• Title/Summary/Keyword: Vacuoles

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Interactions between secreted GRA proteins and host cell proteins across the parasitophorous vacuolar membrane in the parasitism of Toxoplasma gondii

  • Ahn, Hye-Jin;Kim, Sehra;Kim, Hee-Eun;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.44 no.4 s.140
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    • pp.303-312
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    • 2006
  • Interactions between GRA proteins of dense granules in Toxoplasma gondii and host cell proteins were analyzed by yeast two-hybrid technique. The cMyc-GRA fusion proteins expressed from pGBKT7 plasmid in Y187 yeast were bound to host cell proteins from pGADT7-Rec-HeLa cDNA library transformed to AH109 yeast by mating method. By the selection procedures, a total of 939 colonies of the SD/-AHLT culture, 348 colonies of the $X-\alpha-gal$ positive and PCR, 157 colonies of the $X-\beta-gal$ assay were chosen for sequencing the cDNA and finally 90 colonies containing ORF were selected to analyze the interactions. GRA proteins interacted with a variety of host cell proteins such as enzymes, structural and functional proteins of organellar proteins of broad spectrum. Several specific bindings of each GRA protein to host proteins were discussed presumptively the role of GRA proteins after secreting into the parasitophorous vacuoles (PV) and the PV membrane in the parasitism of this parasite.

Electron Microscopic Study on the Spermiogenesis of Cipangopaludina chinensis malleata (Reeve) (논우렁이[Cipangopaludina chinensis malleata (Reeve)]의 精蟲形成 過程에 관한 電子顯微鏡的 硏究)

  • Kim, Ju-Hee;Park, Won-Chul
    • The Korean Journal of Zoology
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    • v.29 no.2
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    • pp.121-140
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    • 1986
  • The spermiogenesis of Cipangopaludina chinensis malleata began with the changes of nucleus and cytoplasm. The chromatin in the nucleus began to stack and condense around perinuclear envelope. Axonema (doublet microtubules of 9+2) appeared in the cytoplasm. After this process, the cytoplasm was protruded and the bell-shaped nucleus was located on protruding part of it. The electron dense masses were distributed throughout cytoplasm and lysed or secreted by exocytosis of lysosomal vacuoles. Especially, some mitochondria were migrated by the doublet microtubules of axonema toward nucleus. The axoneme was enclosed by electron dense materials after exocytosis of unnecessary materials for the tail formation. The electron dense masses were released and migrated into the each part through microtubules or axonemal doublet microtubules as the granular particles. These granular particles were containing glycogen. Ultimately, the condensed head developed into helical and neck region into cylindrical shape respectively. The mitochondria which have regular lamellar layers at cross axis became to middle piece, and then spermatozoon was completely matured. Thus, these phenomena showed special processes in spermiogenesis, those were as follows; chromatin in the head was condensed, the head was changed into helical shape, and cytoplasmic materials are migrated and transferred into the each part in the tail by mitochondria and microtubules.

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The inhibitory effects of Soongijeseub-bang(順氣除濕方) on the obese-mice caused by high-fat diet (순기제습방(順氣除濕方)이 고지방식이(高脂肪食餌)로 유발된 비만(肥滿) 백서(白鼠)에 미치는 영향)

  • Lee, Woo-Yeol;Shin, Hyun-Kyoo
    • Korean Journal of Oriental Medicine
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    • v.14 no.1
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    • pp.161-171
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    • 2008
  • Objective : In order to investigate the effects of Soongijeseub-bang(here in after referred to SJB) on the obese gene and obese inhibitory, C57BL/6 mice were induced by high fat diet Methods : C57BL/6 mice were divided into three groups(normal, high fat diet with control, high fat diet with SJB extract) and fed for 15 weeks. And measured body weight change, the weight change of the adipocytes in body, the levels of ALT, AST, creatinine, total cholesterol, LDL-cholesterol, HDL-cholesterol, triglyceride, NEFA(non-esterified fat acid), glucose in serum and the expression of ${\beta}3AR$, leptin, and TNF- ${\alpha}$ gene in primary adipocytes and adipocytes tissue. Results : The following results were obtained in this study. 1. SJB 500 mg/kg extract group showed significant decrease in final weight. 2. SJB 500 mg/kg extract group showed significant decrease in the amount of adipocyte in weight. 3. All experimental group showed significant decrease in ALT and AST levels. 4. All experimetal group did not show significant decrease in glucose and creatinine levels. 5. All experimental group showed significant decrease in total cholesterol, LDL-cholesterol, triglyceride and NEFA levels. HDL-cholesterol were increase significantly in SJB 500 mg/kg extract group. 6. SJB 500 mg/kg extract group showed significant decrease in leptin levels. 7. All experimental group showed significantly increased expressions of fJ 3AR in primary adipose cell and 3T3-Ll cell, and those of leptin in primary adipose cell and 3T3-Ll cell were decreased significantly. SJB $100{\mu}l/ml$ extract group showed significant decrease in the expression of TNF-${\alpha}$. 8. SJB 500 mg/kg extract group showed decrease in the size of adipocyte in adipocytes tissue. 9. All experimental group showed significant decrease in adipose vacuoles in liver tissue.

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Fine Needle Aspipration Cytology of Metastatic Pulmonary Seminoma - A Cese Report - (세침흡인 세포검사로 진단된 폐에 전이한 정상피종 - 1예 보고 -)

  • Jeong, Hwa-Sook;Lee, Geon-Kook;Kim, Wun-Jae;Earm, Jae-Ho;Song, Hyung-Geun
    • The Korean Journal of Cytopathology
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    • v.7 no.1
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    • pp.97-102
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    • 1996
  • Fine needle aspiration cytology of a pulmonary mass was performed on a 51-year-old man who had a left testicular mass. Cytologic features were composed of a homogeneous population of malignant cells associated with a background of foamy and lacelike material. The cellular features were characterized by monomorphous cell proliferation of relatively regular large cells, generally isolated or grouped. Occasionally, fine blanching stroma with large tumor cells and scanty lymphocytes were noted. The tumor cells had a round, regular nucleus, prominent round nucleoli, and a thin rim of cytoplasm containing large vacuoles or lacunae filled with glycogen. The fine needle aspiration cytologic diagnosis was highly consistent with metastatic seminoma from testis and less likely primary or other metastatic carcinoma. The diagnosis of resected testicular mass was classic seminoma. Despite the fact that cytopathologists were not familial with diagnosis of seminoma due to clinician's lack of interest in fine needle aspiration cytology of germ cell tumors including seminoma, it appears that a diagnosis of this tumor should not be problematic in cytologic material if specific histologic criteria are applied.

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The Inhibitory Effects of Bimanbang-1(肥滿1號方) on the Obese-mouse Fed High-fat Diet (비만1호방(肥滿1號方)이 고지방식이(高脂肪食餌)로 비만(肥滿) 유도(誘導)된 백서(白鼠)에 미치는 영향)

  • Shin, Hong-Jung;Yoon, Il-Ji
    • The Journal of Korean Medicine
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    • v.29 no.2
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    • pp.116-132
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    • 2008
  • Objective: This study investigated the effects of Bimanbang-1 (肥滿1號方: here in after referred to BMB-1) on the obese gene and obese inhibitory. Methods: C57BL/6 mice were induced by high fat diet. Mice were divided into three groups (normal, high fat diet with control, high fat diet with BMB-1 extract) and fed for 15 weeks. Items of this experimental study are as follows: body weight change, final body weight, the weight change of the adipocytes in body, the level change of LFT, NEFA and creatinine, the expression of ${\beta}3AR$ and leptin gene in primary adipocytes, the production change of leptin in primary adipocytes, the expression of ${\beta}3AR$, leptin and $TNF-{\alpha}$ in adipocytes tissue. Result: 1. All experimental groups showedthat the weight change decreased considerably and the high density group showedthat the final weight decreased considerably. 2. The high density group showed that the amount of the adipocyte in weight decreased considerably. 3. All experimental groups showedthat the amount of ALT decreased considerably, and AST decreased in the high density group. However, the amount of creatinine and glucose did not increase considerably. 4. All experimental groups showed that the amount of total cholesterol, LDL-cholesterol, triglyceride, and NEFA decreased, and HDL-cholesterol increased considerably. 5. The high density groups showedthat the amount of leptin decreased considerably. 6. All experimental groups showed that the revelation of ${\beta}3AR$ in primary adipose cell and 3T3-L1 cell increased considerably, and that the revelation of leptin and $TNF-{\alpha}$ in primary adipose cellsand 3T3-L1 cells decreased considerably. 7. All experimental groups showed that the size of adipocyte in adipocytes tissue decreased. 8. All experimental groups showed that the adipose vacuoles in liver tissue decreased considerably. Conclusion: The findings suggest that Bimanbang-1 causes weight loss and histological change, thus it may be effective to treat obesity.

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Oleanolic acid induced autophagic cell death in hepatocellular carcinoma cells via PI3K/Akt/mTOR and ROS-dependent pathway

  • Shi, Yang;Song, Qingwei;Hu, Dianhe;Zhuang, Xiaohu;Yu, Shengcai;Teng, Dacai
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.3
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    • pp.237-243
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    • 2016
  • Oleanolic acid (OA) has a wide variety of bioactivities such as hepatoprotective, anti-inflammatory and anti-cancer activity and is used for medicinal purposes in many Asian countries. In the present study, the effect of OA on induction of autophagy in human hepatocellular carcinoma HepG2 and SMC7721 cells and the related mechanisms were investigated. MTT assay showed that OA significantly inhibited HepG2 and SMC7721 cells growth. OA treatment enhanced formation of autophagic vacuoles as revealed by monodansylcadaverine (MDC) staining. At the same time, increasing punctuate distribution of microtubule-associated protein 1 light chain 3 (LC3) and an increasing ratio of LC3-II to LC3-I were also triggered by OA incubation. In addition, OA-induced cell death was significantly inhibited by autophagy inhibitors 3-methyladenine (3-MA) and chloroquine (CQ) pretreatment. And we found out that OA can suppress the PI3K/Akt1/mTOR signaling pathway. Furthermore, our data suggested that OA-triggered autophagy was ROS-dependent as demonstrated by elevated cellular ROS levels by OA treatment. When ROS was cleared by N-acetylcysteine (NAC), OA-induced LC3-II convertsion and cell death were all reversed. Taken together, our results suggest that OA exerts anticancer effect via autophagic cell death in hepatocellular carcinoma.

The treatment effect of novel hGHRH homodimer to male infertility hamster

  • Zhang, Xu-Dong;Guo, Xiao-Yuan;Tang, Jing-Xuan;Yue, Lin-Na;Zhang, Juan-Hui;Liu, Tao;Dong, Yu-Xia;Tang, Song-Shan
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.6
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    • pp.637-647
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    • 2018
  • Extra-hypothalamic growth hormone-releasing hormone (GHRH) plays an important role in reproduction. To study the treatment effect of Grin (a novel hGHRH homodimer), the infertility models of 85 male Chinese hamsters were established by intraperitoneally injecting 20 mg/kg of cyclophosphamide once in a week for 5 weeks and the treatment with Grin or human menopausal gonadotropin (hMG) as positive control was evaluated by performing a 3-week mating experiment. 2-8 mg/kg of Grin and 200 U/kg of hMG showed similar effect and different pathological characteristics. Compared to the single cyclophosphamide group (0%), the pregnancy rates (H-, M-, L-Grin 26.7, 30.8, 31.3%, and hMG 31.3%) showed significant difference, but there was no difference between the hMG and Grin groups. The single cyclophosphamide group presented loose tubules with pathologic vacuoles and significant TUNEL positive cells. Grin induced less weight of body or testis, compactly aligned tubules with little intra-lumens, whereas hMG caused more weight of body or testis, enlarging tubules with annular clearance. Grin presented a dose-dependent manner or cell differentiation-dependentincrease in testicular GHRH receptor, and did not impact the levels of blood and testicular GH, testosterone. Grin promotes fertility by proliferating and differentiating primitive cells through up-regulating testicular GHRH receptor without triggering GH secretion, which might solve the etiology of oligoasthenozoospermia.

Diethylnitrosamine-induced hepatic tumorigenesis in rats 2. Histological findings about apoptosis and proliferation of tumor cells (Diethylnitrosamine을 투여한 rat 간장의 tumorigenesis에 관하여 2. 종양세포의 apoptosis와 증식에 관한 조직학적 소견)

  • Kwak, Soo-dong;Kang, Chung-boo;Koh, Phil-ok
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.386-393
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    • 1998
  • The study was performed to investigate the histological findings and the appearances of positive cells by immunohistochemical methods using proliferating cell nuclear antigen (PCNA) antibody and apoptotic kit in diethylnitrosamine (DEN) -induced rat liver cancer model. Forty four male rats (Sprague Dawley), initially 5 to 6 weeks of age and 120 to 150gm in body weight were continuously were given with water containing 0.01% DEN for 13 weeks and 3~6 rats per week were randomly sacrified at intervals of a week from 8 weeks to 17 weeks. The interlobular connective tissues in the rat livers were proliferated at early 8 weeks. The vaccuolated or fatty degenerated liver cells were focally distributed and then widely distributed with the passage of weeks and the liver cells with large vacuoles tended to be crowded in focal areas, and the liver cells in some lobules were transformed into small or eosinophilic polyhedral large cells. The hepatocellular carcinoma and the cholangiocarcinoma were simultaneously developed in same liver and tended to be markedly developed after 12 weeks but the development of carcinoma in some livers at same week were less or more advanced as 3~5 week intervals. The regions with more number of positive cells by PCNA antibody or apoptotic kits in livers were ranked as following order ; small hepatocellular carcinoma regions, cholangiocarcinoma regions, trabecular or acinar type carcinoma regions, and large liver cell regions. The numbers of the positive cells by PCNA antibody were more numerous than those by apoptotic kit. So these findings suggested that the volumes and weights of the livers were increasing by more many proliferating of carcinoma cells on the above ordered regions.

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Colonial Cyanobacteria, Microcystis Cell Density Variations using Ultrasonic Treatment (초음파 처리 조건에 따른 집락형 유해남조류 Microcystis 세포수 변화 연구)

  • Lee, Hae-Jin;Park, Hae-Kyung;Heo, Jun;Lee, Hyeon-Je;Hong, Dong-Gyun
    • Journal of Korean Society on Water Environment
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    • v.34 no.2
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    • pp.210-215
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    • 2018
  • It is difficult to count colonial cyanobacteria Microcystis cells since the thickness of colonies is constrained by amorphous mucilage, making it impossible to estimate the number of cells. Disaggregation of Microcystis colonies into single cell is needed to improve the accuracy and precision of cell density estimation of naturally collected samples. Uultrasonic treatment method is commonly used owing to the simplicity and immediacy of the procedure. However, amplitude, frequency, and duration of ultrasonic treatment also cause cell loss during the experiment. Optimal ultrasonic treatment has not been standardized yet. Therefore, the objective of this study was to investigate optimal ultrasonic treatment by analyzing cell density and colony numbers. We collected colonial Microcystis from Changnyeong-Haman weir area in Nakdong River during harmful algal boom period from September to October in 2017. Ultrasonic treatment method was applied to disrupt colonies into single cells to enumerate cell density. Among treatment conditions, results from continuously treated for 100 seconds were found to be the optimum to reduce colonies to a suspension of single cell without cell losses under high and low density of Microcystis cells. Lugol iodine fixed cells followed by sonication showed less negative impact of cell damage within the optimal treatment time (100 seconds). Furthermore, disaggregated cells treated by sonication enables microscopic observation more easily since gas vacuoles were collapsed to facilitate sedimentation of cells under the counting chamber for quantitative enumeration of buoyant Microcystis cells.

ULTRASTRUCTURAL STUDY OF THE FIBROBLAST REMODELING IN THE RADIATION- IMPAIRED WOUND HEALING IN THE TONGUE OF THE RAT (방사선조사가 설의 개방성창상치유에서 섬유아세포의 재형성에 미치는 영향에 관한 연구)

  • Lee Jin-Koo;Choi Soon-Chul;Park Tae-Won;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.27 no.1
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    • pp.141-165
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    • 1997
  • Radiation-impaired wound healing in animal experiments was believed to be an another logical experimental model to understand the wound healing mechanism in patients. The purpose of this study was to reveal the block point which would result in impaired healing. Twenty four rats(Sprague-Dawley strains) were divided into two groups according to the time interval between irradiation and wounding. Group I, observing the healing effect on the 1st day and Group II are the healing effects on the 7th days after irradiation to the wound of the rat tongue. Experimental animals were sacrificed 3, 6, 12, and 24 hours after wounding. The specimens were examined by the light microscope and transmission electron microscope. The following results were obtained 1. Fibroblasts in both groups showed degenerative changes which were dilated mitochondria and rER, reduced microorganelle, vacuoles and little cytoplasmic process. 2. Average length between bands and Quantity of the newly produced collagen fibers around fibroblasts remained unchanged against control group. 3. The severity of degenerative change of the fibroblast and impairment of wound healing including shortening of the thickness of collagen fibers were more severe in the group II than in the group I.

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