• Journal of The Korean Radiological Technologist Association
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• v.30 no.1
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• pp.63-69
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• 2004

Purpose : We underwent this study to evaluate the factors which influence labelling efficiency when modified in vivo erythrocyte labeling technique was used. Materials and methods : Thirty healthy volunteers (M : F = 19 : 11, age : 25$\pm$2yrs)

• The Korean Journal of Nuclear Medicine
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• v.38 no.4
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• pp.300-305
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• 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 After suitable tinning time (5 min, 20 min, 35 min) passed by, blood (5 mL, 3 mL or 1 mL) was withdrawn into 10 mL syringe previously containing Tc-99m (740 MBq) and anticoagulant (heparin, ACD or CPDA) through 19-gauged scalp needle.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.

• The Korean Journal of Nuclear Medicine
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• v.20 no.1
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• pp.39-43
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• 1986

[ $^{99m}Technetium-Heat$ ] damaged erythrocyte were used as spleen scanning agents in 12 patients from July, 1985 to April, 1986. We used this scan to evaluate situs inversus, asplenia, accessory spleen, hypersplenism, splenic infarction, tumor staging and evaluation of therapy, especially when the $^{99m}Tc-tin$ colloid scans were not definite for diagnosis. The techniques applied to these scans were in vivo/in vitro-labeling method and heating-method to damage the erythrocytes. Liver-to-spleen uptake ratios were increased upto 100 : 1 and interference from the left lobe of the liver was eliminated. These scans were helpful to evaluate the spleen.

• Journal of Applied Biological Chemistry
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• v.56 no.4
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• pp.191-197
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• 2013

To classify the chemical hazard according to globally harmonized system of classification and labeling of chemicals (GHS), we investigated the genotoxicity of three chemicals, methyl myristate, 2-ethylhexanoic acid zinc salt, N,N,N',N'-tetrakis(2-hydroxyethyl) ethylenediamine, using male ICR mice bone marrow cells for the screening of micronucleus induction. Although these three chemicals have already been tested numerous times, a micronucleus test has not been conducted. The seven week-old male ICR mice were tested at three dosages for the three chemicals, respectively. After 24 h of oral administration with the three chemicals, the mice were sacrificed and their bone marrow cells were prepared for smearing slides. As a result of counting the micronucleated polychromatic erythrocyte (MNPCE) of 2,000 polychromatic erythrocytes, all treated groups expressed no statistically significant increase of MNPCE compared to the negative control group. There were no clinical signs related with the oral exposure of these three chemicals. It was concluded that these three chemicals did not induce micronucleus in the bone marrow cells of ICR mice, and there was no direct proportion with dosage. These results indicate that the three chemicals have no mutagenic potential under each test condition, and it is not classified these chemicals as mutagens by GHS.

• Journal of radiological science and technology
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• v.30 no.3
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• pp.259-263
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• 2007

For the preparation of $^{99m}Tc$-labeled RBC, $10{\sim}20\;{\mu}g/kg$ of Stannous(II) chloride and $10{\sim}40\;min$ of preparation was used. For finding out the effect of contrast agent, the blood samples were collected in three days, seven days, and 1 months after the diagnostic procedure. In the normal volunteer, the concentration of reducing agent and preparation time did not effect on the radiochemical yield. But in the patients, 10 mg of Stannous(II) chloride and 60 min incubation times was shown high radiochemical yield. Contrast agent has a significant effect on the radiochemical yield. Although the blood samples which were collected after seven days of diagnostic procedure did not effect on the radiochemical yield of $^{99m}Tc$-labeled RBC, but the radiochemical yield of $^{99m}Tc$-labeled RBC which was prepared with a sample of high concentration of contrast agent in blood led to low radiochemical yield. For these samples, the modified method showed high radiochemical yield than previous in vivo preparation method. The recommended method is followed. Blood collecting was performed at 30 minutes after injection of reducing agent, and it is centrifuged for removal of plasma. After addition of $^{99m}TcO^-_4$, sample reservoir was rotated. After addition of normal saline, and it is centrifuged for separation of saline. Then $^{99m}Tc$-labeled RBC was obtained after removal of saline.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.681-692
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• 1996

The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.