• Title/Summary/Keyword: 2-cell fusion

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Serodiagnosis of human paragonimiasis by ELISA-inhibition test using monoclonal antibodies (단클론항체를 이용한 폐흡충증의 면역진단)

  • Yong, Tae-Sun;Seo, Jang-Hun;Yeo, In-Seok
    • Parasites, Hosts and Diseases
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    • v.31 no.2
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    • pp.141-148
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    • 1993
  • ELISA-inhibition test using Paragonimus westermani specific monoclonal antibody (Mab) was investigated to improve the diagnostic specificity of paragonimiasis. By cell fusion, one hybridoma clone secreting un-n westemanl specific Mab was selected (Pwa-14), which reacted on bands of 28 kDa, 42.5 kDa, 89 kDa and 120.5 kDa. IFA showed Pwa-14 was located at the vitelline follicles. By micro-ELISA, 100% of 22 paragonimiasis cases were found positive, but 5 of 40 clonorchlasls cases (12.5%),3 of 26 cystlcercosis cases (7.7%) showed false positive. None of 10 sparganosis patients or 28 normal controls reacted positively. On the other hand, by ELISA-Inhibition test using a R westermcni specific Mab, 100% of patagonimlasls cases were found positive, and there were no positive in cysticercosis, sparganosis cases or normal controls, except 2 (5.0%) false-positive sera of 40 clonorchiasis cases. The ELISA-Inhlbltlon test using a Mab showed higher specificity in comparison with macro-ELISA for serodlgnosis of human paragonimlasis.

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Ginsenoside Rg3 ameliorates myocardial glucose metabolism and insulin resistance via activating the AMPK signaling pathway

  • Ni, Jingyu;Liu, Zhihao;Jiang, Miaomiao;Li, Lan;Deng, Jie;Wang, Xiaodan;Su, Jing;Zhu, Yan;He, Feng;Mao, Jingyuan;Gao, Xiumei;Fan, Guanwei
    • Journal of Ginseng Research
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    • v.46 no.2
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    • pp.235-247
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    • 2022
  • Background: Ginsenoside Rg3 is one of the main active ingredients in ginseng. Here, we aimed to confirm its protective effect on the heart function in transverse aortic coarctation (TAC)-induced heart failure mice and explore the potential molecular mechanisms involved. Methods: The effects of ginsenoside Rg3 on heart and mitochondrial function were investigated by treating TAC-induced heart failure in mice. The mechanism of ginsenoside Rg3 for improving heart and mitochondrial function in mice with heart failure was predicted through integrative analysis of the proteome and plasma metabolome. Glucose uptake and myocardial insulin sensitivity were evaluated using micro-positron emission tomography. The effect of ginsenoside Rg3 on myocardial insulin sensitivity was clarified by combining in vivo animal experiments and in vitro cell experiments. Results: Treatment of TAC-induced mouse models with ginsenoside Rg3 significantly improved heart function and protected mitochondrial structure and function. Fusion of metabolomics, proteomics, and targeted metabolomics data showed that Rg3 regulated the glycolysis process, and Rg3 not only regulated glucose uptake but also improve myocardial insulin resistance. The molecular mechanism of ginsenoside Rg3 regulation of glucose metabolism was determined by exploring the interaction pathways of AMPK, insulin resistance, and glucose metabolism. The effect of ginsenoside Rg3 on the promotion of glucose uptake in IR-H9c2 cells by AMPK activation was dependent on the insulin signaling pathway. Conclusions: Ginsenoside Rg3 modulates glucose metabolism and significantly ameliorates insulin resistance through activation of the AMPK pathway.

Analysis of Growth and Hematologic Characteristics of Endangered Korean Native Cattle (멸종위기의 백한우(체세포복제 포함)의 성장 및 혈액학적 특성 분석)

  • Kim, Hyun;Choe, Changyong;Seong, Hwan-Hoo
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.175-181
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    • 2015
  • The objective of this study was to monitor health conditions of genetically identical somatic cells cloned Korean white cattle, endangered indigenous cattle (EIC) and indigenous cattle (IC) by analysis of hematologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and one live cattle were delivered by caesarean section. The cloned Korean white cattle were genetically identical to the nuclear donor cattle. As a result, the mean values of RBC and platelet of cloned cattle and white cattle were significantly decreased by age (P<0.05). The mean values of RBC, HCT, MCV and MCHC between cloned cattle and IC of the same age (1~2 years) showed the statistical significance (P<0.05). Also, in the WBC of Korean white cattle, the estimated values were decreased according to the age from $12.0{\times}10^3/{\mu}l$ under 1 year to $11.0{\times}10^3/{\mu}l$ over 1 years respectively. Although clone-cattle had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned cattle were not different when compared to reference range. This study suggests that cloned Korean white cattle derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological parameters. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

Reversibility and safety of KISS1 metastasis suppressor gene vaccine in immunocastration of ram lambs

  • Han, Yan-Guo;Liu, Gui-Qiong;Jiang, Xun-Ping;Xiang, Xing-Long;Huang, Yong-Fu;Nie, Bin;Zhao, Jia-Yu;Nabeel, Ijaz;Tesema, Birhanu
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.6
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    • pp.835-841
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    • 2018
  • Objective: The aim of this study was to investigate the reversibility and safety of KISS1 metastasis suppressor (KISS1) gene vaccine in immunocastration. Methods: Six eight-week old ram lambs were randomly divided into vaccinated and control groups. The vaccine (1 mg/ram lamb) was injected at weeks 0, 3, and 6 of the study. Blood samples were collected from the jugular vein before primary immunization and at weeks 2, 4, 6, 10, 14, 22, and 30 after primary immunization. All ram lambs were slaughtered at 38 weeks of age, and samples were collected. Results: The specific anti-KISS1 antibody titers in vaccinated animals were significantly higher and the serum testosterone level was significantly lower than those in the control groups from week 4 to 14 after primary immunization (p<0.05). No significant difference was observed at weeks 22 and 30 after the primary immunization. Similar results were also found for scrotal circumference, testicular weight, length, breadth, and spermatogenesis in seminiferous tubules in week 30 after primary immunization. KS (KISS1-hepatitis B surface antigen S) fusion fragment of KISS1 gene vaccine was not detected in host cell genomic DNA of 9 tissues of the vaccinated ram lambs by polymerase chain reaction. Conclusion: The effects of KISS1 gene vaccine in immunocastration were reversible and no integration events were recorded.

Pathogenesis on enteritis induced by Cryptosporidium parvum alone and combined with porcine rotavirus in piglets (Cryptosporidium parvum 단독 및 돼지 rotavirus와 혼합 감염시킨 자돈 장염의 병원성)

  • Han, Dong-un;Kang, Mun-il;Park, Nam-yong;Wee, Sung-hwan
    • Korean Journal of Veterinary Research
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    • v.35 no.1
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    • pp.149-158
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    • 1995
  • The purpose of the present study was to understand the pathogenesis of infections in piglets inoculated with C parvum isolated from mice alone and combined with porcine rotavirus (S-80). Thirteen 10-day piglets were divided in four groups; Three, A group, were only given by C parvum. Four, B group, were orally administrated with firstly porcine rotavirus and then C patvum. Three, C group, were orally inoculated with porcine rotavirus alone. The rest, D group, were used as controls. During the experiment, there were daily recorded clinical signs including diarrhea to each pig. According to the periodic intervals for necropsy, all pigs were sacrificed from 4 to 12 days after the final inoculation of C parvum. Location and distribution of two pathogens, C parvum and rotavirus, in the intestinal mucosa of piglets tested were examined by pathological and immunohistological means. In addition, parasitological test using the feces of piglets was applied for the detection of cryptosporidial oocysts as well. A group showed diarrhea from 4 to 6 days post-inoculation(PI) and also discharged C parvum oocysts in feces during the day 4 to 7 PI. In tissue sections of jejunum and ileum, cryptosporidial oocysts were observed a few on the top of villi with slightly fusion. B group represented sign of diarrhea and discharge of oocysts from 2 to 11 days PI. There were some cryptosporidial oocysts both in the jejunal lumen and in the lumen of mucosal glands. As progressed, oocysts were most commonly distributed on the tip of villi of jejunum. Histopathologically there were also mild to moderately fused, attenuated focal desquamated, congested villi and mononuclear cell infiltration of varying degrees in the lamina propria of small intestine and colon at the day 4 and 7 PI. C group showed slightly to mildly attenuated and fused top of villi and mildly mucosal congestion. D group as controls was grossly and histopathologically normal in all parts of intestine. The present results indicate that the piglets inoculated with C parvum only are certainly milder in pathogenesis including duration of clinical course and severity of lesion than those in piglets concurrently infected with porcine rotavirus and C parvum. Also the strain (VRI-CN91) of C parvum used in the study has very low pathogenicity to occur enteritis of piglets.

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Comparison of Inflammatory Markers Changes in Patients Who Used Postoperative Prophylactic Antibiotics within 24 Hours after Spine Surgery and 5 Days after Spine Surgery

  • Youn, Gun;Choi, Man Kyu;Kim, Sung Bum
    • Journal of Korean Neurosurgical Society
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    • v.65 no.6
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    • pp.834-840
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    • 2022
  • Objective : C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR), and white blood cell (WBC) count are inflammatory markers used to evaluate postoperative infections. Although these markers are non-specific, understanding their normal kinetics after surgery may be helpful in the early detection of postoperative infections. To compliment the recent trend of reducing the duration of antibiotic use, this retrospective study investigated the inflammatory markers of patients who had received antibiotics within 24 hours after surgery according to the Health Insurance Review & Assessment Service guidelines and compared them with those of patients who had received antibiotics for 5 days, which was proven to be non-infectious. Methods : We enrolled 74 patients, divided into two groups. Patients underwent posterior lumbar interbody fusion (PLIF) at a single institution between 2019 and 2020. Group A included 37 patients who received antibiotics within 24 hours after the PLIF procedure, and group B comprised 37 patients who had used antibiotics for 5 days. A 1 : 1 nearest-neighbor propensity-matched analysis was used. The clinical variables included age, sex, medical history, body mass index, estimated blood loss, and operation time. Laboratory data included CRP, ESR, and WBC, which were measured preoperatively and on postoperative days (POD) 1, 3, 5, and 7. Results : CRP dynamics tended to decrease after peaking on POD 3, with a similar trend in both groups. The average CRP level in group B was slightly higher than that in group A; however, the difference was not statistically significant. Multiple linear regression analysis revealed operation time, number of fused levels, and estimated blood loss as significant predictors of a greater CRP peak value (r2=0.473, p<0.001) in patients. No trend (a tendency to decrease from the peak value) could be determined for ESR and WBC count on POD 7. Conclusion : Although slight differences were observed in numerical values and kinetics, sequential changes in inflammatory markers according to the duration of antibiotic administration showed similar patterns. Knowledge of CRP kinetics allows the assessment of the degree of difference between the clinical and expected values.

Survival, Physiological Responses, and Histological Changes in Korean Rockfish (Sebastes schlegelii) Exposed to Artificial Increase of Water Temperature (인위적 수온 상승에 노출된 조피볼락(Sebastes schlegelii) 치어의 생존율 및 스트레스 반응)

  • Young Guk Jin;Hyun Woo Gil;Dae-Jung Kim;Hyungkyu Hwang;Hyo-Won Kim
    • Korean Journal of Ichthyology
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    • v.35 no.4
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    • pp.236-243
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    • 2023
  • This study aimed to investigate the survival rates, hematologic responses, and histological responses of juvenile Korean rockfish (Sebastes schlegelii) exposed to artificial increase of water temperature. The water temperature was incrementally raised from the initial 23℃ to 26℃, 28℃, 30℃, and 31℃, with a 1℃ increase every 24 hours. The fish were exposed to each water temperature setting for a period of seven days. No mortality was observed at 26℃ and 28℃. However, at 30℃, mortality began on the 4th day of exposure, with an overall survival rate of 1.5% at the end of the seventh day. At 31℃, mortality occurred as early as the first day of exposure, and all fish had perished by the second day. Plasma cortisol and glucose concentrations increased as water temperature rose, with a significant decrease observed at 31℃. No significant difference in plasma GPT concentration was observed across the various experimental temperatures. In contrast, plasma GOT concentration significantly increased at 31℃. Histological examination revealed that both the liver and gills exhibited normal histology at the initial temperature of 23℃ and at 26℃. However, at 28℃ hepatocellular hypertrophy and gill lamellar epithelial hyperplasia and epithelial cell lifting were observed. At 30℃, hepatocellular condensation and gill lamellar fusion were noted. Finally, at 31℃, severe histological changes were observed, including hepatocellular necrosis, liver congestion, and gill filament necrosis.

THE EFFECT OF BONE MORPHOGENETIC PROTEIN 2(BMP2) ON THE GROWTH OF CRANIAL BONE AND EARLY MORPHOGENESIS OF THE CRANIAL SUTURE (Bone Morphogenetic Protein 2 가 두개골 성장 및 두개봉합부의 초기형태발생에 미치는 영향)

  • Jung, Hae-Kyung;Park, Mi-Hyun;Ryoo, Hyun-Mo;Nam, Soon-Hyeun;Kim, Young-Jin;Kim, Hyun-Jung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.2
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    • pp.217-228
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    • 2003
  • Co-ordinate growth of the brain and skull is achieved through a series of tissue interactions between the developing brain, the growing bones of the skull and the sutures that unite the bones. Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of these interactions. Bmp2, one of bone morphogenetic proteins (Bmps), is involved in the regulation of the shapes of individual bones and the relative proportions of the skeleton. Mutations in the homeobox gene Msx2, known as a downstream gene of Bmp, cause Boston-type human craniosynostosis. The phenotype of Dlx5 homozygote mutant mouse presents craniofacial abnormalities including a delayed ossification of calvarial bone. These facts suggest important roles of Bmp2, Msx2 and Dlx5 genes in the cranial bone growth and suture morphogenesis. To elucidate the function of these molecules in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of Bmp2(E15-18), Msx2 and Dlx5 genes in the developing sagittal suture of calvaria during the embryonic stage. Bmp2 mRNA was intensely expressed in the osteogenic fronts and also at the low level in the periosteum of parietal bones during embryonic stage, Msx2 mRNA was intensely expressed in the sutural mesenchyme and mildly expressed in the dura mater during the embryonic stage. Dlx5 mRNA was intensely expressed osteogenic fronts and parietal bones. To further examine the role of Bmp signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of Bmp2-soaked beads onto the osteogenic fronts after 48 hours organ culture resulted in the increase of the tissue thickness and cell number around Bmp2 beads, compared to BSA control beads. In addition Bmp2 induced etopic expressions of Msx2 and Dlx5 genes. On the other hand, overexpression of FGF2 did not induce the expression of Msx2 and Dlx5. Taken together, these data indicate that Bmp2 signaling molecule has a important role in regulating the cranial bone growth and early morphogenesis of cranial suture. We also suggest that Bmp signaling is involved in all the stages of osteogenesis of cranial bones and the maintenance of cranial suture by regulating Msx2 and Dlx5 genes, and that Msx2 and Dlx5 genes are specific transcription factors of Bmp signaling pathway.

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Purification of cystic fluid antigen of Taenia solium metacestodes by affinity chromatography using monoclonal antibody and its antigenic characterization (단세포군항체에 의한 유구낭미충 낭액 특이항원의 순수분리 및 항원특성 관찰)

  • Kim, Suk-Il;Kang, Shin-Yong;Cho, Seung-Yull;Hwang, Eung-Soo;Cha, Chang-Yong
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.145-158
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    • 1986
  • This study was undertaken to purify cystic fluid (CF) antigen of Taenia solium metacestodes by affinity chromatogaphy using specific monoclonal antibody(McAb) and to characterize the antigenicity of the purified antigen. The hybridoma cell lines, prepared by fusion between mouse plasmacytoma and spleen cells from BALB/c mice immunized with CF, secreted antibodies reacting to various helminthic antigens. Majority of cell lines reacted to CF only but some also reacted to parenchymal antigen of T. solium metacestodes, adult T. saginata, sparganum, hydatid cystic fluid, Paragonimus westermani and Clonorchis sinensis, either in combination with CF, other antigens or independently. Cloned cells derived from monoclonal lines also produced antibodies reacting either to CF only or to other helminthes in combination or independently. These results indicated that CF of T. solium metacestodes contained proteins which possessed antigenic determinants not only specific to CF but also cross reactive with the afore-mentioned helminthes. CF of T. solium metacestodes was purified by affinity chromatography using the McAb which reacted to CF and parenchymal antigens. The affinity-purified antigen (A-Ag) and unbound pool CF (U-Ag) were separated. A-Ag showed 2 protein bands by disc-PAGE whereas CF exhibited 6 bands and U-Ag consisted of all bands CF had. The diagnostic significance of A-Ag was evaluated by ELISA in human neurocysticercosis and other helminthic and neurologic diseases. By A-Ag, the levels of the specific IgG antibody, as shown by absorbance in sera and CSF, were lower than those of CF and U-Ag. Accordingly, the sensitivity was about 70% of CF and U-Ag. However, the nonspecific positive reactions to CF and U-Ag, observed in sparganosis, T. saginata infection and paragonimiasis did not occur when A-Ag was used. These results indicated that the affinity-purified A-Ag had the higher specificity but the lower sensitivity as a diagnostic antigen in cysticercosis, probably because it only detected a single or limited numbers of monospecific antibodies among the diverse polyclonal antibodies produced in the patients with neurocysticercosis.

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Inhibitory Effect of Kailan (Brassica oleracea L.) Extract on LPS-Induced Inflammatory Response in Macrophages (카이란(Brassica oleracea L.) 추출물의 대식세포 내에서 LPS에 의한 염증 반응 억제 효과)

  • DanHee Yoo;In Chul Lee
    • Microbiology and Biotechnology Letters
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    • v.52 no.3
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    • pp.255-263
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    • 2024
  • In this study, antioxidant and anti-inflammatory activity was studied to confirm the value of kailan (Brassica oleracea L.) as a natural material for cosmetics. For this study measure the antioxidative activity, total polyphenol content was measured, and DPPH and ABTS scavenging activity assays were conducted. As a result of measuring the total polyphenol content of hot water extract of kailan (KRD) and 70% ethanol extract of kailan (KRE), it was found to contain 124.3 mg TAE/100 g and 144.1 mg TAE/100 g, respectively. As a result of DPPH and ABTS radical scavenging ability, it was confirmed that the efficacy was concentration dependent. After treating the cells with LPS, a stimulant, for 2 hours, an experiment was conducted by treating RAW 264.7 cells with KRD and KRE at concentrations of 10, 50, and 100 ㎍/ml. The nitric oxide production inhibitory activity of KRD and KRE showed an inhibitory effect of about 30% at a concentration of 100 ㎍/ml. Cells cultured for 18 hours after stimulant treatment were obtained and used in experiments. The cells obtained in this way were lysed, protein and mRNA were extracted, and the expression of inflammatory mediators' inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was confirmed. It was confirmed that the protein mRNA expression of iNOS and COX-2, measured through western blot and reverse transcription-PCR, was inhibited in a concentration-dependent manner. Based on this, it is judged that has the potential to be used as a natural material in cosmetics.