• Journal of Veterinary Science
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• v.19 no.6
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• pp.759-770
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• 2018

Fowl adenovirus (FAdV) is distributed worldwide and causes economic losses in the poultry industry. The objectives of this study were to determine the hexon and fiber gene changes in an attenuated FAdV isolate from Malaysia in specific pathogen-free chicken embryonated eggs (SPF CEE) and its infectivity in commercial broiler chickens. SPF CEE were inoculated with 0.1 mL FAdV inoculum via the chorioallantoic membrane (CAM) for 20 consecutive passages. The isolate at passage 20 (E20), with a virus titer of $10^{8.7}TCID_{50}/mL$ ($TCID_{50}$, 50% tissue culture infective dose), was inoculated (0.5 mL) into one-day-old commercial broiler chicks either via oral or intraperitoneal routes. The study demonstrated that 100% embryonic mortality was recorded from E2 to E20 with a delayed pattern at E17 onwards. The lesions were confined to the liver and CAM. Substitutions of amino acids in the L1 loop of hexon at positions 49 and 66, and in the knob of fiber at positions 318 and 322 were recorded in the E20 isolate. The isolate belongs to serotype 8b and is non-pathogenic to broiler chickens, but it is able to induce a FAdV antibody titer. It appears that molecular changes in the L1 loop of hexon and the knob of fiber are markers for FAdV infectivity.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.1
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• pp.2-11
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• 2024

Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

• Tuberculosis and Respiratory Diseases
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• v.53 no.5
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• pp.569-576
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• 2002

Background : Chlamydia pneumoniae is one of common causes in upper and lower respiratory infections. Isolating C. pneumoniae from clinical specimens is very difficult due to the characteristics of the organism. Recently, we succeeded in isolating C. pneumoniae from a Korean patient, who suffered from acute pharyngitis. This is the first isolate from a clinical specimen in Korea. Methods : We attained a nasopharyngeal swab from a 22-year-old female patient, and inoculated it on a monolayer of the Hep-2 cell line. After 8 passages, we found the inclusion bodies of C. pneumoniae by an immunofluorescence(IF) test. The species-specific monoclonal antibody IF staining and species-specific PCR were done to confirm the species of the isolate, and electron microscopy was used to characterize the morphology. Results : The isolated was confirmed to be C. pneumoniae by species-specific IF and PCR, and the strain was named LKK-1. The shape of the elementary body was round and with a narrow periplasmic space, as shown by electron microscopy, which is similar to the Japanese strain, but not the Western strain. Conclusion : We succeeded in isolating C. pneumoniae from a 22-year-old patient with acute pharyngitis, which is the first isolate in Korea. In the future, this Korean strain will be useful to the study of C. pneumoniae.

• Journal of Life Science
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• v.18 no.3
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• pp.344-349
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• 2008

Normal somatic cells proliferate for a limited number of doublings in culture and then enter an irreversible growth-arrest state called replicative senescence. Replicative senescence has been believed a reason for the limited cellular turnover and deterioration of tissue function in aged animals. However, there is no experimental evidence supporting this assumption. Furthermore, cells from aged person have been poorly characterized with an exception of the cases of T cells. In this study, we examined cell biological changes occurring in replicative senescence of fibroblast strains originated from a new-born (NHF-NB) and a 87 year old man (NHF-87). NHF-87 (and the cells from a 75-year old) proliferated to smaller population doublings and with longer doubling times than NHF-NB did. At early passages, NHF-87 exhibited a low senescence-associated ${\beta}-Gal$ (SA ${\beta}-Gal$) activity and lipofuscin level, typical markers for cellular senescence. Furthermore, they maintained low levels of lysosome and reactive oxygen species (ROS). All of these levels increased dramatically in the late passage NHF-87 quite similarly as those in the late passaged NHF-NB did. These results indicate that most cells originated from the aged maintain a phenotype of the cells originated from new-born donors and undergo replicative senescence with the same kinetics as that of the cells from new-born. It is also indicated that not SA ${\beta}-gal$ activity but cell proliferation rate may be qualified as a biomarker for cells aged in vivo.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.387-390
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• 1995

In the present communication, YS-27, a Korean strain of Entnnloeba histoIWtica is described for the isolation and establishment of axonic cultivation. 5. histoLvticc, designated as strain "YS-27" was isolated from the pus of a hepatic abscess obtained from a 72 you old inpatient of August 10, 1969. Specimens, were obtained by needle aspiration, inoculated immediately and weekly cultured in a modified diphasic medium at 37℃. Strain YS-27 had been maintained for more than 15 years by weekly subculture until February, 1985. These cultures were transferred to a monoxenic TTY-SB medium seeded with a trypanosomatid of the genus CyithidiG. Penicillin G, 2 to 10 H 103 International units and Streptomycin, 2 to 10 mg per 100 ml, were added to the cultures to eliminate the bacteria. After more than one year later, these two organisms were well maintained by transfer every 3 or 4 days until .January. 1986 at 37℃ in TTY-SB medium in the absence of other microorganisms. These monoxenic cultures were then transferred to TYI-S-33 medium. Strain YS-27 alone had not been growing at the time of transfer, but when overlaid with Crithinia at intervals of 3 to 4 days, strain YS-27 propagated well. The Clthidio died out several weeks later after several passages. Beginning in April, 1986, strain YS-27. was successfully established in axonic culture in TYI-S-33 medium and has been maintained in continuous culture and multiplied well to present.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.179-184
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• 2007

Culture method of somatic cells is one of the important factors in the production of transgenic pigs by somatic cell nuclear transfer. In this study, we established an efficient culture method of porcine fetal fibroblasts. Porcine fetal fibroblasts were isolated from 33-day-old fetuses. The proliferation of porcine fetal fibroblasts was analyzed by different serum types and culture media. The cultures in medium supplied 15% ES screened FBS showed faster increase in cell number than 15% FBS. Also, fetal fibroblasts have been propagated continuously for $7{\sim}8$ passages in ES modified DMEM and DMEM medium. We transfected $PGK-neo^r$ vector (pKJ2) into porcine fetal fibroblasts to estimate colony formation in this culture condition. The formation of colonies was confirmed in the medium containing $300\;{\mu}g/ml$ G418 at 12 day. These data show that this culture system can be used screening of porcine somatic cells transfected transgene.

• Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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• v.17 no.2
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• pp.98-102
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• 2006

Background and Objectives : Voice onset time(VOT) is defined as the time interval from oral release of a stop consonant to the onset of glottal pulsing in the following vowel. VOT is a temporal characteristics of stop consonants that reflects the complex timing of glottal articulation relative to supraglottal articulation. Stop consonants are characterized by creation of a pressure difference across a complete occlusion in the vocal tract, followed by a sudden release 'burst' due to opening that occlusion. The objects of this study is to evaluate a usefulness of voice onset time in the assessment of voice disorderd patients. Subjects : Subjects were 20 adults with normal voice and with benign laryngeal disorders. Subjects with voice disorders represented the following vocal pathologies : vocal polyp, vocal nodule, Reinke's edema and unilateral vocal fold paralysis(UVFP). Control subjects were matched for age (21-40 yews old) and sex(male) with the voice disorders subjects and had normal vocal qualities with no history of voice disorders. Methods : Each voice-disordered and matched control subject read the test passages containing three types of Korean bilabial consonants. VOT measures were made for the initial $/p/p^h/\;and\;/p'/$. VOT was measured using acoustic waveform or wide band spectrogram. Results : For each voiceless stop consonants, there was a significant difference in VOT between the voice disordered and normal subjects. The mean VOTs of the lax stops in UVFP was significantly shorter than those of control subjects in the UVFP. The mean VOTs of the aspirated stops in the vocal polyp and nodule were longer than those of control subjects, but not significant. The mean VOTs of the glottalized in voice disordered groups were longer than those of control subjects, and significant statistically in the UVFP. Conclusions : VOT may be a clinically useful acoustic parameter in the assessment of voice disordered patients, especially in the unilateral vocal fold paralysis.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.243-248
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• 2003

Sperrnatogenesis, the process by which the male germ-line stem cells(GSCs; type A spermatogonia) divide and differentiate to produce the mature spermatozoa, occurs in the seminiferous tubules of the testis. The GSCs proliferate actively to produce two types of cells: other GSCs and differentiating spermatogonia. GSCs have unipotentcy, devoted solely to the generation of sperm. The function of GSCs has broad implications for development, disease, and evolution. Spermatogenesis is fundamental for propagation of species and the defects of this system can result in infertility or disease. The ability to identify, isolate, culture, and alter GSCs will allow powerful new approaches in animal transgenesis and human gene therapy relating to infertility. Until recently, research on stem cells in the testis has been limited because of technical difficulties in isolating and identifying these cell populations. Here, we were trying to find out optimal conditions for in vitro culture of GSCs for identifying and isolating GSCs. We collected mouse GSCs from 3-days old mouse by two-step enzyme digestion method. GSCs were plated and grown on mouse embryonic fibroblasts in Dulbecco's modified Eagle's medium (DMEM) containing 15％ fatal bovine serum, 10 mM 2-mercaptoethanol, 1％ non-essential amino acids, 1 ng/$m\ell$ bFGF, 10 $\mu$M forskolin, 1500 U/$m\ell$ human recombinant leukemia inhibitory factor (LIF). Over a period 3∼5 days, GSCs gave rise to large multicellular colonies resembling those of mouse pluripotent stem cells. After 5th passages, cells within the colonies continued to be alkaline phosphatase and Oct-4 positive and tested positive against a panel of two immunological markers(Integrin $\alpha$ 6 and Integrin $\beta$ 1) that have been recognized generally to characterize GSCs. SSEA-1, SSEA-3, and SSEA-4 also showed positive signals. Based on our data, these GSCs-derived cultures meet the criteria for GSCs itself and even other pluripotent stem cells. We reported here the establishment of in vitro cultures from mouse male GSCs.

• Journal of the korean academy of Pediatric Dentistry
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• v.48 no.3
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• pp.291-301
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• 2021

The aim of this study is to compare the properties of odontoblast gene of early passage cells and late passage cells derived from impacted maxillary supernumerary teeth. Impacted supernumerary teeth with maxilla were extracted from 12 patients (8 males, 4 females) between 6 - 9 years old without medical history. Real-time polymerase chain reaction (PCR) was conducted to compare characterization of odontoblast cell in the 3rd and 10th passage, and between with bone inducing additive group and without additive group. Genes for odontoblasts characteristics are osteonectin (ONT), alkaline phosphatase (ALP), osteocalcin (OCN), dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP). The level of gene expression was in a decreasing order of ONT, ALP, OCN, DMP-1 and DSPP in the 3rd passage, and in decreasing order of ONT, DMP-1, OCN, ALP, and DSPP in the 10th passage in the undifferentiation and differentiation group. The order of ONT, DMP-1, and OCN did not changed. ALP and DMP-1 were switched in order. ALP and DMP-1 may be used as important markers for differentiating between the 3rd passage and 10th passage cells. Considering that supernumerary tooth was extracted young age and the time required to cultured 10th passage was short, supernumerary tooth can be considered a useful donor site of dental pulp stem cells.

• Journal of the Daesoon Academy of Sciences
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• v.38
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• pp.47-82
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• 2021

At the age of 15, Song Gyu, the second patriarch of Won Buddhism, met Jeungsan-gyo members and was substantially influenced by them. Jeongsan cultivated himself for three months in Mount Gaya based on their recommendations. He instructed his family members to practice reciting the Tae-eul Mantra. Henceforth, Jeongsan was said to attain supernatural power when he was around 18 years old, and he pursued the traces left by Jeungsan in Jeolla Province. Once there, he asked Jeungsan's younger sister to move to his hometown, Seongju, Gyeongsang Province, and he served her with his utmost sincerity. He went back to the birthplace of Jeungsan and received a Daoist book from Jeungsan's daughter titled, Essentials for an Upright Mind (正心要訣). Jeongsan practiced holy works for 10 months at Daewon-sa Temple in Mount Moak where Jeungsan was said to have attained unification with the Dao. After he had met Jeungsan-gyo members at the temple, he was able to stay in her house where he ended up meeting So Taesan. Before their meeting, it is obvious that Jeongsan was a member of Jeungsan-gyo. Afterward, Jeongsan entered into Won Buddhism and used the passage, 'saving lives by curing the world (濟生醫世).' He recited the writing of Jeungsan, which had been given to his disciples, as if it had been a mantra. In addition, he mentioned Jeungsan's poems or the Chinese poems that he had quoted many times. Jeongsan also interpreted passages from The Hyunmu Scripture (玄武經) written by Jeungsan in a unique manner. Jeongsan answered his disciples in his own way when they asked questions on the teachings of Jeungsan. He recognized Jeungsan as one of the Three Primes, who presided over the Great Opening.