X-Ray Crystallography and FTIR Spectrosopy during controlled freezing and annealing. When R = 1, crystallization of both mannitol (as hemihydrate) and trehalose (as α-anhydrate) were observed. When R ≥ 1, extent of mannitol crystallization was directly proportional to the value of R. When R < 1, trehalose completely suppressed mannitol crystallization. At R > 1, the freeze concentrate was heterogeneous and characterized by two glass transitions - the lower temperature transition (Tg") attributed to systems containing "extra" unfrozen water. When heated above Tg", crystallization of mannitol and the associated unfrozen water resulted in Tg' (glass transition temperature of the freeze-concentrate). R and not the total solute concentration, dictates the composition of the freeze concentrate as well as the physical stability of the excipients.

Freezing as an intervention to reduce the numbers of campylobacters isolated from chicken livers.

PubMed

Harrison, D; Corry, J E L; Tchórzewska, M A; Morris, V K; Hutchison, M L

2013-09-01

The aims of this study were (i) to determine the prevalence and numbers of campylobacters in 63 samples of raw livers purchased at retail across the UK and (ii) to investigate whether the freezing of chicken livers contaminated with Campylobacter was a reliable method for decontamination. Chicken livers naturally contaminated with campylobacters were subjected to freezing at -15 and -25°C for one day and 7 days. Numbers of campylobacters on the livers were determined immediately before and after a 24-h or 7-days freeze treatment and daily during 3 days post-thaw refrigerated storage. Freezing for 24 h at -25°C can reduce numbers of Campylobacter by up to 2 log10 CFU g(-1). Freezing the livers for 24 h at -25°C, thawing overnight in a fridge set to 4°C and refreezing for another 24 h at -25°C reduced the numbers of campylobacters by up to three logs. Reduction in the numbers of campylobacters was significantly greater following a second freeze treatment compared with a single freeze treatment. © 2013 Crown copyright. This article is published with the permission of the Controller of HMSO and the Queen's Printer for Scotland.

Gap-freezing approach for shortening the lyophilization cycle time of pharmaceutical formulations-demonstration of the concept.

PubMed

Kuu, Wei Y; Doty, Mark J; Rebbeck, Christine L; Hurst, William S; Cho, Yong K

2013-08-01

During gap freezing, vials are placed on a metal tray, which is separated from the shelf surface with a small air gap that eliminates significant conductive heat transfer from the shelf to the bottom of the vial. The purpose of this freezing approach is to reduce the lyophilization cycle time of various amorphous formulations by nearly isothermal freezing. Such isothermal freezing promotes the formation of large ice crystals, and thus large pores throughout the cake, which subsequently accelerates the primary drying rate. The nucleation temperature using gap freezing, for the experimental conditions tested, was in the range of -1°C to -6°C, much higher than the range of -10°C to -14°C found using conventional shelf freezing. Isothermal freezing becomes effective when the gap is greater than 3 mm. The pore sizes and cake resistance during primary drying for various formulations were determined using the pore diffusion model developed by Kuu et al. (Pharm Dev Technol, 2011, 16(4): 343-357). Reductions in primary drying time were 42% (for 10% sucrose), 45% (for 10% trehalose), and 33% (for 5% sucrose). Copyright © 2013 Wiley Periodicals, Inc.

Intracellular Freezing in the Infective Juveniles of Steinernema feltiae: An Entomopathogenic Nematode

PubMed Central

Ali, Farman; Wharton, David A.

2014-01-01

Taking advantage of their optical transparency, we clearly observed the third stage infective juveniles (IJs) of Steinernema feltiae freezing under a cryo-stage microscope. The IJs froze when the water surrounding them froze at −2°C and below. However, they avoid inoculative freezing at −1°C, suggesting cryoprotective dehydration. Freezing was evident as a sudden darkening and cessation of IJs' movement. Freeze substitution and transmission electron microscopy confirmed that the IJs of S. feltiae freeze intracellularly. Ice crystals were found in every compartment of the body. IJs frozen at high sub-zero temperatures (−1 and −3°C) survived and had small ice crystals. Those frozen at −10°C had large ice crystals and did not survive. However, the pattern of ice formation was not well-controlled and individual nematodes frozen at −3°C had both small and large ice crystals. IJs frozen by plunging directly into liquid nitrogen had small ice crystals, but did not survive. This study thus presents the evidence that S. feltiae is only the second freeze tolerant animal, after the Antarctic nematode Panagrolaimus davidi, shown to withstand extensive intracellular freezing. PMID:24769523

Freeze-cast alumina pore networks: Effects of freezing conditions and dispersion medium

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, S. M.; Xiao, X.; Faber, K. T.

Alumina ceramics were freeze-cast from water- and camphene-based slurries under varying freezing conditions and examined using X-ray computed tomography (XCT). Pore network characteristics, i.e., porosity, pore size, geometric surface area, and tortuosity, were measured from XCT reconstructions and the data were used to develop a model to predict feature size from processing conditions. Classical solidification theory was used to examine relationships between pore size, temperature gradients, and freezing front velocity. Freezing front velocity was subsequently predicted from casting conditions via the two-phase Stefan problem. Resulting models for water-based samples agreed with solidification-based theories predicting lamellar spacing of binary eutectic alloys,more » and models for camphene-based samples concurred with those for dendritic growth. Relationships between freezing conditions and geometric surface area were also modeled by considering the inverse relationship between pore size and surface area. Tortuosity was determined to be dependent primarily on the type of dispersion medium. (C) 2015 Elsevier Ltd. All rights reserved.« less

Effects of egg yolk, glycerol and the freezing rate on the viability and acrosomal structures of frozen ram spermatozoa.

PubMed

Watson, P F; Martin, I C

1975-04-01

The influence of egg yolk, glycerol and the freezing rate on the survival of ram spermatozoa and on the structure of their acrosomes after freezing was investigated. Egg yolk was shown to be beneficial not only during chilling but also during freezing; of the levels examined, 1-5% gave the greatest protection. Although the presence of glycerol in the diluent improved the survival of spermatozoa, increasing concentrations produced significant deterioration of the acrosomes. With closely controlled linear cooling rates, no overall difference was detected in the survival of spermatozoa frozen at rates between 6 and 24 degrees C per min. However, a significant interaction between freezing rate and the inclusion of glycerol in the diluent showed that glycerol was less important at the highest freezing rate. A sudden cooling phase near to the freezing point following the release of the latent heat of fusion was not detrimental to spermatozoa.

Pharmaceutical patent applications in freeze-drying.

PubMed

Ekenlebie, Edmond; Einfalt, Tomaž; Karytinos, Arianna Irò; Ingham, Andrew

2016-09-01

Injectable products are often the formulation of choice for new therapeutics; however, formulation in liquids often enhances degradation through hydrolysis. Thus, freeze-drying (lyophilization) is regularly used in pharmaceutical manufacture to reduce water activity. Here we examine its contribution to 'state of the art' and look at its future potential uses. A comprehensive search of patent databases was conducted to characterize the international patent landscape and trends in the use of freeze-drying. A total of 914 disclosures related to freeze-drying, lyophilization or drying of solid systems in pressures and temperatures equivalent to those of freeze-drying were considered over the period of 1992-2014. Current applications of sublimation technology were contrasted across two periods those with patents due to expire (1992-1993) and those currently filed. The number of freeze-drying technology patents has stabilized after initial activity across the biotechnology sector in 2011 and 2012. Alongside an increasing trend for patent submissions, freeze-drying submissions have slowed since 2002 and is indicative of a level of maturity.

Giant panda (Ailuropoda melanoleuca) sperm morphometry and function after repeated freezing and thawing.

PubMed

Santiago-Moreno, J; Esteso, M C; Pradiee, J; Castaño, C; Toledano-Díaz, A; O'Brien, E; Lopez-Sebastián, A; Martínez-Nevado, E; Delclaux, M; Fernández-Morán, J; Zhihe, Z

2016-05-01

This work examines the effects of subsequent cycles of freezing-thawing on giant panda (Ailuropoda melanoleuca) sperm morphometry and function, and assesses whether density-gradient centrifugation (DGC) can increase the number of freezing-thawing cycles this sperm can withstand. A sperm sample was collected by electroejaculation from a mature giant panda and subjected to five freezing-thawing cycles. Although repeated freezing-thawing negatively affected (P < 0.05) sperm motility and membrane integrity, in both nonselected and DCG-selected sperm samples, >60% of the sperm cells in both treatments showed acrosome integrity even after the fifth freezing cycle. In fresh semen, the sperm head length was 4.7 μm, the head width 3.6 μm, area 14.3 μm(2) and perimeter length 14.1 μm. The present results suggest that giant panda sperm trends to be resistant to repeated freezing-thawing, even without DGC selection. © 2015 Blackwell Verlag GmbH.

Ultrasound assisted immersion freezing of broccoli (Brassica oleracea L. var. botrytis L.).

PubMed

Xin, Ying; Zhang, Min; Adhikari, Benu

2014-09-01

The aim of this study was to research the ultrasound-assisted freezing (UAF) of broccoli. CaCl2 solution was used as freezing medium. The comparative advantage of using UAF over normal freezing on the freezing time, cell-wall bound calcium to total calcium ratio, textural properties, color, drip loss and L-ascorbic acid contents was evaluated. The application of UAF at selected acoustic intensity with a range of 0.250-0.412 W/cm(2) decreased the freezing time and the loss of cell-wall bound calcium content. Compared to normal freezing, the values of textural properties, color, L-ascorbic acid content were better preserved and the drip loss was significantly minimized by the application of UAF. However, when outside that range of acoustic intensity, the quality of the ultrasound-assisted frozen broccoli was inferior compared to that of the normally frozen samples. Selected the appropriate acoustic intensity was very important for the application of UAF. Copyright © 2014 Elsevier B.V. All rights reserved.

Recent developments in novel freezing and thawing technologies applied to foods.

PubMed

Wu, Xiao-Fei; Zhang, Min; Adhikari, Benu; Sun, Jincai

2017-11-22

This article reviews the recent developments in novel freezing and thawing technologies applied to foods. These novel technologies improve the quality of frozen and thawed foods and are energy efficient. The novel technologies applied to freezing include pulsed electric field pre-treatment, ultra-low temperature, ultra-rapid freezing, ultra-high pressure and ultrasound. The novel technologies applied to thawing include ultra-high pressure, ultrasound, high voltage electrostatic field (HVEF), and radio frequency. Ultra-low temperature and ultra-rapid freezing promote the formation and uniform distribution of small ice crystals throughout frozen foods. Ultra-high pressure and ultrasound assisted freezing are non-thermal methods and shorten the freezing time and improve product quality. Ultra-high pressure and HVEF thawing generate high heat transfer rates and accelerate the thawing process. Ultrasound and radio frequency thawing can facilitate thawing process by volumetrically generating heat within frozen foods. It is anticipated that these novel technologies will be increasingly used in food industries in the future.

The Bergschrund Hypothesis Revisited

NASA Astrophysics Data System (ADS)

Sanders, J. W.; Cuffey, K. M.; MacGregor, K. R.

2009-12-01

After Willard Johnson descended into the Lyell Glacier bergschrund nearly 140 years ago, he proposed that the presence of the bergschrund modulated daily air temperature fluctuations and enhanced freeze-thaw processes. He posited that glaciers, through their ability to birth bergschrunds, are thus able to induce rapid cirque headwall retreat. In subsequent years, many researchers challenged the bergschrund hypothesis on grounds that freeze-thaw events did not occur at depth in bergschrunds. We propose a modified version of Johnson’s original hypothesis: that bergschrunds maintain subfreezing temperatures at values that encourage rock fracture via ice lensing because they act as a cold air trap in areas that would otherwise be held near zero by temperate glacial ice. In support of this claim we investigated three sections of the bergschrund at the West Washmawapta Glacier, British Columbia, Canada, which sits in an east-facing cirque. During our bergschrund reconnaissance we installed temperature sensors at multiple elevations, light sensors at depth in 2 of the 3 locations and painted two 1 m2 sections of the headwall. We first emphasize bergschrunds are not wanting for ice: verglas covers significant fractions of the headwall and icicles dangle from the base of bödens or overhanging rocks. If temperature, rather than water availability, is the limiting factor governing ice-lensing rates, our temperature records demonstrate that the bergschrund provides a suitable environment for considerable rock fracture. At the three sites (north, west, and south walls), the average temperature at depth from 9/3/2006 to 8/6/2007 was -3.6, -3.6, and -2.0 °C, respectively. During spring, when we observed vast amounts of snow melt trickle in to the bergschrund, temperatures averaged -3.7, -3.8, and -2.2 °C, respectively. Winter temperatures are even lower: -8.5, -7.3, and -2.4 °C, respectively. Values during the following year were similar. During the fall, diurnal temperature variation at depth is recognizable but stabilizes as light intensity falls to zero. Nevertheless, large amplitude dips in temperature are still noticeable at all elevations. Diurnal temperature swings do not reappear until July. Within the painted rock sections, only 1 x10-3 m3 of rock completely detached from the wall. In light of our observations and temperature records, we believe the bergschrund does play a major role in cirque formation. However, further efforts to record environmental variables necessary for ice lensing at the heads of glaciers are warranted, especially at locations below the bergschrund.

Self-assembled multicompartment liquid crystalline lipid carriers for protein, peptide, and nucleic acid drug delivery.

PubMed

Angelova, Angelina; Angelov, Borislav; Mutafchieva, Rada; Lesieur, Sylviane; Couvreur, Patrick

2011-02-15

Lipids and lipopolymers self-assembled into biocompatible nano- and mesostructured functional materials offer many potential applications in medicine and diagnostics. In this Account, we demonstrate how high-resolution structural investigations of bicontinuous cubic templates made from lyotropic thermosensitive liquid-crystalline (LC) materials have initiated the development of innovative lipidopolymeric self-assembled nanocarriers. Such structures have tunable nanochannel sizes, morphologies, and hierarchical inner organizations and provide potential vehicles for the predictable loading and release of therapeutic proteins, peptides, or nucleic acids. This Account shows that structural studies of swelling of bicontinuous cubic lipid/water phases are essential for overcoming the nanoscale constraints for encapsulation of large therapeutic molecules in multicompartment lipid carriers. For the systems described here, we have employed time-resolved small-angle X-ray scattering (SAXS) and high-resolution freeze-fracture electronic microscopy (FF-EM) to study the morphology and the dynamic topological transitions of these nanostructured multicomponent amphiphilic assemblies. Quasi-elastic light scattering and circular dichroism spectroscopy can provide additional information at the nanoscale about the behavior of lipid/protein self-assemblies under conditions that approximate physiological hydration. We wanted to generalize these findings to control the stability and the hydration of the water nanochannels in liquid-crystalline lipid nanovehicles and confine therapeutic biomolecules within these structures. Therefore we analyzed the influence of amphiphilic and soluble additives (e.g. poly(ethylene glycol)monooleate (MO-PEG), octyl glucoside (OG), proteins) on the nanochannels' size in a diamond (D)-type bicontinuous cubic phase of the lipid glycerol monooleate (MO). At body temperature, we can stabilize long-living swollen states, corresponding to a diamond cubic phase with large water channels. Time-resolved X-ray diffraction (XRD) scans allowed us to detect metastable intermediate and coexisting structures and monitor the temperature-induced phase sequences of mixed systems containing glycerol monooleate, a soluble protein macromolecule, and an interfacial curvature modulating agent. These observed states correspond to the stages of the growth of the nanofluidic channel network. With the application of a thermal stimulus, the system becomes progressively more ordered into a double-diamond cubic lattice formed by a bicontinuous lipid membrane. High-resolution freeze-fracture electronic microscopy indicates that nanodomains are induced by the inclusion of proteins into nanopockets of the supramolecular cubosomic assemblies. These results contribute to the understanding of the structure and dynamics of functionalized self-assembled lipid nanosystems during stimuli-triggered LC phase transformations.

A molecular dynamics study of freezing in a confined geometry

NASA Technical Reports Server (NTRS)

Ma, Wen-Jong; Banavar, Jayanth R.; Koplik, Joel

1992-01-01

The dynamics of freezing of a Lennard-Jones liquid in narrow channels bounded by molecular walls is studied by computer simulation. The time development of ordering is quantified and a novel freezing mechanism is observed. The liquid forms layers and subsequent in-plane ordering within a layer is accompanied by a sharpening of the layer in the transverse direction. The effects of channel size, the methods of quench, the liquid-wall interaction and the roughness of walls on the freezing mechanism are elucidated. Comparison with recent experiments on freezing in confined geometries is presented.

Hidden transition in multiferroic and magnetodielectric CuCrO2 evidenced by ac-susceptibility

NASA Astrophysics Data System (ADS)

Shukla, Kaushak K.; Pal, Arkadeb; Singh, Abhishek; Singh, Rahul; Saha, J.; Sinha, A. K.; Ghosh, A. K.; Patnaik, S.; Awasthi, A. M.; Chatterjee, Sandip

2017-04-01

Ferroelectric polarization, magnetic-field dependence of the dielectric constant and ac and dc magnetizations of frustrated CuCrO2 have been measured. A new spin freezing transition below 32 K is observed which is thermally driven. The nature of the spin freezing is to be a single-ion process. Dilution by the replacements of Cr ions by magnetic Mn ions showed suppression of the spin freezing transition suggesting it to be fundamentally a single-ion freezing process. The observed freezing, which is seemingly associated to geometrical spin frustration, represents a novel form of magnetic glassy behavior.

Design of a blood-freezing system for leukemia research

NASA Technical Reports Server (NTRS)

Williams, T. E.; Cygnarowicz, T. A.

1978-01-01

Leukemia research involves the use of cryogenic freezing and storage equipment. In a program being carried out at the National Cancer Institute (NCI), bone marrow (white blood cells) was frozen using a standard cryogenic biological freezer. With this system, it is difficult to maintain the desired rate of freezing and repeatability from sample to sample. A freezing system was developed that satisfies the requirements for a repeatable, constant freezing rate. The system was delivered to NIC and is now operational. This report describes the design of the major subsystems, the analyses, the operating procedure, and final system test results.

Effect of particle surface area on ice active site densities retrieved from droplet freezing spectra

NASA Astrophysics Data System (ADS)

Beydoun, Hassan; Polen, Michael; Sullivan, Ryan C.

2016-10-01

Heterogeneous ice nucleation remains one of the outstanding problems in cloud physics and atmospheric science. Experimental challenges in properly simulating particle-induced freezing processes under atmospherically relevant conditions have largely contributed to the absence of a well-established parameterization of immersion freezing properties. Here, we formulate an ice active, surface-site-based stochastic model of heterogeneous freezing with the unique feature of invoking a continuum assumption on the ice nucleating activity (contact angle) of an aerosol particle's surface that requires no assumptions about the size or number of active sites. The result is a particle-specific property g that defines a distribution of local ice nucleation rates. Upon integration, this yields a full freezing probability function for an ice nucleating particle. Current cold plate droplet freezing measurements provide a valuable and inexpensive resource for studying the freezing properties of many atmospheric aerosol systems. We apply our g framework to explain the observed dependence of the freezing temperature of droplets in a cold plate on the concentration of the particle species investigated. Normalizing to the total particle mass or surface area present to derive the commonly used ice nuclei active surface (INAS) density (ns) often cannot account for the effects of particle concentration, yet concentration is typically varied to span a wider measurable freezing temperature range. A method based on determining what is denoted an ice nucleating species' specific critical surface area is presented and explains the concentration dependence as a result of increasing the variability in ice nucleating active sites between droplets. By applying this method to experimental droplet freezing data from four different systems, we demonstrate its ability to interpret immersion freezing temperature spectra of droplets containing variable particle concentrations. It is shown that general active site density functions, such as the popular ns parameterization, cannot be reliably extrapolated below this critical surface area threshold to describe freezing curves for lower particle surface area concentrations. Freezing curves obtained below this threshold translate to higher ns values, while the ns values are essentially the same from curves obtained above the critical area threshold; ns should remain the same for a system as concentration is varied. However, we can successfully predict the lower concentration freezing curves, which are more atmospherically relevant, through a process of random sampling from g distributions obtained from high particle concentration data. Our analysis is applied to cold plate freezing measurements of droplets containing variable concentrations of particles from NX illite minerals, MCC cellulose, and commercial Snomax bacterial particles. Parameterizations that can predict the temporal evolution of the frozen fraction of cloud droplets in larger atmospheric models are also derived from this new framework.

Freeze injury to southern pine seedlings

Treesearch

David B. South

2006-01-01

Freeze injury to roots and shoots of pines is affected by genotype and nursery practices. Local sources of shortleaf pine and Virginia pine that are grown in nurseries in USDA hardiness Zones 6 and 7a are relatively freeze tolerant. However, loblolly pine, slash pine, and longleaf pine seedlings have been injured by a number of freeze events (0 to 24 °F) in hardiness...

Asymmetric pedunculopontine network connectivity in parkinsonian patients with freezing of gait

PubMed Central

Fling, Brett W.; Cohen, Rajal G.; Mancini, Martina; Nutt, John G.; Fair, Damian A.

2013-01-01

Freezing of gait is one of the most debilitating symptoms in Parkinson’s disease as it causes falls and reduces mobility and quality of life. The pedunculopontine nucleus is one of the major nuclei of the mesencephalic locomotor region and has neurons related to anticipatory postural adjustments preceding step initiation as well as to the step itself, thus it may be critical for coupling posture and gait to avoid freezing. Because freezing of gait and postural impairments have been related to frontal lesions and frontal dysfunction such as executive function, we hypothesized that freezing is associated with disrupted connectivity between midbrain locomotor regions and medial frontal cortex. We used diffusion tensor imaging to quantify structural connectivity of the pedunculopontine nucleus in patients with Parkinson’s disease with freezing of gait, without freezing, and healthy age-matched controls. We also included behavioural tasks to gauge severity of freezing of gait, quantify gait metrics, and assess executive cognitive functions to determine whether between-group differences in executive dysfunction were related to pedunculopontine nucleus structural network connectivity. Using seed regions from the pedunculopontine nucleus, we were able to delineate white matter connections between the spinal cord, cerebellum, pedunculopontine nucleus, subcortical and frontal/prefrontal cortical regions. The current study is the first to demonstrate differences in structural connectivity of the identified locomotor pathway in patients with freezing of gait. We report reduced connectivity of the pedunculopontine nucleus with the cerebellum, thalamus and multiple regions of the frontal cortex. Moreover, these structural differences were observed solely in the right hemisphere of patients with freezing of gait. Finally, we show that the more left hemisphere-lateralized the pedunculopontine nucleus tract volume, the poorer the performance on cognitive tasks requiring the initiation of appropriate actions and/or the inhibition of inappropriate actions, specifically within patients with freezing. These results support the notion that freezing of gait is strongly related to structural deficits in the right hemisphere’s locomotor network involving prefrontal cortical areas involved in executive inhibition function. PMID:23824487

Freezing and fractionation: effects of preservation on carbon and nitrogen stable isotope ratios of some limnetic organisms.

PubMed

Wolf, J Marshall; Johnson, Brett; Silver, Douglas; Pate, William; Christianson, Kyle

2016-03-15

Stable isotopes of carbon and nitrogen have become important natural tracers for studying food-web structure and function. Considerable research has demonstrated that chemical preservatives and fixatives shift the isotopic ratios of aquatic organisms. Much less is known about the effects of freezing as a preservation method although this technique is commonly used. We conducted a controlled experiment to test the effects of freezing (-10 °C) and flash freezing (–79 °C) on the carbon and nitrogen isotope ratios of zooplankton (Cladocera), Mysis diluviana and Rainbow Trout (Oncorhynchus mykiss). Subsamples (~0.5 mg) of dried material were analyzed for percentage carbon, percentage nitrogen, and the relative abundance of stable carbon and nitrogen isotopes (δ13C and δ15N values) using a Carlo Erba NC2500 elemental analyzer interfaced to a ThermoFinnigan MAT Delta Plus isotope ratio mass spectrometer. The effects of freezing were taxon-dependent. Freezing had no effect on the isotopic or elemental values of Rainbow Trout muscle. Effects on the δ13C and δ15N values of zooplankton and Mysis were statistically significant but small relative to typical values of trophic fractionation. The treatment-control offsets had larger absolute values for Mysis (δ13C: ≤0.76 ± 0.41‰, δ15N: ≤0.37 ± 0.16‰) than for zooplankton (δ13C: ≤0.12 ± 0.06‰, δ15N: ≤0.30 ± 0.27‰). The effects of freezing were more variable for the δ13C values of Mysis, and more variable for the δ15N values of zooplankton. Generally, both freezing methods reduced the carbon content of zooplankton and Mysis, but freezing had a negative effect on the %N of zooplankton and a positive effect on the %N of Mysis. The species-dependencies and variability of freezing effects on aquatic organisms suggest that more research is needed to understand the mechanisms responsible for freezing-related fractionation before standardized protocols for freezing as a preservation method can be adopted.

Implementation of a process analytical technology system in a freeze-drying process using Raman spectroscopy for in-line process monitoring.

PubMed

De Beer, T R M; Allesø, M; Goethals, F; Coppens, A; Heyden, Y Vander; De Diego, H Lopez; Rantanen, J; Verpoort, F; Vervaet, C; Remon, J P; Baeyens, W R G

2007-11-01

The aim of the present study was to propose a strategy for the implementation of a Process Analytical Technology system in freeze-drying processes. Mannitol solutions, some of them supplied with NaCl, were used as models to freeze-dry. Noninvasive and in-line Raman measurements were continuously performed during lyophilization of the solutions to monitor real time the mannitol solid state, the end points of the different process steps (freezing, primary drying, secondary drying), and physical phenomena occurring during the process. At-line near-infrared (NIR) and X-ray powder diffractometry (XRPD) measurements were done to confirm the Raman conclusions and to find out additional information. The collected spectra during the processes were analyzed using principal component analysis and multivariate curve resolution. A two-level full factorial design was used to study the significant influence of process (freezing rate) and formulation variables (concentration of mannitol, concentration of NaCl, volume of freeze-dried sample) upon freeze-drying. Raman spectroscopy was able to monitor (i) the mannitol solid state (amorphous, alpha, beta, delta, and hemihydrate), (ii) several process step end points (end of mannitol crystallization during freezing, primary drying), and (iii) physical phenomena occurring during freeze-drying (onset of ice nucleation, onset of mannitol crystallization during the freezing step, onset of ice sublimation). NIR proved to be a more sensitive tool to monitor sublimation than Raman spectroscopy, while XRPD helped to unravel the mannitol hemihydrate in the samples. The experimental design results showed that several process and formulation variables significantly influence different aspects of lyophilization and that both are interrelated. Raman spectroscopy (in-line) and NIR spectroscopy and XRPD (at-line) not only allowed the real-time monitoring of mannitol freeze-drying processes but also helped (in combination with experimental design) us to understand the process.

Anhydrobiosis and freezing-tolerance: adaptations that facilitate the establishment of Panagrolaimus nematodes in polar habitats.

PubMed

McGill, Lorraine M; Shannon, Adam J; Pisani, Davide; Félix, Marie-Anne; Ramløv, Hans; Dix, Ilona; Wharton, David A; Burnell, Ann M

2015-01-01

Anhydrobiotic animals can survive the loss of both free and bound water from their cells. While in this state they are also resistant to freezing. This physiology adapts anhydrobiotes to harsh environments and it aids their dispersal. Panagrolaimus davidi, a bacterial feeding anhydrobiotic nematode isolated from Ross Island Antarctica, can survive intracellular ice formation when fully hydrated. A capacity to survive freezing while fully hydrated has also been observed in some other Antarctic nematodes. We experimentally determined the anhydrobiotic and freezing-tolerance phenotypes of 24 Panagrolaimus strains from tropical, temperate, continental and polar habitats and we analysed their phylogenetic relationships. We found that several other Panagrolaimus isolates can also survive freezing when fully hydrated and that tissue extracts from these freezing-tolerant nematodes can inhibit the growth of ice crystals. We show that P. davidi belongs to a clade of anhydrobiotic and freezing-tolerant panagrolaimids containing strains from temperate and continental regions and that P. superbus, an early colonizer at Surtsey island, Iceland after its volcanic formation, is closely related to a species from Pennsylvania, USA. Ancestral state reconstructions show that anhydrobiosis evolved deep in the phylogeny of Panagrolaimus. The early-diverging Panagrolaimus lineages are strongly anhydrobiotic but weakly freezing-tolerant, suggesting that freezing tolerance is most likely a derived trait. The common ancestors of the davidi and the superbus clades were anhydrobiotic and also possessed robust freezing tolerance, along with a capacity to inhibit the growth and recrystallization of ice crystals. Unlike other endemic Antarctic nematodes, the life history traits of P. davidi do not show evidence of an evolved response to polar conditions. Thus we suggest that the colonization of Antarctica by P. davidi and of Surtsey by P. superbus may be examples of recent "ecological fitting" of freezing-tolerant anhydrobiotic propagules to the respective abiotic conditions in Ross Island and Surtsey.

Bursts of CO2 released during freezing offer a new perspective on avoidance of winter embolism in trees.

PubMed

Lintunen, A; Lindfors, L; Kolari, P; Juurola, E; Nikinmaa, E; Hölttä, T

2014-12-01

Woody plants can suffer from winter embolism as gas bubbles are formed in the water-conducting conduits when freezing occurs: gases are not soluble in ice, and the bubbles may expand and fill the conduits with air during thawing. A major assumption usually made in studies of winter embolism formation is that all of the gas dissolved in the xylem sap is trapped within the conduits and forms bubbles during freezing. The current study tested whether this assumption is actually valid, or whether efflux of gases from the stem during freezing reduces the occurrence of embolism. CO2 efflux measurements were conducted during freezing experiments for saplings of three Scots pine (Pinus sylvestris) and three Norway spruce (Picea abies) trees under laboratory conditions, and the magnitudes of the freezing-related bursts of CO2 released from the stems were analysed using a previously published mechanistic model of CO2 production, storage, diffusion and efflux from a tree stem. The freezing-related bursts of CO2 released from a mature Scots pine tree growing in field conditions were also measured and analysed. Substantial freezing-related bursts of CO2 released from the stem were found to occur during both the laboratory experiments and under field conditions. In the laboratory, the fraction of CO2 released from the stem ranged between 27 and 96 % of the total CO2 content within the stem. All gases dissolved in the xylem sap are not trapped within the ice in the stem during freezing, as has previously been assumed, thus adding a new dimension to the understanding of winter embolism formation. The conduit water volume not only determines the volume of bubbles formed during freezing, but also the efficiency of gas efflux out of the conduit during the freezing process. © The Author 2014. Published by Oxford University Press on behalf of the Annals of Botany Company.

The Role of Hydrogen Peroxide in Mediating the Mechanical Wounding-Induced Freezing Tolerance in Wheat

PubMed Central

Si, Tong; Wang, Xiao; Zhao, Chunzhao; Huang, Mei; Cai, Jian; Zhou, Qin; Dai, Tingbo; Jiang, Dong

2018-01-01

Systemic wound response (SWR), a well-characterized systemic signaling response, plays crucial roles in plant defense responses. Progress in understanding of the SWR in abiotic stress has also been aided by the researchers. However, the function of SWR in freezing stress remains elusive. In this study, we showed that local mild mechanical wounding enhanced freezing tolerance in newly occurred systemic leaves of wheat plants (Triticum aestivum L.). Wounding significantly increased the maximal photochemical efficiency of photosystem II, net photosynthetic rate, and the activities of the antioxidant enzymes under freezing stress. Wounding also alleviated freezing-induced chlorophyll decomposition, electrolyte leakage, water lose, and membrane peroxidation. In addition, wounding-induced freezing stress mitigation was closely associated with the ratio between reduced glutathione (GSH) and oxidized glutathione (GSSG), and the ratio between ascorbate (AsA) and dehydroascorbate (DHA), as well as the contents of total soluble sugars and free amino acids. Importantly, pharmacological study showed that wounding-induced freezing tolerance was substantially arrested by pretreatment of wheat leaves with the scavenger of hydrogen peroxide (H2O2) or the inhibitor of NADPH oxidase (RBOH). These results support the hypothesis that local mechanical wounding-induced SWR in newly occurred leaves is largely attributed to RBOH-dependent H2O2 production, which may subsequently induce freezing tolerance in wheat plants. This mechanism may have a potential application to reduce the yield losses of wheat under late spring freezing conditions. Highlights: In our previous research, we found that local mechanical wounding could induce freezing tolerance in the upper systemic leaves of wheat plants. Surprisingly, in this paper, we further demonstrated that local mechanical wounding could also increase freezing resistance in newly occurred leaves of wheat plants. RBOH mediated H2O2 and ascorbate–glutathione cycle participate in this systemic wound response. PMID:29593774

Anhydrobiosis and Freezing-Tolerance: Adaptations That Facilitate the Establishment of Panagrolaimus Nematodes in Polar Habitats

PubMed Central

McGill, Lorraine M.; Shannon, Adam J.; Pisani, Davide; Félix, Marie-Anne; Ramløv, Hans; Dix, Ilona; Wharton, David A.; Burnell, Ann M.

2015-01-01

Anhydrobiotic animals can survive the loss of both free and bound water from their cells. While in this state they are also resistant to freezing. This physiology adapts anhydrobiotes to harsh environments and it aids their dispersal. Panagrolaimus davidi, a bacterial feeding anhydrobiotic nematode isolated from Ross Island Antarctica, can survive intracellular ice formation when fully hydrated. A capacity to survive freezing while fully hydrated has also been observed in some other Antarctic nematodes. We experimentally determined the anhydrobiotic and freezing-tolerance phenotypes of 24 Panagrolaimus strains from tropical, temperate, continental and polar habitats and we analysed their phylogenetic relationships. We found that several other Panagrolaimus isolates can also survive freezing when fully hydrated and that tissue extracts from these freezing-tolerant nematodes can inhibit the growth of ice crystals. We show that P. davidi belongs to a clade of anhydrobiotic and freezing-tolerant panagrolaimids containing strains from temperate and continental regions and that P. superbus, an early colonizer at Surtsey island, Iceland after its volcanic formation, is closely related to a species from Pennsylvania, USA. Ancestral state reconstructions show that anhydrobiosis evolved deep in the phylogeny of Panagrolaimus. The early-diverging Panagrolaimus lineages are strongly anhydrobiotic but weakly freezing-tolerant, suggesting that freezing tolerance is most likely a derived trait. The common ancestors of the davidi and the superbus clades were anhydrobiotic and also possessed robust freezing tolerance, along with a capacity to inhibit the growth and recrystallization of ice crystals. Unlike other endemic Antarctic nematodes, the life history traits of P. davidi do not show evidence of an evolved response to polar conditions. Thus we suggest that the colonization of Antarctica by P. davidi and of Surtsey by P. superbus may be examples of recent “ecological fitting” of freezing-tolerant anhydrobiotic propagules to the respective abiotic conditions in Ross Island and Surtsey. PMID:25747673

A low membrane lipid phase transition temperature is associated with a high cryotolerance of Lactobacillus delbrueckii subspecies bulgaricus CFL1.

PubMed

Gautier, J; Passot, S; Pénicaud, C; Guillemin, H; Cenard, S; Lieben, P; Fonseca, F

2013-09-01

The mechanisms of cellular damage that lactic acid bacteria incur during freeze-thaw processes have not been elucidated to date. Fourier transform infrared spectroscopy was used to investigate in situ the lipid phase transition behavior of the membrane of Lactobacillus delbrueckii ssp. bulgaricus CFL1 cells during the freeze-thaw process. Our objective was to relate the lipid membrane behavior to membrane integrity losses during freezing and to cell-freezing resistance. Cells were produced by using 2 different culture media: de Man, Rogosa, and Sharpe (MRS) broth (complex medium) or mild whey-based medium (minimal medium commonly used in the dairy industry), to obtain different membrane lipid compositions corresponding to different recovery rates of cell viability and functionality after freezing. The lipid membrane behavior studied by Fourier transform infrared spectroscopy was found to be different according to the cell lipid composition and cryotolerance. Freeze-resistant cells, exhibiting a higher content of unsaturated and cyclic fatty acids, presented a lower lipid phase transition temperature (Ts) during freezing (Ts=-8°C), occurring within the same temperature range as the ice nucleation, than freeze-sensitive cells (Ts=+22°C). A subzero value of lipid phase transition allowed the maintenance of the cell membrane in a relatively fluid state during freezing, thus facilitating water flux from the cell and the concomitant volume reduction following ice formation in the extracellular medium. In addition, the lipid phase transition of freeze-resistant cells occurred within a short temperature range, which could be ascribed to a reduced number of fatty acids, representing more than 80% of the total. This short lipid phase transition could be associated with a limited phenomenon of lateral phase separation and membrane permeabilization. This work highlights that membrane phase transitions occurring during freeze-thawing play a fundamental role in the cryotolerance of Lb. delbrueckii ssp. bulgaricus CFL1 cells. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Air-Cooled Stack Freeze Tolerance Freeze Failure Modes and Freeze Tolerance Strategies for GenDriveTM Material Handling Application Systems and Stacks Final Scientific Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hancock, David, W.

2012-02-14

Air-cooled stack technology offers the potential for a simpler system architecture (versus liquid-cooled) for applications below 4 kilowatts. The combined cooling and cathode air allows for a reduction in part count and hence a lower cost solution. However, efficient heat rejection challenges escalate as power and ambient temperature increase. For applications in ambient temperatures below freezing, the air-cooled approach has additional challenges associated with not overcooling the fuel cell stack. The focus of this project was freeze tolerance while maintaining all other stack and system requirements. Through this project, Plug Power advanced the state of the art in technology formore » air-cooled PEM fuel cell stacks and related GenDrive material handling application fuel cell systems. This was accomplished through a collaborative work plan to improve freeze tolerance and mitigate freeze-thaw effect failure modes within innovative material handling equipment fuel cell systems designed for use in freezer forklift applications. Freeze tolerance remains an area where additional research and understanding can help fuel cells to become commercially viable. This project evaluated both stack level and system level solutions to improve fuel cell stack freeze tolerance. At this time, the most cost effective solutions are at the system level. The freeze mitigation strategies developed over the course of this project could be used to drive fuel cell commercialization. The fuel cell system studied in this project was Plug Power's commercially available GenDrive platform providing battery replacement for equipment in the material handling industry. The fuel cell stacks were Ballard's commercially available FCvelocity 9SSL (9SSL) liquid-cooled PEM fuel cell stack and FCvelocity 1020ACS (Mk1020) air-cooled PEM fuel cell stack.« less

Bulk water freezing dynamics on superhydrophobic surfaces

NASA Astrophysics Data System (ADS)

Chavan, S.; Carpenter, J.; Nallapaneni, M.; Chen, J. Y.; Miljkovic, N.

2017-01-01

In this study, we elucidate the mechanisms governing the heat-transfer mediated, non-thermodynamic limited, freezing delay on non-wetting surfaces for a variety of characteristic length scales, Lc (volume/surface area, 3 mm < Lc < 6 mm) using carefully designed freezing experiments in a temperature-controlled, zero-humidity environment on thin water slabs. To probe the effect of surface wettability, we investigated the total time for room temperature water to completely freeze into ice on superhydrophilic ( θaapp→ 0°), hydrophilic (0° < θa < 90°), hydrophobic (90° < θa < 125°), and superhydrophobic ( θaapp→ 180°) surfaces. Our results show that at macroscopic length scales, heat conduction through the bulk water/ice layer dominates the freezing process when compared to heat conduction through the functional coatings or nanoscale gaps at the superhydrophobic substrate-water/ice interface. In order to verify our findings, and to determine when the surface structure thermal resistance approaches the water/ice resistance, we fabricated and tested the additional substrates coated with commercial superhydrophobic spray coatings, showing a monotonic increase in freezing time with coating thickness. The added thermal resistance of thicker coatings was much larger than that of the nanoscale superhydrophobic features, which reduced the droplet heat transfer and increased the total freezing time. Transient finite element method heat transfer simulations of the water slab freezing process were performed to calculate the overall heat transfer coefficient at the substrate-water/ice interface during freezing, and shown to be in the range of 1-2.5 kW/m2K for these experiments. The results shown here suggest that in order to exploit the heat-transfer mediated freezing delay, thicker superhydrophobic coatings must be deposited on the surface, where the coating resistance is comparable to the bulk water/ice conduction resistance.

Porous and strong bioactive glass (13–93) scaffolds prepared by unidirectional freezing of camphene-based suspensions

PubMed Central

Liu, Xin; Rahaman, Mohamed N.; Fu, Qiang; Tomsia, Antoni P.

2011-01-01

Scaffolds of 13–93 bioactive glass (6Na2O, 12K2O, 5MgO, 20CaO, 4P2O5, 53SiO2; wt %) with an oriented pore architecture were formed by unidirectional freezing of camphene-based suspensions, followed by thermal annealing of the frozen constructs to grow the camphene crystals. After sublimation of the camphene, the constructs were sintered (1 h at 700 °C) to produce a dense glass phase with oriented macropores. The objective of this work was to study how constant freezing rates (1–7 °C/min) during the freezing step influenced the pore orientation and mechanical response of the scaffolds. When compared to scaffolds prepared by freezing the suspensions on a substrate kept at a constant temperature of 3 °C (time-dependent freezing rate), higher freezing rates resulted in better pore orientation, a more homogeneous microstructure, and a marked improvement in the mechanical response of the scaffolds in compression. Scaffolds fabricated using a constant freezing rate of 7 °C/min (porosity = 50 ± 4%; average pore diameter = 100 μm), had a compressive strength of 47 ± 5 MPa and an elastic modulus of 11 ± 3 GPa (in the orientation direction). In comparison, scaffolds prepared by freezing on the constant-temperature substrate had strength and modulus values of 35 ± 11 MPa and 8 ± 3 GPa, respectively. These oriented bioactive glass scaffolds prepared by the constant freezing rate route could potentially be used for the repair of defects in load-bearing bones, such as segmental defects in the long bones. PMID:21855661

Mutagenic effect of freezing on nuclear DNA of Saccharomyces cerevisiae.

PubMed

Todorova, T; Pesheva, M; Stamenova, R; Dimitrov, M; Venkov, P

2012-05-01

Although fragmentation of DNA has been observed in cells undergoing freezing procedures, a mutagenic effect of sub-zero temperature treatment has not been proved by induction and isolation of mutants in nuclear DNA (nDNA). In this communication we supply evidence for mutagenicity of freezing on nDNA of Saccharomyces cerevisiae cells. In the absence of cryoprotectors, cooling for 2 h at +4°C and freezing for 1 h at -10°C and 16 h at -20°C, with a cooling rate of 3°C/min, resulted in induction of frame-shift and reverse mutations in microsatellite and coding regions of nDNA. The sub-zero temperature exposure also has a strong recombinogenic effect, evidenced by induction of gene-conversion and crossing-over events. Freezing induces mutations and enhances recombination with a frequency equal to or higher than that of methylmethanesulphonate at comparable survival rates. The signals for the appearance of nDNA lesions induced by freezing are detected and transduced by the DNA damage pathway. Extracellular cryoprotectors did not prevent the mutagenic effect of freezing, while accumulation of trehalose inside cells reduced nDNA cryodamage. Freezing of cells is accompanied by generation of high ROS levels, and the oxidative stress raised during the freeze-thaw process is the most likely reason for the DNA damaging effect. Experiments with mitochondrial rho⁻ mutants or scavengers of ROS indicated that mutagenic and recombinogenic effects of sub-zero temperatures can be decreased but not eliminated by reduction of ROS level. The complete protection against cryodamage in nDNA required simultaneous usage of intracellular cryoprotector and ROS scavenger during the freeze-thaw process. Copyright © 2012 John Wiley & Sons, Ltd.

Effects of a controlled freeze-thaw event on dissolved and colloidal soil organic matter.

PubMed

Kim, Eun-Ah; Lee, Ha Kyung; Choi, Jung Hyun

2017-01-01

This study investigated the effects of the freezing and thawing that accompany the warming process on the composition of the soil organic matter in the dissolved and colloidal fractions. Temperate soil samples were incubated in a refrigerator at 2 °C for 4 weeks and compared with those frozen at -20 °C in the second week followed by thawing at 2 °C to study a freeze-thaw effect with minimal effect from the thawing temperature. The freeze-thaw group was compared with those incubated at 25 °C in the last week to investigate a warming effect after thawing. Thawing at 2 °C after freezing at -20 °C increased the dissolved organic carbon (DOC), but decreased colloidal Ca. The subsequent warming condition greatly increased both DOC and colloidal Ca. The colloidal organic carbon (COC) and dissolved Ca showed rather subtle changes in response to the freeze-thaw and warming treatments compared to the changes in DOC and colloidal Ca. The fluorescence excitation-emission matrix (EEM) and Fourier transformation-infrared spectrometry (FT-IR) results showed that the freeze-thaw and warming treatments gave the opposite effects on the compositions of dissolved humic-like substances, polysaccharides or silicates, and aliphatic alcohols. A principal component analysis (PCA) with the DOC, fluorescence EEM, and FT-IR spectra produced two principal components that successfully distinguished the effects of the freeze-thaw and warming treatments. Due to the contrasting effects of the freeze-thaw and warming treatments, the overall effects of freeze-thaw events in nature on the dissolved and colloidal soil organic matter could vary depending on the thawing temperature.

Monitoring Freeze Injury and Evaluating Losingto Sugar-Cane Using RS and GPS

NASA Astrophysics Data System (ADS)

Tan, Zongkun; Ding, Meihua; Wang, Longhe; Yang, Xin; Ou, Zhaorong

From Jan 12th to Feb 12th 2008, the most severity cold chilling and freeze injury weather took place during the last 50 years in the southern of China.Sugar-cane was suffered injury severity. However, the losing of sugar-cane which it was aroused by thisweather disaster had not been exactitude evaluated till on Apr 1st, 2008. It was not only affected the sugar-cane ordinary harvesting and crushing, but also affected reserving sugar-cane seed for planting. Freeze injury is common disaster for sugar-cane in southern of China and monitoring freeze injury using RS and GIS are of great economic significance but little research work about it has been done in China Freeze injuring is not only related to crop growth stage and the cold air intension from northern to southern and weather types, but also consanguineous related to land form and physiognomy and geographical latitude and height above sea level etc and crop planting spatial distribution. The case study of Guangxi province which is the biggest region of sugar-cane planting in China in this paper, the values of sugar-cane NDVI among the freeze injury occur former and after in early 2008 and without freeze injury occur in the same term 2007 were analyzed and compared based on the sugar-cane planting spatial distribution information which were carried out by using multi-phase EOS/MODIS data. The result showed that it was not only commendably reflected the spatial distribution of freeze injury but also reflected the sugarcane suffered from degree using the values of sugar-cane NDVIof freeze injury occur former and after. The field sample investigation data of using GPS was integrated with the NDVI, the evaluation of region sugar-cane suffer from freeze injury losing could quickly and exactly realize.

Comparative transcriptome profiling of freezing stress responses in loquat (Eriobotrya japonica) fruitlets.

PubMed

Xu, Hong-Xia; Li, Xiao-Ying; Chen, Jun-Wei

2017-09-01

Loquat (Eriobotrya japonica Lindl.) is an important subtropical, commercial fruit in China. It blossoms during autumn and winter in most areas of China and its fruitlets usually suffer from freezing stress. However, studies about the mechanisms underlying freezing stress in loquat are very limited. The gene expression profiles of loquat fruitlets subjected to freezing (G2 library) versus non-treated ones (G1 library) were investigated using Illumina sequencing technology to elucidate the molecular mechanisms and identify the genes that play vital roles in the freezing stress response. The results showed that approximately 157.63 million reads in total were obtained from freeze-treated and non-treated loquat fruitlets. These reads were assembled into 87,379 unigenes with an average length of 710 bp and an N50 of 1,200 bp. After comparing the profiles obtained from the G1 and G2 libraries, 2,892 differentially expressed genes were identified, of which 1,883 were up-regulated and 1,009 were down-regulated in the treated samples compared to non-treated ones. These unigenes showed significant differences in expression for carbohydrate transport and metabolism, amino acid metabolism, energy metabolism, and lipid metabolism, which are involved in defense against freezing stress. Glycolysis/gluconeogenesis was one of the most significantly regulated pathways. Freezing also significantly damaged the membrane system of loquat fruitlets, and several defense mechanisms were induced. Some selected genes related to low temperature resistance were validated by quantitative real-time PCR (qRT-PCR). The results revealed many genes and pathways that are part of freezing resistance processes and expand our understanding of the complex molecular events involved in freezing stress.

Infective Juveniles of the Entomopathogenic Nematode, Steinernema feltiae Produce Cryoprotectants in Response to Freezing and Cold Acclimation

PubMed Central

Ali, Farman; Wharton, David A.

2015-01-01

Steinernema feltiae is a moderately freeze-tolerant entomopathogenic nematode which survives intracellular freezing. We have detected by gas chromatography that infective juveniles of S. feltiae produce cryoprotectants in response to cold acclimation and to freezing. Since the survival of this nematode varies with temperature, we analyzed their cryoprotectant profiles under different acclimation and freezing regimes. The principal cryoprotectants detected were trehalose and glycerol with glucose being the minor component. The amount of cryoprotectants varied with the temperature and duration of exposure. Trehalose was accumulated in higher concentrations when nematodes were acclimated at 5°C for two weeks whereas glycerol level decreased from that of the non-acclimated controls. Nematodes were seeded with a small ice crystal and held at -1°C, a regime that does not produce freezing of the nematodes but their bodies lose water to the surrounding ice (cryoprotective dehydration). This increased the levels of both trehalose and glycerol, with glycerol reaching a higher concentration than trehalose. Nematodes frozen at -3°C, a regime that produces freezing of the nematodes and results in intracellular ice formation, had elevated glycerol levels while trehalose levels did not change. Steinernema feltiae thus has two strategies of cryoprotectant accumulation: one is an acclimation response to low temperature when the body fluids are in a cooled or supercooled state and the infective juveniles produce trehalose before freezing. During this process a portion of the glycerol is converted to trehalose. The second strategy is a rapid response to freezing which induces the production of glycerol but trehalose levels do not change. These low molecular weight compounds are surmised to act as cryoprotectants for this species and to play an important role in its freezing tolerance. PMID:26509788

Enzymatic Regulation of Glycogenolysis in a Subarctic Population of the Wood Frog: Implications for Extreme Freeze Tolerance

PubMed Central

do Amaral, M. Clara F.; Lee, Richard E.; Costanzo, Jon P.

2013-01-01

The wood frog, Rana sylvatica, from Interior Alaska survives freezing at –16°C, a temperature 10–13°C below that tolerated by its southern conspecifics. We investigated the hepatic freezing response in this northern phenotype to determine if its profound freeze tolerance is associated with an enhanced glucosic cryoprotectant system. Alaskan frogs had a larger liver glycogen reserve that was mobilized faster during early freezing as compared to conspecifics from a cool-temperate region (southern Ohio, USA). In Alaskan frogs the rapid glucose production in the first hours of freezing was associated with a 7-fold increase in glycogen phosphorylase activity above unfrozen frog levels, and the activity of this enzyme was higher than that of frozen Ohioan frogs. Freezing of Ohioan frogs induced a more modest (4-fold) increase in glycogen phosphorylase activity above unfrozen frog values. Relative to the Ohioan frogs, Alaskan frogs maintained a higher total protein kinase A activity throughout an experimental freezing/thawing time course, and this may have potentiated glycogenolysis during early freezing. We found populational variation in the activity and protein level of protein kinase A which suggested that the Alaskan population had a more efficient form of this enzyme. Alaskan frogs modulated their glycogenolytic response by decreasing the activity of glycogen phosphorylase after cryoprotectant mobilization was well under way, thereby conserving their hepatic glycogen reserve. Ohioan frogs, however, sustained high glycogen phosphorylase activity until early thawing and consumed nearly all their liver glycogen. These unique hepatic responses of Alaskan R. sylvatica likely contribute to this phenotype’s exceptional freeze tolerance, which is necessary for their survival in a subarctic climate. PMID:24236105

Preservation of flavor in freeze dried green beans

NASA Technical Reports Server (NTRS)

Huber, C. S.; Heidelbaugh, N. D.; Davis, D.

1973-01-01

Before freeze drying, green beans are heated to point at which their cell structure is altered. Beans freeze dried with altered cell structure have improved rehydration properties and retain color, flavor, and texture.

9 CFR 590.536 - Freezing operations.

Code of Federal Regulations, 2012 CFR

2012-01-01

..., and Facility Requirements § 590.536 Freezing operations. (a) Freezing rooms shall be kept clean and... outside of liquid egg containers shall be clean and free from evidence of liquid egg. (e) Frozen egg...

9 CFR 590.536 - Freezing operations.

Code of Federal Regulations, 2014 CFR

2014-01-01

..., and Facility Requirements § 590.536 Freezing operations. (a) Freezing rooms shall be kept clean and... outside of liquid egg containers shall be clean and free from evidence of liquid egg. (e) Frozen egg...

9 CFR 590.536 - Freezing operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

..., and Facility Requirements § 590.536 Freezing operations. (a) Freezing rooms shall be kept clean and... outside of liquid egg containers shall be clean and free from evidence of liquid egg. (e) Frozen egg...

9 CFR 590.536 - Freezing operations.

Code of Federal Regulations, 2013 CFR

2013-01-01

..., and Facility Requirements § 590.536 Freezing operations. (a) Freezing rooms shall be kept clean and... outside of liquid egg containers shall be clean and free from evidence of liquid egg. (e) Frozen egg...

Freeze-In dark matter with displaced signatures at colliders

DOE Office of Scientific and Technical Information (OSTI.GOV)

Co, Raymond T.; D’Eramo, Francesco; Hall, Lawrence J.

2015-12-11

Dark matter, X, may be generated by new physics at the TeV scale during an early matter-dominated (MD) era that ends at temperature T{sub R}≪ TeV. Compared to the conventional radiation-dominated (RD) results, yields from both Freeze-Out and Freeze-In processes are greatly suppressed by dilution from entropy production, making Freeze-Out less plausible while allowing successful Freeze-In with a much larger coupling strength. Freeze-In is typically dominated by the decay of a particle B of the thermal bath, B→X. For a large fraction of the relevant cosmological parameter space, the decay rate required to produce the observed dark matter abundance leadsmore » to displaced signals at LHC and future colliders, for any m{sub X} in the range keV« less

Effect of ultrasonic waves on the freezing rates of potatoes in degassed coolant and untreated coolant.

PubMed

Yu, D Y; Liu, B L

2014-01-01

Ultrasonic waves are shown to enhance the rate of freezing. To elucidate the mechanism of immersion freezing of potatos with ultrasonic waves. Ultrasound is applied to potato samples immersed in degassed coolant and untreated coolant. Sonic waves were intermittently applied at temperature below -1 degree C. The freezing rates were measured under different experimental conditions. The use of ultrasonic waves increased the freezing rates of potatoes immersed in both degassed coolant and untreated coolant. However, the freezing rate in the degassed coolant was less than that in the untreated coolant. Heat transfer on the interface between the potato sample and sonicated degassed coolant appears to be less than that within the sample in the absence of cavitation. The interface heat transfer between the potato sample and untreated coolant is likely improved due to ultrasonic cavitation.

Photomicrographic Investigation of Spontaneous Freezing Temperatures of Supercooled Water Droplets

NASA Technical Reports Server (NTRS)

Dorsch, R. G.; Hacker, P. T.

1950-01-01

A photomicrographic technique for investigating eupercooled. water droplets has been devised and. used. to determine the spontaneous freezing temperatures of eupercooled. water droplets of the size ordinarily found. in the atmosphere. The freezing temperatures of 4527 droplets ranging from 8.75 to 1000 microns in diameter supported on a platinum surface and 571 droplets supported on copper were obtained. The average spontaneous freezing temperature decreased with decrease in the size of the droplets. The effect of size on the spontaneous freezing temperature was particularly marked below 60 microns. Frequency-distribution curves of the spontaneous freezing temperatures observed for droplets of a given size were obtained. Although no droplet froze at a temperature above 20 0 F, all droplets melted at 32 F. Results obtained with a copper support did not differ essentially from those obtained with a platinum surface.

Design of freeze-drying processes for pharmaceuticals: practical advice.

PubMed

Tang, Xiaolin; Pikal, Michael J

2004-02-01

Design of freeze-drying processes is often approached with a "trial and error" experimental plan or, worse yet, the protocol used in the first laboratory run is adopted without further attempts at optimization. Consequently, commercial freeze-drying processes are often neither robust nor efficient. It is our thesis that design of an "optimized" freeze-drying process is not particularly difficult for most products, as long as some simple rules based on well-accepted scientific principles are followed. It is the purpose of this review to discuss the scientific foundations of the freeze-drying process design and then to consolidate these principles into a set of guidelines for rational process design and optimization. General advice is given concerning common stability issues with proteins, but unusual and difficult stability issues are beyond the scope of this review. Control of ice nucleation and crystallization during the freezing step is discussed, and the impact of freezing on the rest of the process and final product quality is reviewed. Representative freezing protocols are presented. The significance of the collapse temperature and the thermal transition, denoted Tg', are discussed, and procedures for the selection of the "target product temperature" for primary drying are presented. Furthermore, guidelines are given for selection of the optimal shelf temperature and chamber pressure settings required to achieve the target product temperature without thermal and/or mass transfer overload of the freeze dryer. Finally, guidelines and "rules" for optimization of secondary drying and representative secondary drying protocols are presented.

Optimum Parameters for Freeze-Drying Decellularized Arterial Scaffolds

PubMed Central

Sheridan, William S.; Duffy, Garry P.

2013-01-01

Decellularized arterial scaffolds have achieved success in advancing toward clinical use as vascular grafts. However, concerns remain regarding long-term preservation and sterilization of these scaffolds. Freeze drying offers a means of overcoming these concerns. In this study, we investigated the effects of various freeze-drying protocols on decellularized porcine carotid arteries and consequently, determined the optimum parameters to fabricate a stable, preserved scaffold with unaltered mechanical properties. Freeze drying by constant slow cooling to two final temperatures ((Tf), −10°C and −40°C) versus instant freezing was investigated by histological examination and mechanical testing. Slow cooling to Tf= −10°C produced a stiffer and less distensible response than the non freeze-dried scaffolds and resulted in disruption to the collagen fibers. The mechanical response of Tf= −40°C scaffolds demonstrated disruption to the elastin network, which was confirmed with histology. Snap freezing scaffolds in liquid nitrogen and freeze drying to Tf= −40°C with a precooled shelf at −60°C produced scaffolds with unaltered mechanical properties and a histology resembling non-freeze-dried scaffolds. The results of this study demonstrate the importance of optimizing the nucleation and ice crystal growth/size to ensure homogenous drying, preventing extracellular matrix disruption and subsequent inferior mechanical properties. This new manufacturing protocol creates the means for the preservation and sterilization of decellularized arterial scaffolds while simultaneously maintaining the mechanical properties of the tissue. PMID:23614758

Validation of the shake test for detecting freeze damage to adsorbed vaccines.

PubMed

Kartoglu, Umit; Ozgüler, Nejat Kenan; Wolfson, Lara J; Kurzatkowski, Wiesław

2010-08-01

To determine the validity of the shake test for detecting freeze damage in aluminium-based, adsorbed, freeze-sensitive vaccines. A double-blind crossover design was used to compare the performance of the shake test conducted by trained health-care workers (HCWs) with that of phase contrast microscopy as a "gold standard". A total of 475 vials of 8 different types of World Health Organization prequalified freeze-sensitive vaccines from 10 different manufacturers were used. Vaccines were kept at 5 degrees C. Selected numbers of vials from each type were then exposed to -25 degrees C and -2 degrees C for 24-hour periods. There was complete concordance between HCWs and phase-contrast microscopy in identifying freeze-damaged vials and non-frozen samples. Non-frozen samples showed a fine-grain structure under phase contrast microscopy, but freeze-damaged samples showed large conglomerates of massed precipitates with amorphous, crystalline, solid and needle-like structures. Particles in the non-frozen samples measured from 1 microm (vaccines against diphtheria-tetanus-pertussis; Haemophilus influenzae type b; hepatitis B; diphtheria-tetanus-pertussis-hepatitis B) to 20 microm (diphtheria and tetanus vaccines, alone or in combination). By contrast, aggregates in the freeze-damaged samples measured up to 700 microm (diphtheria-tetanus-pertussis) and 350 microm on average. The shake test had 100% sensitivity, 100% specificity and 100% positive predictive value in this study, which confirms its validity for detecting freeze damage to aluminium-based freeze-sensitive vaccines.

Model for heat and mass transfer in freeze-drying of pellets.

PubMed

Trelea, Ioan Cristian; Passot, Stéphanie; Marin, Michèle; Fonseca, Fernanda

2009-07-01

Lyophilizing frozen pellets, and especially spray freeze-drying, have been receiving growing interest. To design efficient and safe freeze-drying cycles, local temperature and moisture content in the product bed have to be known, but both are difficult to measure in the industry. Mathematical modeling of heat and mass transfer helps to determine local freeze-drying conditions and predict effects of operation policy, and equipment and recipe changes on drying time and product quality. Representative pellets situated at different positions in the product slab were considered. One-dimensional transfer in the slab and radial transfer in the pellets were assumed. Coupled heat and vapor transfer equations between the temperature-controlled shelf, the product bulk, the sublimation front inside the pellets, and the chamber were established and solved numerically. The model was validated based on bulk temperature measurement performed at two different locations in the product slab and on partial vapor pressure measurement in the freeze-drying chamber. Fair agreement between measured and calculated values was found. In contrast, a previously developed model for compact product layer was found inadequate in describing freeze-drying of pellets. The developed model represents a good starting basis for studying freeze-drying of pellets. It has to be further improved and validated for a variety of product types and freeze-drying conditions (shelf temperature, total chamber pressure, pellet size, slab thickness, etc.). It could be used to develop freeze-drying cycles based on product quality criteria such as local moisture content and glass transition temperature.

Freezing Nitrogen Ethanol Composite May be a Viable Approach for Cryotherapy of Human Giant Cell Tumor of Bone.

PubMed

Wu, Po-Kuei; Chen, Cheng-Fong; Wang, Jir-You; Chen, Paul Chih-Hsueh; Chang, Ming-Chau; Hung, Shih-Chieh; Chen, Wei-Ming

2017-06-01

Liquid nitrogen has been used as adjuvant cryotherapy for treating giant cell tumor (GCT) of bone. However, the liquid phase and ultrafreezing (-196° C) properties increase the risk of damage to the adjacent tissues and may lead to perioperative complications. A novel semisolid cryogen, freezing nitrogen ethanol composite, might mitigate these shortcomings because of less-extreme freezing. We therefore wished to evaluate freezing nitrogen ethanol composite as a coolant to determine its properties in tumor cryoablation. (1) Is freezing nitrogen ethanol composite-mediated freezing effective for tumor cryoablation in an ex vivo model, and if yes, is apoptosis involved in the tumor-killing mechanism? (2) Does freezing nitrogen ethanol composite treatment block neovascularization and neoplastic progression of the grafted GCTs and is it comparable to that of liquid nitrogen in an in vivo chicken model? (3) Can use of freezing nitrogen ethanol composite as an adjuvant to curettage result in successful short-term treatment, defined as absence of GCT recurrence at a minimum of 1 year in a small proof-of-concept clinical series? The cryogenic effect on bone tissue mediated by freezing nitrogen ethanol composite and liquid nitrogen was verified by thermal measurement in a time-course manner. Cryoablation on human GCT tissue was examined ex vivo for effect on morphologic features (cell shrinkage) and DNA fragmentation (apoptosis). The presumed mechanism was investigated by molecular analysis of apoptosis regulatory proteins including caspases 3, 8, and 9 and Bax/Bcl-2. Chicken chorioallantoic membrane was used as an in vivo model to evaluate the effects of freezing nitrogen ethanol composite and liquid nitrogen treatment on GCT-derived neovascularization and tumor neoplasm. A small group of patients with GCT of bone was treated by curettage and adjuvant freezing nitrogen ethanol composite cryotherapy in a proof-of-concept study. Tumor recurrence and perioperative complications were evaluated at a minimum of 19 months followup (mean, 24 months; range, 19-30 months). Freshly prepared freezing nitrogen ethanol composite froze to -136° C and achieved -122° C isotherm across a piece of 10 ± 0.50-mm-thick bone with a freezing rate of -34° C per minute, a temperature expected to meet clinical tumor-killing requirements. Human GCT tissues revealed histologic changes including shrinkage in morphologic features of multinucleated giant cells in the liquid nitrogen (202 ± 45 μm; p = 0.006) and freezing nitrogen ethanol composite groups (169 ± 27.4 μm; p < 0.001), and a decreased nucleated area of neoplastic stromal cells for the 30-second treatment. Enhanced counts of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells verified the involvement of DNA fragmentation in cryoablated GCT tissues. Western blotting analysis on the expression of apoptosis regulatory proteins showed enhancement of proteocleavage-activated caspases 3, 8, and 9 and higher ratios of Bax/Bcl2 in the liquid nitrogen- and freezing nitrogen ethanol composite-treated samples. Numbers of blood vessels and human origin tumor cells also were decreased by freezing nitrogen ethanol composite and liquid nitrogen treatment in the GCT-grafted chicken chorioallantoic membrane model. Seven patients with GCT treated by curettage and adjuvant cryotherapy by use of freezing nitrogen ethanol composite preparation had no intra- or postoperative complications related to the freezing, and no recurrences during the study surveillance period. These preliminary in vitro and clinical findings suggest that freezing nitrogen ethanol composite may be an effective cryogen showing ex vivo and in vivo tumor cryoablation comparable to liquid nitrogen. The semisolid phase and proper thermal conduction might avoid some of the disadvantages of liquid nitrogen in cryotherapy, but a larger clinical study is needed to confirm these findings. Level IV, therapeutic study.

Freeze-thaw-induced embolism in Pinus contorta: centrifuge experiments validate the 'thaw-expansion hypothesis' but conflict with ultrasonic emission data.

PubMed

Mayr, Stefan; Sperry, John S

2010-03-01

*The 'thaw-expansion hypothesis' postulates that xylem embolism is caused by the formation of gas bubbles on freezing and their expansion on thawing. We evaluated the hypothesis using centrifuge experiments and ultrasonic emission monitoring in Pinus contorta. *Stem samples were exposed to freeze-thaw cycles at varying xylem pressure (P) in a centrifuge before the percentage loss of hydraulic conductivity (PLC) was measured. Ultrasonic acoustic emissions were registered on samples exposed to freeze-thaw cycles in a temperature chamber. *Freeze-thaw exposure of samples spun at -3 MPa induced a PLC of 32% (one frost cycle) and 50% (two cycles). An increase in P to -0.5 MPa during freezing had no PLC effect, whereas increased P during thaw lowered PLC to 7%. Ultrasonic acoustic emissions were observed during freezing and thawing at -3 MPa, but not in air-dried or water-saturated samples. A decrease in minimum temperature caused additional ultrasonic acoustic emissions, but had no effect on PLC. *The centrifuge experiments indicate that the 'thaw-expansion hypothesis' correctly describes the embolization process. Possible explanations for the increase in PLC on repeated frost cycles and for the ultrasonic acoustic emissions observed during freezing and with decreasing ice temperature are discussed.

Apoptosis-like death was involved in freeze-drying-preserved fungus Mucor rouxii and can be inhibited by L-proline.

PubMed

Wang, Xiaoyun; Wang, Youzhi

2016-02-01

Freeze-drying is one of the most effective methods to preserve fungi for an extended period. However, it is associated with a loss of viability and shortened storage time in some fungi. This study evaluated the stresses that led to the death of freeze-dried Mucor rouxii by using cell apoptotic methods. The results showed there were apoptosis-inducing stresses, such as the generation of obvious intracellular reactive oxygen species (ROS) and metacaspase activation. Moreover, nuclear condensation and a delayed cell death peak were determined after rehydration and 24 h incubation in freeze-dried M. rouxii via a propidium iodide (PI) assay, which is similar to the phenomenon of cryopreservation-induced delayed-onset cell death (CIDOCD). Then, several protective agents were tested to decrease the apoptosis-inducing stresses and to improve the viability. Finally, it was found that 1.6 mM L-proline can effectively decrease the nuclear condensation rate and increase the survival rate in freeze-dried M. rouxii. (1) apoptosis-inducing factors occur in freeze-dried M. rouxii. (2) ROS and activated metacaspases lead to death in freeze-dried M. rouxii. (3)L-proline increases the survival rate of freeze-dried M. rouxii. Copyright © 2015 Elsevier Inc. All rights reserved.

Effects of annealing on the physical properties of therapeutic proteins during freeze drying process.

PubMed

Lim, Jun Yeul; Lim, Dae Gon; Kim, Ki Hyun; Park, Sang-Koo; Jeong, Seong Hoon

2018-02-01

Effects of annealing steps during the freeze drying process on etanercept, model protein, were evaluated using various analytical methods. The annealing was introduced in three different ways depending on time and temperature. Residual water contents of dried cakes varied from 2.91% to 6.39% and decreased when the annealing step was adopted, suggesting that they are directly affected by the freeze drying methods Moreover, the samples were more homogenous when annealing was adopted. Transition temperatures of the excipients (sucrose, mannitol, and glycine) were dependent on the freeze drying steps. Size exclusion chromatography showed that monomer contents were high when annealing was adopted and also they decreased less after thermal storage at 60°C. Dynamic light scattering results exhibited that annealing can be helpful in inhibiting aggregation and that thermal storage of freeze-dried samples preferably induced fragmentation over aggregation. Shift of circular dichroism spectrum and of the contents of etanercept secondary structure was observed with different freeze drying steps and thermal storage conditions. All analytical results suggest that the physicochemical properties of etanercept formulation can differ in response to different freeze drying steps and that annealing is beneficial for maintaining stability of protein and reducing the time of freeze drying process. Copyright © 2017 Elsevier B.V. All rights reserved.

Hatchling painted turtles (Chrysemys picta) survive only brief freezing of their bodily fluids.

PubMed

Attaway, M B; Packard, G C; Packard, M J

1998-07-01

Neonatal painted turtles (Chrysemys picta) spend their first winter inside the shallow, subterranean nest cavity where they completed embryogenesis. Consequently, hatchlings at high latitudes may be exposed to ice and cold during the winter. This study was undertaken to determine how long hatchlings withstand freezing at temperatures slightly below 0 degree C because tolerance for freezing has been proposed to be the key to survival by overwintering animals. A thermocouple was glued to the carapace of each hatchling. The animal was dipped in water to provide a site of nucleation of ice and was then placed into a glass jar that was partially immersed in a circulating bath at -2 degrees C. Carapace temperature was monitored throughout the procedure. When a freezing exotherm was detected, timing of the freezing event began. Animals were maintained in a frozen state for 12-48 h prior to being warmed to room temperature. Of the 39 hatchlings, 22 did not survive, and mortality increased as the duration of freezing increased. Logistic regression indicates that no turtle would have survived in a frozen state for more than 54 h. These results indicate that hatchlings can survive only brief exposure to freezing of the body fluids. Thus, hatchlings cannot tolerate freezing during prolonged periods of cold.

Metabolic Changes in Avena sativa Crowns Recovering from Freezing

PubMed Central

Henson, Cynthia A.; Duke, Stanley H.; Livingston, David P.

2014-01-01

Extensive research has been conducted on cold acclimation and freezing tolerance of fall-sown cereal plants due to their economic importance; however, little has been reported on the biochemical changes occurring over time after the freezing conditions are replaced by conditions favorable for recovery and growth such as would occur during spring. In this study, GC-MS was used to detect metabolic changes in the overwintering crown tissue of oat (Avena sativa L.) during a fourteen day time-course after freezing. Metabolomic analysis revealed increases in most amino acids, particularly proline, 5-oxoproline and arginine, which increased greatly in crowns that were frozen compared to controls and correlated very significantly with days after freezing. In contrast, sugar and sugar related metabolites were little changed by freezing, except sucrose and fructose which decreased dramatically. In frozen tissue all TCA cycle metabolites, especially citrate and malate, decreased in relation to unfrozen tissue. Alterations in some amino acid pools after freezing were similar to those observed in cold acclimation whereas most changes in sugar pools after freezing were not. These similarities and differences suggest that there are common as well as unique genetic mechanisms between these two environmental conditions that are crucial to the winter survival of plants. PMID:24675792

Simulation of the process kinetics and analysis of physicochemical properties in the freeze drying of kale

NASA Astrophysics Data System (ADS)

Dziki, Dariusz; Polak, Renata; Rudy, Stanisław; Krzykowski, Andrzej; Gawlik-Dziki, Urszula; Różyło, Renata; Miś, Antoni; Combrzyński, Maciej

2018-01-01

Investigations were performed to study the freeze-drying process of kale (Brassica oleracea L. var acephala). The process of freeze-drying was performed at temperatures of 20, 40, and 60°C for whole pieces of leaves and for pulped leaves. The kinetics of the freeze-drying of both kale leaves and kale pulp were best described by the Page model. The increasing freeze-drying temperature from 20 to 60°C induced an approximately two-fold decrease in the drying time. Freeze-drying significantly increased the value of the lightness, delta Chroma, and browning index of kale, and had little influence on the hue angle. The highest increase in the lightness and delta Chroma was observed for whole leaves freeze-dried at 20°C. An increase in the drying temperature brought about a slight decrease in the lightness, delta Chroma and the total colour difference. Pulping decreased the lightness and hue angle, and increased browning index. Freeze-drying engendered a slight decrease in the total phenolics content and antioxidant activity, in comparison to fresh leaves. The temperature of the process and pulping had little influence on the total phenolics content and antioxidant activity of dried kale, but significantly decreased the contents of chlorophyll a and chlorophyll b.

Characterization of Propylene Glycol-Mitigated Freeze/Thaw Agglomeration of a Frozen Liquid nOMV Vaccine Formulation by Static Light Scattering and Micro-Flow Imaging.

PubMed

Mensch, Christopher D; Davis, Harrison B; Blue, Jeffrey T

2015-01-01

The purpose of this work was to investigate the susceptibility of an aluminum adjuvant and an aluminum-adjuvanted native outer membrane vesicle (nOMV) vaccine formulation to freeze/thaw-induced agglomeration using static light scattering and micro-flow Imaging analysis; and to evaluate the use of propylene glycol as a vaccine formulation excipient by which freeze/thaw-induced agglomeration of a nOMV vaccine formulation could be mitigated. Our results indicate that including 7% v/v propylene glycol in an nOMV containing aluminum adjuvanted vaccine formulation, mitigates freeze/thaw-induced agglomeration. We evaluated the effect of freeze-thawing on an aluminum adjuvant and an aluminum adjuvanted native outer membrane vesicle (nOMV) vaccine formulation. Specifically, we characterized the freeze/thaw-induced agglomeration through the use of static light scattering, micro-flow imaging, and cryo-electron microscopy analysis. Further, we evaluated the use of 0-9% v/v propylene glycol as an excipient which could be included in the formulation for the purpose of mitigating the agglomeration induced by freeze/thaw. The results indicate that using 7% v/v propylene glycol as a formulation excipient is effective at mitigating agglomeration of the nOMV vaccine formulation, otherwise induced by freeze-thawing. © PDA, Inc. 2015.

Effect of Novel Quick Freezing Techniques Combined with Different Thawing Processes on Beef Quality

PubMed Central

Yoo, Seon-Mi; Han, Gui-Jung

2014-01-01

This study investigated the effect of various freezing and thawing techniques on the quality of beef. Meat samples were frozen using natural convection freezing (NF), individual quick freezing (IQF), or cryogenic freezing (CF) techniques, followed by natural convection thawing (NCT) or running water thawing (RT). The meat was frozen until the core temperature reached -12℃ and then stored at -24℃, followed by thawing until the temperature reached 5℃. Quality parameters, such as the pH, water binding properties, CIE color, shear force, and microstructure of the beef were elucidated. Although the freezing and thawing combinations did not cause remarkable changes in the quality parameters, rapid freezing, in the order of CF, IQF, and NF, was found to minimize the quality deterioration. In the case of thawing methods, NCT was better than RT and the meat quality was influence on the thawing temperature rather than the thawing rate. Although the microstructure of the frozen beef exhibited an excessive loss of integrity after the freezing and thawing, it did not cause any remarkable change in the beef quality. Taken together, these results demonstrate that CF and NCT form the best combination for beef processing; however, IQF and NCT may have practical applications in the frozen food industry. PMID:26761674

Monitoring of Freezing Dynamics in Trees: A Simple Phase Shift Causes Complexity1[OPEN

PubMed Central

Charra-Vaskou, Katline

2017-01-01

During winter, trees have to cope with harsh conditions, including extreme freeze-thaw stress. This study focused on ice nucleation and propagation, related water shifts and xylem cavitation, as well as cell damage and was based on in situ monitoring of xylem (thermocouples) and surface temperatures (infrared imaging), ultrasonic emissions, and dendrometer analysis. Field experiments during late winter on Picea abies growing at the alpine timberline revealed three distinct freezing patterns: (1) from the top of the tree toward the base, (2) from thin branches toward the main stem’s top and base, and (3) from the base toward the top. Infrared imaging showed freezing within branches from their base toward distal parts. Such complex freezing causes dynamic and heterogenous patterns in water potential and probably in cavitation. This study highlights the interaction between environmental conditions upon freezing and thawing and demonstrates the enormous complexity of freezing processes in trees. Diameter shrinkage, which indicated water fluxes within the stem, and acoustic emission analysis, which indicated cavitation events near the ice front upon freezing, were both related to minimum temperature and, upon thawing, related to vapor pressure deficit and soil temperature. These complex patterns, emphasizing the common mechanisms between frost and drought stress, shed new light on winter tree physiology. PMID:28242655

Folic acid content in thermostabilized and freeze-dried space shuttle foods

NASA Technical Reports Server (NTRS)

Lane, H. W.; Nillen, J. L.; Kloeris, V. L.

1995-01-01

This study was designed to determine whether freeze-dried and thermostabilized foods on a space shuttle contain adequate folate and to investigate any effects of freeze-drying on folacin. Frozen vegetables were analyzed after three states of processing: thawed; cooked; and rehydrated. Thermostabilized items were analyzed as supplied with no further processing. Measurable folate decreased in some freeze-dried vegetables and increased in others. Folacin content of thermostabilized food items was comparable with published values. We concluded that although the folacin content of some freeze-dried foods was low, adequate folate is available from the shuttle menu to meet RDA guidelines.

Survival differences among freeze-dried genetically engineered and wild-type bacteria.

PubMed Central

Israeli, E; Shaffer, B T; Hoyt, J A; Lighthart, B; Ganio, L M

1993-01-01

Because the death mechanisms of freeze-dried and air-dried bacteria are thought to be similar, freeze-drying was used to investigate the survival differences between potentially airborne genetically engineered microorganisms and their wild types. To this end, engineered strains of Escherichia coli and Pseudomonas syringae were freeze-dried and exposed to air, visible light, or both. The death rates of all engineered strains were significantly higher than those of their parental strains. Light and air exposure were found to increase the death rates of all strains. Application of death rate models to freeze-dried engineered bacteria to be released into the environment is discussed. PMID:8434925

The effect of variety and maturity on the quality of freeze-dried carrots. The effect of microwave blanching on the nutritional and textural quality of freeze-dried spinach

NASA Technical Reports Server (NTRS)

1979-01-01

Using carrots, the quality of freeze-dried products was studied to determine the optimum varieties and maturation stages for quality attributes such as appearance, flavor, texture, and nutritive value. The quality of freeze-dried carrots is discussed in terms of Gardner color, alcohol insoluble solids, viscosity, and core/cortex ratio. Also, microwave blanching of freeze-dried spinach was studied to determine vitamin interrelationships, anatomical changes, and oxidative deteriorations in terms of preprocessing microwave treatments. Statistical methods were employed in the gathering of data and interpretation of results in both studies.

Topological Spin Glass in Diluted Spin Ice

NASA Astrophysics Data System (ADS)

Sen, Arnab; Moessner, R.

2015-06-01

It is a salient experimental fact that a large fraction of candidate spin liquid materials freeze as the temperature is lowered. The question naturally arises whether such freezing is intrinsic to the spin liquid ("disorder-free glassiness") or extrinsic, in the sense that a topological phase simply coexists with standard freezing of impurities. Here, we demonstrate a surprising third alternative, namely, that freezing and topological liquidity are inseparably linked. The topological phase reacts to the introduction of disorder by generating degrees of freedom of a new type (along with interactions between them), which in turn undergo a freezing transition while the topological phase supporting them remains intact.

Topological Spin Glass in Diluted Spin Ice.

PubMed

Sen, Arnab; Moessner, R

2015-06-19

It is a salient experimental fact that a large fraction of candidate spin liquid materials freeze as the temperature is lowered. The question naturally arises whether such freezing is intrinsic to the spin liquid ("disorder-free glassiness") or extrinsic, in the sense that a topological phase simply coexists with standard freezing of impurities. Here, we demonstrate a surprising third alternative, namely, that freezing and topological liquidity are inseparably linked. The topological phase reacts to the introduction of disorder by generating degrees of freedom of a new type (along with interactions between them), which in turn undergo a freezing transition while the topological phase supporting them remains intact.

A whole-plant screening test to identify genotypes with superior freezing tolerance.

PubMed

Bertrand, Annick; Castonguay, Yves; Bourassa, Josée

2014-01-01

Freezing tolerance is a determinant factor of persistence of perennials grown in northern climate. Selection for winterhardiness in field nurseries is difficult because of the unpredictability of occurrence of test winters allowing the identification of hardy genotypes. Here we describe a whole-plant assay entirely performed indoor in growth chambers and walk-in freezers to identify genotypes with superior tolerance to freezing within populations of open pollinated species. Three successive freezing stresses are applied to progressively eliminate 90 % of the population and to retain only the 10 % best performing genotypes. This approach can be used to generate recurrently selected populations more tolerant to freezing in different species.

Xylem Surfactants Introduce a New Element to the Cohesion-Tension Theory1[OPEN

PubMed Central

Espino, Susana; Nima, Neda; Do, Aissa Y.T.; Michaud, Joseph M.; Papahadjopoulos-Sternberg, Brigitte; Yang, Jinlong; Steppe, Kathy

2017-01-01

Vascular plants transport water under negative pressure without constantly creating gas bubbles that would disable their hydraulic systems. Attempts to replicate this feat in artificial systems almost invariably result in bubble formation, except under highly controlled conditions with pure water and only hydrophilic surfaces present. In theory, conditions in the xylem should favor bubble nucleation even more: there are millions of conduits with at least some hydrophobic surfaces, and xylem sap is saturated or sometimes supersaturated with atmospheric gas and may contain surface-active molecules that can lower surface tension. So how do plants transport water under negative pressure? Here, we show that angiosperm xylem contains abundant hydrophobic surfaces as well as insoluble lipid surfactants, including phospholipids, and proteins, a composition similar to pulmonary surfactants. Lipid surfactants were found in xylem sap and as nanoparticles under transmission electron microscopy in pores of intervessel pit membranes and deposited on vessel wall surfaces. Nanoparticles observed in xylem sap via nanoparticle-tracking analysis included surfactant-coated nanobubbles when examined by freeze-fracture electron microscopy. Based on their fracture behavior, this technique is able to distinguish between dense-core particles, liquid-filled, bilayer-coated vesicles/liposomes, and gas-filled bubbles. Xylem surfactants showed strong surface activity that reduces surface tension to low values when concentrated as they are in pit membrane pores. We hypothesize that xylem surfactants support water transport under negative pressure as explained by the cohesion-tension theory by coating hydrophobic surfaces and nanobubbles, thereby keeping the latter below the critical size at which bubbles would expand to form embolisms. PMID:27927981

Xylem Surfactants Introduce a New Element to the Cohesion-Tension Theory.

PubMed

Schenk, H Jochen; Espino, Susana; Romo, David M; Nima, Neda; Do, Aissa Y T; Michaud, Joseph M; Papahadjopoulos-Sternberg, Brigitte; Yang, Jinlong; Zuo, Yi Y; Steppe, Kathy; Jansen, Steven

2017-02-01

Vascular plants transport water under negative pressure without constantly creating gas bubbles that would disable their hydraulic systems. Attempts to replicate this feat in artificial systems almost invariably result in bubble formation, except under highly controlled conditions with pure water and only hydrophilic surfaces present. In theory, conditions in the xylem should favor bubble nucleation even more: there are millions of conduits with at least some hydrophobic surfaces, and xylem sap is saturated or sometimes supersaturated with atmospheric gas and may contain surface-active molecules that can lower surface tension. So how do plants transport water under negative pressure? Here, we show that angiosperm xylem contains abundant hydrophobic surfaces as well as insoluble lipid surfactants, including phospholipids, and proteins, a composition similar to pulmonary surfactants. Lipid surfactants were found in xylem sap and as nanoparticles under transmission electron microscopy in pores of intervessel pit membranes and deposited on vessel wall surfaces. Nanoparticles observed in xylem sap via nanoparticle-tracking analysis included surfactant-coated nanobubbles when examined by freeze-fracture electron microscopy. Based on their fracture behavior, this technique is able to distinguish between dense-core particles, liquid-filled, bilayer-coated vesicles/liposomes, and gas-filled bubbles. Xylem surfactants showed strong surface activity that reduces surface tension to low values when concentrated as they are in pit membrane pores. We hypothesize that xylem surfactants support water transport under negative pressure as explained by the cohesion-tension theory by coating hydrophobic surfaces and nanobubbles, thereby keeping the latter below the critical size at which bubbles would expand to form embolisms. © 2017 American Society of Plant Biologists. All Rights Reserved.

Breeding of Freeze-tolerant Yeast and the Mechanisms of Stress-tolerance

NASA Astrophysics Data System (ADS)

Hino, Akihiro

Frozen dough method have been adopted in the baking industry to reduce labor and to produce fresh breads in stores. New freeze-tolerant yeasts for frozen dough preparations were isolated from banana peel and identified. To obtain strains that have fermentative ability even after several months of frozen storage in fermented dough, we attempted to breed new freeze-tolerantstrain. The hybrid between S.cerevisiae, which is a isolated freeze-tolerant strain, and a strain isolated from bakers' yeast with sexual conjugation gave a good quality bread made from frozen dough method. Freeze-tolerant strains showed higher surviving and trehalose accumulating abilities than freeze-sensitive strains. The freeze tolerance of the yeasts was associated with the basal amount of intracellular trehalose after rapid degradation at the onset of the prefermentation period. The complicated metabolic pathway and the regulation system of trehalose in yeast cells are introduced. The trehalose synthesis may act as a metabolic buffer system which contribute to maintain the intracellular inorganic phosphate and as a feedback regulation system in the glycolysis. However, it is not known enough how the trehalose protects yeast cells from stress.

Freezing Directed Construction of Bio/Nano Interfaces: Reagentless Conjugation, Denser Spherical Nucleic Acids, and Better Nanoflares.

PubMed

Liu, Biwu; Liu, Juewen

2017-07-19

While nanoparticle solutions cannot freeze in general, they may remain stable in the presence of polymer stabilizers. We herein communicate that gold nanoparticles (AuNPs) are stable in the presence of thiolated DNA after a freeze-thaw cycle. The DNA is conjugated to AuNPs during freezing without additional reagents and the conjugation can be completed in a few minutes. More importantly, the DNA density is 20-30% higher than that prepared by the typical salt-aging method. By lowering temperature, DNA hybridization is also promoted, allowing the construction of better nanoflares with doubled probe density and signaling sensitivity. This freezing method works for AuNPs from 5 to 100 nm and all tested DNA sequences. The mechanism was studied by separating the effect of temperature, freezing and thawing, where the exclusion of salt and AuNPs by the growing ice crystals is deemed critical. In addition to developing a simple method, this study articulates unique physical processes during freezing with important fundamental surface science implications, and it could be extended to other systems.

Experimental realization of self-guided quantum coherence freezing

NASA Astrophysics Data System (ADS)

Yu, Shang; Wang, Yi-Tao; Ke, Zhi-Jin; Liu, Wei; Zhang, Wen-Hao; Chen, Geng; Tang, Jian-Shun; Li, Chuan-Feng; Guo, Guang-Can

2017-12-01

Quantum coherence is the most essential characteristic of quantum physics, specifcially, when it is subject to the resource-theoretical framework, it is considered as the most fundamental resource for quantum techniques. Other quantum resources, e.g., entanglement, are all based on coherence. Therefore, it becomes urgently important to learn how to preserve coherence in quantum channels. The best preservation is coherence freezing, which has been studied recently. However, in these studies, the freezing condition is theoretically calculated, and there still lacks a practical way to achieve this freezing; in addition the channels are usually fixed, but actually, there are also degrees of freedom that can be used to adapt the channels to quantum states. Here we develop a self-guided quantum coherence freezing method, which can guide either the quantum channels (tunable-channel scheme with upgraded channels) or the initial state (fixed-channel scheme) to the coherence-freezing zone from any starting estimate. Specifically, in the fixed-channel scheme, the final-iterative quantum states all satisfy the previously calculated freezing condition. This coincidence demonstrates the validity of our method. Our work will be helpful for the better protection of quantum coherence.

Effects of high pressure freezing (HPF) on denaturation of natural actomyosin extracted from prawn (Metapenaeus ensis).

PubMed

Cheng, Lina; Sun, Da-Wen; Zhu, Zhiwei; Zhang, Zhihang

2017-08-15

Effects of protein denaturation caused by high pressure freezing, involving Pressure-Factors (pressure, time) and Freezing-Factors (temperature, phase transition, recrystallization, ice crystal types), are complicated. In the current study, the conformation and functional changes of natural actomyosin (NAM) under pressure assisted freezing (PAF, 100,150,300,400,500MPa P -20°C/25min ), pressure shift freezing (PSF, 200MPa P -20°C/25min ), and immersion freezing ( 0.1MPa P -20°C/5min ) after pressure was released to 0.1MPa, as compared to normal immersion freezing process (IF, 0.1MPa P -20°C/30min ). Results indicated that PSF ( 200MPa P -20°C/30min ) could reduce the denaturation of frozen NAM and a pressure of 300MPa was the critical point to induce such a denaturation. During the periods of B→D in PSF or B→C→D in PAF, the generation and growth of ice crystals played an important role on changing the secondary and tertiary structure of the treated NAM. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of ultrasound-assisted freezing on the physico-chemical properties and volatile compounds of red radish.

PubMed

Xu, Bao-Guo; Zhang, Min; Bhandari, Bhesh; Cheng, Xin-Feng; Islam, Md Nahidul

2015-11-01

Power ultrasound, which can enhance nucleation rate and crystal growth rate, can also affect the physico-chemical properties of immersion frozen products. In this study, the influence of slow freezing (SF), immersion freezing (IF) and ultrasound-assisted freezing (UAF) on physico-chemical properties and volatile compounds of red radish was investigated. Results showed that ultrasound application significantly improved the freezing rate; the freezing time of ultrasound application at 0.26 W/cm(2) was shorten by 14% and 90%, compared to IF and SF, respectively. UAF products showed significant (p<0.05) reduction in drip loss and phytonutrients (anthocyanins, vitamin C and phenolics) loss. Compared to SF products, IF and UAF products showed better textural preservation and higher calcium content. The radish tissues exhibited better cellular structures under ultrasonic power intensities of 0.17 and 0.26 W/cm(2) with less cell separation and disruption. Volatile compound data revealed that radish aromatic profile was also affected in the freezing process. Copyright © 2015 Elsevier B.V. All rights reserved.

A comparative study of sodium dodecyl sulfate and freezing/thawing treatment on wheat starch: The role of water absorption.

PubMed

Tao, Han; Wang, Pei; Zhang, Bao; Wu, Fengfeng; Jin, Zhengyu; Xu, Xueming

2016-06-05

The effect of freezing on functionality of native and sodium dodecyl sulfate (SDS)-treated wheat starches was investigated, with the aim of understanding the role of water absorption during freezing process. SDS is one of most efficient detergents to remove non-starch components (such as proteins and lipids) for starches but does not cause any apparent damage on granular structure. Slow swelling could be converted to rapid swelling by SDS washing, indicating higher water absorption. Freezing process induced slight roughness on starch granules but the non-starch components content was little affected. Combined SDS+freezing treatment significantly decreased both amylose and proteins non-starch components contents, which was accompanied with high gelatinization temperatures, melting enthalpy, and pasting viscosities. A smaller bread specific volume was obtained from SDS+freezing-treated starches while the crumb firmness significantly increased (p<0.05). SDS mainly extracted the surface components from starch granules, leading to high water absorption and making granules sensitive to the freezing treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

A Vivens Ex Vivo Study on the Synergistic Effect of Electrolysis and Freezing on the Cell Nucleus

PubMed Central

Lugnani, Franco; Zanconati, Fabrizio; Marcuzzo, Thomas; Bottin, Cristina; Mikus, Paul; Guenther, Enric; Klein, Nina; Rubinsky, Liel; Stehling, Michael K.; Rubinsky, Boris

2015-01-01

Freezing—cryosurgery, and electrolysis—electrochemical therapy (EChT), are two important minimally invasive surgery tissue ablation technologies. Despite major advantages they also have some disadvantages. Cryosurgery cannot induce cell death at high subzero freezing temperatures and requires multiple freeze thaw cycles, while EChT requires high concentrations of electrolytic products—which makes it a lengthy procedure. Based on the observation that freezing increases the concentration of solutes (including products of electrolysis) in the frozen region and permeabilizes the cell membrane to these products, this study examines the hypothesis that there could be a synergistic effect between freezing and electrolysis in their use together for tissue ablation. Using an animal model we refer to as vivens ex vivo, which may be of value in reducing the use of animals for experiments, combined with a Hematoxylin stain of the nucleus, we show that there are clinically relevant protocols in which the cell nucleus appears intact when electrolysis and freezing are used separately but is affected by certain combinations of electrolysis and freezing. PMID:26695185

Potential weathering by freeze-thaw action in alpine rocks in the European Alps during a nine year monitoring period

NASA Astrophysics Data System (ADS)

Kellerer-Pirklbauer, Andreas

2017-11-01

A quantification of rock weathering by freeze-thaw processes in alpine rocks requires at least rock temperature data in high temporal resolution, in high quality, and over a sufficient period of time. In this study up to nine years of rock temperature data (2006-2015) from eleven rock monitoring sites in two of the highest mountain ranges of Austria were analyzed. Data were recorded at a half-hourly or hourly logging interval and at rock depths of 3, 10, and 30-40 cm. These data have been used to quantify mean conditions, ranges, and relationships of the potential near-surface weathering by freeze-thaw action considering volumetric-expansion of ice and ice segregation. For the former, freeze-thaw cycles and effective freeze-thaw cycles for frost shattering have been considered. For the latter, the intensity and duration of freezing events as well as time within the 'frost cracking window' have been analyzed. Results show that the eleven sites are in rather extreme topoclimatic positions and hence represent some of the highest and coolest parts of Austria and therefore the Eastern Alps. Only four sites are presumably affected by permafrost. Most sites are influenced by a long-lasting seasonal snow cover. Freeze-thaw cycles and effective freeze-thaw cycles for frost shattering are mainly affecting the near-surface and are unimportant at few tens of centimeters below the rock surface. The lowest temperatures during freezing events and the shortest freezing events have been quantified at all eleven monitoring sites very close to the surface. The time within the frost cracking window decreases in most cases from the rock surface inwards apart from very cold years/sites with very low temperatures close to the surface. As shown by this study and predicted climate change scenarios, assumed warmer rock temperature conditions in the future at alpine rock walls in Austria will lead to less severe freezing events and to shorter time periods within the frost-cracking window. Statistical correlation analyses showed furthermore that the longer the duration of the seasonal snow cover, the fewer are freeze-thaw cycles, the fewer are effective freeze-thaw cycles, the longer is the mean and the maximum duration of freezing events, and the lower is the mean annual ground temperature. The interaction of the winter snow cover history and the winter thermal regime has a complex effect on the duration of the frost cracking window but also on the number of freeze-thaw cycles as shown by a conceptual model. Predicted future warmer and snow-depleted winters in the European Alps will therefore have a complex impact on the potential weathering of alpine rocks by frost action which makes potential weathering predictions difficult. Neglecting rock moisture and rock properties in determining rock weathering limits the usefulness of solely rock temperature data. However, rock temperature data allow getting an estimate about potential weathering by freeze-thaw action which is often substantially more than previously known.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Allen, M.F.

A freeze-thaw power cycle is described that includes a piston driven by the expansion power of a fluid such as water in a cyclinder on freezing and the thawing thereof with alternate, rapid freezing and thawing of the fluid by low and high temperature means with heat transfer rates facilitated by the use of heat pipes or tubes or other conductor means to produce a continuous single or multi cyclinder quick, polution free and noiseless freeze/thaw power cycle.

References on Compression of Freeze-Dried Foods

DTIC Science & Technology

1978-08-01

ready-to-eat, freeze-dried scrambled egg . Swift and Co., Contract No. DA 19- 129-AMC-121. 67-49-FD (FD-54). January 1967 (AD 650637). In the design...significantly poorer in organoleptic properties than from grades A and B eggs. Freeze- dried scrambled egg packed in cans kept better in storage than when...scrambled, cooked, freeze-dried whole egg product was developed which possessed the appearance, aroma, flavor and texture similar to pan- fried scrambled

Method of quantifying the loss of acidification activity of lactic acid starters during freezing and frozen storage.

PubMed

Fonseca, F; Béal, C; Corrieu, G

2000-02-01

We have developed a method to quantify the resistance to freezing and frozen storage of lactic acid starters, based on measuring the time necessary to reach the maximum acidification rate in milk (tm) using the Cinac system. Depending on the operating conditions, tm increased during the freezing step and storage. The loss of acidification activity during freezing was quantified by the difference (delta tm) between the tm values of the concentrated cell suspension before and after freezing. During storage at -20 degrees C, linear relationships between tm and the storage time were established. Their slope, k, allowed the quantitation of the decrease in acidification activity during 9-14 weeks of frozen storage. The method was applied to determine the resistance to freezing and frozen storage of four strains of lactic acid bacteria and to quantify the cryoprotective effect of glycerol.

Influence of freezing down to 77.15 K on structure and antioxidant power of some proteins

NASA Astrophysics Data System (ADS)

Rozanova, S. L.; Narozhnyi, S. V.; Nardid, O. A.

2017-06-01

The purpose of the present work was to investigate influence of different freeze-thawing protocols on structure and antioxidant properties of isolated proteins. In our experiments we have studied human serum albumin, human hemoglobin and cytochrome C derived from equine heart frozen down to 77.15 K with 1-2 deg/min and 300 deg/min rate with following thawing on a water bath at 293.15 K. Native proteins were assumed as a control. Influence of freeze-thawing protocols on protein structure was investigated using spectrophotometric and fluorescent assays. Antioxidant activities of isolated proteins were estimated by their ability to reduce ABTS+ radical. It has been established that unfolding derived from freeze-thawing exposure leads to protein antioxidant activity increasing while decreasing of such an activity may be connected with macromolecule aggregation. Character of freeze-thawing influence on antioxidant activity of proteins depends on molecule structure peculiarities and freezing protocols.

Morphology of supercooled droplets freezing on solid surfaces

NASA Astrophysics Data System (ADS)

La, Shiren; Huang, Zhiting; Liu, Cong; Zhang, Xingyi

2018-05-01

Supercooled droplets freezing on solid surfaces are ubiquitous in nature. This letter investigates the influences of droplet viscosity on freezing velocity and frosting formation. Several experiments were conducted for three kinds of sessile droplets (water, silicone oil and oil) on two types of substrates (copper and iron) with different surface roughness at various temperatures. The results show that the water droplets exhibit obvious phase transition lines and their freezing speeds increase when the temperature of substrates decreases. It is found that the freezing speed is independent of the thermal conductivities of the substrates. Notably, the water droplets develop prominent bulges after freezing and subsequently nucleate to frost. In contrast, the high viscosity oil and silicone oil do not manifest an obvious phase transition line. Besides, no bulges are observed in these two kinds of droplets, suggesting that these frosting forms are of different mechanisms compared with water droplets.

Natural genetic variation of freezing tolerance in Arabidopsis.

PubMed

Hannah, Matthew A; Wiese, Dana; Freund, Susanne; Fiehn, Oliver; Heyer, Arnd G; Hincha, Dirk K

2006-09-01

Low temperature is a primary determinant of plant growth and survival. Using accessions of Arabidopsis (Arabidopsis thaliana) originating from Scandinavia to the Cape Verde Islands, we show that freezing tolerance of natural accessions correlates with habitat winter temperatures, identifying low temperature as an important selective pressure for Arabidopsis. Combined metabolite and transcript profiling show that during cold exposure, global changes of transcripts, but not of metabolites, correlate with the ability of Arabidopsis to cold acclimate. There are, however, metabolites and transcripts, including several transcription factors, that correlate with freezing tolerance, indicating regulatory pathways that may be of primary importance for this trait. These data identify that enhanced freezing tolerance is associated with the down-regulation of photosynthesis and hormonal responses and the induction of flavonoid metabolism, provide evidence for naturally increased nonacclimated freezing tolerance due to the constitutive activation of the C-repeat binding factors pathway, and identify candidate transcriptional regulators that correlate with freezing tolerance.

The effect of freeze-thawing on magnetic resonance imaging T2* of freshly harvested bovine patellar tendon

PubMed Central

Pownder, Sarah L.; Shah, Parina H.; Potter, Hollis G.

2015-01-01

Background Analysis of fresh specimens in research studies is ideal; however, it is often necessary to freeze samples for evaluation at a later time. Limited evaluation of the effect of freeze-thawing of tendon tissue samples on inherent magnetic resonance imaging (MRI) parameters, such as ultrashort echo time (UTE) T2* values, have been performed to date. Methods This study performed UTE MRI on 14 bovine patellar tendons at harvest and after four consecutive freeze-thaw cycles. Results Results demonstrated a small but significant reduction (12%) in tendon T2* values after the first freeze thaw cycle, but not after successive cycles. Tendons from juvenile animals with open physis had a significant reduction of T2* following a single freeze thaw cycle, P<0.0001. Conclusions The results of this study emphasize the importance of using uniform tendon storage protocols when using UTE MRI in preclinical models. PMID:26029639

Limited Impact of Subglacial Supercooling Freeze-on for Greenland Ice Sheet Stratigraphy

NASA Astrophysics Data System (ADS)

Dow, Christine F.; Karlsson, Nanna B.; Werder, Mauro A.

2018-02-01

Large units of disrupted radiostratigraphy (UDR) are visible in many radio-echo sounding data sets from the Greenland Ice Sheet. This study investigates whether supercooling freeze-on rates at the bed can cause the observed UDR. We use a subglacial hydrology model to calculate both freezing and melting rates at the base of the ice sheet in a distributed sheet and within basal channels. We find that while supercooling freeze-on is a phenomenon that occurs in many areas of the ice sheet, there is no discernible correlation with the occurrence of UDR. The supercooling freeze-on rates are so low that it would require tens of thousands of years with minimal downstream ice motion to form the hundreds of meters of disrupted radiostratigraphy. Overall, the melt rates at the base of the ice sheet greatly overwhelm the freeze-on rates, which has implications for mass balance calculations of Greenland ice.

Realization of the Temperature Scale in the Range from 234.3 K (Hg Triple Point) to 1084.62°C (Cu Freezing Point) in Croatia

NASA Astrophysics Data System (ADS)

Zvizdic, Davor; Veliki, Tomislav; Grgec Bermanec, Lovorka

2008-06-01

This article describes the realization of the International Temperature Scale in the range from 234.3 K (mercury triple point) to 1084.62°C (copper freezing point) at the Laboratory for Process Measurement (LPM), Faculty of Mechanical Engineering and Naval Architecture (FSB), University of Zagreb. The system for the realization of the ITS-90 consists of the sealed fixed-point cells (mercury triple point, water triple point and gallium melting point) and the apparatus designed for the optimal realization of open fixed-point cells which include the gallium melting point, tin freezing point, zinc freezing point, aluminum freezing point, and copper freezing point. The maintenance of the open fixed-point cells is described, including the system for filling the cells with pure argon and for maintaining the pressure during the realization.

The principles of ultrasound and its application in freezing related processes of food materials: A review.

PubMed

Cheng, Xinfeng; Zhang, Min; Xu, Baoguo; Adhikari, Benu; Sun, Jincai

2015-11-01

Ultrasonic processing is a novel and promising technology in food industry. The propagation of ultrasound in a medium generates various physical and chemical effects and these effects have been harnessed to improve the efficiency of various food processing operations. Ultrasound has also been used in food quality control as diagnostic technology. This article provides an overview of recent developments related to the application of ultrasound in low temperature and closely related processes such as freezing, thawing, freeze concentration and freeze drying. The applications of high intensity ultrasound to improve the efficiency of freezing process, to control the size and size distribution of ice crystals and to improve the quality of frozen foods have been discussed in considerable detail. The use of low intensity ultrasound in monitoring the ice content and to monitor the progress of freezing process has also been highlighted. Copyright © 2015 Elsevier B.V. All rights reserved.

Freeze-drying process design by manometric temperature measurement: design of a smart freeze-dryer.

PubMed

Tang, Xiaolin Charlie; Nail, Steven L; Pikal, Michael J

2005-04-01

To develop a procedure based on manometric temperature measurement (MTM) and an expert system for good practices in freeze drying that will allow development of an optimized freeze-drying process during a single laboratory freeze-drying experiment. Freeze drying was performed with a FTS Dura-Stop/Dura-Top freeze dryer with the manometric temperature measurement software installed. Five percent solutions of glycine, sucrose, or mannitol with 2 ml to 4 ml fill in 5 ml vials were used, with all vials loaded on one shelf. Details of freezing, optimization of chamber pressure, target product temperature, and some aspects of secondary drying are determined by the expert system algorithms. MTM measurements were used to select the optimum shelf temperature, to determine drying end points, and to evaluate residual moisture content in real-time. MTM measurements were made at 1 hour or half-hour intervals during primary drying and secondary drying, with a data collection frequency of 4 points per second. The improved MTM equations were fit to pressure-time data generated by the MTM procedure using Microcal Origin software to obtain product temperature and dry layer resistance. Using heat and mass transfer theory, the MTM results were used to evaluate mass and heat transfer rates and to estimate the shelf temperature required to maintain the target product temperature. MTM product dry layer resistance is accurate until about two-thirds of total primary drying time is over, and the MTM product temperature is normally accurate almost to the end of primary drying provided that effective thermal shielding is used in the freeze-drying process. The primary drying times can be accurately estimated from mass transfer rates calculated very early in the run, and we find the target product temperature can be achieved and maintained with only a few adjustments of shelf temperature. The freeze-dryer overload conditions can be estimated by calculation of heat/mass flow at the target product temperature. It was found that the MTM results serve as an excellent indicator of the end point of primary drying. Further, we find that the rate of water desorption during secondary drying may be accurately measured by a variation of the basic MTM procedure. Thus, both the end point of secondary drying and real-time residual moisture may be obtained during secondary drying. Manometric temperature measurement and the expert system for good practices in freeze drying does allow development of an optimized freeze-drying process during a single laboratory freeze-drying experiment.

Freezing-induced deformation of biomaterials in cryomedicine

NASA Astrophysics Data System (ADS)

Ozcelikkale, Altug

Cryomedicine utilizes low temperature treatments of biological proteins, cells and tissues for cryopreservation, materials processing and cryotherapy. Lack of proper understanding of cryodamage that occurs during these applications remains to be the primary bottleneck for development of successful tissue cryopreservation and cryosurgery procedures. An engineering approach based on a view of biological systems as functional biomaterials can help identify, predict and control the primary cryodamage mechanisms by developing an understanding of underlying freezing-induced biophysical processes. In particular, freezing constitutes the main structural/mechanical origin of cryodamage and results in significant deformation of biomaterials at multiple length scales. Understanding of these freezing-induced deformation processes and their effects on post-thaw biomaterial functionality is currently lacking but will be critical to engineer improved cryomedicine procedures. This dissertation addresses this problem by presenting three separate but related studies of freezing-induced deformation at multiple length scales including nanometer-scale protein fibrils, single cells and whole tissues. A combination of rigorous experimentation and computational modeling is used to characterize post-thaw biomaterial structure and properties, predict biomaterial behavior and assess its post-thaw biological functionality. Firstly, freezing-induced damage on hierarchical extracellular matrix structure of collagen is investigated at molecular, fibril and matrix levels. Results indicate to a specific kind of fibril damage due to freezing-induced expansion of intrafibrillar fluid. This is followed by a study of freezing-induced cell and tissue deformation coupled to osmotically driven cellular water transport. Computational and semi empirical modeling of these processes indicate that intracellular deformation of the cell during freezing is heterogeneous and can interfere with cellular water transport, thereby leading to previously unconsidered mechanisms of cell freezing response. In addition, cellular water transport is identified as the critical limiting factor on the amount of freezing-induced tissue deformation, particularly in native tissues with high cell densities. Finally, effects of cryopreservation on post-thaw biological functionality of collagen engineered tissue constructs is investigated where cell-matrix interactions during fibroblast migration are considered as the functional response. Simultaneous cell migration and extracellular matrix deformation are characterized. Results show diminished cell-matrix coupling by freeze/thaw accompanied by a subtle decrease in cell migration. A connection between these results and freezing-induced collagen fibril damage is also suggested. Overall, this dissertation provides new fundamental knowledge on cryodamage mechanisms and a collection of novel multi-purpose engineering tools that will open the way for rational design of cryomedicine technologies.

An Equipment to Measure the Freezing Point of Soils under Higher Pressure

NASA Astrophysics Data System (ADS)

Wang, Dayan; Guan, Hui; Wen, Zhi; Ma, Wei

2014-05-01

Soil freezing point is the highest temperature at which ice can be presented in the system and soil can be referred to as frozen. The freezing temperature of soil is an important parameter for solving many practical problems in civil engineering, such as evaluation of soil freezing depth, prediction of soil heaving, force of soil suction, etc. However, as the freezing temperature is always affected by many factors like soil particle size, mineral composition, water content and the external pressure endured by soils, to measure soil freezing point is a rather difficult task until now, not to mention the soil suffering higher pressure. But recently, with the artificial freezing technology widely used in the excavation of deep underground space, the frozen wall thickness is a key factor to impact the security and stability of deep frozen wall. To determine the freeze wall thickness, the location of the freezing front must be determined firstly, which will deal with the determination of the soil freezing temperature. So how to measure the freezing temperature of soil suffering higher pressure is an important problem to be solved. This paper will introduce an equipment which was developed lately by State Key Laboratory of Frozen Soil Engineering to measure the freezing-point of soils under higher pressure. The equipment is consisted of cooling and keeping temperature system, temperature sensor and data collection system. By cooling and keeping temperature system, not only can we make the higher pressure soil sample's temperature drop to a discretionary minus temperature, but also keep it and reduce the heat exchange of soil sample with the outside. The temperature sensor is the key part to our measurement, which is featured by high precision and high sensitivity, what is more important is that the temperature sensor can work in a higher pressure condition. Moreover, the major benefit of this equipment is that the soil specimen's loads can be loaded by any microcomputer control electron universal testing machines. All of above mentioned advantages of this equipment ensures one to catch up the moment soil turns from the thawed state into ice and enable one to determine the freezing point experimentally by recording the temperature-time history (cooling curve) at particular points within the sample used for analysis. Therefore, this equipment has excellent characteristics such as compact construction, convenient operation, high reliability and the measuring accuracy. The authors would like to thank the following agents for their financial supports: the National Natural Science Foundation (No.41071048),Hundred Talent Young Scientists program of the Chinese Academy of Sciences granted to Dr. Zhi Wen.

Evaluation of a New Motion-correction Algorithm Using On-rigid Registration in Respiratory-gated PET/CT Images of Liver Tumors.

PubMed

Wagatsuma, Kei; Osawa, Tatsufumi; Yokokawa, Naoki; Miwa, Kenta; Oda, Keiichi; Kudo, Yoshiro; Unno, Yasushi; Ito, Kimiteru; Ishii, Kenji

2016-01-01

The present study aimed to determine the qualitative and quantitative accuracy of the Q.Freeze algorithm in PET/CT images of liver tumors. A body phantom and hot spheres representing liver tumors contained 5.3 and 21.2 kBq/mL of a solution containing 18 F radioactivity, respectively. The phantoms were moved in the superior-inferior direction at a motion displacement of 20 mm. Conventional respiratory-gated (RG) and Q.Freeze images were sorted into 6, 10, and 13 phase-groups. The SUV ave was calculated from the background of the body phantom, and the SUV max was determined from the hot spheres of the liver tumors. Three patients with four liver tumors were also clinically assessed by whole-body and RG PET. The RG and Q.Freeze images derived from the clinical study were also sorted into 6, 10 and 13 phase-groups. Liver signal-to-noise ratio (SNR) and SUV max were determined from the RG and Q.Freeze clinical images. The SUV ave of Q.Freeze images was the same as those derived from the body phantom using RG. The liver SNR improved with Q.Freeze, and the SUVs max was not overestimated when Q.Freeze was applied in both the phantom and clinical studies. Q.Freeze did not degrade the liver SNR and SUV max even though the phase number was larger. Q.Freeze delivered qualitative and quantitative motion correction than conventional RG imaging even in 10-phase groups.

A numerical model for water and heat transport in freezing soils with nonequilibrium ice-water interfaces

NASA Astrophysics Data System (ADS)

Peng, Zhenyang; Tian, Fuqiang; Wu, Jingwei; Huang, Jiesheng; Hu, Hongchang; Darnault, Christophe J. G.

2016-09-01

A one-dimensional numerical model of heat and water transport in freezing soils is developed by assuming that ice-water interfaces are not necessarily in equilibrium. The Clapeyron equation, which is derived from a static ice-water interface using the thermal equilibrium theory, cannot be readily applied to a dynamic system, such as freezing soils. Therefore, we handled the redistribution of liquid water with the Richard's equation. In this application, the sink term is replaced by the freezing rate of pore water, which is proportional to the extent of supercooling and available water content for freezing by a coefficient, β. Three short-term laboratory column simulations show reasonable agreement with observations, with standard error of simulation on water content ranging between 0.007 and 0.011 cm3 cm-3, showing improved accuracy over other models that assume equilibrium ice-water interfaces. Simulation results suggest that when the freezing front is fixed at a specific depth, deviation of the ice-water interface from equilibrium, at this location, will increase with time. However, this deviation tends to weaken when the freezing front slowly penetrates to a greater depth, accompanied with thinner soils of significant deviation. The coefficient, β, plays an important role in the simulation of heat and water transport. A smaller β results in a larger deviation in the ice-water interface from equilibrium, and backward estimation of the freezing front. It also leads to an underestimation of water content in soils that were previously frozen by a rapid freezing rate, and an overestimation of water content in the rest of the soils.

Genome-Wide Investigation of MicroRNAs and Their Targets in Response to Freezing Stress in Medicago sativa L., Based on High-Throughput Sequencing

PubMed Central

Shu, Yongjun; Liu, Ying; Li, Wei; Song, Lili; Zhang, Jun; Guo, Changhong

2016-01-01

Winter damage, especially in northern climates, is a major limitation of the utilization of perennial forages such as alfalfa. Therefore, improving freezing tolerance is imperative in alfalfa genetic breeding. However, freezing tolerance is a complex trait that is determined by many genes. To understand the complex regulation mechanisms of freezing tolerance in alfalfa, we performed small RNA sequencing analysis under cold (4°) and freezing (−8°) stress. The sequencing results revealed that 173 known, and 24 novel miRNAs were expressed, and that the expression of 35 miRNAs was affected by cold and/or freezing stress. Meanwhile, 105 target genes cleaved by these miRNAs were characterized by degradome sequencing. These targets were associated with biological regulation, cellular processes, metabolic processes, and response to stress. Interestingly, most of them were characterized as transcription factors (TFs), including auxin response factors, SBP, NAC, AP2/ERF, and GRF, which play important roles in plant abiotic responses. In addition, important miRNAs and mRNAs involved in nodulation were also identified, for example, the relationship between miR169 and the TF CCAAT (also named as NF-YA/HAP2), which suggested that nodulation has an important function in freezing tolerance in alfalfa. Our results provide valuable information to help determine the molecular mechanisms of freezing tolerance in alfalfa, which will aid the application of these miRNAs and their targets in the improvement of freezing tolerance in alfalfa and related plants. PMID:26801649

Heat and mass transfer scale-up issues during freeze-drying, III: control and characterization of dryer differences via operational qualification tests.

PubMed

Rambhatla, S; Tchessalov, S; Pikal, Michael J

2006-04-21

The objective of this research was to estimate differences in heat and mass transfer between freeze dryers due to inherent design characteristics using data obtained from sublimation tests. This study also aimed to provide guidelines for convenient scale-up of the freeze-drying process. Data obtained from sublimation tests performed on laboratory-scale, pilot, and production freeze dryers were used to evaluate various heat and mass transfer parameters: nonuniformity in shelf surface temperatures, resistance of pipe, refrigeration system, and condenser. Emissivity measurements of relevant surfaces such as the chamber wall and the freeze dryer door were taken to evaluate the impact of atypical radiation heat transfer during scale-up. "Hot" and "cold" spots were identified on the shelf surface of different freeze dryers, and the impact of variation in shelf surface temperatures on the primary drying time and the product temperature during primary drying was studied. Calculations performed using emissivity measurements on different freeze dryers suggest that a front vial in the laboratory lyophilizer received 1.8 times more heat than a front vial in a manufacturing freeze dryer operating at a shelf temperature of -25 degrees C and a chamber pressure of 150 mTorr during primary drying. Therefore, front vials in the laboratory are much more atypical than front vials in manufacturing. Steady-state heat and mass transfer equations were used to study a combination of different scale-up issues pertinent during lyophilization cycles commonly used for the freeze-drying of pharmaceuticals.

Cold tolerance of Littorinidae from southern Africa: intertidal snails are not constrained to freeze tolerance.

PubMed

Sinclair, Brent J; Marshall, David J; Singh, Sarika; Chown, Steven L

2004-11-01

All intertidal gastropods for which cold tolerance strategies have been assessed have been shown to be freeze tolerant. Thus, freeze tolerance is considered an adaptation to the intertidal environment. We investigated the cold tolerance strategies of three species of subtropical and temperate snails (Gastropoda: Littorinidae) to determine whether this group is phylogenetically constrained to freeze tolerance. We exposed 'dry' acclimated and 'wet' rehydrated snails to low temperatures to determine temperature of crystallisation (Tc), lower lethal temperature and LT(50) and to examine the relationship between ice formation and mortality. Tc was lowest in dry Afrolittorina knysnaensis (-13.6+/-0.4 degrees C), followed by dry Echinolittorina natalensis (-10.9+/-0.2 degrees C) and wet A. knysnaensis (-10.2+/-0.2 degrees C) . The Tc of both A. knysnaensis and E. natalensis increased with rehydration, whereas Tc of dry and wet Afrolittorina africana did not differ (-9.6+/-0.2 and -9.0+/-0.2 degrees C respectively). Wet snails of all species exhibited no or low survival of inoculative freezing, whereas dry individuals of A. knysnaensis could survive subzero temperatures above -8 degrees C when freezing was inoculated with ice . In the absence of external ice, Afrolittorina knysnaensis employs a freeze-avoidance strategy of cold tolerance, the first time this has been reported for an intertidal snail, indicating that there is no family-level phylogenetic constraint to freeze tolerance. Echinolittorina natalensis and A. africana both showed pre-freeze mortality and survival of some internal ice formation, but were not cold hardy in any strict sense.

Effects of rasagiline on freezing of gait in Parkinson's disease - an open-label, multicenter study.

PubMed

Cibulcik, Frantisek; Benetin, Jan; Kurca, Egon; Grofik, Milan; Dvorak, Miloslav; Richter, Denis; Donath, Vladimir; Kothaj, Jan; Minar, Michal; Valkovic, Peter

2016-12-01

Freezing of gait is a disabling symptom in advanced Parkinson's disease. Positive effects have been suggested with MAO-B inhibitors. We report on an open label clinical study on the efficacy of rasagiline as add-on therapy on freezing of gait and quality of life in patients with Parkinson's disease. Forty two patients with freezing of gait were treated with 1 mg rasagiline daily as an add-on therapy. Patients were assessed at baseline and after 1, 2 and 3 months of treatment. Freezing of gait severity was assessed using the Freezing of Gait Questionnaire, motor impairment by the modified MDS UPDRS part III, and quality of life using the PDQ-39 questionnaire. Patients treated with rasagiline had a statistically significant decrease in FoG-Q score and modified MDS UPDRS score after 1, 2 and 3 months of therapy. A moderately strong (r = 0.686, P = 0.002) correlation between the effects on mobility and freezing of gait was found. We also observed a statistically significant improvement in global QoL and in the subscales mobility, ADL, stigma and bodily discomfort in patients after 3 months of rasagiline therapy. A significant correlation (r = 0.570, P = 0.02) between baseline FoG-Q score and the baseline score for the PDQ Mobility subscale was found. In our study rasagiline as add-on antiparkinsonian therapy significantly improved mobility, freezing of gait and quality of life. The positive effect on freezing of gait appears to be related to improvement of mobility.

The Impact of Repeated Freeze-Thaw Cycles on the Quality of Biomolecules in Four Different Tissues.

PubMed

Ji, Xiaoli; Wang, Min; Li, Lingling; Chen, Fang; Zhang, Yanyang; Li, Qian; Zhou, Junmei

2017-10-01

High-quality biosamples are valuable resources for biomedical research. However, some tissues are stored without being sectioned into small aliquots and have to undergo repeated freeze-thaw cycles throughout prolonged experimentation. Little is known regarding the effects of repeated freeze-thaw cycles on the quality of biomolecules in tissues. The aim of this study was to evaluate the impact of repeated freeze-thaw (at room temperature or on ice) cycles on biomolecules and gene expression in four different types of tissues. Each fresh tissue was sectioned into seven aliquots and snap-frozen before undergoing repeated freeze-thaw cycles at room temperature or on ice. Biomolecules were extracted and analyzed. Both relative and absolute quantification were used to detect the changes in gene expression. The results indicated that the impact of repeated freeze-thaw cycles on RNA integrity varied by tissue type. Gene expression, including the housekeeping gene, was affected in RNA-degraded samples according to absolute quantification rather than relative quantification. Furthermore, our results suggest that thawing on ice could protect RNA integrity compared with thawing at room temperature. No obvious degradation of protein or DNA was observed with repeated freeze-thaw cycles either at room temperature or on ice. This research provides ample evidence for the necessity of sectioning fresh tissues into small aliquots before snap-freezing, thus avoiding degradation of RNA and alteration of gene expression resulting from repeated freeze-thaw cycles. For frozen tissue samples that were already in storage and had to be used repeatedly during their lifecycle, thawing on ice or sectioned at ultralow temperature is recommended.

The human milk oligosaccharides are not affected by pasteurization and freeze-drying.

PubMed

Hahn, Won-Ho; Kim, Jaehan; Song, Seunghyun; Park, Suyeon; Kang, Nam Mi

2017-11-06

Human milk oligosaccharides (HMOs) are known as important factors in neurologic and immunologic development of neonates. Moreover, freeze-drying seems to be a promising storage method to improve the processes of human milk banks. However, the effects of pasteurization and freeze-drying on HMOs were not evaluated yet. The purpose of this study is to analyze and compare the HMOs profiles of human milk collected before and after the pasteurization and freeze-drying. Totally nine fresh human milk samples were collected from three healthy mothers at the first, second, and third week after delivery. The samples were treated with Holder pasteurization and freeze-drying. HMOs profiles were analyzed by matrix-assisted laser desorption/ionization (MALDI) time-of-flight/time-of-flight (TOF/TOF) mass spectrometry and compared between samples collected before and after the treatments. Human milk samples showed significantly different HMO patterns between mothers. However, HMOs were not affected by lactation periods within 3 weeks after delivery (r 2  = 0.972-0.999, p < .001). Moreover, both of pasteurization and freeze-drying were found not to affect HMO patterns in a correlation analysis (r 2  = 0.989-0.999, p < .001). HMO patterns were found not to be affected by pasteurization and freeze-drying of donor milks. We hope that introducing freeze-drying to the human milk banks would be encouraged by the present study. However, the storage length without composition changes of HMOs after freeze-drying needs to be evaluated in the further studies.

The importance of heat flow direction for reproducible and homogeneous freezing of bulk protein solutions.

PubMed

Rodrigues, Miguel A; Balzan, Gustavo; Rosa, Mónica; Gomes, Diana; de Azevedo, Edmundo G; Singh, Satish K; Matos, Henrique A; Geraldes, Vítor

2013-01-01

Freezing is an important operation in biotherapeutics industry. However, water crystallization in solution, containing electrolytes, sugars and proteins, is difficult to control and usually leads to substantial spatial solute heterogeneity. Herein, we address the influence of the geometry of freezing direction (axial or radial) on the heterogeneity of the frozen matrix, in terms of local concentration of solutes and thermal history. Solutions of hemoglobin were frozen radially and axially using small-scale and pilot-scale freezing systems. Concentration of hemoglobin, sucrose and pH values were measured by ice-core sampling and temperature profiles were measured at several locations. The results showed that natural convection is the major source for the cryoconcentration heterogeneity of solutes over the geometry of the container. A significant improvement in this spatial heterogeneity was observed when the freezing geometry was nonconvective, i.e., the freezing front progression was unidirectional from bottom to top. Using this geometry, less than 10% variation in solutes concentration was obtained throughout the frozen solutions. This result was reproducible, even when the volume was increased by two orders of magnitude (from 30 mL to 3 L). The temperature profiles obtained for the nonconvective freezing geometry were predicted using a relatively simple computational fluid dynamics model. The reproducible solutes distribution, predictable temperature profiles, and scalability demonstrate that the bottom to top freezing geometry enables an extended control over the freezing process. This geometry has therefore shown the potential to contribute to a better understanding and control of the risks inherent to frozen storage. © 2013 American Institute of Chemical Engineers.

Water vapor movement in freezing aggregate base materials.

DOT National Transportation Integrated Search

2014-06-01

The objectives of this research were to 1) measure the extent to which water vapor movement results in : water accumulation in freezing base materials; 2) evaluate the effect of soil stabilization on water vapor movement : in freezing base materials;...

Freezing temperature protection admixture for Portland cement concrete

DOT National Transportation Integrated Search

1996-10-01

A number of experimental admixtures were compared to Pozzutec 20 admixture for their ability to protect fresh concrete from freezing and for increasing the rate of cement hydration at below-freezing temperatures. The commercial accelerator and low-te...

Freezing tolerance of conifer seeds and germinants.

PubMed

Hawkins, B J; Guest, H J; Kolotelo, D

2003-12-01

Survival after freezing was measured for seeds and germinants of four seedlots each of interior spruce (Picea glauca x engelmannii complex), lodgepole pine (Pinus contorta Dougl. ex Loud.), Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and western red cedar (Thuja plicata Donn ex D. Donn). Effects of eight seed treatments on post-freezing survival of seeds and germinants were tested: dry, imbibed and stratified seed, and seed placed in a growth chamber for 2, 5, 10, 15, 20 or 30 days in a 16-h photoperiod and a 22/17 degrees C thermoperiod. Survival was related to the water content of seeds and germinants, germination rate and seedlot origin. After freezing for 3 h at -196 degrees C, dry seed of most seedlots of interior spruce, Douglas-fir and western red cedar had 84-96% germination, whereas lodgepole pine seedlots had 53-82% germination. Freezing tolerance declined significantly after imbibition in lodgepole pine, Douglas-fir and interior spruce seed (western red cedar was not tested), and mean LT50 of imbibed seed of these species was -30, -24.5 and -20 degrees C, respectively. Freezing tolerance continued to decline to a minimum LT50 of -4 to -7 degrees C after 10 days in a growth chamber for interior spruce, Douglas-fir and lodgepole pine, or after 15 days for western red cedar. Minimum freezing tolerance was reached at the stage of rapid hypocotyl elongation. In all species, a slight increase in freezing tolerance of germinants was observed once cotyledons emerged from the seed coat. The decrease in freezing tolerance during the transition from dry to germinating seed correlated with increases in seed water content. Changes in freezing tolerance between 10 and 30 days in the growth chamber were not correlated with seedling water content. Within a species, seedlots differed significantly in freezing tolerance after 2 or 5 days in the growth chamber. Because all seedlots of interior spruce and lodgepole pine germinated quickly, there was no correlation between seedlot hardiness and rate of germination. Germination rate and freezing tolerance of Douglas-fir and western red cedar seedlots was negatively correlated. There was a significant correlation between LT50 after 10 days in the growth chamber and minimum spring temperature at the location of seedlot origin for interior spruce and three seedlots of western red cedar, but no relationship was apparent for lodgepole pine and Douglas-fir.

A comparative study of K-rich and Na/Ca-rich feldspar ice-nucleating particles in a nanoliter droplet freezing assay

NASA Astrophysics Data System (ADS)

Peckhaus, Andreas; Kiselev, Alexei; Hiron, Thibault; Ebert, Martin; Leisner, Thomas

2016-09-01

A recently designed droplet freezing assay was used to study the freezing of up to 1500 identical 0.2 nL water droplets containing suspensions of one Na/Ca-rich feldspar and three K-rich and one Na/Ca-rich feldspar particles. Three types of experiments have been conducted: cooling ramp, isothermal freezing at a constant temperature, and freeze-thaw cycles. The observed freezing behavior has been interpreted with the help of a model based on the classical nucleation theory (soccer ball model (SBM); Niedermeier et al., 2015). By applying the model to the different freezing experiments conducted with the same ice-nucleating material, the unique sets of model parameters for specific feldspar suspensions could be derived. The SBM was shown to adequately describe the observed cooling rate dependence, the ice-nucleating active sites (INAS) surface density ns(T) in a wide temperature range, and the shift of the freezing curves towards lower temperature with dilution. Moreover, the SBM was capable of reproducing the variation of INAS surface density ns(T) with concentration of ice-nucleating particles in the suspension droplets and correctly predicting the leveling-off of ns(T) at low temperature. The freeze-thaw experiments have clearly shown that the heterogeneous freezing induced even by very active ice-nucleating species still possesses a stochastic nature, with the degree of randomness increasing towards homogeneous nucleation. A population of the high-temperature INAS has been identified in one of the K-rich feldspar samples. The freezing of 0.8 wt % suspension droplets of this particular feldspar was observed already at -5 °C. These high-temperature active sites could be deactivated by treating the sample with hydrogen peroxide but survived heating up to 90 °C. Given a high mass concentration of these high-temperature active sites (2.9 × 108 g-1) and a very low value of contact angle (0.56 rad) the possibility of biological contamination of the sample was concluded to be unlikely but could not be completely ruled out. The freezing efficacy of all feldspar samples has been shown to reduce only slightly after suspension in water for over 5 months.

Storing Blood Cells

NASA Technical Reports Server (NTRS)

1976-01-01

The National Cancer Institute worked with Goddard Space Flight Center to propose a solution to the blood-cell freezing problem. White blood cells and bone marrow are stored for future use by leukemia patients as a result of Goddard and Jet Propulsion Laboratory expertise in electronics and cryogenics. White blood cell and bone marrow bank established using freezing unit. Freezing unit monitors temperature of cells themselves. Thermocouple placed against polyethylene container relays temperature signals to an electronic system which controls small heaters located outside container. Heaters allow liquid nitrogen to circulate at constant temperature and maintain consistent freezing rate. Ability to freeze, store, and thaw white cells and bone marrow without damage is important in leukemia treatment.

Effect of demedullation on freezing injury in hind limbs of rats

NASA Astrophysics Data System (ADS)

Dhingra, Shashi; Bhatia, B.; Chhina, G. S.; Singh, Baldev

1987-09-01

Freezing incidence and tissue loss on exposure of hind limbs of female Wistar rats to freezing mixture was reduced by demedullation 6 days prior to cold exposure (p<0.01 and p<0.001 respectively); demedullation 1 h after freezing injury had no effect on tissue loss. Noradrenaline (1 mg/kg i.p.) 5 min before exposure increased the freezing incidence in intact (p<0.05) as well as in demedullated rats (p<0.01), with no effect on tissue loss. Adrenaline (500 mg/kg i.p.) had no effect on either. A sustained fall in plasma adrenaline after demedullation leading to reduced reactivity of the blood vessels to some vasoactive agents is postulated.

Extending the Season for Concrete Construction and Repair. Phase II - Defining Engineering Parameters

DTIC Science & Technology

2006-04-01

dosages, seemed to improve the freeze –thaw durability of concrete. Phase II found what appears to be a maximum dosage after which freeze –thaw...durability becomes a concern. That is because cement hydration can only create a finite amount of space to absorb these chemicals. Thus, for freeze ...protection, admixture dosages should be designed according to water content as specified in Phase I, while, for freeze –thaw durability, admixture dosages

Freeze-Drying of Plant Tissue Containing HBV Surface Antigen for the Oral Vaccine against Hepatitis B

PubMed Central

Milczarek, Magdalena; Pajtasz-Piasecka, Elżbieta; Wietrzyk, Joanna

2014-01-01

The aim of this study was to develop a freeze-drying protocol facilitating successful processing of plant material containing the small surface antigen of hepatitis B virus (S-HBsAg) while preserving its VLP structure and immunogenicity. Freeze-drying of the antigen in lettuce leaf tissue, without any isolation or purification step, was investigated. Each process step was consecutively evaluated and the best parameters were applied. Several drying profiles and excipients were tested. The profile of 20°C for 20 h for primary and 22°C for 2 h for secondary drying as well as sucrose expressed efficient stabilisation of S-HBsAg during freeze-drying. Freezing rate and postprocess residual moisture were also analysed as important factors affecting S-HBsAg preservation. The process was reproducible and provided a product with VLP content up to 200 µg/g DW. Assays for VLPs and total antigen together with animal immunisation trials confirmed preservation of antigenicity and immunogenicity of S-HBsAg in freeze-dried powder. Long-term stability tests revealed that the stored freeze-dried product was stable at 4°C for one year, but degraded at elevated temperatures. As a result, a basis for an efficient freeze-drying process has been established and a suitable semiproduct for oral plant-derived vaccine against HBV was obtained. PMID:25371900

SLAPex Freeze/Thaw 2015: The First Dedicated Soil Freeze/Thaw Airborne Campaign

NASA Technical Reports Server (NTRS)

Kim, Edward; Wu, Albert; DeMarco, Eugenia; Powers, Jarrett; Berg, Aaron; Rowlandson, Tracy; Freeman, Jacqueline; Gottfried, Kurt; Toose, Peter; Roy, Alexandre;

Effect of photoperiod prior to cold acclimation on freezing tolerance and carbohydrate metabolism in alfalfa (Medicago sativa L.).

PubMed

Bertrand, Annick; Bipfubusa, Marie; Claessens, Annie; Rocher, Solen; Castonguay, Yves

2017-11-01

Cold acclimation proceeds sequentially in response to decreases in photoperiod and temperature. This study aimed at assessing the impact of photoperiod prior to cold acclimation on freezing tolerance and related biochemical and molecular responses in two alfalfa cultivars. The fall dormant cultivar Evolution and semi-dormant cultivar 6010 were grown in growth chambers under different photoperiods (8, 10, 12, 14 or 16h) prior to cold acclimation. Freezing tolerance was evaluated as well as carbohydrate concentrations, levels of transcripts encoding enzymes of carbohydrate metabolism as well as a K-3dehydrin, before and after cold acclimation. The fall dormant cultivar Evolution had a better freezing tolerance than the semi-dormant cultivar 6010. The effect of photoperiod prior to cold acclimation on the level of freezing tolerance differed between the two cultivars: an 8h-photoperiod induced the highest level of freezing tolerance in Evolution and the lowest in 6010. In Evolution, the 8h-induced superior freezing tolerance was associated with higher concentration of raffinose-family oligosaccharides (RFO). The transcript levels of sucrose synthase (SuSy) decreased whereas those of sucrose phosphatase synthase (SPS) and galactinol synthase (GaS) increased in response to cold acclimation in both cultivars. Our results indicate that RFO metabolism could be involved in short photoperiod-induced freezing tolerance in dormant alfalfa cultivars. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

Kinetic freeze-out conditions for the production of resonances, hadronic molecules, and light nuclei

NASA Astrophysics Data System (ADS)

Cho, Sungtae; Song, Taesoo; Lee, Su Houng

2018-02-01

We investigate the freeze-out conditions of a particle in an expanding system of interacting particles in order to understand the productions of resonances, hadronic molecules, and light nuclei in heavy-ion collisions. Applying the kinetic freeze-out condition with explicit hydrodynamic calculations for the expanding hadronic phase to the daughter particles of K* mesons, we find that the larger suppression of the yield ratio of K*/K at the Large Hadron Collider (LHC) than at the Relativisitic Heavy Ion Collider (RHIC) compared to the expectations from the statistical hadronization model based on chemical freeze-out parameters reflects the lower kinetic freeze-out temperature at LHC than at RHIC. Furthermore, we point out that for the light nuclei or hadronic molecules that are bound, the freeze-out condition should be applied to the respective particle in the hadronic matter. It is then shown through the rate equation that when the nucleon and pion numbers are kept constant at the chemical freeze-out value during the hadronic phase, the deuteron number quickly approaches an asymptotic value that is close to the statistical model prediction at the chemical freeze-out point. We argue that the reduction seen in K* numbers is a typical result for a particle that has a large natural decay width decaying into daughter particles, while that for deuteron is typical for a stable hadronic bound state.

Stem water transport and freeze-thaw xylem embolism in conifers and angiosperms in a Tasmanian treeline heath.

PubMed

Feild, Taylor S; Brodribb, Tim

2001-05-01

The effect of freezing on stem xylem hydraulic conductivity and leaf chlorophyll a fluorescence was measured in 12 tree and shrub species from a treeline heath in Tasmania, Australia. Reduction in stem hydraulic conductivity after a single freeze-thaw cycle was minimal in conifers and the vessel-less angiosperm species Tasmannia lanceolata (Winteraceae), whereas mean loss of conductivity in vessel-forming angiosperms fell in the range 17-83%. A positive linear relationship was observed between percentage loss of hydraulic conductivity by freeze-thaw and the average conduit diameter across all 12 species. This supports the hypothesis that large-diameter vascular conduits have a greater likelihood of freeze-thaw cavitation because larger bubbles are produced, which are more likely to expand under tension. Leaf frost tolerances, as measured by a 50% loss of maximum PSII quantum yield, varied from -6 to -13°C, indicating that these species were more frost-sensitive than plants from northern hemisphere temperate forest and treeline communities. There was no evidence of a relationship between frost tolerance of leaves and the resilience of stem water transport to freezing, suggesting that low temperature survival and the resistance of stem water transport to freezing are independently evolving traits. The results of this study bear on the ecological importance of stem freezing in the southern hemisphere treeline zones.

Slow-freezing-induced changes of birefringent structures in human oocytes are related to responsiveness to ovulation induction.

PubMed

Molinari, Emanuela; Revelli, Alberto; Racca, Cinzia; Delle Piane, Luisa; Massobrio, Marco

2010-05-01

The slow-freezing method is widely used to freeze human oocytes, both for fertility preservation and in routine IVF programmes. Slow freezing damages some of the cell's structures, including the meiotic spindle (MS) and the zona pellucida (ZP). Polarized light microscopy was used to study the variations induced by slow freezing on the MS and the ZP of human oocytes and to analyse the relationship between slow-freezing effects on the gamete and some clinical characteristics, such as age, body mass index and ovarian responsiveness to ovulation induction (expressed as total follicle-stimulating hormone dose/retrieved oocyte). Both the MS and the ZP (particularly its inner layer) underwent significant changes during slow-freezing procedure. The MS became thinner and structurally less organized (lower retardance) (P<0.001 and P<0.05, respectively), whereas the ZP became thicker and its inner layer lost structural organization (both P<0.05). These morphological changes were unrelated to the patient's age or body mass index, but ZP variations in thickness and retardance were significantly related to ovarian responsiveness (P=0.033 and P=0.026, respectively), suggesting that patients with a higher response to gonadotrophins produce oocytes better able to preserve their characteristics after freezing-thawing. Copyright (c) 2010 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Genes Upregulated in Winter Wheat (Triticum aestivum L.) during Mild Freezing and Subsequent Thawing Suggest Sequential Activation of Multiple Response Mechanisms.

PubMed

Skinner, Daniel Z

2015-01-01

Exposing fully cold-acclimated wheat plants to a mild freeze-thaw cycle of -3 °C for 24h followed by +3 °C for 24 or 48 h results in dramatically improved tolerance of subsequent exposure to sub-freezing temperatures. Gene enrichment analysis of crown tissue from plants collected before or after the -3 °C freeze or after thawing at +3 °C for 24 or 48 h revealed that many biological processes and molecular functions were activated during the freeze-thaw cycle in an increasing cascade of responses such that over 150 processes or functions were significantly enhanced by the end of the 48 h, post-freeze thaw. Nearly 2,000 individual genes were upregulated more than 2-fold over the 72 h course of freezing and thawing, but more than 70% of these genes were upregulated during only one of the time periods examined, suggesting a series of genes and gene functions were involved in activation of the processes that led to enhanced freezing tolerance. This series of functions appeared to include extensive cell signaling, activation of stress response mechanisms and the phenylpropanoid biosynthetic pathway, extensive modification of secondary metabolites, and physical restructuring of cell membranes. By identifying plant lines that are especially able to activate these multiple mechanisms it may be possible to develop lines with enhanced winterhardiness.

Effect of mixing time, freeze-drying and baking on phenolics, anthocyanins and antioxidant capacity of raspberry juice during processing of muffins.

PubMed

Rosales-Soto, Maria U; Powers, Joseph R; Alldredge, J Richard

2012-05-01

Consumption of baked products constitutes an important part of a daily breakfast considering that people are continually grabbing meals on the go. Among baked products, muffins rank third in breakfast products and attract a broad range of consumers. Incorporation of red raspberry juice into muffins can add value to the product while preserving health benefits to the consumer. The purpose of this study was to evaluate the effect of mixing time, freeze-drying and baking on the phenolic and anthocyanin contents and antioxidant capacity of raspberry juice during the preparation of muffins. Freeze-drying of raspberry batters reduced their phenolic content and antioxidant capacity regardless of mixing time. Non-freeze-dried raspberry batter mixed for 5 min had the highest phenolic content (0.88 mg gallic acid equivalent g(-1) dry matter (DM)). Non-freeze-dried raspberry muffins had the highest antioxidant capacity (0.041 µmol Trolox equivalent g(-1) DM). Freeze-dried raspberry batters mixed for 5 and 10 min had the highest anthocyanin content (0.065 mg cyanidin-3-glucoside g(-1) DM). Baking reduced the anthocyanin content of both non-freeze-dried and freeze-dried raspberry muffins. Despite the reduction in valuable compounds, muffin is a vehicle for the delivery of these compounds. Copyright © 2012 Society of Chemical Industry.

Development of freeze-dried miyeokguk, Korean seaweed soup, as space food sterilized by irradiation

NASA Astrophysics Data System (ADS)

Song, Beom-Seok; Park, Jin-Gyu; Kim, Jae-Hun; Choi, Jong-Il; Ahn, Dong-Hyun; Hao, Chen; Lee, Ju-Woon

2012-08-01

The purpose of this study was to evaluate microbial populations, Hunter's color values (L*, a*, b*) and the sensory quality of freeze-dried miyeokguk, Korean seaweed soup, in order to use it as space food. Microorganisms were not detected in non-irradiated freeze-dried miyeokguk within the detection limit of 1.00 log CFU/g. However, the microbial population in rehydrated miyeokguk was 7.01 log CFU/g after incubation at 35 °C for 48 h, indicating that freeze-dried miyeokguk was not sterilized by heat treatment during the preparation process. Bacteria in the freeze-dried miyeokguk were tentatively identified as Bacillus cereus, B. subtilis, Enterobacter hormaechei, and Ancinetobacter genomosp. using the 16S rDNA sequencing. In samples that were gamma-irradiated above 10 kGy, it was confirmed that all microorganisms were inactivated. Hunter's color values of the samples irradiated at doses less than 10 kGy were not significantly altered from their baseline appearance (p>0.05). Sensory evaluation showed that preference scores in all sensory properties decreased when freeze-dried miyeokguk was irradiated at doses greater than 10 kGy. Therefore, the results of this study suggest that gamma irradiation at 10 kGy is sufficient to sterilize freeze-dried miyeokguk without significant deterioration in the sensory quality, and thus, the freeze-dried and irradiated miyeokguk at 10 kGy fulfills the microbiological requirements as space food.

Effect of freezing on the rheological, chemical and colour properties of Serpa cheese.

PubMed

Alvarenga, Nuno; Canada, João; Sousa, Isabel

2011-02-01

The effect of freezing on the properties of a raw ewes'-milk semi-soft cheese (Serpa cheese) was studied using small amplitude oscillatory (SAOS) and texture measurements, colour and chemical parameters. The freezing was introduced at three different stages of the ripening process (28, 35 and 42 days), and the cheeses were maintained frozen for 12 months. Cheeses were submitted to a slow or fast freezing method, and to different storage temperatures: -10 and -20°C (three replicates for each set conditions). Chemical data showed that only the proteolysis indicators exhibited differences between frozen and non-frozen samples; frozen samples showed higher values of NPN than the non-frozen samples, indicating that the freezing process did not prevent the secondary proteolysis of cheese. Frozen samples showed a significantly (P<0·05) stronger structure than the non-frozen, as indicated by hardness. However, the differences between the frozen and non-frozen samples were not significantly for storage modulus (G' 1Hz) and loss tangent (tan δ 1Hz) (P>0·05). Freezing affected mainly colour parameters: frozen samples were more luminous, and more yellow-green. The results allowed us to conclude that the damages caused by freezing to cheese properties could be minimized if this type of storage is introduced at the end of ripening (42 d) using a freezing temperature of -20°C.

Dynamics and characteristics of soil temperature and moisture of active layer in central Tibetan Plateau

NASA Astrophysics Data System (ADS)

Zhao, L.; Hu, G.; Wu, X.; Tian, L.

2017-12-01

Research on the hydrothermal properties of active layer during the thawing and freezing processes was considered as a key question to revealing the heat and moisture exchanges between permafrost and atmosphere. The characteristics of freezing and thawing processes at Tanggula (TGL) site in permafrost regions on the Tibetan Plateau, the results revealed that the depth of daily soil temperature transmission was about 40 cm shallower during thawing period than that during the freezing period. Soil warming process at the depth above 140 cm was slower than the cooling process, whereas they were close below 140 cm depth. Moreover, the hydro-thermal properties differed significantly among different stages. Precipitation caused an obviously increase in soil moisture at 0-20 cm depth. The vertical distribution of soil moisture could be divided into two main zones: less than 12% in the freeze state and greater than 12% in the thaw state. In addition, coupling of moisture and heat during the freezing and thawing processes also showed that soil temperature decreased faster than soil moisture during the freezing process. At the freezing stage, soil moisture exhibited an exponential relationship with the absolute soil temperature. Energy consumed for water-ice conversion during the freezing process was 149.83 MJ/m2 and 141.22 MJ/m2 in 2011 and 2012, respectively, which was estimated by the soil moisture variation.

Freezing/Thawing without Cryoprotectant Damages Native but not Decellularized Porcine Renal Tissue

PubMed Central

Poornejad, Nafiseh; Frost, Timothy S; Scott, Daniel R; Elton, Brinden B; Reynolds, Paul R; Roeder, Beverly L; Cook, Alonzo D

2015-01-01

abstract Whole organ decellularization of porcine renal tissue and recellularization with a patient's own cells would potentially overcome immunorejection, which is one of the most significant problems with allogeneic kidney transplantation. However, there are obstacles to achieving this goal, including preservation of the decellularized extracellular matrix (ECM), identifying the proper cell types, and repopulating the ECM before transplantation. Freezing biological tissue is the best option to avoid spoilage; however, it may damage the structure of the tissue or disrupt cellular membranes through ice crystal formation. Cryoprotectants have been used to repress ice formation during freezing, although cell toxicity can still occur. The effect of freezing/thawing on native (n = 10) and decellularized (n = 10) whole porcine kidneys was studied without using cryoprotectants. Results showed that the elastic modulus of native kidneys was reduced by a factor of 22 (P < 0.0001) by freezing/thawing or decellularization, while the elastic modulus for decellularized ECM was essentially unchanged by the freezing/thawing process (p = 0.0636). Arterial pressure, representative of structural integrity, was also reduced by a factor of 52 (P < 0.0001) after freezing/thawing for native kidneys, compared to a factor of 43 (P < 0.0001) for decellularization and a factor of 4 (P < 0.0001) for freezing/thawing decellularized structures. Both freezing/thawing and decellularization reduced stiffness, but the reductions were not additive. Investigation of the microstructure of frozen/thawed native and decellularized renal tissues showed increased porosity due to cell removal and ice crystal formation. Orcein and Sirius staining showed partial damage to elastic and collagen fibers after freezing/thawing. It was concluded that cellular damage and removal was more responsible for reducing stiffness than fibril destruction. Cell viability and growth were demonstrated on decellularized frozen/thawed and non-frozen samples using human renal cortical tubular epithelial (RCTE) cells over 12 d. No adverse effect on the ability to recellularize after freezing/thawing was observed. It is recommended that porcine kidneys be frozen prior to decellularization to prevent contamination, and after decellularization to prevent protein denaturation. Cryoprotectants may still be necessary, however, during storage and transportation after recellularization. PMID:25730294

Isolation of a Soluble Component of Plasmodium berghei Which Induces Immunity in Rats

PubMed Central

Grothaus, G. D.; Kreier, J. P.

1980-01-01

Soluble material was obtained from sonically freed plasmodiae by three procedures. Two procedures, cryo-impacting and freeze-thawing, were evaluated for their ability to disrupt the parasites and release soluble material. The soluble materials obtained by these procedures were compared to materials washed from the surfaces of sonically freed parasites. Between 35 and 40% of the total parasite protein was solubilized by freeze-thawing or cryo-impacting. One cycle of freeze-thawing released nearly as much protein as could be released by this method, and additional cycles of freeze-thawing had little additional effect. Cryo-impacting solubilized only a small amount of protein in addition to that which was released by the cycle of freeze-thawing inherent in the procedure. Reductions in the packed cell volume of the material remaining after freeze-thawing or cryo-impacting indicate that the insoluble fragments are broken into smaller pieces as treatment is extended. Electron microscopy of 30-s cryo-impacted and three-times freeze-thawed parasites revealed membrane fragments similar in appearance. Patterns obtained by polyacrylamide gel electrophoresis of the soluble material from freeze-thawed and cryo-impacted parasites were also similar, and approximately 13 protein bands were demonstrated. The material washed from the surfaces of the free parasites, on the other hand, resolved into only two to four major bands on the gel columns. In immunization studies, the soluble and insoluble fractions obtained by freeze-thawing or cryo-impacting and the material washed from the surfaces of the parasites all stimulated a protective immune response. On the basis of the amount of protein required to stimulate roughly comparable immunity, the soluble fraction obtained by freeze-thawing or cryo-impacting free parasites was about twice as potent an immunogen as was the insoluble fraction. The material obtained by gentle washing of the freed parasites was approximately 20 times as potent an immunogen as were the freed parasites and about 7 times as potent as the soluble material obtained by freeze-thawing or cryo-impacting. Images Fig. 1 Fig. 2 PMID:6991439

Model simulations with COSMO-SPECS: impact of heterogeneous freezing modes and ice nucleating particle types on ice formation and precipitation in a deep convective cloud

NASA Astrophysics Data System (ADS)

Diehl, Karoline; Grützun, Verena

2018-03-01

In deep convective clouds, heavy rain is often formed involving the ice phase. Simulations were performed using the 3-D cloud resolving model COSMO-SPECS with detailed spectral microphysics including parameterizations of homogeneous and three heterogeneous freezing modes. The initial conditions were selected to result in a deep convective cloud reaching 14 km of altitude with strong updrafts up to 40 m s-1. At such altitudes with corresponding temperatures below -40 °C the major fraction of liquid drops freezes homogeneously. The goal of the present model simulations was to investigate how additional heterogeneous freezing will affect ice formation and precipitation although its contribution to total ice formation may be rather low. In such a situation small perturbations that do not show significant effects at first sight may trigger cloud microphysical responses. Effects of the following small perturbations were studied: (1) additional ice formation via immersion, contact, and deposition modes in comparison to solely homogeneous freezing, (2) contact and deposition freezing in comparison to immersion freezing, and (3) small fractions of biological ice nucleating particles (INPs) in comparison to higher fractions of mineral dust INP. The results indicate that the modification of precipitation proceeds via the formation of larger ice particles, which may be supported by direct freezing of larger drops, the growth of pristine ice particles by riming, and by nucleation of larger drops by collisions with pristine ice particles. In comparison to the reference case with homogeneous freezing only, such small perturbations due to additional heterogeneous freezing rather affect the total precipitation amount. It is more likely that the temporal development and the local distribution of precipitation are affected by such perturbations. This results in a gradual increase in precipitation at early cloud stages instead of a strong increase at later cloud stages coupled with approximately 50 % more precipitation in the cloud center. The modifications depend on the active freezing modes, the fractions of active INP, and the composition of the internal mixtures in the drops.

Effects of whole body (56)Fe radiation on contextual freezing and Arc-positive cells in the dentate gyrus.

PubMed

Raber, Jacob; Allen, Antiño R; Rosi, Susanna; Sharma, Sourabh; Dayger, Catherine; Davis, Matthew J; Fike, John R

2013-06-01

The space radiation environment contains high-energy charged particles such as (56)Fe, which could pose a significant hazard to hippocampal function in astronauts during and after the mission(s). The mechanisms underlying impairments in cognition are not clear but might involve alterations in the percentage of neurons in the dentate gyrus expressing the plasticity-related immediate early gene Arc. Previously, we showed effects of cranial (56)Fe irradiation on hippocampus-dependent contextual freezing and on the percentage of Arc-positive cells in the enclosed, but not free, blade. Because it is unclear whether whole body (56)Fe irradiation causes similar effects on these markers of hippocampal function, in the present study we quantified the effects of whole body (56)Fe irradiation (600MeV, 0.5 or 1Gy) on hippocampus-dependent and hippocampus-independent cognitive performance and determined whether these effects were associated with changes in Arc expression in the enclosed and free blades of the dentate gyrus. Whole body (56)Fe irradiation impacted contextual but not cued fear freezing and the percentage of Arc-positive cells in the enclosed and free blades. In mice tested for contextual freezing, there was a correlation between Arc-positive cells in the enclosed and free blades. In addition, in mice irradiated with 0.5Gy, contextual freezing in the absence of aversive stimuli correlated with the percentage of Arc-positive cells in the enclosed blade. In mice tested for cued freezing, there was no correlation between Arc-positive cells in the enclosed and free blades. In contrast, cued freezing in the presence or absence of aversive stimuli correlated with Arc-positive cells in the free blade. In addition, in mice irradiated with 1Gy cued freezing in the absence of aversive stimuli correlated with the percentage of Arc-positive neurons in the free blade. These data indicate that while whole body (56)Fe radiation affects contextual freezing and Arc-positive cells in the dentate gyrus, the enclosed blade might be more important for contextual freezing while the free blade might be more important for cued freezing. Copyright © 2013 Elsevier B.V. All rights reserved.

Freeze-thaw performance testing of whole concrete railroad ties.

DOT National Transportation Integrated Search

2013-10-01

Freezing and thawing durability tests of prestressed concrete ties are normally performed according to ASTM C666 specifications. Small specimens are cut from the shoulders of concrete ties and tested through 300 cycles of freezing and thawing. Saw-cu...

Effect of the Winter Wheat Cheyenne 5A Substituted Chromosome on Dynamics of Abscisic Acid and Cytokinins in Freezing-Sensitive Chinese Spring Genetic Background

PubMed Central

Kalapos, Balázs; Novák, Aliz; Dobrev, Petre; Vítámvás, Pavel; Marincs, Ferenc; Galiba, Gábor; Vanková, Radomira

2017-01-01

The effect of short- and long-term cold treatment on the abscisic acid (ABA) and cytokinin (CK) metabolism, and their main biosynthesis- and signaling-related genes were investigated in freezing-sensitive and freezing-tolerant wheat genotypes. Varieties Cheyenne and Chinese Spring substituted with the 5A Cheyenne chromosome, which represented freezing-tolerant genotypes, were compared with the freezing-sensitive Chinese Spring. Hormone levels and gene expression data indicated that the short- and long-term cold treatments are associated with specific regulation of the accumulation of cold-protective proteins and phytohormone levels, as well as the expression profiles of the hormone-related genes. The significant differences were observed between the genotypes, and between their leaf and crown tissues, too. The level of dehydrins, including WCS120 protein, and expression of WCS120 gene were considerably higher in the freezing-tolerant genotypes after 21 days of cold treatment. Expression of Cor14b and CBF14, cold-responsive regulator genes, was increased by cold treatment in all genotypes, to higher extent in freezing-tolerant genotypes. Cluster analysis revealed that the tolerant genotypes had a similar response to cold treatment, regarding expression of the ABA and CK metabolic genes, as well as hormone levels in leaves. As far as hormone levels in crowns are concerned, however, the strongly freezing-tolerant Cheyenne variety clustered separately from the Chinese Spring and the substitution line, which were more similar to each other after both 1 and 21 days of cold treatment than to Cheyenne. Based on these results we concluded that the 5A chromosome of wheat might have both a direct and an indirect impact on the phytohormone-dependent cold-induced freezing tolerance. Based on the gene expression data, novel genetic markers could be developed, which may be used to determine the freezing tolerance level in a wide range of wheat varieties. PMID:29238355

Effect of controlled ice nucleation on primary drying stage and protein recovery in vials cooled in a modified freeze-dryer.

PubMed

Passot, Stéphanie; Tréléa, Ioan Cristian; Marin, Michèle; Galan, Miquel; Morris, G John; Fonseca, Fernanda

2009-07-01

The freezing step influences lyophilization efficiency and protein stability. The main objective of this work was to investigate the impact on the primary drying stage of an ultrasound controlled ice nucleation technology, compared with usual freezing protocols. Lyophilization cycles involving different freezing protocols (applying a constant shelf cooling rate of 1 degrees C/min or 0.2 degrees C/min, putting vials on a precooled shelf, and controlling nucleation by ultrasounds or by addition of a nucleating agent) were performed in a prototype freeze-dryer. Three protective media including sucrose or maltodextrin and differing by their thermal properties and their ability to preserve a model protein (catalase) were used. The visual aspect of the lyophilized cake, residual water content, and enzymatic activity recovery of catalase were assessed after each lyophilization cycle and after 1 month of storage of the lyophilized product at 4 degrees C and 25 degrees C. The freezing protocols allowing increasing nucleation temperature (precooled shelf and controlled nucleation by using ultrasounds or a nucleating agent) induced a faster sublimation step and higher sublimation rate homogeneity. Whatever the composition of the protective medium, applying the ultrasound technology made it possible to decrease the sublimation time by 14%, compared with the freezing method involving a constant shelf cooling rate of 1 degrees C/min. Concerning the enzyme activity recovery, the impact of the freezing protocol was observed only for the protective medium involving maltodextrin, a less effective protective agent than sucrose. Higher activity recovery results were obtained after storage when the ultrasound technology or the precooled shelf method was applied. Controlling ice nucleation during the freezing step of the lyophilization process improved the homogeneity of the sublimation rates, which will, in turn, reduce the intervial heterogeneity. The freeze-dryer prototype including the system of controlled nucleation by ultrasounds appears to be a promising tool in accelerating sublimation and improving intrabatch homogeneity.

Freeze-Thaw Cycles and Soil Biogeochemistry: Implications for Greenhouse Gas emission

NASA Astrophysics Data System (ADS)

Rezanezhad, F.; Milojevic, T.; Oh, D. H.; Parsons, C. T.; Smeaton, C. M.; Van Cappellen, P.

2016-12-01

Freeze-thaw cycles represent a major natural climate forcing acting on soils at middle and high latitudes. Repeated freezing and thawing of soils changes their physical properties, geochemistry, and microbial community structure, which together govern the biogeochemical cycling of carbon and nutrients. In this presentation, we focus on how freeze-thaw cycles regulate carbon and nitrogen cycling and how these transformations influence greenhouse gas (GHG) fluxes. We present a novel approach, which combines the acquisition of physical and chemical data in a newly developed experimental soil column system. This system simulates realistic soil temperature profiles during freeze-thaw cycles. A high-resolution, Multi-Fiber Optode (MuFO) microsensor technique was used to detect oxygen (O2) continuously in the column at multiple depths. Surface and subsurface changes to gas and aqueous phase chemistry were measured to delineate the pathways and quantify soil respiration rates during freeze-thaw cycles. The results indicate that the time-dependent release of GHG from the soil surface is influenced by a combination of two key factors. Firstly, fluctuations in temperature and O2 availability affect soil biogeochemical activity and GHG production. Secondly, the recurrent development of a physical ice barrier prevents exchange of gaseous compounds between the soil and atmosphere during freezing conditions; removal of this barrier during thaw conditions increases GHG fluxes. During freezing, O2 levels in the unsaturated zone decreased due to restricted gas exchange with the atmosphere. As the soil thawed, O2 penetrated deeper into the soil enhancing the aerobic mineralization of organic carbon and nitrogen. Additionally, with the onset of thawing a pulse of gas flux occurred, which is attributed to the build-up of respiratory gases in the pore space during freezing. The latter implies enhanced anaerobic respiration as O2 supply ceases when the upper soil layer freezes.

Remote sensing of freeze-thaw transitions in Arctic soils using the complex resistivity method

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wu, Yuxin; Hubbard, Susan S; Ulrich, Craig

2013-01-01

Our ability to monitor freeze - thaw transitions is critical to developing a predictive understanding of biogeochemical transitions and carbon dynamics in high latitude environments. In this study, we conducted laboratory column experiments to explore the potential of the complex resistivity method for monitoring the freeze - thaw transitions of the arctic permafrost soils. Samples for the experiment were collected from the upper active layer of Gelisol soils at the Barrow Environmental Observatory, Barrow Alaska. Freeze - thaw transitions were induced through exposing the soil column to controlled temperature environments at 4 C and -20 C. Complex resistivity and temperaturemore » measurements were collected regularly during the freeze - thaw transitions using electrodes and temperature sensors installed along the column. During the experiments, over two orders of magnitude of resistivity variations were observed when the temperature was increased or decreased between -20 C and 0 C. Smaller resistivity variations were also observed during the isothermal thawing or freezing processes that occurred near 0 C. Single frequency electrical phase response and imaginary conductivity at 1 Hz were found to be exclusively related to the unfrozen water in the soil matrix, suggesting that these geophysical 24 attributes can be used as a proxy for the monitoring of the onset and progression of the freeze - thaw transitions. Spectral electrical responses and fitted Cole Cole parameters contained additional information about the freeze - thaw transition affected by the soil grain size distribution. Specifically, a shift of the observed spectral response to lower frequency was observed during isothermal thawing process, which we interpret to be due to sequential thawing, first from fine then to coarse particles within the soil matrix. Our study demonstrates the potential of the complex resistivity method for remote monitoring of freeze - thaw transitions in arctic soils. Although conducted at the laboratory scale, this study provides the foundation for exploring the potential of the complex resistivity signals for monitoring spatiotemporal variations of freeze - thaw transitions over field-relevant scales.« less

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

PubMed Central

Bobik, Krzysztof; Dunlap, John R.; Burch-Smith, Tessa M.

2014-01-01

Since the 1940s transmission electron microscopy (TEM) has been providing biologists with ultra-high resolution images of biological materials. Yet, because of laborious and time-consuming protocols that also demand experience in preparation of artifact-free samples, TEM is not considered a user-friendly technique. Traditional sample preparation for TEM used chemical fixatives to preserve cellular structures. High-pressure freezing is the cryofixation of biological samples under high pressures to produce very fast cooling rates, thereby restricting ice formation, which is detrimental to the integrity of cellular ultrastructure. High-pressure freezing and freeze substitution are currently the methods of choice for producing the highest quality morphology in resin sections for TEM. These methods minimize the artifacts normally associated with conventional processing for TEM of thin sections. After cryofixation the frozen water in the sample is replaced with liquid organic solvent at low temperatures, a process called freeze substitution. Freeze substitution is typically carried out over several days in dedicated, costly equipment. A recent innovation allows the process to be completed in three hours, instead of the usual two days. This is typically followed by several more days of sample preparation that includes infiltration and embedding in epoxy resins before sectioning. Here we present a protocol combining high-pressure freezing and quick freeze substitution that enables plant sample fixation to be accomplished within hours. The protocol can readily be adapted for working with other tissues or organisms. Plant tissues are of special concern because of the presence of aerated spaces and water-filled vacuoles that impede ice-free freezing of water. In addition, the process of chemical fixation is especially long in plants due to cell walls impeding the penetration of the chemicals to deep within the tissues. Plant tissues are therefore particularly challenging, but this protocol is reliable and produces samples of the highest quality. PMID:25350384

Roles of testosterone and amygdaloid LTP induction in determining sex differences in fear memory magnitude.

PubMed

Chen, Li-Shen; Tzeng, Wen-Yu; Chuang, Jia-Ying; Cherng, Chianfang G; Gean, Po-Wu; Yu, Lung

2014-08-01

Women are thought to form fear memory more robust than men do and testosterone is suspected to play a role in determining such a sex difference. Mouse cued fear freezing was used to study the sex-related susceptibility and the role of testosterone in fear memory in humans. A 75-dB tone was found to provoke weak freezing, while 0.15-mA and 0.20-mA footshock caused strong freezing responses. No sex differences were noticed in the tone- or footshock-induced (naïve fear) freezing. Following the conditionings, female mice exhibited greater tone (cued fear)-induced freezing than did male mice. Nonetheless, female mice demonstrated indistinctive cued fear freezing across the estrous phases and ovariectomy did not affect such freezing in female mice. Orchidectomy enhanced the cued fear freezing in male mice. Systemic testosterone administrations and an intra-lateral nucleus of amygdala (LA) testosterone infusion diminished the cued fear freezing in orchidectomized male mice, while pretreatment with flutamide (Flu) eradicated these effects. Long-term potentiation (LTP) magnitude in LA has been known to correlate with the strength of the cued fear conditioning. We found that LA LTP magnitude was indeed greater in female than male mice. Orchidectomy enhanced LTP magnitude in males' LA, while ovariectomy decreased LTP magnitude in females' LA. Testosterone decreased LTP magnitude in orchidectomized males' LA and estradiol enhanced LTP magnitude in ovariectomized females' LA. Finally, male mice had lower LA GluR1 expression than female mice and orchidectomy enhanced the GluR1 expression in male mice. These findings, taken together, suggest that testosterone plays a critical role in rendering the sex differences in the cued fear freezing and LA LTP. Testosterone is negatively associated with LA LTP and the cued fear memory in male mice. However, ovarian hormones and LA LTP are loosely associated with the cued fear memory in female mice. Copyright © 2014 Elsevier Inc. All rights reserved.

Reducing the loss of vaccines from accidental freezing in the cold chain: the experience of continuous temperature monitoring in Tunisia.

PubMed

Lloyd, John; Lydon, Patrick; Ouhichi, Ramzi; Zaffran, Michel

2015-02-11

Accidental freezing of vaccines is a growing threat and a real risk for national immunization programs when the potency of many vaccines can be compromised if these are exposed to sub-zero temperatures in the cold chain. In Tunisia, this issue is compounded by using sub-standard domestic cold chain equipment instead of equipping the program with medical refrigerators designed specifically for storing vaccines and temperature sensitive pharmaceuticals. Against this backdrop, this paper presents the findings of a demonstration project conducted in Tunisia in 2012 that tested the impact of introducing several freeze prevention solutions to mitigate the risk of accidental freezing of vaccines. The main finding is that, despite the continued use of underperforming domestic refrigerators, continuous temperature monitoring using new technologies combined with other technological interventions significantly reduced the prevalence of accidental exposure to freezing temperatures. These improvements were noticed for cold chain storage at regional, district and health center levels, and during the transport legs that were part of the demonstration conducted in the regions of Kasserine in the South-Eastern part of Tunisia. Subsequent to introducing these freeze prevention solutions, the incidence of freeze alarms was reduced and the percent of time the temperatures dropped below the 2 °C recommended threshold. The incidence of freeze alarms at health center level was reduced by 40%. Lastly, the solutions implemented reduced risk of freezing during transport from 13.8% to 1.7%. Although the solution implemented is not optimal in the longer term because domestic refrigerators are used extensively in district stores and health centers, the risk of accidental freezing is significantly reduced by introducing the practice of continuous temperature monitoring as a standard. The management of the cold chain equipment was strengthened as a result which helps protect the potency of vaccines to the areas of most difficult access. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Optimization of the freezing process for hematopoietic progenitor cells: effect of precooling, initial dimethyl sulfoxide concentration, freezing program, and storage in vapor-phase or liquid nitrogen on in vitro white blood cell quality.

PubMed

Dijkstra-Tiekstra, Margriet J; Setroikromo, Airies C; Kraan, Marcha; Gkoumassi, Effimia; de Wildt-Eggen, Janny

2014-12-01

Adding dimethyl sulfoxide (DMSO) to hematopoietic progenitor cells (HPCs) causes an exothermic reaction, potentially affecting their viability. The freezing method might also influence this. The aim was to investigate the effect of 1) precooling of DMSO and plasma (D/P) and white blood cell (WBC)-enriched product, 2) DMSO concentration of D/P, 3) freezing program, and 4) storage method on WBC quality. WBC-enriched product without CD34+ cells was used instead of HPCs. This was divided into six or eight portions. D/P (20 or 50%; precooled or room temperature [RT]) was added to the WBC-enriched product (precooled or RT), resulting in 10% DMSO, while monitoring temperature. The product was frozen using controlled-rate freezing ("fast-rate" or "slow-rate") and placed in vapor-phase or liquid nitrogen. After thawing, WBC recovery and viability were determined. Temperature increased most for precooled D/P to precooled WBC-enriched product, without influence of 20 or 50% D/P, but remained for all variations below 30°C. WBC recovery for both freezing programs was more than 95%. Recovery of WBC viability was higher for slow-rate freezing compared to fast-rate freezing (74% vs. 61%; p < 0.05) and also for 50% compared to 20% D/P (two test variations). Effect of precooling D/P or WBC-enriched product and of storage in vapor-phase or liquid nitrogen was marginal. Based on these results, precooling is not necessary. Fifty percent D/P is preferred over 20% D/P. Slow-rate freezing is preferred over fast-rate freezing. For safety reasons storage in vapor-phase nitrogen is preferred over storage in liquid nitrogen. Additional testing using real HPCs might be necessary. © 2014 AABB.

Saccharomyces cerevisiae glycerol/H+ symporter Stl1p is essential for cold/near-freeze and freeze stress adaptation. A simple recipe with high biotechnological potential is given

PubMed Central

2010-01-01

Background Freezing is an increasingly important means of preservation and storage of microbial strains used for many types of industrial applications including food processing. However, the yeast mechanisms of tolerance and sensitivity to freeze or near-freeze stress are still poorly understood. More knowledge on this regard would improve their biotechnological potential. Glycerol, in particular intracellular glycerol, has been assigned as a cryoprotectant, also important for cold/near-freeze stress adaptation. The S. cerevisiae glycerol active transporter Stl1p plays an important role on the fast accumulation of glycerol. This gene is expressed under gluconeogenic conditions, under osmotic shock and stress, as well as under high temperatures. Results We found that cells grown on STL1 induction medium (YPGE) and subjected to cold/near-freeze stress, displayed an extremely high expression of this gene, also visible at glycerol/H+ symporter activity level. Under the same conditions, the strains harbouring this transporter accumulated more than 400 mM glycerol, whereas the glycerol/H+ symporter mutant presented less than 1 mM. Consistently, the strains able to accumulate glycerol survive 25-50% more than the stl1Δ mutant. Conclusions In this work, we report the contribution of the glycerol/H+ symporter Stl1p for the accumulation and maintenance of glycerol intracellular levels, and consequently cell survival at cold/near-freeze and freeze temperatures. These findings have a high biotechnological impact, as they show that any S. cerevisiae strain already in use can become more resistant to cold/freeze-thaw stress just by simply adding glycerol to the broth. The combination of low temperatures with extracellular glycerol will induce the transporter Stl1p. This solution avoids the use of transgenic strains, in particular in food industry. PMID:21047428

Effect of freezing temperature on the color of frozen salmon.

PubMed

Ottestad, Silje; Enersen, Grethe; Wold, Jens Petter

2011-09-01

New freezing methods developed with the purpose of improved product quality after thawing can sometimes be difficult to get accepted in the market. The reason for this is the formation of ice crystals that can give the product a temporary color loss and make it less appealing. We have here used microscopy to study ice crystal size as a function of freezing temperature by investigating the voids in the cell tissue left by the ice crystals. We have also investigated how freezing temperature affects the color and the visible absorption spectra of frozen salmon. Freezing temperatures previously determined to be the best for quality after thawing (-40 to -60 °C) were found to cause a substantial loss in perceived color intensity during frozen state. This illustrated the conflict between optimal freezing temperatures with respect to quality after thawing against visual appearance during frozen state. Low freezing temperatures gave many small ice crystals, increased light scattering and an increased absorption level for all wavelengths in the visible region. Increased astaxanthin concentration on the other hand would give higher absorption at 490 nm. The results showed a clear potential of using visible interactance spectroscopy to differentiate between poor product coloration due to lack of pigmentation and temporary color loss due to light scattering by ice crystal. This type of measurements could be a useful tool in the development of new freezing methods and to monitor ice crystal growth during frozen storage. It could also potentially be used by the industry to prove good product quality. In this article we have shown that freezing food products at intermediate to low temperatures (-40 to -80 °C) can result in paler color during frozen state, which could affect consumer acceptance. We have also presented a spectroscopic method that can separate between poor product color and temporary color loss due to freezing. © 2011 Institute of Food Technologists®

Proteomic analysis of spring freeze-stress responsive proteins in leaves of bread wheat (Triticum aestivum L.).

PubMed

Han, Qiaoxia; Kang, Guozhang; Guo, Tiancai

2013-02-01

Following three-day exposure to -5 °C simulated spring freeze stress, wheat plants at the anther connective tissue formation phase of spike development displayed the drooping and wilting of leaves and markedly increased rates of relative electrolyte leakage. We analysed freeze-stress responsive proteins in wheat leaves at one and three days following freeze-stress exposure, using two-dimensional electrophoresis and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. Our results indicate that out of 75 protein spots successfully identified under freeze-stress conditions 52 spots were upregulated and 18 were downregulated. These spring freeze-stress responsive proteins were involved in signal transduction, stress/defence/detoxification, protein metabolism (i.e. translation, processing, and degradation), photosynthesis, amino acid metabolism, carbohydrate metabolism, and energy pathways, and may therefore be functionally relevant for many biological processes. The enhanced accumulation of signal transduction proteins such as a C2H2 zinc finger protein, stress/defence/detoxification proteins including LEA-related COR protein, disease resistance protein, Cu/Zn superoxide dismutase, and two ascorbate peroxidases may play crucial roles in the mechanisms of response to spring freeze stress in wheat plants. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

Reproductive technology and the life course: current debates and research in social egg freezing.

PubMed

Baldwin, Kylie; Culley, Lorraine; Hudson, Nicky; Mitchell, Helene

2014-09-01

While there are currently few confirmed births from previously frozen eggs in the UK, the improved outcomes of new technologies of vitrification and intra-cytoplasmic sperm injection (ICSI) have led to the marketing of egg freezing for non-medical reasons, whereby women are offered the possibility of preserving their eggs until such time as they wish to have a child. Non-academic commentary on this phenomenon has commonly constructed it within a neo-liberal discourse of 'choice', whereby women are positioned as choosing to 'delay motherhood,' often for reasons relating to their careers. However, there have been relatively few research studies which explore either women's awareness and understandings of social egg freezing or the reasons why women consider or undertake egg freezing. This paper summarises the current academic discussion surrounding social egg freezing and outlines the limited body of empirical literature identified from a systematic search of relevant databases. The potential benefits and harms of social egg freezing and the ethical issues it raises are well rehearsed in existing literature, but there is limited empirical evidence about who is accessing social egg freezing, why they are taking this option and what their experiences and future intentions are.

Extracellular ice phase transitions in insects.

PubMed

Hawes, T C

2014-01-01

At temperatures below their temperature of crystallization (Tc), the extracellular body fluids of insects undergo a phase transition from liquid to solid. Insects that survive the transition to equilibrium (complete freezing of the body fluids) are designated as freeze tolerant. Although this phenomenon has been reported and described in many Insecta, current nomenclature and theory does not clearly delineate between the process of transition (freezing) and the final solid phase itself (the frozen state). Thus freeze tolerant insects are currently, by convention, described in terms of the temperature at which the crystallization of their body fluids is initiated, Tc. In fact, the correct descriptor for insects that tolerate freezing is the temperature of equilibrium freezing, Tef. The process of freezing is itself a separate physical event with unique physiological stresses that are associated with ice growth. Correspondingly there are a number of insects whose physiological cryo-limits are very specifically delineated by this transitional envelope. The distinction also has considerable significance for our understanding of insect cryobiology: firstly, because the ability to manage endogenous ice growth is a fundamental segregator of cryotype; and secondly, because our understanding of internal ice management is still largely nascent.

Cryopreservation of human sperm: efficacy and use of a new nitrogen-free controlled rate freezer versus liquid nitrogen vapour freezing.

PubMed

Creemers, E; Nijs, M; Vanheusden, E; Ombelet, W

2011-12-01

Preservation of spermatozoa is an important aspect of assisted reproductive medicine. The aim of this study was to investigate the efficacy and use of a recently developed liquid nitrogen and cryogen-free controlled rate freezer and this compared with the classical liquid nitrogen vapour freezing method for the cryopreservation of human spermatozoa. Ten patients entering the IVF programme donated semen samples for the study. Samples were analysed according to the World Health Organization guidelines. No significant difference in total sperm motility after freeze-thawing between the new technique and classical technique was demonstrated. The advantage of the new freezing technique is that it uses no liquid nitrogen during the freezing process, hence being safer to use and clean room compatible. Investment costs are higher for the apparatus but running costs are only 1% in comparison with classical liquid nitrogen freezing. In conclusion, post-thaw motility of samples frozen with the classical liquid nitrogen vapour technique was comparable with samples frozen with the new nitrogen-free freezing technique. This latter technique can thus be a very useful asset to the sperm cryopreservation laboratory. © 2011 Blackwell Verlag GmbH.

Freezing behavior of adherent neuron-like cells and morphological change and viability of post-thaw cells.

PubMed

Uemura, Makoto; Ishiguro, Hiroshi

2015-04-01

Freezing of nerve cells forming a neuronal network has largely been neglected, despite the fact that the cryopreservation of nerve cells benefits the study of cells in the areas of medicine and poison screening. Freezing of nerve cells is also attractive for studying cell morphology because of the characteristic long, thread-like neurites extending from the cell body. In the present study, freezing of neuron-like cells adhering to the substrate (differentiated PC12 cells), in physiological saline, was investigated in order to understand the fundamental freezing and thawing characteristics of nerve cells with neurites. The microscopic freezing behavior of cells under different cooling rates was observed. Next, the post-thaw morphological changes in the cells, including the cytoskeleton, were investigated and post-thaw cell viability was evaluated by dye exclusion using propidium iodide. Two categories of morphological changes, beading and shortening of the neurites, were found and quantified. Also, the morphological changes of neurites due to osmotic stress from sodium chloride were studied to gain a better understanding of causation. The results showed that morphological changes and cell death were promoted with a decrease in end temperature during freezing. Copyright © 2015 Elsevier Inc. All rights reserved.

Role of ice structuring proteins on freezing-thawing cycles of pasta sauces.

PubMed

Calderara, Marianna; Deorsola, Fabio A; Bensaid, Samir; Fino, Debora; Russo, Nunzio; Geobaldo, Francesco

2016-12-01

The freezing of the food is one of the most important technological developments for the storage of food in terms of quality and safety. The aim of this work was to study the role of an ice structuring protein (ISP) on freezing-thawing cycles of different solutions and commercial Italian pasta sauces. Ice structuring proteins were related to the modification of the structure of ice. The results showed that the freezing time of an aqueous solution containing the protein was reduced to about 20% with respect to a pure water solution. The same effect was demonstrated in sugar-containing solutions and in lipid-containing sauces. The study proved a specific role of ISP during thawing, inducing a time decrease similar to that of freezing and even more important in the case of tomato-based sauces. This work demonstrated the role of ISP in the freezing-thawing process, showing a significant reduction of processing in the freezing and thawing phase by adding the protein to pure water and different sugar-, salt- and lipid-containing solutions and commercial sauces, with considerable benefits for the food industry in terms of costs and food quality.

Identification of Arabidopsis mutants with altered freezing tolerance.

PubMed

Perea-Resa, Carlos; Salinas, Julio

2014-01-01

Low temperature is an important determinant in the configuration of natural plant communities and defines the range of distribution and growth of important crops. Some plants, including Arabidopsis, have evolved sophisticated adaptive mechanisms to tolerate low and freezing temperatures. Central to this adaptation is the process of cold acclimation. By means of this process, many plants from temperate regions are able to develop or increase their freezing tolerance in response to low, nonfreezing temperatures. The identification and characterization of factors involved in freezing tolerance are crucial to understand the molecular mechanisms underlying the cold acclimation response and have a potential interest to improve crop tolerance to freezing temperatures. Many genes implicated in cold acclimation have been identified in numerous plant species by using molecular approaches followed by reverse genetic analysis. Remarkably, however, direct genetic analyses have not been conveniently exploited in their capacity for identifying genes with pivotal roles in that adaptive response. In this chapter, we describe a protocol for evaluating the freezing tolerance of both non-acclimated and cold-acclimated Arabidopsis plants. This protocol allows the accurate and simple screening of mutant collections for the identification of novel factors involved in freezing tolerance and cold acclimation.

A New Freezing Method Using Pre-Dehydration by Microwave-Vacuum Drying

NASA Astrophysics Data System (ADS)

Tsuruta, Takaharu; Hamidi, Nurkholis

Partial dehydration by microwave-vacuum drying has been applied to tuna and strawberry in order to reduce cell-damages caused by the formation of large ice-crystals during freezing. The samples were subjected to microwave vacuum drying at pressure of 5 kPa and temperature less than 27°C to remove small amount of water prior to freezing. The tuna were cooled by using the freezing chamber at temperature -50°C or -150°C, while the strawberries were frozen at temperature -30°C or -80°C, respectively. The temperature transients in tuna showed that removing some water before freezing made the freezing time shorter. The observations of ice crystal clearly indicated that rapid cooling and pre-dehydration prior to freezing were effective in minimizing the size of ice crystal. It is also understood that the formation of large ice crystals has a close relation to the cell damages. After thawing, the observation of microstructure was done on the tuna and strawberry halves. The pre-dehydrated samples showed a better structure than the un-dehydrated one. It is concluded that the pre-dehydration by microwave-vacuum drying is one promising method for the cryo-preservation of foods.

Simulations of Polar Stratospheric Clouds and Denitrification Using Laboratory Freezing Rates

NASA Technical Reports Server (NTRS)

Drdla, Katja; Tabazadeh, Azadeh; Gore, Warren J. (Technical Monitor)

2001-01-01

During the 1999-2000 Arctic winter, the SAGE (Stratospheric Aerosol and Gas Experiment) III Ozone Loss and Validation Experiment (SOLVE) provided evidence of widespread solid-phase polar stratospheric clouds (PSCs) accompanied by severe nitrification. Previous simulations have shown that a freezing process occurring at temperatures above the ice frost point is necessary to explain these observations. In this work, the nitric acid freezing rates measured by Salcedo et al. and discussed by Tabazadeh et al. have been examined. These freezing rates have been tested in winter-long microphysical simulations of the 1999-2000 Arctic vortex evolution in order to determine whether they can explain the observations. A range of cases have been explored, including whether the PSC particles are composed of nitric acid dihydrate or trihydrate, whether the freezing process is a bulk process or occurs only on the particle surfaces, and uncertainties in the derived freezing rates. Finally, the possibility that meteoritic debris enhances the freezing rate has also been examined. The results of these simulations have been compared with key PSC and denitrification measurements made by the SOLVE campaign. The cases that best reproduce the measurements will he highlighted, with a discussion of the implications for our understanding of PSCs.

Beneficial effect of directional freezing on in vitro viability of cryopreserved sheep whole ovaries and ovarian cortical slices.

PubMed

Maffei, S; Pennarossa, G; Brevini, T A L; Arav, A; Gandolfi, F

2014-01-01

Does directional freezing improve the structural and functional integrity of ovarian fragments compared with conventional slow freezing and to whole ovary cryopreservation? Compared with slow freezing, the use of directional freezing significantly improves all structural and functional parameters of ovarian fragments assessed in vitro and, overall, whole ovaries were better preserved than ovarian fragments. Directional freezing has been developed to provide an alternative way to cryopreserve large biological samples and it is known to improve the structural and functional integrity of whole ovaries. Conventional slow freezing of ovarian fragments is the procedure more widely used in clinical settings but it causes substantial structural damage that limits the functional period after transfer back into the patient. We performed a 2 × 2 factorial design experiment on a total of 40 sheep ovaries, divided into four groups (n = 10 ovaries per group): (i) directional freezing of whole ovary (DFwo); (ii) directional freezing of ovarian fragments (DFof); (iii) conventional freezing of whole ovary (CFwo); (iv) conventional freezing of ovarian fragments (CFof). An additional eight ovaries were used as fresh controls. Ewe ovaries were randomly assigned to one of the experimental groups and frozen accordingly. Upon thawing, ovarian tissue was examined morphologically and cultured in vitro for 7 days. Samples were analyzed for cell proliferation and apoptosis, for DNA damage and repair activity, and for the presence of a panel of heat shock proteins (HSPs) by immunohistochemistry. Most studied parameters were significantly improved (P < 0.05) in all samples cryopreserved with directional compared with slow freezing. The proportion of primordial follicles, which developed to the primary stage in whole ovaries (53 ± 1.7%) and in ovarian fragments (44 ± 1.8%) cryopreserved with directional freezing, was greater than with slow frozen whole ovaries (6 ± 0.5%, P = 0.001) or fragments (32 ± 1.5%, P = 0.004). After 7 days of culture, cell proliferation in DFwo (28 ± 0.73%) was the highest of all groups (P < 0.05) followed by DFof (23 ± 0.81%), CFof (20 ± 0.79%) and CFwo (9 ± 0.85%). Directional freezing also resulted in a better preservation of the cell capacity to repair DNA damage compared with slow freezing both in whole ovaries and ovarian fragments. Apoptosis and HSP protein levels were significantly increased only in the CFwo group. Direct comparison demonstrated that, overall, DFwo had better parameters than DFof and was no different from the fresh controls. The study is limited to an in vitro evaluation and uses sheep ovaries, which are smaller than human ovaries and therefore may withstand the procedures better. Improved integrity of ovarian morphology may translate to improved outcomes after transplantation. Alternatively, the particularly good preservation of whole ovaries suggests they could provide a source of ovarian follicles for in vitro culture in those cases when the presence of malignant cells poses a substantial risk for the patient. Supported by: Associazione Italiana per la Ricerca sul Cancro (AIRC) IG 10376, Carraresi Foundation and by Legge 7 Regione Autonoma Sardegna (R.A.S). There are no conflicts of interest.

49 CFR 1039.10 - Exemption of agricultural commodities except grain, soybeans, and sunflower seeds.

Code of Federal Regulations, 2010 CFR

2010-10-01

... other marine products. 20-11 Fresh meat. 20-15 Fresh dressed poultry. 20-17 Processed poultry. 20-21.... 20-999-76 Freeze-dried poultry. 20-999-77 Freeze-dried meat. 20-999-78 Freeze-dried salad ingredients...

Effect of Freezing on the Level of Contaminants in Uncontrolled Hazardous Waste Sites. Part 1. Literature Review.

DTIC Science & Technology

1986-07-01

pure water. Dissolved ions in the soil solution lower the freezing point; this is called freezing point depression. Many of the early studies of...them in the remaining soil solution . The temperature and concentration of this solution affect the chemical reactions and the forms of ions in...in the soil solution freezes, more concentrated "% solutes will be present in soil solution . 3. Water will travel even in frozen soils and sediments

Preparation of freezing quantum state for quantum coherence

NASA Astrophysics Data System (ADS)

Yang, Lian-Wu; Man, Zhong-Xiao; Zhang, Ying-Jie; Han, Feng; Du, Shao-jiang; Xia, Yun-Jie

2018-06-01

We provide a method to prepare the freezing quantum state for quantum coherence via unitary operations. The initial product state consists of the control qubit and target qubit; when it satisfies certain conditions, the initial product state converts into the particular Bell diagonal state under the unitary operations, which have the property of freezing of quantum coherence under quantum channels. We calculate the frozen quantum coherence and corresponding quantum correlations, and find that the quantities are determined by the control qubit only when the freezing phenomena occur.

New High-Performance Droplet Freezing Assay (HP-DFA) for the Analysis of Ice Nuclei with Complex Composition

NASA Astrophysics Data System (ADS)

Kunert, Anna Theresa; Scheel, Jan Frederik; Helleis, Frank; Klimach, Thomas; Pöschl, Ulrich; Fröhlich-Nowoisky, Janine

2016-04-01

Freezing of water above homogeneous freezing is catalyzed by ice nucleation active (INA) particles called ice nuclei (IN), which can be of various inorganic or biological origin. The freezing temperatures reach up to -1 °C for some biological samples and are dependent on the chemical composition of the IN. The standard method to analyze IN in solution is the droplet freezing assay (DFA) established by Gabor Vali in 1970. Several modifications and improvements were already made within the last decades, but they are still limited by either small droplet numbers, large droplet volumes or inadequate separation of the single droplets resulting in mutual interferences and therefore improper measurements. The probability that miscellaneous IN are concentrated together in one droplet increases with the volume of the droplet, which can be described by the Poisson distribution. At a given concentration, the partition of a droplet into several smaller droplets leads to finely dispersed IN resulting in better statistics and therefore in a better resolution of the nucleation spectrum. We designed a new customized high-performance droplet freezing assay (HP-DFA), which represents an upgrade of the previously existing DFAs in terms of temperature range and statistics. The necessity of observing freezing events at temperatures lower than homogeneous freezing due to freezing point depression, requires high-performance thermostats combined with an optimal insulation. Furthermore, we developed a cooling setup, which allows both huge and tiny temperature changes within a very short period of time. Besides that, the new DFA provides the analysis of more than 750 droplets per run with a small droplet volume of 5 μL. This enables a fast and more precise analysis of biological samples with complex IN composition as well as better statistics for every sample at the same time.

The impact of freeze-drying infant fecal samples on measures of their bacterial community profiles and milk-derived oligosaccharide content.

PubMed

Lewis, Zachery T; Davis, Jasmine C C; Smilowitz, Jennifer T; German, J Bruce; Lebrilla, Carlito B; Mills, David A

2016-01-01

Infant fecal samples are commonly studied to investigate the impacts of breastfeeding on the development of the microbiota and subsequent health effects. Comparisons of infants living in different geographic regions and environmental contexts are needed to aid our understanding of evolutionarily-selected milk adaptations. However, the preservation of fecal samples from individuals in remote locales until they can be processed can be a challenge. Freeze-drying (lyophilization) offers a cost-effective way to preserve some biological samples for transport and analysis at a later date. Currently, it is unknown what, if any, biases are introduced into various analyses by the freeze-drying process. Here, we investigated how freeze-drying affected analysis of two relevant and intertwined aspects of infant fecal samples, marker gene amplicon sequencing of the bacterial community and the fecal oligosaccharide profile (undigested human milk oligosaccharides). No differences were discovered between the fecal oligosaccharide profiles of wet and freeze-dried samples. The marker gene sequencing data showed an increase in proportional representation of Bacteriodes and a decrease in detection of bifidobacteria and members of class Bacilli after freeze-drying. This sample treatment bias may possibly be related to the cell morphology of these different taxa (Gram status). However, these effects did not overwhelm the natural variation among individuals, as the community data still strongly grouped by subject and not by freeze-drying status. We also found that compensating for sample concentration during freeze-drying, while not necessary, was also not detrimental. Freeze-drying may therefore be an acceptable method of sample preservation and mass reduction for some studies of microbial ecology and milk glycan analysis.

Transcriptional and metabolomic analysis of Ascophyllum nodosum mediated freezing tolerance in Arabidopsis thaliana

PubMed Central

2012-01-01

Background We have previously shown that lipophilic components (LPC) of the brown seaweed Ascophyllum nodosum (ANE) improved freezing tolerance in Arabidopsis thaliana. However, the mechanism(s) of this induced freezing stress tolerance is largely unknown. Here, we investigated LPC induced changes in the transcriptome and metabolome of A. thaliana undergoing freezing stress. Results Gene expression studies revealed that the accumulation of proline was mediated by an increase in the expression of the proline synthesis genes P5CS1 and P5CS2 and a marginal reduction in the expression of the proline dehydrogenase (ProDH) gene. Moreover, LPC application significantly increased the concentration of total soluble sugars in the cytosol in response to freezing stress. Arabidopsis sfr4 mutant plants, defective in the accumulation of free sugars, treated with LPC, exhibited freezing sensitivity similar to that of untreated controls. The 1H NMR metabolite profile of LPC-treated Arabidopsis plants exposed to freezing stress revealed a spectrum dominated by chemical shifts (δ) representing soluble sugars, sugar alcohols, organic acids and lipophilic components like fatty acids, as compared to control plants. Additionally, 2D NMR spectra suggested an increase in the degree of unsaturation of fatty acids in LPC treated plants under freezing stress. These results were supported by global transcriptome analysis. Transcriptome analysis revealed that LPC treatment altered the expression of 1113 genes (5%) in comparison with untreated plants. A total of 463 genes (2%) were up regulated while 650 genes (3%) were down regulated. Conclusion Taken together, the results of the experiments presented in this paper provide evidence to support LPC mediated freezing tolerance enhancement through a combination of the priming of plants for the increased accumulation of osmoprotectants and alteration of cellular fatty acid composition. PMID:23171218

Experimental and numerical study on frost heave of saturated rock under uniform freezing conditions

NASA Astrophysics Data System (ADS)

Lv, Zhitao; Xia, Caichu; Li, Qiang

2018-04-01

A series of freezing experiments are conducted on saturated sandstone and mortar specimens to investigate the frost heave of saturated rock under uniform freezing conditions. The experimental results show that the frost heave of saturated rock is isotropic under uniform freezing conditions. During the freezing process, three stages are observed in the curves of variation of total frost heaving strain versus time: the thermal contraction stage, the frost heaving stage and the steady stage. Moreover, the amount of final stable frost heave first increases and then decreases with decrease in freezing temperature, and the maximum final stable frost heave occurs at different freezing temperature in saturated sandstone and mortar. Furthermore, a coupled thermal-mechanical (TM) model of frost heave of saturated rock is proposed in which a constraint coefficient \\zeta is used to consider the susceptibility of the internal rock grain structure to the expansion of pore ice. Then, numerical simulations are implemented with COMSOL to solve the governing equations of the TM model. Comparisons of the numerical results with the experimental results are performed to demonstrate the reliability of the model. The influences of elastic modulus and porosity on frost heave are also investigated, and the results show that the total frost heaving strain decreases non-linearly with increasing elastic modulus, and the decrease is significant when the elastic modulus is less than 3000 MPa, or approximately five times the elastic modulus of ice. In addition, the total frost heaving strain increases linearly with increasing porosity. Finally, an empirical equation between total frost heaving strain and freezing temperature is proposed and the equation well describes the variation of total frost heaving strain with freezing temperature.

Overexpression of the calcineurin target CRZ1 provides freeze tolerance and enhances the fermentative capacity of baker's yeast.

PubMed

Panadero, Joaquín; Hernández-López, Maria José; Prieto, José Antonio; Randez-Gil, Francisca

2007-08-01

Recent years have shown a huge growth in the market of industrial baker's yeasts (Saccharomyces cerevisiae), with the need for strains affording better performance in prefrozen dough. Evidence suggests that during the freezing process, cells can suffer biochemical damage caused by osmotic stress. Nevertheless, the involvement of ion-responsive transcriptional factors and pathways in conferring freeze resistance has not yet been examined. Here, we have investigated the role of the salt-responsive calcineurin-Crz1p pathway in mediating tolerance to freezing by industrial baker's yeast. Overexpression of CRZ1 in the industrial HS13 strain increased both salt and freeze tolerance and improved the leavening ability of baker's yeast in high-sugar dough. Moreover, engineered cells were able to produce more gas during fermentation of prefrozen dough than the parental strain. Similar effects were observed for overexpression of TdCRZ1, the homologue to CRZ1 in Torulaspora delbrueckii, suggesting that expression of calcineurin-Crz1p target genes can alleviate the harmful effects of ionic stress during freezing. However, overexpression of STZ and FTZ, two unrelated Arabidopsis thaliana genes encoding Cys(2)/His(2)-type zinc finger proteins, also conferred freeze resistance in yeast. Furthermore, experiments with Deltacnb1 and Deltacrz1 mutants failed to show a freeze-sensitive phenotype, even in cells pretreated with NaCl. Overall, our results demonstrate that overexpression of CRZ1 has the potential to be a useful tool for increasing freeze tolerance and fermentative capacity in industrial strains. However, these effects do not appear to be mediated through activation of known salt-responding pathways.

Effect of Repeated Freeze-Thaw Cycles on Beef Quality and Safety

PubMed Central

Rahman, Mohammad Hafizur; Hossain, Mohammad Mujaffar; Rahman, Syed Mohammad Ehsanur; Hashem, Mohammad Abul

2014-01-01

The objectives of this study were to know the effect of repeated freeze-thaw cycles of beef on the sensory, physicochemical quality and microbiological assessment. The effects of three successive freeze-thaw cycles on beef forelimb were investigated comparing with unfrozen fresh beef for 75 d by keeping at −20±1℃. The freeze-thaw cycles were subjected to three thawing methods and carried out to know the best one. As the number of freeze-thaw cycles increased color and odor declined significantly before cook within the cycles and tenderness, overall acceptability also declined among the cycles after cook by thawing methods. The thawing loss increased and dripping loss decreased significantly (p<0.05). Water holding capacity (WHC) increased (p<0.05) until two cycles and then decreased. Cooking loss increased in cycle 1 and 3, but decreased in cycle 2. pH decreased significantly (p<0.05) among the cycles. Moreover, drip loss, cooking loss and WHC were affected (p<0.05) by thawing methods within the cycles. 2-Thiobarbituric acid (TBARS) value increased (p<0.05) gradually within the cycles and among the cycles by thawing methods. Total viable bacteria, total coliform and total yeast-mould count decreased significantly (p<0.05) within and among the cycles in comparison to the initial count in repeated freeze-thaw cycles. As a result, repeated freeze-thaw cycles affected the sensory, physicochemical and microbiological qua- lity of beef, causing the deterioration of beef quality, but improved the microbiological quality. Although repeated freeze-thaw cycles did not affect much on beef quality and safety but it may be concluded that repeated freeze and thaw should be minimized in terms of beef color for commercial value and WHC and tenderness/juiciness for eating quality. PMID:26761286

Cryoprotectants and Extreme Freeze Tolerance in a Subarctic Population of the Wood Frog

PubMed Central

Costanzo, Jon P.; Reynolds, Alice M.; do Amaral, M. Clara F.; Rosendale, Andrew J.; Lee, Richard E.

2015-01-01

Wood frogs (Rana sylvatica) exhibit marked geographic variation in freeze tolerance, with subarctic populations tolerating experimental freezing to temperatures at least 10-13 degrees Celsius below the lethal limits for conspecifics from more temperate locales. We determined how seasonal responses enhance the cryoprotectant system in these northern frogs, and also investigated their physiological responses to somatic freezing at extreme temperatures. Alaskan frogs collected in late summer had plasma urea levels near 10 μmol ml-1, but this level rose during preparation for winter to 85.5 ± 2.9 μmol ml-1 (mean ± SEM) in frogs that remained fully hydrated, and to 186.9 ± 12.4 μmol ml-1 in frogs held under a restricted moisture regime. An osmolality gap indicated that the plasma of winter-conditioned frogs contained an as yet unidentified osmolyte(s) that contributed about 75 mOsmol kg-1 to total osmotic pressure. Experimental freezing to –8°C, either directly or following three cycles of freezing/thawing between –4 and 0°C, or –16°C increased the liver’s synthesis of glucose and, to a lesser extent, urea. Concomitantly, organs shed up to one-half (skeletal muscle) or two-thirds (liver) of their water, with cryoprotectant in the remaining fluid reaching concentrations as high as 0.2 and 2.1 M, respectively. Freeze/thaw cycling, which was readily survived by winter-conditioned frogs, greatly increased hepatic glycogenolysis and delivery of glucose (but not urea) to skeletal muscle. We conclude that cryoprotectant accrual in anticipation of and in response to freezing have been greatly enhanced and contribute to extreme freeze tolerance in northern R. sylvatica. PMID:25688861

Inactivation kinetics of Vibrio vulnificus in phosphate-buffered saline at different freezing and storage temperatures and times.

PubMed

Seminario, Diana M; Balaban, Murat O; Rodrick, Gary

2011-03-01

Vibrio vulnificus (Vv) is a pathogen that can be found in raw oysters. Freezing can reduce Vv and increase the shelf life of oysters. The objective of this study was to develop predictive inactivation kinetic models for pure cultures of Vv at different frozen storage temperatures and times. Vv was diluted in phosphate-buffered saline (PBS) to obtain about 10(7) CFU/mL. Samples were frozen at -10, -35, and -80 °C (different freezing rates), and stored at different temperatures. Survival of Vv was followed after freezing and storage at -10 °C (0, 3, 6, and 9 d) and at -35 and -80 °C (every week for 6 wk). For every treatment, time-temperature data was obtained using thermocouples in blank vials. Predictive models were developed using first-order, Weibull and Peleg inactivation kinetics. Different freezing temperatures did not significantly (α = 0.05) affect survival of Vv immediately after freezing. The combined effect of freezing and 1 wk frozen storage resulted in 1.5, 2.6, and 4.9 log10 reductions for samples stored at -80, -35, and -10 °C, respectively. Storage temperature was the critical parameter in survival of Vv. A modified Weibull model successfully predicted Vv survival during frozen storage: log10 Nt = log 10No - 1.22 - ([t/10{-1.163-0.0466T}][0.00025T(2) + 0.049325]). N(o) and N(t) are initial and time t (d) survival counts, T is frozen storage temperature, Celsius degree. Vibrio vulnificus can be inactivated by freezing. Models to predict survival of V. vulnificus at different freezing temperatures and times were developed. This is the first step towards the prediction of V. vulnificus related safety of frozen oysters.

Three keys to the radiation of angiosperms into freezing environments.

PubMed

Zanne, Amy E; Tank, David C; Cornwell, William K; Eastman, Jonathan M; Smith, Stephen A; FitzJohn, Richard G; McGlinn, Daniel J; O'Meara, Brian C; Moles, Angela T; Reich, Peter B; Royer, Dana L; Soltis, Douglas E; Stevens, Peter F; Westoby, Mark; Wright, Ian J; Aarssen, Lonnie; Bertin, Robert I; Calaminus, Andre; Govaerts, Rafaël; Hemmings, Frank; Leishman, Michelle R; Oleksyn, Jacek; Soltis, Pamela S; Swenson, Nathan G; Warman, Laura; Beaulieu, Jeremy M

2014-02-06

Early flowering plants are thought to have been woody species restricted to warm habitats. This lineage has since radiated into almost every climate, with manifold growth forms. As angiosperms spread and climate changed, they evolved mechanisms to cope with episodic freezing. To explore the evolution of traits underpinning the ability to persist in freezing conditions, we assembled a large species-level database of growth habit (woody or herbaceous; 49,064 species), as well as leaf phenology (evergreen or deciduous), diameter of hydraulic conduits (that is, xylem vessels and tracheids) and climate occupancies (exposure to freezing). To model the evolution of species' traits and climate occupancies, we combined these data with an unparalleled dated molecular phylogeny (32,223 species) for land plants. Here we show that woody clades successfully moved into freezing-prone environments by either possessing transport networks of small safe conduits and/or shutting down hydraulic function by dropping leaves during freezing. Herbaceous species largely avoided freezing periods by senescing cheaply constructed aboveground tissue. Growth habit has long been considered labile, but we find that growth habit was less labile than climate occupancy. Additionally, freezing environments were largely filled by lineages that had already become herbs or, when remaining woody, already had small conduits (that is, the trait evolved before the climate occupancy). By contrast, most deciduous woody lineages had an evolutionary shift to seasonally shedding their leaves only after exposure to freezing (that is, the climate occupancy evolved before the trait). For angiosperms to inhabit novel cold environments they had to gain new structural and functional trait solutions; our results suggest that many of these solutions were probably acquired before their foray into the cold.

Freezing and drying effects on potential plant contributions to phosphorus in runoff.

PubMed

Roberson, Tiffany; Bundy, Larry G; Andraski, Todd W

2007-01-01

Phosphorus (P) in runoff from landscapes can promote eutrophication of natural waters. Soluble P released from plant material can contribute significant amounts of P to runoff particularly after plant freezing or drying. This study was conducted to evaluate P losses from alfalfa or grass after freezing or drying as potential contributors to runoff P. Alfalfa (Medicago sativa L.) and grass (principally, Agropyron repens L.) plant samples were subjected to freezing and drying treatments to determine P release. Simulated rainfall runoff and natural runoff from established alfalfa fields and a grass waterway were collected to study P contributions from plant tissue to runoff. The effects of freezing and drying on P released from plant tissue were simulated by a herbicide treatment in selected experiments. Soluble reactive P (SP) extracted from alfalfa and grass samples was markedly increased by freezing or drying. In general, SP extracted from plant samples increased in the order fresh < frozen < frozen/thawed < dried, and averaged 1, 8, 14, and 26% of total P in alfalfa, respectively. Soluble reactive P extracted from alfalfa after freezing or drying increased with increasing soil test P (r(2) = 0.64 to 0.68), suggesting that excessive soil P levels increased the risk of plant P contributions to runoff losses. In simulated rainfall studies, paraquat (1,1'-dimethyl-4, 4''-bipyridinium ion) treatment of alfalfa increased P losses in runoff, and results suggested that this treatment simulated the effects of drying on plant P loss. In contrast to the simulated rainfall results, natural runoff studies over 2 yr did not show higher runoff P losses that could be attributed to P from alfalfa. Actual P losses likely depend on the timing and extent of plant freezing and drying and of precipitation events after freezing.

Identification, analysis and monitoring of risks of freezing affecting aircraft flying over the Guadarrama Mountains (Spain)

NASA Astrophysics Data System (ADS)

Fernández-González, Sergio; Sánchez, José Luis; Gascón, Estíbaliz; Merino, Andrés; Hermida, Lucía; López, Laura; Marcos, José Luis; García-Ortega, Eduardo

2014-05-01

Freezing is one of the main causes of aircraft accidents registered over the last few decades. This means it is very important to be able to predict this situation so that aircraft can change their routes to avoid freezing risk areas. Also, by using satellites it is possible to observe changes in the horizontal and vertical extension of cloud cover likely to cause freezing in real time as well as microphysical changes in the clouds. The METEOSAT Second Generation (MSG) makes it possible to create different red-green-blue (RGB) compositions that provide a large amount of information associated with the microphysics of clouds, in order to identify super-cooled water clouds that pose a high risk of freezing to aircraft. During the winter of 2011/12 in the Guadarrama Mountains, in the centre of the Iberian Peninsula, a series of scientific flights (conducted by INTA) were organised in order to study the cloud systems that affected this region during the winter. On the flight of the 1st of February 2012, the aircraft was affected by freezing after crossing over a mountain ridge with supercooled large drops (SLD). Although freezing was not expected during that day's flight, the orography caused a series of mesoscale factors that led to the appearance of localised freezing conditions. By analysing this case, we have been able to conclude that the use of satellite images makes it possible to monitor the risk of freezing, especially under specific mesoscale circumstances. Acknowledgements S. Fernández-González acknowledges the grant supported from the FPU program (AP 2010-2093). This study was supported by the following grants: GRANIMETRO (CGL2010-15930); MICROMETEO (IPT-310000-2010-22). The authors would like to thank the INTA for its scientific flights.

Emerging freeze-drying process development and scale-up issues.

PubMed

Patel, Sajal Manubhai; Pikal, Michael J

2011-03-01

Although several guidelines do exist for freeze-drying process development and scale-up, there are still a number of issues that require additional attention. The objective of this review article is to discuss some emerging process development and scale-up issue with emphasis on effect of load condition and freeze-drying in novel container systems such as syringes, Lyoguard trays, ampoules, and 96-well plates. Understanding the heat and mass transfer under different load conditions and for freeze-drying in these novel container systems will help in developing a robust freeze-drying process which is also easier to scale-up. Further research and development needs in these emerging areas have also been addressed. © 2011 American Association of Pharmaceutical Scientists

7 CFR 305.7 - Quick freeze treatment requirements.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 5 2012-01-01 2012-01-01 false Quick freeze treatment requirements. 305.7 Section 305.7 Agriculture Regulations of the Department of Agriculture (Continued) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE PHYTOSANITARY TREATMENTS § 305.7 Quick freeze treatment...

Development and demonstration of a freezing drizzle algorithm for roadway environmental sensing Systems.

DOT National Transportation Integrated Search

2012-10-01

The primary goal of this project is to demonstrate the accuracy and utility of a freezing drizzle algorithm that can be implemented on roadway environmental sensing systems (ESSs). : The types of problems related to the occurrence of freezing precipi...

Development of a Method to Produce Freeze-Dried Cubes from 3 Pacific Salmon Species

USDA-ARS?s Scientific Manuscript database

Freeze-dried boneless skinless cubes of pink (Oncorhynchus gorbuscha), sockeye (Oncorhynchus nerka), and chum (Oncorhynchus keta) salmon were prepared and physical properties evaluated. To minimize freeze-drying time, the kinetics of dehydration and processing yields were investigated. The physical ...

Experimental research of "microcable in a microconduct" system stability to effect of freezing water

NASA Astrophysics Data System (ADS)

Andreev, Vladimir A.; Burdin, Vladimir A.; Nikulina, Tatiana G.; Alekhin, Ivan N.; Gavryushin, Sergey A.; Nikulin, Aleksey G.; Praporshchikov, Denis E.

2011-12-01

Results of experimental researches of "optical microcable in a microduct" system stability to effect of freezing water are presented. It is shown this system is steadier to water freezing in comparison to lighten optical cable in protective polymer tube.

Freeze-drying of lactic acid bacteria.

PubMed

Fonseca, Fernanda; Cenard, Stéphanie; Passot, Stéphanie

2015-01-01

Lactic acid bacteria are of great importance for the food and biotechnology industry. They are widely used as starters for manufacturing food (e.g., yogurt, cheese, fermented meats, and vegetables) and probiotic products, as well as for green chemistry applications. Freeze-drying or lyophilization is a convenient method for preservation of bacteria. By reducing water activity to values below 0.2, it allows long-term storage and low-cost distribution at suprazero temperatures, while minimizing losses in viability and functionality. Stabilization of bacteria via freeze-drying starts with the addition of a protectant solution to the bacterial suspension. Freeze-drying includes three steps, namely, (1) freezing of the concentrated and protected cell suspension, (2) primary drying to remove ice by sublimation, and (3) secondary drying to remove unfrozen water by desorption. In this chapter we describe a method for freeze-drying of lactic acid bacteria at a pilot scale, thus allowing control of the process parameters for maximal survival and functionality recovery.

'Banking time': egg freezing and the negotiation of future fertility.

PubMed

Waldby, Catherine

2015-01-01

This paper examines the relatively recent practice of non-medical egg freezing, in which women bank their eggs for later use in conceiving a child. Non-medical egg freezing has only been available for about the last five years, as new vitrification techniques have made the success rates for actual conception more reliable than the earlier method of slow freezing. I draw on interviews with both clinicians and women who have banked their eggs to consider how this novel practice articulates with broader issues about the relationship between sexuality, reproduction and the political economy of household formation. Non-medical egg-freezing provides a technical solution to a number of different problems women face with regard to the elongation of the life course, the extension of education, the cost of household establishment and the iterative nature of relationship formation, thematised by the ubiquity of internet dating among the interviewees. I focus on the ways women used egg freezing to manage and reconcile different forms of time.

Cryopreservation of day 2-3 embryos by vitrification yields better outcome than slow freezing.

PubMed

Levron, Jacob; Leibovitz, Oshrit; Brengauz, Masha; Gitman, Hila; Yerushalmi, Gil M; Katorza, Eldad; Gat, Itai; Elizur, Shai E

2014-03-01

To compare the outcome of vitrification versus slow freezing cryopreservation for cleavage stage day 2-3 embryos. A retrospective observational study. All thawed embryos assisted reproduction cycles between January 2010 and December 2012 at a single IVF laboratory of a Tertiary Medical Center. Five hundred and thirty-nine cycles of day 2-3 thawed embryos. In 327 of the thawed cycles, the embryos were vitrified and in 212 of the cycles the embryos were derived from slow freezing embryos. Embryo survival rate, blastomere surviving rate and pregnancy rate. Embryo survival rate was significantly higher after vitrification compared with slow freezing (81.6%, 647/793 versus 70.0%, 393/562 embryos, p < 0.0001). The clinical pregnancy rate per ET was significantly higher following vitrification compared to slow freezing, 20.0%, 63/314 versus 11.9%, 23/193, respectively (p = 0.02). Vitrification of day 2-3 cleavage stage embryos yields better cycle outcome in all the parameters compared to slow freezing.

Dynamical freeze-out criterion in a hydrodynamical description of Au + Au collisions at √{sNN}=200 GeV and Pb + Pb collisions at √{sNN}=2760 GeV

NASA Astrophysics Data System (ADS)

Ahmad, Saeed; Holopainen, Hannu; Huovinen, Pasi

2017-05-01

In hydrodynamical modeling of ultrarelativistic heavy-ion collisions, the freeze-out is typically assumed to take place at a surface of constant temperature or energy density. A more physical approach is to assume that freeze-out takes place at a surface of constant Knudsen number. We evaluate the Knudsen number as a ratio of the expansion rate of the system to the pion-scattering rate and apply the constant Knudsen number freeze-out criterion to the ideal hydrodynamical description of heavy-ion collisions at the Relativistic Heavy Ion Collider at BNL (√{sNN}=200 GeV) and the Large Hadron Collider (√{sNN}=2760 GeV) energies. We see that once the numerical values of freeze-out temperature and freeze-out Knudsen number are chosen to produce similar pT distributions, the elliptic and triangular anisotropies are similar too, in both event-by-event and averaged initial state calculations.

The initial freezing point temperature of beef rises with the rise in pH: a short communication.

PubMed

Farouk, M M; Kemp, R M; Cartwright, S; North, M

2013-05-01

This study tested the hypothesis that the initial freezing point temperature of meat is affected by pH. Sixty four bovine M. longissimus thoracis et lumborum were classified into two ultimate pH groups: low (<5.8) and high pH (>6.2) and their cooling and freezing point temperatures were determined. The initial freezing temperatures for beef ranged from -0.9 to -1.5°C (∆=0.6°C) with the higher and lower temperatures associated with high and low ultimate pH respectively. There was a significant correlation (r=+0.73, P<0.01) between beef pH and freezing point temperature in the present study. The outcome of this study has implications for the meat industry where evidence of freezing (ice formation) in a shipment as a result of high pH meat could result in a container load of valuable chilled product being downgraded to a lower value frozen product. Copyright © 2013 Elsevier Ltd. All rights reserved.

Freezing, thawing and aging effects on beef tenderness from Bos indicus and Bos taurus cattle.

PubMed

Aroeira, Carolina N; Torres Filho, Robledo A; Fontes, Paulo Rogério; Gomide, Lúcio Alberto M; Ramos, Alcinéia L S; Ladeira, Márcio M; Ramos, Eduardo M

2016-06-01

The objective of this study was to determine the effects of freezing prior to aging on the meat tenderness of young Nellore and Aberdeen Angus bulls. Samples of the longissimus thoracis muscle were submitted to two treatments: conventional aging and freezing (-20°C for 40 days) followed by thawing and aging periods. The meats were evaluated after 0, 7, 14 and 21 aging days (1°C). Freezing increased (P<0.05) purge, cooking loss and total exudate loss throughout aging. Nellore meats had greater total exudate loss and shorter sarcomere lengths (P<0.05). Freezing increased proteolysis during aging in the meats of both breeds, but reduced shear force was found (P<0.05) only in Aberdeen Angus meats and only at time zero. These results suggest that the meat tenderizing process by freezing prior to aging may contribute to meat tenderness in the first weeks of aging, but it is dependent on the animal breed. Copyright © 2016 Elsevier Ltd. All rights reserved.

Dispersion Stability of O/W Emulsions with Different Oil Contents Under Various Freezing and Thawing Conditions.

PubMed

Katsuki, Kazutaka; Miyagawa, Yayoi; Nakagawa, Kyuya; Adachi, Shuji

2017-07-01

Freezing and thawing of oil-in-water (O/W) emulsion-type foods bring about oil-water separation and deterioration; hence, the effects of freezing and thawing conditions on the destabilization of O/W emulsions were examined. The freezing rate and thawing temperature hardly affected the stability of the O/W emulsion. O/W emulsions having different oil fractions were stored at temperatures ranging from -30 to -20 °C and then thawed. The stability after thawing depended on the storage temperature, irrespective of the oil fraction of the emulsion. A good correlation was found between the time at which the stability began to decrease and the time taken for the oil to crystalize. These results indicated that the dominant cause for the destabilization of the O/W emulsion during freezing and thawing is the crystallization of the oil phase and that the effects of the freezing and thawing rates on the stability are insignificant. © 2017 Institute of Food Technologists®.

Effect of freezing prior to aging on myoglobin redox forms and CIE color of beef from Nellore and Aberdeen Angus cattle.

PubMed

Aroeira, Carolina Naves; de Almeida Torres Filho, Robledo; Fontes, Paulo Rogério; de Lemos Souza Ramos, Alcinéia; de Miranda Gomide, Lúcio Alberto; Ladeira, Márcio Machado; Ramos, Eduardo Mendes

2017-03-01

The objective of this study was to evaluate the effect of freezing prior to wet aging on the color of Nellore and Aberdeen Angus cattle meat. Samples of the Longissimus thoracis muscle were subjected to two treatments: conventional aging (0, 7, 14 and 21days); and freezing (-20°C for 40days) followed by thawing and aging. Freezing promoted (P<0.05) formation of metmyoglobin during aging, especially in Nellore beef. Frozen meats showed (P<0.05) lower lightness (L*) values and higher redness (a*), chroma (C*) and hue angle (h*) values at the first day of storage, deteriorating quickly with aging time. The color of the Nellore meat was less (P<0.05) stable to freezing, being lighter, yellower and less red than Angus meat. The results suggest that color stability in vacuum-packed beef is reduced by freezing prior to aging and that reduction depends on the animal breed. Copyright © 2016 Elsevier Ltd. All rights reserved.

Influence of waxy rice flour substitution for wheat flour on characteristics of batter and freeze-thawed cake.

PubMed

Jongsutjarittam, Nisachon; Charoenrein, Sanguansri

2013-09-12

This study aimed to improve the freeze-thawed cake properties by10-20% waxy rice flour (WRF) substitution for wheat flour (WF). Viscosity of WRF-substituted batters was lower; consequently, trapped air was less uniformly distributed than WF batter. After five freeze-thaw cycles, firmness and enthalpy of melting retrograded amylopectin of WF- and WRF-substituted cakes increased and the matrix surrounding the air pores from SEM images was denser than in fresh-baked cakes. Sensory evaluation showed an increase in firmness and a decrease in firmness acceptability of freeze-thawed cakes. However, freeze-thawed cake with WRF substitution had significantly less firmness, less dense matrix and more acceptability than WF cake. This could have been due to a low amylose content of WRF and the spread of ruptured waxy rice starch granules around swollen wheat starch granules as observed by CLSM. Thus, WRF could be used for WF substitution to improve the firmness in freeze-thawed cake. Copyright © 2013 Elsevier Ltd. All rights reserved.

Fundamentals of freeze-drying.

PubMed

Nail, Steven L; Jiang, Shan; Chongprasert, Suchart; Knopp, Shawn A

2002-01-01

Given the increasing importance of reducing development time for new pharmaceutical products, formulation and process development scientists must continually look for ways to "work smarter, not harder." Within the product development arena, this means reducing the amount of trial and error empiricism in arriving at a formulation and identification of processing conditions which will result in a quality final dosage form. Characterization of the freezing behavior of the intended formulation is necessary for developing processing conditions which will result in the shortest drying time while maintaining all critical quality attributes of the freeze-dried product. Analysis of frozen systems was discussed in detail, particularly with respect to the glass transition as the physical event underlying collapse during freeze-drying, eutectic mixture formation, and crystallization events upon warming of frozen systems. Experiments to determine how freezing and freeze-drying behavior is affected by changes in the composition of the formulation are often useful in establishing the "robustness" of a formulation. It is not uncommon for seemingly subtle changes in composition of the formulation, such as a change in formulation pH, buffer salt, drug concentration, or an additional excipient, to result in striking differences in freezing and freeze-drying behavior. With regard to selecting a formulation, it is wise to keep the formulation as simple as possible. If a buffer is needed, a minimum concentration should be used. The same principle applies to added salts: If used at all, the concentration should be kept to a minimum. For many proteins a combination of an amorphous excipient, such as a disaccharide, and a crystallizing excipient, such as glycine, will result in a suitable combination of chemical stability and physical stability of the freeze-dried solid. Concepts of heat and mass transfer are valuable in rational design of processing conditions. Heat transfer by conduction--the dominant mechanism of heat transfer in freeze-drying--is inefficient at the pressures used in freeze-drying. Steps should be taken to improve the thermal contact between the product and the shelf of the freeze dryer, such as eliminating metal trays from the drying process. Quantitation of the heat transfer coefficient for the geometry used is a useful way of assessing the impact of changes in the system such as elimination of product trays and changes in the vial. Because heat transfer by conduction through the vapor increases with increasing pressure, the commonly held point of view that "the lower the pressure, the better" is not true with respect to process efficiency. The optimum pressure for a given product is a function of the temperature at which freeze-drying is carried out, and lower pressures are needed at low product temperatures. The controlling resistance to mass transfer is almost always the resistance of the partially dried solids above the submination interface. This resistance can be minimized by avoiding fill volumes of more than about half the volume of the container. The development scientist should also recognize that very high concentrations of solute may not be appropriate for optimum freeze-drying, particularly if the resistance of the dried product layer increases sharply with concentration. Although the last 10 years has seen the publication of a significant body of literature of great value in allowing development scientists and engineers to "work smarter," there is still much work needed in both the science and the technology of freeze-drying. Scientific development is needed for improving analytical methodology for characterization of frozen systems and freeze-dried solids. A better understanding of the relationship between molecular mobility and reactivity is needed to allow accurate prediction of product stability at the intended storage temperature based on accelerated stability at higher temperatures. This requires that the temperature dependence of glass transition-associated mobility, particularly at temperatures below the glass transition, be studied in greater depth. The relevance of the concept of strong and fragile glasses to frozen systems and freeze-dried solids has only begun to be explored. The list of pharmaceutically acceptable protective solutes is very short, and more imagination--and work--is needed in order to develop pharmaceutically acceptable alternative stabilizers. There is a need for technology development in process monitoring, particularly in developing a way to measure the status of the product during freezing and freeze-drying without placing temperature measurement probes in individual vials of product. The current practice of placing thermocouples in vials is uncertain with respect to reliability of the data, inconsistent with elimination of personnel in close proximity to open vials of product in an aseptic environment, and incompatible with technology for automatic material handling in freeze-drying. In addition, a method for controlling the degree of supercooling during freezing would allow better control of freezing rate and would, in many cases, result in more consistent product quality.

Freeze-dried dog sperm: Dynamics of DNA integrity.

PubMed

Olaciregui, M; Luño, V; Gonzalez, N; De Blas, I; Gil, L

2015-10-01

Freeze-drying (FD) has been proposed as an alternative method to preserve spermatozoa. During the FD procedure, sperm DNA might become damaged by both freezing and drying stresses caused by the endonucleases, the oxidative stress and the storage conditions. We examined the DNA integrity of dog sperm freeze-dried with two kinds of chelating agents in FD buffers and storage at two different temperatures. Ejaculated sperm from four dogs were suspended in basic medium (10 mM Tris-HCl buffer+50 mM NaCl) supplemented with 50 mM EGTA or with 50 mM EDTA and then freeze-dried. Sperm samples were stored at 4°C as room temperature, and the analysis of DNA damage was performed after a month and 5 months of storage using a Sperm Chromatin Dispersion test. We found four different sperm populations according to the size of the halos around the sperm head: (1) absent halo, (2) <6 μm, (3) 6-10 μm, (4) >10 μm. All of them coexisted in each freeze-dried dog semen samples and differed significantly among different treatments. The highest percentage of spermatozoa with halo >10 μm was obtained when the semen samples were freeze-dried in EDTA medium and stored at room temperature for five months. Results suggested that both, the kind of chelating agent as well as storage temperature and period, influenced DNA integrity of freeze-dried dog sperm. Copyright © 2015 Elsevier Inc. All rights reserved.

Redistribution of ice nuclei between cloud and rain droplets: Parameterization and application to deep convective clouds

NASA Astrophysics Data System (ADS)

Paukert, M.; Hoose, C.; Simmel, M.

2017-03-01

In model studies of aerosol-dependent immersion freezing in clouds, a common assumption is that each ice nucleating aerosol particle corresponds to exactly one cloud droplet. In contrast, the immersion freezing of larger drops—"rain"—is usually represented by a liquid volume-dependent approach, making the parameterizations of rain freezing independent of specific aerosol types and concentrations. This may lead to inconsistencies when aerosol effects on clouds and precipitation shall be investigated, since raindrops consist of the cloud droplets—and corresponding aerosol particles—that have been involved in drop-drop-collisions. Here we introduce an extension to a two-moment microphysical scheme in order to account explicitly for particle accumulation in raindrops by tracking the rates of selfcollection, autoconversion, and accretion. This provides a direct link between ice nuclei and the primary formation of large precipitating ice particles. A new parameterization scheme of drop freezing is presented to consider multiple ice nuclei within one drop and effective drop cooling rates. In our test cases of deep convective clouds, we find that at altitudes which are most relevant for immersion freezing, the majority of potential ice nuclei have been converted from cloud droplets into raindrops. Compared to the standard treatment of freezing in our model, the less efficient mineral dust-based freezing results in higher rainwater contents in the convective core, affecting both rain and hail precipitation. The aerosol-dependent treatment of rain freezing can reverse the signs of simulated precipitation sensitivities to ice nuclei perturbations.

Microbial analysis and survey test of gamma-irradiated freeze-dried fruits for patient's food

NASA Astrophysics Data System (ADS)

Park, Jae-Nam; Sung, Nak-Yun; Byun, Eui-Hong; Byun, Eui-Baek; Song, Beom-Seok; Kim, Jae-Hun; Lee, Kyung-A.; Son, Eun-Joo; Lyu, Eun-Soon

2015-06-01

This study examined the microbiological and organoleptic qualities of gamma-irradiated freeze-dried apples, pears, strawberries, pineapples, and grapes, and evaluated the organoleptic acceptability of the sterilized freeze-dried fruits for hospitalized patients. The freeze-dried fruits were gamma-irradiated at 0, 1, 2, 3, 4, 5, 10, 12, and 15 kGy, and their quality was evaluated. Microorganisms were not detected in apples after 1 kGy, in strawberries and pears after 4 kGy, in pineapples after 5 kGy, and in grapes after 12 kGy of gamma irradiation. The overall acceptance score, of the irradiated freeze-dried fruits on a 7-point scale at the sterilization doses was 5.5, 4.2, 4.0, 4.1, and 5.1 points for apples, strawberries, pears, pineapples, and grapes, respectively. The sensory survey of the hospitalized cancer patients (N=102) resulted in scores of 3.8, 3.7, 3.9, 3.9, and 3.7 on a 5-point scale for the gamma-irradiated freeze-dried apples, strawberries, pears, pineapples, and grapes, respectively. The results suggest that freeze-dried fruits can be sterilized with a dose of 5 kGy, except for grapes, which require a dose of 12 kGy, and that the organoleptic quality of the fruits is acceptable to immuno-compromised patients. However, to clarify the microbiological quality and safety of freeze-dried fruits should be verified by plating for both aerobic and anaerobic microorganisms.

Effect of Freezing Rate and Microwave Thawing on Texture and Microstructural Properties of Potato (Solanum tuberosum).

PubMed

Phinney, David M; Frelka, John C; Wickramasinghe, Anita; Heldman, Dennis R

2017-04-01

Food freezing is a preservation process that works by lowering temperature while simultaneously decreasing water activity. It is accepted that although freezing preserves foods, it generally has a negative effect on textural quality. This research investigated the texture response of potatoes (Solanum tuberosum) as a function of time to freeze (defined as the time for the center temperature to reach -20 °C) and thawing process. Potatoes slices (6 mm) were blanched then frozen in an ethanol/carbon dioxide bath, a pilot scale high velocity air freezer (HVAF) and a still air freezer to achieve various times to freeze. Slices were stabilized at -20 °C and thawed by 2 methods; room temperature air and microwave. Afterwards, samples were allowed to come to room temperature prior to texture profile analysis (TPA). Results indicate a maximum texture loss of the potato was reached at a time to freeze of approximately 8 min (corresponding to the HVAF). The texture difference between room temperature and microwave thawing methods was not shown to be significant (P = 0.05). SEM images showed the cellular structure of the potato in a HVAF to be similar to that of the still air freezer, validating that the matrix was maximally damaged in both conditions. This work created a continuous quality loss model for the potato as a function of time to freeze and showed no textural benefit to high velocity over still air freezing. © 2017 Institute of Food Technologists®.

Handwriting Impairments in People With Parkinson's Disease and Freezing of Gait.

PubMed

Heremans, Elke; Nackaerts, Evelien; Broeder, Sanne; Vervoort, Griet; Swinnen, Stephan P; Nieuwboer, Alice

2016-11-01

Recent studies show that patients with Parkinson's disease (PD) and freezing of gait (FOG) experience motor problems outside their gait freezing episodes. Because handwriting is also a sequential movement, it may be affected in PD patients with FOG relative to those without. The current study aimed to assess the quality of writing in PD patients with and without FOG in comparison to healthy controls (CTs) during various writing tasks. Handwriting was assessed by the writing of cursive loops on a touch-sensitive writing tablet and by means of the Systematic Screening of Handwriting Difficulties (SOS) test in 30 PD patients with and without freezing and 15 healthy age-matched CTs. The tablet tests were performed at 2 different sizes, either continuously or alternatingly, as indicated by visual target lines. Patients with freezing showed decreased writing amplitudes and increased variability compared with CTs and patients without freezing on the writing tablet tests. Writing problems were present during both tests but were more pronounced during writing at alternating compared with writing at continuous size. Patients with freezing also had a higher total score on the SOS test than patients without freezing and CTs, reflecting more extensive handwriting problems, particularly with writing fluency. Writing is more severely affected in PD patients with FOG than in those without FOG. These results indicate that deficient movement sequencing and adaptation is a generic problem in patients with FOG. © The Author(s) 2016.

Fuel freeze-point investigations. Final report, September 1982-March 1984

DOE Office of Scientific and Technical Information (OSTI.GOV)

Desmarais, L.A.; Tolle, F.F.

1984-07-01

The objective of this program was to conduct a detailed assessment of the low-temperature environment to which USAF aircraft are exposed for the purpose of defining a maximum acceptable fuel freeze-point and also to define any operational changes required with the use of a high freeze-point fuel. A previous study of B-52, C-141, and KC-135 operational missions indicated that the -58 C freeze point specification was too conservative. Based on recommendations resulting from the previous program, several improvements in the method of analysis were made, such as: expansion of the atmospheric temperature data base, the addition of ground temperature analysis,more » the addition of fuel-freezing analysis to the one-dimensional fuel-temperature computer program, and the examination of heat transfer in external fuel tanks, such as pylon or tip tanks. The B-52, C-141, and KC-135 mission were analyzed again, along with the operational missions of two tactical airplanes, the A-10 and F-15; -50C was determined to be the maximum allowable freeze point for a general-purpose USAF aviation turbine fuel. Higher freeze points can be tolerated if the probability of operational interference is acceptably low or if operational changes can be made. Study of atmospheric temperatures encountered for the missions of the five-study aircraft indicates that a maximum freeze point of -48 C would not likely create any operational difficulties in Northern Europe.« less

Freeze-thaw and high-voltage discharge allow macromolecule uptake into ileal brush-border vesicles

DOE Office of Scientific and Technical Information (OSTI.GOV)

Donowitz, M.; Emmer, E.; McCullen, J.

1987-06-01

High-voltage discharge or one cycle of freeze-thawing are shown to transiently permeabilize rabbit ileal brush-border membrane vesicles to macromolecules. Uptake of the radiolabeled macromolecule dextran, mol wt 70,000, used as a marker for vesicle permeability, was determined by a rapid filtration technique, with uptake defined as substrate associated with the vesicle and releasable after incubation of vesicles with 0.1% saponin. Dextran added immediately after electric shock (2000 V) or at the beginning of one cycle of freeze-thawing was taken up approximately eightfold compared with control. ATP also was taken up into freeze-thawed vesicles, whereas there was no significant uptake intomore » control vesicles. The increase in vesicle permeability was reversible, based on Na-dependent D-glucose uptake being decreased when studied 5 but not 15 min after electric shock, and was not significantly decreased after completion of one cycle of freeze-thawing. In addition, adenosine 3',5'-cyclic monophosphate and Ca/sup 2 +/-calmodulin-dependent protein kinase activity were similar in control vesicles and vesicles exposed to high-voltage discharge or freeze-thawing. Also, vesicles freeze-thawed with (/sup 32/P)ATP demonstrated increased phosphorylation compared with nonfrozen vesicles, while freeze-thawing did not alter vesicle protein as judged by Coomassie blue staining. These techniques should allow intestinal membrane vesicles to be used for studies of intracellular control of transport processes, for instance, studies of protein kinase regulation of transport.« less

Physicochemical interaction mechanism between nanoparticles and tetrasaccharides (stachyose) during freeze-drying.

PubMed

Kamiya, Seitaro; Nakashima, Kenichiro

2017-12-01

Nanoparticle suspensions are thermodynamically unstable and subject to aggregation. Freeze-drying on addition of saccharides is a useful method for preventing aggregation. In the present study, tetrasaccharides (stachyose) was employed as an additive. In addition, we hypothesize the interactive mechanism between stachyose and the nanoparticles during freeze-drying for the first time. The mean particle size of the rehydrated freeze-dried stachyose-containing nanoparticles (104.7 nm) was similar to the initial particle size before freeze-drying (76.8 nm), indicating that the particle size had been maintained. The mean particle size of the rehydrated normal-dried stachyose-containing nanoparticles was 222.2 nm. The powder X-ray diffraction of the freeze-dried stachyose-containing nanoparticles revealed a halo pattern. The powder X-ray diffraction of the normally dried stachyose-containing nanoparticles produced mainly a halo pattern and a partial peak. These results suggest an interaction between the nanoparticles and stachyose, and that this relationship depends on whether the mixture is freeze-dried or dried normally. In the case of normal drying, although most molecules cannot move rapidly thereby settling irregularly, some stachyose molecules can arrange regularly leading to some degree of crystallization and potentially some aggregation. In contrast, during freeze-drying, the moisture sublimed, while the stachyose molecules and nanoparticles were immobilized in the ice. After sublimation, stachyose remained in the space occupied by water and played the role of a buffer material, thus preventing aggregation.

Influence of Rapid Freeze-Thaw Cycling on the Mechanical Properties of Sustainable Strain-Hardening Cement Composite (2SHCC)

PubMed Central

Jang, Seok-Joon; Rokugo, Keitetsu; Park, Wan-Shin; Yun, Hyun-Do

2014-01-01

This paper provides experimental results to investigate the mechanical properties of sustainable strain-hardening cement composite (2SHCC) for infrastructures after freeze-thaw actions. To improve the sustainability of SHCC materials in this study, high energy-consumptive components—silica sand, cement, and polyvinyl alcohol (PVA) fibers—in the conventional SHCC materials are partially replaced with recycled materials such as recycled sand, fly ash, and polyethylene terephthalate (PET) fibers, respectively. To investigate the mechanical properties of green SHCC that contains recycled materials, the cement, PVA fiber and silica sand were replaced with 10% fly ash, 25% PET fiber, and 10% recycled aggregate based on preliminary experimental results for the development of 2SHCC material, respectively. The dynamic modulus of elasticity and weight for 2SHCC material were measured at every 30 cycles of freeze-thaw. The effects of freeze-thaw cycles on the mechanical properties of sustainable SHCC are evaluated by conducting compressive tests, four-point flexural tests, direct tensile tests and prism splitting tests after 90, 180, and 300 cycles of rapid freeze-thaw. Freeze-thaw testing was conducted according to ASTM C 666 Procedure A. Test results show that after 300 cycles of freezing and thawing actions, the dynamic modulus of elasticity and mass loss of damaged 2SHCC were similar to those of virgin 2SHCC, while the freeze-thaw cycles influence mechanical properties of the 2SHCC material except for compressive behavior. PMID:28788522

Migration of Water in Litopenaeus Vannamei Muscle Following Freezing and Thawing.

PubMed

Deng, Qi; Wang, Yaling; Sun, Lijun; Li, Jianrong; Fang, Zhijia; Gooneratne, Ravi

2018-06-15

Water and protein are major constituents of shrimp, any changes in protein and the state of water influence the quality of shrimp. Therefore, a study to examine the law of moisture migration and protein denaturation under different freezing and thawing conditions is important. The proton density images of thawed frozen-shrimp revealed that the water loss during quick-freezing was much greater than that during slow freezing or microfreezing. At room temperature (25 °C), the water loss from brine-thawing was more than still-water thawing and still-water thawing was more than thawing spontaneously. Freezing-thawing resulted in uniform water redistribution in shrimp muscle. Nuclear magnetic resonance technology (low field magnetic imaging) was used to directly monitor the dynamic processes of fluidity state in shrimp and indirectly monitor protein denaturation and thereby determine the optimal method of freezing-thawing shrimp. Our research showed that microfreezing preservation minimized weight loss, juice leakage and protein denaturation in shrimp muscle during thawing. Water is one of the major components in most organs and is an important factor that influences the shrimp muscle quality. Water migration patterns and subsequent effects on the shrimp muscle under different freezing and thawing conditions were examined using low field nuclear magnetic resonance (NMR) technology. This research provides a theoretical foundation for shrimp processing plants to improve the freezing and thawing process to obtain optimal quality and flavor of shrimp products. © 2018 Institute of Food Technologists®.

Protective effect of sugars on storage stability of microwave freeze-dried and freeze-dried Lactobacillus paracasei F19.

PubMed

Ambros, S; Hofer, F; Kulozik, U

2018-05-31

Microwave freeze drying in comparison to conventional freeze drying allows for intensification of the preservation process of lactic acid bacteria without imposing additional processing stress. Viability as a function of storage time of microwave freeze-dried Lactobacillus paracasei ssp. paracasei F19 was investigated in comparison to conventionally lyophilized bacteria of the same strain. Further, the impact of the protectants, sorbitol, trehalose and maltodextrin, on shelf life was analyzed. The highest inactivation rates of 0.035 and 0.045 d -1 , respectively, were found for cultures without protectants. Thus, all additives were found to exhibit a protective effect during storage with inactivation rates between 0.015 and 0.040 d -1 . Although trehalose and maltodextrin samples were in the glassy state during storage, in contrast to samples containing sorbitol as protectant, the best protective effect could be found for sorbitol with the lowest inactivation rate of 0.015 d -1 . Due to its low molecular weight, it might protect cells owing to better adsorption to the cytoplasma membrane. Sorbitol additionally shows antioxidative properties. Storage behavior of microwave freeze-dried cultures follows the typical behavior of a product dried by conventional lyophilization. No significant influence of the drying technique on storage behavior was detected. General findings concerning storage behavior in freeze drying are likely to be applicable in microwave freeze drying with only slight adjustments. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

3 CFR - Pay Freeze

Code of Federal Regulations, 2010 CFR

2010-01-01

... 3 The President 1 2010-01-01 2010-01-01 false Pay Freeze Presidential Documents Other Presidential Documents Memorandum of January 21, 2009 Pay Freeze Memorandum for the Assistant to the President and Chief... the White House staff forgo pay increases until further notice. Accordingly, as a signal of our shared...

Stress, deformation and micromorphological aspects of soil freezing under laboratory conditions

NASA Astrophysics Data System (ADS)

Jetchick, Elizabeth

In this thesis, frost heave is viewed as a process resulting from the interactions between thermodynamic conditions, soil environment controls such as texture, stress/deformation conditions and soil microstructure. A series of laboratory experiments was devised to investigate the links between these aspects. Because a limited number of studies exist on the development of internal stresses and strains in freezing soil, the work focussed on obtaining rheological data using conventional soil strain gauges and prototype stress transducers. A fine-grained unstructured silt was placed in a column (30 cm diameter by 100 cm length) and subjected to freezing and freeze-thaw cycles from the top down, lasting up to three months. Heat and water flows, as well as stresses and strains were monitored. The frozen soil was sectioned at the end of four of the experiments to examine the soil fabrics that had developed. From the experimental results, schematic stress and strain curves are proposed. For a single freeze cycle, compressive normal and tensile normal stresses were recorded simultaneously by the measuring devices within the freezing soil profile. Ice lens inception took place when the stress field changed, a condition which occurred either at the frost front level or at the base of the growing ice lens. Negative and positive strains reflected the different stress states that were sustained below and above the freezing front. Negative strains or soil consolidation took place as stresses increased before the passage of the frost line. Negligible soil strains were recorded as maximum soil consolidation was attained, before soil expansion. Distinct positive strain patterns indicating secondary and continuing heave, were recorded simultaneously throughout a thickness of soil, over a range of temperatures. Ice lens growth mostly took place as secondary frost heave, but continuing heave was measured, and the temperature conditions for both types of heave were determined. During subsequent freeze-thaw cycles, the stress patterns upon freezing were more complex in the second and third cycles due to previous soil structuration. At thaw, the stress pattern was uniform although positive strains in excess of those generated at freezing were recorded over the course of a few hours. Specific soil fabrics and features were evident from a single freeze cycle and for freeze-thaw conditions. Formation mechanisms are proposed for certain fabrics and features. A zonation with depth of these fabrics can be linked to the stress strain history of the soil, revealing the links and feedbacks between rheological processes and cryogenic soil structures.

Site-directed Nanotherapeutics to Abrogate RRMS and Promote Remyelination Repair (Rev)

DTIC Science & Technology

2013-03-01

in the labs additionally delayed the onset of the practical work.  Long lasting defect in freeze / dryer prevented a more in depth investigation of...drug content and reduced functionality of the peptides. Freezing of the NP without cryoprotection leads to disassembly; freezing with a cryoprotectant

Investigation of the impact of nanotechnology on the freeze-thaw durability of concrete containing d-cracking aggregates : [technical summary].

DOT National Transportation Integrated Search

2015-05-01

Freezing and thawing damage is the most common cause of distress in : Kansas pavements. Many locally available aggregates in Kansas do not : meet current standards for use in concrete pavements because of poor : freeze-thaw durability. The use of nan...

Investigation of the impact of nanotechnology on the freeze-thaw durability of concrete containing d-cracking aggregates.

DOT National Transportation Integrated Search

2015-05-01

Freezing and thawing damage is the most common cause of distress in Kansas pavements. Many : locally available aggregates in Kansas do not meet current standards for use in concrete pavements because : of poor freeze-thaw durability. The use of nanot...

7 CFR 58.638 - Freezing the mix.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 3 2013-01-01 2013-01-01 false Freezing the mix. 58.638 Section 58.638 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... Procedures § 58.638 Freezing the mix. After the mix enters the freezer, it shall be frozen as rapidly as...