Gaja, A.;Meng, C.L.;Sato, M.;Nakajima, T.;Kubota, Chikara;Kojima, T.
Asian-Australasian Journal of Animal Sciences
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v.23
no.1
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pp.25-32
/
2010
The present study aimed to examine the effects of ${\gamma}$-linolenic acid (GLA) supplementation to in vitro culture (IVC) medium on in vitro developmental competence, freezability and morphology of in vitro matured and fertilized bovine embryos. In vitro produced (IVP) bovine zygotes were cultured in IVC medium supplemented with 0 (negative control), 15, 31, 62, 125, 250, 500 or 1,000 ppm GLA, 250 ppm linoleic acid albumin (LAA) and without any supplement as a control. Day 6 blastocysts derived from culture control were cultured in IVC medium containing either 62, 250 GLA or 250 LAA for 24 h, and at Day 7 were subjected to freezing or morphological examination by electron microscope. GLA 15 showed a tendency to have a higher cleavage rate at Day 2 (70.3%) than other groups. The hatching rate at Day 9 in LAA (38.2%) was significantly higher than the control and all treatment groups (p<0.05), while the blastocyst rate in LAA (32.4%) did not differ from those of 15 (30.5%), 31 (27.1%), and 62 GLA (33.1%) or the control (35.1%). GLA in concentrations of 125, 250, 500, and 1,000 ppm had significantly detrimental effect on the blastocyst rate compared to 15, 31 and 62 ppm GLA, LAA, and control groups (p<0.05). In contrast, the highest post-thaw survival rate (100%) was observed in the control group (p<0.01). Large lipid droplets were observed in the cytoplasm of trophoblastic cells, even in the control, but were abundant in GLA groups. Taking the results of the study into consideration, the addition of GLA to the culture medium for IVP bovine embryos at the dose of 15 ppm increased the developmental competence of zygotes and enhanced the cleavage rate up to Day 2. However, blastulation rate and post-thaw survival were not increased when GLA was added to the culture media.
Aliphatic alkenals having 6 to 13 carbons were evaluated for antifungal activity against Saccharomyces cerevisiae. The activity was gradually increased with chain length, e.g., (E)-2-decenal and (E)-2-undecenal exhibited maximum potency, while (E)-2-dodecenal and (E)-2-tridecenal were completely inactive. Alkenals showed increasing inhibitory activity with chain length, as in the case of antifungal activity, towards glucose-induced medium acidification by the plasma membrane $H^+$-ATPase of S. cerevisiae. The group including (E)-2-nonenal, (E)-2-decenal, and (E)-2-undecenal exhibited maximum potency, but the potency of (E)-2-dodecenal and (E)-2-tridecenal demonstrated a sudden drop with respect to the former group. (E)-2-Nonenal revealed dose-responsive inhibition to the medium acidification and inhibited over 90% at a concentration of 1.25 mM ($175.3{\mu}g$/ml). In contrast to (E)-2-undecenal whose inhibitory efficiency increased with incubation time, inhibition by (E)-2-dodecenal was reversed with time. Of the tested alkenals, (E)-2-heptenal and (E)-2-octenal most highly inhibited ATP hydrolytic activity by the plasma membrane $H^+$ ATPase, while (E)-2-heptenal at 10 mM ($1121.8{\mu}g$/ml) showed an inhibitory efficacy of 93.2%.
Targeting the cystine/glutamate exchange transporter, system xc-, is a promising anticancer strategy that induces ferroptosis, which is a distinct form of cell death mediated by iron-dependent lipid peroxidation. The concentration of ⳑ-cystine in culture medium is higher than the physiological level. This study was aimed to evaluate the effects of ⳑ-cystine concentration on the efficacy of ferroptosis inducers in hepatocellular carcinoma cells. This study showed that treatment with sulfasalazine or erastin, a system xc- inhibitor, decreased the viability of Huh6 and Huh7 cells in a dose-dependent manner, and the degree of growth inhibition was greater in medium containing a physiological ⳑ-cystine concentration of 83 µM than in commercial medium with a concentration of 200 µM ⳑ-cystine. However, RSL3, a glutathione peroxidase 4 inhibitor, decreased cell viability to a similar extent in media containing both ⳑ-cystine concentrations. Sulfasalazine and erastin significantly increased the percentages of propidium iodide-positive cells in media with 83 µM ⳑ-cystine, but not in media with 200 µM ⳑ-cystine. Sulfasalazine- or erastin-induced accumulation of lipid peroxidation as monitored by C11-BODIPY probe was higher in media with 83 µM ⳑ-cystine than in media with 200 µM ⳑ-cystine. In contrast, the changes in the percentages of propidium iodide-positive cells and lipid peroxidation by RSL3 were similar in both media. These results showed that sulfasalazine and erastin, but not RSL3, were efficacious under conditions of physiological ⳑ-cystine concentration, suggesting that medium conditions would be crucial for the design of a bioassay for system xc- inhibitors.
Kim, Ki-Jeong;Jung, Hong-Ryang;Lim, Cheong-Hwan;Hong, Dong-Hee;Shim, Jae-Goo;You, In-Gyu
Journal of Digital Convergence
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v.12
no.3
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pp.353-358
/
2014
This study was conducted to analyze the patient's exposed dose targeting the patients who had acute ischemic stroke symptoms and CT brain perfusion scan, by comparing fixed time technique and bolus tracking technique which was provided by the manufacturer and to identify the Time graph to implement the usability of contrast medium's tracking technique the best contrast enhancement intervals. $CTDI_{VOL}$ of PCT in patient appeared to be 431.72mGy in fixed scan delay protocol, whereas 323.61mGy in Bolus tracking technique. The value of DLP appeared to be $1243.47mGy{\cdot}cm$ in fixed scan delay protocol, whereas $932mGy{\cdot}cm$ in Bolus tracking technique. Time graph appeared to be various in fixed scan delay protocol, whereas the optimal time graph could be obtained in Bolus tracking. The exposure dose could be reduced by 25% applying Bolus tracking technique when taking brain perfusion CT scan.
To assay the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 13 boars(Duroc. Landrace and Yorkshire) which had been proved to be fertile in the past. then, were preserved in BWW medium or in raw state at 18$^{\circ}C$ or at room temperature. The preserved semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding). ova penetrated by sperm(penetration) and formation of a male pronucleus(pronucleus formation) and also numbers of both bound and penetrated sperm per ovum. Between BWW and TBM medium for boar sperm. no difference in the results of hamster test was obtained. The boar spermatozoa in BWW medium, BWW with caffeine, BWW with heparin, and BWW with both caffeine and heparin showed no difference in the results of hamster test. The boar spermatozoa in BWW medium containing both calcium and RSA showed considerably higher rates of sperm binding, penetration and pronucleus formation as well as higher numbers of both bound and penetrated sperm than those not containing calcium with or without BSA( p<0.01) and also the same results higher than that containing calcium without BSA( p< 0.05). The boar spermatozoa irradiated by X-ray(70 KVP, 20mA) for 3 seconds. then, maintained at 18$^{\circ}C$ for 18 hours showed considerably lower rate of sperm binding than all the other groups including the control and X-ray groups irradiated by smaller dose or maintained for shorter period(p<0.01), and also showed lower number of bound sperm than the other groups(p<0.01, p<0.05). All the control groups of both raw and diluted sperm in BWM medium showed higher rates of sperm binding, penetration and pronucleus formation as well as higher number of penetrated sperm than all the X-ray groups irradiated for 3 seconds(70KVP, 20mA) and maintained for either 3 or 18 hours (p<0.01, p<0.05). At the same time the control groups of diluted sperm showed considerably higher rates of sperm penetration and pronucleus formation than the control group of raw sperm( p<0.01). These results indicates that fertile boar sperm showed considerably lower rates In the results of hamster test, when incubated in the medium without calcium and irradiated by X-ray than when incubated in the medium with calcium and not irradiated by X-ray, respectively, to prove consequently that hamster test would be of great value in assaying the fertilizing capacity of boar spermatozoa.
The objective of this study was to detect the fragments generated during IV (intravenous) catheter injection of contrast medium and drug administration in a clinical setting and removal was performed by experimentally producing a phantom, and to compare the radiography, ultrasonography, and multi-detector computed tomography (MDCT) imaging and radiation dose. A 1 cm fragment of an 18 gage Teflon$^{(R)}$ IV catheter with saline was inserted into the IV control line. Radiography, CT, and ultrasonography were performed and radiography and CT dose were calculated. CT and ultrasonography showed an IV catheter fragment clinically and radiography showed no visible difference in the ability to provide a useful image of an IV catheter fragment modality (p >.05). Radiography of effective dose ($0.2139mSv{\cdot}Gy^{-1}{\cdot}cm^{-2}$) form DAP DAP ($0.93{\mu}Gy{\cdot}m^2 $), and dose length product (DLP) ($201mGy{\cdot}cm$) to effective dose was calculated as 0.483 mSv. IV catheter fragment were detected of radiography, ultrasonography and CT. These results can be obtained by menas of an excellent IV catheter fragment of detection capability CT. However, CT is followed by radiation exposure. IV catheter fragment confirming the position and information recommend an ultrasonography.
Objective: To separate/enrich tumor stem-like cells from the human esophageal carcinoma cell line OE-19 by using serum-free suspension culture and to identify their biological characteristics and radiation resistance. Methods: OE-19 cells were cultivated using adherent and suspension culture methods. The tumor stem-like phenotype of CD44 expression was detected using flow cytometry. We examined growth characteristics, cloning capacity in soft agar, and radiation resistance of 2 groups of cells. Results: Suspended cells in serum-free medium formed spheres that were enriched for CD44 expression. CD44 was expressed in 62.5% of suspended cells, but only in 11.7% of adherent cells. The suspended cells had greater capacity for proliferation and colony formation in soft agar than the adherent cells. When the suspended and adherent cells were irradiated at 5 Gy, 10 Gy, or 15 Gy, the proportion of CD44+ suspended cells strongly and weakly positive for CD44 was 77.8%, 66.5%, 57.5%; and 21.7%, 31.6%, 41.4%, respectively. In contrast, the proportion of CD44+ adherent cells strongly positive for CD44 was 18.9%, 14.%, and 9.95%, respectively. When the irradiation dose was increased to 30 Gy, the survival of the suspended and adherent cells was significantly reduced, and viable CD44+ cells were not detected. Conclusion: Suspended cell spheres generated from OE-19 esophageal carcinoma cells in serum-free stem medium are enriched in tumor stem-like cells. CD44 may be a marker for these cells.
Background: To investigate the inhibitory effect and the underlying mechanism of triptolide on cultured human endometrial carcinoma HEC-1B cells and corresponding xenograft. Materials and Methods: For in vitro studies, the inhibition effect of proliferation on HEC-1B cell by triptolide was determined by MTT assay; cell cycle and apoptosis of the triptolide-treated and untreated cells were detected by flow cytometry. For in vivo studies, a xenograft tumor model of human endometrial carcinoma was established using HEC-1B cells, then the tumor-bearing mice were treated with high, medium, and low-dose ($8{\mu}g$, $4{\mu}g$ and $2{\mu}g/day$) triptolide or cisplatin at $40{\mu}g/day$ or normal saline as control. The mice were treated for 10-15 days, during which body weight of the mice and volume of the xenograft were weighted. Then expression of Bcl-2 and vascular endothelial growth factor (VEGF) was analyzed by SABC immunohistochemistry. Results: Cell growth was significantly inhibited by triptolide as observed by an inverted phase contrast microscope; the results of MTT assay indicated that triptolide inhibits HEC-1B cell proliferation in a dose and time-dependent manner; flow cytometry showed that low concentration (5 ng/ml) of triptolide induces cell cycle arrest of HEC-1B cells mainly at S phase, while higher concentration (40 or 80 ng/ml) induced cell cycle arrest of HEC-1B cells mainly at G2/M phase, and apoptosis of the cells was also induced. High-dose triptolide showed a similar tumor-inhibitory effect as cisplatin (-50%); high-dose triptolide significantly inhibited Bcl-2 and VEGF expression in the xenograft model compared to normal saline control (P<0.05). Conclusions: triptolide inhibits HEC-1B cell growth both in vitro and in mouse xenograft model. Cell cycle of the tumor cells was arrested at S and G2/M phase, and the mechanism may involve induction of tumor cell apoptosis and inhibition of tumor angiogenesis.
Kim, Jung Yul;Kim, Joo Yeon;Nam-Koong, Hyuk;Kang, Chun Goo;Kim, Jae Sam
The Korean Journal of Nuclear Medicine Technology
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v.18
no.1
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pp.33-42
/
2014
Purpose: I-131 scan using High Energy (HE) collimator is generally used. While, Medium Energy (ME) collimator is not suggested to use in result of an excessive septal penetration effects, it is used to improve the sensitivities of count rate on lower dose of I-131. This research aims to evaluate I-131 SPECT/CT image quality using by HE and ME collimator and also find out the possibility of ME collimator clinical application. Materials and Methods: ME and HE collimator are substituted as Siemens symbia T16 SPECT/CT, using I-131 point source and NEMA NU-2 IQ phantom. Single Energy Window (SEW) and Triple Energy Windows (TEW) are applied for image acquisition and images with CTAC and Scatter correction application or not, applied different number of iteration and sub set are reconstructed by IR method, flash 3D. By analysis of acquired image, the comparison on sensitivities, contrast, noise and aspect ratio of two collimators are able to be evaluated. Results: ME Collimator is ahead of HE collimator in terms of sensitivity (ME collimator: 188.18 cps/MBq, HE collimator: 46.31 cps/MBq). For contrast, reconstruction image used by HE collimator with TEW, 16 subset 8 iteration applied CTAC is shown the highest contrast (TCQI=190.64). In same condition, ME collimator has lower contrast than HE collimator (TCQI=66.05). The lowest aspect ratio for ME collimator and HE collimator are 1.065 with SEW, CTAC (+) and 1.024 with TEW, CTAC (+) respectively. Conclusion: Selecting a proper collimator is important factor for image quality. This research finding tells that HE collimator, which is generally used for I-131 scan emitted high energy ${\gamma}$-ray is the most recommendable collimator for image quality. However, ME collimator is also applicable in condition of lower dose, lower sensitive if utilizing energy window, matrix size, IR parameter, CTAC and scatter correction appropriately.
This study compares the difference between the remaining amount in syringes according to injection method and the dose of physiological saline when the radiopharmaceutical 18F-FDG isotope is injected into patients who visited the hospital for PET examination. After performing a CT or MRI using a contrast medium when 18F-FDG was injected into 40 patients who came to the hospital for PET examination without removing the 3-way, the radioactivity remaining in the syringe and the 3-way was measured and the dose of radioactivity confirmed. At this time, 20 patients were divided into different dose groups of physiological saline. Another injection method was used to compare the remaining amount of the syringe and the difference in the remaining amount according to the amount of physiological saline when the injection was performed with an extension. After an injection of 18F-FDG, the actual administered dose was confirmed by measuring the radioactivity remaining in the syringe and the 3-way or extension with a calibrator. As a result of measuring the radioactivity of the syringe before administering the radiopharmaceutical and the radioactivity of the syringe and the 3-way or extension after administration, the 3-way injection method in 10 cc's of physiological saline had the lowest residual amount. The remaining amount increased in the order of the extension to 10 cc's of saline and the 3-way to 5 cc's of saline. Likewise, the 5 cc's saline solution and the extension injection method demonstrated the highest residual amounts. The residual difference with the lowest level of remaining injection method was found to be 0.053 mCi. It was found that in a PET examination, by considering the radioactivity remaining in the 3-way and extension and by adjusting the dose of physiological saline, if the intended dose to be administered to the actual patient is determined, it is possible to administer the radiopharmaceutical to the patient more accurately.
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