• The Korean Journal of Mycology
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• v.20 no.2
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• pp.118-125
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• 1992

Interspecific fusion products were obtained by protoplast fusion induced by polyethylene glycol, from auxotrophic mutants, Pleurotus cornucopiae and P. florida. The fusants were classified into allodiploidy, stable heterokaryon, and spontaneously segregated heterokaryons. Fruiting body of the clamped or clampless fusants was produced by light-dark cycle on the sawdust medium in glass bottles. Most of these clampless fusants produced mature basidiocarps. The pilei showed various mixed colors resembling the parents. All fruit bodies presented clamp connections except two fusants. When small tissues of stipe from basidiocarps were cultured on a complete medium, mycelial colonies grew more vigorously than that of the original clampless fusants. Five fusants in three crosses were analysed with the distribution of progenies and segregation of genetic characters by random spore analyses. The genetic markers were shown to segregate and recombine in the first generation of monospores isolated from basidiocarps. The analysis indicated the heter-okar-yosis and strong evidence for haploidy of vegetative nuclei, a sexual cycle consisting of nuclear fusion and meiosis. Genotypes of a large number of auxotiophic progenies were not detected. The aberration ratio of segregants indicated the gene interaction resulting from different genome structure between vegetatively incompatible species.

• Korean Journal of Food Science and Technology
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• v.32 no.3
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• pp.531-537
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• 2000

The simple microgel electrophoresis of single cells, a 'comet assay', on fruit seeds enabled the rapid identification of irradiated fruits by comparing the intact non-irradiated cells and the damaged cells of irradiated fruits. Grapes and plums were irradiated with 0.1, 0.5, 0.7, 1.0 kGy and strawberries, peaches, apples, and nectarines were irradiated with only 1.0 kGy. Seeds were isolated, crushed, and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were subjected to microgel electrophoresis for 2 minutes, and then stained. The DNA radiation-induced fragmentation of all the fruits stretched and migrated out of the cells forming a tail toward the anode giving the appearance of a comet, while the undamaged cells appeared as intact nuclei without tails. Grape and plum seeds irradiated at 0.5 kGy and higher showed significant increases in tail length. With increasing the irradiation doses, longer extention of the DNA from the nucleus toward the anode was observed. Strawberry, peach, apple, and nectarine seeds irradiated with 1.0 kGy also showed the longer tails than non-irradiated ones. DNA comet assay as a rapid and inexpensive screening technique could be an officially validated method for the detection of irradiated fruits.

• Korean Journal of Plant Resources
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• v.22 no.5
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• pp.461-466
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• 2009

This study was carried out to evaluate the effects of yam (Dioscorea batatas Dence) extracts on the cell viability, growth and nucleus-DNA damage of tobacco cells which were exposed to $\gamma$-radiation stress. The viability and growth of tobacco cells exposed to 20 Gy of radiation stress were effectively recovered by pretreatment of 10 mg/L ethylacetate (EtOAc) yam extract. Pretreatment of EtOAc extract showed 20% higher cell viability and fresh weight growth than that of cells without pretreatment in 20 Gy radiation treated tobacco cells. Nucleus-DNA damage was measured as the ratio of tail length (T) to head length (H) in individual comet image isolated from tobacco cells. The T/H ratio of control-cells and treated-cells at 20 Gy were 1.05 and 1.68, and % head DNA of those cell were 86.7 and 71.3%, respectively, suggesting that nuclei of tobacco cells were severely damaged in the integrity of DNA by the treatment of $\gamma$-radiation. However, pretreatment of MeOH, EtOAc and n-BuOH extracts decreased radiation induced DNA-damage in the tobacco cells, showing T/H ratio of 1.37, 1.01 and 1.10 and % head DNA of 81.5, 87.6 and 88.7%, respectively.

• Applied Biological Chemistry
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• v.22 no.3
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• pp.173-180
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• 1979

SDS gel electrophoresis has been employed to examine the changes in distribution of three major classes of nuclear proteins extracted from isolated liver nuclei in response to refeeding of starved rats with a fat-free high carbohydrate diet and following insulin injection into streptozotocin-diabetic rats. The relative quantity of electrophoretically separated proteins in the fraction showed marked changes with 0.14 NaCl extracts, but not with histones and phenol soluble non-histone proteins. During 48h starvation at least five proteins ranging in molecular weight from 50,000 to 180,000 daltons decreased relative to normal controls while a protein with 36,000 daltons was increased. Refeeding the starved rats with a high carbohydrate diet reversed these changes over 24 h. Insulin injection into streptozotocin-diabetic rats increased levels of the set of five 0.14 M NaCl soluble proteins identified from refeeding experiment of starved rats. The 36,000 daltons protein was also diminished. These results indicate that changes in distribution of certain nuclear proteins in 0.14M NaCl extracts are associated with the control of nuclear activity ralated to known insulin-signalled modulation and induction of cytosolic lipogenic enzymes.

• Journal of Microbiology and Biotechnology
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• v.29 no.2
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• pp.297-303
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• 2019

Corticosteroids are commonly used for pain control in rotator cuff tear. Deregulated $NF-{\kappa}B$ activation is a hallmark of chronic inflammatory diseases and has been responsible for the pathogenesis of rotator cuff tear. The Dexamethasone(DEXA) is a synthetic corticosteroid. The purpose of this study was to examine the exact effect of dexamethasone on $NF-{\kappa}B$ signaling in rotator cuff tear. We measured $NF-{\kappa}B$ expression in four groups: control, $TNF-{\alpha}$-treated, DEXA-treated, and combined treatment with $TNF-{\alpha}$ and DEXA. Tenocytes were isolated from patients with rotator cuff tears and pre-incubated with $TNF-{\alpha}$ (10 ng/ml), DEXA ($1{\mu}M$), or both of them for 10 min, 1 h, and 2 h. Expression of p65, p50, and p52 in the nuclei and cytosol was analyzed by western blotting and immunofluorescence imaging using confocal microscopy. We also evaluated nucleus/cytosol (N/C) ratios of p65, p50, and p52. In our study, the combined treatment with DEXA and $TNF-{\alpha}$ showed increased N/C ratios of p65, p50, and p52 compared with those in the $TNF-{\alpha}$ group at all time points. Additionally, in the DEXA group, N/C ratios of p65, p50, and p52 gradually increased from 10 min to 2 h. In conclusion, DEXA promoted the nuclear localization of p65, p50, and p52, but was not effective in inhibiting the inflammatory response of $TNF-{\alpha}$-stimulated rotator cuff tear.

• Development and Reproduction
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• v.10 no.1
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• pp.1-7
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• 2006

Ovarian follicular atresia in mammals is finely regulated by gonadotropins and sex steroid hormones. It is well known that granulosa cell pyknosis is a common cytological feature of atretic follicles in the ovary. The present study hypothesized that granulosa cell pyknosis during follicular atresia might be related to apoptotic process and associated with caspase-3 activation. Healthy (normal) and atretic follicles were isolated from porcine ovaries based on macro-morphological criteria. Isolated follicles were either processed for histological observation or used for collection of granulosa cells by aspiration. Hoechst 33258 staining of the cells showed a significantly higher number of fragmented nuclei, a typical morphological feature of apoptotic cell, in granulosa cells from atretic follicles than those from healthy follicles. In addition, the rate of cell death was significantly higher in granulosa cells from atretic follicles than healthy follicles, as measured by flow-cytometric cell cycle analysis. In situ detection of apoptotic cells by TUNEL revealed that apoptosis was mostly restricted to granulosa cells in follicles. Theca cells were TUNEL-negative. Finally, it has been shown by caspase-3 activity assay that granulosa cells from atretic follicles retain a higher caspase-3 activity compared to healthy follicles. Taken together, it is suggested that granulosa cell degeneration during folliclar atresia occurs by caspase-3-dependent apoptotic fashion.

• Journal of Life Science
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• v.19 no.1
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• pp.101-110
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• 2009

The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel $mas2^+$ (mitosis associated protein) gene, a homolog of human SMARCAD1 was isolated and characterized from a fission yeast Schizosaccharomyces pombe (S. pombe) using gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 922 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that an SNF2 domain is located, which is involved in the chromosome remodeling. The quantitative analysis of the $mas2^+$ transcript against $adh1^+$ showed that the expression level of $mas2^+$ is high before septum formation in S. pombe. When $mas2^+$ null mutant cells were grown at 27 and $35^{\circ}C$, the cytokinesis of $mas2^+$ null mutant was greatly delayed and a large number of multi-septate and mis-segregated cells were produced. In addition, the number of multi-septate cells significantly increased. When cells were cultured in YES rich medium to increase proliferation, the abnormal phenotypes $mas2^+$ null mutant dramatically increased. These phenotypes could be rescued by an over-expression of the mast gene. The Mas2 protein localized in the nuclei of S. pombe, as evidenced by Mas2-EGFP signals. These results suggest that the $mas2^+$ is homologous to human SMARCAD1 gene and involved in septum formation and chromosome remodeling control.

• Journal of Life Science
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• v.21 no.12
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• pp.1666-1677
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• 2011

The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel gene, the $mas1^+$($\underline{mi}$tosis $\underline{as}$sociated protein) gene, a homolog of human CIP29/Hcc1, was isolated and characterized from fission yeast Schizosaccharomyces pombe (S. pombe) using a gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 245 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that a PCB ($\underline{p}$ombe cell $\underline{c}$ycle $\underline{b}$ox) is located in the promoter region, which controls M-$G_1$ specific transcription in S. pombe. The quantitative analysis of the $mas1^+$ transcript against $adh1^+$ showed that the pattern of expression is similar to that of the septation index. Cytokinesis of mas1 mutant was greatly delayed at $25^{\circ}C$ and $36^{\circ}C$, and a large number of multi-septate cells were produced. The mas1 mutant had 2C, 4C and 6C DNA contents, as determined by FACS analysis. In addition, the number of multi-septate cells significantly increased. When cells were cultured in nitrogen starvation medium to increase proliferation, the abnormal phenotypes of mas1 mutant dramatically increased. These phenotypes could be rescued by an overexpression of the $mas1^+$ gene. The mas1 protein localized in the nuclei of S. pombe and human HeLa cells, as evidenced by Mas1-EGFP signals. The abnormal growth pattern and the morphology of mas1 mutant were complemented by a plasmid carrying human CIP29/Hcc-1cDNA. In addition, CIP29 /Hcc-1 transcript level increased in active cell proliferation stages in the developing mouse embryos. These results indicate that the $mas1^+$ ishomologous to the human CIP29/Hcc1 gene and is involved in cytokinesis and cell shape control.

• Journal of Nutrition and Health
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• v.57 no.1
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• pp.53-64
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• 2024

Purpose: Mitochondria play a crucial role in preserving skeletal muscle mass, and damage to mitochondria leads to muscle mass loss. This study investigated the effects of oxypeucedanin hydrate, a furanocoumarin isolated from Angelica dahurica radix, on myogenesis and mitochondrial function in vitro and in zebrafish models. Methods: C2C12 myotubes cultured in media containing 0.1, 1, 10, or 100 ng/mL oxypeucedanin hydrate were immunostained with myosin heavy chain (MHC), and then multinucleated MHC-positive cells were counted. The expressions of markers related to muscle differentiation, muscle protein degradation, and mitochondrial function were determined by quantitative reverse transcription polymerase chain reaction. To investigate the effects of oxypeucedanin hydrate on mitochondrial dysfunction, Tg(Xla.Eef1a1:mito-EGFP) zebrafish embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without oxypeucedanin hydrate and analyzed for mito-EGFP intensity and mitochondrial length. Results: Oxypeucedanin hydrate significantly increased MHC-positive multinucleated myotubes (≥ 3 nuclei) and increased the expression of the myogenic marker myosin heavy chain 4. However, it decreased the expressions of muscle-specific RING finger protein 1 and muscle atrophy f-box (markers of muscle protein degradation). Furthermore, oxypeucedanin hydrate enhanced the expressions of markers of mitochondrial biogenesis (peroxisome proliferator-activated receptor-gamma coactivator 1 alpha, transcription factor a mitochondrial, succinate dehydrogenase complex flavoprotein subunit A, and cytochrome c oxidase subunit 1) and mitochondrial fusion (optic atrophy 1). However, it reduced the expression of dynamin-related protein 1 (a mitochondrial fission regulator). Consistently, oxypeucedanin hydrate reduced FOLFIRI-induced mitochondrial dysfunction in the skeletal muscles of zebrafish embryos. Conclusion: The study indicates that oxypeucedanin hydrate promotes myogenesis by improving mitochondrial function, and thus, suggests oxypeucedanin hydrate has potential use as a nutritional supplement that improves muscle mass and function.

• Current Research on Agriculture and Life Sciences
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• v.1
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• pp.67-83
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• 1983

The new microsporidia S80 isolated from, Bombyx mori L. in Korea showed ovoid in the morphology of the spores and the size were measured $2.9{\pm}0.28{\mu}$ in length and $1.7{\pm}0.29{\mu}$ width. No other microsporidian spore like this has not been so far isolated from Silkworm. The length of the polar filament extruded in hydrogen peroxide ($H_2O_2$) at $30^{\circ}C$ was $26{\mu}$ of a round cytoplasm on the top. The spores were partly stained with Giemsa, Safranin-O and Gram as the same staining properties as Nosema bombycis, Microsporidia K 79 and other microsporidian spores. The fine structures were observed under scanning eleceron microscope through ultrathin sectioning. The spore wall was composed of three layers ; the thin exospore of an electron dense rippled layer, the thick electron lucent endospore which was thinning considerably at the polar filament insertion point, and the inner limiting membrane. Polar cap present at the sporeapex, with a long polar filament of 12-13 coils, subtending angle of $60^{\circ}$ to spore axis, which is tubular made up of a multilayered and are a benes core, light ring structure enclosing the dance core, the dark ring structure enclosing the inner light ring structure and the other than and light ring structure bounded from cytoplasm. Lamellate polaroplast occupied the anterior part of the spore, and the two neclei with dense nucleoplasm bounded by a double nuclear envelope were cited in the slight downer middle portion of spore. From the characteristics of the shape, size and fine structures, it is certain to reason the Microsporidia S80 belong to the phylum Microspora, class Microspora, order Microsporida, order Microsporida. The shape of two nuclei cited seems to be genus Nosema, but in the classification for the suborder it should be defined wheather pansporoblasts be formed or not and for the genis especial attempts have been made to define the characters which distinguish the disporous genera in the life cycle. Survey through the infection of the bad cocoons during 1980 to 1982 in South Korea the areas contaminated with new microsporidia were revealed 5 provinces of Kyung-Gi, Kang-Won, Chung-Nam and Chun-Nam. Pathological effects inoculated per os at second instar larvae of silkworm, the LD 50 was $7.1{\times}10^7/ml$ as lower pathogenecity than that of Nosema bombycis Naegeli of $1.2{\times}10_7/ml$. While on the other hand the inoculation of the microsporidia at fourth instar larvae lowerd the whole cocoon weight and cocoon shell weight and significant at 1% level. The microsporidia S80 defined it can not be transmitted transovarially from the result of predictive and collective examination of 21 egg batches from the infected female moth.