Kim Jae Chul;Park In Kyu;Lee Kyu Bo;Sohn Sang Kyun;Kim Moo Kyu;Kim Jung Chul
Radiation Oncology Journal
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v.17
no.2
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pp.151-157
/
1999
Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.
Kee, Hyung Min;Yi, Dae Yong;Yun, Ki Wook;Lim, In Seok;Ha, Tae-Seon
Childhood Kidney Diseases
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v.18
no.1
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pp.36-41
/
2014
Purpose: Urinary tract infections (UTIs) are the most common source of bacterial infections in infants and young children. Accurate diagnosis and treatment is important because of their association with renal scarring, which can lead to complications. Urine endothelin-1 (ET-1) is the major renal isoform produced and released by renal mesangial cells in response to glomerular injury. This study aimed to investigate whether urinary levels of ET-1 can be used as a biomarker for UTI diagnosis. Method: We conducted a prospective study using medical records of 70 patients below the age of 18 years, who visited Chung-Ang University Hospital from July 2012 to July 2013. We classified the patients into the UTI and control groups based on urine culture studies. The UTI group was further divided into upper and lower UTI groups using 99m-Technetium dimercaptosuccinic acid scintigraphy. Urine ET-1 was measured using enzyme linked immunosorbent assay with 0.3 mL urine. Results: The UTI and control groups were comprised of 45 and 25 patients, respectively. Mean urine ET-1 levels were significantly higher in the UTI group than in the control group ($1.41{\pm}0.35$ pg/mL vs. $0.33{\pm}0.07$ pg/mL, P =0.04). There was no significance difference in the quantitative value between the upper and lower UTI groups (P =0.552). There was no correlation between urine ET-1 and serum C-reactive protein (Pearson correlation [R]=0.24), urine ET-1 and serum white blood cell count (R=0.19). Conclusion: Our study suggests that urine ET-1 can be used for early diagnosis of UTI in children.
There is an increasing number of reports describing Streptococcus parauberis as an important pathogen of cultured olive flounder Paralichthys olivaceus and starry flounder Platichthys stellatus in Korea. We tried to determine the effects of water temperature (14${^{\circ}C}$ and 21${^{\circ}C}$) on the pathogenicity in Streptococcal disease caused by S. parauberis. We have challenged 180 olive flounder by i.p injection to $2.0{\times}10^{7}$ live cells/fish. Mortality was monitored for 21 days post challenge. And histopathological characterizations as infection degree, tissue degeneration and/or bacterial distribution were investigated with H&E stain and in situ hybridization technique. Fifty percent and 16.7% of mortality occurred within 21 days at 21${^{\circ}C}$ and 14${^{\circ}C}$ water temperature, respectively. In most cases, the typical symptoms of olive flounder infected with S. parauberis were darkness of the skin, lethargy, mild abdominal distension cause by ascites, splenomegaly, congested liver and internal organs paleness. The pericardial sac contained large amounts of cloudy fluid. Numerous whitish nodules, which were variable in size and often confluent, were randomly scattered throughout the myocardium. Especially, pericarditis and/or myocarditis was observed in all tested fishes after death. Positive in reaction with S. parauberis were found in all tissues in situ hybridization analysis. The relative numbers of S. parauberis in heart were much more than in liver, spleen, kidney and stomach. We evaluated that S. parauberis strain causes serious damage in the pericardium, shortness of breath and the blood disorder. Therefore, pericarditis and myocarditis caused by S. parauberis were closely related to mortality of olive flounder.
Objectives : Iron (Fe) is an essential element in biological processes; however excessive Fe is harmful to human health. Some air pollutants contain a high level of Fe, and the human lung could therefore be over-exposed to Fe through inhaled air pollutants. This study was performed to investigate the role of metal transporters (divalent metal transporter 1, DMT1, and metal transporter protein 1, MTP1) in the lung under the environments of Fe deficiency in the body and Fe over-exposure in the lung. Methods : Rats were fed Fe deficient (FeD, 2-6 mg Fe/kg) or Fe supplemented (FeS, 120 mg Fe/kg) diet for 4 weeks, followed by a single intratracheal instillation of ferrous sulfate at low (10 mg/kg) or high (20 mg/kg) dose. Fe concentration was analyzed in the serum, lung and liver, and histopathological findings were observed in the lung at 24 hours after Fe administration. The level of DMT1 and MTP1 expression in the lung was analyzed by RT-PCR. Also, the effect of Fe deficiency in the body was evaluated on the level of Fe concentration and metal transporters compared to FeS-diet fed rats at the end of 4-week FeD or FeS diet. Results : The 4-week FeD diet in rats induced an Fe deficiency anemia with decreased serum total Fe, increased unsaturated Fe binding capacity and hypochromic microcytic red blood cells. The concentration of Fe in the lung and liver was lower in the FeD-diet fed rats than in the FeS-diet fed rats. The level of metal transporters mRNA expression was higher in the FeD-diet fed rats than in the FeS-diet. The concentration of Fe in the lung was increased in a dose-dependent pattern after intratracheal instillation of Fe into the rats, while the level of Fe in the serum and liver was not increased in the low-dose Fe administered rats. Therefore, DMT1 and MTP1 mRNA was highly expressed in both FeD-diet and FeS-diet fed rats, after intratracheal instillation of Fe. Conclusions : DMT1 and MTP1 mRNA were more highly expressed in FeD-diet fed rats than in FeS-diet fed rats. The over-exposure of Fe intratracheally induced high expression of metal transporters and increased Fe deposition in the lung in both FeD-diet and FeS-diet fed rats, but did not increase the Fe level of the serum and liver in low-dose Fe administered rats. These results suggest that the role of metal transporters in the lung might be different in a part from the duodenum under the environment of over-exposure to Fe.
In an attempt to investigate the effect of Hymenolepis dana infection on immunological responses to sRBC in ICR strain of mice, cellular and humoral immune responses were chronologically monitored after sensitization with sRBC. Mice weighing about 20 g were allocated into artificial and natural infection groups. The shell-free eggs of H. dana were inoculated into mice on the day 0 (initial) and day 10 in the former group, and praziquantel (25 mg/kg/day) was administered for 3 days to the one half of the mice at the 15th day after the first inoculation and to all of the mice in natural infection group. In artificial infection group, the delayed-type hypersensitivity (DTH) to sRBC was considerably decreased on the day 10 after the first inoculation, and then elevated gradually to normal. Eosinophils in the peripheral blood increased slightly. The hemagglutinin (HA) and hemolysin (HE) titers during the early stage were shown to be more or less higher than those of control. Thereafter, the titers were returned to normal, followed by a transient decrease on the day 15 post-infection. The sRBC rosette and antibody-treated rosette-forming capacities on the day 15 post.infection were temporarily lowered but became higher thereafter. The mucosal mast cells (MMC) in the small intestine were gradually increased to make a peak on the day 10 post-infection and then maintained more or less at lower level. After praziquantel treatment, the DTH and the number of eosinophils were decreased slightly and the MMC number and sRBC rosette-forming capacity were considerably decreased. The titers of HA and HE and antibody-treated rosette-forming capacity, however, were elevated in general. In natural infection group, the DTH, the number of eosinophils, and MMC which were elevated due to H. dana infection were gradually returned to normal after prasiquantel treatment. The titers of HA and HE which were decreased by parasite infection were increased to normal after the treatment. However, the capacities of sRBC rosette or antibody-treated rosette formation were maintained at low levels in spite of the treatment. These results revealed that the immune responses to sRBC were significantly activated during H. dana infection, although they were transiently decreased during the days 10~15 post-infection.
The search for tumor-avid agents for use in nuclear medicine imaging or therapy is a field of ongoing importance. Metallothionein (MT) is an intracellular protein that binds many metals with isotopes having imaging or radiotherapeutic potential. The purpose of the study was to determine whether uptake of radioisotopes that bind to MT is increased in tumor. We measured the uptake of Cd-109 and Ga-67 in tumor and normal tissues of sarcoma-bearing mice. Tumors were grown subcutaneously in female Balb/C mice from cultured Balb/3T3 cells transformed by the Moloney murine sarcoma virus (MMSV). When the tumors reached about 1 cm in diameter, mice were injected subcutaneously with Cd-109 and Ga-67. Eighteen and seventy-two hours later, the mice were sacrified. Organs and tissues were removed, weighed, and activity per mg tissue determined by gamma well-counting. Uptake of Cd-109 by MMSV tumors exceeded that by normal tissues examined, with the exception of liver and kidney (the organs known to be richest in MT). The tumor-to-tissue ratios of uptake for Cd-109 were far greater than those for Ga-67 for many normal tissues of great importance in terms of background activity (bone, intestine, fat, muscle, and blood). We concluded that metals that bind to MT may be useful for oncologic imaging or rediotherapy of cancer.
Background : Potassium channel opener (K-opener) opens ATP-sensitive K'-channel located at cell membrane and induces potassium efflux from cytosol, resulting in intracellular hyperpolarization. Newly synthesized K-opener is currently examined for pharmacologic potency by means of rubidium release test from smooth muscle strip pre-incubated with Rb-86. Since in-vivo behavior of thallium is similar to that of rubidium, we hypothesized that K-opener can alter T1-201 kinetics in vivo. Purpose : This study was prepared to investigate the effects of pinacidil (one of potent K-openers) on the T1-201 uptake and clearance in cultured myocyte, and in-vivo biodistribution in mice. Methods : Spontaneous contracting myocytes were prepared to imitate in-vivo condition from 20 hearts of 3-5 days old Sprague-Dawley rat and cultured for 3-5 days before use ($5{\times}10^5$ cells/ml). Pinacidil was dissolved in 10% DMSO solution at a final concentration of 100nM or l0uM and was co-incubated with T1-201 in HBSS buffer for 20-min to evaluate its effect on cellular T1-uptake, or challenged to cell preparation pre-incubated with T1-201 for washout study. Two, 40 or $100{\mu}g$ of pinacidil was injected intravenously into ICR mice at 10 min after $5{\mu}Ci$ T1-201 injection, and organ uptake and whole body retention rate were measured at different time points. Results : Co-incubation of pinacidil with T1-201 resulted in a decrease in T1-201 uptake into cultured myocyte by 1.6 to 2.5 times, depending on pinacidil concentration and activity of T1-201 used. Pinacidil enhanced T1-201 washout by 1.6-3.1 times from myocyte preparations pre-incubated with T1-201. Pinacidil treatment appears to be resulted in mild decreases in blood and liver activity in normal mice, in contrast, renal and cardiac uptake were mildly decreased in a dose dependent manner. Whole body retention ratios of T1-201 were lower at 24 hour after injection with $100{\mu}g$ of pinacidil than control. Conclusion : These results suggest that treatment with K-opener may affect the interpretation of T1-201 myocardial images, due to decreasing thallium accumulation and enhancing washout from myocardium.
Shin, Yun Kyung;Park, Jung Jun;Park, Mi Seon;Myeong, Jeong In;Hur, Young Baek
Korean Journal of Environmental Biology
/
v.32
no.3
/
pp.216-224
/
2014
Decrease in dissolved oxygen concentrations associated with temperature fluctuation is an important criteria to evaluate the mortality rate of the species. Based on this parameter, we investigated the survival rate, physiological response and histological change of warty sea squirt. It was found that the survival rate of the warty sea squirt species was 63.3% at $23^{\circ}C$ and 56.6% at $26^{\circ}C$ respectively. However, exposure of six days at $29^{\circ}C$ caused deaths among species, which indicated the 6day-$LT^{50}$ of the tested species to be $24.58^{\circ}C$ ($19.48{\sim}35.48^{\circ}C$). Further, after 11 day of exposure, the dissolved oxygen concentration has been found to decrease, with the survival rate of 20% at $4.0mg\;L^{-1}$ and deaths at $2.0mg\;L^{-1}$, thus 11day-$LC^{50}$ calculated to be $3.88mg\;L^{-1}$ ($3.29{\sim}4.57mg\;L^{-1}$). In addition, decrease in rate of oxygen consumption and excretion of ammonia was also noted at this critical water temperature and dissolve doxygen concentration. Moreover, there has been common histopathological changes were observed in warty sea squirt's gill pouch, digestive tract, and tunic as follows such as: proliferation of epithelial cells, condensation and necrosis, permeation of phagocyte and blood cell, loss of cilium and muscular fiber degeneration. Based on our study results, we suggest that these parameters can also be useful to evaluate the survival rate and physiological response in other species.
Nitric oxide (NO) is an important regulator of renal blood flow, glomerular hemodynamics, and tubule transport processes in the kidney. There is also evidence that NO is involved in cell cycle regulation and mitotic division. During development the nNOS expression pattern differs from that observed in adult animals. However, little is known about temporal and spatial patterns of nNOS expression in the developing kidney. The purpose of this study was to establish the time of expression and the distribution of nNOS in the developing rat kidney. Kidneys from 14-, 16-, 17-, 18-, and 20-day-old fetuses, 1-, 4-, 7-, 14-, and 21-day-old pups, and adult animals were preserved and processed for immunohistochemistry. In the adult kidney, nNOS was detected in the parietal epithelium of Bowman s capsule, macula densa, descending thin limb and inner medullary collecting duct. nNOS immunoreactivity appeared first in the distal tubule anlage at 15 days of gestation, and in all epithelial cells of developing thick ascending limbs (TAL) as well as macula densa of 17- and 18-day-old fetuses. From 20 days of gestation to 14 days after birth, nNOS was expressed in the newly formed cortical TAL, which are located in the medullary ray, whereas in mature TAL of juxtamedullary nephrons, nNOS immunolabeling gradually decreased in intensity and became restricted to the macula densa. In inner medullary collecting ducts, nNOS immunoreactivity appeared first at 7 days after birth in the papillary tip and gradually ascended to the border between outer and inner medulla. In the descending thin limb and parietal epithelium of Bowman's capsule, weak nNOS immunoreactivity was observed at 14 days after birth and labeling gradually increased to adult levels at 21 days after birth. These results suggest that differential expression of nNOS in the developing kidney is an important physiological regulator of renal function during kidney maturation.
Histopathological changes and methemoglobinemia of the eels (Anguilla japonica) reared at six culture farms in the vicinity of Y$\check{o}$su city were investigated under the conditions of $22.0{\sim}29^{\circ}C$, 4.33~7.33ppm of D.O., pH 6.1~8.2 and 0.03~10.1ppm of nitrite from December, 1989 to June, 1990. All of the eels showed no any abnormal state in action, color and blood, but the eels reared in pH 6.0 had planty of mucus and stickiness on their body surface. Methemoglobin levels were 1.03~9.86% of total erythrocyes in Dec. and 5.05~25.69% in June, and poikilocyte levels were 5.8~30.17% in Dec. and 8.67~74.45% in June. These results indicate the fact that the water management of the culture farms was worse in summer than in winter, even though not good in winter. Hypertrophy was found in the gill, hepatic and kidney tissue of the eels in all the culture farms. In March and June, Fish of all culture farms appeared the detachment of gill lamella epithelium cells, pyknosis and sinusoids of nucleus in the hepatic tissue and hemosiderosis in the kidney tissue. The trend of increase in methemoglobin levels appeared in the fish of culture farms with high nitrite concentration, which didn't appear equally in all the culture farms. However, in the culture farms of pH 6.0, methemoglobin and poikilocyte levels were increased independently of concentration of nitrite, and degeneration in the tissue of the gill, liver and kidney was also appeared.
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