In horse, a single gene encodes both eCG and eLH $\beta$ subunits. The difference between eCG and eLH lies in the structure of their glycoresidues, which are both sialylated and sulfated in LH and sialylated in CG eCG consists of highly glycosyiated $\alpha$- and $\beta$-subunits and is an unique member of the gonadotropin family because it elicits response characteristics of both FSH and LH in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of gonadotropin structure-function relationships and the understanding of the molecular bases of the specific interactions of these hormones with their receptors. Thus, eCG is a dintinct molecule from the view points of its biological function and glycoresidue structures. The oligosaccharide at Asn 56 of the $\alpha$-subunit plays an indispensable role, whereas the carboxyl-terminal extension of the eCG $\beta$-subunit with its associated O-linked oligosaccharides is not improtant for, the in vitro LH-like activity of eCG. In contrast, both N- and O-linked oligosaccharides play important roles for FSH-like activity and increase FSH-like activity by removal of N- and O-linked oligosaccharides. Therefore, the dual LH- and FSH-like activities of eCG can be clearly separated by removal of either the N-linked oligosaccharide on the $\alpha$-subunit or CTP-associated O-linked oligosaccharides from its $\beta$-subunit. The glycoresidues seem to play crucial roles fer biological activities. The tethered-eCG was effciently secreted and showed similar LH-like activity to the dimeric eCG $\alpha$/ $\beta$ and native eCG. FSH-like activity of the tethered-eCG was also shown similarly in comparison with the native and wild type eCG $\alpha$/ $\beta$. Our data for the first time suggest that the tethered-eCG can be expressed efficiently and the produced product by the CHO-Kl cells is fully LH- and FSH-like activities in rat in vitro bioassay system. Our results also suggest that this molecular can imply particular models ot FSH-like activity not LH-like activity in the eCG. Taken together, these data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.
Background: In order to develop the acute heart failure model for the animal experiment of the pneumatic ventricular assist device, we decided to use young pig whose coronary artery distribution is almost the same as humans and also very cheap in price. The purpose of this study is to develop stable, reproducible acute ischemic heart failure model in swine using coronary artery ligation method. Material and Method: Five young pigs whose weights are the same as adult humans are under experiment. Each pig was under endotracheal intubation and connected to a mechanical ventilator. Through left lateral thoracotomy, we exposed the heart and induced ischemic heart failure by coronary artery ligation. The ligation began at the distal part of the left anterior descending coronary artery. After 5 minutes of initial ligation we reperfused the artery and then re-ligated. Before and after each ligation-reperfusion procedure we assessed the left ventricular end-diastolic pressure, arterial pressure, and cardiac index. We also measured left ventricular end-diastolic dimension, end-systolic dimension, fractional shortening, ejection fraction using intraoperative epicardial echocardiography. After appropriate heart failure was established with sequential (from distal part of LAD to proximal location) ligation-reperfusion-ligation procedure, we inserted the ventricular assist device and operated. Result: We established stable acute ischemic heart failure in 3 of 5 young pigs with this sequential ligation-reperfusion-ligation procedure, and could maintained 50% less ejection fraction before the procedure according to intraoperative epicardial echocardiography. We also observed no ventricular arrhythmia usually associated with simple coronary artery ligation in large animals and no cardiac arrest associated with ventricular arrhythmia or myocardial stunning. In pathologic specimen, we observed scattered ischemic myocardium in all around the ischemic field induced by coronary artery ligation. Conclusion: Under the concept of ischemic preconditioning, we developed safe and reproducible acute ischemic heart failure model in swine using sequential coronary artery ligation-reperfusion-ligation method.
Shin, Jin Young;Seo, Min Ae;Choi, Eun Jin;Kim, Jin Kyung;Seo, Eok Su;Lee, Jun Hwa;Chung, Hai Lee;Kim, Woo Taek
Clinical and Experimental Pediatrics
/
v.51
no.10
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pp.1102-1111
/
2008
Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and $30{\mu}g/mL$) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis.
There are around 7,500 manure tanks to treat the manures from pigs in Korea. In the tank, there are too much sediments deposited on the base and wall, which causes low efficiency of stock capacity and manure fermentation. In order to minimize sediments and to ferment manure effectively, we developed a 2-fluid jet mixer for mixing sediments in liquid livestock manure tank. For developing the prototype, we tested a factorial experimental system with various nozzles, and simulated CFD models with two kinds of nozzle arrangement. From the results of factorial experiment and CFD simulation, we concluded the dia. ratio of primary : secondary nozzle should be 1:2 and the nozzles should be arranged at the same distances toward to the circumferential direction. With this results, we manufactured a 2-fluid jet mixer which is consists of four 2-phase nozzles, centrifugal slurry pump and root's type air blower. And, we carried out the performance test of the prototype in the round shaped liquid manure tank in the farm. The performance test results showed that the uniformity of TS (Total Solid) and VS (Volatile Solid) was raised from 21.3 g/L, 13.3 g/L In steady state to TS and VS to 23.0 g/L, 14.1 g/L in the mixing operation. Therefore, we could conclude that the prototype of 2-fluid mixer could make the solid material which could be sediments in the tank not to be deposited in the tank and to be contacted to air bubbles which could enhance the efficiency of the fermentation of livestock manure.
A most appropriate model of 3-D conformal radiotherapy has been induced by clinical evaluation and animal study, and therapeutic gains were evaluated by numerical equation of tumor control probability(TCP) and normal tissue complication probability (NTCP). The radiation dose to the tumor and the adjacent normal organs was accurately evaluated and compared using the dose volume histogram(DVH). The TCP and NTCP was derived from the distribution of given dosage and irradiated volume, and these numbers were used as the biological index for the assessment of the treatment effects. Ten patients with liver disease have been evaluated and 3 dogs were sacrificed for this study. Based on the 3-D images of the tumor and adjacent organs, the optimum radiation dose and the projection direction which could maximize the radiation effect while minimizing the effects to the adjacent organs could be decided. 3). The most effective collimation for the normal adjacent organs was made through the beams eye view with the use of multileaf collimator. When the dose was increased from 50Gy to 70Gy, the TCP for the conventional 2-port radiation and the 5-port multidimensional therapy was 0.982 and 0.995 respectively, while the NTCP was 0.725 and 0.142 respectively, suggesting that the 3-D conformal radiotherapy might be the appropriate therapy to apply sufficient radiation dose to the tumor while minimizing the damages to the normal areas of the liver. Positive correlation was observed between the NTCP and the actual complication of the normal liver in the animal study. The present study suggest that the use of 3-D conformal radiotherapy and the application of the mathematical models of TCP and NTCP may provide the improvements in the treatment of hepatoma with enhanced results.
Journal of the Korean Society of Food Science and Nutrition
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v.41
no.8
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pp.1086-1093
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2012
The hypoglycemic and hypolipidemic effects of autoclaved soy flour and DJI chungkukjang powder fermented using Bacillus subtilis DJI were investigated in type 2 diabetic animal models. After a 2-week adaptation period, the diabetic animal model db/db mice were divided into the diabetic control group (D-C group), a diabetic group fed with soybean (D-S group), and a diabetic group fed with DJI chungkukjang (D-CJ group). The body weight gain, food intake, water intake, liver, and adipose tissue weights were not significantly different between the experimental groups. The supplementation of DJI chungkukjang or autoclaved soy flour diet induced a marked reduction of fasting blood glucose, blood glycosylated hemoglobin levels, and glucose levels in the oral glucose tolerance test and AUC for glucose compared with the diabetic control group. However, DJI chungkukjang showed a much stronger antidiabetic effect than unfermented autoclaved soy flour. Serum insulin levels were the same among the groups. The supplementation of DJI chungkukjang or autoclaved soy flour diet also significantly lowered the serum triglyceride, total cholesterol, and LDL-cholesterol levels compared with the control diabetic group, while it elevated the HDL-cholesterol level in the serum. This data suggests that the dietary supplementation of autoclaved soy flour or DJI chungkukjang may be useful in the control of blood glucose in animals with type 2 diabetes.
Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.
Based on the results that the extract of Korean mistletoe (KM-110) has immunological and anti-tumor activities and its main component is lectin called KML-U, this study was carried out to investigate the immunostimulatory and anti-tumor activities of FKM-110, fermented KM-110 with lactobacillus, as a basic study for the development of functional food with anti-tumor activity. The amount of lectin after fermentation determined by ELISA was varied with the fermentation time and kinds of lactobacillus. Cytotoxic effects of FKM-110 on the various tumor cells was significant and dependent on the concentration of KML-U and the kinds of lactobacillus. FKM-110 stimulated macrophage and resulted in the secretion of some cytokines such as IL-1 and $IFN-{\gamma}$, but this effect was not correlated with the concentration of lectin. FKM-110 fermented with Marshall Lactobacillus casei showed the most potent antitumor activity in experimental and spontaneous metastasis models. When yoghurt produced with KM-110, Marshall Lactobacillus casei and skim milk was administered orally to mouse, the metastasis of tumor cells was significantly inhibited.
Park, Sookuk;Sin, Jihwan;Jo, Sangman;Hyun, Cheolji;Kang, Hoon
Journal of the Korean Institute of Landscape Architecture
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v.44
no.4
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pp.100-108
/
2016
The climatic index for tourism(CIT) has recently been advanced, which includes complete human energy balance models such as physiological equivalent temperature(PET) and universal thermal climate index(UTCI). This study investigated human thermal sensation and comfort at Woljung-ri Beach, Jeju, Republic of Korea, in spring and summer 2015 for landscape planning and design in beach areas. Microclimatic data measurements and human thermal sensation/comfort surveys from ISO 10551 were conducted together. There were 869 adults that participated. As a result, perceptual and thermal preference that consider only physiological aspects had high coefficients of determination($r^2$) with PET in linear regression analyses: 92.8% and 87.6%, respectively. However, affective evaluation, personal acceptability and personal tolerance, which consider both physiological and psychological aspects, had low $r^2s$: 60.0%, 21.1% and 46.4%, respectively. However, the correlations between them and PET were all significant at the 0.01 level. The neutral PET range in perceptual for human thermal sensation was $25{\sim}27^{\circ}C$, but a PET range less or equal to 20% dissatisfaction, which was recommended by ASHRAE Standard 55, could not be achieved in perceptual. Only PET ranges in affective evaluation and personal tolerance affected by both aspects were qualified for the recommendation as $21{\sim}32^{\circ}C$ and $17{\sim}37^{\circ}C$, respectively. Therefore, the PET range of $21{\sim}32^{\circ}C$ is recommended to be used for the human thermal comfort zone of beach areas in landscape planning and design as well as tourism and recreational planning. PET heat stress level ranges on the beach were $2{\sim}5^{\circ}C$ higher than those in inland urban areas of the Republic of Korea. Also, they were similar to high results of tropical areas such as Taiwan and Nigeria, and higher than those of western and middle Europe and Tel Aviv, Israel.
Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4+ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.
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