• 한국균학회지
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• 제31권3호
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• pp.187-191
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• 2003

뽕나무 버섯균은 식용버섯으로써 노인병, 중풍, 현기증, 두통, 신경쇠약, 불면증, 사지마비, 소아마비, 간질 등에 효과가 있는 것으로 알려져 있으므로, 뽕나무 버섯 균사체의 고농도 배양을 위한 최적 조건확립 하고자 하였다. 뽕나무 버섯균사체 생산의 최적배지 선정을 위하여 7종의 배지를 대상으로 수행한 결과 맥아즙배지가 뽕나무 버섯 균사체 생산에 가장 우수하였다. 맥아즙 배지에서 뽕나무 버섯균 균사체 생산을 위한 적정조건은 맥아즙 농도 $15\;Brix^{\circ}$이었으며 온도, pH, 배양기간의 조건을 반응표면 분석법에 의해서 실행한 결과, $25.9^{\circ}C$, pH 5.72, 15.22일의 배양에서 3,40 g/50 ml의 최대 예측치를 나타내었다. 이는 실제 실험결과치와도 대체로 일치하는 경향이었다.

• 한국균학회지
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• 제31권3호
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• pp.168-174
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• 2003

C. militaris 균사를 콜로이달 키틴이 첨가된 액체 배지에서 배양한 후 황산암모늄 분별침전, 이온 교환 및 겔 여과 크로마토그래피를 이용하여 배양액 중의 chitianse를 분리 정제하였다. 이 효소의 최적 pH와 온도는 각각 5.5와 $35^{\circ}C$이었으며 겉보기 분자량은 48.5 kDa이었고, 그 Km 값은 0.57 mM이었다. 이 효소는 $Cu^{2+},\;Mn^{2+},\;Hg^{2+},\;Zn^{2+},\;CO_{3}^{2-},\;SO_4^{2-},\;CN^-$ 및 OCN^-$ 이온에 의하여 활성이 억제되었으나, $Mg^{2+}$ 및 $K^+$ 이온에 의하여 활성이 약간 증가되었다. 또한 효소 단백질의 아미노산 잔기와 선택적으로 반응하는 무수말레인산, 무수아세트산 및 N-bromo succinimide에 의하여 84.0% 활성이 억제되어 카르복실기를 가진 아미노산 잔기가 이 효소의 활성 부위에 중요한 역할을 함을 알았다. NAG6에 의한 기질 분해 특이성 실험을 통하여 이 효소는 endo-형태의 chitinase임을 알았다.

• Preventive Nutrition and Food Science
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• 제3권1호
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• pp.85-91
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• 1998

To investigate the effects of antioxidative vitamins in combination with various fatty acids on breast cancer cell proliferation, MDA-MB231 human breast cancer cells were cultured for 3 days in the serum-free Iscove's modified Dulbecco's medium (IMDM) supplemented with 1.25mg/ml delipidized bovine serum albumin and 10㎍/ml insulin. Alpha-tocopherol, ascorbic acid or both vitamins were added to the medium at the concentrations of 10 and 50μM in the presence of 3μg/ml of oletic(Oa), linoleic(LA) α-linoleinic(LNA) and docosahexaenoic acid(DHA). Cell growth was reduced significantly by α-tocopherol in a dose-dependent manner, but not affected by ascorbic aicd. The four different fatty acids did not have significant effects on cell growth, although DHA exerted inhibitory effect on the growth after 1 day. However, the each fatty acid was well incorporated into celluar lipid as such or elongated forms. Addition of α-tocopherol remarkably increased its celluar contents and reduced cellular levels of thiobarbituric acid substances (TBARS) that were elevated notably in the presence of DHA in the culture media. But ascorbic acid addition did not change much of either cellular α-tocopherol or TBARS contents. northern blot hybridization showed that tumor supressor gene ρ53 was most highly expressed by the combination of ρ-tocopherol and DHA in 8 hours of cell culture. In conclusion , the growth inhibitory effect of vitamin E suggests that breast cancer cell proliferation is reduced by the mechanism other than cytotoxicity of lipid peroxide and it is related to expressionof tumor supprosser gene p53, that can be increased by both vitamin E and n-3 fatty acid, DHA.

• KSBB Journal
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• 제21권2호
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• pp.123-128
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• 2006

황기의 형질전환된 모상근으로부터 약리물질인 astragalosides I, II, 그리고 III을 효율적으로 생산하기 위하여 다양한 물리화학적 조건들 즉, 배지, 초기 당농도, 교반속도, 초기 접종농도, 광조사, 그리고 chitosan 처리에 한 elicitation 효과를 조사 하였다. 선발된 AG-04 clone의 성장과 astragalosides의 함량은 4종의 배지들 중 SH배지에서 가장 좋았으며, 배지에 첨가하는 초기 당 농도는 3%와 6%에서 각기 건물중과 astragalosides의 함량이 가장 높게 나타났다. 또한, 초기 접종량은 50 ml 배지에 500 mg(FRW)씩을 주입하는게 가장 효과적이었으며, 18시간 광조사가 되는 Shaking incubator에서 90 rpm과 120으로 교반시켰을 때 모상근의 성장과 astargalosides의 함량이 높게 나타났다. 배양 2주 후 chitosan의 처리는 모상근의 성장에는 크게 영향을 미치지 않았으나, 100 mg/L 처리구에서 대조구의 약 2.1배의 astragalosides 함량 증가를 가져왔다. 이와같은 연구결과는 황기의 대량배양을 통한 saponins 생산 연구에 효율적으로 활용될 수 있을 것으로 본다.

• 한국수정란이식학회지
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• 제31권1호
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• pp.39-46
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• 2016

Cryopreservation has been applied successfully in many mammalian species. Nevertheless, pig embryos, because of their greater susceptibility to cryoinjuries, have shown a reduced developmental competence. The aim of this study was to evaluate the survival status of vitrified-warmed porcine embryos. Forced blastocoele collapse (FBC) and non-FBC blastocysts are vitrified and concomitantly cultured in culture media which were supplemented with/without fetal bovine serum (FBS). Porcine vitrified-warmed embryos were examined in four different methods: group A, non-FBC without FBS; group B, non-FBC with FBS; group C, FBC without FBS; group D, FBC with FBS. After culture, differences in survival rates of blastocysts derived from vitrified-warmed porcine embryos were found in group A~D (39.5 (A) vs 52.5 (B) and 54.8 (C) vs 66.7% (D), respectively, p<0.05). Reactive oxygen species (ROS) level of survived blastocysts was lower in group D than that of another groups (p<0.05). Moreover, total cell number of survived blastocysts was higher in group D than that of other groups (p<0.05). Otherwise, group D showed significantly lower number of apoptotic cells than other groups ($2.0{\pm}1.5$ vs $3.2{\pm}2.1$, $2.8{\pm}1.9$, and $2.7{\pm}1.6$, respectively, p<0.05). Taken together, these results showed that FBS/FBC improves the developmental competence of vitrified porcine embryos by modulating intracellular levels of ROS and the apoptotic index during the vitrification/warming procedure. Therefore, we suggest that FBS and FBC are effective treatment techniques during the vitrification/warming procedures of porcine blastocysts.

• 한국수정란이식학회지
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• 제28권4호
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• pp.337-342
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• 2013

Salmonellosis is one of the life-threating diseases of goat in Bangladesh. Therefore, the present study was designed to study the prevalence of Salmonellosis, and isolation and characterizations of the Salmonella spp. from apparently healthy and diarrheic goat. A total of 47 faces samples were collected from selected place and cultured onto different prescribed medium to isolate it. In this study, 12.76% (6/47) samples were found to be positive for Salmonella spp. During culture on SS agar medium, all of the Salmonella isolates produced round, smooth, opaque, translucent and black color colonies on SS agar media. All of the isolated Salmonella spp. fermented dextrose, maltose and mannitol with production of acid and gas but did not ferment sucrose and lactose. However, these isolates had showed Indole and Voges-Proskauer test negative, Methyl-Red test positive. All of these isolates were subjected to rapid plate agglutination test with polyvalent "O" (Poly 'O') and polyvalent "H" (poly 'H') antisera where positive agglutination were observed. They were highly sensitive to ciprofloxacin, spiramycin and gentamycin; moderately sensitive to oxytetracyline, streptomycin and amoxicillin; less sensitive to sulphamethoxazole and resistant to penicillin-G. Based on the present findings, it may be concluded that the investigated Salmonella spp. from goats might be S. typhimurium, S. enteritidis, S. brandenburg, S. salford, S. newbrunswick, S. newport or S. dublin. Further study will be needed, therefore it requires further characterization using other serological and molecular techniques.

• Archives of Plastic Surgery
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• 제34권2호
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• pp.141-148
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• 2007

Purpose: The object of this study was to evaluate the development of continuous osteogenic differentiation and bone formation after the subcutaneous implantation of the tissue-engineered bone, in vitro. Methods: Human adipose-derived stem cells were obtained by proteolytic digestion of liposuction aspirates. Adipose-derived stem cells were seeded in PLGA scaffolds after being labeled with PKH26 and cultured in osteogenic differentiation media for 1 month. The PLGA scaffolds with osteogenic stimulated adipose-derived stem cells were implanted in subcutaneous layer of four nude mice. Osteogenesis was assessed by RT-PCR for mRNA of osteopontin and bone sialoprotein(BSP), and immunohistochemistry for osteocalcin, and von Kossa staining for calcification of extracellular matrix at 1 and 2 months. Results: Implanted PLGA scaffold with adipose-derived stem cells were well vascularized, and PLGA scaffolds degraded and were substituted by host tissues. The mRNA of osteopontin and BSP was detected by RT-PCR in both osteogenic stimulation group and also osteocalcin was detected by immunohistochemistry at osteogenic stimulation 1 and 2 months, but no calcified extracellular deposit in von Kossa stain was found in all groups. Conclusion: In vivo, it could also maintain the characteristics of osteogenic differentiation that adipose-derived stem cells within PLGA scaffold after stimulation of osteogenic differentiation in vitro, but there were not normal bone formation in subcutaneous area. Another important factor to consider is in vivo, heterologous environment would have negative effect on bone formation as.[p1]

• 한국임상수의학회지
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• 제20권4호
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• pp.496-500
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• 2003

A one-year-old male Japanese Chin with anorexia, retching, dyspnea and continuous coughing was brought to the Veterinary Teaching Hospital, Chungbuk National University. Chest radiographs showed moderate regional alveolar pattern with mild interstitial patterns in the caudo-dorsal lung fields and the ill-defined mass in the perihilar area which is consistent with perihilar lymphadenopathy. Although the dog showed severe eosinophilia in the complete blood count, the serum profile values were within normal ranges. There was no indication of any parasite infestation in the direct and floatation examination of feces, skin scraping test and heartworm examination. There was no growth of bacteria and fungi in the selected media such as Mueller Hinton broth, Sabouraud Dextrose agar and Potato Dextorse agar, which were inoculated with tracheal fluid collected using endotracheal tube and cultured for 3 days. In the tracheal fluid smear, most prominent cells were eosinophils, which are a almost 80% of total cells and other cells such as leukocytes, neutrophils and ciliated colummar cells were also observed. Any parasite was also not detected in its smear. Prednisolone (PDS; 1 mg/kg, BID SC), aminophylline (10 mg/kg, TID IV) and nebulization with gentamicin (50 mg) plus saline (3 ml) were given for 1 week. At 3rd day of treatment, blood eosinophil value was return to normal range and pulmonary condition was also improved. The allergen test with serum performed during therapy was positive in the 19 index including milk, barley, tomato pomace, catfish, bonito, house dust and wool, and borderline in 10 index including wheat, house dust mites and house fly. The patient is responding well to PDS therapy. Based on these findings, a possible diagnosis of pulmonary infiltration with eosinophils was made in this dog.

• Toxicological Research
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• 제12권1호
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• pp.69-79
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• 1996

Primary culture of cerebellar neuronal cells derived from 8-day old Long-Evans rats was used. Pure granule cells, astrocytes or mixed cells culture systems were prepared. These cells were differentiated and developed synaptic connections. And the astrocytes were identified by immunostaining with glial fibrillary acidic protein (GFAP). Methamphetamine (MAP), which acts on dopaminergic system and cadmium (Cd), a toxic heavy metal, were applied and biochemical assays and electrophysiological studies were performed. $LC_50$ values estimated by MTT assay of MAP and Cd were 3 mM and 2$\mu M$ respectively. Cells were treated with 1 mM or 2 mM MAP and 1$\mu M$ $CdCl_2$ for 48 hour, and the incubation media were analyzed for the content of released LDH. MAP (2 mM) and Cd significantly increased the LDH release. Cell viability was decreased in both groups and some cytopathological changes like cell swelling or vacuolization were seen. The cerebellar granule cells were used for measuring membrane currents using whole-cell clamp technique. Sodium and potassium currents were not affected by MAP neither Cd, but calcium current was significantly reduced by Cd but not affected by MAP. Therefore, in vitro neurotoxicity test system using neuronaI cells and astrocytes cultures were established and can be used in screening of potential neurotoxic chemicals.

• Parasites, Hosts and Diseases
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• 제31권4호
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• pp.379-381
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• 1993

우태아혈청의 성분이 Toxoplasma의 숙주세포 침투를 억제하는 현상을 보여 이를 보고하고자 한다. MDCK세포를 숙주로 하여 Toxoplasma의 RH주를 첨가하였을 때, 우태아혈청 농도 1%(v/v) 에서 억제 효과가 나타나기 시작하여 5%일때 침투능을 반감시켰다. 우태아절청의 비동화 여부는 억제 효과에 영향을 미치지 못하였으며, 우태아혈청을 $95^{\circ}C$에서 10분간 처리하여 억제물질의 기능을 파괴시킬 때 억제 효과는 약간 감소되었다. Toxoprasma를 먼저 우태아혈청으로 처리한 후 숙주 세포에 첨가하였을 때 침투능에는 효과를 미치지 못하였으나, Tomplasma를 숙주세포와 배양하면서 우태아혈청을 첨가하였을 때에는 시간의 경과에 따라 침투능을 억제시켰다. 이상의 결과로 우태아혈청에 Toxoplasma의 숙주세포 침투를 억제하는 성분이 존재하는 것을 확인하였으며, 억제 방법이 침투하는 순간에 일어나며, 억제물질의 기능을 통해서라기 보다는 구조를 통해서 이루어진다고 추정할 수 있었다.