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 consist 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-t63I 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 agonist-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.
Journal of Korean Society of Environmental Engineers
/
v.29
no.6
/
pp.654-661
/
2007
The goal of this research was to identify the impact of silicate polymerization on the formation of insoluble aluminiumsilicate salts which could be a cause of irreversible fouling in the membrane process by lab-scale test. For this, the amount and characteristics of precipitates that were formed in six samples with different Al and Si concentration were analyzed. And the particles was also observed by SEM-EDS(Scanning Electron Microscope - Electron Dispersion Spectrophotometer) to compare morphology and ratio of Al and Si in each precipitates. Finally the reactive and nonreactive silicate contents in the solution and precipitates were analyzed to calculate silicate form content in each fraction. The amount of precipitates was in proportion to the total concentration of both element in solution. And the amount of insoluble particle that was not dissolved in the acid solution was recorded the highest in the sample 2 of which Si concentration was lower than the saturation concentration, 50 mg/L. The content of reactive silicate in precipitates was also recorded the highest value in sample 2 of which almost silicate form was reactive. When the silicate concentration is same, that value was recorded the highest in the sample with highest Al concentration. The SEM morphology of the precipitates was similar to that of Aluminiumhydroxide and the insoluble precipitates was not dissolved in acidic solution with pH 2.7 was able to observed only in sample 2. The ratio of Al and Si in the precipitates was ranged $0.48\sim3.14$, thai of sample 2 was recorded the highest value, 3.14. It is concluded that the insoluble aluminiumsilicate could be easily formed in the solution of which silicate exist as a reactive form and coexisting Al is sufficient.
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.
Park, Chang-Kil;Hur, Gang-Min;Seok, Jung-Ho;Lee, Jae-Heun
The Korean Journal of Pharmacology
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v.27
no.1
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pp.33-43
/
1991
To study the age dependent change of Na, K-ATPase in the erythrocyte of hypertensive rat, 1-kidey 1-clip hypertensive rat was made by the removal of right kidney and partial ligation of left renal artery. After 4 weeks, aged erythrocyte fraction was separated by density gradient centrifugation, and Na, K-ATPase activity and $^3H-ouabain$ binding with ghost cell membrane and ouabain sensitive Rb-uptake with whole cell were measured. 1) In the hypertensive rats, blood pressure was significantly increased to 165.5/119.0 mmHg (systolic/diastolic). Mean corpuscular volume and membrane protein(mg) per $10^9RBC$ were decreased and hemoglobin content was increased in the aged erythrocyte. 2) Na, K-ATPase activity in the solution containing 110 mM NaCl and 10 mM KCI, was decreased in hypertensive rat, and decreased in aged erythrocyte of both group. 3) Ouabain sensitive Rb-uptake by low RbCl concentration(4 mM) was slightly decreased in aged erythrocyte compared to that in young erythrocyte of each group, but slightly increased in young erythrocyte in hypertensive rat compared to that in normotensive rat. 4) Ouabain sensitive Rb-uptake by high RbCl concentration(16 mM) was decreased about 30% to 50 % in aged erythrocyte in both group. And in hypertensive rat, especially in young erythrocyte it was significantly decreased compared to that in normotensive rats. 5) $^3H-ouabain$ binding at 0.13 or $1{\times}10^-6M$ ouabain concentration was slightly decreased in aged erythrocyte of normotensive rat, and significantly decreased in aged erythrocyte of hypertensive rats. 6) $^3H-ouabain$ binding at 6 or $64{\times}10^-6M$ ouabain concentration is slightly decreased in aged erythrocyte of both group, but significantly decreased in young and aged erythrocyte of hypertensive rats compared to that of normotensive rats. The present results suggest that (1) in the young erythrocyte of hypertensive rat, the alterations of Na-pump activity that slightly increased in weak stimulation and inhibited in strong stimulation, may be related to increased molecular activity and the decrease in the number of low affinity site without change in high affinity site, (2) in the aged erythrocyte of normotensive rat, inhibited Na-pump may be related to the change in molecular activity of pump. (3) And in the aged erythrocyte of hypertensive rat, it may be related to the decrease in the number of high and low affinity site as well as the change in molecular activity
Purpose : We describe the changes of rat glomerular epithelial cells when exposed to high levels of glucose and advanced glycosylation endproducts(AGE) in the in vitro diabetic condition. We expect morphological alteration of glomerular epithelial cells and permeability changes experimentally and we may correlate the results with a mechanism of proteinuria in DM. Methods : We made 0.2 M glucose-6-phsphate solution mixed with PBS(pH 7.4) containing 50 mg/mL BSA and pretense inhibitor for preparation of AGE. As control, we used BSA. We manufactured and symbolized five culture dishes as follows; B5 - normal glucose(5 mM) + BSA, B30 - high glucose(30 mM) + BSA, A5 - normal glucose(5 mM) + AGE, A30 - high glucose(30 mM) + AGE, A/B 25 - normal glucose(5 mM) + 25 mM of mannitol(osmotic control). After the incubation period of both two days and seven days, we measured the amount of heparan sulfate proteoglycan(HSPG) in each dish by ELISA and compared them with the B5 dish at 2nd and 7th incubation days. We observed the morphological changes of epithelial cells in each culture dish using scanning electron microscopy(SEM). We tried the permeability assay of glomerular epithelial cells using cellulose semi-permeable membrane measuring the amount of filtered BSA through the apical chamber for 2 hours by sandwich ELISA. Results : On the 2nd incubation day, there was no significant difference in the amount of HSPG between the 5 culture dishes. But on the 7th incubation day, the amount of HSPG increased by 10% compared with the B5 dish on the 2nd day except the A30 dish(P<0.05). Compared with the B5 dish on the 7th day the amount of HSPG in A30 and B30 dish decreased to 77.8% and 95.3% of baseline, respectively(P>0.05). In the osmotic control group (A/B 25) no significant correlation was observed. On the SEM, we could see the separated intercellular junction and fused microvilli of glomerular epithelial cells in the culture dishes where AGE was added. The permeability of BSA increased by 19% only in the A30 dish on the 7th day compared with B5 dish on the 7th day in the permeability assay(P<0.05). Conclusion: We observed not only the role of a high level of glucose and AGE in decreasing the production of HSPG of glomerular epithelial cells in vitro, but also their additive effect. However, the role of AGE is greater than that of glucose. These results seems to correlate with the defects in charge selective barrier. Morphological changes of the disruption of intercellular junction and fused microvilli of glomerular epithelial cells seem to correlate with the defects in size-selective barrier. Therefore, we can explain the increased permeability of glomerular epithelial units in the in vitro diabetic condition.
Split-flow thin cell fractionation (SPLITT fractionation, SF) is a particle separation technique that allows continuous (and thus a preparative scale) separation into two subpopulations based on the particle size or the density. In SF, there are two basic performance parameters. One is the throughput (TP), which was defined as the amount of sample that can be processed in a unit time period. Another is the fractionation efficiency (FE), which was defined as the number % of particles that have the size predicted by theory. Full-feed depletion mode (FFD-SF) have only one inlet for the sample feed, and the channel is equipped with a flow stream splitter only at the outlet in SF mode. In conventional FFD-mode, it was difficult to extend channel due to splitter in channel. So, we use large scale splitter-less FFD-SF to increase TP from increase channel scale. In this study, a FFD-SF channel was developed for a large-scale fractionation, which has no flow stream splitters (‘splitter less’), and then was tested for optimum TP and FE by varying the sample concentration and the flow rates at the inlet and outlet of the channel. Polyurethane (PU) latex beads having two different size distribution (about 3~7 µm, and about 2~30 µm) were used for the test. The sample concentration was varied from 0.2 to 0.8% (wt/vol). The channel flow rate was varied from 70, 100, 120 and 160 mL/min. The fractionated particles were monitored by optical microscopy (OM). The sample recovery was determined by collecting the particles on a 0.1 µm membrane filter. Accumulation of relatively large micron sized particles in channel could be prevented by feeding carrier liquid. It was found that, in order to achieve effective TP, the concentration of sample should be at higher than 0.4%.
The lipids of milk provide energy and many essential nutrients for the newborn animal. They also have distinctive physical properties that affect the processing of dairy products. Milk fat globules mainly consist of neutral lipids like triacylglycerols, whereas the globule membranes contain the complex lipids mostly, Phospholipids are a small but important fraction of the milk lipids and are found mainly in the milk fat globule membrane and other membranous material in the skim-milk phase. The milk fats of ruminant animals are characterized by the presence of relatively high concentrations of short-chain fatty acids, especially butyric and hexanoic acids, which are rarely found in milks of non-ruminants. The fatty acids of milk lipids arise from de novo synthesis in the mammary gland and uptake from the circulating blood. The fatty acid compositions of milks are usually complex and distinctive, depending on the nature of the fatty acids synthesized de novo in the mammary gland and those received from the diet in each species. The content and composition of milks from different species vary widely; presumably, these are evolutionary adaptations to differing environments. The actual process by which these globules are formed is unkonwn, but there are indications that triglyceride-containing vesicles which bleb from endoplasmic reticulum may serve as nucleation sites for globules. Recent studies on milk have centred on the manipulation of milk lipids to increase specific fatty acids, i.e. 20-carbon omega-3 fatty acids (eicosapentaenoic acid 20:5n3, decosahexaenoic acid 22:6n3) from marine sources because the fatty acids are closely associated with a decreased risk of coronary heart disease.
From August 1986 until June 1995, single aortic valve replacement was performed in 65 patients at the Chonnam National University Hospital. worthy-eight were male and 17 were female patients, ranging from 19 to 68 years of age(median : 43 years). The causes of the valve lesions were rheumatic in 29 patients (44.6%), bicuspid aortic valve in 6 patients (6.2%), endocarditis in 6 patients(6.2%), unknown in others. Concomitant surgical procedures were performed in 10 patients : repair of congenital defect in 5, pericardiectomy in 1, coronary artery bypass grafting in 1, noncoronary sinus plication in 1, Valsalva sinus aneurysmectomy in 1, subaortic membrane resection in 1 Used valves were 51. Jude-Medical valve in 42, Duromedics valve in 22, Bjork-Shiley valve in 2, Carpentier-Edward valve in 1. There were 3 hospital deaths (4.6%), and 2 late deaths (3.2%). Follow-up was 95.2% complete. The 10-year acturlal survival rate was 85.3%. Postoperative complications were low cardiac utput in 8, arrythmia in 5, valve related hemolysis in 1, cerebral infarction in 1, and gastrointestinal bleeding in 2. Reoperation was performed in 4 for surgical bleeding, in 3 for paravalvular leak. The mean improvement in New York Heart Association functional class is from 2.79 $\pm$ 0.66 preoperatively to 1.25 $\pm$ 0.49 postoperatively(p < 0.001) The change of cardiothoracic ratio from preoperative to postoperative is 0.57 $\pm$ 0.06 to 0.54 $\pm$ 0.05 (p < 0.05). The left ventricular ejection fraction change is not significant perioperatively. There are no mechanical failures. This early and intermediate-term follow-up suggests that in adults in whom valve repair is not possible, the mechanical valve is a reliable and durable prosthesis with good hemodynamic function and a low rate of thromboembolic event.
Kim, Hyun Young;Seo, Woo Duck;Seo, Kyung Hye;Lee, Mi-Ja;Choi, Sik-Won;Lee, Kwang-Sik;Kim, Sun Lim;Kang, Hyeon Jung
KOREAN JOURNAL OF CROP SCIENCE
/
v.61
no.3
/
pp.184-190
/
2016
We investigated the antioxidative and protective effects of corn silk (Zea mays L.) ethanol extracts on human HaCaT cells and erythrocytes. The NICS-2 fraction, extracted from corn silk, exhibited favorable 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activities with $IC_{50}$valuesof$13.3{\pm}0.3{\mu}g/mL$ and $14.2{\pm}0.1{\mu}g/mL$ when compared with those of ${\alpha}$-tocopherol, a positive control, with $IC_{50}=10.4{\pm}02.2$ and $22.2{\pm}3.6{\mu}g/mL$, respectively. In addition, we investigated skin protection effects of NICS extracts of corn silk in HaCaT keratinocytes. To investigate the pharmacological potential of NICS-1 and NICS-2 extracts of corn silk on UV-B-induced damage in HaCaT cells, we measured the activity of interleukin (IL) 1a. Our results showed that all the corn silk extracts inhibited the UV-B-induced activity of IL-1a. In particular, NICS-1 extracts of corn silk significantly suppressed IL-1a activity in a dose-dependent manner without inducing cytotoxicity. These results indicate that the ethanol extracts of corn silk (Zea mays L.) could function as natural cytoprotective agents and antioxidants in biological systems, particularly the skin exposed to UV radiation, by protecting cellular membrane against reactive oxygen species (ROS).
Soo Yong Lee;Seok Hyun Kim;Min Ho Ju;Mi Hee Lim;Chee-hoon Lee;Hyung Gon Je;Ji Hoon Lim;Ga Yun Kim;Ji Soo Oh;Jin Hee Choi;Min Ku Chon;Sang Hyun Lee;Ki Won Hwang;Jeong Su Kim;Yong Hyun Park;June Hong Kim;Kook Jin Chun
Korean Circulation Journal
/
v.53
no.4
/
pp.254-267
/
2023
Background and Objectives: Although the shortage of donor is a common problem worldwide, a significant portion of unutilized hearts are classified as marginal donor (MD) hearts. However, research on the correlation between the MD and the prognosis of heart transplantation (HTx) is lacking. This study was conducted to investigate the clinical impact of MD in HTx. Methods: Consecutive 73 HTxs during 2014 and 2021 in a tertiary hospital were analyzed. MD was defined as follows; a donor age >55 years, left ventricular ejection fraction <50%, cold ischemic time >240 minutes, or significant cardiac structural problems. Preoperative characteristics and postoperative hemodynamic data, primary graft dysfunction (PGD), and the survival rate were analyzed. Risk stratification by Index for Mortality Prediction after Cardiac Transplantation (IMPACT) score was performed to examine the outcomes according to the recipient state. Each group was sub-divided into 2 risk groups according to the IMPACT score (low <10 vs. high ≥10). Results: A total of 32 (43.8%) patients received an organ from MDs. Extracorporeal membrane oxygenation was more frequent in the non-MD group (34.4% vs. 70.7, p=0.007) There was no significant difference in PGD, 30-day mortality and long-term survival between groups. In the subgroup analysis, early outcomes did not differ between low- and high-risk groups. However, the long-term survival was better in the low-risk group (p=0.01). Conclusions: The outcomes of MD group were not significantly different from non-MD group. Particularly, in low-risk recipient, the MD group showed excellent early and long-term outcomes. These results suggest the usability of selected MD hearts without increasing adverse events.
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