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In Vitro Development and Apoptosis in Haploid, Diploid Parthenotes and Fertilized Embryos

  • Y. J Chung;Lee, H. Y.;S. H. Jun;X. S. Cui;Kim, N. H.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.28-28
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    • 2003
  • Haploid parthenotes have been shown to be developmentally delayed compared with diploid parthenogenetic embryos in the mouse and pig. These developmental defects have been hypothesized to rusult from insufficient parthenogenetic activation, suboptimal in vitro culture conditions, or genemic imprinting. In the present study we compared the incidence of apoptosis and apoptosis related gene expression in pig haploid, diploid parthenotes and fertilized embryos. In vitro matured porcine oocytes were activated by electrical stimulation. Haploid activated oocytes with two polar bodies under stereomicroscopy were defined haploid parthenotes, oocytes with one polar body were defined as diploid parthenotes after 3h cycloheximide teatment. The morphological analysis of apoptosis in embryos was carried out using propidium iodide staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. The expression of Bcl-xL, Bak and P53 in haploid, diploid and in vivo fertilized blastocysts was determined using RT-PCR. Lower number of the haploid pig parthenotes developed to the morulae and blastocysts compared to the diploid parthnotes. Number of cells significantly lower in the haploid-derived blastocysts than diploid-derived it. Developmentally retarded haploid parthenotes exibited apoptosis at a significantly higher frequency than did diploid parthenotes and fertilized embryos. Level of Bcl-xL expression, diploid parthenotes similar to in vivo-derived it was higher than haploid parthenotes. However, Bak and P53 mRNA expression were not different among haploid, diploid, and fertilized embryos. This result suggested that parthenogenetic activation and parthenogenesis themselves do not cause apoptosis, but haploid increases the incidence of apoptosis in preimplantation embryos. Apoptosis may be due to decrease expression of Bcl-xL in haploid parthenotes developing in vitro.

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Cholesterol side-chain cleavage enzyme deficiency caused by a novel homozygous variant in P450 sidechain cleavage enzyme gene (CYP11A1) in a 46,XX Korean girl

  • Ye Ji Kim;Sun Cho;Hwa Young Kim;Young Hwa Jung;Jung Min Ko;Chang Won Choi;Jaehyun Kim
    • Journal of Genetic Medicine
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    • v.20 no.1
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    • pp.25-29
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    • 2023
  • The CYP11A1 gene encodes for the cholesterol side-chain cleavage enzyme (P450scc), which initiates steroid hormone biosynthesis. Defective P450scc activity results in severe glucocorticoid and mineralocorticoid deficiencies. We describe a case of P450scc deficiency due to a novel homozygous CYP11A1 variant inherited from the mother with a possibility of uniparental disomy (UPD). The patient was a female, had no family history of endocrine disease, and showed adrenal insufficiency at 13 days of age. Hormonal analysis with an adrenocorticotropic hormone stimulation test showed both glucocorticoid and mineralocorticoid deficiencies, presumed to be a defect of the early stage of steroidogenesis. Exome sequencing reported a novel homozygous frameshift variant of CYP11A1 (c.284_285del, p.Asn95Serfs*10), which was inherited from the mother. Additionally, homozygosity in 15q22.31q26.2, which included CYP11A1, was identified using a chromosomal microarray. It was suggested that the possibility of maternal UPD was involved as the cause of a P450scc deficiency by unmasking the maternally derived affected allele. To our understanding, P450scc deficiency associated with UPD encompassing CYP11A1 had not been reported in Korea before. Genetic analysis can help diagnose rare causes of primary adrenal insufficiency, including P450scc deficiency.

Expression of Senescence-Associated Secretory Phenotype in Senescent Gingival Fibroblasts

  • Sangim Lee
    • Journal of dental hygiene science
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    • v.23 no.2
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    • pp.169-175
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    • 2023
  • Background: Although microbial infection is direct cause of periodontal disease, various environmental factors influence the disease severity. Aging is considered a risk factor for oral diseases, with the prevalence of periodontal diseases increasing with age. Moreover, senescence-associated secretory phenotype (SASP) expressed in age-related diseases is a key marker of chronic inflammation and aging phenotypes. Therefore, this study aimed to understand the relevance of senescent cells to periodontal health and disease, investigate the possibility of regulating the expression of aging- and osteolysis-related factors in gingival fibroblasts, and investigate the effect of senescence induction in gingival fibroblasts on osteoclast differentiation in mouse bone marrow-derived macrophages (BMMs). Methods: After stimulation with 400 nM hydrogen peroxidase, human gingival fibroblasts (HGFs) were examined for senescence-associated β-galactosidase. Western blot and enzyme-linked immunosorbent assays were performed to assess the expression of SASP. Osteoclast formation was assessed in BMMs using a conditioned medium (CM) from hydrogen peroxide-stimulated HGFs. Osteoclastic differentiation was investigated using tartrate-resistant acid phosphatase (TRAP) staining and activity. Data analysis was performed using SPSS version 25.0. Results: The expression of senescence-related molecules, including p53, p16, and p21, and the expression of osteolytic factors, including IL-6, IL-8, and IL-17, were found to be significantly higher in the hydrogen peroxide-stimulated HGF than in the control group. Regarding the indirect effects of senescent gingival cells, the number of osteoclasts and TRAP activity increased according to the differentiation of BMM cultured in CM. Conclusion: Our results on the of between osteolytic factors and cellular senescence in gingival fibroblast cells helped to reveal evidence of pathological aging mechanisms. Furthermore, our results suggest that the development of novel therapies that target specific SASP factors could be an effective treatment strategy for periodontal disease.

Recombinant Human HAPLN1 Mitigates Pulmonary Emphysema by Increasing TGF-β Receptor I and Sirtuins Levels in Human Alveolar Epithelial Cells

  • Yongwei Piao;So Yoon Yun;Zhicheng Fu;Ji Min Jang;Moon Jung Back;Ha Hyung Kim;Dae Kyong Kim
    • Molecules and Cells
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    • v.46 no.9
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    • pp.558-572
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    • 2023
  • Chronic obstructive pulmonary disease (COPD) will be the third leading cause of death worldwide by 2030. One of its components, emphysema, has been defined as a lung disease that irreversibly damages the lungs' alveoli. Treatment is currently unavailable for emphysema symptoms and complete cure of the disease. Hyaluronan (HA) and proteoglycan link protein 1 (HAPLN1), an HA-binding protein linking HA in the extracellular matrix to stabilize the proteoglycan structure, forms a bulky hydrogel-like aggregate. Studies on the biological role of the full-length HAPLN1, a simple structure-stabilizing protein, are limited. Here, we demonstrated for the first time that treating human alveolar epithelial type 2 cells with recombinant human HAPLN1 (rhHAPLN1) increased TGF-β receptor 1 (TGF-β RI) protein levels, but not TGF-β RII, in a CD44-dependent manner with concurrent enhancement of the phosphorylated Smad3 (p-Smad3), but not p-Smad2, upon TGF-β1 stimulation. Furthermore, rhHAPLN1 significantly increased sirtuins levels (i.e., SIRT1/2/6) without TGF-β1 and inhibited acetylated p300 levels that were increased by TGF-β1. rhHAPLN1 is crucial in regulating cellular senescence, including p53, p21, and p16, and inflammation markers such as p-NF-κB and Nrf2. Both senile emphysema mouse model induced via intraperitoneal rhHAPLN1 injections and porcine pancreatic elastase (PPE)-induced COPD mouse model generated via rhHAPLN1-containing aerosols inhalations showed a significantly potent efficacy in reducing alveolar spaces enlargement. Preclinical trials are underway to investigate the effects of inhaled rhHAPLN1-containing aerosols on several COPD animal models.

The Analysis of Research Trend about Management of Low Back Pain (요통관리에 관한 연구동향 분석)

  • Hyun, Kyung-Sun
    • The Korean Journal of Rehabilitation Nursing
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    • v.1 no.1
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    • pp.51-60
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    • 1998
  • The result of studying papers about management of lower back pain published in domestic and foreign nursing and medical magazines in these 10 years is as follows ; 1. General characteristic of lower back pain ; 1) In sex distribution, there were more men than women in 5 papers and more women than men in 4 papers among 9 papers surveryed. 2) In age distribution, thirties to forties of patients had more lower back pain as surveyed by general hospitals and fifties to sixties of patients had more lower back pain as surveyed by oriental medical hospitals. 3) In cause factor, there were 50 to 65% of sprain, 32 to 44% of herniated intervertebral disc and 13 to 29% of degenerative changes. 4) In symtom distribution, there were 26 to 57% of lower back pain, 42 to 65% of lower back pain with radiating pain and 34 to 99% of paravertevral muscle spasm. 5) In period of pain management distribution, 18 to 40% of patients experienced pain for less than 6 months and 59 to 82% of them experienced pain for more than 6 months in 3 papers among 4 papers. 6) In surveying the treatment, 66 to 88% of patients had conservative treatment and there were treatments of general hospital, oriental medicine, self remedy and traditional practice in conservative treatment. 7) In job distribution, 12 to 50% of them were housekeepers, 23 to 31% office workers, 4.6 to 36% blue color workers and 11 to 15% students. 2. As psychological character lower back pain paients had anxiety, depression, anger-hostility, phobic anxiety, neurasthenia, hypochondriasis, and interpersonal sensitivity. 3. To distinguish the cause of lower back pain, plain lumbar roentgenogram, straight leg rasing test, eletromyelogic findings, somatosensory evoked potentials CT and MRI were performed. 4. To relieve lower back pain. epidural adhesiolysis, epidulal injection of local anesthetic in mixture with steroid, lumbar spinal root block, low level laser therapy, acupuncture like transcutaneous nerve stimulation(AL TENS), topical capsaicin and lumbar orthotics were used in medical field, and relaxation technique was used in nursing field. 5. Mckenzie's extension exercise and William's flexsion exercise for lower back pain were used in medical field and Yoga exercise was applied in nursing field. 6. The more school education and self efficacy were high, the better they had active coping lower back pain positively and the less self efficacy was the more they had serious pain. As a result of studying the paper there have been very little research for lower back pain in nursing fields of Korea and foreign countries. Because 60 to 80% of population expeience lower back pain at least more than once, it is necessary to develop the study and clinical practice for management of lower back pain.

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Healing Effect Assessment by the Autonomic Nervous Responses Using the Aroma-Treated Fabrics (자율신경반응에 의한 방향성 의류소재의 힐링효과 평가)

  • Park, Hae-Li;Cho, Gil-Soo
    • Science of Emotion and Sensibility
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    • v.18 no.2
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    • pp.19-30
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    • 2015
  • The healing effect of the aroma treated fabrics with lavender and lemon aromas was investigated by assessing the autonomic nervous responses of human body. For this cause Lemon and lavender microcapsules were coated on a cotton fabric using a water-based acrylic binder, respectively. And the study created a total of four aroma treated fabrics at a concentration of 2% and 5% respectively. Electrocardiogram(ECG), skin conductance, and Blood flow, of ten participants were measured for 30 sec at a stable condition, at a stress status (working memory task), and at a stimulation status (after rubbing aroma treated fabrics). Subjective sensibilities of the aromas were also evaluated. With regard to the responses of the autonomic nervous system, in order to understand how the values gained after the normalization process would cause different physiological signals between the stable state and the aroma-stimulated state as well as between the stress state and the aroma-stimulated state, the study conducted a non-parametric test, friedman test as well and analyzed tendencies. LF/HF turned out to be significantly different to the stress state, and according to the results of the post-hoc comparison, lemon 5% presented statistically significant differences among the lavender 2%, lavender5%, lemon2%. Lemon 5% stimuli increased stress but stimuli consisting of the lavender 2%, the lavender 5% and the lemon 2% decreased stress because of a psychological rest. And the stimuli of the lavender 2%, the lavender 5%, the lemon 2% presented a healing effect in this research.

Analysis on the Mismatch between Instructional Design and Teaching Practice of Pre-service Science Teachers in Teaching Practicum (교육실습에서 중등 예비과학교사들의 수업계획과 실제수업의 불일치 분석)

  • Jung, Juwon;Lee, Bongwoo
    • Journal of The Korean Association For Science Education
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    • v.36 no.3
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    • pp.435-443
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    • 2016
  • The purpose of this study is to analyze the mismatch between instructional design and teaching practice of pre-service science teachers and to investigate the cause of these mismatches. Twenty pre-service science teachers took part in teaching practicum for four weeks from Apr. 2015 to May 2015 and we analyzed their lesson plans and videos of these lessons. After that, we interviewed the pre-service science teachers in order to know the cause of these mismatches and additional informations. The main findings are as follows. First, in the introductory stage, we found more mismatch in the "attention and motivation stimulation" area than any other area. Many pre-service science teachers corrected their 'motivation practice' in different forms. Second, we found out that the most mismatch occurred in the evolving stage. Many pre-service science teachers added learning contents in "learning content presentation" area. Third, in the closing stage, many pre-service science teachers omitted the "leaning content summary" area in the lack of time. Fourth, the number of mismatches by internal factors is similar of by external factors. The mismatches by external factors were mainly by feedback of guidance teacher and change by students' response. In addition, we discussed the implications related to reflective mentoring program, importance of guidance teacher, importance of time management etc.

PM2.5 in poultry houses synergizes with Pseudomonas aeruginosa to aggravate lung inflammation in mice through the NF-κB pathway

  • Li, Meng;Wei, Xiuli;Li, Youzhi;Feng, Tao;Jiang, Linlin;Zhu, Hongwei;Yu, Xin;Tang, Jinxiu;Chen, Guozhong;Zhang, Jianlong;Zhang, Xingxiao
    • Journal of Veterinary Science
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    • v.21 no.3
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    • pp.46.1-46.18
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    • 2020
  • Background: High concentrations of particulate matter less than 2.5 ㎛ in diameter (PM2.5) in poultry houses is an important cause of respiratory disease in animals and humans. Pseudomonas aeruginosa is an opportunistic pathogen that can induce severe respiratory disease in animals under stress or with abnormal immune functions. When excessively high concentrations of PM2.5 in poultry houses damage the respiratory system and impair host immunity, secondary infections with P. aeruginosa can occur and produce a more intense inflammatory response, resulting in more severe lung injury. Objectives: In this study, we focused on the synergistic induction of inflammatory injury in the respiratory system and the related molecular mechanisms induced by PM2.5 and P. aeruginosa in poultry houses. Methods: High-throughput 16S rDNA sequence analysis was used for characterizing the bacterial diversity and relative abundance of the PM2.5 samples, and the effects of PM2.5 and P. aeruginosa stimulation on inflammation were detected by in vitro and in vivo. Results: Sequencing results indicated that the PM2.5 in poultry houses contained a high abundance of potentially pathogenic genera, such as Pseudomonas (2.94%). The lung tissues of mice had more significant pathological damage when co-stimulated by PM2.5 and P. aeruginosa, and it can increase the expression levels of interleukin (IL)-6, IL-8, and tumor necrosis factor-α through nuclear factor (NF)-κB pathway in vivo and in vitro. Conclusions: The results confirmed that poultry house PM2.5 in combination with P. aeruginosa could aggravate the inflammatory response and cause more severe respiratory system injuries through a process closely related to the activation of the NF-κB pathway.

Regulatory Mechanism of Lipopolysaccharide(LPS)-Induced Interleukin-8 Gene Expression in Mononuclear Phagocytic Cells (단핵식세포에서 내독소에 의한 인터루킨-8 유전자 발현 조절기전에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.462-474
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    • 1994
  • Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.

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Differences on Use of Colors before and after Color Stimulations -Two different stimuli of images by design majors and non-design majors- (색채자극 전과 후 달라진 색 사용에 관한 연구)

  • Kim, Sun-Hwa;Lee, Seung-Hee
    • Science of Emotion and Sensibility
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    • v.12 no.3
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    • pp.351-356
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    • 2009
  • The study is about how the difference in education levels can affect the subjects' use of color and find out the features of the effect. This experimental study focused on the differences in color expression using the outline picture of a painting before and after being exposed to a full-color picture of the original painting. Differences between two conditions with two different stimuli were observed and the cause of the differences was analyzed based on the number of colors and the quantity of colors and composition of the colors used by the subjects. The study precedes an experiment based on how the level of education though experience can affect the use of colors. This study is focused on the various effects of color usage by children and adults who have various degrees of visual experience. In addition, the study analyzes how design majored subject differed in the use of color from non-design majored subject and compares the use of colors by the two groups. The results showed that design majored subject tended to use less coloring patterns after seeing the stimulus of the Monet picture with more colors, and included the colors from the stimuli they were exposed to. The non-design majored subject who viewed the Mondrian painting demonstrated opposite results by using less number of colors than before seeing the stimuli. In comparison to design majored subject, the variations in selected colors and the quantity of color by non-design majored subject were not consistent.

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