Hong, Seung-Beom;Lee, Mina;Kim, Dae-Ho;Chung, Soo-Hyun;Samson, Robert A.
한국균학회소식:학술대회논문집
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2014.05a
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pp.25-25
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2014
Koji molds are comprised of yellow, black and white. Black and white koji molds were recently re-visited by this author and it is concluded that they consists of Aspergillus luchuesnsis, A. niger and A. tubingensis, and the most important species for alcoholic beverage production is A. luchuensis. In the case of yellow koji mold, it is comprised of Aspergillus oryzae, A. sojae and A. tamari. In the case of A. sojae, the species is scarcely isolated from nature and rarely used for industry in Korea. Aspergillus tamari is often isolated from traditional Korean Meju, a fermented soybean product, and the classification of the species is clear. However, in the case of A. oryzae, differentiation between A. oryzae and A. flavus is still in controversy. In this study, we collected 415 strains of Aspergillus flavus/oryzae complex from air, rice straw, soybean, corn, peanut, arable soil and Meju in Korea and we examined the aflatoxin producing capacity of the strains. The norB-cypA, omtA and aflR genes in the aflatoxin biosynthesis gene cluster were analyzed. We found that 367 strains (88.4%) belonged to non-aflatoxigenic group (Type I of norB-cypA, IB-L-B-, IC-AO, or IA-L-B- of omtA, and AO type of aflR), and only 48 strains (11.6%) belonged to aflatoxin-producible group (Type II of norB-cypA, IC-L-B+/B- or IC-L-B+ of omtA, and AF type of aflR). In the case of A. flavus/oryzae strains from Meju, almost strains (178/192, 92.7%) belonged to non-aflatoxigenic group and only 14 strains (7.3 %) belonged to aflatoxin-producible group. It is proposed in this study that non-aflatoxigenic strain from Meju is classified as A. oryzae, considering that Meju is food material.
$TGF-{\beta}$ is one of growth factors that may be involved in the formation of bone and cartilage. Multiple studies demonstrate that $TGF-{\beta}$ is involved in regulating cell proliferation, differentiation and matrix synthesis, events observed in frature healing. The apperance of $TGF-{\beta}$ in the fracture during healing was evaluated by immunohistologic localization of $TGF-{\beta}$ using antibody. Twenty Sprauge-Dawley strain white male rats, each weiging about 150grams were used and divided two groups. The one group, the $2{\times}2mm$ bony defect was formed in the right femur. The other group, $4{\times}2mm$ bony defect was formed in right femur. Both group were sacrificed at 3day, 1, 2, 3, 4 week and femur were harvested, paraffin sections were stained with H & E, MT stain, immunihistochemical staining with $TGF-{\beta}$ antibody and observed under light microscope. The result were as follows: 1. New bone formation and cartilagenous tissue was seen at 3day. And in the $2{\times}2mm$ bony defect group, $TGF-{\beta}$ stained the cell surounding new bone. 2. The osteoclast and trabecular was seen at 1week. $TGF-{\beta}$ stained the osteoblast and in the $2{\times}2mm$ bony defect group was stained more than $4{\times}2mm$ bony defect group. 3. The lamella bone and trabecule was seen from 3, 4week, and $TGF-{\beta}$ stained almost negative. From the above finding, we could concluded that $TGF-{\beta}$ stained the osteoblast at early stage and 1week, the peak stain was seen from 1week, and then decreased, almost negative stain was seen at 3, 4week.
The compliance and stiffness of artery are closely related with disease of arteries. Pulse wave velocity(PWV) in the blood vessel is a basic and common parameter in the hemodynamics of blood pressure and blood flow wave traveling in arteries because the PWV is affected directly by the conditions of blood vessels. However, there is no standardized method to measure the PWV and it is difficult to measure. The conventional PWV measurement has being done by manual calculation of the pulse wave transmission time between coronary arterial proximal and distal points on a strip chart on which the pulse wave and ECG signal are recorded. In this study, a pressure sensor consisting of strain gauges is used to measure the blood pressure of arteries in invasive method and regular ECG electrodes are used to record the ECG signal. The R-peak point of ECG is extracted by using a reference level and time windowing technique and the ascending starting point of blood pressure is determined by using differentiation of the blood pressure signal and time windowing technique. The algorithm proposed in this study, which can measure PWV automatically, shows robust and good results in the extraction of feature points and calculation of PWV.
Human CDX2 is known as a caudal-related homeodomain transcription factor that is expressed in the intestinal epithelium and is important in differentiation and maintenance of the intestinal epithelial cells. The caudal-related homeobox proteins bind DNA according to a helix-turn-helix structure, thereby increasing the structural stability of DNA. A cancer-tumor suppressor role for Cdx2 has been shown by a decrease in the level of the expression of Cdx2 in colorectal cancer, but the mechanism of transcriptional regulation has not been examined at the molecular level. We developed a large-scale system for expression of the recombinant, novel CDX2, in Escherichia coli. A highly purified and soluble CDX2 protein was obtained in E. coli strain BL21(DE3)RIL and a hexahistidine fusion system using Ni-NTA affinity column, anion exchange, and gel filtration chromatographies. The identity and secondary structure of the purified CDX2 protein were confirmed by MALDI-TOF MS, Western blot, and a circular dichroism analyses. In addition, we studied the DNA-binding activity of recombinant CDX2 by ELISA experiment and isolated human CDX2-binding proteins derived from rat cells by an immobilized GST-fusion method. Three CDX2-binding proteins were found in the gastric tissue, and those proteins were identified to the homeobox protein Hox-D8, LIM homeobox protein 6, and SMC1L1 protein.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.26
no.4
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pp.325-336
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2000
Bone morphogenetic protein-2/4 are members of Transforming Growth Factor-$\beta$(TGF-$\beta$) superfamily and they may induce formation of cartilage and bone in vivo. This study was performed to investigate the cellular target and period of action of BMP-2/4 and understanding of actions of BMP-2/4 at cellular level. The appearance of BMP-2/4 during healing of mandibular and periodontal defect in rat was evaluated immunohistochemically. 40 Sprague-Dawley strain white male rats, each weighing about 300gm were used. Bony defect was performed in the mandible and they were sacrificed at the day of 3rd, 10th, 20th, 30th after operation. The specimens were harvested and examined histologically and immunohistochemically by localization of anti-BMP-2/4. The results were as follows: 1. Woven bone was observed at 10th day and perfect healing of defect with compact bone and periodontal ligment space at 30th day. 2. Osteoprogenitor cells, osteoblastic cells and periosteum were positive reaction to immunohistochemical stain at 10th day. 3. Cells of bone marrow space and surface cells of osteocytes and cementoblasts were positive reaction to immunohistochemical stain at 20th day. 4. Newly formed osteocytes and cementocytes were positive reaction to immunohistochemical stain at 30th day. From the above findings, we could conclude that BMP-2/4 acted significant roles as factors of induction, proliferation and differentiation during bone healing process.
Alternanthera mosaic virus (AltMV) is a member of the genus Potexvirus which has been known for less than twenty years, and has been detected in Australasia, Europe, North and South America, and Asia. The natural host range to date includes species in at least twenty-four taxonomically diverse plant families, with species in at least four other families known to be infected experimentally. AltMV has been shown to differ from Potato virus X (PVX), the type member of the genus Potexvirus, in a number of ways, including the subcellular localization of the Triple Gene Block 3 (TGB3) protein and apparent absence of interactions between TGB3 and TGB2. Differences between AltMV variants have allowed identification of viral determinants of pathogenicity, and identification of residues involved in interactions with host proteins. Infectious clones of AltMV differing significantly in symptom severity and efficiency of RNA silencing suppression have been produced, suitable either for high level protein expression (with efficient RNA silencing suppression) or for Virus-Induced Gene Silencing (VIGS; with weaker RNA silencing suppression), demonstrating a range of utility not available with most other plant viral vectors. The difference in silencing suppression efficiency was shown to be due to a single amino acid residue substitution in TGB1, and to differences in subcellular localization of TGB1 to the nucleus and nucleolus. The current state of knowledge of AltMV biology, including host range, strain differentiation, host interactions, and utility as a plant viral vector for both protein expression and VIGS are summarized.
An isolate of Peanut stunt virus (PSV) isolated from black locust tree (Robinia pseudo-acacia L.) showing severe mosaic and malformation symptoms, was designated as PSV-Rp. PSV-Rp was characterized by the tests of host range, physical properties, RNA and coat protein composition and RT-PCR analysis. Nucleotide sequences of the cucumoviruses CP genes were also used for identification and differentiation of PSV-Rp. Six plant species were used in the host range test of PSV-Rp. PSV-Rp could be differentiated from each Cucumovirus strain used as a control by symptoms of the plants. The physical properties of PSV-Rp virus were TIP $65^{\circ}C$, DEP $10^{-3}$, and LIP $2{\sim}3$ days. In dsRNA analysis, PSV-Rp consisted of four dsRNAs, but satellite RNA was not detected. Analysis of the coat proteins by SDS-PAGE showed one major protein band of about 31 kDa. RT-PCR using a part of Cucumovirus RNA3 specific primer amplified ${\sim}950bp$ DNA fragments from the crude sap of virus-infected black locust leaves. RFLP analysis of the RT-PCR product could differential PSV-RP from CMV The nucleotide sequence identity between the PSV-Rp CP and the TAV-P CP genes and the PS-V-RP CP and CMV-Y CP genes were 61.6% and 40.5%, respectively. On the other hand, the nucleotide sequence identity of the PSV-Rp CP gene was $70.9%{\sim}73.4%$ in comparison with those of PSV subgroup I (PSV-ER and PSV-J) and 67.3% with that of PSV subgroup II(PSV-W). Especially, the nucleotide sequence identity of PSV-Rp CP gene and that of PSV-Mi that was proposed recently as the type member of a novel PSV subgroup III was 92.4%.
Pleurotus eryngii has recently become a major cultivated mushroom; it uses tetrapolar heterothallism as a part of its reproductive process. Sexual development progresses only when the A and B mating types are compatible. Such mating incompatibility occasionally limits the efficiency of breeding programs in which crossing within loci-shared strains or backcrossing strategies are employed. Therefore, understanding the mating system in edible mushroom fungi will help provide a short cut in the development of new strains. We isolated and identified pheromone and receptor genes in the B3 locus of P. eryngii and performed a functional analysis of the genes in the mating process by transformation. A genomic DNA library was constructed to map the entire mating-type locus. The B3 locus was found to contain four pheromone precursor genes and four receptor genes. Remarkably, receptor PESTE3.3.1 has just 34 amino acid residues in its C-terminal cytoplasmic region; therefore, it seems likely to be a receptor-like gene. Real-time quantitative RT-PCR (real-time qRT-PCR) revealed that most pheromone and receptor genes showed significantly higher expression in monokaryotic cells than dikaryotic cells. The pheromone genes PEphb3.1 and PEphb3.3 and the receptor gene PESTE3.3.1 were transformed into P5 (A3B4). The transformants were mated with a tester strain (A4B4), and the progeny showed clamp connections and a normal fruiting body, which indicates the proposed role of these genes in mating and fruiting processes. This result also confirms that PESTE3.3.1 is a receptor gene. In this study, we identified pheromone and receptor genes in the B3 locus of P. eryngii and found that some of those genes appear to play a role in the mating and fruiting processes. These results might help elucidate the mechanism of fruiting differentiation and improve breeding efficiency.
MyoD, expressed in skeletal muscle lineages of vertebrate embryo, is one of muscle-specific basic helix-loop-helix (bHLH) transcription factors, which plays a key role in the determination and differentiation of all skeletal muscle lineages. In this study, a cDNA of grass carp MyoD was cloned and characterized from total RNA of grass carp embryos by RT-PCR. The full-length cDNA of grass carp MyoD is 1597 bp. The cDNA sequence analysis reveals an open reading frame of 825 bp coding for a protein of 275 amino acids, which includes a bHLH domain composed of basic domain (1-84th amino acids) and HLH domain (98-142th amino acids), without signal peptide. Then the MyoD cDNA of grass carp was cloned to yeast expression vector pPICZ$\alpha$A and transformed into P. pastoris GS115 strain, the recombinant MyoD protein with a molecular weight of about 31KD was obtained after inducing for 2d with 0.5% methanol in pH 8.0 BMGY medium, and the maximum yield was about 250 mg/L in shaking-flask fermentation. The results were expected to benefit for further studies on the crystal structure and physiological function of fish MyoD.
Kim, Hye-Been;Seo, Yu-Ri;Chang, Kyeong-Je;Park, Sang-Bae;Seonwoo, Hoon;Kim, Jin-Woo;Kim, Jangho;Lim, Ki-Taek
Food Engineering Progress
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v.21
no.3
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pp.233-241
/
2017
Scaffolds of cell substrates are biophysical platforms for cell attachment, proliferation, and differentiation. They ultimately play a leading-edge role in the regeneration of tissues. Recent studies have shown the potential of bioactive scaffolds (i.e., osteo-inductive) through 3D printing. In this study, rice bran-derived biocomposite was fabricated for fused deposition modeling (FDM)-based 3D printing as a potential bone-graft analogue. Rice bran by-product was blended with poly caprolactone (PCL), a synthetic commercial biodegradable polymer. An extruder with extrusion process molding was adopted to manufacture the newly blended "green material." Processing conditions affected the performance of these blends. Bio-filament composite was characterized using field emission scanning electron microscopy (FE-SEM) and energy dispersive X-ray spectroscopy (EDX). Mechanical characterization of bio-filament composite was carried out to determine stress-strain and compressive strength. Biological behaviors of bio-filament composites were also investigated by assessing cell cytotoxicity and water contact angle. EDX results of bio-filament composites indicated the presence of organic compounds. These bio-filament composites were found to have higher tensile strength than conventional PCL filament. They exhibited positive response in cytotoxicity. Biological analysis revealed better compatibility of r-PCL with rice bran. Such rice bran blended bio-filament composite was found to have higher elongation and strength compared to control PCL.
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