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Development and Expression of Porcine Embryos by Direct Injection of Sperm Treated with Exogenous DNA (외래유전자 도입정자를 이용한 돼지 체외성숙 난포란의 Intracytoplasmic Sperm Injection (ICSI) 후 후기 배로의 발달율과 외래유전자의 발현에 관한 연구)

  • 정기화;조성근
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.45-53
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    • 2002
  • The main goal of this study was to produce transgenic porcine embryos by direct injection of sperm-mediated exogenous DNA. Spermatozoa (6$\times$10$^{6}$ sperms of final concentration) were mixed with pcDNA LAC Z (20 ng/$\mu$l) and subjected into electroporation (300~750 volts, 25 $\mu$F, 0.4 cm electrode). After sperm injection, the oocytes were activated electrically (1.7 KV/cm, 30$\mu$sec, single pulse) in 0.3 M mannitol solution or not. The sperm injected eggs were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air fur 144 h. The rates of cleavage and development into blastocyst stage in activation group were significantly higher than those of non-activation group (79.6% and 24.1% vs. 46.3% and 14.4%, respectively, p<0.05). Control oocytes and shame injection were developed to blastocysts low (2.5%). Sixty five (27.1%) out of 240 embryos observed in activation and non-activation groups were showed positive by X-gal staining. However, all embryos in both groups were expressed partial or mosaic pattern. These results suggested that electrical stimulation far oocytes activation after sperm injection enhances the incidence of both fertilization and development fellowing sperm injection in the pig. Our study also suggested that sperm-mediated transfer of exogenous DNA by ICSI would be used as a valuable tool for the production of transgenic porcine embryos.

Biodistribution and Hepatic Metabolism of Galactosylated $^{111}In-Antibody-Chelator$ Conjugates: Comparison with $^{111}In-Antibody-Chelator$ Conjugates ($^{111}In$-표지 갈락토즈 접합 항체의 체내분포 및 간에서의 대사 : $^{111}In$-표지 항체와의 비교연구)

  • Kwak, Dong-Suk;Jeong, Kyu-Sik;Ha, Jeoung-Hee;Ahn, Byeong-Cheol;Lee, Kyu-Bo;Paik, Chang-H.;Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.6
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    • pp.402-417
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    • 2003
  • Purpose: To evaluate the use of monoclonal antibody (MoAb) as a carrier of the receptor-binding ligand the receptor mediated uptake into liver and subsequent metabolism of $^{111}In-labeled$ galactosylated MoAb-chelator conjugates were investigated and compared with those of $^{111}In$ labeled MoAb. Materials and Methods : T101 MoAb, $IgG_2$ against human lymphocytic leukemic cell, conjugated with cyclic DTPA dianhydride (DTPA) or 2-p-isothiocyanatobenzyl-6-methyl-DTPA (1B4M) was galactosylated with 2-imino-2-methoxyethyl-1-thio-${\beta}$-D-galactose and then radiolabeled with $^{111}In$. Biodistribution and metabolism study was peformed with two $^{111}In-conjugates$ in mice and rats. Results: $^{111}In-labeled$ T101 and its galactosylated conjugates were taken to the liver by the time, mostly within 10 min. However DTPA conjugate was retained longer in the liver than the 1B4M conjugate (55% vs 20% of injected dose at 44 hr). During this time, the radiornetabolite of DTPA conjugate was excreted similarly into urine (24%) and feces (17%). The radiometabolite of 1B4M was excreted primarily into feces (68%) rather than urine (8%). Size exclusion HPLC analysis of the bile and supernatant of liver homogenate showed two peaks the first (35%) with the retention time (Rt) identical to IgG and the second (65%) with Rt similar to free $^{111}In$ at 3 hr post-injection for the 1B4M conjugate, indicating that the metabolite is rapidly excreted through the biliary system. in contrast to DTPA conjugate, the small $^{111}In-DTPA-like$ metabolite was the major radioindium component (90%) in the liver homogenate as early as 3 hour post-injection, but the cumulative radioindium activity in feces was only 17% at 44 hour, indicating that the metabolite from DTPA conjugate does not clear readily through the biliary tract. Conclusion: The galactosylation of the MoAb conjugates resulted in higher hepatocyte uptake and enhanced metabolism, compared to those without galactosylation. Metabolism of the MoAb-conjugates is different between compounds radiolabled with different chelators due to different characteristics of radiometabolites generated in the liver.

Effect of Sucrose Concentration on Survival After Frozen-thawed of Bovine IVF Blastocysts in Ethylene Glycol Based Freezing Medium for Slow-Cooling (소 체외수정란의 Slow Freezing을 위해서 Ethylene Glycol 동결보호제에 Sucrose 첨가 농도에 의한 동결효율)

  • 조상래;김현종;최창용;진현주;손동수;최선호
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.797-804
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    • 2006
  • The present study was undertaken to investigate the post-thawed survivability of bovine embryo depending on different dose of ethylene glycol and sucrose. Ovaries were collected at local slaughterhouse and the cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured at 39°C in an atmosphere of 5% CO2 incubator. For conventional slow-freezing, d 7 or 8 expanded blastocysts were collected. Embryos were equilibrated in 1.5 M and 1.8 M ethylene glycol(EG) with 0.1 M and 0.3 M sucrose in Dulbecco's phosphate-buffered saline(D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25ml-straw and placed directly into cooling chamber of programmable freezer precooled to 󰠏7°C, after 2 min, the straw was seeded, maintained at 󰠏7°C for 8 min, and then cooled to 󰠏35°C at 0.3°C/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 sec and exposed to 37°C water for 20 sec. Straws were then removed from 37°C water. Rates of blastocyst survive and hatching were evaluated at 24 to 72 h post-warming. No difference of the survivability was shown between 1.5 M and 1.8 M EG (71 and 70%, respectively). Addition of 0.1 M sucrose to 1.5 M and 1.8 M ethylene glycol in the freezing solution did not differ significantly embryo survival (74 and 77%, respectively), whereas survival rates was higher(89%) in freezing solution contained 0.3M sucrose to 1.8M EG compared with 0.3M sucrose to 1.5M EG group(71%). However, there was no difference in the overall total cell number between the two groups (122±1.8 vs 131±1.4, respectively). In conclusion, the results suggest that 0.3 M sucrose in 1.8 M EG may be optimal condition for freezing and thawing methods with in vitro produced embryos and may be applied to on-farm conditions for embryo transfer.

Gene Expression of Candidate Genes Involved in Fat Metabolism During In vitro Adipogenic Differentiation of Bovine Mesenchymal Stem Cell (Bovine Mesenchymal Stem Cell의 지방분화를 이용한 지방대사관련 후보 유전자의 발현분석)

  • Kim, Sung-Kon;Kim, Nam-Kuk;Yoon, Du-Hak;Kim, Tae-Hun;Yang, Boo-Keun;Lee, Hyun-Jeong
    • Journal of Animal Science and Technology
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    • v.52 no.4
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    • pp.265-270
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    • 2010
  • Adipogenesis has been one of the most intensely studied models of cellular differentiation. During adipogenesis, differential expression of many adipogenesis related genes lead to profound changes in cellular, morphological, and physiological characteristics of the differentiating cells. The aim of the present study was to examine the expression levels of adipogenic candidate genes, cAMP early repressor (ICER), nephroblastoma over-expressed protein (NOV), heat shock protein beta 1 (HSPB1) and succinate dehydrogenase (SDH), during adipogenesis of bovine mesenchymal stem cells (BMSC). The BMSC were cultured in DMEM / low glucose medium with adipogenic inducers for 6 days and the expression of various candidate genes which seemed related to adipogenesis were measured by real-time PCR. This study showed that the expression of peroxisome proliferator activated receptor ${\gamma}$(PPAR${\gamma}$) and fatty acid binding protein 4 (FABP4) genes as adipogenic indicators were increased to 3.11 and 3.11 folds on day 6 than on day 0, respectively (p<0.05). To determine whether candidate genes were related to adipogenesis, the expression levels of ICER, NOV, HSPB1, and SDH genes were measured during adipogenesis in BMSC. Our results showed that the expression level of ICER gene was significantly increased to 4.12 folds (0.01729 vs. 0.07138; p<0.05), whereas NOV, HSPB1, and SDH genes were decreased to 2.89, 3.18 and 2.36 folds, respectively, on day 6 when compared to day 0. These results suggest that these candidate genes have stimulatory or inhibitory effects on adipogenesis in BMSC, indicating that these genes may be directly or indirectly related to the adipogenic event of adipose precursor cells.

A Dosimetric Evaluation of Large Pendulous Breast Irradiation in Prone Position (Large Pendulous Breast 환자의 방사선 치료에 있어서 엎드린 자세의 유용성 평가)

  • Hong, Chae-Seon;Ju, Sang-Gyu;Park, Ju-Young
    • The Journal of Korean Society for Radiation Therapy
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    • v.20 no.1
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    • pp.37-43
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    • 2008
  • Purpose: To evaluate dosimetry results of three different techniques for whole breast irradiation after conservative surgery of large pendulous breast patient. Materials and Methods: Planning computed tomography (CT) scans for three techniques were performed on a GE Hi-speed advantage CT scanner in the supine (SP), supine with breast supporting Device (SD) and prone position on a custom prone mattress (PP). Computed tomography images were acquired at 5 mm thickness. The clinical target volumes (CTV), ipsilateral lung and heart were delineated to evaluate the dose statistic, and all techniques were planned with the tangential photon beams (Pinnacle$^3$, Philips Medical System, USA). The prescribed dose was 50 Gy delivered in 25 fractions. To evaluate the dose coverage for CTV, we analysed percent volume of CTV receiving minimum of 95%, 100%, 105%, and 110% of prescription dose ($V_{95}$, $V_{100}$, $V_{105}$, and $V_{110}$) and minimal dose covering 95% ($D_{95}$) of CTV. The dosimetric comparison for heart and ipsilateral lung was analysed using the minimal dose covering 5% of each organs ($D_5$) and the volume that received >18 Gy for the heart and >20 Gy for the ipsilateral lung. Results: Target volume coverage ($V_{95}$ and $V_{100}$) was not significantly different for all technique. The V105 was lower for PP (1.2% vs. 4.4% for SP, 11.1% for SD). Minimal dose covering 95% ($D_{95}$) of target was 47.5 Gy, 47.7 Gy and 48 Gy for SP, SD and PP. The volume of ipsilateral lung received >20 Gy was 21.7%, 11.6% and 4.9% for SP, SD and PP. The volume of heart received >18 Gy was 17.0%, 16.1% and 9.8% for SP, SD and PP. Conclusion: Prone positioning of patient for large pendulous breast irradiation enables improving dose uniformity with minimal heart and lung doses.

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Bacteremia in Pediatric Cancer Patients : Causative Organisms and Antibiotic Sensitivities (소아 암 환자에서 발생한 균혈증 : 원인균과 항생제 감수성)

  • Kim, Yong-Han;Lee, Hyun-Dong;Hah, Jeong-Ok
    • Clinical and Experimental Pediatrics
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    • v.48 no.6
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    • pp.619-623
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    • 2005
  • Purpose : Bacteremia in immunocompromised pediatric cancer patients can lead to high morbidity and mortality, if not treated early and properly. The incidence and antibiotic sensitivities to common pathogens of bacteremia in pediatric cancer patients are liable to change, according to region and time. We investigated the causative organisms and antibiotic sensitivities of bacteremia in pediatric cancer patients to assess the adequacy of empiric antimicrobial therapy. Methods : From September 1995 to August 2003, we retrospectively evaluated 58 episodes in 39 pediatric cancer patients with bacteremia treated at the Pediatric Department of Yeungnam University Hospital. We investigated and analyzed the causative organisms and the antibiotic sensitivity test results by reviewing the records of the microbiologically proven positive blood culture results. Results : The incidence of bacteremia in pediatric cancer patients in this study was 5.7 percent (58 episodes out of 1,022 occasions of blood cultures). Gram-positive organisms were isolated more often than gram-negative organisms (63.8 percent vs 36.2 percent) in the following order : Staphylococcus epidermidis (37.9 percent), Staphylococcus aureus (17.3 percent), Escherichia coli (12 percent), Streptococcus (8.6 percent), Enterobacter (6.9 percent), Klesiella (6.9 percent), Serratia (3.5 percent), Acinetobacter (3.5 percent), Proteus (1.7 percent) and Morganella morganii (1.7 percent). In antibiotic sensitivity tests, only six of 37 isolates (16 percent) of gram positive bacteria were sensitive to penicillin and 15 of 37 isolates (40 percent) were sensitive to oxacillin. All except one Staphylococcus aureus were sensitive to vancomycin and all except one Staphylococcus epidermidis were sensitive to teicoplanin among 37 isolates of gram positive bacteria. In the case of gram negative bacteria, two of 21 isolates (10 percent) and four of 21 isolates (19 percent) were sensitive to cefotaxime and ceftazidime, respectively. Only six of 21 isolates (29 percent) were sensitive to aminoglycoside, but all 21 isolates (100 percent) were sensitive to imipenem. All seven isolates tested after the year 2000 were sensitive to meropenem. Conclusion : In conclusion, we should choose the proper antimicrobials in treating pediatric cancer patients with suspected bacteremia, reflecting the increasing episodes of gram positive bacteremia and polymicrobial resistance of gram positive and negative organisms.

Clinical Study on Fluvoxamine Combined with Oxycodone Prolonged-Release Tablets in Treating Patients with Moderate to Severe Cancer Pain

  • Xiao, Yang;Liu, Jun;Huang, Xin-En;Ca, Li-Hua;Ma, Yi-Min;Wei, Wei;Zhang, Rong-Xia;Huang, Xiao-Hong;Chang, Juan;Wu, Yi-Jia
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10445-10449
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    • 2015
  • Objective: To observe treatment effects and safety of fluvoxamine combined with oxycodone prolonged-release tablets in treating patients with moderate to severe cancer pain. Methods: Patients confirmed pathologically with cancer and complicated with moderate to severe pain, were divided into control and experimental groups. Oxycodone prolonged-release tablets, with or without fluvoxamine, were administrated to all study patients until pain relief. Degree of pain relief, dose of oxycodone prolonged-release tablets, side effects and quality of life were compared before and after treatment. Results: In total, 120 patients were recruited. No statistically significant difference was detected regarding age, gender, types of cancer, KPS between two groups of patients (P>0.05). Baseline pain score of patients with moderate pain in treatment and control group was $4.9{\pm}0.8$ and $5.1{\pm}0.8$, respectively; and decreased to $1.8{\pm}1.1$ and $1.2{\pm}1.1$ after treatment, respectively. Pain intensity was significantly reduced in the treatment group (P=0.028). Average daily consumption of oxycodone prolonged-release tablets was ($54.0{\pm}19.6$) mg and ($44.7{\pm}18.7$) mg respectively, which is lower in treatment grpup than in control group, but the difference was not statistically significant (P=0.065). Baseline pain score of patients with severe pain in treatment and control groups were $8.3{\pm}1.1$ and $8.3{\pm}1.1$, respectively; and pain intensity after treatment decreased to $2.9{\pm}1.0$ and $2.3{\pm}1.0$. Pain intensity was significantly reduced in the treatment group, with statistical significance (P=0.026). Average daily consumption of oxycodone prolonged-release tablets was ($132.0{\pm}42.2$) mg and ($110.7{\pm}33.9$) mg, respectively, which is lower in treatment group than in control group, and the difference was statistically significant (P=0.035). In terms of quality of life, patients in treatment group had better performance status, daily activity, mood, and sleep than that in control group (P < 0.05). Patients in two groups had similar side effects, eg., constipation, nausea/vomiting, lethargy, dizziness, itchy skin, dysuria, and ataxia. Lower incidence of nausea/vomiting, lethargy, was obtained from patients in treatment than in control group, while significant low constipation was observed in treatment than in control group (35.0% vs 49.2%, P=0.026). Conclusion: Fluvoxamine combined with oxycodone prolonged-release tablets could be more effective in treating patients with cancer pain, and could reduce the dosage of oxycodone prolonged-release tablets and thus be associated with lower side effects, and improved quality of life.

Comparison of Brain Activation Images Associated with Sexual Arousal Induced by Visual Stimulation and SP6 Acupuncture : fMRI at 3 Tesla (시각자극과 삼음교 자침으로 유발된 성적 흥분의 대뇌 활성화 영상의 비교 : 3 테슬라 기능적 자기공명영상법)

  • Choi, Nam-Gil;Han, Jae-Bok;Jang, Seong-Joo
    • Journal of radiological science and technology
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    • v.32 no.2
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    • pp.183-194
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    • 2009
  • Purpose : This study was performed not only to compare the brain activation regions associated with sexual arousal induced by visual stimulation and SP6 acupuncture, but also to evaluate its differential neuro-anatomical mechanism in healthy women using functional magnetic resonance imaging (fMRI) at 3 Tesla (T). Subjects and methods : A total of 21 healthy right-handed female volunteers (mean age 22 years, range 19 to 32) underwent fMRI on a 3T MR scanner. The stimulation paradigm for sexual arousal consisted of two alternating periods of rest and activation. It began with a 1-minute rest period, 3 minutes of stimulation with either of an erotic video film or SP6 acupuncture, followed by 1-minute rest. In addition, a comparative study on the brain activation patterns between an acupoint and a shampoint nearby GB37 was performed. The fMRI data were obtained from 20 slices parallel to the AC-PC line on an axial plane, giving a total of 2,000 images. The mean activation maps were constructed and analyzed by using the statistical parametric mapping (SPM99) software. Results : As comparison with the shampoint, the acupoint showed 5 times and 2 times higher activities in the neocortex and limbic system, respectively. Note that brain activation in response to stimulation with the shampoint was not observed in the regions including the HTHL in the diencephalon, GLO and AMYG in the basal ganglia, and SMG in the parietal lobe. In the comparative study of visual stimulation vs. SP6 acupuncture, the mean activation ratio of stimulus was not significantly different to each other in both the neocortex and the limbic system (p < 0.05). The mean activities induced by both stimuli were not significantly different in the neocortex, whereas the acupunctural stimulation showed higher activity in the limbic system (p < 0.05). Conclusions : This study compared the differential brain activation patterns and the neural mechanisms for sexual arousal, which were induced by visual stimulation and SP6 acupuncture by using 3T fMRI. These findings will be useful to understand the theory of traditional acupuncture and acupoint channel in scientific point of view.

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Complete Myocardial Revascularization Utilizing Parallel Secluential Anastomoses (Parallel sequential anastomosis를 이용한 관상동맥 우회수술의 성적)

  • 박계현;김관민
    • Journal of Chest Surgery
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    • v.30 no.7
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    • pp.647-655
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    • 1997
  • We evaluated the feasibility and safety of this method by reviewing the early outcome of the patients who underwent coronary artery bypass grafting(CABG) utili ing parallel sequential anastomoses with saphenous vein grafts, comparing with the outcome of the patients revascularized with grafts having only single distal anastomosis. During the one-year period of 1995, a total of 79 patients underwent isolated CABG, among whom 39 patients with sequential vein grafts(sequential group) and 40 patients without sequential grafts(non-sequential group). There was no difference between the two groups in terms of preoperative status, except in the extent of the coronary disease; 87.2% of the sequential group and 45.0% of the non-sequential group had left main andlor triple vessel involvement. 318 distal coronary anastomoses were done; 198 for the sequential group(5.1/patient) and 120 for the non-sequential group(3.0/patient). In the sequential group, the mean durations of cardiopulmonary bypass and aortic clamp per one distal anastomosis were 33.5 and 21.1 minutes, respectively. In the non-sequential group, these were 41.8 and 22.7 minutes. There were two operative deaths, both in the non-sequential group. There was no difference in the incidence of postoperative complications including myocardial infarction. During the follow-up period(2 to 15 months), 8 patients(3 in the sequential and 5 in the nonsequential group) complained of residual or recurrent angina. Comparison of preoperative and postoperative ."'Thallium myocardial perfusion scans in 30 patients showed improved or normal perfusion reserve in 83.3% of segments bypassed with sequential grafts and 82.5% of segments bypassed with non-sequential graft(5), These results show that, utilizing parallel sequential anastomoses with saphenous vein grafts, we could achieve satisfactory short-term clinical results in patients with extensive coronary stenoses. So, we conclude that this technique is a safe, technically feasible strategy for CABG, which can achieve the aim of complete myocardial revascularization with a limited length of graft.raft.

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The Clinical Characteristics of Endobronchial Anthracofibrotic Pigmentation (기관지 탄분 섬유화증의 임상양상)

  • Lim, Dongmee;Son, Jiwoong;Choi, Eugene
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.337-341
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    • 2006
  • Objectives : Endobronchial anthracofibrotic pigmentation, which presents as dark black or brown pigmentation mucosal changes of multiple bronchi combined with bronchial fibrosis and obstruction, is not a rare finding when performing diagnostic bronchoscopy for Koreans. This study was performed to define the clinical characteristics and to determine the association of these finding with the Korean life style and such other diseases as coal workers, pneumoconiosis or tuberculosis in the patients with anthracofibrotic pigmentation. Methods : This retrospective analysis was conducted on 70 (5.2%) patients with endobronchial anthracofibrotic pigmentation, among a total of 1340 patients who underwent bronchoscopy. The distinctive clinical features, the personal life style, the past medical history, the histology and microbiology, the radiologic finding and the natures of the bronchoscopic lesions were analyzed. Results : This mean age of the patients with anthracofibrotic pigmentation was $60.6{\pm}9.2$ year old and the male to female ratio was 1:1.7. The common respiratory symptoms of these patients were coughing and sputum (81%, 57/70), and this was followed in order by dyspnea and hemoptysisir. The symptoms were not related with smoking and an occupational history such as being a coal worker and so on. Pneumonia was most common finding on the radiologic studies. On bronchoscopy, the right middle lobe bronchus was most commonly involved. The most common associated disease was tuberculosis, and 40 cases (57.1%) were diagnosed by AFB staining, TB PCR, bronchoscopic guided tissue biopsy and a past history of tuberculosis. Other diseases related with anthracotic pigmentation were hypertension, diabetes, COPD, lung cancer, pneumoconiosis and asthma. Conclusion : These results suggest that endobronchial anthracofibrotic pigmentation was mostly related with pulmonary tuberculosis rather than with coal- related disease. Endobronchial anthracofibrotic pigmentation was more prevalent in older age females in Korea.