Ku, Kyojin;Park, Inah;Kim, Doyeon;Kim, Jeongah;Jang, Sangwon;Choi, Mijung;Choe, Han Kyoung;Kim, Kyungjin
Molecules and Cells
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v.43
no.3
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pp.276-285
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2020
Circadian rhythm is an endogenous oscillation of about 24-h period in many physiological processes and behaviors. This daily oscillation is maintained by the molecular clock machinery with transcriptional-translational feedback loops mediated by clock genes including Period2 (Per2) and Bmal1. Recently, it was revealed that gut microbiome exerts a significant impact on the circadian physiology and behavior of its host; however, the mechanism through which it regulates the molecular clock has remained elusive. 3-(4-hydroxyphenyl)propionic acid (4-OH-PPA) and 3-phenylpropionic acid (PPA) are major metabolites exclusively produced by Clostridium sporogenes and may function as unique chemical messengers communicating with its host. In the present study, we examined if two C. sporogenes-derived metabolites can modulate the oscillation of mammalian molecular clock. Interestingly, 4-OH-PPA and PPA increased the amplitude of both PER2 and Bmal1 oscillation in a dose-dependent manner following their administration immediately after the nadir or the peak of their rhythm. The phase of PER2 oscillation responded differently depending on the mode of administration of the metabolites. In addition, using an organotypic slice culture ex vivo, treatment with 4-OH-PPA increased the amplitude and lengthened the period of PER2 oscillation in the suprachiasmatic nucleus and other tissues. In summary, two C. sporogenes-derived metabolites are involved in the regulation of circadian oscillation of Per2 and Bmal1 clock genes in the host's peripheral and central clock machineries.
To investigate symbiotic relationship of 'Chunma (Gastrodia elata) and the rhizomorph of Armillaria mellea, volatile organic acids and alcoholic compounds which were considered to be contained in Gastrodia elata were tested to determine stimulatory effects on rhizomorph growth on a chemically defined medium. Also, volatile organic acids were isolated from Gastrodia elata and analyzed by gas chromatography. The growth of rhizomorph was stimulated by the presence of alcohols and volatile organic acids, but acetic acid and methanol were ineffective. In the presence of valeric acid and ethanol, Armillaria mellea produced abundant rhizomorph at concentrations of 0.1 and 1%, respectively. Ethanol and valeric acid supplemented at regular intervals of 3 days as lower concentrations in the medium stimulated the growth of Armillaria mellea. The concentrations of ethanol and valeric acid as low at 0.01% added 3 days intervals for 15 days were more effective than initial concentrations of 0.1 and 1% in stimulating rhizomorph development of Armillaria mellea. Eight kinds of volatile organic acids were identified and quantified by gas chromatography. The major compounds were n-propionic, valeric, iso-carproic and caproic acids, and the minor compounds were iso-butyric, butyric, iso-valeric and hepatanoic acids. Valeric acid was the most abundant among them.
Since 1990, destructive yellow rots on Ganoderma lucidum caused by a soilborne fungus have been occurred in major cultivation areas of Korea. Incidences of the disease were 61% in Chulwon and 94% in Kanghwa area where the mushroom has been cultivated for 10 years, whereas the disease has not been found yet in new cultivation areas such as Moonkyung and Hongsung. when severely infected, inner tissues of bed-logs showed severe yellow and fruiting bodies of the mushroom was not produced. Infected tissues of bed-logs were readily distinguished from those of healthy ones by a distinctive brown border line. When the disease progressed, mycelia of Ganoderma lucidum were totally destroyed, and abundant ascocarps of the pathogen were formed inside the tissues of bed-logs showing yellowish green. The fungus derived from a single ascospore strongly lysed mycelia of Ganoderma lucidum growing on bottle media, and non-volatile components secreted by the pathogen were also highly inhibitory to mycelial growth of the mushroom fungus. The pathogen was identified as Arthrographis cuboidea based on its distinctive cultural and morphological characters. The fungus produced arthroconidia and unbrached conidiophores. The width of fungal conidia was distinctively wide as compared with the length. Colonies of the fungi were pale yellow to yellowish green on agar media. As a causal pathogen of yellow rot of Ganoderma lucidum., this fungus has not been reported yet in Korea.
Endogenous 84 amino acid parathyroid hormone(PTH) is synthesized as a pre-pro hormone by the chief cells of the parathyroid glands. Physiological actions of PTH include regulation of bone metabolism, renal tubular reabsorption of calcium and phosphate, and intestinal calcium absorption. In addition, PTH stimulates new bone formation by extraordinary stimulation of osteoblastic activity and decreasing calcium excretion by the kidney. In this study, we constructed and tested retrovirus vectors designed to express the human parathyroid hormone(hPTH) gene under the control of the tetracycline-inducible promoters. To increase the hPTH gene expression at turn-on state, woodchuck hepatitis virus posttranscriptional regulatory element(WPRE) sequence was also introduced into retrovirus vector at downstream region of either the hPTH gene or the sequence encoding reverse tetracycline-controlled transactivator(rtTA). Transformed primary culture cells(porcine fetal fibroblast, PFF, chicken embryonic fibroblast, CEF) were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the hPTH gene expression level using two step RT-PCR and ELISA Higher hPTH expression($3{\tims}10^4\;pg/ml,\;5.3{\times}10^4\;pg/ml$) and tighter expression control(up to 8 fold) were observed from the vector in which the WPRE sequence was placed at downstream of the hPTH gene. The resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.
Park, Sook-Young;Chi, Myoung-Hwan;Milgroom, Michael G.;Kim, Hyo-Jung;Han, Seong-Sook;Kang, Seog-Chan;Lee, Yong-Hwan
The Plant Pathology Journal
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v.26
no.4
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pp.313-320
/
2010
Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.
Yeo, Hee Dong;Jung, Ji-Young;Nam, Jung Bin;Kim, Ji Woon;Kim, Hee Kyu;Choi, Myung Suk;Alm, Glen;Rinker, Danny Lee;Yang, Jae-Kyung
Journal of the Korean Wood Science and Technology
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v.37
no.6
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pp.585-599
/
2009
This study was carried out to investigate the antifungal activity of the water soluble essential oil against Trichoderma spp. Water soluble essential oils from Pinus densiflora and Chamaecyparis obtusa were obtained from GAP (Gas assisted process) extraction apparatus. When the water soluble essential oil was treated, the growth of the conidial germination was maximum under the cultivation condition at $25^{\circ}C$ and in the culture medium adjusted to pH 5.0. The yield of water soluble essential oil was 3.9% and 3.7% in P. densiflora and C. obtusa, respectively. 24 and 15 kinds of compounds were identified in water soluble essential oils of P. densiflora and C. obtusa, respectively. The major components in the essential oil of P. densiflora were $\alpha$-Terpineol acetate and Terpinen-4-ol and those of C. obtusa were Terpinen-4-ol and $\alpha$-Terpineol. Antifungal activity was the best in the 5000 ppm of P. densiflora against Trichoderma harzianum and in 5000 ppm of C. obtusa against Trichoderma atroviride.
Lim, Jeong Cheol;Lee, Cheol Ho;Kim, Eui Ju;Choi, Byoung Ki
Journal of Wetlands Research
/
v.20
no.1
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pp.1-13
/
2018
Glaux maritima var. obtusifolia is distributed in very limited areas in South Korea and thus designated and protected as an endangered species. This study aimed to identify the diversity of vegetation in the Dangsa coast wetlands, the southern limit of G. maritima var. obtusifolia and to assess determinants of the vegetation and the importance of habitats. The phytosociological method of the $Z{\ddot{u}}rich-Montpellier$ School was used for vegetation classification and mathematical and statistical analyses were also conducted to analyze environmental factors and their relationship with the vegetation. The results of this study showed that there were 4 vegetation units in the Dangsa coast wetlands: Glaux maritima var. obtusifolia-Triglochin maritimum community (included three subcommunities), Puccinellia nipponica s.l. community, Beckmannia syzigachne-Isachne globosa community and Typha laxmannii-Phragmites communis community. It was also found that major determinants of the vegetation include moisture environment, soil depth, water level disturbance, vegetation height, community structure, etc. Glaux maritima was identified to grow most dominantly in the typicum subassociation of Glaux. maritima var. obtusifolia-Triglochin maritimum community, and the species compositions and dominant situations were observed to be similar to those in the southern limit in Japan, adjacent to South Korea. The assessment results indicated that the Dangsa coastal wetlands have a significant meaning from phytogeographical and syngeographical aspects, and contribute as a shelter for diverse species. It is required to establish conservation strategies to accurately determine the value of the wetlands of the Dangsa coast from various perspectives, and to protect and manage them.
The health benefits of garlic (Allium sativum L.) are derived from a wide variety of components and from the different ways it is administered. The known health benefits of garlic include cardiovascular protective effects, stimulation of immune function, reduction of blood glucose level, protection against microbial, viral and fungal infections, as well as anticancer effects. In the present study, it was examined the effects of water extract of A. sativum (WEAS) on the growth of cultured human tumor cells in order to investigate its anti-proliferative mechanism. Treatment of WEAS to tumor cells resulted in the growth inhibition, especially in leukemia cells, which was associated with induction of G2/M arrest of the cell cycle and apoptosis. In order to further explore the critical events leading to apoptosis in WEAS-treated U937 human leukemia cells, the following effects of WEAS on components of the mitochondrial apoptotic pathway were examined: generation of reactive oxygen species (ROS), alteration of the mitochondrial membrane potential (MMP), and the expression changes of Bcl-2 and IAP family proteins. The cytotoxic effect of WEAS was mediated by its induction of apoptosis as characterized by the occurrence of DNA ladders, apoptotic bodies and chromosome condensation in U937 cells. The WEAS-induced apoptosis in U937 cells was correlated with the generation of intracellular ROS, collapse of MMP, activation of caspase-3 and down-regulation of anti-apoptotic proteins. The quenching of ROS generation with antioxidant N-acetyl-L-cysteine conferred significant protection against WEAS-elicited ROS generation, caspase-3 activation, G2/M arrest and apoptosis. In conclusion, the present study reveals that the cellular ROS generation plays a pivotal role in the initiation of WEAS-triggered apoptotic death in U937 cells.
MicroRNAs (miRNAs) have been reported to be involved in many neurodegenerative diseases. The present study focused on the role of hsa-miR-144-3p in one of the neuro-degenerative diseases, Parkinson's disease (PD). Our study showed a remarkable down-regulation of miR-144-3p expression in 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-treated SH-SY5Y cells. MiR-144-3p was then overexpressed and silenced in human SH-SY5Y cells by miRNA-mimics and miRNA-inhibitor transfections, respectively. Furthermore, ${\beta}$-amyloid precursor protein (APP) was identified as a target gene of miR-144-3p via a luciferase reporter assay. We found that miR-144-3p overexpression significantly inhibited the protein expression of APP. Since mitochondrial dysfunction has been shown to be one of the major pathological events in PD, we also focused on the role of miR-144-3p and APP in regulating mitochondrial functions. Our study demonstrated that up-regulation of miR-144-3p increased expression of the key genes involved in maintaining mitochondrial function, including peroxisome proliferator-activated receptor ${\gamma}$ coactivator-$1{\alpha}$(PGC-$1{\alpha}$), nuclear respiratory factor 1 (NRF-1) and mitochondrial transcription factor A (TFAM). Moreover, there was also a significant increase in cellular ATP, cell viability and the relative copy number of mtDNA in the presence of miR-144-3p overexpression. In contrast, miR-144-3p silencing showed opposite effects. We also found that APP overexpression significantly decreased ATP level, cell viability, the relative copy number of mtDNA and the expression of these three genes, which reversed the effects of miR-144-3p overexpression. Taken together, these results show that miR-144-3p plays an important role in maintaining mitochondrial function, and its target gene APP is also involved in this process.
Saccharomyces cerevisiae (S. cerevisiae) and glucagon-like peptide-2 (GLP-2) have been employed to improve the intestinal development of weaned animals. The goal of this study was to determine whether either exogenous S. cerevisiae or GLP-2 elicits major effects on fecal microbiotas and cytokine responses in weaned piglets. Ninety-six piglets weaned at 26 days were assigned to one of four groups: 1) Basal diet (Control), 2) empty vector-harboring S. cerevisiae (EV-SC), 3) GLP-2-expressing S. cerevisiae (GLP2-SC), and 4) recombinant human GLP-2 (rh-GLP2). At the start of the post-weaning period (day 0), and at day 28, fecal samples were collected to assess the bacterial communities via sequencing the V1-V2 region of the 16S-rRNA gene, and piglets' blood was also sampled to measure cytokine responses (i.e., IL-$1{\beta}$, TNF-${\alpha}$, and IFN-${\gamma}$). This study revealed that, on the one hand, although S. cerevisiae supplementation did not significantly alter the growth of weaned piglets, it induced increases in the relative abundances of two core genera (Ruminococcaceae_norank and Erysipelotrichaceae_norank) and decreases in the relative abundances of two other core genera (Lachnospiraceae_norank and Clostridiale_norank) and cytokine levels (IL-$1{\beta}$ and TNF-${\alpha}$) (p < 0.05, Control vs EV-SC; p < 0.05, rh-GLP2 vs GLP2-SC). On the other hand, GLP-2 supplementation had no significant influence on fecal bacterial communities and cytokine levels, but it produced better body weight and average daily gain (p < 0.05, Control vs EV-SC; p < 0.05, rh-GLP2 vs GLP2-SC). Therefore, altered fecal microbiotas and cytokine response effects in weaned piglets were due to S. cerevisiae rather than GLP-2.
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