Journal of the Korea institute for structural maintenance and inspection
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v.25
no.4
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pp.56-64
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2021
Leakage due to deterioration and damage is one of the major causes of volume change by freezing and thawing, and it leads micro-cracking and surface scaling in concrete structures. The deterioration of damaged concrete accelerates with the chloride attack. Thus, in the detailed guidelines for facility performance evaluation (2020), the quality of cover concrete and the freeze-thaw (FT) repetition cycle were newly suggested for concrete durability assessment. The quality of cover concrete should be evaluated by the rebound hammer test and the FT repetition cycle should be also considered in the deterioration environmental assessment. This study suggested the application of fast dynamic based nonlinear ultrasound method to monitor initial micro-scale damage under freezing and thawing environment. Concrete specimens were fabricated with different water-cement ratios (40%, 60%) and air contents (1.5% and 3.0%). The compressive strength, rebound number, relative dynamic modulus, and nonlinear ultrasound were measured with different FT cycles. The scanning electron microscopy was also performed to investigate the micro-scale FT damage. As a result, both the rebound number and the relative dynamic modulus had difficulty to detect early damage but the proposed method showed a potential to detect initial micro-scale damage and predict the FT resistance performance of concrete.
This study was performed to investigate the changes of amount of S. typhimurium during cooking processes using pork and japchae (a Korean food which is made from meat, vegetables and noodles), and to support a practical application to develop a hazard analysis critical control point (HACCP) model. The pork was purchased in a retail shop, cut ($0.5\;cm\;{\times}\;10\;cm\;{\times}\;10\;cm$, 25 g), tested for Salmonella contamination (results: negative), inoculated with S. typhimurium ($10^{7}\;CFU/g$), then treated in various conditions related to cooking. Mter thawing for 24 hours in various conditions, the number of S. typhimurium was increased to $10^{10}\;CFU/g$ at a refrigerated temperature ($4~10^{\circ}C$), and to $10^{21}\;CFU/g$ at room temperature ($22~29^{\circ}C$). Mter thawing in a microwave oven for 40 seconds, the number of S. typhimurium increased to $10^{8}\;CFU/g$. During the thawing period, the number of S. typhimurium increased over time. At the refrigerated temperature, the number of the bacteria was $10^{10}\;CFU/g$ after 24 hours, $10^{13}\;CFU/g$ after 48 hours, and $10^{20}\;CFU/g$ after 72 hours. At room temperature the number of bacteria reached $10^{11}\;CFU/g$ in 2 hours, $10^{15}\;CFU/g$ in 4 hours, $10^{16}\;CFU/g$ in 8 hours, $10^{18}\;CFU/g$ in 12 hours, and $10^{21}\;CFU/g$ in 24 hours. Mter cooking in a frying pan (150{\pm}7^{\circ}C$) for 3 minutes, the bacterial count was $10^{16}\;CFU/g$. After cooking in hot water for 20 minutes, the bacterial count was $10^{7}\;CFU/g\;at\;60^{\circ}C,\;10^{6}\;CFU/g\;at\;63^{\circ}C,\;and\;10^{4}\;CFU/g\;at\;65^{\circ}C$. The fried pork was mixed with cooked vegetables, noodles, sesame oil, sesame seeds, and seasonings to make Korean japchae. This process took $10{\pm}2$ minutes. The bacterial count in the japchae increased to $10^{7}\;CFU/g$ from the count of $10^{6}\;CFU/g$ of the fried pork before it was mixed with the other ingredients. These results indicate that the amount of S. typhimurium is effected by various different cooking processes. This study can suggest that pork should be cooked in water at over $65^{\circ}C$ for 20 minutes in order to prevent food poisoning, if the pork is contaminated with S. typhimurium. The presence of S. typhimurium in the raw pork is identified in an HA for japchae, and the primary CCP for japchae is inadequate cooking (cooking method and time/temperature). We need to standardize time-temperature-size and amount of pork in cooking japchae, because pork is usually cooked in ordinary frying pans when we make this food.
Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps, freezing media and embryonic stages on the rates of viability and development of cryopreserved mouse embryos. Female ICR mice ($6{\sim}8$ weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only by cryoprotectant step (1 step${\sim}$4 step) but also in a variety of media (HTF, IVF medium, D-PBS) and cell stage. The results were as follows: There is no clear advantage in these freezing media of rapid method, but 4 cell and 8 cell of slow method (2, 3, 4 step) have advantage in D-PBS. The development of embryos according to cell stage become greater in 8 cell stage. In the treatment steps of cryopreservation, the development of embryo to blastocyst was similar among rapid method, but the development of 4 cell and 8 cell embryos to blastocyst according to slow method was better than rapid method.
Journal of the Korea institute for structural maintenance and inspection
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v.9
no.4
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pp.235-242
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2005
This study is on the evaluation of performance of polymer cement mortar which is used for pre-wetting spray method. Pre-wetting spray method is an epoch-making method to repair concrete structures damaged, which is added a small quantity water preciously to dry mortar to reduce dust and rebound and spray mortar mixed with fixed quantity water at nozzle before spray. The result showed that physical performance such like compressive, flexural and adhesive strength of polymer cement mortar, TS 100 used for pre-wetting spray method was superior to other repair mortar. Also durable performance such as resistance on permeability of chloride ion, carbonation, chemical and freezing-thawing was excellent.
The studies on the carried out to investigate the effective concentration of cryoprotectant agents and sucrose by one-step straw method of bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1$\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentrations of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as devellpmental rate on in vitro culture or FDA-test. The results are smmarized as followes : 1. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was attained 2.0M glycerol, 2.0M DMSO, 1M or 2.0M propanediol. 2. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was obtained single cryoprotectant(6.7~17.4%) than mixed cryoprotectants(6.7~16.7%). 3. In vitro developmental rate of bovine embryos after rapid frozen-thawing in the freezing medium added 0.25M and 0.50M sucrose were higher cleavage rate than those of sucrose concentration of 0.75M and 1.00M. 4. The freezing methods on in vitro developemental rates of bovine embryos was attained slow freezing method(9.70~15.6%) higher than rapid freezing method(9.4~13.3%).
This study was to test whether the viability of bovine hatched blastocysts (HBs) can be maintained after vitrification and thawing. The HBs were produced in vitro at Day 9 and Day 10 after IVF, and they were classified to small (S-HBs; ø$\leq$300 ${\mu}{\textrm}{m}$) and large(L-HBs; ø>300 ${\mu}{\textrm}{m}$) on the basis of embryo diameter using eyepiece micrometer. As freezing solution, we used EFS35 which consisted of 35% ethylene glycol (EG), 18% ficoll, 0.3 M sucrose and 10% FBS added in mDPBS. Vitrification was taken by two-step method, the HBs were equilibrated in 10% EG for 5 minutes and then shortly exposed in EFS35 and plunged into L$N_2$for 30~45 sec. After thawing, the survival rates were assessed by the re-expansion of the blastocoel during 2 h and 16 h of culture. The results obtained in these experiments were summarized as follows; 1) When the blastocysts(40.8%) recovered at Day 8 after IVF were further cultured for 24 h(Day 9 after IVF) and 48 h(Day 10 after IVF), the rates of HBs were 20.5% and 6.7%, respectively. Also, the total cell number of HBs on Day 9 was significantly higher than that of HBs on Day 10 (p<0.01). 2) When the effects of freezing solution to the survival of Day 9 L-HBs were examined, the rate of vitrified group (75.7%) was significantly lower than 100% of control and exposed group(p<0.05). 3) When the survival rates of vitrified HBs according to size and developmental age were examined, the data of L-HBs (75.5%) and S-HBs(63.6%) on Day 9 were slightly higher than those of L-HBs(64.3%) and S-HBs(60.7%) on Day 10. 4) Also, when the in vitro survival of Day 9 HBs was evaluated under different culture condition after thawing, the result in culture medium only (79.3%) was significantly higher than 43.2% in co-culture group (p<0.05). These results demonstrated that bovine HBs can be successfully cryopreserved by two-step vitrification method using EFS35.
Objective: The purpose of this study was to evaluate the efficacy and effect of various cryopreservation method on the survival and the cytoskeletal stability of metaphase II mouse oocyte. Methods: Mouse ovulated oocytes were collected and cryopreserved by a modified slow-freezing method with 1.5 M 1, 2-propanediol (PrOH)+0.1 M sucrose or by vitrification using cryo loop and EM grid with 40% ethylene glycol+0.6 M sucrose. Four hours after thawing, intact oocytes were fixed and stained with fluorescein isothiocyanate (FITC)-conjugated monoclonal anti-$\beta$-tubulin antibody to visualize spindle and propidium iodide (PI) to visualize chromosome. Spindle morphology was classified as follows: normal (barrel-shaped), slightly and absolute abnormal (multipolar or absent). Results: Survival rate of the frozen-thawed oocytes in vitrification group was significantly higher than that of slow-freezing group (62.7% vs. 24.4%, p<0.01). Vitrification with cryo loop showed significantly higher survival rate than that with EM grid (67.7% vs. 53.5%, p<0.05). On the other hand, proportion of normal spindle and chromosome configurations of the frozen-thawed oocytes between two vitrification group was not significantly different. Conclusion: For mouse ovulated oocytes, vitrification with cryo loop may be a preferable procedure compared to slow-freezing method. Further study should be needed to investigate developmental competency of frozen-thawed mouse oocytes.
Ha, Rim;Shin, Hyunjoo;Nam, Gibeom;Park, Sanghyun;Kang, Taegu;Song, Hyunoh;Lee, Hyuk
Journal of Korean Society on Water Environment
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v.32
no.6
/
pp.520-527
/
2016
Phycocyanin (PC) is one of the water-soluble accessory pigments of cyanobacteria species, and its concentration is used to estimate the presence and relative abundance of cyanobacteria. In laboratory experiments, PC content of field data were determined using Sarada's freeze-thaw method in algal bloom season. The effectiveness of three selected extraction methods (repeated freeze-thaw method, homogenization, power control) for PC were determined. The extraction efficiency of phycocyanin was the highest (of the methods compared) when a single freezing-thawing cycle was followed by pre-sonication. Applying this optimized method to surface water of Korean inland waters, the average concentration distribution was estimated at $2.9{\sim}51.9mg/m^3$. It has been shown that the optimized pre-sonication method is suitable to measure cyanobacteria PC content for the characterization of inland waters. The approach and results of this study indicates the potential of effective methods for remote monitoring and management of water quality in turbid inland waters using hyperspectral remote sensing.
Epididymal sperm cryopreservation provides a potential method for preserving genetic material from males of endangered species. This pilot study was conducted to develop a freezing method for tiger epididymal sperm. We evaluated post-thaw sperm condition using testes with intact epididymides obtained from a Siberian tiger (Panthera tigris altaica) after castration. The epididymis was chopped in Tyrode's albumin-lactate-pyruvate 1x and incubated at 5% CO2, 95% air for 10 min. The Percoll separation density gradient method was used for selective recovery of motile spermatozoa after sperm collection using a cell strainer. The spermatozoa were diluted with modified Norwegian extender supplemented with 20 mM trehalose (extender 1) and subsequent extender 2 (extender 1 with 10% glycerol) and frozen using LN2 vapor. After thawing at 37℃ for 25 s, Isolate® solution was used for more effective recovery of live sperm. Sperm motility (computerized assisted sperm analysis, CASA), viability (SYBR-14 and Propidium Iodide) and acrosome integrity (Pisum sativum agglutinin with FITC) were evaluated. The motility of tiger epididymal spermatozoa was 40.1 ± 2.0%, and progressively motile sperm comprised 32.7 ± 2.3%. Viability was 56.3 ± 1.6% and acrosome integrity was 62.3 ± 4.4%. Cryopreservation of tiger epididymal sperm using a modified Norwegian extender and density gradient method could be effective to obtain functional spermatozoa for future assisted reproductive practices in endangered species.
This study was carried out to investigate the effect of straw loading method and thawing protocol on the in vitro development of in vitro produced mouse blastocysts cryopreserved by vitrification. Three loading types of straw I, Il and III on loading and sealing method were made for vitrification. The ability of the solution on straw loading methods to remain vitreous during warming was tested by exposed in air for 1 to 10 s sec. and then plunged the vitrified straws into water bath at 25°C. Embryos to be vitrified were equilibrated to the 20% EG for 5min. and exposed in EFS 40 for 1min. The plug ends of Straw I and Straw II were sealed with straw powder and Straw III was treated straw powder, followed by heat sealing and then plunged into LN$_2$. The results obtained in these experiments were summarized as follows; 1) Straw I embryo column mostly changed from transparent to opaque upon thawing without exposure in air for 3-6 sec. Straw II embryo column was I improved partially but was not remained completely vitreous during warming. However, when Straw lll loading method was used, the embryo column was remained vitreous completely. 2) High survival rates and development rates of each groups (middle blastocysts and hatching blastocysts) of vitrified embryos were obtained by using Straw III loading method than Straw I method (P<0.05). And the range of s standard error was low in Straw lll method. 3) When the embryos vitrified-frozen were placed in air for 3, 5 and l0sec. and then warmed rapidly in water bath at 25$^{\circ}C$, the survival rates after 24h of culture were 72.7-87.1% and the development rates to hatching stage after 48h of culture were 34.0-48.4%. There were no significantly differences according to exposure time in air during warming. In conclusion, the present results showed that highly survival and low standard error of vitrified-frozen mouse bIastocysts were obtained by using straw lll loading, double sealing and appropriate 2 step warming method.
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