• 미생물학회지
• /
• 제54권2호
• /
• pp.126-135
• /
• 2018

여러 종류의 mannanase를 생산하는 Cellulosimicrobium sp. YB-43으로부터 mannanase B를 암호하는 manB 유전자와 효소의 특성이 보고된 바 있다. Mannanase C (ManC)로 명명한 효소의 유전자가 manB 유전자의 하류에 위치한 것으로 예상되어 이를 중합효소 연쇄반응으로 클로닝하여 manC 유전자의 염기서열을 결정하였다. ManC는 448 아미노산 잔기로 구성된 것으로 확인되었으며 glycosyl hydrolase family 5에 속하는 mannanase와 상동성이 높은 활성영역과 탄수화물 결합영역(CBM2)이 존재하였다. ManC의 활성영역은 Streptomyces sp. SirexAA-E (55.8%; 4FK9_A) 및 S. thermoluteus (57.6%; BAM62868)의 mannanase와 아미노산 배열의 상동성이 55% 이상으로 가장 높았다. Signal peptide 영역이 제거되고 카르복실 말단에 hexahistidine이 연결되도록 제조한 His-tagged ManC (HtManC)의 유전자를 재조합 대장균에서 발현하여 균체 파쇄액으로부터 HtManC를 정제하였다. HtManC은 $65^{\circ}C$와 pH 7.5에서 최대 활성을 보였으며 pH 7.5~10범위에서 활성에 큰 변화가 없었다. HtManC는 locust bean gum (LBG)과 konjac에 대한 분해 활성이 guar gum과 ivory nut mannan (ivory nut)에 비해 높았다. 최적 반응조건에서 LBG를 기질로 하여 반응 동력학적 계수를 측정한 결과 Vmax와 Km이 68 U/mg과 0.45 mg/ml로 나타났다. HtManC에 의한 만노올리고당(MOS)과 mannan의 분해산물을 TLC로 관찰한 결과 mannobiose 보다 중합도가 큰MOS로부터 mannobiose와 mannotriose가 주된 분해산물로 생성되었다. 또한 LBG, konjac과 ivory nut의 분해산물로 mannobiose와 소량이 mannose가 공통적으로 관찰되었다.

• 대한지역사회영양학회지
• /
• 제2권2호
• /
• pp.179-186
• /
• 1997

A new bioelectrical impedance method has been developed and evaluated. The electrodes; were made of stainless steel and electrical interfaces were created by an upright subject gripping hand electrodes and stepping onto foot electrodes. Eight tactile electrodes were in contact with surfaces of both hands and feet; thumb, palm and fingers, front sole, and rear sole. Automatic on-off switches were used to change current pathways and to measure voltage differences for target segments. Segmental body resistances and whole body resistance(RWHOLE)were measured in 60 healthy subjects. Segmental resistances of right arm(RRA), left arm(RLA), trunk(RT), right leg(RRL) and left leg(RLL)were310.0$\pm$61.6$\Omega$, 316.9$\pm$64.6$\Omega$, 25.1$\pm$3.4$\Omega$, 236.8$\pm$31.2$\Omega$ and 237.6$\pm$30.4$\Omega$, respectively. Individual segmental impedance indexes(Ht2/RRA, Ht2/RT, and Ht2 /RLA) were closely related to lean body mass(LBM)as measured by densitometry ranged from r=0.925 to 0.960. Ht2/(RRA+RT+RLA) predicted LBM slightly better(r=0.969) than the traditional index, Ht2/RWHOLE(r=0.964), supporting the accuracy of the segmental measurement. A multiple regression equation utilizing Ht2/RRA, Ht2/RT and Ht2/RRL predicted LBM with r=0.971. Ht2/RRA term of the regression contributed to more than 40$\%$ of the LBM prediction, indicating that lean mass of arm represented whole body LBM more closely than other body segments. The new bioimpedance method was characterized by upright posture, eight tactile electrodes, segmental measurements and utilization of electronic switches in comparison with the traditional method. The measurement with this new method was extremely reproducible, quick and easy to use.

• Toxicological Research
• /
• 제14권4호
• /
• pp.569-575
• /
• 1998

One of the most frequent dejects in human cancer is the uncontrolled activation of the ms-signaling pathways. Significant evidence has accumulated to directly implicate members of the matrix metalloproteinases (MMPs) in tumor invasion and metastasis formation. We have previously shown that MMP-9 expression was significantly enhanced in the ras-tranfected HT1080 human fibrosarcoma cells at the mRNA level. In the present study, we investigated the roles of MMP-2 and -9 on the H-ras-induced invasive phenotypes of MCF 10A human breast epithelial cells and HT 1080 human fibrosarcoma cells. We show that H-ras is able to induce or enhance a signaling pathway leading to the enhancement of an invasive phenotype in both MCF10A and HT1080 cells as determined by matrigel invasion assay. We then examined the effect of H-ras activation on the expression of MMP-2 and -9 by measuring enzymatic activities and mRNA levels. Our data clearly demonstrated that H-ras prominently induces expression of MMP-2 in MCF10A cells, while it efficiently up regulates MMP-9 in HT1080 cells. Taken together, these findings suggest that the correlation between ras-mediated invasiveness and enhanced expression of MMPs may be cell type-specific: MMP-9 is closely associated with the invasive phenotype induced by ras activation in fibrosarcoma cells, whereas MMP-2 is more likely associated with it in epithelial cells.

• The Journal of Korean Physical Therapy
• /
• 제23권1호
• /
• pp.59-66
• /
• 2011

Purpose: The purpose of this study was to investigate the effect of electroacupuncture on pain threshold and 5-HT synthesis in raphe magnus nucleus of dog brain stem following nociception induced by 1% carageenan. Method: Twenty male dogs were randomly divided into two groups, a control group and an electroacupuncture group with the following parameters: ST36, LI11 and SP9 with 120Hz and 0.5mA. The dogs were sacrificed at 30, 60, 90, 120, 150 and 180 min after nociception using 1% carageenan injection into the plantar side of the dog's left leg. The pain threshold was recorded by an Analgesia-meter. After making brain slice sections, the tissues were immunostained with 5-HT antisera (1:2,500). Results: The pain threshold for the electroacupuncture group was higher than for the control group. The number of 5-HT immunoreactive neurons in the electroacupuncture group was greater than that of the control group. Differences between the two groups regarding the number of 5-HT immunoreactive neurons at 30 min and 60 min (p<0.05), at 120 min and 150 min (p<0.01), and at 8 days (p<0.05) were significant. Conclusion: The increases in pain threshold and number of 5-HT immunoreactive neurons in the raphe magnus nucleus of dog brain stem showed that electroacupuncture can attenuate pain transduction through the descending tract.

• 한국식품영양학회지
• /
• 제35권6호
• /
• pp.464-472
• /
• 2022

The present study was designed to investigate the antiproliferative activity and molecular mechanisms of Bibimbap in HT-29 human colorectal adenocarcinoma cells. Bibimbap extract inhibited the proliferation of HT-29 cells by 50% at a concentration of 10.1±0.17 mg/mL for 48 h. The population of live cells decreased slightly, and the morphology changed with a reduction in cell volume (pyknosis) with Bibimbap. Treatment with 5 mg/mL of Bibimbap resulted in slight cell shrinkage. Furthermore, as the Bibimbap dose increased to 10 mg/mL, these characteristics were more evident, and HT-29 cells exhibited partial detachment by staining with the DNA-binding dye Hoechst 33342. Flow cytometric analysis by Annexin V and PI double staining showed that Bibimbap increased the levels of apoptosis. Analysis of the mechanism of these events showed that Bibimbap-treated cells exhibited a mitochondria-dependent apoptotic pathway through the modulation of caspase-3, caspase-8, caspase-9, and poly-ADP ribose polymerase, as well as Bax and Bcl-2 expression in dose- and time-dependent manners. Consequently, Bibimbap exerts a significant antiproliferative effect on HT-29 human colorectal adenocarcinoma cells.

• 미생물학회지
• /
• 제34권4호
• /
• pp.194-199
• /
• 1998

한강의 본류와 만나는 탄천과 중랑천에서 16S rDAN를 증폭하고 부분적인 염기서열 분석을 통하여 한강의 세균 다양성을 결정하였다. 총 27개의 클론을 분리하였으며 RFLP를 이용하여 7개의 group으로 나누었다. 탄천의 15개 클론은 4개의 group으로 나뉘어졌으며 가장 많은 클론을 포함하는 group(HT-1 클론)은 class Proteobacteria의 ${\delta}$-subdivision에 속하는 Acrobacter cryaerophilius와 높은 유사도를 보였으며, 다른 두 group(HT-6과 HT-9 클론)은 모두 clas Cytophagales에 속하였다. 중랑천의 12개의 클론은 3개의 group으로 나뉘어졌으며 가장 많은 클론을 보이는 group(HJ-1 클론)은 class Proteobacteria의 ${\alpha}$-subdivision에 속하는 Sphingomonas sp. 와 높은 유사도를 나타내었다. 전체적으로는 Proteobateria(alpha, beta and delta subdivision), Cytophagales와 Actinomycetales가 검출되었다.

• 대한약리학회지
• /
• 제2권1호
• /
• pp.71-82
• /
• 1966

The inhibitory effect of 5-hydroxytryptamine (5-HT) on the isolated intestinal strips of the tortoise (Amyda japonica), rat, rabbit and guinea pig was investigated. 1) The strips from the middle or lower part of the tortoise intestine responded with relaxation to 5-HT $(10^{-9}{\sim}10^{-5}g/ml)$, and the magnitude of the relaxation was proportional to the dose of 5-HT. The rectal part of the tortoise intestine, in contrast, showed contraction, the magnitude of which also was proportional to the dose of 5-HT. 2) Various blocking agents such as methysergide, morphine, tetracaine, nethalide, bretylium, hexamethonium, mecamylamine and chlorisondamine, showed no selective blocking activity on the relaxant effect of 5-HT on the tortoise intestine. The inhibitory effect of isoproterenol on the tortoise intestine, however, was selectively blocked by nethalide, and the stimulatory effect of 5-HT on the rectal part of the tortoise was blocked by methysergide. 3) In the presence of 5-HT, the stimulatory effect of DMPP on the tortoise intestine was remarkably attenuated, whereas that of acetylcholine and $BaCl_2$ was little affected. In the presence of isoproterenol, the stimulatory effect of acetylcholine and $BaCl_2$ were affected, but that of DMPP was little affected. 4) Large dose of 5-HT($10^{-4}$g/ml) produced inhibitory effect on the strips from the distal part of the isolated colon of the rat, rabbit and guinea pig, when the strips had been exposed to 5-HT($10^{-4}$g/ml), methysergide or phen`oxybenzamine. 5) Bretylium, as well as nethalide, abolished or remarkably reduced, in a few cases of the experiments, the inhibitory effect of the large dose of 5-HT on the distal part of the colon, whereas morphine did not affect it. 6) The ileal strips of the guinea pig also showed relaxation, as in the colonic strips, having been exposed to the large dose of 5-HT or phenoxybenzamine. This effect, however, was not obsered in the case of the rabbit ileum. 7) The property of the action-site of 5-HT in the tortoise intestine seemd to be different from the 5-HT receptors which have been revealed by several investigators. 8) Adrenergic component seemed to be participated in the inhibitory effect of 5-HT on the colon of the rat and rabbit.

• 미생물학회지
• /
• 제42권4호
• /
• pp.319-325
• /
• 2006

칡을 첨가하여 배양한 아가리쿠스버섯(Agaricus blaxei Murill)으로 부터 열수추출, gel filtration chromatography, DEAE ion exchange chromatography를 통하여 아가리쿠스 $\beta$-glucan (AG)을 추출하였다. 추출한 아가리쿠스 $\beta$-glucan에 Bacillus megaterium 유래의 endo-$\beta$-(1$\rightarrow$6)-glucanase를 처리하여 올리고당류(AO)를 얻었다. 이렇게 얻은 AG와 AO를 이용하여 HT-29 인체 대장암 세포에 대한 항암 활성을 조사한 결과, 암세포의 성장 억제 효과는 시료의 처리 농도에 의존적으로 증가하였으며, apoptosis assay에서 암세포의 apoptosis 유발이 농도에 의존적으로 증가되었다. 또한, 암세포의 세포 주기를 분석한 결과, apoptosis 발생을 뜻하는 G0 (sub-G1)기와 G1기의 비율이 증가한 반면 S기와 G2/M기는 대조군에 비해 감소되었다. 이러한 결과를 바탕으로 암세포의 apoptosis 증가에 대한 AO의 작용이 어떤 유전자와 연관이 있는지를 알아보기 위하여 cDNA microarray를 통해 유전자의 발현율을 검색한 결과, apoptosis의 내부 외부 경로에 영향을 주는 유전자(TNESE9, TNFRSF9, FADD, CASP8, BAD, CRADD, CASP9 등)의 발현이 증가되었고 세포 분열 주기의 진행과 관련된 유전자(CCND2와 CDK2)의 발현이 감소되었으며, 세포 분열 주기를 지체시키는 유전자 (CDKN2A)의 발현은 증가되었다. 또한, 사이토카인을 암호하는 유전자 (IL6, IL18, IL6R 등)와 tumor suppressor와 관련된 유전자 (CEACAM1, TP53BP2, IRF1및 PHB)의 발현이 2배 이상 증가된 것을 확인할 수 있었다. 따라서 HT-29 인체 대장암세포에 대한 AO의 성장 억제 작용은 G0/Gl기를 지체시켜 암세포 증식을 억제하고 apoptosis에 의해 암세포를 사멸시키는 항암 활성을 나타내는 것으로 확인되었으며, 특허 AO가 AG보다 현저한 활성을 보였다. 더 나아가 아가리쿠스 $\beta$-glucan (AG)과 올리고당류 (AO)는 항암 활성을 가진 대체 의약 소재로 개발될 수 있을 것으로 기대된다.

• 동의생리병리학회지
• /
• 제18권6호
• /
• pp.1608-1612
• /
• 2004

This study was conducted to investigate angiogenic inhibitory effect of Zingiberis Rhizoma methanol extract using ECV-304 cells and HT1080 fibrosarcoma cells. The viability of ECV-304 was 30% at 50㎍/㎖ of Zingiberis extract and that of HT1080 was 30% at 100㎍/㎖. Using the BrdU incorporation assay, Zingiberis inhibited the DNA synthesis of ECV-304 and HT1080 by 70% and 50% at 200㎍/㎖. In tube formation assay, at 10㎍/㎖ of Zingiberis, tube network began to degrade and at higher doses, it was completely destroyed. Zymography demonstrated that Zingiberis extract decreased MMP-9 at 10㎍/㎖ and higher doses remarkably inhibited the expression of MMP-9. These data indicate that Zingiberis Rhizoma has angiogenic inhibitory effects and shows the possibility of future anti-metastatic drug.

• BMB Reports
• /
• 제43권1호
• /
• pp.62-68
• /
• 2010

Matrix Metalloproteinases (MMPs) are a family of zinc-endopeptidases which can degrade extracellular matrix (ECM) components and play important roles in a variety of biological and pathological processes. 6,6'-bieckol isolated and characterized from an edible marine brown alga Ecklonia cava (EC), according to the comprehensive spectral analysis of MS and NMR data. Here the influence of 6,6'-bieckol on expressions of MMPs was examined by zymography and western blot analysis via human fibrosarcoma cell line (HT1080). It is shown that 6,6'-bieckol significantly down regulated the expressions of MMP-2 and -9 in dose-dependent manner. The influence of 6,6'-bieckol on the cell viability and cell behavior of HT1080 cells were also investigated, our dates shown that it suppressed the migration and 3D culture in HT1080 cells. Meanwhile, we explored several signal pathways which may contribute to this process, and found the suppressing of MMPs expressions in HT1080 cells might be due to the suppression of NF-${\kappa}B$ signal pathway.