• The Korean Journal of Mycology
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• v.46 no.4
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• pp.405-414
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• 2018

Endophytic fungi were isolated from the leaves of diverse plants inhabiting Jeju Island, Korea. The fungal isolates were identified through phylogenetic analyses incorporating nucleotide sequences derived from the internal transcribed spacer region, large subunit region of ribosomal DNA, and beta-tubulin gene. Our results identified six endophytic fungi previously unknown in Korea namely, Diaporthe goulteri, Diaporthe vaccini, Rhizosphaera pini, Valsa friesii, Xylaria primorskensis, and Zalerion arboricola were identified. Here, we present their cultural and morphological characteristics and phylogenetic relationship.

• The Korean Journal of Mycology
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• v.47 no.2
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• pp.113-120
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• 2019

We isolated endophytic fungal strains from the roots of three medicinal crops, Ligusticum chuanxiong, Angelica gigas, and Cnidium officinale, cultivated at Yeongju, Korea. The fungal strains were identified based on their morphological characteristics and molecular analyses of their internal transcribed spacer, large subunit rDNA, and beta-tubulin regions. Thereby, we identified three previously unrecorded endophytic fungal species, Dactylonectria pauciseptata, Rhizopycnis vagum, and Sistotrema sernanderi, in Korea. In this report, we describe the morphological characteristics and phylogenetic analyses of these three fungal species.

• The Korean Journal of Mycology
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• v.47 no.2
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• pp.105-112
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• 2019

A unrecorded species of Trichocladium, Trichocladium griseum, was isolated in 2017 during a survey of fungal diversity in Ulsan province, South Korea. This species was identified based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer (ITS) rDNA and ${\beta}-tubulin$ gene sequences. T. griseum has not yet been reported in South Korea. Thus, we report for the first time a new record of Trichocladium griseum in Korea, and we include the descriptions and morphological illustrations of this fungus.

• Applied Microscopy
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• v.28 no.1
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• pp.107-119
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• 1998

This study was designed to investigate the appearence and the characteristics of the apoptotic cells and the process of the joint cavity formation in mouse knee joint. Fetal mouse knee joints from 15 to 19 days of gestation were used. Paraffin-embedded serial sections, stained with H & E for light microscopic observation, Epon 812 embedded thin sections for electron microscopic observation and Lowicryl HM 20 embedded thin sections for immune-electron microscopic observation were prepared. Monoclonal antibodies to $\beta-tubulin$ and polyclonal antibodies to tissue transglutaminase were used for immune-electron microscopic study. The results obtained were as follows. 1. At 15 days of gestation, blood vessels, which have invaded in the mesenchymal cells, were present in the synovium, to form the joint cavity in the future. 2. At 16 days of gestation, the joint cleft was first appeared and several RBCs were present in the joint cleft. The invasion of blood vessels into the joint cleft was continuing, and apoptotic cells were present in the inner cell layer, adjacent to the joint cleft. Necrotic cells were also present in the outer cell layer; they were present 18 days of gestation, but apoptotic cells did not appear after 17 days of gestation. 3. In the apoptotic cells, transglutaminase were localized around vacuoles and the marginal site of the cytoplasm. 4. In the apoptotic cells, tubulin was around the endoplasmic reticulum and the marginal site of the cytoplasm. In the late stage of apoptotic cells, tubulin was localized diffusely in the cytoplasm. Tubulin was also strongly labeled around in the cytoplasm of the neighboring cell at which the apoptotic body was phagocytosed. Tubulin labeled particles were apparently increased in the seperated apoptotic bodies. On the basis of the above findings, it is proposed that during the development of the mouse knee joint, blood vessel invasion first occurs and then apoptosis and cell necrosis follow it. In the apoptotic cell, present in the synovium of the developing knee joint of the mouse. it is suggested that the redistribution of tubulin is associated with apoptotic process. And transglutaminase overexpressed in the apoptotic cell.

• Food Science and Preservation
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• v.15 no.4
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• pp.562-567
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• 2008

To develop human risk assessment protocols, we explored housekeeping gene and cytokine expression in mouse spleen cells using Rt-PCR. We normalized housekeeping gene expression by RT-PCR; gene expression was highly uniform in potato leafs and mouse spleen cells. We measured the expression of frequently used housekeeping genes, such as those encoding APRT, $\beta$-tubulin, Actin, Hsp 20.2, Cyclophilin, 18S RNA, Efla, Tbp, GAPDH, $\beta$-actin, Tuba2, Hprt, Cyclophlin A, Tfrc, and RPL13A in mice fed GM or non-GM potatoes. Housekeeping gene expression did not show any significant differences between GM and non-GM potato-fed mice. The murine model of potato-fed mice did not express IL-4 and IL-13 at a significant levels.

• Development and Reproduction
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• v.11 no.3
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• pp.235-243
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• 2007

Human umbilical cord blood(HUCB) contains a rich source of hematopoietic stem cells, mesenchymal stem cells and endothelial cell precursors. Mesenchymal stem cells(MSCs) in HUCB are multipotent stem cells, differ from hematopoietic stem cells and can be differentiated into neural cells. We studied on transdifferentiation-promoting conditions in neural cells and cholinergic neuron induction of HUCB-derived MSCs. Neural differentiation was induced by addingdimethyl sulphoxide(DMSO) and butylated hydroxyanisole(BHA) in Dulbeco's Modified Essential Medium(DMEM) and fetal bovine serum(FBS). Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor(bFGF), retinoic acid(RA) and sonic hedgehog(Shh). MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including $\beta$-tubulin III, GFAP and MBP, was markedly elevated during this acute differentiation. The differentiation rate was about $32.3{\pm}2.9%$ for $\beta$-tubulin III-positive cells, $11.0{\pm}0.9%$ for GFAP, and $9.4{\pm}1.0%$ for Gal-C. HUCB-MSCs treated combinatorially with bFGF, RA and Shh were differentiated into cholinergic neurons. After cholinergic neuronal differentiation, the $\beta$-tubulin III-positive cell population of total cells was $31.3{\pm}3.2%$ and of differentiated neuronal population, $70.0{\pm}7.8%$ was ChAT-positive showing 3 folds higher in cholinergic population than neural induction. Conclusively, HUCB-derived MSCs can be differentiated into neural and cholinergic neurons and these findings suggest that HUCB are alternative cell source of treatment for neurodegenerative diseases such as Alzheimer's disease.

• Development and Reproduction
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• v.15 no.3
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• pp.231-237
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• 2011

The objective of this study was to elucidate the dynamics of microtubules in post-ovulatory aging in vivo and in vitro of mouse oocytes. The fresh ovulated oocytes were obtained from oviducts of superovulated female ICR mice at 16 hours after hCG injection. The post-ovulatory aged oocytes were collected at 24 and 48 hours after hCG injection from in vivo and in vitro, respectively. Immunocytochemistry was performed on ${\beta}$-tubulin and acetylated ${\alpha}$-tubulin. The microtubules were localized in the spindle assembly, which was barrel-shaped or slightly pointed at its poles and located peripherally in the fresh ovulated oocytes. The frequency of misaligned metaphase chromosomes were significantly increased in post-ovulatory aged oocytes after 48 hours of hCG injection. The spindle length and width of post-ovulatory aged oocytes were significantly different from those of fresh ovulated oocytes, respectively. The staining intensity of acetylated ${\alpha}$-tubulin showed stronger in post-ovulatory aged oocytes than that in the fresh ovulated oocytes. In the aged oocytes, the spindles had moved towards the center of the oocytes from their original peripheral position and elongated, compared with the fresh ovulated oocytes. Microtubule organizing centers were formed and observed in the cytoplasm of the aged oocytes. On the contrary, it was not observed in the fresh ovulated oocytes. The alteration of spindle formation and chromosomes alignment substantiates the poor development and the increase of disorders from the post-ovulatory aged oocytes. It might be important to fertilize on time in ovulated oocytes for the developmental competence of embryos with normal karyotypes.

• BMB Reports
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• v.55 no.4
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• pp.192-197
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• 2022

Cell signals for growth factors depend on the mechanical properties of the extracellular matrix (ECM) surrounding the cells. Microtubule acetylation is involved in the transforming growth factor (TGF)-β-induced myofibroblast differentiation in the soft ECM. However, the mechanism of activation of α-tubulin acetyltransferase 1 (α-TAT1), a major α-tubulin acetyltransferase, in the soft ECM is not well defined. Here, we found that casein kinase 2 (CK2) is required for the TGF-β-induced activation of α-TAT1 that promotes microtubule acetylation in the soft matrix. Genetic mutation and pharmacological inhibition of CK2 catalytic activity specifically reduced microtubule acetylation in the cells cultured on a soft matrix rather than those cultured on a stiff matrix. Immunoprecipitation analysis showed that CK2α, a catalytic subunit of CK2, directly bound to the C-terminal domain of α-TAT1, and this interaction was more prominent in the cells cultured on the soft matrix. Moreover, the substitution of alanine with serine, the 236th amino acid located at the C-terminus, which contains the CK2-binding site of α-TAT1, significantly abrogated the TGF-β-induced microtubule acetylation in the soft matrix, indicating that the successful binding of CK2 and the C-terminus of α-TAT1 led to the phosphorylation of serine at the 236th position of amino acids in α-TAT1 and regulation of its catalytic activity. Taken together, our findings provide novel insights into the molecular mechanisms underlying the TGF-β-induced activation of α-TAT1 in a soft matrix.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8239-8245
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• 2016

The p53 gene is inactivated by the human papillomavirus (HPV) E6 protein in the majority of cervical cancers. Treatment of HeLa S3 cells with siRNA for HPV E6 permitted adenovirus-mediated transduction of a p53 gene linked to an upstream estrogen response element (ERE). Our previous study in non-siRNA treated HHUA cells, which are derived from an endometrial cancer and express estrogen receptor ${\beta}$, showed enhancing effects of an upstream ERE on adenovirus-mediated p53 gene transduction. In HeLa S3 cells treated with siRNA for HPV E6, adenovirus-mediated transduction was enhanced by an upstream ERE linked to a p53 gene carrying a proline variant at codon 72, but not for a p53 gene with arginine variant at codon 72. Expression levels of p53 mRNA and Coxsackie/adenovirus receptor (CAR) mRNA after adenovirus-mediated transfer of an ERE-linked p53 gene (proline variant at codon 72) were higher compared with those after non-ERE-linked p53 gene transfer in siRNA-treated HeLa S3 cells. Western blot analysis showed lower ${\beta}$-tubulin levels and comparatively higher p53/${\beta}$-tubulin or CAR/${\beta}$-tubulin ratios in siRNA-treated HeLa S3 cells after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with those in non-siRNA-treated cells. Apoptosis, as measured by annexin V binding, was higher after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with that after non-ERE-linked p53 gene transfer in siRNA-treated cells.

• Mycobiology
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• v.37 no.4
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• pp.251-257
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• 2009

We isolated and identified a strain of Eurotium rubrum from Meju that has not been reported in Korea. This fungus is yellowish brown; reverse dark brown on CYA and PDA while yellow on 2% MEA at $25{^{\circ}C}$. Cleistothecia are first bright yellow and gradually turned brown. Mycerial growth on CYA attained a diameter of 30 mm at $20{^{\circ}C}$, 37 mm at $25{^{\circ}C}$ and 32 mm at $30{^{\circ}C}$ after 15 days. The isolate grew slower on 2% MEA ($<$ 20 mm 15 days at $25{^{\circ}C}$) compared to CYA and PDA ($<$ 40 mm 15 days at $25{^{\circ}C}$). Cleistothecia are superficial, yellow to light brown, globose to subglobose, 40~75 ${\mu}m$ in diameter. Asci are 8-spored and globose to subglobose 8~11 ${\mu}m$. Ascospores are disciform, 4.0~5.0 ${\mu}m$ in length and 4.2~4.5 ${\mu}m$ in width. Conidia are ovate or bacillar, finely roughened to densely spinulose, 4.6~6.0 ${\mu}m$ in length and 3.0~4.3 ${\mu}m$ in width. Compared to known Eurotium rubrum, the Korean isolate showed 99% sequence similarity in ITS rDNA (554 bp) and calmodulin (750 bp) gene and 100% in $\beta$-tubulin (1016 bp) gene. The E. rubrum isolate also had weak $\beta$-glucosidase and protease activities.