• Title/Summary/Keyword: laboratory method

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Effects of Embryo Sources and Culture Systems on the Membrane Permeability and Viability of Bovine Blastocysts Cryopreserved by GMP Vitrification (소 수정란의 생산체계가 세포막 투과력 및 GMP Vitrification 동결융해 후 생존성에 미치는 영향)

  • Kong, I.K.;Cho, S.G.
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.191-198
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    • 2001
  • The purpose of this study was to investigate the effects of embryo sources such as in vivo vs. in vitro produced blastocyst, and culture systems on the membrane permeability and viability of bovine blastocyst following GMP vitrification. To produce in vivo embryos, six cows were superovulated by administration of follicle stimulation hormone (FSH) and prostaglandin $F_{2{\alpha}}$(PG $F_{2{\alpha}}$). in vitro embryos were produced by two different culture systems, oviduct co-culture (OCS) and defined culture system (HECM-6; DCS). Ovaries were picked up at a local slaughterhouse and transported to laboratory in 3$0^{\circ}C$ saline within 2 h. Ovaries were washed with same saline three times and then placed in saline on warm plate adjusted at 3$0^{\circ}C$ during aspiration. The blastocysts produced were assigned for membrane permeability and viability following GMP vitrification. The membrane permeability of blastocysts was checked in 0.5 M sucrose solution on warm plate at 35$^{\circ}C$ for 0, 2, 5 and 7 min, respectively. Then the diameters (width and length) of embryo cytoplasms were measured by a eyepiece meter, and they were converted to their volume by 4/3 $\pi$ $r^3$. The blastocysts were cryopreserved by GMP vitrification method, where they were sequentially placed into vitrification solution before being loaded into GMP vessels and immersed into L$N_2$ within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for 5 min each, and then cultured in TCM 199 supplemented with 10% FCS for 24 or 48 h. The volume change of in vivo blastocyst at 0, 2, 5 and 7 min (100, 37.1, 34.3 and 31.6%) was significantly more shrunk than those of in vitro blastocysts derived from OCS (100, 59.8, 48.9, 47.9%) and DCS (100, 57.2, 47.3 and 46.9%) (P<0.05). The viability of post-thaw blastocyst derived from in vivo (93.6%) was also significantly different from those in OCS and DCS (81.9 and 83.6%; P<0.05). In the present culture system, the morphology of embryos produced in vitro was similar to that of in vivo embryos, but the quality in membrane permeability and post-thaw viability showed a big difference from their sources as in vivo or in vitro derived from OCS and DCS. The results indicated that the quality of in vivo embryos in membrane permeability and post-thaw viability was better than those of in vitro embryos derived from OCS or DCS.

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Effects of Deer Antler on the Regeneration of Peripheral Nerves; About Sprout Formation of Experimentally Transected Sciatic Nerves in Rat (말초신경의 재생에 대한 녹용의 효과; 랫드에서 실험적 절단 좌골신경의 Sprout 형성에 관해)

  • Chang, Byung-Joon;Cho, Ik-Hyun;Choi, Hye-Young;Won, Hui-Young;Park, Chang-Hyun;Bae, Chun-Sik;Choe, Nong-Hoon
    • Applied Microscopy
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    • v.32 no.1
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    • pp.67-80
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    • 2002
  • This study was carried out to investigate the effects of deer antler extract on the regeneration of peripheral nerves. Sprague-Dawley male rats weighing about 300 gm were fed deer antler extract for 1, 2, and 3 weeks per oral (1.5 ml/100 gm B.W.), respectively, once a day and transected both sides of sciatic nerve of each leg. After keeping for 6 hours, sciatic nerves taken from proximal part of transected region were treated with conventional transmission electron microscopical method and then observed with electron microscope. The results obtained were summarized as follows; 1. Sciatic nerves of normal control group were not showing any sprouts and electron dense axolemmal projections were frequently observed. 2. Sciatic nerves of saline treated groups were showing axonal sprouts at the nodes of Ranvier. The length of them was usually short, and numerous vesicles, vacuoles and organelles including neurofilament were contained. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 29 (29%) in 1 week treated group, 32 (32%) in 2 weeks treated group, and 30 (30%) in 3 weeks treated group, respectively. 3. Sciatic nerves of deer antler treated groups were showing axonal sprouts at the node of Ranvier as well. Although most of the sprouts were short, some sprouts of 2 weeks and 3 weeks treated groups were quite long. Sprouts usually contained numerous vesicles, vacuoles and cell organelles such as neurofilaments and mitochondria. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 38 (38%) in 1 week treated group, 46 (46%) in 2 weeks treated group, and 48 (48%) in 3 weeks treated group respectively. The results described above explain pretreatment of deer antler extract improves the sprout formation of transected sciatic nerves, and then it suggests deer antler may be effective for the regeneration of peripheral nerves.

Organic Acidopathies as Etiologic Diseases of Developmental Delay in Korean Childhood and Adolescent Age Group (한국인 소아청소년기 발달지연의 원인질환으로서의 유기산대사이상질환)

  • Lee, Jong Yoon;Lee, Ye Seung;Choi, Joong Wan;Bae, Eun Joo;Park, Won Il;Oh, Phil Soo;Lee, Hong Jin
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.12 no.2
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    • pp.85-93
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    • 2012
  • Purpose: Developmental delay is caused by very diverse etiologic diseases. Most chronic disorders has some influence on development. Chronic or acute disorders of CNS are main etiologic diseases of developmental delay. Up to now, over 60 diseases are included in organic acidopathies and most of them causes acute or chronic recurrent CNS damage and developmental delay. We have done this study to find out the importance of organic acidopathies causing developmental delay in Korean childhood and adolescent patients. Method: Retrograde analysis for 738 patients with developmental delay whose clinical informations are available and have done urine organic acid analysis for 5 years period, between Jan. 1st 2007 to Dec. 31th 2011. Statistical analysis was done with Student's t test using SPSS. Result: Out of 738 patients, 340 patients (46.1%) showed abnormalities on urine organic acid analysis. The most frequent disease was mitochondrial respiratory chain disorders (MRCD) (253, 34.3%), followed by ketolytic defects(39, 5.3%), 3-hydroxyisobutyric aciduria (26, 3.5%), glutaric aciduria type II (8, 1.1%), pyruvate dehydrogenase deficiency (3, 0.4%), 3-methylglutaric aciduria (2, 0.3%), glutaric aciduria type I (2, 0.3%), ethylmalonic aciduria (1, 0.15%), methylmalonic aciduria (1, 0.15%), HMG-CoA lyase deficiency (1, 0.15%), 3-methylcrotonylglycinuria (1, 0.15%), fatty acid oxidation disorders(1, 0.15%) and FAOD (1, 0.15%). Conclusion: Mitochondrial disorders are most frequent etiologic disease on all age group, followed by ketolytic defects and various organic acidopathies. The number and diversities of organic acidopathies emphasize meticulous evaluation of basic routine laboratory examinations and organic acid analysis with initial sample on every developmental patient.

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Assessment of Nitrogen Impaction on Watershed by Rice Cultivation (벼농사에서 질소유출이 수질에 미치는 영향평가)

  • Roh, Kee-An;Kim, Min-Kyeong;Lee, Byeong-Mo;Lee, Nam-Jong;Seo, Myung-Chul;Koh, Mun-Hwan
    • Korean Journal of Environmental Agriculture
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    • v.24 no.3
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    • pp.270-279
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    • 2005
  • It is important to understand and evaluate the environmental impacts of rice cultivation for developing environmentally-friendly agriculture because rice is main crop in Korea and rice cultivation have both functions of water pollution and purification with environmental and cultivation conditions. This paper presents the evaluation of nitrogen impact by rice cultivation on water system. A simple protocol was proposed to assess the potential amount of nitrogen outflow from paddy field and most of parameters affect on the nitrogen outflow from paddy field such as the amount of fertilizer application, water balance, the quality and quantity of irrigation water, soil properties, nitrogen turnover in the soil and cultivation method were considered. To develop the protocol, coefficients for parameters affected nitrogen turnover and outflow were gotten and summarized by comparison and analysis of all possible references related, and by additional experiments at field and laboratory. And potential amount of nitrogen input and output by water in paddy field were estimated with the protocol at the conditions of the nitrogen contents of irrigation water, amount of fertilizer application, and irrigation methods. Where irrigation water was clean, below 1.0 mg $L^{-1}$ of nitrogen concentration, rice cultivation polluted nearby watershed. At the conditions of 2.0 mg $L^{-1}$ of nitrogen concentration, 110 kg $ha^{-1}$ of nitrogen fertilizer application and flooding irrigation, rice cultivation had water pollution function, but it had water purification function with intermittent irrigation. At the conditions of 3.0 mg $L^{-1}$ of nitrogen concentration and 110 kg $ha^{-1}$ of nitrogen fertilizer application, rice cultivation had water purification function, but that had water pollution function with 120 kg $ha^{-1}$ of nitrogen application. Where irrigation water was polluted over 6.0 mg $L^{-1}$ of nitrogen, it was evaluated that rice cultivation had water purifying effect, even though the amount of nitrogen application was 120 kg $ha^{-1}$.

Clinical usefulness of Clinical Pulmonary Infection Scare of ICU Patients with Sputum Culture positive for Multi-drug resistant Acinetobacter baumannii (객담에서 Multi-drug Resistant Acinetobacter baumannii가 동정된 중환자실 환자에서 Clinical Pulmonary Infection Score의 임상적 유용성)

  • Lee, Ji Hyun;Cheon, Seok Cheol;Jung, Sun Hye;Phyun, Lae Hyun;Jang, Moon Zu;Lee, Stephen Yonggu;Hong, Sung Kwan;Hong, Seong Geun;Hong, Sang Bum
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.6
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    • pp.579-588
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    • 2003
  • Background : The hospital-acquired pneumonia is the most common nosocomial infection. Recently, the Acinetobacter baummannii infections are rapidly increasing, especially the frequency of Multi-drug resistant A. baumannii. Therefore we assessed clinical features and prognosis of patients in the ICU with Multi-drug resistant A. baumannii from the sputum culture using the Clinical Pulmonary Infection Score(CPIS). Method : The medical records of 43 patients with Multi-drug resistant A. baumannii from sputum culture who were suspected had clinically pneumonia and admitted to the ICU from January 2000 to July 2002 were retrospectively analyzed. Results : 19 patients were CPIS greater than 6 and 24 patients were CPIS less than or equal to 6. Mean age for the former was $71{\pm}11$ years old, and the latter was $61{\pm}19$ years old. The mean APACHE II score on admission and on sputum study was not different between two groups($17.4{\pm}5.7$ vs $18.5{\pm}6.1$, p=0.553, $20{\pm}6$ vs $17{\pm}8$, p=0.078). But the mortality rate was 73.7% for the former, and 16.7% for the latter(p<0.001). Conclusion : In ICU patients who had clinically suspected pneumonia with sputum culture positive for Multi-drug resistant A. baumannii, the mortality was significantly higher if CPIS was greater than 6.

Measurement of Nitric Oxide in the Differential Diagnosis of Lymphocytic Pleural Effusion (림프구성 흉막염의 감별 진단에서 NO(nitric oxide)의 측정)

  • Kim, Tae-Hyung;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.4
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    • pp.361-367
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    • 2005
  • Background : Differential diagnosis of lymphocytic pleural effusion is difficult even with many laboratory findings. Nitric oxide(NO) level is higher in the sputum or exhaled breath of patients with active pulmonary tuberculosis than in those without tuberculosis. In addition, there are some reports about the increased level of NO metabolites in body fluids of cancer patients. However, there is no data on the NO levels in the pleural fluid of patients with tuberculous pleurisy. Method : The serum and pleural fluid NO in the patients with acute lymphocytic pleural effusion were analyzed. Results : Of total 27 patients, there were 14 males and average age of patients was 48 years. The final diagnosis was tuberculous pleurisy in 17 cases and malignant pleural effusion in 10. The pleural fluid NO level was $540.1{\pm}116.4{\mu}mol$ in the tuberculous pleurisy patients and $383.7{\pm}71.0{\mu}mol$ in the malignant pleural effusion patients. The serum NO level was $624.7{\pm}142.0{\mu}mol$ in tuberculous pleurisy patients and $394.4{\pm}90.4{\mu}mol$ in malignant pleural effusion patients. There was no significant difference in the serum and pleural fluid NO level between the two groups. The NO level in the pleural fluid showed a significant correlations with the pleural fluid neutrophil count, the pleural fluid/serum protein ratio, and pleural fluid/serum albumin ratio (p<0.05 in each). The protein concentration, leukocyte and lymphocyte count in the pleural fluid were significantly higher in the tuberculous pleurisy patients than the malignant pleural effusion patients (p<0.05 in each). Conclusion : NO is not a suitable marker for a differential diagnosis of lymphocytic pleural effusion. However, the NO level in the pleural fluid might be associated with the neutrophil recruitment and protein leakage in the pleural space.

Repeatability and Reproducibility in Effective Porosity Measurements of Rock Samples (암석시험편 유효공극률 측정의 반복성과 재현성)

  • Lee, Tae Jong;Lee, Sang Kyu
    • Geophysics and Geophysical Exploration
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    • v.15 no.4
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    • pp.209-218
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    • 2012
  • Repeatability and reproducibility in solid weight and effective porosity measurements have been discussed using 8 core samples with different diameters, lengths, rock types, and effective porosities. Further, the effect of temperature on the effective porosity measurement has been discussed as well. Effective porosity of each sample has been measured 7 times with vacuum saturation method with vacuum pressure of 1 torr and vacuum time of 80 minutes. Firstly, effective porosity of each sample is measured one by one, so that it can provide a reference value. Then for reproducibility check, effective porosity measurements with vacuum saturation of 2, 4, and 8 samples simultaneously have been performed. And finally, repeated measurements for 3 times for each sample are made for repeatability check. Average deviation from the reference set in solid weight showed 0.00 $g/cm^3$, which means perfect repeatability and reproducibility. For effective porosity, average deviations are less than 0.07% and 0.05% in repeatability and reproducibility test sets, respectively, which are in good agreement too. Most of porosities measured in reproducibility test lies within the deviation range in repeatability test sets. Thus, simultaneous vacuum saturation of several samples has little impact on the effective porosity measurement when high vacuum pressure of 1 torr is used. Air temperature can cause errors on submerged weight read and even effective porosity, because it is closely related to the temperature, density, and buoyancy of water. Consequently, for accurate measurement of effective porosity in a laboratory, efforts for maintaining air or water temperature constant during the experiment, or a temperature correction from other information are needed.

Molecular Characterization and Phylogenetic Analysis of Season Influenza Virus Isolated in Busan during the 2006-2008 Seasons (부산지역에서 유행한 계절인플루엔자바이러스의 유전자 특성 및 계통분석('06-'08 절기))

  • Park, Yon-Koung;Kim, Nam-Ho;Choi, Seung-Hwa;Lee, Mi-Oak;Min, Sang-Kee;Kim, Seong-Joon;Cho, Kyung-Soon;Na, Young-Nan
    • Journal of Life Science
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    • v.20 no.3
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    • pp.365-373
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    • 2010
  • To monitor newly emerged influenza virus variants and to investigate the prevalence pattern, our laboratory performed isolation of the viruses from surveillance sentinel hospitals. In the present study, we analysed influenza A/H1N1, A/H3N2, B viruses isolated in Busan during the 2006/07 and 2007/08 seasons by sequence analysis of the hemagglutinin (HA1 subunit) and neuraminidase (NA) genes. The isolates studied here were selected by the stratified random sample method from a total of 277 isolates, in which 15 were A/H1N1, 16 were A/H3N2 and 29 were B. Based on the phylogenetic tree, the HA1 gene showed that A/H1N1 isolates had a 96.7% to 97.7% homology with the A/Brisbane/59/2007, A/H3N2 isolates had a 98.4% to 99.7% homology with the A/Brisbane/10/2007, and B isolates had a 96.5% to 99.7% homology with the B/Florida/4/2006(Yamagata lineage), which are all the vaccine strains for the Northern Hemisphere in 2008~2009 season. In the case of the NA gene, A/H1N1 isolates had 97.8% to 98.5% homologies, A/H3N2 isolates had 98.9% to 99.4% homologies, and B isolates had 98.9% to 100% homologies with each vaccine strain in the 2008~2009 season, respectively. Characterization of the hemagglutinin gene revealed that amino acids at the receptor-binding site and N-linked glycosylation site were highly conserved. These results provide useful information for the control of influenza viruses in Busan and for a better understanding of vaccine strain selection.

Prevalence and Infection Status of Salmonella in 25 Conventional Swine Farms in Korea (국내 25개 양돈장의 살모넬라 유병율 및 감염유형)

  • Park, Choi-Kyu;Kim, Hee-Jung;Kim, Jin-Hyun;Cho, Jae-Keun;Kim, Young-Hwa;Jung, Yoon-Soo;Bae, Chae-Wun;Park, Jun-Cheol;Kim, In-Cheul;Kim, Ki-Seuk
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1267-1272
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    • 2013
  • The aim of this study was to determine the prevalence and infection status of Salmonella species (spp.) in 25 conventional pig farms by traditional fecal culture and serological methods to develop a Salmonella control program for Korean pig farms. The individual seroprevalence of Salmonella spp. in pigs reared in the 25 pig farms was 83.1% in sows and 6.4-32% in different aged pig groups, with the total seroprevalence 28.4% (141/848). The seroprevalence of the tested pigs increased in accordance with the decrease in maternal antibody and the rearing period on these farms. Of note, all the 25 pig farms contained at least two or more anti-Salmonella antibody-positive sows. In the fecal cultures Salmonella spp. were isolated only in three (12.0%, 3/25) of 16 serologically Salmonella-suspected farms (64.0%, 16/25), showing the limitation of the fecal culture method and the need for serum assays to understand the exact status of Salmonella infection in swine herds, which likely contain subclinically infected pigs or carriers. The results highlight the need to establish a supply system of Salmonella-free gilts for the promotion of a national Salmonella control program on swine farms in Korea. Further studies will be needed to develop an effective monitoring system for the implementation of a national Salmonella control program.

Efficacy of Antibacteriocidal Yeast That Producing Bacteriocin OR-7 in Chicken (박테리오신 OR-7을 생산하는 항균 효모의 양계에서의 사양시험 효과)

  • Cho, Dong-In;Kang, Sang-Mo;Lee, Jae-Hwa;Lee, Sang-Hyeon;Kim, Nam-Young;Kim, Ki-Seuk
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.391-398
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    • 2010
  • This study was conducted to investigate the effects of feeding probiotics(gene modified yeast) on the egg quality in laying hens. The laboratory method of this study is as follow: In hens, 300, 36 weeks old ISA brown commercial layer, were employed in 13weeks feeding trial with a 7 days adjustment period. Dietary treatments are 1) control (basal diet) 2) Y0.3 (basal diet+0.3% probiotics), 3) Y0.5% (basal diet+0.5% probiotics), 4) PY0.3% (basal diet+0.3% plasmid modified probiotics), 5) PY0.5% (basal diet+0.5% plasmid modified probiotics). For overall period, hen-day egg production, egg weight (p<0.05) by dietary probiotic supplementation were recorded. Eggs were collected and weighed every day. Egg production number and egg production rate, egg weight, failure egg ratio and trouble egg ratio were recorded for 13weeks days. Diets PY0.3 and PY0.5 improved Egg production rate, egg production number and egg weight, compared to control diet (linear effect. p<0.05). And, diets PY0.3 and PY0.5 improved Egg production rate, egg production number, compared to Y0.3 and Y0.5 (linear effect. p<0.05). But, in Inferior egg and cracked egg, diets PY0.3 and PY0.5 did not tended to increase by dietary probiotic supplementation compared to control diet and Y group (0.3, 0.5) (linear effect. p<0.05). The productivity enhance on the egg quality in laying hens is considered from the effects of feeding probiotics(gene modified yeast).