• Journal of Preventive Medicine and Public Health
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• 제26권4호
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• pp.565-573
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• 1993

Noise is not only affecting the ear and the auditory cortex locally, but its influence is widely spread throughout the brain structures, e. g., the reticular formation, the brain stem nuclei or the subcortical forebrain area. Hence, any of the organism's activities can be hindered or stimulated by noise. High noise is a stressor and the catecholamine level can be used both as a stress marker and as an indicator of modified sympathetic nervous system activity. Several recent studies have found that the urinary excretion of catecholamines is increased due to high noise intensity, especially unexpectedly high and long lasting noise. The present study was conducted in order to examine the effects of noise stress on urinary excretion of ctecholamines in rats and humans. Rats were exposed to 90 dB noise for 10, 30, and 60 minutes, 3 and 12 hours. 24 hour . urinary samples were collected and the catecholamones were extracted by alumina and analyzed by HPLC-ECD. Catecholamine levels increased with time of exposure up to 60 minutes : norepinephrine concentration at 60 min of noise=1.038 ng/ml, epinephrine=0.636 ng/ml. Urine catecholamines of blue collar workers exposed to 90 dB of noise at the work place were collected between 2 and 4 p.m. and compared to that of white collar workers exposed to 70 dB. Mean norepinephrine level of the blue collar workers was 0.89 ng/ml (${\pm}0.25$), epinephrine 0.24ng/m1 (${\pm}0.09$), and that of the white collar workers 0.48 ng/ml (${\pm}0.12$), epinephrine 0.19 ng/ml(${\pm}0.05$). It was concluded that noise acts as a stressor and increases the catecholamine levels in both rats and humans.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.135-141
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• 2010

Embryoid bodies (EBs) generated from human embryonic stem cells (hESCs) include spontaneously induced endodermal lineage cells (ELCs). Activin-A plays important roles in the endoderm differentiation of hESCs. Despite studies on the generation of ELCs from hESCs with treatment of Actvin-A, it was unclear for localization and pattern of ELCs by Activin-A during differentiation of hESCs. Accordingly in this study, we knew that Actvin-A increased the cystic EBs formation, including the highly enriched AFP (endoderm lineage specific marker)-expressing cells in the surface of cystic EBs. To induce the EBs formation from undifferentiated hESCs, cells were transferred onto petri-dish and cultured in suspension condition without bFGF removed hESC media (EB media) for 3 days. Next to investigate the effect of Activin-A, EBs were subsequently cultured in EB media supplement with 100 ng/ml Activin-A for 3 days. After 5~7 days of Activin-A treatment, cystic EBs began to appear which increased in numbers reaching ~60% of initially formed EBs over 5 days. Endoderm lineage marker, AFP were highly expressed and specifically localized at the surface region of cystic EBs comparison with normal EBs. We next attached the cystic EBs onto gelatin-coated plates and cultured for 5 days. In the results of real-time PCR and immunocytochemistry analysis, AFP-expressing cells migrated and localized at the outgrowth region of attached cystic EBs. To obtain the AFP-expressing cells of the outgrowth region, we manually isolated by using micro-dissection and cultured them. These cells strongly express AFP over 70% of isolated cells post re-plating. Here, we first showed an expression pattern of specifically localized ELCs by Activin-A during differentiation of hESCs. From this observation, we could highly purified ELCs from undifferentiated hESCs. Taken together, our system will provide a novel and efficient option to generate ELCs from hESCs.

• Applied Biological Chemistry
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• 제48권2호
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• pp.166-172
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• 2005

제주자생 탐라오갈피(Acanthopanax koreanum Nakai)를 리큐르 소재로 활용하기 위하여 주정농도를 각각 $30{\sim}95%$로 조절한 다음 0.5 cm 이하로 세절한 뿌리 및 줄기를 700 g/10 l의 비율로 첨가하여 70일간 침출시키면서 이화학적 특성 및 유용성분의 경시적인 변화를 검토하였다. 침출액의 pH 변화는 대체로 $5.5{\sim}6.5$ 범위였으며, 색도 b 값은 주정농도가 낮을수록, 침출기간이 길어질수록 높은 값을 나타내었고, a값은 주정농도가 높을수록 - 값이 증가하였으며 주정농도에 의한 영향이 컸다. 가용성고형물의 변화는 침출기간에 따라 점진적으로 증가하여 주정농도 $30{\sim}70%$ 범위에서 줄기는 $0.6{\sim}0.7%(w/v)$, 뿌리는 $1.0{\sim}1.5%(w/v)$이었다. Eleutheroside B와 E는 침출기간 20일까지 급속히 증가하는 경향이었으며, 줄기가 뿌리보다 함량이 많았다. 그리고 주정농도 30%에서 70%로 높아짐에 따라 eleutheroside B와 E 함량은 크게 증가하였다. Acanthoic acid는 주정농도와 이용부위에 따른 함량변화가 뚜렷하게 나타났으며, 침출 후 $5{\sim}10$일에 급속히 용출되었으며, 줄기에서는 $3{\sim}25\;{\mu}g/ml$, 뿌리에서는 $46{\sim}1,700\;{\mu}g/ml$이 침출되었다. 따라서 탐라오갈피를 이용하여 약용주인 리큐르로 상품화하기 위해서는 뿌리 비율을 높게 하여 주정농도 $60{\sim}80%$ 범위에서 $30{\sim}50$일간 침출시킨 다음 13주간 숙성하여 블렌딩하는 것이 효과적이라고 판단되었다.

• 생명과학회지
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• 제34권3호
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• pp.160-169
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• 2024

본 연구에서는 분비나무(Abies nephrolepis MAX.)를 분비나무 줄기(Abies nephrolepis MAX. stem) 추출물과 분비나무 잎(Abies nephrolepis MAX. leaf) 추출물로 나누어 항염 관련 기능성을 연구하여 화장품 소재로서의 이용가능성을 알아보고자 하였다. MTT assay를 통해 세포 생존율을 확인한 결과, 대식세포(RAW 264.7)에서 분비나무 줄기와 잎 추출물의 500 ㎍/ml의 농도에서 각각 97.8%, 95.6%의 생존율을 보였다. 이에 따라 세포 관련 실험을 세포의 생존율이 100%에 가까운 25, 50, 100 ㎍/ml의 농도로 진행하였다. 염증을 유발하는 활성산소인 NO 생성에 대해 분비나무 추출물을 측정한 결과, 분비나무 추출물을 처리한 구에서 NO 발현이 억제되는 것을 확인할 수 있었으며, 분비나무 줄기와 잎은 100 ㎍/ml의 농도에서 71.1%, 42.9%의 저해율을 나타내었다. 분비나무 추출물을 RAW 264.7 세포에 처리하여 단백질 발현을 측정한 결과, COX-2, iNOS 및 pro-inflammatoty cytokine인 IL-1β, IL-6, TNF-α 인자에서 농도 의존적으로 단백질 발현이 억제되었으며, 분비나무 줄기의 iNOS와 분비나무 잎의 COX-2, iNOS, IL-6 및 TNF-α의 인자가 대조군인 Vit. C와 비교하였을 때 우수한 단백질 발현 억제를 보였다. 또한, RAW 264.7 세포에 분비나무 추출물을 처리하여 mRNA 발현을 측정한 결과, 분비나무 줄기의 iNOS, 분비나무 잎의 COX-2와 iNOS의 mRNA 발현양은 대조군인 Vit. C와 비교하였을 때 우수한 억제율을 확인할 수 있었으며, pro-inflammatoty cytokine인 IL-1β, IL-6 및 TNF-α인자에서는 분비나무 줄기 추출물의 IL-1β는 대조군과 유의한 결과를 나타냈지만, 나머지 인자에서는 대조군인 Vit. C와 비교하였을 때 mRNA 발현 억제율이 우수한 것을 확인할 수 있었다. 이에 따라 분비나무 추출물이 항염 관련 기능성 활성에 우수함을 나타내어 화장품 소재로써의 이용가능성을 확인할 수 있었다.

• 한국식물병리학회지
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• 제14권4호
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• pp.339-344
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• 1998

Since the leaf inoculation procedures are time-consuming and require considerable growth chamber space, a rapid dioassay method for screening of pathogenicity of Didymella bryoniae, a casual agent of gummy stem blight in watermelon, was established in this paper. The method produced reliable results within 8 days ( 5 days for growing seedlings and 3 days for rapid disease response in the seedlings). After contaminants in the root of 4~5 day-old seedlings had been washed using sterilized water, 5 seedlings were dipped into a vial containing 12 ml of conidial suspension (106 cells/ml). After the vials were placed in a growth chamber (22$^{\circ}C$, RH 50%, 14hr light/10hr darkness) for 3 days, susceptibility and resistance of cultivars were determined by the degree of disease response on cotyledon. The result of obtained by the dip-inoculation method was well coincided with the results by the leaf inoculation procedures and the result that had been observed for several years in the field. Screening of collected watermelon cultivars by the dip-inoculation method revealed that all the 21 domestic cultivars collected were susceptible and only 3 foreign cultivars (PI 189225, PI 482322 and IT 188207) were resistant among 18 cultivars A cucumber cultivar (Marketer) and bitter cucumber were proven to be resistant against the D. bryoniae among 8 other different cucurbits tested. The SDS-PAGE patterns of total proteins from a susceptible (Keumcheon) and a resistant (PI 189225) watermelon cultivars were compared 0, 12, 24 and 36 hrs after inoculation. The amounts of two distinct protein bands (24 kDa and 70 kDa) were gradually increased after inoculation in both cultivars.

• Journal of Plant Biotechnology
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• 제50권
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• pp.97-107
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• 2023

Malaria remains to be one of the most severe global public health concerns. Traditionally, Aspilia Africana and Warburgia ugandensis have been used to treat malaria in several African countries for millennia. In the current study, A. africana calli (AaC), A. africana in vitro roots (AaIR), A. africana wild leaf (AaWL), and W. ugandensis stem bark (WuSB) were dried and pulverized. Fourier transform near-infrared spectroscopy was used to analyze the powdered samples, while 80% ethanolic extracts of each sample were assayed for antiplasmodial activity (against Plasmodium falciparum strains DD2 (chloroquine-resistant) and 3D7 (chloroquine-sensitive)) and cytotoxicity. WuSB showed the highest antiplasmodial activity (IC₅₀ = 1.57 ± 0.210 ㎍/ml and 8.92 ± 0.365 ㎍/ml against P. falciparum 3D7 and DD2, respectively) and selectivity indices (43.90 ± 7.914 and 7.543 ± 0.051 for P. falciparum 3D7 and DD2, respectively). The highest total polyphenolic contents (total phenolic and flavonoid contents of 367.9 ± 3.55 mg GAE/g and 203.9 ± 1.43 mg RUE/g, respectively) were recorded for WuSB and the lowest were recorded for AaC. The antiplasmodial activities of the tested plant tissues correlated positively with total polyphenolic content. The high selectivity indices of WuSB justify its traditional applications in treating malaria and present it as a good candidate for discovering new antimalarial compounds. We recommend elicitation treatment for AaIR, which showed moderate antiplasmodial activity against P. falciparum DD2, to increase its secondary metabolite production for optimal antimalarial activity.

• 한국가축번식학회지
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• 제21권2호
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• pp.157-165
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• 1997

포유류 배반포배의 epiblast는 내세포괴에 포함되어 있으며, 이 epiblast cells이 배 및 태아의 생식세포와 일반 체세포로 분화된다 (Beddington, 1986; Lawson 등, 1991). 그런데 조기에 발달된 부화배반포기 배가 지연발생된 부화배반포기 배보다도 많은 epiblast cells을 가지고 있다고 한다(Talbot 등, 1995). 그래서 본 연구에서는 체외수정 유래의 배반포배의 발육속도에 따른 내세포괴/영양배엽세포의 비율 및 배아간세포 확립 효율을 비교하여 발달일령 간에 차이가 있는지를 규명하고자 하였다. 공시한 소의 난포란을 TCM-199에 0.5$\mu\textrm{g}$/ml FSH, 5$\mu\textrm{g}$/ml LH, 10% FBS, 100 units/ml penicilin, 및 100$\mu\textrm{g}$/ml streptomycin을 첨가하여 39$^{\circ}C$, 5% CO2 조건하에서 24시간 동안 체외성숙한 후, 5$\mu\textrm{g}$/ml heparin으로 수정능이 획득된 1$\times$106 sperm/ml의 정자로 체외수정을 유도하였다. 체외수정 후 18~20시간에 과립막세포를 vortexing으로 제거하여 얻은 모든 체외수정란을 3mg/ml BSA, 20${\mu}\ell$/ml NEM amino acids, 40${\mu}\ell$/ml BME amino acids, 10mM glycine 및 1mM alanine이 함유된 CR1aa 배양액에서 BRL 단층세포와 공배양을 실시하였다. 수정후 7, 8 및 9일재 (체외수정일 : 0일)에 확장배반포기까지 발달한 수정란을 이중염색 및 배아간세포의 확립 실험에 공시하였다. 체외수정후 24시간에 분할된 총 1,145개의 수정란이 7, 8 및 9일째에 후기 배반포기까지 각각 222(15.6%), 103(7.2%) 및 52(3.6%)로 발달하여 총 377개 (26.4%)가 발달하였다. 내세포괴/영양배엽세포의 비율은 7일 및 8일째 배반포배에서 각각 47.2$\pm$11.9/95.1$\pm$24.4개 (33/67%) 및 40.3$\pm$12.4/83.3$\pm$26.9개 (33/67%)로서 9일째 배반포배의 19.3$\pm$8.1/62.3$\pm$23.1개 (24/76%) 보다 유의적(P<0.05)으로 높았다. ES-like cells을 확립하기 위하여 후기배반포기 배를 mouse embryonic fibroblast 단층 공배양기에 옮긴 후 5일에 내세포괴의 부착 여부를 판정하고, 10일에 배아간세포의 확립 여부를 판정하였다. 그 결과 7, 8 및 9일째의 배반포기배의 각각 47.7% (82/172), 30.9%(22/71) 및 15.6%(5/32)에서 배아간세포가 확립되었다. 이상의 결과에서 배반포기까지의 발육이 빠른 수정란에서 영양배엽에 대한 내세포괴세포의 비율이 높았고 배아간세포의 확립율도 높다는 사실을 입증할 수 있었으며, 이와 같은 결과에서 체외수정란 유래 배반포배의 질을 결정할 수 있을 것으로 생각된다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제35권6호
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• pp.397-402
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• 2009

The purpose of this study is to investigate the effects of alendronate and pamidronate on proliferation and the alkaline phosphatase activity of human bone marrow derived mesenchymal stem cells and to relate the results with bisphosphonate related osteonecrosis of the jaw(BRONJ). With the consent of patients with no systemic disease and undergoing iliac bone graft, cancellous bone was collected to obtain human bone marrow derived mesenchymal stem cells through cell culture. 96 well plate were prepared with a concentration of $10^4$cell/ well. Alendronate and pamidronate were added to each well with the concentration of $10^{-6}M$, $10^{-8}M$ and $10^{-10}M$, respectively. Then proliferation capacity of each well was evaluated with the cell counting kit. 24 well plates were prepared with a concentration of $10^5$cell/ml/well and with the bone supplement, alendronate and pamidronate were added with the concentration of $10^{-6}M$, $10^{-8}M$ and $10^{-10}M$, respectively on each plate. The plates were cultured for either 24 or 72 hours. Then the cells were sonicated to measure the alkaline phosphatase activity and protein assay was done to standardize the data for analysis. As the concentration of alendronate or pamidronate added to the culture increased, the proliferation capacity of the cells decreased. However, no statistical significance was found between the group with $10^{-10}M$ of bisphophonate and the control group. Pamidronate was not capable of increasing the alkaline phosphatase activity in all trials. However, alkaline phosphatase activity increased with 24 hours of $10^{-8}M$ of alendronate treatment and with 48 hours of $10^{-10}M$ of alendronate treatment. Cell toxicity increased as the bisphosphonate concentration increased. This seems to be associated with the long half life of bisphosphonate, resulting in high concentration of bisphosphonate in the jaw and thus displaying delayed healing after surgical procedures. Alendronate has shown to increase the alkaline phophatase activity of human bone marrow derived mesenchymal stem cells. However, this data is insufficient to conclude that alendronate facilitates the differentiation of human bone marrow derived mesenchymal stem cells. Further studies on DNA level and animal studies are required to support these results.

• Applied Biological Chemistry
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• 제38권2호
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• pp.123-128
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• 1995

Agrobacterium rhizogenes를 매개로 한 Populus의 형질전환계를 확립하고자 형질전환율에 영향을 주는 주요 인자들을 검토하고, 형질전환 식물체를 분화시켜 도입유전자의 발현을 확인하였다. 줄기조직보다 잎조직이 kanamycin에 대한 감수성이 커 형질전환체의 선발에 유리한 것으로 판단되었다. 접종용액의 박테리아 밀도는 $4{\times}10^5{\sim}7{\times}10^9\;cfu$의 범위내에서 형질전환율에 거의 영향을 주지 않았다. 공배양 기간은 1일 이내가 바람직하였고, 박테리아 제거배지의 항생제 농도는 cefotaxime과 ampicillin 각각 $250\;{\mu}g/ml$가 적당하였다. 배지에 acetosyringone을 첨가하여 배양한 박테리아를 사용했을 때 형질전환율이 증가했는데 acetosyringone의 적정농도는 $50\;{\mu}M$이었다. 형질전환된 gall은 kanamycin 농도가 $100\;{\mu}g/ml$ 이상인 배지를 사용하거나, 생장조절제가 들어있지 않은 배지를 사용하여 선택적으로 유기 및 증식시킬 수 있었다. A. rhizogenes를 접종시킨 조직으로부터 유기된 gall은 생장조절제를 포함하는 배지뿐만 아니라, 생장조절제가 없는 배지에서도 3주 이내에 뿌리를 형성하였다. NAA 0.05 mg/ml와 BA 0.5 mg/ml를 첨가한 배지에서 배양된 gall로 부터 약 6주일 후 식물체가 분화되었다. A. rhizogenes를 접종시켜 얻은 gall,그리고 gall로부터 재분화된 식물체의 추출물에서 agropine과 mannopine이 검출되어, 도입된 opine합성 유전자가 발현되고 있는 것이 확인되었다.

• 한국식품영양학회지
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• 제14권6호
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• pp.527-531
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• 2001

옻나무는 독성이 있는 금기물질임에도 불구하고 한국에서는 민간요법으로 옻닭 등의 가공식품 형태로 사용하여 왔다. 항산화 물질은 옻나무 껍질에 물을 가하여 추출하여 분리하였다. 물 추출물은 DEAE, CN, ODS 컬럼을 사용하여 HPLC로 정제하였다. 정제된 순수물질 안정성은 pH 3.0~6.0의 산성영역에서는 안정하였으나, pH 6.5 이상에서는 불안정하였으며, 10$0^{\circ}C$에서 5시간 열처리하여도 80%의 활성을 보였다. 항균력 실험에서는 그람양성, 음성 균주에 대하여 항균 활성이 없었다. 항산화력은 DPPH 방법으로 조사한 결과 동일한 농도에서 (20$\mu\textrm{g}$/m1) BHT, BHC 보다 좋았으나, ascorbic acid 보다 낮았다.