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Detection of Human Taurine Transporter and Production of Monoclonal Antibody

  • An, Hye-Suk;Han, Hee-Chang;Lee, Sun-Min;Park, Taesun;Park, Kun-Koo;Kim, Ha-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.102-102
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    • 2001
  • Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

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Analysis of Growth and Hematologic Characteristics of Cloned Puppies (체세포 복제 자견의 성장 및 혈액학적 특성 분석)

  • Kim, Dong-Hoon;Choi, Mi-Kyung;No, Jin-Gu;Park, Jong-Ju;Yeom, Dong-Hyeon;Kim, Hyun-Min;Choi, Bong-Hwan;Kim, Dong-Kyo;Park, Jin-Ki;Yoo, Jae Gyu
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.229-235
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    • 2013
  • The objective of this study was to monitor health conditions of four genetically identical somatic cells cloned Labrador retriever puppies by estimation of body weight and analysis of hematologic and serologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and four live puppies were delivered by Caesarean section. The cloned Labrador retrievers were genetically identical to the nuclear donor dog. The body weight of clone-1, -2, -3, and -4 was increased from 0.66, 0.40, 0.39, and 0.37 kg at birth to 6.2, 6.6, 6.2, and 6.0 kg at 8 weeks of age, respectively. Although clone-4 had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned puppies were not different when compared to reference range. In serological analysis, Glucose, ALP and inorganic phosphate level of four cloned puppies was significantly higher than the reference ranges. However, there was no significant difference among four cloned dogs. This study suggests that cloned puppies derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological and serological parameters.

Studies for the Processing Factors of Pesticides during the Milling of Wheat Grain (밀의 제분에 따른 밀가루 중 농약 가공계수 산출 연구)

  • Park, So-Young;Park, Kun-Sang;Im, Moo-Hyeog;Choi, Hoon;Chang, Moon-Ik;Kwon, Chan-Hyeok;Kim, Sun-Gu;Lee, Hyo-Ku;Hong, Moo-Ki;Shim, Jae-Han;Kim, Jeong-Han
    • The Korean Journal of Pesticide Science
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    • v.13 no.2
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    • pp.70-78
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    • 2009
  • Processing factors of pesticides in milling process of wheat grain, which are consumed and imported at large quantity were examined to establish reasonable MRL of the processed food. Azinphos-methyl, chlorpyrifos, chlorpyrifos-methyl, fenitrothion, malathion, and trichlorfon were selected for the study according to annual usage and the previous detection record in wheat grain. Dipping process for pesticide application was performed in laboratory, while milling process was conducted under pilot plant system. Processing factors were calculated by analyzing residual pesticides on wheat grain and processing products as wheat flour, bran and red dog. Processing factors were 0.05 for azinphos-methyl, 0.06 for chlorpyrifos, 0.05 for chlorpyrifos-methyl, 0.07 for fenitrothion, 0.07 for malathion, 0.06 for trichlorfon, respectively. Recovery test was also performed to establish extraction efficiency of analytical procedure. The recovery value ranged from 93.2% to 98.6% with standard deviation of 0.1-0.9%.

A Retrospective Study of Primary Glaucoma in Dogs: 43 cases (2006 ~ 2009) (개에서 발생한 원발성 녹내장 43례 (2006 ~ 2009))

  • Park, Young-Woo;Jeong, Man-Bok;Park, Shin-Ae;Kim, Won-Tae;Kim, Se-Eun;Ahn, Jae-Sang;Seo, Kang-Moon
    • Journal of Veterinary Clinics
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    • v.29 no.1
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    • pp.38-42
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    • 2012
  • To determine the prevalence of the primary glaucoma (PG) and occurrence of primary glaucoma in initially non-affected normal eye. Medical records of 7751 dogs presented to the Veterinary Medical Teaching Hospital (VMTH) of Seoul National University (SNU) between January 2006 and December 2009 were examined. Among 7751 dogs, forty three dogs (0.55%) developed PG. The following breeds were included in the study: 18 American Cocker Spaniels, 17 Shih Tzus, 3 Mixed Breeds, 2 Malteses, 2 Pekingeses, and 1 Chow Chow. The mean onset age of PG in the first eyes was $7.1{\pm}2.3$ years in the all breeds. Compared with Mixed Breed (0.54%), American Cocker Spaniel (3.16%, p = 0.004) and Chow Chow (20%, p = 0.001) had a higher risk of developing glaucoma. Twenty six of 43 dogs (60.1%) were females with male-to-female ratio of 1:1.53. Twenty nine of 33 eyes (87.9%) developed glaucoma in the fellow eye within mean ${\pm}SD$ time of $17.5{\pm}2.7$ months. From this study, American Cocker Spaniel and Shih Tzu were the most frequent PG breed in Korea. Also, unilateral PG dogs developed glaucoma in the non-affected normal eye within approximately 17 months. The results indicate that prophylactic medical therapy with antiglaucoma agents should be considered to delay or prevent developing glaucoma in non-affected normal eye.

Accuracy of CT image in measuring the mandible for implant : Effect of mandibular position and gantry angle (임플랜트를 위한 하악골 측정시 전산화단?사진상의 정확도에 관한 연구 : 하악 위치와 gantry각이 미치는 영향)

  • Choi Soon-Chul;Choi Hang-Moon;Park Rae-Jeong;Lee Sam-Sun;Park Tae-Won;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.28 no.1
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    • pp.225-234
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    • 1998
  • We used five adult dog mandibles embedded in resin block and six different cross-sectional planes for each mandible were choosen. According to the angle of mandibular occulsal plane to vertical plane(mandibular angle) and gantry angle of CT machine, we classified 4 experimental groups and 1 control group. The control group images were taken at the mandibular angel 0° and gantry angle 0°. The experimental images were taken at the mandibular angle 15° and gantry angle 0°(group 1); 30° and 0°(group 2); 15° and 15°(group 3) ;30° and 30°(group 4), respectively. Using the reformatted cross-sectional images, the distance from the mandibular canal to the alveolar crest and the distance from the mandibular canal to the buccal cortex and to the lingual cortex was measured and compared. The obtained results were as follows: 1. The distance from the mandibular canal to the alveolar crest of group 1 and 2 was larger than control group, but the distance of group 3 and 4 was smaller. The distance from the mandibular canal to the buccal cortex and to the lingual cortex of all experimental groups was smaller than control group. 2. The distance from the mandibular canal to the alveolar crest showed the largest difference from control group in all experimental groups, especially in group 2 and 4(p<0.05). 3. In the distance from the mandibular canal to the alveolar crest, the number of deviation value under 1 mm was 20 in group 3 and was 11 in group 2 and 4, respectively. 4. The deviation value of the distance from the mandibular canal to the buccal cortex and to the lingual cortex was under 1 mm in most cases.

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INFLUENCE OF CENTRAL PANORAMIC CURVE DEVIATION ON THE MANDIBULAR IMAGE RECONSTRUCTION IN THE IMPLANT CT (임플랜트전산화단층촬영시 CENTRAL PANORAMIC CURVE의 변화가 하악골의 영상 재구성에 미치는 영향)

  • Park Rae-Jeong;Lee Sam-Sun;Choi Soon-Chul;Park Tae-Won;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.28 no.1
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    • pp.47-58
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    • 1998
  • The purpose of this study was to investigate an influence of the change of central panoramic curves on the image reconstruction in the dental implant CT. The author designed three experimental groups according to the location of central panoramic curve. In group A, central panoramic curve was determined as the curve connecting the center of roots from the first premolar to the first molar. In group B, central panoramic curve was determined as the line connecting the lingual cortical plate at the level of the mesial aspect of the first premolar with the buccal cortical plate at the level of the mesial aspect of the first molar. In Group C, central panoramic curve was determined as the line connecting the buccal cortical plate at the level of the mesial aspect of the first premolar with the lingual cortical plate at the level of the mesial aspect of the first molar. Twenty four reformatted CT images was acquired from four mandibles embedded in the resin block and twenty four contact radiographs of dog specimens were acquired. Each Image was processed under Adobe Photoshop program analysed by MSPA(mandible/maxilla shape pattern analysis) variables such as MXVD, MXHD, UHD, MHD, and LHD. The obtained results were as follows ; 1. The mean of MXVD variable was 19.9, 20.2, and 20.0 in group A, B, and C, respectively, which were smaller than actual value 20.5. But, there was no significant difference among 3 groups (p>0.05). 2. The mean of MXHD, UHD, MHD, and LHD variables in group A, B, and C was 11.9, 12.2, and 12.3; 9.3, 9.5, and 9.6; 10.0, 10.3, and 10.3; 9.2, 9.3, and 9.4 respectively which were equal to or greater than the actual value 11.8, 9.3, 10.0, and 9.2. But, there was no significant difference among 3 groups (p>0.05). 3. The number of noneffective observations with difference over or under 1 mm with comparison to the actual value was 24(20%), 58(48.3%), and 52(43.3%), respectively, in group A, B, and C. 4. In group A, the number of observations over 1 mm and under 1 mm was 9 and 15, respectively, but in group Band C, the number of observations over 1 mm was more than under 1 mm.

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Feline adipose tissue-derived mesenchymal stem cells pretreated with IFN-γ enhance immunomodulatory effects through the PGE2 pathway

  • Park, Seol-Gi;An, Ju-Hyun;Li, Qiang;Chae, Hyung-Kyu;Park, Su-Min;Lee, Jeong-Hwa;Ahn, Jin-Ok;Song, Woo-Jin;Youn, Hwa-Young
    • Journal of Veterinary Science
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    • v.22 no.2
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    • pp.16.1-16.13
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    • 2021
  • Background: Preconditioning with inflammatory stimuli is used to improve the secretion of anti-inflammatory agents in stem cells from variant species such as mouse, human, and dog. However, there are only few studies on feline stem cells. Objectives: This study aimed to evaluate the immune regulatory capacity of feline adipose tissue-derived (fAT) mesenchymal stem cells (MSCs) pretreated with interferon-gamma (IFN-γ). Methods: To assess the interaction of lymphocytes and macrophages with IFN-γ-pretreated fAT-MSCs, mouse splenocytes and RAW 264.7 cells were cultured with the conditioned media from IFN-γ-pretreated MSCs. Results: Pretreatment with IFN-γ increased the gene expression levels of cyclooxygenase-2, indoleamine 2,3-dioxygenase, hepatocyte growth factor, and transforming growth factor-beta 1 in the MSCs. The conditioned media from IFN-γ-pretreated MSCs increased the expression levels of M2 macrophage markers and regulatory T-cell markers compared to those in the conditioned media from naive MSCs. Further, prostaglandin E2 (PGE2) inhibitor NS-398 attenuated the immunoregulatory potential of MSCs, suggesting that the increased PGE2 levels induced by IFN-γ stimulation is a crucial factor in the immune regulatory capacity of MSCs pretreated with IFN-γ. Conclusions: IFN-γ pretreatment improves the immune regulatory profile of fAT-MSCs mainly via the secretion of PGE2, which induces macrophage polarization and increases regulatory T-cell numbers.

Studies on Frozen Semen Characteristics Following Pentoxifylline Treatment and Artificial Insemination in Dog (개에서 Pentoxifylline 첨가에 따른 동결정액 성상과 인공수정에 관한 연구)

  • Ji, D.Y.;Kim, C.K.;Lee, J.H.;Park, S.J.;Ryu, L.S.;Ryu, J.W.;Lee, J.H.;Jeong, Y.C.;Pang, M.G.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.925-936
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    • 2005
  • The present study evaluated whether pentoxifylline added to the freezing extender could improve the sperm characteristics and function in canine frozen semen. Also the conception rate following AI with frozen-thawed semen was investigated. The beneficial effects of pentoxifylline supplementation were visible in motility, viability, acrosome reaction, and tail swelling patterns. Especially, highest sperm viability and function were obtained in the forzen semen supplemented with 1mM pentoxifylline. The follicle size measured by ultrasonography was 6.48 mm, 11.52 mm and 8.9 mm on 11, 13 and 15 days after the onset of natural estrus, respectively and ovulation occurred on 13 and 15 days. The pregnancy rates in bitches inseminated with frozen semen on natural and induced estrus were 71.4% and 75.0%, respectively. There was no significant difference between the pregnancy rates in bitches inseminated with frozen semen following natural and induced estrus, but the litter size was slightly increased in natural cycle.

A Short-Term Study of the Effects of UDCA on Gingival Inflammation in the Beagle Dog (우로수데옥시콜릭산이 치주질환 억제에 미치는 영향)

  • Park, Sang-Hyun;Han, Seoung-Min;Choi, Sang-Mook;Ku, Young;Rhyu, In-Chul;Han, Soo-Boo;Lee, Hak-Mo;Kim, Moon-Moo;Kim, Sang-Nyun;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.29 no.1
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    • pp.1-14
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    • 1999
  • Ursodeoxycholic acid(UDCA) is a hydrophilic gall bladder acid and has been used as a effective drug for liver disease related to in1munity. This drug inhibits secretions of IL-2, IL-4, and $IFN-{\gamma}$ from T-cells and production of immunoglobulin from B-cells. Also it has been reported that UDCA inhibits production of IL-1 related to the progression of periodontal disease and activation of collagenases. The purpose of the present study was to elucidate the effects of UDCA on inhibition of periodontal disease progression using clinical, microbiological and histometrical parameters. Twelve pure bred, 16 month-old-beagle dogs were used in the study. After ligature-induced periodontal diseases were formed, experimental drugs were applied twice a day and then the results of clinical, microbiological, and histometrical parameters were measured at baselie(initiation of experiment) , 4weeks and 8weeks. The gel with UDCA(concentration 0.5%, 5% 3 dogs in each) was applied to experimental group, chlorhexidine to positive control group(3dogs) and the gel without UDCA(base) to negative control group. After induction of general anesthesia, the maxillary 2nd, 3rd premolars and 1st molar and the mandibular 2nd, 3rd, 4th premolars and 1st molar were ligated in one side selected randomly and were not ligated in the opposite side. The plaque index(PI), gingival index(GI), pocket depth(PD) and gingival crevicular fluid(GCF) volum were measured clinically. The PI and GI were measured at 3 buccal points of all experimental teeth and the GCF was measured only at the 3rd premolar in the maxilla and the 4th premolar in the mandible. In the microbiological study, the samples extracted from the 3rd premolar of the maxilla and the 4th premolar of the mandible at the center of buccal surface were analyzed aerobics, anaerobics and Streptococcus colony forming units, After clinical and microbiological examination at 8weeks, the dogs were sacrificed by carotid artery perfusion. The samples were fixed and sectioned including interproximal area, and the distance from cementoenamel junction(CEJ) to alveolar crest was measured. The results were that PI, GI and PD increased until 4 weeks and decreased at 8 weeks in three groups but the differences between all the groups were not significant. The 0.5% UDCA in non-ligated group showed remarkable decrease of GCF. The experimental group applied 5% UDCA decreased the number of aerobics and anaerobics. The distance from CEJ to alveolar crest was greater in the negative control group on both ligated and non-ligated sides, but the differences were not significant stastically.

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