• 한국양식학회지
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• 제6권2호
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• pp.107-123
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• 1993

조피볼락의 필수지방산인 n-3HUFA가 어체에 미치는 생화학적 변화를 알아보기 위하여, n-3HUFA 함량이 $0.0\~1.5\%$가 되도록 조정한 6종의 실험사료로 조피볼락 치어를 10주간 사육하여 혈액성상, 혈청성분 및 간세포 성상의 변화를 검토하였다. 혈액성상(RBC, Hb, Ht, MCV, MCHC 및 MCH)은 사료의 n-3HUFA 함량에 따라 유의적인 차이를 나타내지는 않았다. 혈청단백질 농도$(2.76\~3.63g/100ml)$ 및 혈당량$(36\~71mg/100ml)$은 n-3HUFA $0.9\%$ 사료까지는 증가하다가 그 이상에서는 일정한 값을 유지하였다. 혈중 total cholesterol 농도$(137\~86mg/100ml)$와 free cholesterol 농도$(84.2\~63.8mg/100ml)$는 n-3HUFA $1.2\%$까지 점차 감소하는 경향을 보였으며, 그 이상에서는 일정 하였다. 혈청 GPT $(159.7\~47.0 Karmen\; unit)$, GOT $(28.0\~11.1\;Karmen\;unit)$ 및 LDH $(3888\~1539\;W-unit)$도 모두 n-3HUFA $1.2\%$까지는 감소 경향을 보이다가 그 이후에는 일정한 값을 유지하였다. n-3HUFA 무첨가 사료를 먹은 실험어의 간조직은 n-3HUFA $1.5\%$ 첨가 사료를 먹은 실험어에 비해 인포색의 구조 붕괴와 부분적인 세포 융해가 관찰되었다. 간 cytosol의 LDH 활성과 microsomal membrane의 $Ca^{2+}-ATPase$ 활성도 변화되어 사료의 n-3HUFA가 증가할수록 증가한 경향을 보였다.

• Applied Microscopy
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• 제18권2호
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• pp.1-20
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• 1988

The species of the slug used in the experiment is Limax flavus L. For identifying the chemical characteristics of the epidermis, granules and mucus-producing cell of this animal is examined with methylene blue-basic fuchsin double stain and PAS-alcian blue reagent. For the ultrastructural research of the epidermal free surface, the epitheial cell and the parenchymal cell are used with scanning electron microscope and transmission elec-tron microscope respectively. I . Epidermal tissue The epidermal tissue of the slug is observed being divided into the dorsal and the ventral side(toot pad) respectively. 1) Dorsal epidermal tissue The dorsal epidermis of the slug is constituted with the simple columnar epithelium and the microvilli are compacted on the epidermal free surface. Two different types of the secretory granules of the neutral and the acid mucus are observed between the epithelial cells, and the neutral mucous granules are highest electron-dense but the acid mucous granules are observed to be electron-lucent. 2) Foot epidermal tissue The Foot epidermis is formed with the taller simple columnar epithelium than the dorsal epidermis and these cells have both a large number of the microvilli and a few number of the large villi. The secretory granules of three different types, which are acid, neutral and mixed mucous granule of two different types are observed between the epithelial cells. The neutral mucous granules are highest electron dense but the acid mucous granules are observed to be electron-lucent. II . Mucous granule-producing cell and mucus-producing cells Seven different types of the granules-producing cell and the mucus-producing cells are observed between the parenchyma. 1) A-type of acid mucous granule-producing cell The electron-lucent granules are largely occupied in the cytoplasm of these cells and then the granules are surrounded by irregular membrane. These electron-lucent granules exhibit alcianophilia with PAS-alcian blue reaction, so these granules are certified to be acid mucopolysaccharide. 2) B-type of acid mucus-producing cell The nucleus and the cytoplasm of these cells are pushed by the acid mucus of the electron-lucent toward the cell membrane. This mucus has been confirmed to be the acid mucopolysaccharide with PAS-alcian blue reagent. 3) A-type of neutral mucous granule-producing cell These cells contain the electron-dense round granules with approximately $1{\mu}m$ in diameter, which exhibit strongly PAS-positive reaction. These granules are confirmed to be the neutral mucoplysaccharide. 4) B-type of neutral mucous granule-producing cell These cells contain two different types of electron dense granules and electron-lucent granules; The former exhibits to be strongly PAS-positive and the latter to have alcianophilia reaction respectively. 5) C-type of neutral mucus-producing cell These cells are similar to the shape and the size of the B-type of mucus-producing cell but these two different types of cells are stained with reversing properties to each other. The mucus of the C-type cell that electron-lucent is largely occupied in the cytoplasm that exhibits strongly PAS-positive reaction. 6) D-type of neutral mucous granule-producing cell These cells contain round granules about $1{\mu}m$ in size which are observed to be medium electron-dense granules and those granules are stained brightly red with PAS-weak positive reaction. The granules are certified to be neutral mucopolysaccharide. 7) E-type of neutral mucous granule-producing cell These cells are similar to the shape and the size of the D-type of neutral mucous granule-producing cell. These cells contain a large number of granules with about $1{\mu}m$ in diameter showing electron-lucent and then granules are seen to be PAS-weak positive reaction. III. Parenchyma The clear cell and dark cell are found in the parenchyma of the Limax flavus L. 1) Clear cell These cells are round formed and the nucleus of the cells are larger than cytoplasm. These cells which have the electron-lucent cytosol possess poorly developed organelles. 2) Dark cell These cells are found to be dark cells due to high electron-density, which exhibit strongly methylene-blue reaction from double stain of methylene blue-basic fuchsin.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.918-922
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• 2001

In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

• Nutrition Research and Practice
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• 제8권4호
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• pp.377-385
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• 2014

BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.

• 한국식품과학회지
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• 제21권6호
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• pp.884-889
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• 1989

이유한 수컷 흰쥐를 혈장 콜레스테롤의 수준에 따라 미리 4군으로 분류한 다음 옥수수배아유, 돈지 및 달맞이꽃 종자유를 15% 함유하고 콜레스테롤이 0.5%씩 첨가된 실험식이로 사육하였다. 콜레스테롤을 첨가하지 않은 대조군의 식이유지 급원으로는 옥수수배아유를 사용하였다. 4주간의 사육실험 후 혈장 콜레스테롤과 적혈구막 및 대동맥의 지방산조성을 비교 분석하였다. 그 결과 달맞이꽃 종자유를 급여한 흰쥐는 다른 실험군에 비하여 혈장 콜레스테롤이 낮은 수준을 나타내었으며 옥수수배아유 섭취군에서 식이 콜레스테롤의 첨가 유무가 혈장 콜레스테롤의 수준에 큰 영향을 미치지는 못하였다. 그리고 상기 조직에서 달맞이꽃 종유의 섭취군에서 GLA의 대사산물인 DGLA와 AA의 함량이 증가하였다.

• 대한본초학회지
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• 제36권5호
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• pp.125-133
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• 2021

Objectives : Thymus quinquecistatus var. japonica (H.Hara) is a member of the genus Thymus of perennial aromatic herb, and it's designated as a natural monument of South Korea. It has traditionally been known to have protective or therapeutic effects on various human disease including cerebrovascular and neurological disease. Recently it was suggested that essential oil extracted from thyme has anti-fungal and anti-bacterial effect. The aim of this study is to investigate anti-inflammatory effect of Thymus quinquecistatus var. japonica in Raw 264.7 macrophage cell line. Methods : The cytotoxic effects of water and 70% ethanol extracts from Thymus quinquecistatus var. japonica, was tested using MTT assay. Inhibitory effects of the extracts to nitric oxide production and mRNA expression of inflammatory cytokines were examined by RT-PCR. Also, MitoSOX-red assay and JC-1 assay were performed to determine if the extracts can inhibit mitochondrial ROS accumulation and maintain mitochondrial membrane potential. Results : In LPS-induced inflammatory response, the extracts efficiently reduced nitric oxide NO production through inhibiting mRNA expression of iNOS enzyme. In addition, expression of the proinflammatory cytokines, IL-1𝛽 and IL-6, was also down-regulated by the extract treatments. Excessive accumulation of mitochondrial ROS induced by LPS was inhibited in the extract treated cells, which finally protected mitochondrial membrane potential. Conclusions : These results showed that water and 70% ethanol extracts from Thymus quinquecistatus var. japonica have anti-inflammatory effect through down regulation of IL-1𝛽, IL-6, and iNOS, and also have antioxidative effect against mitochondrial ROS accumulation that promote inflammatory response.

• Journal of Microbiology and Biotechnology
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• 제31권2호
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• pp.217-225
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• 2021

This study aimed to investigate the neuroprotective effects of 1-methoxylespeflorin G11 (MLG), a pterocarpan, against glutamate-induced neurotoxicity in neuronal HT22 hippocampal cells. The protective effects of MLG were evaluated using MTT assay and microscopic analysis. The extent of apoptosis was studied using flow cytometric analysis performed on the damaged cells probed with annexin V/propidium iodide. Moreover, mitochondrial reactive oxygen species (ROS) were assessed using flow cytometry through MitoSOXTM Red staining. To determine mitochondrial membrane potential, staining with tetramethylrhodamine and JC-1 was performed followed by flow cytometry. The results demonstrated that MLG attenuates glutamate-induced apoptosis in HT22 cells by inhibiting intracellular ROS generation and mitochondrial dysfunction. Additionally, MLG prevented glutamate-induced apoptotic pathway in HT22 cells through upregulation of Bcl-2 and downregulation of cleaved PARP-1, AIF, and phosphorylated MAPK cascades. In addition, MLG treatment induced HO-1 expression in HT22 cells. These results suggested that MLG exhibits neuroprotective effects against glutamate-induced neurotoxicity in neuronal HT22 cells by inhibiting oxidative stress and apoptosis.

• Pediatric Infection and Vaccine
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• 제16권2호
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• pp.215-219
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• 2009

5-14세에 호발하는 마이코플라즈마 폐렴은 20-25%에서 폐외 증상을 일으키는 것으로 알려져 있으며 이 중 혈액계 질환에는 용혈성 빈혈, 혈소판 감소증, 혈구포식세포 증식증(hemophagocytosis) 등이 있다. 마이코플라즈마 폐렴에서의 냉항체에 의한 자가면역성 용혈로 용혈성 빈혈이 발생할 수 있으며, 이 때 항체의 역가와 용혈의 정도에 상관관계가 있을 수 있다. 저자들은 마이코플라즈마 폐렴에 의해 용혈성 빈혈이 발현된 유전성 구상 적혈구증 1례를 경험하였기에 보고하는 바이다.

• 미생물학회지
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• 제33권3호
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• pp.170-174
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• 1997

강한 항진균 활성을 나타내며, 낮은 적혈구 용혈활성을 갖는 새로운 합성 펩티드를 얻기 위하여 cecropin A(1-8)-magainin 2(1-12) 및 이들의 유사체들을 설계하고 이들을 고상법에 의하여 합성하였다. 합성 펩티드의 항진균 활성은 Trichosporon beigelii에 대한 성장억제능에 의하여 측정하였으며, 세포독성은 인간의 적혈구 세포에 대한 용혈활성에 의하여 측정하였다. 펩티드의 양쪽 친매성의 증가는 항진균활성보다는 적혈구 용혈황성에 큰 영향을 미쳤다. Cecropin A(1-8)-magainin 2(1-12)의 12번 아미노산이 Lys을 Ala으로 치환시킨 유사체 펩티드(A2)는 가장 강한 항진균활성(minimum inhibitory concentration : 2.5.$\mu$g/ml)을 나타내며 비교적 낮은 적혈구 용혈활성(200.$\mu$g/ml의 펩티드 농도에서 0.5% hemolysis를 나타냄)을 나타내었다. 따라서 A2의 펩티드는 세포독성을 갖지 않으며 강한 항진균 활성을 가지는 모델로서 유용하게 사용될 것이다.

• Toxicological Research
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• 제17권
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• pp.117-125
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• 2001

Envenoming by Russells viper causes a broad spectrum of renal impairment. Renal failure is an important complication in patients bitten by Russells viper. Experimental work in animals and in vitro has elucidated pathophysiological mechanisms that contribute to life threatening complications and have suggested possibilities for therapeutic intervention. The evidence in experimental animals regarding mechanisms of venom action in relation to changes in either extrarenal or intrarenal factors is presented. The cardiovascular system and renal hemodynamics are affected by venom. Reductions of renal function including renal hemodynamics are associated directly with changes in general circulation during envenomation. Possible endogenous mechanisms for releasing the hormone inducing renal vasoconstriction after envenomation are evident. Hormonal factor such as the catecholamine, prostaglandin and renin angiotensin systems induce these changes. Direct nephrotoxicity of venom action is studied in the isolated per-fused kidney. Characteristic polarization of the cell membrane, changes of mitochondrial activity and Na-K ATPase in renal tubular cells are observed. Changes in renal function and the cardiovascular system are observed of ter envenomation and are reversed by the administration of Russells viper antivenom (purified equine immunoglobulin, $Fab_2$ fragment). The neutralizing effects are more efficient when the intravenous injection of antivenom is given within 30 min after the envenomation.