Search results for: thawing
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 51

Search results for: thawing

21 Improving Concrete Properties with Fibers Addition

Authors: E. Mello, C. Ribellato, E. Mohamedelhassan

Abstract:

This study investigated the improvement in concrete properties with addition of cellulose, steel, carbon and PET fibers. Each fiber was added at four percentages to the fresh concrete, which was moist-cured for 28-days and then tested for compressive, flexural and tensile strengths. Changes in strength and increases in cost were analyzed. Results showed that addition of cellulose caused a decrease between 9.8% and 16.4% in compressive strength. This range may be acceptable as cellulose fibers can significantly increase the concrete resistance to fire, and freezing and thawing cycles. Addition of steel fibers to concrete increased the compressive strength by up to 20%. Increases 121.5% and 80.7% were reported in tensile and flexural strengths respectively. Carbon fibers increased flexural and tensile strengths by up to 11% and 45%, respectively. Concrete strength properties decreased after the addition of PET fibers. Results showed that improvement in strength after addition of steel and carbon fibers may justify the extra cost of fibers.

Keywords: concrete, compressive strength, fibers, flexural strength, tensile strength

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20 Prediction of Permeability of Frozen Unsaturated Soil Using Van Genuchten Model and Fredlund-Xing Model in Soil Vision

Authors: Bhavita S. Dave, Jaimin Vaidya, Chandresh H. Solanki, Atul K.

Abstract:

To measure the permeability of a soil specimen, one of the basic assumptions of Darcy's law is that the soil sample should be saturated. Unlike saturated soils, the permeability of unsaturated soils cannot be found using conventional methods as it does not follow Darcy's law. Many empirical models, such as the Van Genuchten Model and Fredlund-Xing Model were suggested to predict permeability value for unsaturated soil. Such models use data from the soil-freezing characteristic curve to find fitting parameters for frozen unsaturated soils. In this study, soil specimens were subjected to 0, 1, 3, and 5 freezing-thawing (F-T) cycles for different degrees of saturation to have a wide range of suction, and its soil freezing characteristic curves were formulated for all F-T cycles. Changes in fitting parameters and relative permeability with subsequent F-T cycles are presented in this paper for both models.

Keywords: frozen unsaturated soil, Fredlund Xing model, soil-freezing characteristic curve, Van Genuchten model

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19 Impact of Propolis on Cryopreservation of Arctic Charr (Salvelinus alpinus) Sperm

Authors: K. A. El-Battawy, E. Brannas

Abstract:

Cryopreservation of sperm causes damages and adversely affected sperm motility and viability resulting in lower hatching rates. The aim of this study is to determine whether propolis has potential protective effect on cryopreservation and fertilization ability of spermatozoa of Salvelinusalpinus. The extenders were prepared by using simple glucose solution (0.3 M glucose) to which 10% Me2SO added with different levels of propolis (0.4, 0.8 and 1 mg/ ml) and 10% egg yolk (as a control without propolis). The pooled semen samples diluted at the ratio of 1:3 by the extenders were subjected to cryopreservation. The percentage and duration of motility and fertilization tests of cryopreserved sperm samples have been done immediately after thawing and compared with control and fresh semen. The extenders containing propolis showed higher percentage motility and motility duration than control group (P < 0.05). Especially the group II (0.8 mg/ ml propolis) and the group III (1 mg/ ml propolis) showed significant positive effects on both post thaw motility and hatching ability. In conclusion, this study confirms that the propolis is an appropriate cryoptrotective agent in fish semen and it maintained the integrity of the spermatozoa during the cryopreservation process.

Keywords: propolis, arctic charr, semen, cryopreservation

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18 Application of Artificial Ground-Freezing to Construct a Passenger Interchange Tunnel for the Subway Line 14 in Paris, France

Authors: G. Lancellotta, G. Di Salvo, A. Rigazio, A. Davout, V. Pastore, G. Tonoli, A. Martin, P. Jullien, R. Jagow-Klaff, R. Wernecke

Abstract:

Artificial ground freezing (AGF) technique is a well-proven soil improvement approach used worldwide to construct shafts, tunnels and many other civil structures in difficult subsoil or ambient conditions. As part of the extension of Line 14 of the Paris subway, a passenger interchange tunnel between the new station at Porte de CI ichy and the new Tribunal the Grand Instance has been successfully constructed using this technique. The paper presents the successful application of AGF by Liquid Nitrogen and Brine implemented to provide structural stability and groundwater cut-off around the passenger interchange tunnel. The working conditions were considered to be rather challenging, due to the proximity of a hundred-year-old existing service tunnel of the Line 13, and subsoil conditions on site. Laboratory tests were carried out to determine the relevant soil parameters for hydro-thermal-mechanical aspects and to implement numerical analyses. Monitoring data were used in order to check and control the development and the efficiency of the freezing process as well as to back analyze the parameters assumed for the design, both during the freezing and thawing phases.

Keywords: artificial ground freezing, brine method, case history, liquid nitrogen

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17 Improvement of Mechanical Properties of Saline Soils by Fly Ash: Effect of Freeze-Thaw Cycles

Authors: Zhuo Cheng, Gaohang Cui, Yang Zheng, Zhiqiang-Pan

Abstract:

To explore the effect of freeze-thaw cycles on saline soil mechanical properties of fly ash, this study examined the influence of different numbers of freezing and thawing cycles, fly ash content, and moisture content of saline soil in unconfined compression tests and triaxial shear tests. With increased fly ash content, the internal friction angle, cohesion, unconfined compressive strength, and shear strength of the improved soil increased at first and then decreased. Using the Desk-Expert 8.0 software and based on significance analysis theory, the number of freeze-thaw cycles, fly ash content, water content, and the interactions between various factors on the mechanical properties of saline soil were studied. The results showed that the number of freeze-thaw cycles had a significant effect on the mechanical properties of saline soil, while the fly ash content had a weakly significant effect. At the same time, interaction between the number of freeze-thaw cycles and the water content had a significant effect on the unconfined compressive strength and the cohesion of saline soil, and the interaction between fly ash content and the number of freeze-thaw cycles only had a significant effect on the unconfined compressive strength.

Keywords: fly ash, saline soil, seasonally frozen area, significance analysis, qualitative analysis

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16 Durability of a Cementitious Matrix Based on Treated Sediments

Authors: Mahfoud Benzerzour, Mouhamadou Amar, Amine Safhi, Nor-Edine Abriak

Abstract:

Significant volumes of sediment are annually dredged in France and all over the world. These materials may, in fact, be used beneficially as supplementary cementitious material. This paper studies the durability of a new cement matrix based on marine dredged sediment of Dunkirk-Harbor (north of France). Several techniques are used to characterize the raw sediment such as physical properties, chemical analyses, and mineralogy. The XRD analysis revealed quartz, calcite, kaolinite as main mineral phases. In order to eliminate organic matter and activate some of those minerals, the sediment is calcined at a temperature of 850°C for 1h. Moreover, four blended mortars were formulated by mixing a portland cement (CEM I 52,5 N) and the calcined sediment as partial cement substitute (0%, 10%, 20% and 30%). Reference mortars, based on the blended cement, were then prepared. This re-use cannot be substantiating and efficient without a durability study. In this purpose, the following tests, mercury porosity, accessible water porosity, chloride permeability, freezing and thawing, external sulfate attack, alkali aggregates reaction, compressive and bending strength tests were conducted on those mortars. The results of most of those tests evidenced the fact that the mortar that contains 10% of the treated sediment is efficient and durable as the reference mortar itself. That would infer that the presence of these calcined sediment improves mortar general behavior.

Keywords: sediment, characterization, calcination, substitution, durability

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15 Influence of Ground Granulated Blast Furnace Slag on Geotechnical Characteristics of Jarosite Waste

Authors: Chayan Gupta, Arun Prasad

Abstract:

The quick evolution of industrialization causes the scarcity of precious land. Thus, it is vital need to influence the R&D societies to achieve sustainable, economic and social benefits from huge utilization of waste for universal aids. The current study promotes the influence of steel industries waste i.e. ground granulated blast furnace slag (GGBS) in geotechnical properties of jarosite waste (solid waste residues produced from hydrometallurgy operations involved in extraction of Zinc). Numerous strengths tests (unconfined compression (qu) and splitting tensile strength (qt)) are conducted on jarosite-GGBS blends (GGBS, 10-30%) with different curing periods (7, 28 & 90 days). The results indicate that both qu and qt increase with the increase in GGBS content along with curing periods. The increased strength with the addition of GGBS is also observed from microstructural study, which illustrates the occurrence of larger agglomeration of jarosite-GGBS blend particles. The Freezing-Thawing (F-T) durability analysis is also conducted for all the jarosite-GGBS blends and found that the reduction in unconfined compressive strength after five successive F-T cycles enhanced from 62% (natural jarosite) to 48, 42 and 34% at 7, 14 and 28 days curing periods respectively for stabilized jarosite-GGBS samples containing 30% GGBS content. It can be concluded from this study that blending of cementing additives (GGBS) with jarosite waste resulted in a significant improvement in geotechnical characteristics.

Keywords: jarosite, GGBS, strength characteristics, microstructural study, durability analysis

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14 The Effect of Curcumin on Cryopreserved Bovine Semen

Authors: Eva Tvrdá, Marek Halenár, Hana Greifová, Alica Mackovich, Faridullah Hashim, Norbert Lukáč

Abstract:

Oxidative stress associated with semen cryopreservation may result in lipid peroxidation (LPO), DNA damage and apoptosis, leading to decreased sperm motility and fertilization ability. Curcumin (CUR), a natural phenol isolated from Curcuma longa Linn. has been presented as a possible supplement for a more effective semen cryopreservation because of its antioxidant properties. This study focused to evaluate the effects of CUR on selected oxidative stress parameters in cryopreserved bovine semen. 20 bovine ejaculates were split into two aliquots and diluted with a commercial semen extender containing CUR (50 μmol/L) or no supplement (control), cooled to 4 °C, frozen and kept in liquid nitrogen. Frozen straws were thawed in a water bath for subsequent experiments. Computer assisted semen analysis was used to evaluate spermatozoa motility, and reactive oxygen species (ROS) generation was quantified by using luminometry. Superoxide generation was evaluated with the NBT test, and LPO was assessed via the TBARS assay. CUR supplementation significantly (P<0.001) increased the spermatozoa motility and provided a significantly higher protection against ROS (P<0.001) or superoxide (P<0.01) overgeneration caused by semen freezing and thawing. Furthermore, CUR administration resulted in a significantly (P<0.01) lower LPO of the experimental semen samples. In conclusion, CUR exhibits significant ROS-scavenging activities which may prevent oxidative insults to cryopreserved spermatozoa and thus may enhance the post-thaw functional activity of male gametes.

Keywords: bulls, cryopreservation, curcumin, lipid peroxidation, reactive oxygen species, spermatozoa

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13 The Methanotrophic Activity in a Landfill Bio-Cover through a Subzero Winter

Authors: Parvin Berenjkar, Qiuyan Yuan, Richard Sparling, Stan Lozecznik

Abstract:

Landfills highly contribute to anthropological global warming through CH₄ emissions. Landfills are usually capped by a conventional soil cover to control the migration of gases. Methane is consumed by CH₄-oxidizing microorganisms known as methanotrophs that naturally exist in the landfill soil cover. The growth of methanotrophs can be optimized in a bio-cover that typically consists of a gas distribution layer (GDL) to homogenize landfill gas fluxes and an overlying oxidation layer composed of suitable materials that support methanotrophic populations. Materials such as mature yard waste composts can provide an inexpensive and favourable porous support for the growth and activity of methanotrophs. In areas with seasonal cold climates, it is valuable to know if methanotrophs in a bio-cover can survive in winter until the next spring, and how deep they are active in the bio-cover to mitigate CH₄. In this study, a pilot bio-cover was constructed in a closed landfill cell in Winnipeg that has a very cold climate in Canada. The bio-cover has a surface area of 2.5 m x 3.5 m and 1.5 m of depth, filled with 50 cm of gravel as a GDL and 70 cm of biosolids compost amended with yard and leaf waste compost. The observed in situ potential of methanotrophs for CH₄ oxidation was investigated at a specific period of time from December 2016 to April 2017 as well as November 2017 to April 2018, when the transition to surface frost and thawing happens in the bio-cover. Compost samples taken from different depths of the bio-cover were incubated in the laboratory under standardized conditions; an optimal air: methane atmosphere, at 22ºC, but at in situ moisture content. Results showed that the methanotrophs were alive oxidizing methane without a lag, indicating that there was the potential for methanotrophic activity at some depths of the bio-cover.

Keywords: bio-cover, global warming, landfill, methanotrophic activity

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12 Old Swimmers Tire Quickly: The Effect of Time on Quality of Thawed versus Washed Sperm

Authors: Emily Hamilton, Adiel Kahana, Ron Hauser, Shimi Barda

Abstract:

BACKGROUND: In the male fertility and sperm bank unit of Tel Aviv Sourasky medical center, women are treated with intrauterine insemination (IUI) using washed sperm from their partner or thawed sperm from a selected donor. In most cases, the women perform the IUI treatment in Sourasky, but sometimes they ask to undergo the insemination procedure in another clinic with their own fertility doctor. In these cases, the sperm sample is prepared at the Sourasky lab and the patient is inseminated after arriving to her doctor. Our laboratory has previously found that time negatively affects several parameters of thawed sperm, and we estimate that it has more severe and significant effect than on washed sperm. AIM: To examine the effect of time on the quality of washed sperm versus thawed sperm. METHODS: Sperm samples were collected from men referred for semen analysis. Each ejaculate was allowed to liquefy for at least 20 min at 37°C and analyzed for sperm motility and vitality percentage and DNA fragmentation index (Time 0). Subsequently, 1ml of the sample was divided into two parts, 1st part was washed only and the 2nd part was washed, frozen and thawed. Time 1 analysis occurred immediately after sperm washing or thawing. Time 2 analysis occurred 75 minutes after time 1. Statistical analysis was performed using Student t-test. P values<0.05 were considered significant. RESULTS: Preliminary data showed that time had a greater impact on the average percentages of sperm motility and vitality in thawed compared to washed sperm samples (26%±10% vs. 21%±10% and 21%±9% vs. 9%±10%, respectively). An additional trend towards increased average DNA fragmentation percentage in thawed samples compared to washed samples was observed (46%±18% vs. 25%±24%). CONCLUSION: Time negatively effects sperm quality. The effect is greater in thawed samples compared to fresh samples.

Keywords: ART, male fertility, sperm cryopreservation, sperm quality

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11 A Study of Growth Performance, Carcass Characteristic, Meat Quality and Association of Polymorphism in the ApoVLDL-II Gene with Fat Accumulation in the Female Broiler, Thai Native and Betong Chickens (KU Line)

Authors: C. Kridtayopas, W. Danvilai, P. Sopannarath, A. Kayan, W. Loongyai

Abstract:

Both Betong chicken (KU Line) and Thai Native chickens were the high quality of the meat and low carcass fat compared to broiler chickens. The objective of this study was to determine the growth performance, carcass characteristic, meat quality and association of polymorphism in the ApoVLDL-II gene with fat accumulation in the female broiler, Thai Native and Betong (KU line) chickens at 4-14 weeks. The chickens were used and reared under the same environment and management (100 chicks per breed). The results showed that body weight (BW) of broiler chickens was significantly higher than Thai Native and Betong (KU line) chickens (P < 0.01) through all the experiment. At 4-8 weeks of age, feed conversion ratio (FCR) of broiler chickens was significantly better than Thai Native and Betong (KU line) chickens (P < 0.01), then increased at week 8-14. The percentage of breast, abdominal fat and subcutaneous fat of broiler chickens was significantly greater than Thai Native and Betong (KU line) chickens (P < 0.01). However, Thai Native chickens showed the highest percentage of liver (P < 0.01) when compared to other breeds. In addition, the percentage of wing of Thai Native and Betong (KU line) chickens were significantly (P < 0.01) higher than broiler chickens. Meat quality was also determined and found that, pH of breast meat left from slaughter 45 minutes (pH45) and 24 hours (pH24) of broiler was significantly higher than Thai Native and Betong (KU line) (P < 0.01) whereas the percentage of drip loss, thawing loss, cooking loss and shear force was not significantly different between breeds. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to genotype the polymorphism in the ApoVLDL-II gene in the broiler, Thai Native and Betong (KU line) chickens. The results found that, the polymorphism in the ApoVLDL-II gene at VLDL6 loci was not associated with fat accumulation in those studied population.

Keywords: ApoVLDL-II gene, Betong (KU line) chickens, broiler chickens, carcass characteristic, growth performance, meat quality, Thai native chickens

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10 Liquid Nitrogen as Fracturing Method for Hot Dry Rocks in Kazakhstan

Authors: Sotirios Longinos, Anna Loskutova, Assel Tolegenova, Assem Imanzhussip, Lei Wang

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Hot, dry rock (HDR) has substantial potential as a thermal energy source. It has been exploited by hydraulic fracturing to extract heat and generate electricity, which is a well-developed technique known for creating the enhanced geothermal systems (EGS). These days, LN2 is being tested as an environmental friendly fracturing fluid to generate densely interconnected crevices to augment heat exchange efficiency and production. This study examines experimentally the efficacy of LN2 cryogenic fracturing for granite samples in Kazakhstan with immersion method. A comparison of two different experimental models is carried out. The first mode is rock heating along with liquid nitrogen treatment (heating with freezing time), and the second mode is multiple times of heating along with liquid nitrogen treatment (heating with LN2 freezing-thawing cycles). The experimental results indicated that with multiple heating and LN2-treatment cycles, the permeability of granite first ameliorates with increasing number of cycles and later reaches a plateau after a certain number of cycles. On the other hand, density, P-wave velocity, uniaxial compressive strength, elastic modulus, and tensile strength indicate a downward trend with increasing heating and treatment cycles. The thermal treatment cycles do not seem to have an obvious effect on the Poisson’s ratio. The changing rate of granite rock properties decreases as the number of cycles increases. The deterioration of granite primarily happens within the early few cycles. The heating temperature during the cycles shows an important influence on the deterioration of granite. More specifically, mechanical deterioration and permeability amelioration become more remarkable as the heating temperature increases.LN2 fracturing generates many positives compared to conventional fracturing methods such as little water consumption, requirement of zero chemical additives, lessening of reservoir damage, and so forth. Based on the experimental observations, LN2 can work as a promising waterless fracturing fluid to stimulate hot, dry rock reservoirs.

Keywords: granite, hydraulic fracturing, liquid nitrogen, Kazakhstan

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9 Everolimus Loaded Polyvinyl Alcohol Microspheres for Sustained Drug Delivery in the Treatment of Subependymal Giant Cell Astrocytoma

Authors: Lynn Louis, Bor Shin Chee, Marion McAfee, Michael Nugent

Abstract:

This article aims to develop a sustained release formulation of microspheres containing the mTOR inhibitor Everolimus (EVR) using Polyvinyl alcohol (PVA) to enhance the bioavailability of the drug and to overcome poor solubility characteristics of Everolimus. This paper builds on recent work in the manufacture of microspheres using the sessile droplet technique by freezing the polymer-drug solution by suspending the droplets into pre-cooled ethanol vials immersed in liquid nitrogen. The spheres were subjected to 6 freezing cycles and 3 freezing cycles with thawing to obtain proper geometry, prevent aggregation, and achieve physical cross-linking. The prepared microspheres were characterised for surface morphology by SEM, where a 3-D porous structure was observed. The in vitro release studies showed a 62.17% release over 12.5 days, indicating a sustained release due to good encapsulation. This result is comparatively much more than the 49.06% release achieved within 4 hours from the solvent cast Everolimus film as a control with no freeze-thaw cycles performed. The solvent cast films were made in this work for comparison. A prolonged release of Everolimus using a polymer-based drug delivery system is essential to reach optimal therapeutic concentrations in treating SEGA tumours without systemic exposure. These results suggest that the combination of PVA and Everolimus via a rheological synergism enhanced the bioavailability of the hydrophobic drug Everolimus. Physical-chemical characterisation using DSC and FTIR analysis showed compatibility of the drug with the polymer, and the stability of the drug was maintained owing to the high molecular weight of the PVA. The obtained results indicate that the developed PVA/EVR microsphere is highly suitable as a potential drug delivery system with improved bioavailability in treating Subependymal Giant cell astrocytoma (SEGA).

Keywords: drug delivery system, everolimus, freeze-thaw cycles, polyvinyl alcohol

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8 Cryopreservation of Ring-Necked Pheasant (Phasianus colchicus) Semen for Establishing Cryobank

Authors: Rida Pervaiz, Bushra Allah Rakha, Muhammad Sajjad Ansari, Shamim Akhter, Kainat Waseem, Sumiyyah Zuha, Tooba Javed

Abstract:

Ring-necked pheasant (Phasianus colchicus) belongs to order Galliformes and family Phasianidae. It has been recognized as the most hunted bird due to its attractive colorful appearance and meat. Loss of habitat and hunting pressure has caused population fluctuations in the native range. Under these circumstances, this species can be conserved by employing ex-situ in vitro conservation techniques. Captive breeding, in combination with semen cryobanking is the most appropriate option to conserve/propagate this species without deteriorating the genetic diversity. Cryopreservation protocols of adequate efficiency are necessary to establish semen cryobanking for a species. Therefore, present study was designed to devise an efficient extender for cryopreservation of ring-necked pheasant semen. For this purpose, a range of extenders (Beltsville Poultry, red fowl, Lake, EK, Tselutin Poultry and Chicken semen extenders) were evaluated for cryopreservation of ring-necked pheasant semen. Semen collected from 10 cocks, diluted in the Beltsville Poultry (BPSE), Red Fowl (RFE), Lake (LE), EK (EKE), Tselutin Poultry (TPE) and Chicken Semen (CSE) extenders and cryopreserved. Glycerol (10%) was added to semen at 4°C, equilibrated for 10 min, filled in 0.5 mL French straws, kept over liquid nitrogen vapors for 10 min, cryopreserved in LN2 and stored. Sperm motility (%), viability (%), live/dead ratio (%), plasma membrane (%) and DNA Integrity (%) were evaluated at post-dilution, post-cooling, post-equilibration and post-thawing stage of cryopreservation. Sperm motility (83.8 ± 3.1; 81.3 ± 3.8; 73.8 ± 2.4; 62.5 ± 1.4), viability (79.0 ± 1.7; 75.5 ± 1.6; 69.5 ± 2.3; 65.5 ± 2.4), live/dead ratio (80.5 ± 5.7; 77.3 ± 4.9; 76.0 ± 2.7; 68.3 ± 2.3), plasma membrane (74.5 ± 2.9; 73.8 ± 3.4; 71.3 ± 2.3; 75.0 ± 3.4) and DNA integrity (78.3 ± 1.7; 73.0 ± 1.2; 68.0 ± 2.0; 63.0 ± 2.5) at all four stages of cryopreservation were recorded higher (P < 0.05) in red fowl extender compared to all experimental extenders. It is concluded that red fowl extender is the best extender for cryopreservation of ring-necked pheasant semen and can be used in establishing cryobank for ex situ conservation.

Keywords: ring-necked pheasant; extenders; cryopreservation; semen quality; DNA integrity

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7 Properties of Adipose Tissue Derived Mesenchymal Stem Cells with Long-Term Cryopreservation

Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

Abstract:

Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: mesenchymal stem cells, cryopreservation, stemness, senescence

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6 Calcein Release from Liposomes Mediated by Phospholipase A₂ Activity: Effect of Cholesterol and Amphipathic Di and Tri Blocks Copolymers

Authors: Marco Soto-Arriaza, Eduardo Cena-Ahumada, Jaime Melendez-Rojel

Abstract:

Background: Liposomes have been widely used as a model of lipid bilayer to study the physicochemical properties of biological membrane, encapsulation, transport and release of different molecules. Furthermore, extensive research has focused on improving the efficiency in the transport of drugs, developing tools that improve the release of the encapsulated drug from liposomes. In this context, the enzymatic activity of PLA₂, despite having been shown to be an effective tool to promote the release of drugs from liposomes, is still an open field of research. Aim: The aim of the present study is to explore the effect of cholesterol (Cho) and amphipathic di- and tri-block copolymers, on calcein release mediated by enzymatic activity of PLA2 in Dipalmitoylphosphatidylcholine (DPPC) liposomes under physiological conditions. Methods: Different dispersions of DPPC, cholesterol, di-block POE₄₅-PCL₅₂ or tri-block PCL₁₂-POE₄₅-PCL₁₂ were prepared by the extrusion method after five freezing/thawing cycles; in Phosphate buffer 10mM pH 7.4 in presence of calcein. DPPC liposomes/Calcein were centrifuged at 15000rpm 10 min to separate free calcein. Enzymatic activity assays of PLA₂ were performed at 37°C using the TBS buffer pH 7.4. The size distribution, polydispersity, Z-potential and Calcein encapsulation of DPPC liposomes was monitored. Results: PLA₂ activity showed a slower kinetic of calcein release up to 20 mol% of cholesterol, evidencing a minimum at 10 mol% and then a maximum at 18 mol%. Regardless of the percentage of cholesterol, up to 18 mol% a one-hundred percentage release of calcein was observed. At higher cholesterol concentrations, PLA₂ showed to be inefficient or not to be involved in calcein release. In assays where copolymers were added in a concentration lower than their cmc, a similar behavior to those showed in the presence of Cho was observed, that is a slower kinetic in calcein release. In both experimental approaches, a one-hundred percentage of calcein release was observed. PLA₂ was shown to be sensitive to the 4-(4-Octadecylphenyl)-4-oxobutenoic acid inhibitor and calcium, reducing the release of calcein to 0%. Cell viability of HeLa cells decreased 7% in the presence of DPPC liposomes after 3 hours of incubation and 17% and 23% at 5 and 15 hours, respectively. Conclusion: Calcein release from DPPC liposomes, mediated by PLA₂ activity, depends on the percentage of cholesterol and the presence of copolymers. Both, cholesterol up to 20 mol% and copolymers below it cmc could be applied to the regulation of the kinetics of antitumoral drugs release without inducing cell toxicity per se.

Keywords: amphipathic copolymers, calcein release, cholesterol, DPPC liposome, phospholipase A₂

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5 Analysis of the Properties of Hydrophobised Heat-Insulating Mortar with Perlite

Authors: Danuta Barnat-Hunek

Abstract:

The studies are devoted to assessing the effectiveness of hydrophobic and air entraining admixtures based on organ silicon compounds. Mortars with lightweight aggregate–perlite were the subjects of the investigation. The following laboratory tests were performed: density, open porosity, total porosity, absorptivity, capability to diffuse water vapour, compressive strength, flexural strength, frost resistance, sodium sulphate corrosion resistance and the thermal conductivity coefficient. The composition of the two mixtures of mortars was prepared: mortars without a hydrophobic admixture and mortars with cementitious waterproofing material. Surface hydrophobisation was produced on the mortars without a hydrophobic admixture using a methyl silicone resin, a water-based emulsion of methyl silicone resin in potassium hydroxide and alkyl-alkoxy-silane in organic solvents. The results of the effectiveness of hydrophobisation of mortars are the following: The highest absorption after 14 days of testing was shown by mortar without an agent (57.5%), while the lowest absorption was demonstrated by the mortar with methyl silicone resin (52.7%). After 14 days in water the hydrophobisation treatment of the samples proved to be ineffective. The hydrophobised mortars are characterized by an insignificant mass change due to freezing and thawing processes in the case of the methyl silicone resin – 1%, samples without hydrophobisation –5%. This agent efficiently protected the mortars against frost corrosion. The standard samples showed very good resistance to the pressure of sodium sulphate crystallization. Organosilicon compounds have a negative influence on the chemical resistance (weight loss about 7%). The mass loss of non-hydrophobic mortar was 2 times lower than mortar with the hydrophobic admixture. Hydrophobic and aeration admixtures significantly affect the thermal conductivity and the difference is mainly due to the difference in porosity of the compared materials. Hydrophobisation of the mortar mass slightly decreased the porosity of the mortar, and thus in an increase of 20% of its compressive strength. The admixture adversely affected the ability of the hydrophobic mortar – it achieved the opposite effect. As a result of hydrophobising the mass, the mortar samples decreased in density and had improved wettability. Poor protection of the mortar surface is probably due to the short time of saturating the sample in the preparation. The mortars were characterized by high porosity (65%) and water absorption (57.5%), so in order to achieve better efficiency, extending the time of hydrophobisation would be advisable. The highest efficiency was obtained for the surface hydrophobised with the methyl silicone resin.

Keywords: hydrophobisation, mortars, salt crystallization, frost resistance

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4 Peculiarities of Snow Cover in Belarus

Authors: Aleh Meshyk, Anastasiya Vouchak

Abstract:

On the average snow covers Belarus for 75 days in the south-west and 125 days in the north-east. During the cold season snowpack often destroys due to thaws, especially at the beginning and end of winter. Over 50% of thawing days have a positive mean daily temperature, which results in complete snow melting. For instance, in December 10% of thaws occur at 4 С mean daily temperature. Stable snowpack lying for over a month forms in the north-east in the first decade of December but in the south-west in the third decade of December. The cover disappears in March: in the north-east in the last decade but in the south-west in the first decade. This research takes into account that precipitation falling during a cold season could be not only liquid and solid but also a mixed type (about 10-15 % a year). Another important feature of snow cover is its density. In Belarus, the density of freshly fallen snow ranges from 0.08-0.12 g/cm³ in the north-east to 0.12-0.17 g/cm³ in the south-west. Over time, snow settles under its weight and after melting and refreezing. Averaged annual density of snow at the end of January is 0.23-0.28 g/сm³, in February – 0.25-0.30 g/сm³, in March – 0.29-0.36 g/сm³. Sometimes it can be over 0.50 g/сm³ if the snow melts too fast. The density of melting snow saturated with water can reach 0.80 g/сm³. Average maximum of snow depth is 15-33 cm: minimum is in Brest, maximum is in Lyntupy. Maximum registered snow depth ranges within 40-72 cm. The water content in snowpack, as well as its depth and density, reaches its maximum in the second half of February – beginning of March. Spatial distribution of the amount of liquid in snow corresponds to the trend described above, i.e. it increases in the direction from south-west to north-east and on the highlands. Average annual value of maximum water content in snow ranges from 35 mm in the south-west to 80-100 mm in the north-east. The water content in snow is over 80 mm on the central Belarusian highland. In certain years it exceeds 2-3 times the average annual values. Moderate water content in snow (80-95 mm) is characteristic of western highlands. Maximum water content in snow varies over the country from 107 mm (Brest) to 207 mm (Novogrudok). Maximum water content in snow varies significantly in time (in years), which is confirmed by high variation coefficient (Cv). Maximums (0.62-0.69) are in the south and south-west of Belarus. Minimums (0.42-0.46) are in central and north-eastern Belarus where snow cover is more stable. Since 1987 most gauge stations in Belarus have observed a trend to a decrease in water content in snow. It is confirmed by the research. The biggest snow cover forms on the highlands in central and north-eastern Belarus. Novogrudok, Minsk, Volkovysk, and Sventayny highlands are a natural orographic barrier which prevents snow-bringing air masses from penetrating inside the country. The research is based on data from gauge stations in Belarus registered from 1944 to 2014.

Keywords: density, depth, snow, water content in snow

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3 Early Melt Season Variability of Fast Ice Degradation Due to Small Arctic Riverine Heat Fluxes

Authors: Grace E. Santella, Shawn G. Gallaher, Joseph P. Smith

Abstract:

In order to determine the importance of small-system riverine heat flux on regional landfast sea ice breakup, our study explores the annual spring freshet of the Sagavanirktok River from 2014-2019. Seasonal heat cycling ultimately serves as the driving mechanism behind the freshet; however, as an emerging area of study, the extent to which inland thermodynamics influence coastal tundra geomorphology and connected landfast sea ice has not been extensively investigated in relation to small-scale Arctic river systems. The Sagavanirktok River is a small-to-midsized river system that flows south-to-north on the Alaskan North Slope from the Brooks mountain range to the Beaufort Sea at Prudhoe Bay. Seasonal warming in the spring rapidly melts snow and ice in a northwards progression from the Brooks Range and transitional tundra highlands towards the coast and when coupled with seasonal precipitation, results in a pulsed freshet that propagates through the Sagavanirktok River. The concentrated presence of newly exposed vegetation in the transitional tundra region due to spring melting results in higher absorption of solar radiation due to a lower albedo relative to snow-covered tundra and/or landfast sea ice. This results in spring flood runoff that advances over impermeable early-season permafrost soils with elevated temperatures relative to landfast sea ice and sub-ice flow. We examine the extent to which interannual temporal variability influences the onset and magnitude of river discharge by analyzing field measurements from the United States Geological Survey (USGS) river and meteorological observation sites. Rapid influx of heat to the Arctic Ocean via riverine systems results in a noticeable decay of landfast sea ice independent of ice breakup seaward of the shear zone. Utilizing MODIS imagery from NASA’s Terra satellite, interannual variability of river discharge is visualized, allowing for optical validation that the discharge flow is interacting with landfast sea ice. Thermal erosion experienced by sediment fast ice at the arrival of warm overflow preconditions the ice regime for rapid thawing. We investigate the extent to which interannual heat flux from the Sagavanirktok River’s freshet significantly influences the onset of local landfast sea ice breakup. The early-season warming of atmospheric temperatures is evidenced by the presence of storms which introduce liquid, rather than frozen, precipitation into the system. The resultant decreased albedo of the transitional tundra supports the positive relationship between early-season precipitation events, inland thermodynamic cycling, and degradation of landfast sea ice. Early removal of landfast sea ice increases coastal erosion in these regions and has implications for coastline geomorphology which stress industrial, ecological, and humanitarian infrastructure.

Keywords: Albedo, freshet, landfast sea ice, riverine heat flux, seasonal heat cycling

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2 Oncolytic Efficacy of Thymidine Kinase-Deleted Vaccinia Virus Strain Tiantan (oncoVV-TT) in Glioma

Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi

Abstract:

Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.

Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus

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1 Experimental Characterisation of Composite Panels for Railway Flooring

Authors: F. Pedro, S. Dias, A. Tadeu, J. António, Ó. López, A. Coelho

Abstract:

Railway transportation is considered the most economical and sustainable way to travel. However, future mobility brings important challenges to railway operators. The main target is to develop solutions that stimulate sustainable mobility. The research and innovation goals for this domain are efficient solutions, ensuring an increased level of safety and reliability, improved resource efficiency, high availability of the means (train), and satisfied passengers with the travel comfort level. These requirements are in line with the European Strategic Agenda for the 2020 rail sector, promoted by the European Rail Research Advisory Council (ERRAC). All these aspects involve redesigning current equipment and, in particular, the interior of the carriages. Recent studies have shown that two of the most important requirements for passengers are reasonable ticket prices and comfortable interiors. Passengers tend to use their travel time to rest or to work, so train interiors and their systems need to incorporate features that meet these requirements. Among the various systems that integrate train interiors, the flooring system is one of the systems with the greatest impact on passenger safety and comfort. It is also one of the systems that takes more time to install on the train, and which contributes seriously to the weight (mass) of all interior systems. Additionally, it presents a strong impact on manufacturing costs. The design of railway floor, in the development phase, is usually made relying on a design software that allows to draw and calculate several solutions in a short period of time. After obtaining the best solution, considering the goals previously defined, experimental data is always necessary and required. This experimental phase has such great significance, that its outcome can provoke the revision of the designed solution. This paper presents the methodology and some of the results of an experimental characterisation of composite panels for railway application. The mechanical tests were made for unaged specimens and for specimens that suffered some type of aging, i.e. heat, cold and humidity cycles or freezing/thawing cycles. These conditionings aim to simulate not only the time effect, but also the impact of severe environmental conditions. Both full solutions and separated components/materials were tested. For the full solution, (panel) these were: four-point bending tests, tensile shear strength, tensile strength perpendicular to the plane, determination of the spreading of water, and impact tests. For individual characterisation of the components, more specifically for the covering, the following tests were made: determination of the tensile stress-strain properties, determination of flexibility, determination of tear strength, peel test, tensile shear strength test, adhesion resistance test and dimensional stability. The main conclusions were that experimental characterisation brings a huge contribution to understand the behaviour of the materials both individually and assembled. This knowledge contributes to the increase the quality and improvements of premium solutions. This research work was framed within the POCI-01-0247-FEDER-003474 (coMMUTe) Project funded by Portugal 2020 through the COMPETE 2020.

Keywords: durability, experimental characterization, mechanical tests, railway flooring system

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