• Mycobiology
• /
• v.39 no.2
• /
• pp.96-102
• /
• 2011

We investigated diet supplementation with shiitake mushroom fruiting bodies on biochemical and histological changes in hypercholesterolemic rats. Six-wk old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. A diet containing 5% Lentinus edodes fruiting bodies given to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and the LDL/high-density lipoprotein ratio by 34.33, 53.21, 75.00, 34.66, 25.73, and 71.43%, respectively. Feeding mushroom also significantly reduced body weight in hypercholesterolemic rats. However, it had no detrimental effects on plasma albumin, total bilirubin, direct bilirubin, creatinine, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that L. edodes significantly reduced plasma ${\beta}$ and pre-${\beta}$-lipoprotein but increased ${\alpha}$-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red-O staining showed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that shiitake mushrooms could be recommended as a natural cholesterol lowering substance in the diet.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.29 no.6
• /
• pp.1025-1029
• /
• 2000

This study was conducted to investigate whether a DNA comet assay could be applied for identifying irradiated pork and beef. Pork and beef were irradiated with Co-60 gamma rays at 0.1, 0.3, 0.5, 0.7 and 1.0 kGy, and stored in a freezer Cells separated from the samples were embedded in agarose gel on a slide, dissolved in a lysis solution, and electrophoresed at 2 V/cm for 2.0 min by horizontal electrophoesis. The cells were then stained with a silver staining in order to visualize the DNA using a micro-scope. The DNA fragments of the irradiated cells stretched or migrated out of the cells and formed tails towards the anode, giving the appearance of comets, while unirradiated cells formed very short or no tails. The distance of DNA migration increased with irradiation dose. Since the statistical analysis showed a significant correlation between tail length and irradiation dose, a DNA comet assay could provide not only identification but also estimation of the irradiation dose for irradiated beef and pork.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.19 no.6
• /
• pp.565-570
• /
• 1990

The role of the active oxygens on plasmid DNA damage by carbonyl compounds derived from Maillard reaction was investigated. Plasmid DNA extracted from E. coli Hb1O1 was reacted with carbonyl compounds, such as glyoxal, methyl glyoxal, dihydroxyacetone, diacetyl, glyceraldehyde, glycolaldehyde and furfural with and without the active oxygen scavengers at 37$^{\circ}C$ for 6 hours, and then the degree of damage was determined by using 1 ％ agarose gel electro-phoresis. All of the carbonyl compounds except furfural caused to damage of DNA. Among these, glyoxal, methyl glyoxal and dihydroxyacetone markedly induced the damage of DNA. On the other hand, the DNA damage by the carbonyl compounds was greatly inhibited by catalase, superoxide dismutase and $\alpha$-tocopherol it is considered that the damage of DNA is due to active oxygens, such as singlet oxygen, hydrogen peroxide and superoxide anion generated during the autoxidation of carbonyl compounds.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.29 no.5
• /
• pp.843-848
• /
• 2000

The single cell-gel electrophoresis assay (comet assay) was used to identify irradiated beans. Soy beans, kidney beans, and red beans were irradiated with $^{60}Co$ gamma rays at 0.1, 0.3, 0.5, 0.7, and 1.0 kGy. Beans were peeled out, crushed lightly, and treated with phosphate-buffered saline (PBS) to extract cells. The extracted cell suspension was mixed with agarose gel solution and spread on an agarose precoated slide. After lysis of the cells, they were subjected to microgel electrophoresis for 2 minutes, and then silver-stained. We found that the DNA fragments of the irradiated samples were stretched, migrated out of the cells, and formed tails towards the anode giving the appearance of comets, while the unirradiated or the undamaged cells formed very short or no tails. The tail lengths of irradiated samples were significantly increased as irradiation dose increased at the above 0.3 kGy.

• The Plant Pathology Journal
• /
• v.34 no.6
• /
• pp.499-505
• /
• 2018

Huanglongbing (HLB, Citrus greening disease) is one of the most devastating diseases that threaten citrus production worldwide. Although HLB presents systemically, low titer and uneven distribution of these bacteria within infected plants can make reliable detection difficult. It was known loop-mediated isothermal amplification (LAMP) method has the advantages of being highly specific, rapid, efficient, and laborsaving for detection of plant pathogens. We developed a new LAMP method targeting gene contained tandem repeat for more rapid and sensitive detection of Candidatus Liberibacter asiaticus (CLas), putative causal agent of the citrus huanglongbing. This new LAMP method was 10 folds more sensitive than conventional PCR in detecting the HLB pathogen and similar to that of real-time PCR in visual detection assay by adding SYBR Green I to mixture and 1% agarose gel electrophoresis. Positive reactions were achieved in reaction temperature 57, 60 and $62^{\circ}C$ but not $65^{\circ}C$. Although this LAMP method was not more sensitive than real-time PCR, it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Thus, we expect that this LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting the CLas in citrus and can be applied for rapid diagnosis is needed.

• Applied Chemistry for Engineering
• /
• v.33 no.2
• /
• pp.210-215
• /
• 2022

Two types of lignin materials with a different surface ionic character were used and polypyrrole layer was introduced on the lignin surface to obtain polypyrrole@lignin and polypyrrole@lignosulfonate composites using a simple chemical oxidation polymerization, reported in a previous article. Polypyrrole was effectively prepared regardless of the lignin type and the resulting composites were investigated using scanning electron microscope (SEM), cyclic voltammetry (CV), and impedance analysis. SEM and CV results showed that the obtained composites retained stable electrochemical properties after introduction of polypyrrole on the lignin surface. Impedance analyses showed that the surface properties of composites were dependent on lignin characteristics. In addition, the composites were embedded in agarose, an gelifying agent, to obtain conductive gels. It was found that the conductive gels possessed an electrical conductivity and also retained stable electrochemical properties, which indicated that the conductive gels might be useful for some applications.

• Radiation Oncology Journal
• /
• v.20 no.4
• /
• pp.367-374
• /
• 2002

Purpose : To investigate the growth inhibitory effects, and the underlying mechanism of human colon cancer cell (HT-29) death, induced by a new synthetic bile acid derivative (HS-1200). Materials and Methods : Human colon cancer cells (HT-29), in exponential growth phase, were treated with various concentrations of a new synthetic bile acid derivative (HS-1200). The growth inhibitory effects on HT-29 cells were examined using a frypan blue exclusion assay. The extent of apoptosis was determined using agarose gel electrophoresis, TUNEL assays and Hoechst staining. The apoptotic cell death was also confirmed by Western blotting of PARP, caspase-3 and DNA fragmentation factor (DFF) analysis. To investigate the involvement of mitochondria, we employed immunofluorescent staining of cytochrome c and mitochondrial membrane potential analyses. Results : The dose required for the half maximal inhibition $(IC_{50})$ of the HT-29 cell growth was $100\~150\;{\mu}M$ of HS-1200. Several changes, associated with the apoptosis of the HT-29 cells, were reveal by the agarose gel eletrophoresis, TUNEL assays and Hoechst staining, following their treatment with $100\;{\mu}M$ of HS-1200. HS-1200 treatment also induced caspase-3, PARP and DFF degradations, and the western blotting showed the processed caspase-3 p20, PARP p85 and DFF p30 and p11 cleaved products. Mitochondrial events were also demonstrated. The cytochrome c staining indicated that cytochrome c had been released from the mitochondria in the HS-1200 treated cells. The mitochondrial membrane potential $(\Delta\Psi_m)$ was also prominently decreased in the HS-1200 treated cells. Conclusion : These findings suggest that the HS-1200 - induced apoptosis of human colon cancer cells (HT-29) is mediated via caspase and mitochondrial pathways.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.6 no.6
• /
• pp.419-425
• /
• 2001

A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zironia-silica(ZS) spheres by watr-in-oil emulsification . The agarose evenly laminated the ZS bead to a depth of 30㎛, and the resultin gpellicular assembly was characterised by densities up to 2.39g/mL and a mean particle dimeter of 136 ㎛. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark ad-sorbents necessary to achieve common values of bed expansion. Furthermore, the perlicular parti-cles were characterised by improved qualities of chromatographic behaviour, particularly with re-spect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorp-tion were attributed to the physical design and pellicular deployment of the reactive surface in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks(whole fermentation broths, cell disruptates and biological extracts).

• Journal of Animal Reproduction and Biotechnology
• /
• v.38 no.4
• /
• pp.209-214
• /
• 2023

Background: Single nucleotide polymorphisms (SNPs) are widely used genetic markers with applications in human disease diagnostics, animal breeding, and evolutionary studies, but existing genotyping methods can be labor-intensive and costly. The aim of this study is to develop a simple and rapid method for identification of a single nucleotide change. Methods: A modified Polymerase Chain Reaction Amplification of Multiple Specific Alleles (PAMSA) and high resolution melt (HRM) analysis was performed to discriminate a bovine polymorphism in the NCAPG gene (rs109570900, 1326T > G). Results: The inclusion of tails in the primers enabled allele discrimination based on PCR product lengths, detected through agarose gel electrophoresis, successfully determining various genotypes, albeit with some time and labor intensity due to the use of relatively costly high-resolution agarose gels. Additionally, high-resolution melt (HRM) analysis with tailed primers effectively distinguished the GG genotype from the TT genotype in bovine muscle cell lines, offering a reliable way to distinguish SNP polymorphisms without the need for time-consuming AS-PCR. Conclusions: Our experiments demonstrated the importance of incorporating unique mismatched bases in the allele-specific primers to prevent cross-amplification by fragmented primers. This efficient and cost-effective method, as presented here, enables genotyping laboratories to analyze SNPs using standard real-time PCR.

• Microbiology and Biotechnology Letters
• /
• v.17 no.4
• /
• pp.301-306
• /
• 1989

A genomic library of Zymomonas mobilis DNA was constructed in Escherichia coli using plasmid pUC9 Allyl alcohol was used to screen a genomic clone expressing alcohol dehydrogenase. The plasmids isolated from two clones, which were sensitive to allyl alcohol, were found to be related and to share a common 2.6 kb fragment encoding alcohol dehydrogenase II identified as one of two isozymes in Z. mobilis by staining for alcohol dehydrogenase activity on polyacrylamide gel and spectrophotometric analysis of several substrate oxidations.