• Journal of Pharmaceutical Investigation
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• v.25 no.3
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• pp.249-264
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• 1995

Although liposome has many advantages as a pharmaceutical dosage form, its application in the industrial field has been limited because of some problems such as preparation method, reproducibility, scale-up, stability and sterilization etc. Liposomes prepared by microemulsification method had defined size, narrow size distribution, reproducibility and high entrapment efficiency. For enhancing the stability, the dry form of liposome was recommended. These types of liposome are proliposome and freeze-dried liposome. The liposome must have some properties for preparing of freeze-dried liposome; small size $(50{\sim}200\;nm)$, narrow size distribution and cryoprotectant. In this experiment, the liposomes containing 5-Fluorouracil(5-FU) and its prodrug(pentyl-5-FU-1-acetate; PFA, hexyl-5-FU-1-acetate; HFA) were made with soybean phosphatidylcholine, cholesterol, stearylamine(SA) and dicetyl phosphate(DCP) employing hydration method or microemulsification method using $Microfluidizer^{TM}$. Both or liposome types were MLV and MEL. After preparation, freeze drying and rehydration were performed. In the process of freezing, trehalose(Tr) was added as a cryoprotectant. Their evaluation methods were as follows; entrapment efficiency, mean particle size and size distribution, dissolution test, retain of entrapment efficiency and turbidity after freeze-drying. The results are summarized as belows. The entrapment efficiency of 5-FU was dependent on total lipid concentration and cholesterol content but that of PFA and HFA was decreased when cholesterol was added. When DCP and SA were added, entrapment efficiency was decreased. As the partition coefficient of drug was increased, entrapment efficiency was increased. Under the same condition, entrapment efficiency of MEL is similar to that of MLV. The mean particle size and size distribution of MEL were smaller than those of MLV. Dissolution rates of drug from both liposome types were comparatively similar. Dissolution rates of drugs with serum and liver homogenate were faster than without these material. After preparation of liposome, free drug was removed efficiency by Dowex 50W-X4. When liposome was freeze-dried and then rehydrated in the presence of Tr, characteristics of liposome were maintained well in MEL than MLV. Tr Was used successfully as a cryoprotectant in the process of freeze drying and the optimal ratio of Tr:Lipid was 4:1(g/g).

• ALGAE
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• v.21 no.1
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• pp.125-131
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• 2006

Long-term preservation of bloom-forming cyanobacteria was evaluated using cryopreservation and freeze-drying of nine strains belonging to four genera and seven species. All test strains, except Aphanizomenon flos-aquae NIER- 10028, showed partial or complete survival following cryopreservation and freeze-drying. Frozen and freeze-dried strains were preserved for more than two years and were revived monthly. Most strains showed higher post-thaw viability after cryopreservation, especially without cryoprotectant compared to freeze-drying. Microcystis aeruginosa NIER-10010, M. viridis NIER-10020, M. ichthyoblabe NIER-10023, M. novacekii NIER-10029 and Oscillatoria sancta NIER-10027 were revived after 2.5 years of cryopreservation. These results suggest that cryopreservation may be an easy and timesaving long-term preservation method for bloom-forming cyanobacteria.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.3
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• pp.202-206
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• 2000

The genetically-engineered Escherichia coli strain, DPD2540, which contains a fabA:::luxCDAbefusion gene, gives a bioluminescent output when membrane fatty acid synthesis is needed. For more pactical application of this strain in the filed as biosensor, freezedrying was adopted. A 12% surcrose solution with Luria-Bertani (LB) broth, as determined by the viability after freeze-drying, was found to be most most effective composition for lyophilization solution among various compositions testitons tested. Rapid freezing with liquid nitrogen also gave the best viability after freeze-drying as compared to samples frozen at-7$0^{\circ}C$ and -2$0^{\circ}C$. The biosensing activities of the cells showed a greater sensitivity when the cells from the expontial phase were freeze-dried. Finally, the optimum temperature for use of the freeze-dried cells in the biodencor field was determined.

• Korean Journal of Food Science and Technology
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• v.54 no.2
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• pp.224-227
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• 2022

In this study, the protective effects of turanose on Saccharomyces cerevisiae (Baker's yeast) were examined during the freeze-drying process to evaluate the feasibility of utilizing turanose as a novel cryoprotectant. The survival rate of the Baker's yeast cells improved substantially with a cryoprotective medium containing turanose in a substitution-dependent manner. In accordance with these survival rates, the yeast cell surfaces became smoother as the turanose content increased. Turanose with skim milk maintained the viability of the Baker's yeast, which improved substantially upon storage at -20℃. Thus, it is thought that turanose will exhibit excellent preservation effects during the distribution of Baker's yeast. Finally, these results suggest that turanose has the potential to be used as a novel cryoprotectant against various microorganisms.

• The Korean Journal of Food And Nutrition
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• v.14 no.5
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• pp.441-445
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• 2001

For the developement of probiotics, three strains haying psychorophilic and salt tolerant characteristics were isolated from Kimchi. Among the isolated strains, MGl9, MG89 and MG208 were identified as Lactobacillus brevis, Enterococcus faecium and Lactobacillus plantarum, respectively. The relationship between cryoprotectant and the viability of freeze dried strains has been studied. The most effective cryoprotectant for MGl9 was found to be 10% skim milk contained 1% soluble starch haying the survival rate of 71.4%. In MG89, 10% skim milk contained 1% sucrose and 10% skim milk contained l% fructose in MG208 were showed effective cryoprotectant having the survival rate 68.9% and 64.7%. respectively These results suggest that these isolated strains can play an important role as probiotics in aquaculture.

• Food Science and Preservation
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• v.22 no.2
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• pp.167-173
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• 2015

This study was conducted to evaluate effect of adding cryoprotectant agents on the growth of lactic acid bacteria during storage of powdered Kimchi. Powdered Kimchi was prepared by adding 1.5% cryoprotectant (glucose, maltose, lactose, and sucrose) and freeze-dried. For the preparation of micro-sized particle of Kimchi powder, the freeze-dried Kimchi was powered at 14,000 rpm for 2 min. The survival ratio of lactic acid bacteria in the powdered Kimchi was monitored during storage period of 4 months at -20, 0, 4, and $25^{\circ}C$ after the capsulation of the powedered Kimchi. The number of lactic acid bacteria in the powdered Kimchi capsule was the greatest stored at $-20^{\circ}C$, and the addition of glucose in cryoprotectant showed higher survival rate of lactic acid bacteria than that of control. More than $10^7CFU/g$ of lactic acid bacteria were survived in the powdered Kimchi stored at 0 and $4^{\circ}C$. However, the lactic acid bacteria were not detected in the powdered Kimchi stored at $25^{\circ}C$. As a results, the addition of cryoprotectant agents in the manufacturing process improved the survival rate of lactic acid bacteria in powered Kimchi products with accompanying with a cold-chain system for the distributon of powdered Kimchi products.

• Fisheries and Aquatic Sciences
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• v.8 no.1
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• pp.10-16
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• 2005

Conditions for sufficient and rapid distribution of a cryoprotectant (sorbitol solution) into intact fish muscle (Oncorhynchus mykiss) were studied as changing in the residual Ca2+ ATPase activity during freeze/thaw cycling. Chunks of the fish muscle were immersed in 4 concentrations of sorbitol solutions ($20\%$, $30\%$, $45\%$, and $60\%$) by a shaker mechanism at 5$^${circ}C. Whole immersion samples (W) showed a higher value of the residual Ca2+ ATPase activity than those in the untreated controls (C), except in the treated controls (TC), while less effect of immersion concentration could be found. Comparing the extent of penetration of sorbitol into the surface layer to inner layer of immersed fish chunks, outer portion samples achieved excellent cryoprotection with $100\%$ of the residual ATPase activity values or more. For the inner portion samples, $30\%$ and $45\%$ sorbitol solution treatments indicated a higher ATPase activity than $60\%$ treatment. At high concentrations, mass transfer rates during osmotic dehydration might berapid and it causes faster surface drying by dewatering at surface solute layer. Periodically immersed and relaxed samples, W (5-3-1), led to good cryoprotection effect: W (5-3-1) indicated high residual Ca2+ ATPase activity values and the residual ATPase activity values excess $100\%$ in immersion of $30\%$ and $45\%$ sorbitol solutions.

• Journal of Embryo Transfer
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• v.29 no.1
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• pp.51-57
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• 2014

The objectives of this study were to investigate the effects of trehalose as a cryoprotectant for porcine freeze-dried spermatozoa, to find the optimal freeze-drying time and storage periods of freeze-dried spermatozoa, and to find out pronuclear formation rates, cleaved rates, and embryo development through intracytoplasmic injection of freeze-dried spermatozoa on porcine oocytes. The survival rates of spermatozoa after freeze-drying with trehalose treatment were significantly higher than those of them without trehalose treatment (p<0.05). The highest survival rates were found at 75 mM trehalose treatment. The longer storage periods after freeze-drying seemed to have a lower survival rates. Development in culture of pig by ICSI with trehalose treatment were significantly higher than those of them without trehalose treatment (p<0.05). Shorter freeze-drying time of spermatozoa was resulted in the highest cleaved rates and embryo development.

• The Korean Journal of Food And Nutrition
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• v.17 no.3
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• pp.286-293
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• 2004

Extract of Eleutherococcus senticosus, Lycium chinensis, Angelica acutiloba and Schizandra chinensis were investigated to determine whether its addition may inflict on growth of lactic acid bacteria, freeze dry and cell viability during the storage. In cultivation with herbal extract, all strains tested did not demonstrate a significant reduction in their cell population, particularly Lactobacillus. acidophilus, L. bulgaricus, L. paracasei. But Schizandra chinensis extract inhibited growth of several strains. Immediately after freeze-drying using the skim milk 10%(w/v), sucrose 5%(w/v) with herbal extract 2.5%(v/v), the percentage viability was about 81.7%, 63.8%, 73.2%, 78.1 % in L. acidophilus, L. bulgaricus, L. paracasei, L. casei respectively. The protective effect of herbal extract to cell damage from freeze-drying was weak in comparison with control. During accelerated storage of freeze-dried lactic acid bacteria, those survival rate decrease rapidly, reaching 8 -18% in one month. But addition of Eleutherococcus senticosus extract in freeze dry of L. acidophilus showed a positive activity in storage.

• KSBB Journal
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• v.30 no.3
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• pp.109-113
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• 2015

Current drying and encapsulation methods for probiotics manufacturing are complicate and cost-burdened processes. The aim of this study was to develop a simple ingredient mixture to make probiotic granules via one-step process, providing not only a cryoprotective effect during freezing and drying but also high survival ratio in gastrointestinal tract. As cryoprotectans, commercially available ingredients including skim milk, monosaccharide (trehalose or glycerin), maltodextrins (with low or high degree of equivalents) were used. Their cryoprotective effect during lyophilization and survival ratios in artificial gastric juice and bile salt were measured against 3 strains of lactic acid bacteria (LAB) (Lactobacillus plantarum, Lb. brevis, and Lactococcus lactis). As results, 3 mixtures with different compositions showed a cryprotective effect on LAB tested and the best compostion was dependant upon LAB; skim milk 10%, trehalose 15%, glycerin 0.5%, and NaCl 1% was for Lb. plantarum and Lc. lactis, and maltodextrin 10% instead of skim milk was for Lb. brevis. In addition, those mixtures showed similar survival effect on LAB tested. These results demonstrate that skim milk or maltodextrins with trehalose, glycerin, and NACl can be effectively used for onestep lyophilization of LAB as an alternative method of encapsulation.