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Histopathological and Serological Observations on Experimental Anisakiasis of Rabbits (실험적 토끼 아니사키스증에서의 조직학적 병변과 혈청 항체가의 경시적 변동 양상)

  • 홍성태;이손형
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.168-180
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    • 1987
  • This study was performed to observe histopathological changes and serological reactions in chronic anisakiasis of rabbits. Each rabbit was infected per os with 30 larvae of Anisakis type I. Their sera were collected chronologically and the rabbits were killed for histopathological examination, 3, 13, 20, 30, 60, 90 and 150 days after the infection. The results were summarized as below. 1. Most of the larvae were recovered from the stomach, but a few from the omentum, intestine, mesentery and abdominal wall. The recovery rates and distribution of worms by organ were not differed by duration of infection. 2. Histologically the lesion was abscess type on 13 days, i.e., the dead worms were surrounded by fibrinous exudate, histiocytes and thick zone of numerous inflammatory cells. After 30 days, histiocytes were found to invade the worms and the lesion was changing into abscessgranulomatous type. Also a calcified worm was found on the 30th day. After then the worms were observed to be dissolved slowly until 90 days. On 150 day, only one calcified worm was observed. 3. The levels of serum IgG antibody by ELISA reached their maximum 30 days after the infection. After then, it decreased slowly until 150 days after the infection. Above serological and histopathological findings indicated that antigenic stimulation from degenerating Anisakis larvae was the greatest during the first 30 days after infection. This period was corresponding with the beginning of worm resolution or calcification. Serologic test by ELISA would be a valuable tool for confirming chronic anisakiasis.

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Surface maker and gene expression of human adipose stromal cells growing under human serum. (인체혈청 하에서 배양한 인체지방기질줄기세포의 표면항원 및 유전자 발현)

  • Jun, Eun-Sook;Cho, Hyun-Hwa;Joo, Hye-Joon;Kim, Hoe-Kyu;Bae, Yong-Chan;Jung, Jin-Sup
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.678-686
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    • 2007
  • Human mesenchymal stem cells(hMSC), that have been reported to be present in bone marrow, adipose tissues, dermis, muscles and peripheral blood, have the potential to differentiate along different lineages including those forming bone, cartilage, fat, muscle and neuron. Therefore, hMSC are attractive candidates for cell and gene therapy. The optimal conditions for hMSC expansion require medium supplemented with fetal bovine serum(FBS). Some forms of cell therapy will involve multiple doses, raising a concern over immunological reactions caused by medium-derived FBS proteins. Previously, we have shown that hADSC can be cultured in human serum(HS) during their isolation and expansion, and that they maintain their proliferative capacity and ability for multilineage differentiation and promote engraftment of peripheral blood-derived CD34 cells mobilized from bone marrow in NOD/SCID mice. In this study we determined whether hADSC grown in HS maintain surface markers expression similar with cells grown in FBS during culture expansion and compared gene expression profile by Affymetrix microarray. Flow cytometry analysis showed that HLA-DR, CD117, CD29 and CD44 expression in HS-cultured hADSC during culture expansion were similar with that in FBS-cultured cells. However, the gene expression profile in HS-cultured hADSC was significantly different from that in FBS-cultured cells. Therefore, these data indicated that HS-cultured hADSC should be used in vivo animal study of hADSC transplantation for direct extrapolation of preclinical data into clinical application.

Removal of Nitrogenous Compounds by Immobilized Mixed Microorganisms Including Photosynthetic Bacteria (광합성 세균을 포함한 고정화 복합미생물에 의한 질소성분 제거)

  • Cho, Kyoung Sook;Kim, Jeong Bo;Jeong, Soo Kyoung;Jeong, Hae Yoon;Cho, Jeong Sub;Kim, Joong Kyun
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.91-97
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    • 2006
  • For efficient removal of nitrogenous compounds produced in recirculating aquaculture system, the N removal characteristics of immobilized mixed microorganisms were investigated at various mixing ratios of photosynthetic bacteria (PSB) immobilized in PVA beads or CTA cubes and ammonium utilizing bacteria (AUB) immobilized in PVA beads. On the optimal medium of AUB, the maxium gas production rate was obtained at the mixing ratio of 10:40 (PSB:AUB), and the gas production rate increased as the portion of AUB beads in the mixed beads increased. When the mixing ratios of PSB:AUB beads were 50:0, 40:10, 25:25 and 10:40, the final pHs were measured to be 6.29, 6.01, 5.69 and 5.13, respectively. On the optimal medium of PSB, however, the volume and the rate of gas production decreased remarkably as the portion of AUB beads in the mixed beads increased. The final pH was measured to be approximately 6.5, regardless of the mixing ratio. In the reactions by the mixed culture of PSB cubes and AUB beads, all results showed the same tendency of those by the mixed culture of PSB and AUB beads, but the volume and the rate of gas production decreased remarkably, even with 0.2ml of gas production in control. From all the results, the use of mixed PSB and AUB beads at the ratio of 10:40 seems to be efficient to remove nitrogenous compounds in wastewater from recirculating aquaculture system.

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Preparation of $^{99m}Tc-HYNIC-PEG-liposomes$ for Imaging of the Focal Sites of Infection (농양 진단을 위한 $^{99m}Tc-HYNIC-PEG-liposomes$의 제조)

  • Hong, Jun-Pyo;Awh, Ok-Doo;Kim, Hyun-Suk;Lee, Eun-Sook;Lee, Tae-Sup;Choi, Tae-Hyun;Choi, Chang-Woon;Lim, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.6
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    • pp.333-343
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    • 2002
  • Purpose: A new linker, hydrazino nicotinamide (HYNIC), was recently introduced for labelling of liposome with $^{99m}Tc$. In this study we synthesized HYNIC derivatized PEG (polyethylene glycol)-liposomes radiolabeled with $^{99m}Tc$. Materials and Methods: In order to synthesize HYNIC-DSPE (distearoyl phosphatidyl ethanolamine) which is a crucial component for $^{99m}Tc$ chelation, first of all succinimidyl 6-BOC-hydrazinopyridine-3-carboxylic acid was synthesized from 6-chloronicotinic acid by three sequential reactions. A DSPE derivative of succinimidyl 6-BOC-hydrazinopyridine-3-carboxylic acid was transformed into HYNIC-DSPE by HCI/dioxane. HYNIC-PEG-liposomes were prepared by hydration of the dried lipid mixture of EPC (egg phosphatidyl choline): PEG-DSPE : HYNIC-DSPE:cholesterol (1.85:0.15:0.07:1, molar ratio). The HYNIC-PEG-liposomes were labeled with $^{99m}Tc$ in the presence of $SnCl_2{\cdot}2H_2O$ (a reducing agent) and tricine (a coligand). To investigate the level of in vivo transchelation of $^{99m}Tc$ in the liposomes, the $^{99m}Tc$-HYNiC-PES-liposomes were incubated with a molar excess of DTPA, cysteine or glutathione solutions at $37^{\circ}C$ for 1 hour. The radiolabeled liposomes were also incubated in the presence of human serum at $37^{\circ}C$ for 24 hours. Results: 6-BOC-hydrazinopyridine-3-carboxylic acid was synthesized with 77.3% overall yield. The HYNIC concentration in the PEG-coated liposome dispersion was 1.08 mM. In condition of considering the measured liposomal size of 106 nm, the phospholipid concentration of $77.5\;{\mu}mol/m{\ell}$ and the liposomal particle number of $5.2{\times}10^{14}$ liposomes/ml, it is corresponded to approximate 1,250 nicotinyl hydrazine group per liposome in HYNIC-PEG-liposome. The removal of free $^{99m}Tc$ was not necessary because the labeling efficiency were above 99%. The radiolabeled liposomes maintained 98%, 96% and 99%, respectively, of radioactivity after incubation with transchelators. The radiolabeled liposomes possessed above 90% of the radioactivity in serum. Conclusion: These results suggest that the HYNIC can be synthesized easily and applied in labelling of PEG-liposomes with $^{99m}Tc$.

Estimation of Oxygen Consumption Rate and Organic Carbon Oxidation Rate at the Sediment/Water Interface of Coastal Sediments in the South Sea of Korea using an Oxygen Microsensor (산소 미세전극을 이용한 남해연안 퇴적물/해수 계면에서 산소소모율 및 유기탄소 산화율 추정)

  • Lee, Jae-Seon;Kim, Kee-Hyun;Yu, Jun;Jung, Rae-Hong;Ko, Tae-Seung
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.8 no.4
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    • pp.392-400
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    • 2003
  • We used an oxygen microelectrode to measure the vertical profiles of oxygen concentration in sediments located near point sources of organic matter. The measurements were carried out between 13th and 17th May, 2003, in semi-closed bay and coastal sediments in the central part of the South Sea. The measured oxygen penetration depths were extremely shallow and ranged from 1.30 to 3.80 mm. This suggested that the oxidation and reduction reactions in the early diagenesis should be studied at the mm depth scale. In order to estimate the oxygen consumption rate, we applied the one-dimension diffusion-reaction model to vertical profiles of oxygen near the sediment/water interface. Oxygen consumption rates were estimated to be between 10.8 and 27.6 mmol O$_2$ m$\^$-2/ day$\^$-1/(average: 19.1 mmol O$_2$ m$\^$-2/ day$\^$-1/). These rates showed a positive correlation with the organic carbon of the sediments. The corresponding benthic organic carbon oxidation rates calculated using an modified Redfield ratio (170/110) at the sediment/water interface were in the range of 89.5-228.1 mg C m$\^$-2/ day$\^$-1/(average: 158.0 mg C m$\^$-2/ day$\^$-1/). We suggest that these results are maximum values at the presents situation in the bay because the sampling sites were located near point sources of organic materials. This study will need to be carried out at many coastal sites and throughout the seasons to allow an understanding of the mechanisms of eutrophication e.g. the spatial distribution of oxygen consumption within the oxic zone and hypoxic conditions in the coastal sea.

Studies on the Volatile Flavor Compounds of Sesame Oils with Roasting Temperature (볶음온도에 따른 참기름의 휘발성향기성분 변화)

  • Kim, Hyeon-Wee;Park, Ki-Moon;Choi, Chun-Un
    • Korean Journal of Food Science and Technology
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    • v.32 no.2
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    • pp.238-245
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    • 2000
  • This study was investigated to compare the changes of flavors in sesame oil with roasting temperature $(110^{\circ}C{\sim}230^{\circ}C)$. In the results of analyzing the volatile flavor compounds of sesame oil with GC and GC/MS, 26 pyrazines, 11 pyridines, 9 thiazoles, 6 furans, 8 pyrroles, 5 phenols, 8 aldehydes, 8 hydrocarbons, 7 alcohols, 2 indoles, 3 ketones, 10 acids, 4 nitriles, 7 esters, and 5 others were isolated, identified, and quantified. The total amount of flavor compounds was increased with roasting temperature. Detected flavors could be devided into top(peak No. $1{\sim}91$), middle$(92{\sim}197)$ and last note$(198{\sim}224)$ by rentention time. The top notes(initial content 19.87 ppm) which contain pyrazines and provide representative roasted flavors were increased significantly with roasting temperature. Initial content of middle note(17.72 ppm) was increased to 36.71 ppm at $170^{\circ}C$, to 95.61 ppm at $220^{\circ}C$, and to 138.62 ppm at $230^{\circ}C$. Last note was almost unchanged up to $170^{\circ}C$ and increased at $190^{\circ}C$, whereas it indicated a tendency to decrease at $230^{\circ}C$. Pyrazines such as methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, trimethylpyrazine, 2-ethyl-3,5-dimethylpyrazine which indicate the major components among volatile flavors were increased slightly up to $150^{\circ}C$ and revealed the higher increase than any other components above $170^{\circ}C$. This tendency was also similar to pyridines, thiazoles, and furans. Most of these compounds are assumed to be developed by thermochemical reactions of sesame components by roasting above $170^{\circ}C$. It seemed that a lot of increase in phenols above $210^{\circ}C$ resulted from the production of guaiacol. Acids were almost unchanged up to $190^{\circ}C$, increased at $210^{\circ}C$, and then decreased above $220^{\circ}C$. It seemed to be resulted from pyrolysis of free fatty acids formed from thermal oxidation of oil.

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Studies on Witches' Broom of Ligustrum ovalifolium Hasskarl Caused by Mycoplasma-like Organism (MLO) (Mycoplasma성(性) 왕쥐똥나무 빗자루병(病)에 관(關)한 연구(硏究))

  • Chai, Jyung-Ki;Kim, Young-Ho
    • Journal of Korean Society of Forest Science
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    • v.78 no.2
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    • pp.103-118
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    • 1989
  • The occurrence of witches' broom in Ligustrum ovalifolium was first noticed in Korea by author in 1984. The present study was carried out with particular emphasis on the symptomatology, etiology, transmission of the disease and antibiotic treatments. The infected tissue was observed by the fluorescence and electron microscopy and its biochemical characteristics were compared with healthy one by electrophoresis. The results are summarized as follows : 1. symptoms of the infected trees were characterized by the dwarfing of the organs, yellowing and brooming of the foliage. 2. The observation by the trans electron microscopy on the witches' broom of L. ovalifolium revealed the occurrence of numerous mycoplasma-like organisms(MLOs) in the phloem tissue cells of the midribs of infected leaves. 3. The MLOs were surrounded by a single unit membrane, and they appeared to be multiplied by binary fission. 4. The presence of crystals unidentified in the phloem parenchyma cells was noticed by electron rnicroscopy, 5. The disease was able to be transmitted by budding, crown, and greenwood graftings to L. ovalifolium, L. obtusifolium, L, japonicum and also transmitted, even when the stocks and scions were not completely grafted. 6. Insect transmission on L. ovalifolium and L, obtzrsifolium was carried by Hishimonus sellatus. 7. The infected roots dipped in the 1,000 ppm of teracyclin solution was only temporarily effective in controlling the disease. 8. Infected plant with MLOs showed specific fluorescent reactions in phloems with DAPI stain. 9. The protein and peroxidase separated by electrophoresis showed strikingly distinctive difference between the healthy and diseased leaves.

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Evaluation of Peptide Nucleic Acid Probe-Based Fluorescence In Situ Hybridization for the Detection of Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria in Clinical Respiratory Specimens (임상 객담검체에서 Peptide Nucleic Acid Probe를 이용한 결핵과 비결핵 항산균의 구분)

  • Lee, Seung Hee;Kim, Shine Young;Kim, Hyung Hoi;Lee, Eun Yup;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.18 no.2
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    • pp.37-43
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    • 2015
  • Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.

Development and Preliminary Evaluation of a Leukocyte Removal Aptamer Filter (압타머를 이용한 백혈구제거필터의 개발 및 예비평가)

  • Lee, Yangwon;Jung, Eun-Suk;Choi, Kyoung Young;Kim, Myung Han;Kwon, So Yong;Cho, Nam Sun;Kim, Jin Sook;Park, Han Jeong;Han, Byoung Don;Yoon, Soo-Young
    • The Korean Journal of Blood Transfusion
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    • v.23 no.2
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    • pp.107-114
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    • 2012
  • Background: Leukocyte reduction filters are widely used to prevent transfusion reactions caused by leukocytes in blood components. Commercial filters are not sufficient for removal of leukocytes for prevention of transfusion associated graft-versus-host disease; therefore, irradiation of blood components was performed using expensive equipment. Techniques using an aptamer substituted for antibody have been developed and are available in clinical areas. The purpose of this study is to develop the aptamer filter system and to evaluate its efficiency and the possibility of its clinical application. Methods: Aptamers targeted to CD45 were selected by the Postech Aptamer Initiative. The aptamer filter in which aptamers attached to beads were bound to leukocytes and removed by magnetic field was developed. Filtration of 14 units of leukoreduction-red blood provided by Korean Red Cross Blood Services was performed using aptamer filters. Leukocyte removal rate and red cell recovery rate were evaluated and bacterial culture was performed. Results: After filtration using the aptamer filters, 45.6% of leukocytes were additionally removed and the red cell recovery rate was 92.8%. No growth in the bacterial culture was observed. Conclusion: In order to apply the cell depletion technique utilizing an aptamer to blood filter system, we developed and evaluated the aptamer filter system. Through improvement of the binding efficiency of the aptamer and the filtering process, and application of the various aptamers for other different cells, we suggest that this technique can be applied in the clinical area, such as a substitution for the irradiation process for TAGVHD prevention.

Intravitreal Anti-vascular Endothelial Growth Factor Injections to Treat Neovascular Age-related Macular Degeneration: Long-term Treatment Outcomes (삼출 나이관련황반변성에 대한 항혈관내피성장인자 유리체내주사 치료의 장기 임상 결과)

  • Park, Yu Jeong;Son, Gi Sung;Kim, Yoon Jeon;Kim, June-Gone;Yoon, Young Hee;Lee, Joo Yong
    • Journal of The Korean Ophthalmological Society
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    • v.59 no.12
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    • pp.1142-1151
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    • 2018
  • Purpose: We assessed the visual and anatomical outcomes, and the safety profile of long-term intravitreal anti-vascular endothelial growth factor (VEGF) injections (aflibercept, ranibizumab, and bevacizumab) given to treat neovascular age-related macular degeneration (NAMD). Methods: We analyzed medical records collected over 7 years of treatment-naive NAMD patients who received outpatient clinic-based intravitreal anti-VEGF injections. All were treated employing either "treat-and-extend" or "as needed" protocols at the discretion of the retinal specialist. The number of injections, adverse events associated with injection, and measures of visual acuity (VA), central foveal thickness (CFT), and intraocular pressure (IOP) were recorded. Results: Overall, we assessed 196 eyes of 196 patients (average age $68.6{\pm}9.6years$; 77 females). Patients received an average of $17.3{\pm}13.5$ injections over $78.0{\pm}16.5months$ of clinical follow-up. The initial mean VA (logMAR) was $0.75{\pm}0.58$ and the CFT was $349.7{\pm}152.6{\mu}m$. Both parameters exhibited maximal improvements at the 6-month visit (p < 0.05). However, the clinical outcomes worsened over the 7-year clinical course; the best-corrected visual acuity (BCVA) was $0.91{\pm}0.78$ and the CFT was $284.5{\pm}105.8{\mu}m$ at 7 years. The BCVA at 7 years was significantly correlated with the initial BCVA. IOP-related events increased 11-fold and anterior chamber reactions increased 3-fold over the years, but no significant complications such as endophthalmitis were recorded. Conclusions: The use of intravitreal anti-VEGF agents was associated with initial visual improvements over 6 months but did not prevent the worsening of NAMD over 5 years. The BCVA at the initial visit was a strong predictor of the final BCVA. A more intensive injection schedule might improve long-term outcomes.