• Food Science of Animal Resources
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• v.24 no.2
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• pp.151-155
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• 2004

This study was performed to develop the method of differentiation fresh and frozen meat by using the measurement of mitochondrial malate dehydrogenase. The principle of this experiment is based on the fact the enzyme proteins associated with mitochondria membrane could be released by freezing. The methods were studied by measurements of protein concentration of meat press juice, WHC (water-holding capacity), drip loss and mitochondrial malate dehydrogenase enzyme activity. Samples were stored at 4$^{\circ}C$ and -18$^{\circ}C$ during storage period, respectively. Protein concentration of meat press juice was ranged from 8.5 mg/mL to 12.7 mg/mL and increased by freezing below at -18$^{\circ}C$(p<0.05). The WHC was not significantly different between fresh meat and frozen chicken meat (p＞0.05). The amount of drip loss of fresh and frozen chicken meat at 4$^{\circ}C$ and -18$^{\circ}C$ was not significantly different (p＞0.05). Mitochondrial malate dehydrogenase activity of frozen meat (-18$^{\circ}C$) was significantly higher (p＜0.05) than that of fresh meat. Also, enzyme activity of frozen meat was maintained at the same level after 3 minutes reaction. But fresh meat had not this reaction. From these results, it suggests that mitochondrial malate dehydrogenase can be used as a promising enzyme to differentiate between fresh and frozen meat.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.14 no.8
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• pp.4033-4038
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• 2013

Sedimentary rocks from construction waste are discarded through open storage and landfilling, which causes an increase in construction cost and inefficient of execution of works. Some sandstone are selected and utilized as aggregates, but shale is buried as industrial waste. Therefore, in this research, we evaluated weathering properties of shale aggregate that is widely distributed throughout Daegu-Kyeongbuk region and freeze-thaw characteristics of concrete according to the replacement ratio of shale aggregate, in an effort to stabilize aggregate supply-demand in Daegu-Kyeongbuk region and develop alternative aggregates. We used red shale and black shale in the experiment, which were exported from a construction site in Deagu. We verified the usage of shale as a concrete aggregate by comparing andesite, which is broadly used as a thick aggregate for concrete, to hornfels, which is a metamorphic sedimentary rock. As a result of the experiment, we observed no degradation phenomenon for andesite and hornfels. However, a part of country rock containing black shale was found to be exfoliated. Red shale started having cracks in the direction of stratification after 1.5 months of direct exposure, and it broke into smaller pieces after approximately 4 months. After 300 cycles of freeze-thaw process on the concrete manufactured according to the replacement ratio of shale aggregate, the modulus of elasticity was 97% for plain and 95% for hornfels. In the case of RS_100, it was 57% after 210 cycles, and for BS_100, it was 54% after 240 cycles. Therefore, we established that, as the number of repetition increases, the freeze-thaw resistance decreases dramatically.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.421-426
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• 1986

This study was attempted to find out effective storage methods of Lactobacillus casei YIT 9018, industrial strain for fermented mil k production, without severe bacterial death and activity deteriorations. The cryoprotective effect of the ammo acid mixture consisting of glycine and DL-g1utamic acid on the test strain were examined and also compared with those other protectants already reported. The apparent protective effect by the amino acid mixture was observed to controls. Both glycine and DL-glutamic acid prevented the freezing death of test strain and his effect of 1. casei YIT 9018 had reached stationary stage in MRS-broth 18h after inoculation. Cells harvested from stationary stage were most resistant to freezing damage. The viability of the test strain was affected by rehydration media and the recovery of viable cells was increased about threefold when amino acid mixture was used for rehydration. The presence of non-fat milk solid (NFMS), sucrose and lactose in amino acid mixture increased viability of the test strain up to 85％. In this case, optimal concentrations of NFMS, sucrose and lactose were 10％, 7.5-10％, 7.5-10％, respectively.

• Archives of Plastic Surgery
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• v.36 no.2
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• pp.127-134
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• 2009

Purpose: Adipose tissue injection as a free graft for the correction of soft - tissue deficiency or depression deformity is a widespread procedure in plastic surgery. This study is to analyze the changes and viability of cryopreserved adipose tissue and to find out efficient long - term storage period. Methods: After centrifugation of aspirated abdominal tissues, $10m{\ell}$ of packed Adipose tissue were freezed at $-20^{\circ}C$. For 2, 4, 6, 8 months, each frozen samples were taken and injected into scalp of SCID mice. After 15 weeks, injected Adipose tissue were sampled and analyzed at 2 months interval. We compared and analyzed each group about the weight of the injected fat, histologic impressions, activity of mitochondria, size of a fat cell and rate of survival. Results: Significant weight changes were observed in cryopreservation for 2 months(p<0.05). Histologic changes were observed, independent of the freezing period with H - E stain. Among cryopreservations for 2, 4, 6 months, no significant change were observed. The reduction of mitochondrial enzymatic activity was observed independent of time interval but activity of mitochondrial dehydrogenase was reduced less than 50% in MTT assay. Conclusion: Freezing in $-20^{\circ}C$ for 6 months has no adverse effect to Adipose tissue, but fragile adipocytes, damaged cell membrane during harvesting procedure, were disrupted within 1 - 2 month and the maximum volume reduction were followed less than 2 months. These results demonstrate that tissue preparation cells without membrane damage have the greatest viability level and cryopreservation less than 2 months has great volume effect and cryopreservation for 6 months has stable volume effect.

• Korean Journal of Food Science and Technology
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• v.42 no.5
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• pp.565-570
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• 2010

A coenzyme Q10 nanoemulsion was prepared using high pressure homogenization with different valve type conditions (A, B, and C) and cycle numbers (1, 2, and 3). The particle size, transmittance, zeta potential, and coenzyme Q10 content of the prepared coenzyme Q10 nanoemulsion were measured. The stability of the prepared coenzyme Q10 nanoemulsion was evaluated on heating ($95^{\circ}C$), freezing ($-20^{\circ}C$), and different pH (2-10) conditions. Also, the prepared coenzyme Q10 nanoemulsion was stored at different temperatures of 4, 25, and $40^{\circ}C$ for 12 weeks to evaluate its storage stability. In this study, the optimal conditions of high pressure homogenization for the preparation of a coenzyme Q10 nanoemulsion were identified to be 150 MPa, C valve, and a cycle number of 3. The results showed that the prepared coenzyme Q10 nanoemulsion had an average particle size of 40 nm, generated no deposits or floating matter when stored at either 4 or $25^{\circ}C$ for 12 weeks, and displayed excellent dispersibility and transparency when processed at different pHs (4-10) or heating ($95^{\circ}C$) and, freezing ($-20^{\circ}C$) conditions. Our results indicated that a coenzyme Q10 nanoemulsion prepared by high pressure homogenization can be used for preparing beverages in the food industry.

• Korean journal of food and cookery science
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• v.14 no.4
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• pp.394-399
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• 1998

The juice of garlic (Euichun variety) was extracted and concentrated by heating at 90$^{\circ}C$, by using a rotary vacuum evaporator at 45$^{\circ}C$, or by freezing at -50$^{\circ}C$ until the volume was reduced to 70% of the original's. The concentrated garlic juice was packed into 15 ml test tubes wrapped with aluminum foil and kept at 4$^{\circ}C$ or 25$^{\circ}C$ for 60 days. Changes of browning, microbiological and sensory characteristics of the concentrated garlic juices were monitored every 10 days. The specific gravity and viscosity of the prepared juices decreased in the juices concentrated at 90$^{\circ}C$, 45$^{\circ}C$ and -50$^{\circ}C$ in order. Browning of the concentrated garlic juices was slower during the storage at 4$^{\circ}C$ than at 25$^{\circ}C$. Browning occurred rapidly in the juice concentrated at 45$^{\circ}C$ during the storage, especially at 25$^{\circ}C$. The numbers of mesophilic and psychrotrophic bacteria in the juices did not increase significantly during the storage, which means the garlic juices had good shelf-life. The CFUs/ml of garlic juice concentrated at 90$^{\circ}C$ were lower about 1 to 2 log cycles than those in other concentrated juices. The juice concentrated at 90$^{\circ}C$ showed the weakest garlic odor and the strongest cooked odor among the juices. The juice concentrated at -50$^{\circ}C$ had the freshest odor, especially stored at 4$^{\circ}C$, but the juice concentrated at 90$^{\circ}C$ had lowest score in fresh odor. Brown color was dark in the juice concentrated at 45$^{\circ}C$ and green color of all the juices did not change significantly during the storage.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.2
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• pp.1200-1206
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• 2015

The major target for drug discovery, G-protein coupled receptor (GPCR) is involved in many physiological activities and related to various diseases and disorders. Among experimental techniques relating to the GPCR drug discovery process, various cell-based screening methods are influenced by cell conditions used in the overall process. Recently, the utilization of frozen cells is suggested in terms of reducing data variation and cost-effectiveness. The aim of this study is to evaluate various conditions in cell freezing such as temperature conditions and storage terms. The stable cell lines for calcium sensing receptor and urotensin receptor were established followed by storing cultured cells at $-80^{\circ}C$ up to 4 weeks. To compare with cell stored at liquid nitrogen, agonist and antagonist responses were recorded based on the luminescence detection by the calcium induced photoprotein activation. Cell signals were reduced as the storage period was increased without the changes in $EC_{50}$ and $IC_{50}$ values $EC_{50}:3.46{\pm}1.36mM$, $IC_{50}:0.49{\pm}0.15{\mu}M$). In case of cells stored in liquid nitrogen, cell responses were decreased comparing to those in live cells, however changes by storage periods and significant variations of $EC_{50}/IC_{50}$ values were not detected. The decrease of cell signals in various frozen cells may be due to the increase of cell damages. From these results, the best way for a long-term cryopreservation is the use of liquid nitrogen condition, and for the purpose of short-term storage within a month, $-80^{\circ}C$ storage condition can be possible to adopt. As a conclusion, the active implementation of frozen cells may contribute to decrease variations of experimental data during the initial cell-based screening process.

• Journal of Pharmaceutical Investigation
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• v.37 no.2
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• pp.79-84
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• 2007

This study was aimed to establish analytical method of Bi to develop a guideline of the bioequivalence test of tripotassium dicitrato bismuthate (TDB). For this purpose, a simple, specific and sensitive inductively coupled plasma-mass spectrometry (ICP/MS) method were developed and validated in human plasma. Various concentrations of bismuth standard solution (0-25ng/mL) were prepared with distilled water and human blank plasma. To 10mL of the volumetric flasks, 2mL of blank plasma was added with 8ml of distilled water. Bi standard solution was added to prepare the calibration samples and injected into ICP-MS. The plasma samples obtained from volunteers given 3 tablets of bismuth (total 900mg as TDB) were analyzed as described above. As a result, the coefficients of variation were <20% in quantitation limit (0.2 ng/mL) and <15% at the rest of concentrations. The stability test by repeated freezing-thawing cycles showed that the samples were stable only for 24hr. The stability tested for samples with a short-term period of storage at room temperature and pre-treatment prior to the analysis showed very stable over 24hr. In 8 healthy Korean subjects received Denol tablets at the dose of 900mg bismuth, AUC, $C_{max},\;T_{max}$ and half-life $(t_{1/2})$ were determined to be $198.33{\pm}173.78 ng{\cdot}hr/mL,\;64.48{\pm}27.06 ng/mL,\;0.52{\pm}0.21 hr,\;and\;5.15{\pm}2.67 hr$, respectively, from the plasma bismuth concentration-time curves. In conclusion, the method was suitable for the determination of bismuth in human plasma samples and could be applied to bioequivalence test of bismuth tablet.

• Microbiology and Biotechnology Letters
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• v.14 no.6
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• pp.433-439
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• 1986

Abundant amount of Chiamydia trachomatis could be amplified in mammalian McCoy cells and purified using descontinuous Uroarafin gradient centrifugation. As a chemical means io increase the Chlamydia trachomatis inclusions in McCoy cells IUdR treatment was found to be more effective than the cycloheximide treatment and was recommendanble for the proliferation of Chlamydia trachomatis. Centrifugation promoted Chlamydia trachomatis adhesion to McCoy cell surface, and maximal percentage of infected cells was obtained at about 3000g. The purified Chlamydia trachomatis could be kept in SPG solution for 48 hours at +4$^{\circ}C$ but for longer storage freezing to -7$0^{\circ}C$ was necessary.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.1
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• pp.81-87
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• 1991

There studies were carried for evaluation of the efficiency of freezing of hamster oocytes for use in a human sperm penetration assay. The hamster oocytes fully equilibrated in various cryoprotectant agents and inseminated with human sperm. After insemination with hamster oocytes, there was no difference in penetrated rates. Cumulus free oocytes equilibrated in 1.5M various cryoprotective agents and slowely cooled to temperature $-30^{\circ}C$ before rapid cooling and storage in nitrozen tank. After rapid thawing, survival rates of frozen oocytes according to cryo-protective agents were examined and the human sperm penetration assay with zona free hamster oocytes was conducted. 1. Survival rates of oocytes after cryoprotectants exposure have no significant difference (range 88-91%) and peneration rate was 51.1%. 2. Recovery and survival rate of frozen-thawed oocytes were 85.1 and 66.8%. There was no significant difference on cryoprotective agents. 3. Penetration rates of the frozen-thawed and intact oocytes were 69.0 and 77.0%, respectively. 4. Hamster oocytes cryopreservation provides a convenient way of supplying and trans-porting hamster oocytes for the assessment of the fertilizing potential of human spermatozoa.