• Applied Microscopy
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• v.30 no.4
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• pp.347-355
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• 2000

Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to some metalloproteins and nucleic acids, loosely bound to some metallothioneins or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors and transcription factors, thus rolls need accurate homeostasis of zinc ions. Zinc is an essential catalytic or structural element of many proteins, and a signaling messenger that is released by neural activity at many central excitatory synapses. Growing evidences suggest that zinc may also be a key mediator and modulator of the neuronal death associated with transient global ischemia and sustained seizures, as well as perhaps other neurological disease stoles. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartment ('vesicular zinc') which can be evidenced using histochemical techniques. Substances giving a bright colour or emitting fluorescence when in contact with divalent metal ions are currently used to detect them inside cells; their use leads to the so called 'direct' methods. The fixation and precipitation of metal ions as insoluble salt precipitates, their maintenance along the histological process and, finally, their demonstration after autometallographic development are essential steps for other methods, the so called 'indirect methods'. This study is a short report on the autometallograhical approaches for zinc detection in the central nervous system (CNS) by means of a modified selenium method.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.313-321
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• 1999

The purpose of these study was to investigate the use of a glass micropipette (GMP) as a vessel for vitrification of bovine IVP blastocysts, to compare the post-thaw survival rates of bovine blastocysts frozen in GMP with those frozen in OPS that have been previously investigated, and to improve the hatching rate following vitrification with GMP method. The GMP vessel permits higher freezing and warming rate than the OPS due to the higher heat conductivity of the glass and lower mass of the solution that contains the embryos. Groups of three bovine IVP blastocysts were sequentially placed into vitrification solution before being loaded into either the OPS or GMP vessels and immersed into L$N_2$within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for each 5 min, respectively, and then cultured in TCM 199 supplemented with 10% FCS for 24 h. The rate of blastocyst re-expanding did not significantly different for OPS (75.9%) and GMP (80.0%) methods (P＞0.05). The hatching rates in OPS (34.1%) and GMP (37.5%) methods were significantly lower than that in control group (54.3%) (P＞0.05). In addition, the rate of blastocyst re-expanding was significantly lower if blastocysts were vitrified in the wide portion of the micropipette rather than the narrow portion of the micropipette (83.3 vs 56.7%) (P＞0.05), even though three blastocysts were loaded per vessel. The hatching rate in 0.05% pronase solution treatment for 30, 60 and 90 see (45.9, 54.7 and 57.5%) were significantly higher than that in control (35.0%), even though there was not significantly different between 30 see and control. These results indicate that both vitrification vessels can provide high survival rates of bovine IVP blastocysts. However, the GMP vessel has the advantage over the OPS, in that the former does not need a cap to protect the vessel from floating after immersion in L$N_2$. The location of the embryos (narrow or wide portion of immersion) were considered to be limiting factors to the viability of bovine IVP embryos. The exposing in 0.05% pronase solution for 60 or 90 see can increase hatching rates of post-thaw bovine IVP blastocysts.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.199-208
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• 2000

This study was carried out to investigate of factors concerning to ultrasound-guided follicular aspiration; level of vacuum pressure, diameter of use needle, effect of FSH hormone and conception rate after embryos transfer. 1. Oocytes collection number were 4.2$\pm$2.9 e.a to luteuml phase and follicular phase were 4.4$\pm$3.5 e.a to ovaries of Hanwoo. 2. We taked proper level of aspiration vacuum pressure was 40~120 mmHg to oocytes collection. Oocytes collected number were 4.2$\pm$3.2, 4.3$\pm$3.4, 4.5$\pm$3.4 e.a. to 40, 80, 120mmHg, respectively, follicles aspiration rate were 49, 47, 45%. 3. Effect of collection needle diameter was not difference significantly(P＜0.05), oocytes collected number were 4.4$\pm$3.5 e.a to 170 and 3.0$\pm$1.8 e.a to 18G needle, collected oocytes quality were no difference significantly (P＜0.05). 4. Follicles increase number to FSH hormone injection were 6.2$\pm$2.3 e.a to intramuscle and 1.1$\pm$2.7 e.a. to epithelial injection method. 5. Conception rate derived from E.T. was 11.1% to freezing embryos and 46.2% to fresh E.T., difference significantly(P＜0.05).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.10 no.4
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• pp.189-197
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• 1977

In present study, the effects of various factors including the solvent concentration, extraction time and temperature, the ratio of sample vs extraction solvent, (w/v) and pH upon the extractability of the NaCl and alcohol soluble proteins of marine algae were investigated. Eight species of fresh algae, the major ones in consumption as food, namely Porphyra suborbiculata, Undarie pinnatifida, Hizikia fusiforme, Sargassum, fulvellum, Enteromorpha linza, Sargassum kjellmanianum, Codium coarctatum, and Ulva pertusa were used for the extraction of NaCl soluble protein and dried materials of four species, Perphyra suborbiculata, Undaria pinnatifida, Enteromorpha linza and Sargassum fulvellum were used for the extraction of alcohol soluble protein. The frozen and mascerated samples were prepared by the same method described in previous paper (Ryu, 1977). And the dried materials were moistened with alcohol solution before freezing. The effect of solvent concentration on the extractability of NaCl soluble protein differed from species. The extractability of Undaria Pinnatifide, Hizikia fusiforme, Perphyra suborbiculata, Enteromorpha linza, and Ulva pertusa reached maxima at 0.25M NaCl solution while the 1.0M for Sargassum fulvellum, Saygassum kjellmanianum and Codium coarctatum. In case of alcohol soluble proteins, it was shown at $20\%$ ethanol solution for Porphyra suborbiculata, Undaria pinnatifida, Enteromorpha linza, and Sargassum fulvellum. Variation of the ratio of sample vs solvent gave slight effect upon the extractability, but the ratio of 1:30(w/v) seemed most efficient for the extraction of NaCl soluble proteins and 100 ml solvent added to 1 g dried sample was effective in case of alcohol soluble proteins. Extraction time has a minimal effect upon the extraction of alcohol soluble protein, and approximately 21 to $43\%$ of algal protein was extracted within 1 hour. But in case of NaCl soluble protein extraction, the effect of time revealed differently from species to species resulting in that the extraction for 1 hour gave a maximum extractability in Ulva pertusa and Enteromorpha linza, 2 hours in Porphyra suborbiculata, Codium coarctatum and 3 hours in Undaria pinnatifica, Hizikia susiforme, Sargassum fulvellum and Sargassum kjellmanianum. When the NaCl soluble protein of Undaria pinnatifida and Enteromopha linza was extracted at various temperature, the most effective extraction temperature was $40^{\circ}C$ while the temperature was $50^{\circ}C$ for Undaria pinnatifida and $60^{\circ}C$ for Hixikia fusiforme, Sargassum fulvellum, Sargassum kjellmanianum and Codium coarctatum. Bus in case of alcohol soluble extraction, the optimum temperature was $30^{\circ}C$ for Enteromorpha linza and $40^{\circ}C$ for Undaria pinnatifida, Sargassum fulvellum and Porphyra suborbiculata. In the effect of pH on extractability, the maximum extractability of NaCl soluble proteins was obtained at pH 7to 8 and pH 8 to 9 for alcohol soluble protein.

• Applied Biological Chemistry
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• v.13 no.3
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• pp.207-218
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• 1970

Studies were carried out to develope the most economical and practical methods of packaging and preservation of kimchi, so commercialization of kimchi manufacture could proceed rapidly. The results obtained may be summarized as following. (1) It is generally established that the acceptable range of lactic acid content of kimchi is between 0.4% and 0.75%. Based on sensory evaluation, kimchi having lactic acid content below 0.4% and above 0.75% was not edible, and the time of optimum taste corresponded to the vicinity of 0.5% of lactic acid content. For the refrigeration storage with or without preservatives, the packaging kimchi in plastic film must be done at the lactic acid content of 0.45%, for lactic acid fermentation will continue slowly after the packaging. However, for the heat sterilized kimchi the packaging should be done at the 0.5% of lactic acid content for the best because lactic acid fermentation is completely stopped after the packaging. (2) Polyethylene, polypropylene, and polycello were chosen as suitable packaging materials. Polyethylene is cheapest among them but kimchi packaged in this film was damaged frequently in handling process and gave off kimchi flavor. On the other hand polypropylene also gave off kimchi flavor, but its higher mechanical strength gave better protection to kimchi and it had superior display effect due to the transparancy. Therefore polypropylene made much better packaging material. Polycello proved to be the best packaging material from the standpoint of physical characteristics but its price is higher than that of other plastic films. To be effective, the thickness of plastic films for packaging kimchi must exceed 0.08mm. (3) Keeping property of kimchi appeared to be excellent by means of freezing. However, by the time the frozen kimchi was thawed out at room temperature, moisture loss due to drip was extensive, rendering the kimchi too stringy. (4) Preservation of kimchi at refrigerated temperatures proved to be the best method and under the refrigerated condition the kimchi remained fresh as long as 3 months. The best results were obtained when kimchi was held at $0^{\circ}C$. (5) In general, preservatives alone were not too elective in preserving kimchi. Among them potassium sorbate appeared to be most effective with the four fold extension of self-life at $20^{\circ}C$ and two fold extension at $30^{\circ}C$. (6) In heat sterilization the thickness of packaged kimchi product had a geat effect upon the rate of heat penetration. When the thickness ranged from 1.5 to 1.8cm, the kimchi in such package could be sterilized at $65^{\circ}C$ for 20 minutes. Kimchi so heat treated could be kept at room temperature as long as one month without apparent changes in quality. (7) Among combination methods, preservation at refrigerated and heat sterilization could be favorably combined. When kimchi was stored at $4^{\circ}C$ after being sterilized at $65^{\circ}C$ for 20 minutes, it was possible to preserve the kimchi for more than 4 months.

• Food Science of Animal Resources
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• v.26 no.4
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• pp.418-425
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• 2006

This study was conducted to evaluate extraction properties of crude saponin and ginsenosides, and their effects on sensory properties of emulsified pork sausage. Non-dried ginseng root was boiled in 0 (e.g., 100% distilled water), 20, 40, 60, 80 or 100% ethanol, and powdered by a freezing dry method. Weight of dried powder for the 0% ethanol extraction was 20% of initial non-dried ginseng weight, while $20{\sim}80%$ and 100% ethanol extractions resulted in approximately 15 and 10% of their initial weights, respectively. On the other hand, crude saponin content in the dried powder was linearly increased for a higher ethanol content where 100% ethanol extraction resulted in 123.52 mg/g. LC/MS analysis of crude saponin for quantifying ginsenosides showed that Rb1, Rb2 and Rc were significantly (p<0.05) higher levels for both 80 and 100% ethanol extractions. In the case of Rg1 ginsenoside, 60, 80 and 100% ethanol extractions resulted in significantly (p<0.05) higher levels. Emulsified pork sausages containing 0, 1 or 2% ginseng extracts were smoked or non-smoked and their sensory characteristics and preference were evaluated. Smoking process significantly (p<0.05) decreased juiciness and tenderness, but the treatment significantly (p<0.05) improved flavor and consumer preference. It was particularly noticed that a 2% addition of ginseng extract prevented the adverse effects of smoking process on juiciness and tenderness while the 2% addition significantly (p<0.05) improved consumer preference. The current results implied that addition of ginseng extract in emulsified pork sausage could improve sensory quality.

• Journal of the Korean Institute of Landscape Architecture
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• v.40 no.6
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• pp.173-179
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• 2012

The experiments, which analyze the injury symptoms and diagnose growth conditions utilizing IRVT and analyzing each parts of H. helix L., had been held under a low temperature. Greenhouse and outdoor growing Genus hedera had been prepared and compared with each Genus hedera's peak and bottom leaves' surface temperature under the experimental categories $-6^{\circ}C$ and $-12^{\circ}C$. As results, analyzing the surface thermal property of peak part leaves' of outdoor growing Genus hedera, at experimental categories $-6^{\circ}C$, $-12^{\circ}C$ were ranged from $-2^{\circ}C{\sim}-7^{\circ}C$ and $-2^{\circ}C{\sim}-15^{\circ}C$. On the other hand, the surface thermal property of bottom part leaves at experimental categories $-6^{\circ}C$, $-12^{\circ}C$ were ranged $-2^{\circ}C{\sim}-11^{\circ}C$ and $-1^{\circ}C{\sim}-12^{\circ}C$. It appears that the thermal properties of leaves' surface on $-6^{\circ}C$ peaks and $-12^{\circ}C$ bottoms were more broadband than bottoms and peaks. It means that the peaks were more sensitive than bottoms, as like $-2^{\circ}C{\sim}-15^{\circ}C$, $-1{\sim}-12^{\circ}C$. Moreover, as similar results had seen to leaves surface temperature added to cold wind conditions. How the cold wind damaged the outdoor growing Genus hedera, analyzed the surface thermal property by IRVT data under $0^{\circ}C$, $-2^{\circ}C$, $-4^{\circ}C$ condition, it resulted to $-6.2^{\circ}C$, $-6.8^{\circ}C$, $-7.5^{\circ}C$. It appeared more $3.5{\sim}6.2^{\circ}C$ low temperature than experimental setting point. In addition, each parts thurmal property of peaks and bottoms was not similar, it referred to each parts' sensitivities of low temperature were different on the peak and bottom leaves surface temperature.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.117-125
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• 2001

Vitrification of oocytes has been applied recently fur pigs, but remains elusive. The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) on in vitro survival of porcine oocytes. When immature follicular oocytes frozen-thawed were cultured for in vitro maturation, maturation rates to metaphase-II stage were higher in oocytes with (25%) than without (15%) cumulus cells. After In vitro fertilization of oocytes frozen-thawed, the maturation rates were also significantly (P<0.05) higher in oocytes with (41%) that than without (17%) cumulus cells. However, the penetration rates were higher in oocytes without (19%) that than with (9%) cumulus. In another experiment, porcine oocytes matured in vitro were frozen and thawed for in vitro fertilization. The penetration rates were higher than in oocytes without (35%) that than with (26%) cumulus cells. However, the proportions of oocytes dead after in vitro fertilization were significantly (P<0.05) higher in oocytes with that than without cumulus cells. On the other hand, the rates of penetration and dead oocytes at 6 h after in vitro fertilization were not significant differences between oocytes with and without cumulus cells. However, the proportions of dead oocytes with (18%) and without (16%) cumulus cells were higher than in oocytes of control group (0%). These finding indicated the possible broader application for OPS, as they demonstrated that the maturation and fertilization in vitro by frozen-thawed oocytes may be accompained by cumulus cells and culture periods according to the requirements of the survival ability after freezing of mature and immature oocytes in pigs.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.255-262
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• 2006

The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.231-238
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• 1999

The objective of this study was to examine the effect of developmental stage and embryo age of in vitro produced bovine blastocysts after vitrification and thawing. In vitro cultured day 8 blastocysts after IVF were equilibrated 20% ethylene glycol (EG) for 3 min. and were vitrified using EFS40, which is consisted of 40% EG, 18% ficoll, 0.3M sucrose and 10% FBS added in mDPBS for 30 sec. before being plunged into $LN_2$. Also, survival in vitro was assessed by re-expansion and hatching or hatched at 24 hand 48 h postwarming, respectively. The results obtained in these experiments were summarized as follows; 1) When the embryos were cultured for 8 day after IVF, 41.0% of the cleaved embryos developed to the blastocysts (early; 7.6%, expanded; 22.9%, hatching; 4.6% and hatched; 5.9%). 2) When the embryos were exposed or vitrified to the freezing solution, the re-expansion of vitrified embryos (73.3%) was significantly lower than that of control and exposed embryos (100, 97.0%) (p<0.05). But the formation rate of hatching or hatched blastocysts of vitrified embryos (66.7, 46.7%) at 48h after thawing was similar to that of exposed embryos (66.7, 39.4%) but not control (100, 100%) (p<0.01). However, in the total cell numbers of those developed hatched blastocysts, there were not significantly different among the treatment groups. 3) When the embryo survival rates by different developmental stage were examined, the re-expansion was not different among the groups $(64.5{\sim}75.6%)$. After warming 48 h, the hatching and hatched formation of early blastocysts (25.8, 9.7%) was significantly lower than those of expanded (69.7, 39.4%) and hatching blastocysts (53.3, 43.3%) (p<0.05). 4) In addition, when the expanded blastocysts at day 7, 8 and 9 were vitrified, the re-expansion of day 8 and 9 embryos was significantly lower than that of day 7 (day 7; 93.9%, day 8; 75.8% and day 9; 87.5%) (p<0.05). However, the rates of development to hatched blastocysts were no difference among the groups (day 7; 36.4%, day 8; 36.4% and day 9; 31.3%). These results suggested that in vitro produced expanded or hatching blastocysts can be efficiently cryopreserved by the two-step vitrification method using EFS40.