• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제40권1호
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• pp.3-10
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• 2014

Objectives: Although nerve growth factor (NGF) could promote the functional regeneration of an injured peripheral nerve, it is very difficult for NGF to sustain the therapeutic dose in the defect due to its short half-life. In this study, we loaded the NGF-bound heparin-conjugated fibrin (HCF) gel in the NGF-delivering implants and analyzed the time-dependent release of NGF and its bioactivity to evaluate the clinical effectiveness. Materials and Methods: NGF solution was made of 1.0 mg of NGF and 1.0 mL of phosphate buffered saline (PBS). Experimental group A consisted of three implants, in which $0.25{\mu}L$ of NGF solution, $0.75{\mu}L$ of HCF, $1.0{\mu}L$ of fibrinogen and $2.0{\mu}L$ of thrombin was injected via apex hole with micropipette and gelated, were put into the centrifuge tube. Three implants of experimental group B were prepared with the mixture of $0.5{\mu}L$ of NGF solution, $0.5{\mu}L$ HCF, $1.0{\mu}L$ of fibrinogen and $2.0{\mu}L$ of thrombin. These six centrifuge tubes were filled with 1.0 mL of PBS and stirred in the water-filled beaker at 50 rpm. At 1, 3, 5, 7, 10, and 14 days, 1.0 mL of solution in each tubes was collected and preserved at $-20^{\circ}C$ with adding same amount of fresh PBS. Enzyme-linked immunosorbent assay (ELISA) was done to determine in vitro release profile of NGF and its bioactivity was evaluated with neural differentiation of pheochromocytoma (PC12) cells. Results: The average concentration of released NGF in the group A and B increased for the first 5 days and then gradually decreased. Almost all of NGF was released during 10 days. Released NGF from two groups could promote neural differentiation and neurite outgrowth of PC12 cells and these bioactivity was maintained over 14 days. Conclusion: Controlled release system using NGF-HCF gel via NGF-delivering implant could be an another vehicle of delivering NGF to promote the nerve regeneration of dental implant related nerve damage.

• 농업생명과학연구
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• 제44권2호
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• pp.57-66
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• 2010

자색고구마 추출물의 항산화 효과와 산화적 스트레스로 유도된 PC12 신경세포에 대한 보호효과에 대하여 연구하였다. 자색고구마 추출물의 총 페놀함량은 44.25 mg/g, monomeric anthocyanin 함량은 2,394 mg/L로 나타났다. 자색고구마 추출물의 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis-(3-ethylben-zthiazoline-6-sulfonic acid) (ABTS) radical 소거활성, ferric reducing/antioxidant power (FRAP) 및 환원력은 농도 의존적으로 항산화 활성이 증가하였다. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazoliumbromide} reduction assay를 이용하여 자색고구마 추출물의 신경세포 보호효과를 측정한 결과, 세포 생존율이 두드러지게 증가하는 것으로 나타났다. 산화적 스트레스는 신경세포막 손상 정도를 증가시키기 때문에 lactate dehydrogenase (LDH) release assay와 neutral red uptake assay를 이용하여 세포막 손상 보호효과를 조사한 결과 자색고구마 추출물 처리구는 대조구에 비하여 산화적 스트레스로 유도된 세포막 손상 보호효과가 농도 의존적으로 나타났다. 따라서 자색고구마 추출물은 천연 항산화 소재 및 알츠하이머성 치매와 같은 신경퇴행성 질환의 예방 소재로서의 활용 가능성이 기대된다.

• 동의생리병리학회지
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• 제27권1호
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• pp.92-98
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• 2013

Bee Venom(below BV) has been used in alternative medicine to treat the diseases, such as pain diseases. BV contains a variety of peptides, including melittin, apamin, adolapin, MCD peptide, enzymes(i.e. PLA2), amines(i.e. histamine and epinephrine), and nonpeptide components. The two main components of BV are melittin and PLA2. The cell cytotoxic effects through the activation of PLA2 by melittin have been suggested to be the critical mechanism for the depress of cancer cell. Melittin and PLA2 have been reported to induce apoptosis and to possess anti-cancer effects and neurite outgrowth in PC12 cells. Analysis of proliferation was confirmed by MTT assay. BV decreased cell number through dose- and duration-dependent manner and these effects are apparent at a concentration of 3 ${\mu}g/ml$. To observe which signaling molecules will be activated by BV, phosphorylation of ERK, p38 MAPK, JNK and ERM were examined by Western blot analysis. To study the long term effect of BV in human gastric adenocarcinoma cell lines, the image of cells treated with BV for 4 days were obtained. BV was shown to exhibit anti-cancer activity in human gastric adenocarcinoma cell lines at a broad range of concentrations of 3 ${\mu}g/ml$. ERK, p38 MAPK and JNK were found to increase in BV treated cells. However, ERM which known to be involved in the cell death, was gradually decreased to 30minutes after addition 3 ${\mu}g/ml$ of BV. These results provide a possible BV-induced inhibitory signal for cancer proliferation that is initiated by the decrease in ERM activity. Moreover, it is likely that the activation of ERK, p38 MAPK and JNK are required for the BV-induced inhibition of cancer proliferation.

• Fisheries and Aquatic Sciences
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• 제3권1호
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• pp.26-32
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• 2000

Monospecific red tide by a toxic dinoflagellate belonging to the genus Alexandrium occurred at Chindong Bay in the southern coast of Korea and continued from April 6th to 15th in 1997. The ratio of its cell number to total phytoplankton cell number was much higher than $95\%$. This organism was identified as Alexandrium tamarense, although slight morphological differences were found comparing to the original and successive descriptions of the species. We found neither anterior nor posterior attachment pores in these cells of the bloom population. The occurrence of red tide caused by A. tamarense was first reported in Korea. Its plate formula is Po, Pc, 4', 6"c, 8s, 5"' and 2"". Thecal plates are thin with pore-like ornamentation. In those plates, the anterior part of the first apical plate (1') is narrower and its posterior end has sometimes a block-like accessory, but this variation was considered within the range of the morphological variability of this taxon. The cell density during the red tide exhibited a wide range of variation by the depth of water column, ranging from $2\times10^6$ cells$l^{-1}$ to $5\times10^6$ cells·$l^{-1}$. Water temperature varied from 11.8 to $12.3^{\circ}C$. Toxicity of A. tamarense during red tide was measured as $8.8\times10^5$. $MU\;\cdot\;cell^{-1}$ by mouse bioassay.

• Toxicological Research
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• 제20권3호
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• pp.241-250
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• 2004

We previously found that bisphenol A (BPA) caused neurotoxic behavioral alteration. Since disturbance of calcium homeostasis is an implicated contributor in the neurotoxic mechanism of environmental toxicants, we investigated whether BPA alters calcium homeostasis. Unlike other neurotoxic agents which cause increase of intracellular calcium level, BPA decreased $[Ca^{2+}]_i$ dose-dependently in PC12 cells and cortical neuronal cells regardless of the calcium existence in buffer. BPA at greater concentrations than 100 $\mu\textrm{M}$ reduced cell viability significantly in both types of cells. BPA also suppressed L-glutamate (L-type channel activator, 30 mM) and trifluoperazine (calmodulin antagonist, 30 $\mu\textrm{M}$)-induced increase of $[Ca^{2+}]_i$. BPA further lowered caffeine (RYR activator, 100 $\mu\textrm{M}$)-decreased $[Ca^{2+}]_i$, but did not alter dantrolene (RYR inhibitor, 100 $\mu\textrm{M}$), heparin (IP3 inhibitor, 200 units/ml) and xestospongin C (IP3 inhibitor, 5 $\mu\textrm{M}$)-decreased $[Ca^{2+}]_i$. Cell viability was not directly related to intracellular calcium change by bisphenol A that alternation of intracellular calcium may not be a direct causal factor of BPA-induced neuronal cell death.

• 한국환경보건학회지
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• 제23권2호
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• pp.12-23
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• 1997

Potassium dichromate(PD, 500 ppm, 500 ppm), potassium chromate(PC, 500 ppm, 500 ppm), cobalt chloride(CC, 100 ppm, 10 ppm), methylmercuric chloride(MC, 100 ppm, 10 ppm)을 처리한 배지에 E. coli와 B.subtilis를 배양하는 동안에 당지질과 galactose의 생합성 및 지방산 조성을 대조구와 비교 분석하였다. 금속화합물 처리구에서의 생장과 MGDG, DGDG의 함량 및 total lipid는 대조구에 비해 감소하였다. 또한 당지질 형성에 이용된 galatose의 함량은 억제되었다. E. coli와 B. subtilis에서 MGDG와 DGDG 합성에 이용된 주요 지방산은 다양한 변화를 나타내었다.

• The Korean Journal of Physiology and Pharmacology
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• 제14권3호
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• pp.163-167
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• 2010

This study investigated the effects of the methanol extracts of Morinda citrifolia containing numerous anthraquinone and iridoid on phospholipase $A_2$ ($PLA_2$) isozyme. $PLA_2$ activity was measured using various $PLA_2$ substrates, including 10-pyrene phosphatidylcholine, 1-palmitoyl-2-[$^{14}C$]arachidonyl phosphatidylcholine ([$^{14}C$]AA-PC), and [$^3H$]arachidonic acid (AA). The methanol extracts suppressed melittin-induced [$_3H$]AA release in a concentration-dependent manner in RAW 264.7 cells, and inhibited $cPLA_2/sPLA_2$-induced hydrolysis of [$^{14}C$]AA-PC in a concentration- and time-dependent manner. A Dixon plot showed that the inhibition by methanol extracts on $cPLA_2$ and $sPLA_2$ appeared to be competitive with inhibition constants ($K_i$) of $3.7{\mu}g/ml$ and $12.6{\mu}g/ml$, respectively. These data suggest that methanol extracts of Morinda citrifolia inhibits both $Ca^{2+}$-dependent $PLA_2$ such as, $cPLA_2$ and $sPLA_2$. Therefore, Morinda citrifolia may possess anti-inflammatory activity secondary to $Ca^{2+}$-dependent $PLA_2$ inhibition.

• Animal Bioscience
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• 제36권8호
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• pp.1252-1262
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• 2023

Objective: Microencapsulation technologies have been developed and successfully applied to protect the probiotic bacterial cells damaged by environmental exposure. This study aimed to investigate the effects of microencapsulation of Lactobacillus plantarum MB001 on the growth performance, ileal nutrient digestibility, jejunal histomorphology and cecal microbiome of broiler chickens in a tropical climate. Methods: A total of 288 one-day-old female broilers (Ross 308) were randomly allocated into 4 groups (6 replicates of 12 birds). Treatments included, i) a basal diet (NC), ii) NC + avilamycin (10 mg/kg) (PC), iii) NC + non-encapsulated L. plantarum MB001 (1×10⁸ colony-forming unit [CFU]/kg of diet) (N-LP), iv) NC + microencapsulated L. plantarum MB001 (1×10⁸ CFU/kg of diet) (ME-LP). Results: Dietary supplementation of ME-LP improved average daily gain, and feed conversion ratio of broilers throughout the 42-d trial period (p<0.05), whereas ME-LP did not affect average daily feed intake compared with NC group. Both N-LP and ME-LP improved apparent ileal digestibility of crude protein and ether extract compared with NC group (p<0.05). The broilers fed ME-LP supplemented diet exhibited a beneficial effect on jejunal histomorphology of villus height (VH), crypt depth (CD) and villus height to crypt depth ratio (VH:CD) of broilers compared to NC group (p<0.05). At the phylum level, Firmicutes was enriched (p<0.05) and Proteobacteria was decreased (p<0.05) only in the ME-LP group. At the genus level, the ME-LP diets increased (p<0.05) the number of both Lactobacillus and Enterococcus compared to NC, PC, and N-LP groups (p<0.05). Conclusion: Microencapsulation assists the efficient functioning of probiotics. ME-LP could be potentially used as a feed additive for improvement of cecal microbiota, gut integrity and nutrient utilization, leading to better performance of broilers.

• 한국산학기술학회논문지
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• 제18권12호
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• pp.575-580
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• 2017

본 연구는 hnRNP A1 단백질에 포함되어 있는 RGG 상자가 이 단백질의 세포내 위치에 미치는 영향 및 이 단백질의 안정화에 미치는 영향을 분석하는 것을 목적으로 하며 2014년 10월부터 약 3년 동안 수행되었다. 이를 위해 먼저, RGG상자 내의 6개의 아르기닌을 라이신으로 돌연변이를 만든 다음 pcDNA1-HA-hnRNP A1(6R/K)를 재조합하였다. 다음, 이 플라스미드 DNA를 HeLa 세포에 형질주입하여 HA-hnRNP A1(6R/K) 단백질의 세포내 위치에 미치는 영향을 면역형광현미경법으로 분석하였다. 그 결과 hnRNP A1(6R/K) 단백질은 (wt 단백질처럼 핵에 위치하지 못하고) 핵과 세포질 모두에 퍼져 있는 것을 확인하였다. hnRNP A1(6R/K)의 안정성을 조사하기 위해서는 pcDNA1-HA-hnRNP A1(6R/K)를 HeLa 세포에 형질 주입시킨 다음 발현된 단백질을 웨스턴 블랏 실험으로 분석하였는데, 그 결과 HA-hnRNP A1(6R/K)는 (잘려서) 크기가 작아진 것을 확인할 수 있었다. 이 결과들로부터 HA-hnRNP A1(6R/K)가 핵과 세포질 모두에 퍼져 있는 것은 6R/K 돌연변이가 hnRNP A1의 핵 위치 능력에 영향을 미쳤기 때문이 아니라 6R/K 돌연변이가 일어난 부위 또는 그 부근이 절단되어 hnRNP A1의 핵 위치 신호(nuclear localization singal)인 M9 도메인이 들어있는 C-말단 부위를 잃었기 때문이라는 점을 확인 할 수 있었다. 본 연구의 결과들은 hnRNP A1에 있는 RGG 상자의 아르기닌이 hnRNP A1의 안정성에 중요한 역할을 한다는 것을 제시한다. 세균에서 발현시켜서 정제된 hnRNP A1(6R/K)의 RNA-결합 능력에 대한 영향 분석은 추후 과제로 남겨둔다.

• Tuberculosis and Respiratory Diseases
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• 제44권4호
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• pp.822-835
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• 1997

연구배경 : 기관지천식의 병태생리중 IgE의 합성 및 조절에 IL-4가 중요한 역할을 하며, IFN-$\gamma$는 이러한 IL-4의 작용을 길항하는 것으로 알려져 있다. 이를 근거로 최근에 IFN-$\gamma$를 아토피 피부염등 IgE가 높은 알레르기성 질환의 치료에 이용하고자 하는 임상적 시도가 있는데, 아직 기관지천식에 대한 임상적 시도는 별로 보고되어 있지 않다. 그러나 IFN-$\gamma$는 in vitro에서 말초혈액 다형핵구나 단핵세포에 의한 독성산화물 생성을 증가시킨다는 보고도 있어, 실제 임상에서 IFN-$\gamma$ 치료의 임상적 효과는 확실하지 않다. 이에 저자들 IL-4 매개성 IgE 생성에 대한 IFN-$\gamma$의 길항작용에 기초하여, IgE가 정상범위보다 높은 기관지천식 환지들에서 in vitro 에서와 같이 in vivo 에서도 IFN-$\gamma$가 IgE 생성을 억제하는지 여부와, 그 결과로 임상적 치료효과를 나타내는지에 대해 관찰하는 한편, 다형핵구의 독성산화물 생성능에 미치는 영향은 in vitro의 경우와 어떻게 다른지에 대해서 관찰하였다. 대상 및 방법 : IgE가 200IU/ml 이상이고 정규적인 부신피질호르몬 치료에 반응을 보이지 않는 기관지천식 환자 50명과 정상인 17명을 대상으로, 혈중 CD23+ B-상층액 발현도, sCD23 농도, T-상층액의 IL-4 activity, 다형핵구에 의한 과산화 음이온 생성능등을 측정한 후, 환자군과 정상군간의 차이를 비교하고, 환자군에 대해서는 상기 검사외에 혈중 IgE 농도 및 histamine $PC_{20}$등을 함께 측정하여 IFN-$\gamma$ 치료전후의 변화를 관찰하였다. IFN-$\gamma$ 치료는 체중당 30,000IU를 매일 4주간 피하주사하였다. 결 과 : 다형핵구의 ${O_2}^-$ 생성능 환자군의 다형핵구는 정상군의 다형핵구에 비해 ${O_2}^-$생성능이 높았다(P<0.05). IFN-$\gamma$ 치료후 추적이 가능했던 환자들에서는 IFN-$\gamma$ 치료후에 다형핵구의 ${O_2}^-$ 생성능이 현저하게 감소하였다(P<0.05). 다형핵구 배양시간에 따라 자연적인 ${O_2}^-$ 생성능 및 PMA 혹은 fMLP 자극에 의한 ${O_2}^-$ 생성능을 관찰하였을 때에도, 배양시간 및 자극제의 종류와 무관하게 IFN-$\gamma$ 치료후가 치료전에 비해 ${O_2}^-$ 생성능이 감소하는 경향을 보였다. IFN-$\gamma$ 치료전의 환자군의 말초혈액내 CD23+ B-상층액의 발현도는 정상군에 비해 현저히 높았으나(P<0.05), 치료후 추적검사가 가능하였던 15명의 환자들에서 치료전후에 의미있는 변화를 나타내지는 않았다. IFN-$\gamma$ 치료전의 환자군의 혈청내 sCD23 농도는 정상군에 비해 다소 높은 경향을 보였으며, 이중 치료 후 추적검사가 가능하였던 17명의 환자들중 11명(64.7%)에서 치료후에 혈청 sCD23 농도가 감소되었다. T-상층액의 IL-4 activity는 정상군에 비해 환자군에서 다소 높은 경향을 보였는데, IFN-$\gamma$ 치료후에는 큰 변화를 보이지 않았다. IFN-$\gamma$ 치료후 혈청 IgE 농도의 추적검사가 시행되었던 환자 15명 중 9명(60%)에서 치료후 혈청 IgE 농도가 유의하게 감소되었다(P<0.05). IFN-$\gamma$ 치료에 따른 기관지과민반웅의 변화는 모두 12명의 환자에서 관찰되었는데, 이중 10명(83.3%)에서 IFN-$\gamma$ 치료후 histamine $PC_{20}$가 유의하게 호전되는 소견을 보여주었다(P<0.05). 결 론 : 이상의 연구결과 IFN-$\gamma$는 IgE가 높은 기관지천식 환자의 치료에 유용할 것으로 추측되며, in vivo에서의 IgE 합성 및 조절기전과 이에 대한 IFN-$\gamma$의 역할을 보다 명확하게 규명하기 위한 노력이 필요할 것으로 생각된다.