• Title/Summary/Keyword: in vivo experiment

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Action mechanism of upstream open reading frame from S-adenosylmethionine decarboxylase gene as a in vivo translational inhibitor (S-Adenosylmethionine decarboxylase 유전자의 upstream open reading frame이 in vivo에서 translational inhibitor 로서의 작용 기작)

  • Choi, Yu-Jin;Park, Ky-Young
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.87-93
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    • 2011
  • S-Adenosylmethionine decarboxylase (SAMDC; EC 4.1.4.50), a key enzyme for polyamines biosynthesis, was tightly regulated for homeostatic levels. Carnation SAMDC gene (CSDC9) has an small upstream open reading frame (uORF) of 54 amino acids in 5'-leader sequence. To explore the functional mechanism of uORFs in controlling translation, we used a GUS reporter gene driven with the 35S promoter and uORF region of SAMDC gene for making transgenic tobacco plants. In our experiment, there were a translational inhibition of its downstream GUS ORF by SAMDC uORF sequence or SAMDC uORF protein. Expecially, translational inhibition was most effective in point-mutated construct, in which the start codon was changed. Therefore, this results suggested the ribosomal stalling might be involved in this translational inhibitory process. The frame shift in amino acid sequence of SAMDC uORF with start codon and stop codon resulted in a moderate increasing in GUS activity, suggesting the native amino acid sequence was important for a function as a translational inhibitor. Also, we showed that the production of GUS protein was significantly inhibited in the presence of the small uORF using histochemical analysis of GUS expression in seedlings and tobacco flowers. Importantly, the small uORF sequence induced a real peptide of 5.7 kDa, which was provided the presence of SAMDC uORF peptide band using an in vitro transcription/translation system. The peptide product of uORF might interact with other components of translational machinery as well as polyamines, which was resulted from that polyamine treatment was inhibited GUS protein band in SDS-PAGE experiment.

The Effects of Alismatis Rhizoma Extract on Allergic Inflammation in RBL-2H3 Mast Cells and OVA/alum-Sensitized Mice (택사 (澤瀉, Alismatis Rhizoma) 추출물이 RBL-2H3 비만세포와 OVA/alum 감작 생쥐의 알레르기 염증 반응에 미치는 영향)

  • Song, Ji Hyun;Lee, Jin Hwa;Kim, Eun Jin;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.32 no.3
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    • pp.1-15
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    • 2018
  • Objectives Alismatis Rhizoma has been known to suppress inflammation and allergic reaction. However, the cellular target of Alismatis Rhizoma and its mechanism of action remain unclear. This study was designed to examine the effect of Alismatis Rhizoma extract (ALC) on the RBL-2H3 mast cells in vitro and on the OVA/alum sensitized mice ex vivo. Methods In the study, RBL-2H3 mast cells were cultured in minimal essential medium (MEM) for 24 hours, and treated separately with cyclosporin A and varying doses of ALC, and then stimulated with Phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) and Ionomycin ($0.5{\mu}M$). The levels of IL-13, IL-4 were measured by ELISA analysis. The mRNA levels of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$ were analyzed with Real-time PCR. Also, manifestations of MAPKs transcription factors and $NF-{\kappa}B$ p65 translocation were analyzed by western blotting in vitro. Subsequently, for ex vivo experiment, we induced allergic inflammation on Balb/c mice by OVA/alum and administered ALC orally. And we measured serum OVA-specific IgE level and IL-4, IL-13 in the splenocyte culture supernatant by ELISA analysis. Results ALC was shown to suppress mRNA expression of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$, and to inhibit the IL-13, IL-4 production. Also ALC reduced an activation of mast cells specific signal MAPKs transcription factors and $NF-{\kappa}B$ p65 from the western blot analysis in in vitro experiment. In ex vivo, ALC oral adminstration decreased the level of OVA-specific IgE in serum, and IL-4, IL-13 in the splenocyte culture supernatant. Conclusions ALC is shown to reduce inflammation and allergic response by suppressing Th2 cytokines through the regulation of transcription factors MAPKs and $NF-{\kappa}B$ p65 in mast cells. Administration of ALC suppressed OVA-specific IgE in ovalbumin allergy model through the inhibition of Th2 cytokine. In conclusion, ALC can be considered as an effective treatment for allergic diseases such as atopic dermatitis.

Calcium Absorption Acceleration Effect on Phosphorylated and Non-phosphorylated Peptides from Hoki (Johnius Belengeri) Frame (민태 Frame 단백질 유래의 인산화 및 비인산화 펩타이드가 체내 칼슘 흡수에 미치는 영향)

  • KIM Se-Kwon;JEON You-Jin;BYUN Hee-Guk;PARK Pyo-Jam;KIM Gyu-Hyung;CHOI Yong-Ri;LEE Yeon-Sook
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.6
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    • pp.713-717
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    • 1999
  • In order to utilize protein hydrolysate produced from hoki (Johnius Belengeri) frame among many different fish processing wastes, hoki frame peptide (PHFP) and phosphorylated hoki frame peptide (PHFP) were prepared, and their calcium absorption accelerating effects were investigated in comparison to control and casein phosphopeptide (CPP). In in vitro experiment, HFP and PHFP inhibited calcium phosphate formation as high as 1.5-fold and 2.5-fold, respectively, comparing to control, In addition, the inhibition rate of calcium phosphate precipitation as increasing concentrations of HFP and PHFP was risen and was similar to that of CPP at 2.0 mg/ml of PHFP concentration, In in vivo experiment using the rats, the groups fed HFP and PHFP indicated significantly increased calcium content in the femur. In particular, the calcium content in the small intestine of the rat fed PHFP was higher than that of control group by approximately $60\%$.

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Effect of Jeseupwiryeongtang-gagam on Atopic Dermatitis-like Skin NC/Nga Mouse Induced by Dermatophagoides Farinae Crude Extract (제습위령탕가감이 집 먼지 진드기 추출물로 유도된 NC/Nga mouse의 아토피 피부염에 미치는 영향)

  • Na, Dong-Gyu;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.22 no.2
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    • pp.81-114
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    • 2008
  • Objectives : The purpose of this study is to investigate the effect of Jeseupwiryeongtang-gagam(JWTG) on atopic dermatitis by in vivo experiment using NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the atopic dermatitis of human. Methods : To investigate the effect of JWTG on AD, we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells(PBMCs), splenocytes, draining lymph node(DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis-like skin NC/Nga mouse in vivo. Results and Conclusions : In vivo, clinical skin severity score were significantly lower in JWTG group than control group. IgE, IL-6, $TNF-{\alpha}$, IgM, IgG2a and IgG2b levels in serum were decreased remarkably in JWTG group than control group and $IFN-{\gamma}$ production, secreted in Th1 cell were increased by JWTG. After experiment ended, we analyzed immunological cells ($CD3^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3^+$$CD69^+$, $CD4^+$$CD25^+$ and $CD49b^+$) by flow cytometry. It resulted that total absolute number of $CD3^+$, $CD19^+$, $CD4^+$ and $CD8^+$ cells were recovered as normal and $CD3^+$$CD69^+$ were decreased significantly compared with control group in isolated DLN and PBMCs from NC/Nga mouse and total absolute number of $Gr-1^+$, $CD11b^+$ and $CD3^+$ in dorsal skin of NC/Nga mouse were decreased by JWTG. We analyzed ear, DLN, and neck-back skin after biopsy and dyeing by hematoxyline/eosin(H&E) and toluidine staining (mast cells marker) and obtained results that JWTG were effective to histological symptoms (dermal and epidermal thickening, hyperkeratosis and inflammatory cell infiltration). Ear thickness was decreased significantly than the control group and the size of inflammatory lymphocytes cells(ILC) and plasma cells(PC) in DLN were also decreased.

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The Effect of Squalene on Inflammation Factors Induced by Candida Albicans in Vivo Studies (생체 내 상에서 Candida albicans로 유도한 염증인자에 미치는 스쿠알렌의 효과)

  • Lee, Jun-Haeng
    • Journal of radiological science and technology
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    • v.39 no.3
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    • pp.443-449
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    • 2016
  • In the present study, whether squalene treatment relives inflammatory reactions induced by Candida albicans was checked. The experiment was conducted in vivo using seven experimental animals (ICR mice) per experimental group. Among C. albicans-induced inflammatory factors, TNF-${\alpha}$, IL-6, and NO were observed using the ELISA kits method. Through the experiment, the following conclusions were obtained. 1. In the group infected with C. albicans, it could be identified that squalene treatment was inducing NO generation in renal tissues both on the 1st and 3rd days (p < 0.05). 2. In the group pre-treated(intraperitoneal administration) with SQ (80ml/kg) once per day for seven days and infected with C. albicans, it could be identified that squalene treatment was inducing TNF-${\alpha}$ generation in renal tissues only on the 3rd day(p < 0.05). 3. In the group pre-treated(intraperitoneal administration) with SQ (80ml/kg) once per day for seven days and infected with C. albicans, it could be identified that squalene treatment was inducing IL-6 generation in renal tissues only on the 3rd day(p < 0.05). In conclusion, it could be seen that for squalene to suppress C. albicans-induced inflammatory factors, preemptively supplying SQ should be effective. Therefore, effects for recovery from C. albicans-induced immunodepression can be expected from SQ treatment.

Evaluation of Electrohydraulic Left Ventricular Assist Device through Animal Experiment (동물실험을 통한 전기유압식 좌심실 보조장치의 성능 평가)

  • Choi, J.S.;Chung, C.I.;Choi, W.W.;Park, S.K.;Jo, Y.H.;Om, K.S.;Lee, J.J.;Won, Y.S.;Kim, H.C.;Kim, W.G.;Min, B.G.
    • Proceedings of the KOSOMBE Conference
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    • v.1996 no.05
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    • pp.84-87
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    • 1996
  • We have been developed electrohydraulic left ventricular assist device and done various in vivo evaluation on the device. Through the in vivo experiment conducted from Jan. 23, 1996 to Feb. 8, we could have experience of long-term evaluation fur the first time. The sheep used in this experiment had survived for 16 days. We used new actuator with reduced size and linear motion guide replacing oil box and ball bearings. Also, we used improved blood chamber with reduced size, reduced weight facilitating fixing the chamber to animal's body, and polymer sac having improved folding pattern. Against suction problem, we used absolute pressure limiter only. Motor current for driving this new actuator was not much higher than older one. Effective stroke volume was about 48 cc. Thrombosis was found around top area and peripheral boundary of the sac and valves. There was no sign of damage from suction problem in the atrium observed at autopsy. Main cause of death was presumed to be progressive formation of thrombosis in the cannulae. In this paper, the results of this experiment are documented.

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Direction of Tissue Contraction after Microwave Ablation: A Comparative Experimental Study in Ex Vivo Bovine Liver

  • Junhyok Lee;Hyunchul Rhim;Min Woo Lee;Tae Wook Kang;Kyoung Doo Song;Jeong Kyong Lee
    • Korean Journal of Radiology
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    • v.23 no.1
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    • pp.42-51
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    • 2022
  • Objective: This study aimed to investigate the direction of tissue contraction after microwave ablation in ex vivo bovine liver models. Materials and Methods: Ablation procedures were conducted in a total of 90 sites in ex vivo bovine liver models, including the surface (n = 60) and parenchyma (n = 30), to examine the direction of contraction of the tissue in the peripheral and central regions from the microwave antenna. Three commercially available 2.45-GHz microwave systems (Emprint, Neuwave, and Surblate) were used. For surface ablation, the lengths of two overlapped square markers were measured after 2.5- and 5-minutes ablations (n = 10 ablations for each system for each ablation time). For parenchyma ablation, seven predetermined distances between the markers were measured on the cutting plane after 5- and 10-minutes ablations (n = 5 ablations for each system for each ablation time). The contraction in the radial and longitudinal directions and the sphericity index (SI) of the ablation zones were compared between the three systems using analysis of variance. Results: In the surface ablation experiment, the mean longitudinal contraction ratio and SI from a 5-minutes ablation using the Emprint, Neuwave, and Surblate systems were 28.92% and 1.04, 20.10% and 0.53, and 24.90% and 0.45, respectively (p < 0.001). A positive correlation between longitudinal contraction and SI was noted, and a similar radial contraction was observed. In the parenchyma ablation experiment, the mean longitudinal contraction ratio and SI from a 10-minutes ablation using the three pieces of equipment were 38.60% and 1.06, 32.45% and 0.61, and 28.50% and 0.50, respectively (p < 0.001). There was a significant difference in the longitudinal contraction properties, whereas there was no significant difference in the radial contraction properties. Conclusion: The degree of longitudinal contraction showed significant differences depending on the microwave ablation equipment, which may affect the SI of the ablation zone.

Development of Pediatric VAD(VICT) and its in-vivo Test (소아용 좌심실 보조장치(VICT)의 개발 및 in-vivo 실험)

  • Lee, S.H.;Park, I.T.;Kim, S.H.;Lee, K.W.;Lee, K.H.;Ahn, H.
    • Proceedings of the KOSOMBE Conference
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    • v.1996 no.05
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    • pp.207-212
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    • 1996
  • We developed pneumatic blood pump and its driving system as a pediatric ventricular assist device. The blood pump is diaphragm type system and its blood contacting area is coated with Bio-Span. The driving unit Is consists of dual pumps, valves for the reliable blood pumping and its controller uses 80C196(Intel) as a main processor. The acute animal experiment was performed with dogs and its body weight is about 20 kg. The maximum cardiac output is about 2.1 l/min and the pressure and flow curves showed reliable operation as assist device.

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Nutrient Requirements of Exercising Swamp Buffalo, Bubalus bubalis, from Materials Balance and In Vivo Body Composition by the Body Density Method. I. Aspects of Energy and Protein Metabolism in Working Cows

  • Mahardika, IG.;Sastradipradja, D.;Sutardi, T.;Sumadi, IK.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.5
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    • pp.605-612
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    • 2000
  • Four young swamp buffalo cows of similar age ranging in weight between 280 to 380 kg and trained to do physical work were used in a study to determine energy and protein requirements for draught using a $4{\times}4$ Latin square designed experiment. The experiment consisted of field trials employing 4 levels of work load, e.g. no work as control, and loads amounting 450 to 500 Newton (N) pulled continuously for 1, 2 and 3 h daily for 14 consecutive days. Cows were fed king grass (Penisetum purpuroides) ad libitum and were subjected to materials balance trials. Body composition was estimated in vivo by the body density method and daily energy expenditure (EE) was calculated from ME minus retained energy (RE). The results show that EE while not working ($EE_{resting}$) was $0.42kgW^{0.75}MJ/d$ and maintenance ME ($ME_m$) was $0.37kgW^{0.75}MJ/d$. ME requirement increased to 1.65 times maintenance for the work of 3 hours. The energy expended for doing exercise ($E_{exercise}$) was 9.56, 20.0 and 25.86 MJ/cow for treatments 1, 2 and 3 II, respectively. Fat retention was absent in all groups of working cows, but protein retention was only negative for cows undertaking 3 h work. The relationship between $E_{exercise}$ (MJ), work load (F, kN), work duration (t, h) and body mass (W, kg) was found to be: $E_{exercise}=(0.003F^{1.43}t^{0.93})/W^{0.09}MJ$. The maintenance requirement for digestible protein was $2.51kgW^{0.75}g/d$, whereas digestible protein for growth ($DP_{growth}$) and for work ($DP_{work}$) followed the equations: $DP_{growth}=[(258+1.25W^{0.75}){\Delta}Wkg/d]g$ and $DP_{work}=[12.59e^{0.95t}]g$, respectively The coefficients a, b and c for the calculation of $E_{exercise}$ components according to the Lawrence equation were found to be 2.56 J/kgW.m, 5.2 J/kg load carried.m and 0.29, respectively, thus efficiencies to convert ME into work were 0, 16.09, 27.3 and 32.44% for control, 1, 2 and 3 h/d work, respectively. ME and DP requirements for a 250 to 400 kg working buffalo cow allowing to growth up to 0.5 kg/d are presented.

Evaluation of Biocompatibility of Extracorporeal Circuit - Development of a Quantification Technique using in-vivo Injection of Tc99m Radioactive Platelets - (체외순환도관의 혈액적합성 평가 - 방사선 동위원소(Tc99m) 활성화 혈소판의 생체 내 주입을 이용한 정량분석법의 개발 -)

  • Lee, Sung-Ho;Sun, Kyung;Choi, Jai-Geol;Son, Ho-Sung;Jung, Jae-Seung;Ahn, Sang-Soo;Oh, Hye-Jung;Lee, Whan-Sung;Lee, Hye-Won;Kim, Kwang-Taik;Jeong, Yoon-Seop;Kim, Young-Ha;Kim, Hyoung-Mook
    • Journal of Chest Surgery
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    • v.35 no.3
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    • pp.171-176
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    • 2002
  • Background: Blood-foreign interaction cause activation of coagulation and inflammatory process that may lead to multiorgan dysfunction and determine the surgical outcomes. Of the methods for assessing the biocompatibility, the platelet adhesion study is considered as the most valuable evaluation step in blood-foreign interaction. As the most studies have used in-vitro or ex-vivo conditions, we have developed a technique of quantification for platelet adhesion on the blood contact surface by using in-vivo injection of radioactive platelets. Material and Method: A coupled bypass circuit was designed to connect the proximal and descending thoracic aorta in 6 piglets(20∼25 Kg). One side of the circuit tube was consisted of a heparin coated PVC tube(10mm in ID, n=6, Experimental group), and the other, a non-heparin coated PVC tube(10mm in ID, n=6, Control group). After cannulation, the blood was circulated through the circuit for 2 hours. Platelet concentrate was prepared from homologous pig blood 24 hours before the experiment. The platelet concentrate was incubated with Tc-99m-HMPAO for 30 min and then centrifuged for 10 min. The supernatant was discarded and the radio-labeling efficacy was measured. The radio-labeled platelet concentrate was mixed with the autologous plasma to make the volume 5 ml, and the mixture was injected intravenously into the experimental animal. After 2 hour circulation, 5 pieces of the specimen(10mm in length each) were obtained from each PVC tube. The radioisotopes were counted with a gamma counter(Cobra ll, Packard, USA), and the ratio of radioisotope count was compared between the control and experimental group. Result: The radioisotope count number was 537.3221.1 Ci/min in the control group and 311.1 184.5 Ci/min in the experimental group(p=0.0104). The ratio between the groups was 1 to 0.58 (p=0.004). Conclusion: In vivo quantification using technetium-99m-HMPAO labeled platelets is simple and reproducible in evaluating platelet adhesion on a foreign surface. We suggest this technique to be a useful tool for blood compatibility test.