Purpose: Uptakes of Tc-99m MIBI (MIBI) and Tc-99m tetrofosmin (tetrofosmin) in human non-small cell lung cancer A549, multidrug-resistance associated protein (MRP) expressing cell, were investigated in vitro and in vivo. Materials and Methods: Western blot analysis and immunohistochemistry were used for detection of MRP in A549 cells with anti-MRPr1 antibody. Cellular uptakes of two tracers were evaluated at $100{\mu}M$ of verapamil (Vrp), $50{\mu}M$ of cyclosporin A (CsA) and $25{\mu}M$ of butoxysulfoximide (BSO) after incubation with MIBI and tetrofosmin for 30 and 50 min at $37^{\circ}C$, using single cell suspensions at $1{\times}10^6cells/ml$. Radioactivities in supernatants and pellets were measured with gamma well counter. A549 cells were inoculated in each flanks of 24 nude mice. Group 1 (Gr1) and Gr3 mice were treated with only MIBI or tetrofosmin, and Gr2 and Gr4 mice were treated with 70mg/kg of CsA i.p. for 1 hour before injection of 370KBq of MIBI or tetrofosmin. Mice were sacrificed at 10, 60 and 240 min. Radioactivities of organs and tumors were expressed as percentage injected dose per gram of tissue (%ID/gm). Results: Western blot analysis of the A549 cells detected expression of MRPr1 (190 kDa) and immunohistochemical staining of tumor tissue for MRPr1 revealed brownish staining in cell membrane but not P-gp. Upon incubating A549 cells for 60 min with MIBI and tetrofosmin, cellular uptake of MIBI was higher than that of tetrofosmin. Coincubation with modulators resulted in an increase in cellular uptakes of MIBI and tetrofosmin. Percentage increase of MIBI was higher than that of tetrofosmin with Vrp by 623% and 427%, CsA by 753% and 629% and BSO by 219% and 140%, respectively. There was no significant difference in tumoral uptakes of MIBI and tetrofosmin between Gr1 and Gr3. Percentage increases in MIBI (114% at 10 min, 257% at 60 min, 396% at 240 min) and tetrofosmin uptake (110% at 10 min, 205% at 60 min, 410% at 240 min) were progressively higher by the time up to 240 min with CsA. Conclusion: These results indicate that MIBI and tetrofosmin are suitable tracers for imaging MRP-mediated drug resistance in A549 tumors. MIBI may be a better tracer than tetrofosmin for evaluating MRP reversal effect of modulators.
This study was conducted to determine adequate forage crop choice and optimal level of livestock manure, when different types and levels of the livestock manure were applied in corns or sorghum$\times$sorghum hybrids for the production of organic roughages by utilizing livestock manure. For the corn, yields of annual dry matter (DM) and total digestible nutrients (TDN) were highest in N+P+K-applied treatments, showing 17.3 and 11.7 ton/ha, respectively. Treatments applied 100% composted cattle manure (8.9 and 6.1 ton/ha) and 100% cattle slurry (9.4 and 7.5 ton/ ha) in contrast with chemical fertilizer-N had higher yields of DM and TDN than no fertilizer (4.8 and 2.7 ton/ha) and P+K-applied treatments (8.8 and 6.0 ton/ha). Particularly, treatments applied 150% composted cattle manure and 150% cattle slurry were markedly higher, which represented 11.4 and 7.6 ton/ha and 10.3 and 7.3 ton/ha, respectively. Crude protein (CP) contents for corns applied livestock manure ranged from 5.6 to 6.6%, which were significantly (p<0.05) higher than those of no fertilizer (3.9%) and P+K-applied treatments (5.5%). ADF (42.4%) and NDF (58.3%) contents for no fertilizer treatment were significantly (p<0.05) higher than those of other treatments. However, TDN contents were higher for livestock manure treatments than for no and/or chemical fertilizer treatments. In particular, TDN contents of treatments applied 150% composted cattle manure and 150% cattle slurry showed 72.3 and 70.8%, respectively and both treatments were significantly (p<0.05) higher than all of the other treatments. For the sorghum$\times$sorghum hybrid, yields of annual DM and TDN for 100% (12.4 and 7.4 ton/ha) and 150% (13.1 and 7.6 ton/ha) cattle slurry-applied treatments, and N+P+K-applied treatments (12.6 and 7.7 ton/ha) were significantly (p<0.05) higher than those of the others. In the others, 150% composted cattle manure (9.3 and 5.2 ton/ha) had higher annual DM and TDN yields than P+K-applied (8.4 and 4.8 ton/ha) and 100% composted cattle manure treatments (7.4 and 4.2 ton/ha), with no significant difference. Crude protein contents for sorghum$\times$sorghum hybrid applied P+K and cattle slurry were 8.8 and 8.6%, respectively. CP contents for both treatments were significantly higher than those of composted manure ($7.5{\sim}8.3%$) and no fertilizer (4.0%) treatments, but 100% livestock manure treatments had higher CP contents than 150%-applied treatments. ADF and NDF contents for N+P+K and cattle slurry-applied treatments were significantly (p<0.05) lower than the others. However, TDN contents were highest in N+P+K and cattle slurry-applied treatments, showing 61.2 and 58.3 to 59.4%, respectively. These results indicated that application of livestock manure instead of chemical fertilizer to the soil of forage crops might not only improve yields of DM and TDN, but also reduce environmental pollution by producing organic roughages through recycling of livestock manure.
An in vitro test and a broiler feeding trial were conducted to test the effect of hydrothermal treatment of wheat bran on phytate-P degradation and it’s feeding effect on performance of broilers. Hydrothermal treatment of wheat bran was carried out at 55$^{\circ}C$ with pH 5.5 buffer solution. Phytate-P content of wheat bran decreased quadrically as the wheat bran: buffer solution ratio increased from 1:0.5 to 1:5. Phytate-P degradation was not significantly affected by incubation times above 10 min., drying temperature (55$^{\circ}C$, 65$^{\circ}C$ and 75$^{\circ}C$) or pH of the buffer solution (5.5 and 7.0). A feeding trial was conducted with 240 sex separated d-old broiler chickens (Ross$^{\circledR}$). Broilers were randomly housed to 24 cages of 10 birds each. Six cages (3 of each sex) were assigned to 4 treatments: Control-normal level of non-phytate-P (NPP); LP-low NPP treatment which had 0.1% lower NPP than Control; LPWB-LP with wheat bran which provided 475 IU of plant phytase per kg diet; LPHWB-LP with hydrothermally treated wheat bran. Results of the feeding trial showed that broilers in the LP treatment gained significantly less than other treatments in starter period (1-21d) but only male broilers for growing LP gained significantly less than Control in the grower (22-35d) and overall period. There were no significant differences in weight gain among the birds of LPWB, LPHWB and Control. Feed intake during the overall period was not significantly different between LPWB and Control but that of LP was lower than LPHWB and that of LPHWB was lower than Control. Feed/gain ratio was significantly lower in LPHWB and LP than in Control and LPWP. Mortality was highest in LPHWB. Availability of crude fat, crude ash and Ca was significantly lower in LP than other treatments. Availability of P and Zn was higher in LPWB and LPHWB than in Control and LP. Availability of P, Mg and Zn was highest in LPHWP treatment. Excretion of P was significantly lower in low NPP treatments than in Control. Serum Ca level was highest whereas serum P level was lowest in LP. Tibial crude ash content was higher in wheat bran treatments, but lower in LP than Control. However, tibial Ca content was higher in Control and LP than wheat bran treatments. Tibial P content of LP and LPWB was lower than Control. However, tibial content of Fe was highest in LP. It was concluded that wheat bran, a source of plant phytase, could be used in low NPP broiler diets to prevent the depression of performance. Reduction of P excretion can be achieved concomitantly. Hydrothermal treatment of wheat bran was effective in improving utilizability of some minerals but was not effective in improving performance of broilers.
Purpose : The aim of this study was to evaluate the feasibility of 3-[$^{131}I$]iodo-O-methyl-L-${\alpha}$-methyltyrosine ([$^{131}I$]OMIMT) as an agent for tumor image. Materials and Methods : After synthesis of 4-O-methyl-L-${\alpha}$-methyltyrosine (OMAMT), OMAMT was labeled with [$^{131}I$] using Iodogen method. In vitro cellular uptake study was performed using 9 L gliosarcoma cells at various time points upto 4 hr. The biodistribution (five rats implanted with the 9 L gliosar-coma cells per group) was evaluated at 30 min, 2 hr, 24 hr after iv injection of 3.7 MBq [$^{131}I$]OMIMT or L-3-[$^{131}I$]iodo-${\alpha}$-methyltyrosine [$^{131}I$]IMT). Gamma camera images were obtained at 30 min, 2 hr and 24 hr. Results: [$^{131}I$]OMIMT uptake was 3.3 times and 2.5 times higher than [$^{131}I$]IMT uptake at 30 min and 60 min, respectively and same after 2 hr in in vitro study using 9L gliosarcoma cells. Maximum accumulation in tumor occurred at 30 min for both [$^{131}I$]OMIMT and [$^{131}I$]IMT in tumor bearing rats. The tumor uptake of [$^{131}I$]OMIMT was significantly higher than that of [$^{131}I$]IMT at early time point studied ($3.74{\pm}0.48$ vs $0.38{\pm}0.17%$ ID/g at 30 min and $2.40{\pm}0.17$ vs $0.24{\pm}0.03%$ ID/g at 2 hr, respectively, p<0.01). However, the tumor uptake of both radiolabels were not significantly different at 24 hr ($0.04{\pm}0.01$ vs $0.05{\pm}0.01%$ ID/g). Tumor was visualized as early as at 30 min in gamma camera images. Conclusion: These data suggested that [$^{131}I$]OMIMT might be a useful tumor imaging agent and has more advantage for the tumor imaging compared to [$^{131}I$]IMT
Seo, Han-Kyung;Kim, Min-Woo;Lim, Seok-Tae;Sohn, Myung-Hee
The Korean Journal of Nuclear Medicine
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v.38
no.4
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pp.300-305
/
2004
Purpose: We underwent this study to evaluate the factors which influence labeling efficiency when modified in vivo erythrocyte labeling technique was used. Materials and methods: Thirty healthy volunteers (M:F=19:11, age:$25{\pm}2$ yrs) were enrolled in this study. Totally, two hundred ten samples were obtained from them. The 1 mg of stannous pyrophosphate was injected intravenously at the beginning of labeling. After suitable tinning time (5 min, 20 min, 35 min) passed by, blood (5 mL, 3 mL or 1 mL) was withdrawn into 10 mL syringe previously containing Tc-99m (740 MBq) and anticoagulant (heparin, ACD or CPDA) through 19-gauged scalp needle. The generator ingrowth time of Tc-99m was within 24 hrs in each case. The blood samples were placed on rotating invertor during incubation (10 min, 25 min, 40 min) but some of them were not. Immediately after the conclusion of incubation, the labeled blood specimens to analyze were centrifuged. and then %Unbound Tc-99m was calculated. Statical analysis was used paired T-test and one way ANOVA with SPSS 10.0. Results: The binding efficiency at 1 mL of blood volume was $73{\pm}32%,\;91{\pm}10%$ at 3 mL and $96{\pm}7%$ at 5 mL (p<0.01). The binding efficiency at 5 min of tinning time was $45{\pm}23%,\;98{\pm}6%$, at 20 min and $97{\pm}8%$ at 35 min (p<0.001). The binding efficiency at 10 min of incubation time was $96{\pm}7%,\;95{\pm}12%$ at 25 min and $98{\pm}3%$ at 40 min (p>0.05). The binding efficiency in case of using rotating invertor was $96{\pm}7%$ and the binding efficiency in case of not using it was $87{\pm}18%$ (p>0.05). There was no significant difference between them. In binding efficiency according to kinds of anticoagulants, ACD was $98{\pm}4%$, CPDA was $97{\pm}6%$ and heparin was $89{\pm}20%$ (p<0.001). Conclusion: When modified in vivo erythrocyte labeling technique is used with Tc-99m, the methods to obtain the highest labeling efficiency are as follow. The withdrawing blood volume should be over 3 mL, tinning time should be kept between 20 min and 35 min, and incubation time should be kept between 10 min and 40 min. ACD or CPDA have to be used as a anticoagulant except heparin and the blood samples should be placed on rotating invertor during incubation.
Purpose: Cancer specific killing can be achieved by therapeutic gene activated by cancer specific promotor. Expression of sodium iodide symporter (NIS) gene causes transportation and concentration of iodide into the cell, therefore radioiodine treatment after NIS gene transfer to cancer cell could be a form of radionuclide gene therapy. luciferase (Luc) gene transfected cancer cell can be monitored by in vivo optical imaging after D-luciferin injection. Aims of the study are to make vector with both therapeutic NIS gene driven by AFP promoter and reporter Luc gene driven by CMV promoter, to perform hepatocellular carcinoma specific radiodiodine gene therapy by the vector, and assessment of the therapy effect by optical imaging using luciferase expression. Materials and Methods: A Vector with AFP promoter driven NIS gene and CMV promoter driven Luc gene (AFP-NIS-CMV-Luc) was constructed. Liver cancer cell (HepG2, Huh-7) and non liver cancer cell (HCT-15) were transfected with the vector using liposome. Expression of the NIS gene at mRNA level was elucidated by RT-PCR. Radioiodide uptake, perchlorate blockade, and washout tests were performed and bioluminescence also measured by luminometer in these cells. In vitro clonogenic assay with 1-131 was performed. In vivo nuclear imaging was obtained with gamma camera after 1-131 intraperitoneal injection. Results: A Vector with AFP-NIS-CMV-Luc was constructed and successfully transfected into HepG2, Huh-7 and HCT-15 cells. HepG2 and Huh-7 cells with AFP-NIS-CMV-Luc gene showed higher iodide uptake than non transfected cells and the higher iodide uptake was totally blocked by addition of perchlorate. HCT-15 cell did not showed any change of iodide uptake by the gene transfection. Transfected cells had higher light output than control cells. In vitro clonogenic assay, transfected HepG2 and Huh-7 cells showed lower colony count than non transfected HepG2 and Huh-7 cells, but transfected HCT-15 cell did not showed any difference than non transfected HCT-15 cell. Number of Huh-7 cells with AFP-NIS-CMV-Luc gene transfection was positively correlated with radioidine accumulation and luciferase activity. In vivo nuclear imaging with 1-131 was successful in AFP-NIS-CMV-Luc gene transfected Huh-7 cell xenograft on nude mouse. Conclusion: A Vector with AFP promoter driven NIS and CMV promoter driven Luc gene was constructed. Transfection of the vector showed liver cancer cell specific enhancement of 1-131 cytotoxicity by AFP promoter, and the effect of the radioiodine therapy can be successfully assessed by non-invasive luminescence measurement.
Background: Reoperations on the aortic root or the ascending aorta are being performed with increasing frequency and remain a challenging problem. This study was performed to analyze the results of reoperations on the ascending aorta and aortic root. Material and Method: Between May 1995 and April 2001, 30 patients had reoperations on the ascending aorta and aortic root and were reviewed retrospectively. The mean interval between the previous repair and the actual reoperation was 56 months(range 3 to 142 months). Seven patients(23.3%) had two or more previous operations. The indications for reoperations were true aneurysm in 7 patients(23.3%), prosthetic valve endocarditis in 6(20%), false aneurysm in 5(16.7%), paravalvular leak associated with Behcet's disease in 4(13.3%), malfunction of prosthetic aortic valve in 4(13.3%), aortic dissection in 3(10%), and annuloaortic ectasia in 1(3.3%). The principal reoperations performed were aortic root replacement in 17 patients(56.7%), replacement of the ascending aorta in 8(26.7%), aortic and mitral valve replacement with reconstruction of fibrous trigone in 2(6.6%), patch aortoplasty in 2(6.6%), and aortic valve replacement after Bentall operation in 1 (3.3%). The cardiopulmonary bypass was started before sternotomy in 7 patients and the hypothermic circulatory arrest was used in 16(53.3%). The mean time of circulatory arrest, total bypass, and aortic crossclamp were 20$\pm$ 12 minutes, 228$\pm$56 minutes, and 143$\pm$62 minutes, respectively Result: There were three early deaths(10%). The postoperative complications were reoperation for bleeding in 7 patients(23.3%), cardiac complications in 5(16.7%), transient acute renal failure in 2(6.6%), transient focal seizure in 2(6.6%), and the others in 5. The mean follow-up was 22.8 $\pm$20.5 months. There were two late deaths(7.4%). The actuarial survival was 92.6$\pm$5.0% at 6 years. One patient required reoperation for complication of reoperation on the ascending aorta and aortic root(3.7%). The 1- and 6-year actuarial freedom from reoperation was 100% and 83.3$\pm$15.2%, respectively. One patient with Behcet's disease are waiting for reoperation due to false aneurysm, which developed after aortic root replacement with homograft. There were no thromboembolisms or anticoagulant related complications. Conclusions: This study suggests that reoperations on the ascending aorta and aortic root can be performed with acceptable early mortality and morbidity, and adequate surgical strategies according to the pathologi conditions are critical to the prevention of the reoperation.
This study was carried out to estimate the selection of appropriate forage crops, proper application levels of livestock manure, and carrying capacity per unit area for organic livestock, as influenced by livestock manure application levels compared with chemical fertilizer to corn and sorghum $\times$ sorghum hybrid, in order to produce organic forages by utilizing livestock manure. For both corns and sorghum $\times$ sorghum hybrids, no fertilizer plots had significantly (p<0.05) lower annual dry matter (DM), crude protein (CP) and total digestible nutrients (TDN) yields than those of other plots, whereas the N+P+K plots ranked the highest yields, followed by 150% cattle manure plots and 100% cattle manure plots. Dry matter, CP and TDN yields of cattle manure plots were significantly (p<0.05) higher than those of no fertilizer and P+K plots. In applying cattle manure, the yields of cattle slurry plots tended to be a little higher than those of composted cattle manure plots. Assuming that corns and sorghum $\times$ sorghum hybrids produced from this trial were fed at 70% level to 450kg of Hanwoo heifer with 400g of average daily gain, livestock carrying capacity (head/year/ha) ranked the highest in N+P+K plots of the case of corns (mean 6.7 heads), followed by 150% cattle slurry plots (mean 5.6 heads), 150% composted cattle manure plots (mean 4.8 heads), 100% cattle slurry plots (mean 4.4 heads), 100% composted cattle manure plots (mean 4.3 heads), P+K plots (mean 4.1 heads), and no fertilizer plots (mean 3.1 heads). Meanwhile, in case of sorghum $\times$ sorghum hybrids, N+P+K plots (mean 5.7 heads) ranked the highest carrying capacity, followed by $100{\sim}150%$ cattle slurry plots (mean $4.8{\sim}5.2$ heads), 150% composted cattle manure plots (mean 4.7 heads), 100 % composted cattle manure plots (mean 4.3 heads), P+K plots (mean 3.8 heads), and no fertilizer plots (mean 3.4 heads). The results indicated that replacing chemical fertilizer by livestock manure application to cultivation soil for forage crops could enhance not only DM and TDN yields, but also organic stock carrying capacity. In conclusion, it was conceived that organic forage production by reutilizing livestock manure might contribute to reduced environmental pollution and the production of environment friendly agricultural products through resources recycling.
An, Chang Hyeok;Lim, Sung Yong;Suh, Gee Young;Park, Gye Young;Park, Jung Woong;Jeong, Seong Hwan;Lim, Si Young;Oui, Misook;Koh, Won-Jung;Chung, Man Pyo;Kim, Hojoong;Kwon, O Jung
Tuberculosis and Respiratory Diseases
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v.54
no.1
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pp.71-79
/
2003
Background : A bronchoalveolar lavage(BAL) is useful in diagnosing the etiology of bilateral pulmonary infiltrations, but may worsen the oxygenation and clinical status in severely hypoxemic patients. This study assessed the safety and efficacy of the continuous positive airway pressure(CPAP) using a conventional mechanical ventilator via a face mask as a tool for maintaining the oxygenation level during BAL. Methods : Seven consecutive patients with the bilateral pulmonary infiltrates and severe hypoxemia ($PaO_2/FIO_2$ ratio ${\leq}200$ on oxygen 10 L/min via mask with reservoir bag) were enrolled. The CPAP 5-6 $cmH_2O(F_IO_2\;1.0)$ was delivered through an inflatable face mask using a conventional mechanical ventilator. The CPAP began 10 min before starting the BAL and continued for 30 min after the procedure was completed. A bronchoscope was passed through a T-adapter and advanced through the mouth. BAL was performed using the conventional method. The vital signs, pulse oxymetry values, and arterial blood gases were monitored during the study. Results : (1) Median age was 56 years(male:female=4:3). (2) The baseline $PaO_2$ was $78{\pm}16mmHg$, which increased significantly to $269{\pm}116mmHg$(p=0.018) with CPAP. After the BAL, the $PaO_2$ did not decrease significantly but returned to the baseline level after the CPAP was discontinued. The $SpO_2$ showed a similar trend with the $PaO_2$ and did not decrease to below 90 % during the duration of the study. (3) The $PaCO_2$ increased and the pH decreased significantly after the BAL but returned to the baseline level within 30 min after the BAL. (5) No complications directly related to the BAL procedure were encountered. However, intubation was necessary in 3 patients(43 %) due to the progression of the underlying diseases. Conclusion : In severe hypoxemic patients, CPAP using a face mask and conventional mechanical ventilator during a BAL might allow minimal alterations in oxygenation and prevent subsequent respiratory failure.
Kim, Eun Ji;Kim, Hae Sook;Hur, Man Hoe;Lee, Sang Geel
Clinical and Experimental Pediatrics
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v.45
no.10
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pp.1204-1212
/
2002
Purpose : Early surfactant therapy with either gentle ventilation, high-frequency ventilation or aggressive weaning of mechanical ventilation are principles for the treatment of respiratory distress syndrome(RDS). We studied to determine the accessibility of noninvasive nasal continuous positive airway pressure(CPAP) rather than mechanical ventilation by invasive intubation after early surfactant therapy. Methods : The study group consisted of 14 infants who were born and diagnosed with moderate respiratory distress syndrome and received early surfactant therapy with nasal CPAP of PEEP 5-6 cm $H_2O$ within two hours after birth in the Fatima neonatal intensive care unit for two years from January 1999 to August 2001. The control group consisted of 15 infants who were diagnosed with the disease and could be weaned from mechanical ventilator within five days after birth during the same period. Results : The characteristics, the severity of clinical symptoms and laboratory findings in the two groups at birth showed no significant difference. Neither did the interim analysis of laboratory data in two groups. Of 14 infants in the study group who received nasal CPAP after early surfactant therapy, only two infants showed weaning failure with this therapy. In the response cases, duration of CPAP was five days and mean airway pressure was $5.4{\pm}0.5cm$$H_2O$. Two had the complication of CPAP with abdominal distension. Final complications and outcomes in the two groups showed no signifcant difference(P>0.05). Conclusion : The clinical courses in the two groups showed no significant difference. Therefore, we suggest that early surfactant therapy with noninvasive nasal CPAP is a simple and safe method rather than aggressive weaning after invasive mechanical ventilation in moderate respiratory distress syndrome.
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