LEE , JI-HYUN;LEE, DOO-HOON;SON, JEONG-HWA;PARK, JUNG-KEUG;KIM, SUNG-KOO
Journal of Microbiology and Biotechnology
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v.15
no.1
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pp.7-13
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2005
Chitosan-alginate capsules were formed by electrostatic interactions and exhibited an appropriate mechanical strength, permeability, and stability for the culture of hepatocytes. Pig hepatocytes were isolated and hepatocyte spheroids formed and immobilized in chitosan-alginate capsules. An encapsulation procedure of 3 min and spheroid formation period of 24 h were the optimum conditions for the best liver functions. Pig hepatocytes with a cell density of $6.0{\tomes}10^6$ cells/ml in the capsules were found to be most suitable for application in a bioartificial liver support system. The encapsulated pig hepatocyte spheroids exhibited stable ammonia removal and urea secretion rates in a bioreactor for 2 weeks. Accordingly, chitosan-alginate encapsulated hepatocyte spheroids in a packed-bed bioreactor would appear to have potential as a bioartificial liver.
The entry of antipyrine and urea from the peritoneal cavity of rabbit into organ tissue and blood plasma was studied. Two hundred mg of antipyrine plus 300 mg of urea in 10 ml Ringer's solution was injected into the peritoneal cavity of anesthetized rabbit. The injection was made from above of a rabbit kept tying right side down and it enabled part of the abdominal organs (liver, intestine, kidney) was immersed in the injected solution and kept high concentration gradient throughout the experimental period. The remaining part of the organs was revered only by a thin film of the test solution. Subsequently, in this part of the organs the concentration gradient of the diffusible substances during entry was presumed to decrease as time elapsed. Four pieces of the liver tissue were taken namely, the right superficial, right deep, left superficial and left deep portions. Two were taken from the small intestine, one from the portion which was immersed in. the fluid and the other from that above the fluid mass. Both kidneys were separately analyzed. As a remote organ the gastrocnemius muscle was taken from the right leg of the animal. The intervals which were the time periods elapsed after injections were 5,7,10,15 or 30 minutes. At each point 5 animals were sacrificed and the concentrations of the test substances in the tissue water were measured. The results obtained were as follows. 1. In the liver the right portion which was immersed in the fluid showed higher concentration if the test substances than the left portion and the superficial region exceeded the deep region. The concentrations diminished as the time elapsed after infusion, particulary in the case of antipyrine, suggesting circulatory removal of the substances. In urea such decreasing tendency of the concentration was not obvious, and suggested slower removal rate of it as compared with that of antipyrine. 2. In the small intestine there was no regional difference in the concentration of the test substances. Because of the intestinal motility different portions of the intestine were seemed to have bathed in the fluid of the same concentration. In general the concentrations in the intestinal wall exceeded those of the liver, suggesting a slower removal rate than in the latter. 3. In the kidney the accumulation of the endogenous urea was predominant, and the accumulating mechanism in the renal tissue went on during the period of the experiment. Therefore it revealed increasing tendencies as the time elapsed. The penetration of the test substances in this organ from the peritoneal cavity seemed to be slower than in other abdominal organs, namely liver or small intestine. Part of the test substances in the kidney were obviously brought by the blood stream. 4. Rapid exponential decay of the concentration of antipyrine and of the osmolality of the peritoneal fluid was attributed to the extensive removal through the whole dimension of the peritoneal surface, and the remote organ such as the gastrocnemius muscle attained a fairly close value to that of the abdominal organs in less than 30 minutes. The factors which related to the absorption rate were discussed. They were the concentration gradient, permeability and the regional perfusion rate.
The relative permeabilities, distribution coefficients and diffusion coefficient of some salts which are important components in blood plasma through a poly(HEMA) membrane were measured. The crosslinker which was used for preparing the membrane was tetraethylene glycol dimethacrylate(TEGDMA), the weight percentage of the latter was about 2.8. We found that the diffusion coefficients ($D_m$) of the solutes decrease exponentially with increasing molecular weight, and also that $D_m$'s decrease linearly (except urea) with cylindrical radius (a). These facts were explained by a sieve pore flow model. The relative permeability and diffusion coefficient of urea at various temperature were larger than those of other solutes such as glycine, ${\beta}$-alanine, D-glucose, saccharose and maleic acid. The result indicates that the poly(HEMA) membrane might be suitable for hemodialysis application.
The appearance rates of antipyrine and urea into cerebrospinal fluid from blood were studied in the rabbits which were in the state of hypotension and of high permeability in the capillary beds following injection of histamine. The alteration in the distribution of electrolytes among various compartments of the brain and the permeability characteristics in the blood-cerebrospinal fluid barrier were also observed. Adult male rabbits, weighing around 2 kg, were used. Twenty four rabbits were divided into 3 groups. Besides the control group, histamine treated rabbits were categorized into 2 groups. $H_1$ consisted of the rabbits showing moderate responses to histamine and ranging from 62 to 80 mmHg in their mean anterial blood pressure. The animals which belong to $H_2-group$ showed severe responses to histamine and the mean anterial blood pressures dropped to 30-50 mmHg. Animals were anesthetized with nembutal, 30mg/kg i.v. The mean arterial blood pressure was read by means of the mercury manometer connected to the femoral artery. The animals, treated with histamine, were kept in hypotensive state at least for 40 minutes before the administration of the test-substances. The test-substances, 300 mg of urea and 200 mg of antipyrine, were dissolved in 3 ml of distilled water and were injected into the ear vein of the rabbit. After 10 minutes elapsed arterial blood sample was taken from the femoral artery and cerebrospinal fluid from the cisterna magna. Brain tissues were also analysed with respect to electrolytes in order to observe the disturbances in the electrolytes balance as well as in the function of the central nervous system. The results obtained were as follow: 1. The ratio of antipyrine concentration in cerebrospinal fluid to that of arterial blood plasma, that was the distribution ratio, was close to unity, revealing a well established equilibrium between the compartments of blood and cerebrospinal fluid in 10 minutes. In other words, there was no diffusion barrier with regard to antipyrine. The ratios over unity which were frequently seen in the histamine treated animals were attributable to the early penetration of the substance into the cerebrospinal fluid. 2. The appearance rates of urea into the cerebrospinal fluid in the histamine treated rabbits were higher in comparison with those of in the control animals. The increasing tendency in the rates was particularly remarkable in the $H_2-group$, showing the enhanced penetration of urea across the boondary. 3. In the htisamine treated $H_2-group$ the concentration of potassium in the blood plasma and cerebrospinal fluid well exceeded the control values and showed 8.5 and 9.0 mEq/l in average, respectively. Simultaneous drops in the brain tissue water were noticed, suggesting the leakage of intracellular potassium. 4. There was a coincidence in the rising pattern of potassium in the blood plasma and in the cerebrospinal fluid of $H_2-group$ and at least partial removal of the blood-cerebrospinal fluid barrier with respect to potassium was suggested in these animals. 5. The concentration of sodium in the blood plasma or in the cerebrospinal fluid showed no significant changes following histamine injection. However, sodium in the brain tissue revealed slight elevation in the histamine treated groups. 6. The ratios of the concentrations of potassium to those of sodium, [K]/[Na] in the brain tissues, were 1.92 in the control 1.82 in the $H_1$ and 1.52 in the $H_2-group$, respectively. The marked drop in the $H_2-group$ might represent neural dysfunction in the extremely hypotensive rabbits.
We have studied the effect of polarity, current density, current duration, crosslinking density, swelling ratio, and permeation enhancers on the transdermal flux of ketoprofen from acrylamide hydrogel. Hydrogel was prepared by free radical crosslinking polymerization of acrylamide. Drug loading was made just before transport experiment by soaking the hydrogel in solution containing drug. In vitro flux study using hairless mouse skin was performed at $36.5^{\circ}C$ using side-by-side diffusion cell, and the drug was analysed using HPLC/UV system. The result showed that, compared to passive flux, the total amount of drug transported increased about 18 folds by the application of $0.4\;mA/cm^2$ cathodal current. Anodal delivery with same current density also increased the total amount of drug transported about 13 folds. It seemed that the increase in flux was due to the electrorepulsion and the increase in passive permeability of the skin by the current application. Flux increased as current density, the duration of current application and loading amount (swelling duration) increased. As the cross linking density of the hydrogel increased, flux clearly decreased. The effect of hydrophilic enhancers (urea, N-methyl pyrrolidone, Tween 20) and some hydrophobic enhancers (propylene glycol monolaurate and isopropyl myristate) was minimal. However, about 3 folds increase in flux was observed when 5% oleic acid was used. Overall, these results provide some useful information on the design of an optimized iontophoretic delivery system of ketoprofen.
Journal of the Korea Institute of Building Construction
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v.24
no.1
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pp.77-86
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2024
In this study, an evaluation was conducted for a total of 16 conditions to suggest an appropriate construction method for the construction joint of polyurea waterproofing membrane coating. It was analyzed that the longer the construction time difference, the higher the rate of water leaks through joints, and it was confirmed that water leaks could be prevented through primer construction. In addition, since the surface of polyurea exposed outdoors for a long period of time is deteriorated and weakened, it was analyzed that polishing the area increases surface damage and affects the formation of the interface. During maintenance construction, it would be desirable to apply a primer before construction, and it is believed that using the same urea-based material will ensure waterproofing stability.
Vascular endotherial growth factor (VEGF) is a potent mitogen that stimulates vascular permeability and angiogenesis and has a potential in therapeutic applications. An industrial production method that provides high yield as well as purity is needed. Researches for various factors of mild solubilization with combination of ubiquitin fusion protein to increase solubility were carried out as well as by changing pH and denaturant concentration. Usage of pET28-a bacteral expression vector in BL21 (DE3) host cell was capable of producing approximately 14 g/L VEGF fusion protein in 20L fermentor. A purification process consisting of four chromatography steps including refolding and digestion with UBP1 resulted in mild solublization under the conditions of 2M urea and pH 10.0 due to ubiquitin fusion tag protein that increases in solubility of target protein VEGF. High yield of refolding and dimerization could be obtained between two step Ni-affinity chromatography. Multimeric and misfolded proteins and endotoxin were removed by DEAE anion exchange chromatography. Final monomers were removed from dimers by gel filtration chromatography. Characterization analysis of purified dimeric VEGF was performed using SDS-PAGE and RP-HPLC with a purity of 97%.
Purpose : A single center cross sectional retrospective study was performed to compare the outcomes of different peritoneal dialysis(PD) modalities in Korean children. Methods : Among children dialyzed with PD between the year 2004 and 2007, 35 children had reliable data on PD adequacy after 3 to 15 months of dialysis. Subjects were grouped by their modalities; 17, 13 and 5 children were on continuous ambulatory PD(CAPD), continuous cyclic PD(CCPD) and nightly intermittent PD(NIPD), respectively. Body weight and height, number of patients taking anti-hypertensives and laboratory data including biochemical and hemoglobin levels were compared. Dialysis adequacy including weekly Kt/Vurea, creatinine clearance (Ccr) and daily water removal were also compared. Patients were sub-grouped by their peritoneal permeability characteristics. Results : The percentage of patients taking anti-hypertensives, monthly change in Z-scores of body weight and height and laboratory data did not differ among the groups. Patients on CAPD and CCPD showed similar dialysis adequacies. Weekly dialytic Ccr was significantly lower in the NIPD group compared to the others. But total Ccr was not different when residual renal function was added. Weekly dialytic Ccr by CAPD was significantly higher than that of CCPD in low and low-average transporters. Conclusion : We propose that modality can be selected flexibly according to the patients' preferences. And peritoneal permeability characteristics provide valuable information for adjusting PD prescriptions in ultrafiltration failure or in inadequate dialysis. Further study of other clinical performance measures should be performed to clarify the comparable outcomes in different PD modalities.
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