• The Plant Pathology Journal
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• v.37 no.6
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• pp.580-595
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• 2021

Although late blight is an important disease in ecofriendly potato cultivation in Korea, it is highly dependent on the use of eco-friendly agricultural materials and the development of biological control technology is low. It is a necessary to develop an effective biocontrol agent to inactivate late blight in the field. AFB2-2 strain is a gram-positive with peritrichous flagella. It can utilize 20 types of carbon sources, like L-arabinose, and D-trehalose at 35℃. The optimal growth temperature of the strain is 37℃. It can survive at 20-50℃ in tryptic soy broth. The maximum salt concentration tolerated by AFB2-2 strain is 7.5% NaCl. AFB2-2 strain inhibited the mycelial growth of seven plant pathogens by an average inhibitory zone of 10.2 mm or more. Among the concentrations of AFB2-2, 10⁷ cfu/ml showed the highest control value of 85.7% in the greenhouse. Among the three concentrations of AFB2-2, the disease incidence and severity of potato late blight at 10⁷ cfu/ml was lowest at 0.07 and 6.7, respectively. The nucleotide sequences of AFB2-2 strain were searched in the NCBI GenBank; Bacillus siamensis strain KCTC 13613, Bacillus velezensis strain CR-502, and Bacillus amyloliquefaciens strain DSM7 were found to have a genetic similarity of 99.7%, 99.7%, and 99.5%, respectively. The AFB2-2 strain was found to harbor the biosynthetic genes for bacillomycin D, iturin, and surfactin. Obtained data recommended that the B. velezensis AFB2-2 strain could be considered as a promising biocontrol agent for P. infestans in the field.

• Journal of Microbiology and Biotechnology
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• v.31 no.4
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• pp.592-600
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• 2021

Probiotics can be processed into a powder, tablet, or capsule form for easy intake. They are exposed to frequent stresses not only during complex processing steps, but also in the human body after intake. For this reason, various coating agents that promote probiotic bacterial stability in the intestinal environment have been developed. Silk fibroin (SF) is a material used in a variety of fields from drug delivery systems to enzyme immobilization and has potential as a coating agent for probiotics. In this study, we investigated this potential by coating probiotic strains with 0.1% or 1% water-soluble calcium (WSC), 1% SF, and 10% trehalose. Under simulated gastrointestinal conditions, cell viability, cell surface hydrophobicity, and cell adhesion to intestinal epithelial cells were then measured. The survival ratio after freeze-drying was highest upon addition of 0.1% WSC. The probiotic bacteria coated with SF showed improved survival by more than 10.0% under simulated gastric conditions and 4.8% under simulated intestinal conditions. Moreover, the cell adhesion to intestinal epithelial cells was elevated by 1.0-36.0%. Our results indicate that SF has positive effects on enhancing the survival and adhesion capacity of bacterial strains under environmental stresses, thus demonstrating its potential as a suitable coating agent to stabilize probiotics throughout processing, packaging, storage and consumption.

• Journal of Mushroom
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• v.20 no.4
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• pp.187-192
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• 2022

Wolfiporia cocos is an edible fungus commercially cultivated in Asia. To investigate metabolic changes of W. cocos mycelia under both light and dark culture conditions, gas chromatography mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) analyses were performed. In terms of the total amount of sugars, alcohols, amino acids, organic acids, fatty acids, and purines, there no significant differences between the W. cocos mycelia cultivated under light (L) or dark (D) conditions (p < 0.05). However, there were some differences with respect to the production of particular sugars and proteins. The levels of trehalose (L: 17.2 ± 0.3% vs. D: 13.9 ± 1.6%), maltose (L: 0.9 ± 0.1% vs. D: 0.3 ± 0.1%), turanose (L: 0.7 ± 0.2% vs. D: 0.1 ± 0.1%), glutamine (L: 1.6 ± 0.3% vs. D: 0.7 ± 0.2%), and proline (L: 0.3 ± 0% vs. D: 0.1 ± 0%) were all significantly higher under light condition (p < 0.05). In contrast, the levels of galactose (L: 13.7 ± 1.2% vs. D: 17.6 ± 2.0%), aspartic acid (L: 0.6 ± 0.1 % vs. D: 0.9 ± 0.1%), cystathionine (L: 0.6 ± 0.1% vs. D: 0.8 ± 0 %), and malic acid (L: 0.7 ± 0.1% vs. D: 1.2 ± 0.1%) were higher under the dark condition. It is worth noting that the amount of pachymic acid, a pharmaceutically active compound of W. cocos, was 1.68 times greater under the light condition (p < 0.05).

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.2
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• pp.130-135
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• 2022

Epididymal sperm cryopreservation provides a potential method for preserving genetic material from males of endangered species. This pilot study was conducted to develop a freezing method for tiger epididymal sperm. We evaluated post-thaw sperm condition using testes with intact epididymides obtained from a Siberian tiger (Panthera tigris altaica) after castration. The epididymis was chopped in Tyrode's albumin-lactate-pyruvate 1x and incubated at 5% CO₂, 95% air for 10 min. The Percoll separation density gradient method was used for selective recovery of motile spermatozoa after sperm collection using a cell strainer. The spermatozoa were diluted with modified Norwegian extender supplemented with 20 mM trehalose (extender 1) and subsequent extender 2 (extender 1 with 10% glycerol) and frozen using LN₂ vapor. After thawing at 37℃ for 25 s, Isolate^® solution was used for more effective recovery of live sperm. Sperm motility (computerized assisted sperm analysis, CASA), viability (SYBR-14 and Propidium Iodide) and acrosome integrity (Pisum sativum agglutinin with FITC) were evaluated. The motility of tiger epididymal spermatozoa was 40.1 ± 2.0%, and progressively motile sperm comprised 32.7 ± 2.3%. Viability was 56.3 ± 1.6% and acrosome integrity was 62.3 ± 4.4%. Cryopreservation of tiger epididymal sperm using a modified Norwegian extender and density gradient method could be effective to obtain functional spermatozoa for future assisted reproductive practices in endangered species.

• Journal of Microbiology and Biotechnology
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• v.33 no.11
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• pp.1420-1427
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• 2023

The forkhead domain genes are important for development and morphogenesis in fungi. Six forkhead genes fkhA-fkhF have been found in the genome of the model filamentous Ascomycete Aspergillus nidulans. To identify the fkh gene(s) associated with fungal development, we examined mRNA levels of these six genes and found that the level of fkhB and fkhD mRNA was significantly elevated during asexual development and in conidia. To investigate the roles of FkhB and FkhD, we generated fkhB and fkhD deletion mutants and complemented strains and investigated their phenotypes. The deletion of fkhB, but not fkhD, affected fungal growth and both sexual and asexual development. The fkhB deletion mutant exhibited decreased colony size with distinctly pigmented (reddish) asexual spores and a significantly lower number of conidia compared with these features in the wild type (WT), although the level of sterigmatocystin was unaffected by the absence of fkhB. Furthermore, the fkhB deletion mutant produced sexual fruiting bodies (cleistothecia) smaller than those of WT, implying that the fkhB gene is involved in both asexual and sexual development. In addition, fkhB deletion reduced fungal tolerance to heat stress and decreased trehalose accumulation in conidia. Overall, these results suggest that fkhB plays a key role in proper fungal growth, development, and conidial stress tolerance in A. nidulans.

• IMMUNE NETWORK
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• v.23 no.2
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• pp.16.1-16.19
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• 2023

Bacillus Calmette-Guerin (BCG) vaccine is the only licensed vaccine for tuberculosis (TB) prevention. Previously, our group demonstrated the vaccine potential of Rv0351 and Rv3628 against Mycobacterium tuberculosis (Mtb) infection by directing Th1-biased CD4⁺ T cells co-expressing IFN-γ, TNF-α, and IL-2 in the lungs. Here, we assessed immunogenicity and vaccine potential of the combined Ags (Rv0351/Rv3628) formulated in different adjuvants as subunit booster in BCG-primed mice against hypervirulent clinical Mtb strain K (Mtb K). Compared to BCG-only or subunit-only vaccine, BCG prime and subunit boost regimen exhibited significantly enhanced Th1 response. Next, we evaluated the immunogenicity to the combined Ags when formulated with four different types of monophosphoryl lipid A (MPL)-based adjuvants: 1) dimethyldioctadecylammonium bromide (DDA), MPL, and trehalose dicorynomycolate (TDM) in liposome form (DMT), 2) MPL and Poly I:C in liposome form (MP), 3) MPL, Poly I:C, and QS21 in liposome form (MPQ), and 4) MPL and Poly I:C in squalene emulsion form (MPS). MPQ and MPS displayed greater adjuvancity in Th1 induction than DMT or MP did. Especially, BCG prime and subunit-MPS boost regimen significantly reduced the bacterial loads and pulmonary inflammation against Mtb K infection when compared to BCG-only vaccine at a chronic stage of TB disease. Collectively, our findings highlighted the importance of adjuvant components and formulation to induce the enhanced protection with an optimal Th1 response.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.19-28
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• 2001

The present study was undertaken to investigate the effects of cooling rate and equilibration time on the survival, in vitro maturation and development to embryos of frozen-thawed bovine immature oocytes(Germinal Vesicle Stage). The cryoprotectants are used 10% ethylene glycol(EG) as permeating cryoprotectant and 0.05M soc.ose(S) or trehalose(T) as low molecular weight nonpermeating cryoprotectants and 5% ficoll(F) or polyvinylpyrrolidone(PVP) as high molecular weight nonpermeating cryoprotectants. Four freezing solution were uysed in this experiment(EFT: 10% EG + 5% F + 0.05M T, EFS: 10% EG + 5% F + 0.05M S, EPT: 10% EG + 5% P + 0.05M T, EPS: 10% EG + 5% P + 0.05M S). The best equilibration time and freezing solution was 15 min in EPT(83% survival rate of frozen-thawed bovine immature oocytes). When frozen-thawed bovine oocytes were cultured following IVM and IVF, there was no significant difference in cleavage and development rates among the EFT, EFS, EPT and EPS solutions. When 9 blastocysts derived from frozen bovine oocytes were transferred to 6 recipients, two recipients were pregnant. And one was aborted at 45 days of pregnancy and the other had a stillbirth.

• Journal of Mushroom
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• v.12 no.4
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• pp.316-323
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• 2014

We analyzed saccharide by dividing and comparing Monosaccharide, Disaccharide and sugar Alcohol. At first, Glucose had outstanding contained quantity of ASI 7114 with 81.11 g/l even comparing with other mushrooms for medical use and edibility. And 119.98 g/l of Fructose was observed at Hericium erinaceum that was more contained quantity than Flammulina velutipes and Lentinus edodes. But, the most contained quantity observed in Ganoderma lucidum was ASI 7015 with 15.70 g/l that was the level of 1/8 approximately against Hericium erinaceum. Ribose was found at low level generally that was hardly contained. Xylose was also observed low level. ASI 7004 was detected at 0.96g/l that was the most content with imperceptible difference by comparing with other mushrooms for medical use and edibility. Next, 35.21 g/l of Trehalose, disaccharide was observed at Agaricus bisporus that was around 11 times of content than ASI 3.09 g/l that was the most content of Ganoderma lucidum. For ${\alpha}$-Lactose, Sparassis crispa has the most amount of 3.38 g/l that was around 12.5 times of ASI 7060 0.27 g/l that was the most content of Ganoderma lucidum. For Glycerol, sugar alcohol, 64.74 g/l was observed at Pleurotus eryngii. We knew it was around 8 times of ASI 7004 8.61 g/l that was the most content of Ganoderma lucidum. 0.72 g/l of Solbitol was observed at Flammulina velutipes. We knew it was around 2times of ASI 7003 0.31 g/l that was the most content of Ganoderma lucidum. Moreover most of Ganoderma lucidum didn't contain Solbitol. 2.96 g/l of Mannitol was observed at Agaricus bisporus. that was the most content among other mushrooms. Also Mannitol was contained in Lentinus edodes and leurotus cornucopiae only. Even Ganoderma lucidum didn't have Mannitol. At last, as a result of myo-Inosito content analysis, it was seemed not to be involved in any of mushrooms.

• Food Science and Preservation
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• v.13 no.3
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• pp.375-381
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• 2006

This study was carried out to analyze the nutritional component of Lepista nuda in order to estimate its nutritional and functional values. The content of moisture were $90.90{\pm}0.09%$ carbohydrate $4.34{\pm}0.07%$, crude protein $3.70{\pm}0.03%$, ash $0.55{\pm}0.04%$, and crude fat $0.47{\pm}0.01%$, respectively The reducing sugar was $636.17{\pm}7.50mg$. Total amount of free sugar was 97.32 mg, which were mainly composed of galactose (51.98 mg) and trehalose (22.48 mg). In the results of mined analysis, the content of K was the highest as $103.10{\pm}0.18mg$, followed by $56.01{\pm}0.20mg$ of Mg. The total content of hydrolyzed amino acid was 310.39 mg which consist of essential amino acids including valine (25.40 mg), leucine (22.52 mg), lysine (21.34 mg) and 198.89 mg of non-essential amino acids including glutamic acid (60.72 mg), aspartic acid (30.17 mg), and alanine (29.72 mg), as the major amino acids. free amino acids were 220.86 mg which were composed of essential amino acids (36.21 mg) inclusive of isoleucine (21.57 mg) and threonine (11.20 mg) and non-essential amino acids (184.65 mg) including glutamic acid (42.78 mg) and aspartic acid (32.44 mg) as major amino acids. Total content of amino acid derivatives was 187.85 me and that of hydroxyproline was highest as 121.50 mg.

• Journal of Chest Surgery
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• v.30 no.12
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• pp.1159-1166
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• 1997

For Improvement of lung preservation, many tripes of preservation solution were developed and tested. The aim of this study was to compare the effect of the most frequently used extracellular type pieservation solution (Low Potassium Dextran, LPD) with a newly developed trehalose containing extracellular type preservation solution(ET-Kyoto, ETK) on postischemic lung function. Twelve New-Zealand white rabbit lungs were harvested and studied on an isolated, blood-perfused model of lung function after 4 hours of cold ischemia at $10^{\circ}C$ In group I (n=6), lungs were preserved with 100 mL/kg of LPD solution; in group II(n=6), lungs were preserved with 100 mL/kg of ETK solution. A few minutes before flushing with preservation solutions, 20$\mu\textrm{g}$ of PGEI were injected into main pulmonary artery. Functions of the preserved lung were compared with PO2, PA pressure, t acheal air pressure, and drylwet ratio. The pulmonary efferent blood oxygen tension at the end of the 60-minute reperfusion period was higher in group II compacted with group I(486.5 $\pm$ 80.3 mmHg versus $432.5\pm82.9$ mmHg at FiO2 1.0, p-value = NS). The mean pulmonary arterial pressure was similar in both $groups.(33.7\pm2.2$ mmHg versus $35.5\pm2.0$ mmHg, p-value : NS). The peak inspiratory airway pressure was significantly lower in group 11(8.010.6 mmHg versus 11.8 $\pm$ 1.4 mmHg, p-value=0.02) The water content of the lung was lower in group II $(70.2\pm6.9%$ versus 78.5 $\pm6.1%),$ but not significant. These data demonstrate that a newly-developed trehalose-containing ET-Kyoto solution yield equal or slightly superior lung function after reperfusion compared with LPD solution.