• 한국응용과학기술학회지
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• 제34권3호
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• pp.673-682
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• 2017

식물 및 동물성 단백질 유래 펩타이드 형태의 단백질 가수 분해물들은 항산화, 고혈압 완화, 면역조절, 진통완화 및 항균작용 등 생리활성이 있는 것으로 알려져 왔다. 본 연구는 연산오계란 단백질 가수 분해물을 Ultrafiltration를 이용하여 HDS(분획되지 않은 가수 분해물), 1 kDa, 5 kDa, 10 kDa, 50 kDa로 분획된 기능성 펩타이드의 DPPH radical scavenging activity, superoxide anion radical scavenging activity, hydroxyl radical scavenging activity 및 $Fe^{2+}$ chelation ability을 평가하였다. 그 결과 DPPH radical scavenging activity 최대값은 1 kDa(70.83 %), hydroxyl radical scavenging activity 최대값은 5 kDa (47.01 %), superoxide anion radical scavenging activity 최대값은 5 kDa(40.57 %), $Fe^{2+}$ chelation ability 최대값은 5 kDa(29.87 %)로 나타났다. Ultrafiltration를 이용하여 fractionation된 단백질 가수 분해물의 항산화 저해 능력 $IC_{50}$ 평가하였다. 그 결과 HDS의 최대값은 superoxide anion radical scavenging activity($IC_{50}$, 5.42 mg/ml)이고, 1 kDa의 최대값은 $Fe^{2+}$ chelation ability($IC_{50}$, 1.67 mg/ml)이고, 5 kDa의 최대값은 $Fe^{2+}$ chelation ability($IC_{50}$, 2.09 mg/ml)이고, 10 kDa의 최대값은 $Fe^{2+}$ chelation ability($IC_{50}$, 2.61 mg/ml)이고, 50 kDa의 최대값은 $Fe^{2+}$ chelation ability($IC_{50}$, 4.53 mg/ml)이다. 그러므로 본 연구 결과를 바탕으로 5 kDa를 이용하여 오계란 단백질에서 분획한 펩타이드는 항산화 기능성 식품소재로서 활용할 가치가 높을 것으로 기대한다.

• Parasites, Hosts and Diseases
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• 제52권6호
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• pp.613-619
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• 2014

Neospora caninum (Apicomplexa; Sarcocystidae) is a protozoan that causes abortion in cattle, horses, sheep, and dogs as well as neurological and dermatological diseases in dogs. In the central nervous system of dogs infected with N. caninum, cysts were detected that exhibited gliosis and meningitis. Flavonoids are polyphenolic compounds that exhibit antibacterial, antiparasitic, antifungal, and antiviral properties. In this study, we investigated the effects of flavonoids in a well-established in vitro model of N. caninum infection in glial cell cultures. Glial cells were treated individually with 10 different flavonoids, and a subset of cultures was also infected with the NC-1 strain of N. caninum. All of the flavonoids tested induced an increase in the metabolism of glial cells and many of them increased nitrite levels in cultures infected with NC-1 compared to controls and uninfected cultures. Among the flavonoids tested, 3',4'-dihydroxyflavone, 3',4',5,7-tetrahydroxyflavone (luteolin), and 3,3',4',5,6-pentahydroxyflavone (quercetin), also inhibited parasitophorous vacuole formation. Taken together, our findings show that flavonoids modulate glial cell responses, increase NO secretion, and interfere with N. caninum infection and proliferation.

• Clinical and Experimental Reproductive Medicine
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• 제25권2호
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• pp.161-170
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• 1998

Protein expression patterns of matrix metalloproteinases (MMPs) were examined in mouse reproductive organs during estrous cycle. Estrous cycle was classified into diestrus, proestrus, estrus or metestus and MMP expression was analyzed by zymography using gelatin as a substrate. Uterine fluid (UF) obtained both at diestrus and proestrus exhibited 4 major MMPs including 106kDa, 64kDa, 62kDa and 59kDa gelatinases. However, in UF at estrus, the gelatinolytic activity of 64kDa MMP disappeared and that of 106kDa and 62kDa MMPs dramatically decreased. At metestrus, 64kDa MMP activity reappeared and 106kDa and 62kDa MMP exhibited increased activities such that the band intensity of 106kDa was comparable to that in UF at diestrus. Gelatinolytic activity of 59kDa MMP was not changed throughout the cycle. Both ovarian and oviductal tissue homogenate revealed 4 MMPs which corresponded to the 4 MMPs of UF. However, unlike UF MMPs, gelatinolytic activity of these MMPs did not show distinct changes throughout the cycle. Either an inhibitor of MMP, 1,10-phenanthroline, or a metal chelator, EDTA, abolished the appearance of the above MMP activities in gelatinated gel whereas a serine proteinase inhibitor, phcnylmethylsulfonyl fluoride, failed to inhibit the appearance of MMP activities, proving that gelatinolytic activity of the above reproductive tissues were due to the enzymatic activity of MMP. When gclatinolytic activity of mouse serum was examined, it revealed 5 MMPs (131kDa, 106kDa, 89kDa, 64kDa and 62kDa bands) and one gelatinase (84kDa) band. From these results, it is concluded that the protein expression of MMPs of mouse reproductive organs, particularly uterus, is temporally regulated during estrous cycle and uterine 106kDa, 64kDa and 62kDa MMPs are suggested to play an important role in cyclic tissue remodeling of mouse uterus.

• 한국동물학회지
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• 제33권1호
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• pp.108-118
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• 1990

혈청을 비롯해서 extracellular matrix에 존재하는 당단백질인 fibronectin은 근세포의 융합과 밀접한 관계가 있는 것으로 알려져 있다. 본 연구실에서는 최근에 근세포가 분화하는 동안에 fibronectin의 수준이 감소되며, 이러한 감소는 fibronectin의 28 kDa fragment에 대한 수용체으 유용성이 감소하는 결과로 밝혀낸 바 있다. 본 연구에서는 근세포으 융합을 억제하는 물질로 알려진 EGTA를 이용하여 근세포의 융합과 28 kDa fragmen receptor의 관계를 검토하여 보았다.EGTA를 처리한 경우 EGTA를 처리하지 않은 근세포에 비해서 fibronectin의 수준과 28 kDa fragmen binding이 훨씬 적게 감소하였으며, 융합이 봉쇄된 근세포에서 EGTA를 제거하여 융합을 재개시키면 fibronectin의 수준과 28 kDa fragmen의 binding이 정상 근세포 수준으로 환원되었다. 이상의 실험 결과로 볼 때 28 kDa fragmen에 대한 수용체의 감소 또는 변화가 근세포의 분화과정에서 일어나는 fibronectin 수준의 감소와 연관성이 있음을 알 수 있다. 한편, 배양액 내에 trypsin을 처리한 경우에는 처리하지 않은 경우에 비해서 28 kDa fragmen의 binding이 현저하게 감소되었고, gangliosides를 처리한 상태에서는 gangliosides의 농도에 정비례해서 28 kDa fragmen의 binding이 감소되었다. 이 밖에 gel overlay technique을 이용하여 28 kDa fragmen가 SDS-PAGE gel에서 분자량이 약 43kDa인 단백질 및 gangliosides와 binding하는 사실을 알 수 있었다. 이러한 실험 결과를 종합하여 볼 때, 근세포의 융합은 28 kDa fragmen에 대한 receptor의 감소와 관계가 있으며, 그 수용체는 gangliosides와 비슷한 당을 가지고 있는 당단백질일 것으로 추정된다.

• 한국미생물·생명공학회지
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• 제23권6호
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• pp.730-736
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• 1995

The solubilization and proteolytic process of $\delta $-endotoxin was analvsed to compare the biochemical property of the toxin isolated from B. thuringiensis subsp. sotto. The purified crystals were dissolved in 50 mM carbonate buffer containing 10 mM dithiothreitol at pH 10 for various times. The electrophoretic pattern showed that a rapid disappearance of 138 kDa protein band. This disappearance of protein with high molecular weight was accompanied by the appearance of new protein fragment with 104 kDa, 60 kDa, and 25 kDa. For proteolvtic processing, the soluble crystals were digested with trypsin for various times. The soluble crystal protein of 104 kDa was completely disappeared. However, the protein fragment of 60 kDa and 25 kDa still remained after complete proteolysis. The comparative immunoblot analysis showed that the antiserum against intact crystals showed strong immunoreactivity to the homologous inclusion protein of 138 kDa, 104 kDa, and 25 kDa, and to the intact spores of 221 kDa and 138 kDa, but not to the vegetative cell homogenate. The sera against crystals and spores had no immunoreactivity to the vegetative cell homogenate.

• Biomolecules & Therapeutics
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• 제4권4호
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• pp.364-372
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• 1996

The primary skin and eye irritation test of 0.3% DA-5018 cream, a capsaicin analogue, were carried out in rabbits. As control materials, Zostrix-HP cream (0.075% capsaicin cream) and the base of 0.3% DA-5018 cream were used in the same manners. In the primary skin irritation test, the Primary Irritation Index (P.I.I.) was 1.6, 1.9 and 0.5 in groups treated with 0.3% DA-5018 cream, Zostrix-HP cream and the base of 0.3% DA-5018 cream, respectively. The irritation ratings of 0.3% DA-5018 cream and Zostrix-HP cream were mildly irritating. The base of 0.3% DA-5018 cream was evaluated as a non-irritating material. In the eye irritation test, 0.3% DA-5018 cream and Zostrix-HP cream could be considered as mildly irritating materials. But, the base of 0.3% DA-5018 cream was a non-irritating material. These results suggest that 0.3% DA-5018 cream has mildly irritating activity and its irritancy is similar to that of Zostrix-HP cream.

• Biomolecules & Therapeutics
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• 제1권2호
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• pp.244-250
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• 1993

Hematotoxicity and vascular irritation of DA-125, a new anthracycline antitumor antibiotic, were investigated in mice and rabbits. In hematotoxicity study, healthy male ICR mice were treated with DA-125 by a single intravenous injection at doses of 18 and 24 mg/kg. After 4, 8, 12 and 16 days WBC count, RBC count, hemoglobin concentration, hematocrit value, and platelet counts were measured respectively. As a positive control, 12 mg/kg of doxorubicin (DXR) was used in the same manner. Remakable reductions of WBC counts in groups treated with DA-125 or DXR were observed 4 days after administration and returned to normal range 8 days after injection in groups of DA-125 18 mg/kg and DXR 12 mg/kg. The recovery of leukopenia induced in a group of DA-125 24 mg/kg took about 16 days after administration. The RBC counts, hemoglobin concentrations and hematocrit values also decreased in all drug treated groups on day 8 and recovered thereafter. The platelet counts of groups treated with DA-125 or DXR decreased on day 4 and recovered from day 8 of experiment. Local vascular irritation of DA-125 was also assessed in rabbits. The obtained results can be summarized as follows. 1. Thrombophlebitis was not induced even after daily intravenous administration of 0.4% solution of DA-125 or 0.2% solution of DXR for 7 days. 2. Macro- and microscopic observations revealed that the irritative activity of 0.4% solution of DA-125 in blood vessels was not so much different from that of saline when they were injected once a day into vein retroauricularis of rabbits for 7 days. 3. Mild inflammatory reaction was noted around vessels in rabbits treated with 0.2% solution of DXR after consecutive intravenous infusion for more than 5 days. 4. The potencies of vascular irritation of the test solutions were summarized in the following order; saline = 0.4% DA-125<0.2% DXR. These results indicated that DA-125 showed similar pattern of hematotoxicity with DXR but was less hematotoxic than DXR, and that 0.4% solution of DA-125 did not elicit unusual toxic properties when injected through intravenous route for clinical practice.

• 대한약리학회지
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• 제28권1호
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• pp.11-18
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• 1992

Physostigmine을 급성(0.75 mg/kg), 7일간 매일 1회(0.75 mg/kg) 그리고 7일간 연속(0.15 mg/kg/h) 투여한 쥐의 선조체에 있어서 dopamine(DA) 및 대사체와 tyrosine hydroxylase(TH)활성도의 변화를 검색하였다. 급성 투여 1시간 후 선조체의 DA 농도는 변화가 없었으나 dihydroxy-phenylacetic acid(DOPAC)과 homovanillic acid(HVA)농도는 증가하였다. 또한 DA의 turnover의 증가를 제시하는 DOPAC/DA와 HVA/DA의 비율도 증가하였다. 그러나 TH활성도는 24시간 후 감소를 나타내었다. Physostigmine을 매일 및 연속 투여군에서는 DA과 대사체의 농도에는 변화가 없었으나, DA의 turnover의 감소를 제시하는 DA과 대사체의 비율은 감소하였다. 그러나 TH활성도는 매일 투여군에서만 감소를 나타내었다. 이러한 결과는 physostigmine을 급성 및 아급성으로 투여시 dopamine대사의 변화가 있음을 제시한다. 또한 콜린 신경계의 아급성 자극은 dopamine 대사 및 활성도를 감소 시킬 가능성을 시사해 주었다.

• Biomolecules & Therapeutics
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• 제2권4호
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• pp.310-315
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• 1994

The pharmacokinetics and tissue distribution of DA-3030 (recombinant human granulocyte colony-stimulating factor, rhG-CSF, recently manufactured by Dong-A research laboratory of Dong-A Pharmaceutical Company) were compared with reported data in the literature. After intravenous(i.v.) administration of DA-3030, at dose of 5, 10 and 100 $\mu\textrm{g}$/kg to rats, some pharmacokinetic parameters, such as terminal half-lives(1.05, 1.19 and 1.83 hr, respectively) and clearance (84.0, 54.8 and 45.5 mι/hr/kg, repectively), were dose-dependent. This could be due to the saturable metabolism of DA-3030 in rats. Similar results were also reported. After subcutaneous(s.c.) and intramuscular(i.m.) administrations of DA-3030, 10 $\mu\textrm{g}$/kg to rats, the extent of bioavailability(absolute bioavailability) were incomplete; the values were 23.3 and 18.2% after s.c. and i.m. injections, respectively, due to the degradation of DA-3030 by protease. After 7-consecutive day i.v. administrations of DA-3030, 10 $\mu\textrm{g}$/kg/day, to rats, the plasma concentrations and pharmacokinetic parameters of DA-3030 were not significantly different from those in single administration. In mice and dogs at DA-3030 dose of 10 $\mu\textrm{g}$/kg, the plasma concentrations of DA-3030 were also declined rapidly with terminal half-lives of 1.31 and 1.15 hr, respectively. DA-3030 was highly concentrated in the kidney after i.v. administration of DA-3030, 10 $\mu\textrm{g}$/kg, to rats, and the results were similar to those obtained using radiolabelled rhG-CSF in the literature. Above data indicate that DA-3030 has similar properties to rhG-CSF manufactured by other companies in view of pharmacokinetics.

• 대한수의학회지
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• 제38권3호
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• pp.518-524
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• 1998

신장 근위세뇨관세포들은 사구체에서 여과된 물질의 재흡수, 분비 및 대사에 관여하는 여러 호르몬들의 수용체들을 가지고 있다. 이들중에서 dopamine(DA)과 angiotensin II(ANG II)가 $Na^{+}/H^{+}$ 상호운반계 조절에 중요한 역할을 하고 있다. 본 연구는 초대배양한 토끼 신장 근위세뇨관세포의 $Na^+$ uptake에 있어서 DA과 ANG II의 상호관계를 알아보고자 실시하였다. DA은 농도의존적으로 $Na^+$ uptake를 유의성 있게 억제하였다($10^{-6}M$ ; $83.2{\pm}7.2%$, $10^{-3}M$ ; $67.2{\pm}3.8%$ vs. control)(p<0.05). $DA_1$ 작동제(SKF 38393, $10^{-6}M$)는 대조군의 $81.4{\pm}6.7%$ 까지 $Na^+$ uptake를 유의성 있게 억제하였으나(p < 0.05) $DA_2$ 작동제는 영향을 미치지 않았다. $DA_1$ 길항제(SCH 23390, $10^{-6}M$)에 의해 DA의 $Na^+$ uptake 억제효과는 차단되었으나 $DA_2$ 길항제(spiperone, $10^{-6}M$)에 의해서는 영향을 받지 않았다. DA과 대조적으로 $10^{-11}M$ ANG II는 $AT_1$ 수용체를 통하여 대조군의 $120.7{\pm}4.9%$까지 $Na^+$ uptake를 유의성 있게 촉진하였다. (p < 0.05). DA 및 $10^{-11}M$ ANG II를 병합처리하였을 때 DA은 농도의존적으로 ANG II에 유도된 $Na^+$ uptake 촉진효과를 유의성 있게 차단하였다(p<0.05). 한편 ANG II에 의해 유도된 $Na^+$ uptake촉진작용은 $DA_1$ 또는 $DA_2$ 작동제에 의해 차단되었으나 DA에 의한 차단 효과는 $DA_1$ 및 $DA_2$ 길항제를 병합처리하였을 때만 반전되었다. 결론적으로 DA은 $DA_1$ 수용체를 통하여 $Na^+$ uptake를 억제하였으나 ANG II에 의한 $Na^+$ uptake 촉진작용의 억제에는 $DA_1$ 및 $DA_2$ 수용체 모두가 관여하였다.