The cell culture of Angelica gigas Nakai producing decursin derivatives and immunostimulating polysaccharides was preserved in liquid nitrogen after pre-freezing in a deep freezer at -70$^{\circ}C$ for 480 min. The effects of the cryoprotectant and pretreatment before cooling were investigated to obtain the optimal procedure for cyropreservation. When compared to mannitol, sorbitol, or NaCl with a similar osmotic pressure, 0.7 M sucrose was found to be the best osmoticum for the cryopreservation of A. gigas cells. In the pre-culture medium, the cells in the exponential growth phase showed phase showed the best post-freezing survival after cryopreservation. A mixture of sucrose, glycerol, and DMSO was found to be an effective cryoprotectant and a higher concentration of the cryoprotectant provided better cell viability. When compared with the vitrification, the optimum cryopreservation method proposed in this study would seem to be more effective for the long-term storage of suspension cells. The highest relative cell viability established with the procedure was 89%.
PARK SEONG-JE;HONG YONG-JU;KIM HYO-BONG;KIM HYEON-JU;SHIN PHIL-KWON
Proceedings of the Korea Committee for Ocean Resources and Engineering Conference
/
2004.11a
/
pp.1-6
/
2004
This paper shows the study on the design and another applications of the spray-freeze dryer for the production of valuable material powders. Powder production and handling has been an integral part of material extracting processing and pharmaceutical processing because of the wide use of oral dosage forms. There are a few commonly used powder preparation methods including mechanical milling, precipitaion, spray drying, freeze drying, and so on. In general, methods available for preparing inhalation powders are limited due to certain inhalation powder's sensitive nature to the processing environments. This is particularly true for preparing dry powder aerosols where the aerodynamic particle size($<5{\mu}m$) and the size distribution are pivotal. Supercritical fluid antisolvent and spray freeze drying have recently emerged as promising techniques for producing powders for use in microcapsulation. However, the aerosol applications of these powders are yet to be explored. The purpose of this study is to test the feasibility of using spray freeze-dried valuable material powders for aerosolization.
To investigate effect of seeding on post-thaw motility and viability of canine spermatozoa, the semen from male dogs which had been proved to be fertile in the past were frozen and seeded during freezing process. Post-thaw motility and viability of canine sperm which were frozen and seeded were investigated according to different seeding temperatures of $-5^{\circ}C$, $-10^{\circ}C$, or $-l5^{\circ}C$ and also according to different concentration of glycerol of 2%, 5% and 10%. In addition, post-thaw motility of canine sperm frozen by direct freezing in a deep freezer or programmed freezing in a programmed cell freezer was investigated. Post-thaw motility of canine sperm was compared according to different seeding temperatures : The sperm seeded at $-5^{\circ}C$ showed considerably higher post-thaw motility than that of non-seeding, and that seeded at $-10^{\circ}C$, or $-l5^{\circ}C$, respectively, in 2% and 5% glycerol groups on both 2 and 7day after freezing(p<0.05). In 10% concentration of glycerol, the sperm seeded at each seeding temperature showed considerably higher post-thaw motility than that of non-seeding group on day 7 after freezing(p<0.01). Post-thaw viability of canine sperm was compared according to different seeding temperatures : The sperm seeded at $-5^{\circ}$ showed significantly higher post-thaw motility than that of non-seeding, and that seeded at $-10^{\circ}C$, or $-l5^{\circ}C$, in 5% and 10% glycerol groups on day 7 after freezing(p< 0.05). In comparison of post-thaw motility of canine sperm seeded according to different concentration of glycerol, 5% glycerol group and 10% glycerol group showed considerably higher post-thaw motility than 2% glycerol group without difference between those two groups in all seeding temperatures($-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}C$) on day 2 and 7 after freezing(p<0.01). In comparison of post-thaw viability of canine sperm seeded according to different concentration of glycerol, 5% glycerol group and 10% glycerol group showed the same considerably higher post-thaw viability than 2% glycerol group on each thawing day(p<0.01). The canine sperm frozen and seeded by programmed freezing method showed considerably higher post-thaw motility than that frozen by direct freezing method in all different seeding temperatures($-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}$). These results indicated that the higher post-thaw motility and viability was obtained in the spermatozoa seeded than that of non-seeding, that among different seeding temperatures of $-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}C$, the sperm seeded at $-5^{\circ}C$ showed higher post-thaw motility and viability than the other temperatures, also among different concentrations fof glycerol of 2%, 5% and 10%, the sperm frozen and seeded in 5% and 10% concentration of glycerol showed higher post-thaw motility and viability than that in 2% of glycerol, and that the sperm frozen and seeded by programmed freezing method showed higher motility than that by direct freezing method.
Background: Subcutaneous injection of 5% formalin into the hind paw of the rat produces a biphasic nociceptive response. The second phase depends on changes in the dorsal horn cell function that occur shortly after an initial C-fiber discharge, spinal sensitization, or windup phenomenon. This study was performed to investigate the role of glutamate during spinal sensitization. Methods: Sprague-Dawley rats weighing 200 to 250 g were used for this study. Under light anesthesia (0.5% isoflurane) the rats were segregated in a specially designed cage and $50{\mu}l$ 0.5% formalin was injected subcutaneously in the foot dorsum of right hindlimb. Forty minutes after the formalin injection, the rat was quickly decapitated and spinal cord was removed. The spinal segments at the level of L3 (largest area) was collected and stored in a deep freezer ($-70^{\circ}C$). The mRNA gene expression of N-methyl-D-aspartate receptor (NMDAR) and the metabotropic glutamate receptor subtype 5 (mGluR5) were determined by the polymerase chain reaction. Results: The number of flinches was $19.8{\pm}2.3/min$. at one minute after formalin injection and decreased to zero after then. The second peak appeared at 35 and 40 minutes after formalin injection. The values were $17.8{\pm}2.2$ and $17.2{\pm}3.0/min$. The mRNA gene expressions of NMDAR and mGluR5 were increased by $459.0{\pm}46.8%$ (P < 0.01) and $111.1{\pm}4.8%$ (P > 0.05) respectively at 40 minutes after formalin injection. The increased rate of NMDAR was significantly higher than that of mGluR5 (P < 0.01). Conclusions: From these results it suggested that NMDAR partly contributed to the mechanism of central sensitization after the formalin test but mGluR5 did not.
Kim, Yong-jun;Park, Young-jae;Kim, Byeong-jin;Yu, Il-jeoung
Korean Journal of Veterinary Research
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v.34
no.4
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pp.851-855
/
1994
The semen from four male dogs which had been proven to be fertile in the past were frozen in a deep freezer at $-60^{\circ}C$ by simple freezing method using methanol and preserved at the same teperature for from 7 to 10 days. The semen were inseminated to 7 female dogs in estrus to find out the usability of this freezing method in artificial insemination for dogs. In addition, post-thaw motility and viability of sperm from two male dogs which had been fertile were also evaluated to investigate individual difference. Successful pregnancy was obtained by artificial insemination with canine semen frozen at $-60^{\circ}C$ by simple freezing method using methanol, namely, 3 bitches among 7 bitches which had been inseminated delivered puppies(42.8%). The average litter size of the whelping dogs were 4.3 puppies. The average post-thaw motility of canine sperm in the cases of conception was showen higher than those of non-conception(65.0% vs. 42.5%), along with the, same result in the average post-thaw viability between the two groups(53.3% vs, 27.5%). Individual difference of post-thaw motility and viability was obtained between two fertile dogs(p<0.05).
In the specimen of free PSA in the low concentration, the result in % bias from our institution and comparable evaluation institution was -33.7% which is exceeded % bias ${\pm}20%$ ; however, it was the domestically allowable limit recommended by the laboratory accreditation commission for specimen at the low concentration. In this paper, the cause was accredited by instability of free PSA substance within the specimen, and the specimen stability test was performed according to CLSI documents GP29-A2. After the low and high concentration specimen were made, and rapidly cooled down in a deep freezer with $-30^{\circ}C$, serum of two concentrations was measured for 10 consecutive days with 3 times a day by Architect i2000 and observed a change in the mean value. As the results of two groups, there were changes in the established target value, and a change level was evaluated by calculating it with % bias. The low concentration specimen had no significant reduction until the 4 day lapse in cold storage. However, % bias were reduced by -17.5% from the 5 day lapse, by 21.5% after the 7 day lapse, and by -26.9% after the 9 day lapse. The frozen specimen had only intra-day variation for 10 days. In the high concentration specimen, bias began to show as -12.2% from the 3 day lapse in cold storage. There was reduction by -28.9% from the 5 day lapse, by -39% after the 7 day lapse, and by -42.9% after the 9 day lapse. In the frozen specimen, there was only intra-day variation like the low concentration specimen in cold storage.
BACKGROUND: Spoilage fungi can reduce the shelf life of fresh fruits and cause economic losses by lowering quality. Especially, mulberry fruits have high sensitivity to fungal attack due to their high water content (> 70%) and soft texture. In addition, the surface of these fruits is prone to damage during harvesting and postharvest handling. However, any study on postharvest spoilage fungi in mulberry fruit has not been reported in Korea. This study aimed to examine the spoilage fungi occurring in mulberry fruits during storage after harvest. METHODS AND RESULTS: In this study, we isolated postharvest spoilage fungi from mulberry fruits stored in refrigerator (fresh fruits) and deep-freezer (frozen fruits) and identified them. In the phylogenetic analysis based on comparisons of the ITS rDNA sequences, the 18 spoilage fungi isolated from mulberry fruits and the 25 reference sequences were largely divided into seven groups that were subsequently verified by high bootstrap analysis of 73 to 100. Alternaria spp. including A. alternate and A. tenuissima, were the most frequently isolated fungi among the spoilage isolates: its occurrence was the highest among the 18 isolates (38.9%). CONCLUSION: The findings of this study will be helpful for increasing the shelf life of mulberry fruits through the application of appropriate control measures against infection by spoilage fungi during storage.
Kim, Da Hae;Kim, Sungkyoon;Lee, Seungho;Choi, Yoon-Hyeong;Kim, Jin Hee
Journal of Environmental Health Sciences
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v.47
no.3
/
pp.205-226
/
2021
Objectives: Biomonitoring programs have been widely implemented in the field of environmental health, both in Korea and worldwide. Recently, it has been suggested that the storage, management, and utilization of biosamples collected from biomonitoring programs should be organized based on a biobank system. Therefore, we attempted to review the current status of representative biomonitoring programs and biobank systems that have been implemented in Korea and in other countries. Methods: We searched for bio-samples collected in domestic and foreign biomonitoring programs and their applications. For this, we referred to research papers, homepages hosted by biomonitoring programs, and project reports. We also checked information for biobanks related with biomonitoring programs, including the operating systems, facilities, technologies, and regulations of biobanks. Results: We summarized six domestic and 32 foreign biomonitoring programs. These biomonitoring programs collected bio-samples to determine the relationship between environmental chemicals and diseases. Domestically, bio-samples from KoNEHS, KorSEP, MOCEH, KoCHENS, and KorEHS-C were stored at -80℃ in a deep freezer at the National Institute of Environmental Research, while KNHANES samples were stored at Korea Biobank, which has a stabilized biobanking system with a well-established database. Nine foreign biomonitoring programs (JECS, China-NHBP, CKB, CHMS, NHANES, GerES, Germaan ESB, MoBa, and UK Biobank) were ongoing for large populations. Among them, CKB, GermanESB, and UK biobank have been maintained for at least 10 years with their own biomonitoring programs as well as advanced systems for the safe storage of bio-samples. Conclusion: Currently on-going biobanks have devoted considerable efforts to managing bio-samples for public purposes. The preceding domestic and foreign biomonitoring programs and biobanks will be great references for constructing biobank facilities and systems for environmental public health in Korea in the future.
In this study, we have examined the effects of the storage time and temperature on DNA quality and have also studied the effects of the hydration buffer in which DNA is dissolved. This study was performed using 160 human blood samples collected with informed consent from 2007 to 2008 in the hospital where this cohort study was performed. The DNA extracted was dissolved using distilled water (DW) or Tris-EDTA (TE) buffer, and stored in the deep freezer or refrigerator for up to 10 weeks at $-70^{\circ}C$, $-20^{\circ}C$, $4^{\circ}C$, and $25^{\circ}C$, respectively. DNA integrity was determined by the degree of smearing of DNA on the gel. After four weeks, all of the 20 DNA samples dissolved in DW and stored at $25^{\circ}C$ were entirely degraded. After 10 weeks, 6 of the 20 DNA samples dissolved in TE buffer and stored at $25^{\circ}C$ were fairly degraded, and 4 of the 20 DNA samples dissolved in DW and stored at $4^{\circ}C$ were fairly degraded. The 20 DNA samples dissolved in TE buffer and stored at $4^{\circ}C$ were stable for 10 weeks. DNA samples stored at $-20^{\circ}C$ and $-70^{\circ}C$ did not appear to degrade in either DW or TE buffer, even at the 10-week point. We suggest that TE buffer should use for DNA elution, in order to protect against degradation and to preserve DNA for a long period of time, and the samples should be stored at $-20^{\circ}C$ or $-70^{\circ}C$.
This study was conducted to examine the effects of recombinant Bovine Somatotropin (rBST), vitamin E(Vit. E) and selenium (Se) administration on the blood chemical values and hormone concentrations of serum in Hanwoo sires. Hanwoo sires were randomly assigned to five groups; 1) control, 2) rBST,0.09mg/kg body weight (BW), 3) Vit E, 1,500IU/kg BW, 4) Se 0.1mg/kg BW, 5) Vit E, 1,500IU plus Se 0.1mg/kg BW. rBST, Vit. E and Se for each experimental group were given 6 times at 15 days interval by intermuscular injection. Blood samples were collected ten times for experimental periods and separated the serum by centrifugation and stored into deep freezer. The concentration of albumin was the highest in Se group than those of any other groups (P<0.05) and Vit. E plus Se group was significantly higher than in the control and rBST groups (P<0.05). The concentrations of blood urea nitrogen (BUN) and creatinine were significantly higher in rBST group than any other groups (P<0.05). The concentration of total protein in rBST, Se and Vit. E plus Se groups were significantly higher than in control group (P<0.05). The concentrations of calcium, cholesterol,, glucose, inorganic phosphorus and triglycerides in serum were not difference in all experimental groups (P>0.05). The concentration of estradiol was slightly higher in Se and Vit. E plus Se groups than those of any other groups, but not significantly difference among the experimental groups (P<0.05). The concentration of testosterone was not affected by the administration of rBST, Vit. E and Se.
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