• Journal of Chest Surgery
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• v.36 no.4
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• pp.219-228
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• 2003

Background:Liquid nitrogen freezing techniques have already met with widespread success in biology and medicine as a means of long-term storage for cells and tissues. The use of cryoprotectants such as glycerol and dimethylsulphoxide to prevent ice crystal formation, with carefully controlled rates of freezing and thawing, allows both structure and viability to be retained almost indefinitely. Cryopreservation of various tissues has various con-trolled rates of freezing. Material and Method: To find the optimal freezing curve and the chamber temperature, we approached the thermodynamic calculation of tissues in two ways. One is the direct calculation method. We should know the thermophysical characteristics of all components, latent heat of fusion, area, density and volume, etc. This kind of calculation is so sophisticated and some variables may not be determined. The other is the indirect calculation method. We performed the tissue freezing with already used freezing curve and we observed the actual freezing curve of that tissue. And we modified the freezing curve with several steps of calculation, polynomial regression analysis, time constant calculation, thermal response calculation and inverse calculation of chamber temperature. Result: We applied that freezing program on mesenchymal stem cell, chondrocyte, and osteoblast. The tissue temperature decreased according to the ideal freezing curve without temperature rising. We did not find any differences in survival. The reason is postulated to be that freezing material is too small and contains cellular components. We expect the significant difference in cellular viability if the freezing curve is applied on a large scale of tissues. Conclusion: This program would be helpful in finding the chamber temperature for the ideal freezing curie easily.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.541-550
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• 1996

Tetracycline antibiotics have been widely used not only therapeutics but feed additives. There are many methods for the isolation and determination of tetracycline antibiotics in animal muscle tissue. But those methods take much time and labor, so it is difficult to analyse many samples simultaneously. A rapid isolation method and liquid chromatographic determination of tetracycline antibiotics in animal muscle tissue (bovine, porcine, chicken) is presented. Blank control and tetracyclines fortified samples (0.5g) were blended with $C_{18}$ containing 0.05g each of oxalic acid and disodium ethylenediaminetetraacetate. After homogenize, homogenate was transferred to glass column made from 10ml glass syringe and compressed to 4~4.5ml volume. A column made from the $C_{18}$/meat matrix was washed with hexane (8ml) and dichloromethane (8ml, if needed), following which the tetracyclines were eluted,vith methanol or 0.01M methanolic oxalic acid (8ml). The eluates containing tetracyclines analytes were free from interfering compounds when analysed by HPLC with UV detection (photodiode array at 360nm). Standard curve for each tetracycline showed a linear response at the range of $0.05{\sim}1.0{\mu}g/ml$ and tetracycline antibiotics were eluted within 4ml of eluted volume. All tetracycline antibiotics except tetracycline were stable during the concentration process at $40^{\circ}C$ and time required for concentration was 3~4 hours. Fortified samples containing oxalic aicd and EDTA represented more good recoveries than those of not-contained sample. Recoveries were 91.8~110.1% (oxytetracycline; OTC), 57.7~79.5% (tetracycline; TC), 78.1~88.6% (chlortetracyclines; CTC) and 88.4~100.6% (doxycycline; DC) in pork tissue, 101.1~126.8% (OTC), 66.4~75.4% (TC), 79.2~88.1% (CTC) and 69.3~86.7% (DC) in beef tissue, and 90.8~95.6% (OTC), 66.2~84.4% (TC), 75.7~77.2% (CTC) and 55.6~80.7% (DC) in chicken muscle tissue. The detection limits validated in muscle tissue by this method were $0.05{\mu}g/g$ for OTC and TC, and $0.1{\mu}g/g$ for CTC and DC.

• Korean Journal of Medicinal Crop Science
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• v.26 no.3
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• pp.220-226
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• 2018

Background: Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz. belong to the family (Asteraceae). Their rhizomes, called white Atractylodes rhizomes, are used in traditional medicine. To address some issues with their cultivation, we recently developed eight hybrid cultivars by interspecific hybridization of A. japonica and A. macrocephala. This study was conducted to screen the hybrid cultivars that have high amounts of active ingredients and yield ability. Methods and Results: Experiments were conducted using the eight hybrid cultivars and A. macrocephala (control cultivar) in the experimental field of the department of Herbal Crop Research located in Eumseong, South Korea. We investigated the growth characteristics of the aerial and underground parts. Among the cultivars, 'Sanwon' had the highest rhizome dry weight (53.8 g/plant), followed by 'Dachul' (50.0 g/plant). In addition, the content of atractylenolide I, II, III and total active ingredients were investigated using high-performance liquid chromatography. Compared with A. macrocephala, most of the inter-specific hybrid cultivars had a higher content of active ingredients and yield ability. Conclusions: Through study, we established the superior quality of Atractylodes inter-specific hybrid cultivars. In particularly, it was found that 'Dachul' may be grown as a superior cultivar, with high amount of active ingredients as well as yield ability.

• Korean Journal of Medicinal Crop Science
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• v.26 no.3
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• pp.227-232
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• 2018

Background: Angelica gigas Nakai is a perennial herb belonging to the family Umbelliferae. Its roots are utilized in traditional medicine. The aim of this study was to increase the yield of and the content of two indicator components (decursin and decursinol angelate) in A. gigas Nakai. Methods and Results: The roots of A. gigas Nakai were harvested 4-times from late August to late October in 2017. Two agents (trace element-TE, and plant hormone-HM) were applied 4 times at intervals of 2 weeks. The content of the two indicator components were analyzed by high performance liquid chromatography. The HM treatment showed the greatest increase in underground part yield and root diameter. The content of the two indicator components in the control (non-treatment) group was the highest in the underground part, but was higher in the aerial parts in the agent treatment group. After treatment with the agents, the content of the indicator components tended to decrease in the underground part. However, the total content of the indicator components in the two agent treatment groups exceeded the level of 6% set by the Korean Pharmacopoeia. Conclusions: The highest underground part yield was found in the HM treatment group, while the highest content of decursin and decursinol angelate were found in the control group. This study provides basic information for yield improvement in A. gigas.

• Journal of Pharmaceutical Investigation
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• v.40 no.2
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• pp.125-131
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• 2010

A bioequivalence study of LANIDIEM$^{(R)}$ tablet 4 mg (Samil. Co., Ltd.) to Vaxar$^{(R)}$ tablet 4 mg (GlaxoSmithKline Co., Ltd.) was conducted according to the guidelines of Korea Food and Drug Administration (KFDA). Forty healthy male Korean volunteers were enrolled in the study and thirty six volunteers completed the study according to the protocol. Thirty six volunteers received each medicine at the lacidipine dose of 4 mg in a $2{\times}2$ crossover study. There was one week wash-out period between the doses. Plasma concentrations of lacidipine were monitored by a high performance liquid chromatography - tandem mass spectrometry (LC-MS/MS) for over a period of 24 hours after drug administration. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 24 hr) was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_t$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the crossover design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for LANIDIEM$^{(R)}$/Vaxar$^{(R)}$ were log 0.8102~log 1.0417 and log 0.8493~log 1.1439, respectively. These values were within the acceptable bioequivalence intervals of log 0.80~log 1.25. Thus, our study demonstrated the bioequivalence of LANIDIEM$^{(R)}$ tablet 4 mg and Vaxar$^{(R)}$ tablet 4 mg with respect to the rate and extent of absorption.

• Analytical Science and Technology
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• v.18 no.3
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• pp.250-256
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• 2005

A selective method of high performance liquid chromatography with UV detector has been applied to determine 4 tetracycline antibiotics in the animal food, simultaneously. The targets were chlortetracycline (CTC), doxycycline (DC), oxytetracycline (OTC), and tetracycline (TC) that are used routinely in veterinary medicine for prevention and control of disease. Food samples were beef, pork, chicken, milk, whole egg, flatfish (Limanda yokohamae), jacopever (Sebastes hubbsi), seabream (Chrysophrys major), eel (Anguilla japonica) and lobster (Hommarus americanus). After extracting food samples with 20% trichloroacetic acid and McIlvaine buffer, they were purified by a $C_18$ SPE cartridge with 0.01M methanolic oxalic acid solution. The concentrated residue was re-dissolved in methanol, filtered, cleaned up and analyzed on a $C_18$ column. The mobile phase was a mixture of 0.01M oxalic acid and acetonitrile with a gradient ratio from 85:15 to 60:40. The UV wavelength was 365 nm. The overall recoveries were ranged from 71% to 98% and the limit of detections were 0.022 for CTC, 0.012 for DC and OTC and 0.009 mg/kg for TC at signal/noise > 3, respectively. As results, CTC, DC and TC were not detected in all selected food samples, however, OTC was detected in meat and fishes. The determined level of OTC was 0.04 ppm for pork, 0.17 ppm for flatfish and 0.05 and 0.08 ppm for jacopever, that were within the Maximum Residue Limits (MRLs) in the food.

• Journal of Periodontal and Implant Science
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• v.24 no.1
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• pp.144-154
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• 1994

Final goal of periodontal treatment is to reconstruct the destructed periodontal tissue as well as to remove the necrotic pathologic elements. The purpose of this study is to investigate on the effect of Zizyphi extract to the inhibitory ability on collagenolytic activity of P gingivalis, biologic activity of gingival fibroblasts, and on the collagen and protein synthesis of gingival fibroblasts. Gingival fibroblast from giniva of first bicuspids from patient for orthodontic treatment were used and cultured. For the measurement of inhibitory ability of collagenolytic activity, crude enzyme was extracted and used on the basis of modified Ono's method. On the inhibition of collagenolytic enzyme from herbal extracts, collagenokit CLN-100 were used. The cellular activity of gingival fibroblast, were studied using MTT solution and measured optical density on 570mm by ELISA reader. To measure the effects on the ability of whole protein and collagen synthesis, cell membrane was destructed with ultrasonic grinder after culturing, centrifuged and counted by liquid scintilation counter. The inhibitory effects on producing of $IL-l{\beta}$ by monocyte, after promotion of producing $IL-l{\beta}$ by LPS, were compared with the mixture of herbal extracts and other drugs using thymocyte stimulation assay. About inhibitory effects of $PGF_2$. by gingival fibroblasts, herbal extract was compared with the addition of the other control groups using enzyme imunoassay. On the inhibition of collagenolytic activity by P. gingivalis, benzene extracts showed the most efficient inhibitory effects among the $19{\mu}g/ml$ of the compared extracts and 40.5% by Tetracycline. On the cellular activity promoting effects, compared extracts showed a bit of more effects than PDGF of $100{\mu}g/ml$ concentration and IGF of $20{\mu}g/ml$ concentration. All of the PDGF, IGF, Zizyphi Fructus extract should increase in collagen synthesis, but especially 70% ethylalcohol extracts of Zizyphi Fructus showed comparably high effects among the compared extracts. Effects on whole protein synthesis were slightly increased on every extract but especially 70% ethylalcohol extract showed significantly effective than any other estract. On the inhibitory effects of Zizyphi Fructus $IL-l{\beta}$ production by monocyte, compared extracts showed 70% of highly inhibitory effect than that of 60% inhibition effects on controlled group and each extracts showed no significant difference. In $PGF_2$ production inhibitroy effect of Zizyphi Fructus gingival fibroblasts, Herbal extracts showed 70% of inhibition comparing with tat of 90.2% of controlled group, but each extracts showed similar effects excluding the $H_2O$ extracts. These results suggested that Zizyphi Fructus might be useful medicine for inhibition of inflammatory mediator including $IL-l{\beta}$ and $PGF_2$.

• Korean Journal of Plant Resources
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• v.22 no.5
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• pp.454-460
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• 2009

Phyllanthus urinaria was an important species in Korea and distributed in all around of Korea. The roots and stems of this plant have been used for natural medicine for the treatment of diabetes, the hepatitis B virus and disturbances of the kidney and urinary bladder. Production of adventitious roots in P. urinaria by in vitro cultures could be used as alternatives materials. Shoot and root segments from P. urinaria seedling were cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg/L IBA and 30 g/L sucrose. After 4 weeks of culture, the highest induction of adventitious roots was obtained from the shoot part. Frequency of adventitious root formation on medium with various kinds of auxins (IAA, NAA, 2,4-D, and IBA) and various concentrations of IBA (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L) was tested. The maximun induction of adventitious root was obtained on medium with 0.5 mg/L IBA. In liquid culture, growth of root was best on medium supplemented with 30 g/L sucrose. Adventitious roots were cultured in 5 L bioreactor containing 1/2 MS medium supplemented with 0.5 mg/L IBA and 30 g/L sucrose and mass-production of adventitious roots was successfully achieved. These results revealed the first attempt for the production of adventitious roots in P. urinaria.

• Journal of Microbiology
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• v.45 no.5
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• pp.460-465
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• 2007

This study was carried out in order to investigate the potential of using plant oils derived from Leptospermum petersonii Bailey and Syzygium aromaticum L. Merr. Et Perry as natural antifungal agents. The antifungal effects of essential oils at concentrations of 0.05, 0.1, 0.15, and 0.2 mg/ml on the dermatophytes Microsporum canis (KCTC 6591), Trichophyton mentagrophytes (KCTC 6077), Trichophyton rubrum (KCCM 60443), Epidermophyton floccosum (KCCM 11667), and Microsporum gypseum were evaluated using the agar diffusion method. The major constituents of the active fraction against the dermatophytes were identified by gas chromatography-mass spectrometry and high-performance liquid chromatography analysis. The antifungal activities of S. aromaticum oil (clove oil) against the dermatophytes tested were highest at a concentration of 0.2mg/ml, with an effectiveness of more than 60%. Hyphal growth was completely inhibited in T. mentagrophytes, T. rubrum, and M. gypseum by treatment with clove oil at a concentration of 0.2 mg/ml. Eugenol was the most effective antifungal constituent of clove oil against the dermatophytes T. mentagrophytes and M. canis. Morphological changes in the hyphae of T. mentagrophytes, such as damage to the cell wall and cell membrane and the expansion of the endoplasmic reticulum, after treatment with 0.11 mg/ml eugenol were observed by transmission electron microscopy (TEM). At a concentration of 0.2 mg/ml, L. petersonii oil (LPO) was more than 90% effective against all of the dermatophytes tested, with the exception of T. rubrum. Geranial was determined to be the most active antifungal constituent of L. petersonii oil. Taken together, the results of this study demonstrate that clove and tea tree oils exhibited significant antifungal activities against the dermatophytes tested in this study.

• Korean Journal of Medicinal Crop Science
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• v.7 no.1
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• pp.45-50
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• 1999

Timosaponin A III, an active and major compound, was isolated from rhizomes of Anemarrhena asphodeloides. The quantitative analysis of timosaponin AIII was performed by a high performance liquid chromatographic(HPLC) method using ELSD and the useful extraction method for HPLC analysis was examined as well. This HPLC method can be utilized as the standard analytical method for the evaluation of the quality of Anemarrhena rhizoma in the steps of breeding and cultivation. Additionally, the HPLC analysis method can be useful for the evaluation of the quality of Anemarrhena rhizoma sold as a traditional medicine in current markets.