• Title/Summary/Keyword: Pax6

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Application of Immunophenotyping and Heteroduplex Polymerase Chain Reaction (hPARR) for Diagnosis of Canine Lymphomas

  • Sirivisoot, Sirintra;Techangamsuwan, Somporn;Tangkawattana, Sirikachorn;Rungsipipat, Anudep
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.2909-2916
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    • 2016
  • Background: Canine malignant lymphoma is classified into B- or T-cell origin, as in the human case. Due to differences in prognosis, a suitable method needs to be developed for lineage identification. Aims: To determine the accuracy of immunophenotypic and molecular information between three methods: immunocytochemistry (ICC), immunohistochemistry (IHC) and heteroduplex polymerase chain reaction for antigen receptor rearrangements (hPARR) in spontaneous canine lymphomas. Materials and Methods: Peripheral blood, fine needle aspiration and tissue biopsies from enlarged peripheral lymph nodes prior to treatment of 28 multicentric lymphoma patients were collected. Cytopathology and histopathology were examined and classified using the updated Kiel and WHO classifications, respectively. Anti-Pax5 and anti-CD3 antibodies as B- and T-cell markers were applied for immunophenotyping by ICC and IHC. Neoplastic lymphocytes from lymph node and white blood cell pellets from peripheral blood were evaluated by hPARR. Results: In this study, low grade B-cell lymphoma accounted for 25% (7/28), high grade B-cell lymphoma for 64.3% (18/28) and high grade T-cell lymphoma for 10.7% (3/28). According to the WHO classification, 50% of all cases were classified as diffuse large B-cell lymphoma. In addition, ICC showed concordant results with IHC; all B-cell lymphomas showed Pax5+/CD3, and all T-cell lymphomas exhibited Pax5-/CD3+. In contrast to hPARR, 12 B-cell lymphomas featured the IgH gene; seven presented the $TCR{\gamma}$ gene; five cases showed both IgH and $TCR{\gamma}$ genes, and one case were indeterminate. Three T-cell lymphomas showed the $TCR{\gamma}$ gene. The percentage agreement between hPARR and ICC/IHC was 60%. Conclusions: Immunophenotyping should not rely on a single method. ICC or IHC with hPARR should be used concurrently for immunophenotypic diagnosis in canine lymphomas.

Insect peptide CopA3 promotes proliferation and PAX7 and MYOD expression in porcine muscle satellite cells

  • Jeongeun, Lee;Jinryoung, Park;Hosung, Choe;Kwanseob, Shim
    • Journal of Animal Science and Technology
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    • v.64 no.6
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    • pp.1132-1143
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    • 2022
  • Insects are a valuable natural source that can produce a variety of bioactive compounds due to their increasing species diversity. CopA3 is an antimicrobial peptide derived from Copris tripartitus (i.e., the dung beetle). It is known to increase the proliferation of colonic epithelial and neuronal stem cells by regulating cell cycle. This research hypothesized that CopA3 can promote the proliferation of porcine muscle satellite cells (MSCs). The effects of CopA3 on porcine MSCs, which are important for muscle growth and regeneration, remain unclear. Here, we investigated the effects of CopA3 on porcine MSCs. According to viability results, we designed four groups: control (without CopA3) and three treatment groups (treated with 5,10, and 25 ㎍/mL of CopA3). At a CopA3 concentration of 5 ㎍/mL and 10 ㎍/mL, the proliferation of MSCs increased more than that observed in the control group. Furthermore, compared to that in the control, CopA3 treatment increased the S phase but decreased the G0/G1 phase ratio. Additionally, early and late apoptotic cells were found to be decreased in the 5 ㎍/mL group. The expressions of the myogenesis-related transcription factor PAX7 and MYOD proteins were significantly upregulated in the 5 ㎍/mL and 10 ㎍/mL groups, whereas the MYOG protein remained undetected in all group. This study suggested that CopA3 promotes muscle cell proliferation by regulating the cell cycle of MSCs and can regulate the activity of MSCs by increasing the expressions of PAX7 and MYOD.

Derivation of Neural Precursor Cells from Human Embryonic Stem Cells

  • Kim Sehee;Hong Ji Young;Joo So Yeon;Kim Jae Hwan;Moon Shin Yong;Yoon Hyun Soo;Kim Doo Han;Chung Hyung Min;Choi Seong-Jun
    • Reproductive and Developmental Biology
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    • v.28 no.4
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    • pp.247-252
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    • 2004
  • Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.

Wilms' tumor, aniridia, genitourinary anomalies, and mental retardation (WAGR) syndrome: Successful treatment of the first case with bilateral Wilms' tumors in Korea (윌름즈 종양, 무홍체증, 비뇨생식기계 기형, 정신지체 (WAGR) 증후군 : 양측성 윌름즈 종양을 성공적으로 치료한 국내 첫 증례 보고)

  • Min, Kyung Sun;Baek, Hee Jo;Han, Dong Kyun;You, Ju Hee;Hwang, Tai Ju;Kwon, Dong Deuk;Kook, Hoon
    • Clinical and Experimental Pediatrics
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    • v.51 no.12
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    • pp.1355-1358
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    • 2008
  • Wilms' tumor, aniridia, genitourinary anomalies, and mental retardation (WAGR) syndrome is caused by deletion of chromosome 11p13, including the Wilms' tumor (WT1) and aniridia gene (PAX6) loci. Here, we report the first case of WAGR syndrome in Korea; the patient was a 2-year-old girl with bilateral aniridia from birth who presented with abdominal distention and mental retardation. Cytogenetically, she had deletion of chromosome 11p11.2-13. Bilateral Wilms' tumors were successfully treated by chemotherapy and surgery. She has been tumor-free for 19 months off chemotherapy with preserved renal function.

Local Work-function Variation of the Initial Oxidation-Stages of Si(111)-7${\times}$7 (Si(111)-7${\times}$7 표면의 초기산화 단계에서의 국부 일함수 변화)

  • Im, Sam-Ho;Gu, Se-Jeong;Kim, Gi-Jeong;Park, Chan;Seo, Jae-Myeong
    • Journal of the Korean Vacuum Society
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    • v.2 no.2
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    • pp.166-170
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    • 1993
  • In the initial stages of oxidation of Si(111)-7${\times}$7 held at 40K through exposing molecular oxygen, it has been detected average work-function measured by ultraviolet photoelectron spectroscopy (UPS) is about 0.4eV higher than the local work-function of the modified area measured by photoemission for adsorbed Xenon (PAX). This result indicates that the increment of work-function at the initial oxidation stages of Si(111)-7${\times}$7 is mainly due to the moleculary adsorbed oxygen. From the shift of broadened Xe 5p and Xe 3d, it has also been estimated that the work-function of the modified area is 0.6eV higher than that of the clean area.

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Investigation of the effect of Blueberry hydrothermal extracts on myoblast differentiation (블루베리 열수 추출물의 근아세포의 근분화에 미치는 영향)

  • Choi, Youngsoo;Kim, Eunmi;Choi, Sunkyung;Lee, Woonghee;Han, Hyosang;Kim, Keekwang
    • The Korea Journal of Herbology
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    • v.35 no.3
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    • pp.25-32
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    • 2020
  • Objectives : At present, aging-related degenerative muscle diseases are considered a serious problem. However, the effects on muscles regarding the efficacy of blueberry have not been studied. In this study, we tried to find out the correlation between blueberry and muscle. Methods : 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay was performed to confirm the antioxidant efficacy of blueberry hydrothermal extract. To determine the effect of blueberry hydrothermal extracts (BHE) on myoblast activity, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed. To confirm the effect of blueberry hydrothermal extracts on the differentiation of myoblast into myotubes, protein expression levels of myosin heavy chain 3 (Myh3) and paired box 3/7 (pax3/7) were confirmed by immunoblot analysis. In addition, immunofluorescence microscopy was performed to confirm the effect on myotube formation of blueberry hydrothermal extracts. Results : Antioxidative efficacy and low toxicity were confirmed through ABTS assay and MTS assay of blueberry extract for myoblasts. As a result of immunoblot analysis and immunofluorescence analysis, the decrease in myogenic marker Pax3/7 was not confirmed, but myotubes The specific expression inhibitory activity of the forming protein Myh3 was confirmed. Through this, it was confirmed that the blueberry extract has a negative activity against myoblast differentiation. Conclusion : This experiment confirmed that blueberry hydrothermal extract has excellent antioxidant efficacy and negative results in inhibiting the differentiation and proliferation of myoblast. This requires deep study of certain ingredients and requires reassessment of the dietary intake of blueberries.

Genetic analysis using whole-exome sequencing in pediatric chronic kidney disease: a single center's experience

  • Lee, Hyeonju;Min, Jeesu;Ahn, Yo Han;Kang, Hee Gyung
    • Childhood Kidney Diseases
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    • v.26 no.1
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    • pp.40-45
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    • 2022
  • Purpose: Chronic kidney disease (CKD) has various underlying causes in children. Identification of the underlying causes of CKD is important. Genetic causes comprise a significant proportion of pediatric CKD cases. Methods: In this study, we performed whole-exome sequencing (WES) to identify genetic causes of pediatric CKD. From January to June 2021, WES was performed using samples from pediatric patients with CKD of unclear etiology. Results: Genetic causes were investigated using WES in 37 patients (17 males) with pediatric CKD stages 1 (n=5), 2 (n=7), 3 (n=2), 4 (n=2), and 5 (n=21). The underlying diseases were focal segmental glomerulosclerosis (n=9), congenital anomalies of the kidney and urinary tract including reflux nephropathy (n=8), other glomerulopathies (n=7), unknown etiology (n=6), and others (n=7). WES identified genetic causes of CKD in 12 of the 37 patients (32.4%). Genetic defects were discovered in the COL4A4 (n=2), WT1 (n=2), ACTN4, CEP290, COL4A3, CUBN, GATA3, LAMA5, NUP107, and PAX2 genes. WT1 defects were found in patients whose pathologic diagnosis was membranoproliferative glomerulonephritis, and identification of CUBN defects led to discontinuation of immunosuppressive agents. Genetic diagnosis confirmed the clinical diagnosis of hypoparathyroidism, sensorineural deafness, and renal disease; Alport syndrome; and Joubert syndrome in three of the patients with CKD of unknown etiology (COL4A4 [n=2], CUBN [n=1]). Extrarenal symptoms were considered phenotypic presentations of WT1, PAX2, and CEP290 defects. Conclusions: WES provided a genetic diagnosis that confirmed the clinical diagnosis in a significant proportion (32.4%) of patients with pediatric CKD.

Phenolic compounds from the Stems of Sapium japonicum

  • Kang, Shin-Jung;Hong, Seong-Su;Hwang, Bang-Yeon;Ro, Jai-Seup;Lee, Kyong-Soon;Towers, G.H.N.
    • Natural Product Sciences
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    • v.12 no.3
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    • pp.125-128
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    • 2006
  • A chemical examination of the stems of Sapium japonicum PAX et $H_{OFFM}$ (Euphorbiaceae) has led to the isolation of seven phenolic compounds. On the basis of UV, IR, MS, and NMR spectral data and the chemical reaction, the structures of these compounds were identified as gallic acid (1), ellagic acid (2), 3,3'-di-O-methylellagic acid (3), $4-O-({\beta}-D-xylopyranosyl)-3,3'-di-O-methylellagic$ acid (4), $4-O-({\alpha}-D-arabinofuranosyl)-3,3'-di-O-methylellagic$ acid (5), isoquercitrin (6), and geraniin (7).

Characterization of Human Thigh Adipose-derived Stem Cells (사람의 허벅지지방유래 줄기세포의 특성 분석)

  • Heo, Jin-Yeong;Yoon, Jin-Ah;Kang, Hyun-Mi;Park, Se-Ah;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.14 no.4
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    • pp.233-241
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    • 2010
  • Human adipose stem cells are an abundant, readily available population of multipotent progenitor cells that reside in adipose tissue and these cells have characteristics very similar to bone marrow mesenchymal stromal cells (BMMSCs). However, liposuction procedure, donor age, body mass index, and harvesting sites might generate differences in the initial cell population and the preparations are a heterogeneous mixture of precursors with different subsets. Therefore, in this study, we investigated the characteristics of human thigh adipose stem cells and the differentiation potential into mesodermal and endodermal lineage. Thigh adipose stem cells maintained fibroblast-like morphology similar to BM-MSCs and they underwent average 56.5 doublings and produced $5{\times}10^{22}$ cells. These cells expressed SCF, Oct4, nanog, vimentin, CK18, FGF5, NCAM, Pax6, BMP4, HNF4a, nestin, GATA4, HLA-ABC, and HLA-DR genes at p3 and they also expressed Oct4, Thy-1, FSP, vWF, vimentin, desmin, CK18, CD54, CD4, CD106, CD31, a-SMA, HLA-ABC proteins. Moreover, they could differentiate into mesodermal lineage cells such as adipocyte, osteoblast and chondrocyte. In addition, they also differentiated into insulin secreting cells in our culture condition. In conclusion, human thigh adipose stem cells retain proliferative potential and expression patterns similar to BM-MSCs and they also differentiate into various cell types. Thus, human thigh adipose stem cells might be useful alternative cell source for clinical application.

In Vitro Expansion of Homogeneous Neural Precursor Cells Derived from Human Embryonic Stem Cells

  • Na, Deuk-Chae;Kim, Se-Hee;Choi, Won-Ik;Hwang, Hyun-Jin;Han, In-Bo;Kim, Jae-Hwan;Park, Keun-Hong;Chung, Hyung-Min;Choi, Seong-Jun
    • Reproductive and Developmental Biology
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    • v.31 no.4
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    • pp.267-272
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    • 2007
  • Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo and have the capacity to differentiate into various types of cells in the body. Hence, these cells may potentially be an indefinite source of cells for cell therapy in various degenerative diseases including neuronal disorders. For clinical applications of human ES cells, directed differentiation of these cells would be necessary. The objective of this study is to develop the culture condition for the expansion of neural precursor cells derived from human ES cells. Human ES cells were able to differentiate into neural precursor cells upon a stepwise culture condition. Neural precursor cells were propagated up to 5000-fold in cell numbers over 12-week period of culture and evaluated for their characteristics. Expressions of sox1 and pax6 transcripts were dramatically up-regulated along the differentiation stages by RT-PCR analysis. In contrast, expressions of oct4 and nanog transcripts were completely disappeared in neural precursor cells. Expressions of nestin, pax6 and sox1 were also confirmed in neural precursor cells by immunocytochemical analysis. Upon differentiation, the expanded neural precursor cells differentiated into neurons, astrocytes, and oligodendrocytes. In immunocytochemical analysis, expressions of type III ${\beta}$-tubulin and MAP2ab were observed Presence of astrocytes and oligodendrocytes were also confirmed by expressions of GFAP and O4, respectively. Results of this study demonstrate the feasibility of long-term expansion of human ES cell-derived neural precursor cells in vitro, which can be a potential source of the cells for the treatment of neurodegenerative disorders.