• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.94-105
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• 2001

Background : Examining the biological susceptibility of Mycobacterium tuberculosis to pyrazinamide (PZA) in vitro is very difficult as PZA is inactive under normal culture conditions. The biological susceptibility test, an enzyme assay for Pzase activity, and a genetic test for pncA gene mutations, were performed in order to predict PZA resistance. Methods : 28 cultured clinical isolates of Mycobacterium tuberculosis were tested. The biological susceptibility was performed by the absolute concentration method using Lowenstein-Jensen media. The PZase activity was tested by means of Wayne's method. A 710-bp region includes the entire open reading frame of pncA was amplified and sequenced. Results : All six strains with positive PZase activity exhibited no pncA mutations with one strain showing a false resistance in the biological susceptibility test. Among the 22 strains with no PZase activity, 21 exhibited showed pncA mutations. In the biological susceptibility test, 20 strains were resistant, and one was susceptible, and the other flied to test. The mutation types varied with ten missense, one silent and one nonsense mutation 1 slipped-strand mispairing, and 6 frameshift mutations. Three strains had an adenine to guanine mutation at position -11 upstream of the start codon. Conclusion : The mutation at the pncA promotor region is frequent at -11 upstream position. Automatic sequencing of pncA is a useful tool for rapid and accurate detection of PZA resistant M. tuberculosis, and for demonstrating the epidemiological relatedness of the PZA resistant M. tuberculosis strains.

• Journal of Life Science
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• v.25 no.4
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• pp.433-440
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• 2015

Several studies suggest that regular consumption of walnuts may have beneficial effects against oxidative stress-mediated disease such as cancer. The present study reports the total phenolic and flavonoid contents, together with the antioxidant and antibacterial activities of several solvent extracts (methanol, n-hexane, ethyl acetate, n-butanol, and water) obtained from walnut (Juglans regia L.) green husk. MIC (minimal inhibitory concentration) values of the walnut extracts for 8 human pathogenic bacteria strain were determined using agar dilution method. Antioxidant activity of extracts were assessed using DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid)) assays, EC₅₀ of DPPH and ABTS scavenging activities, and determination of total phenolic and flavonoid content and its correlation with DPPH and ABTS scavenging capacities. Among the six extracts, ethyl acetate extract (EtOAc Ex) showed the highest antimicrobial activity at 3.2 mg/ml of MICs against Staphylococcus aureus SG511. Total flavonoids and polyphenol contents of EtOAc Ex were 42.48 mg of quercetin equivalents (QE)/g and 223.25 mg of gallic acid equivalents (GAE)/g respectively. The highest antioxidative potential was shown by the sample extracted with EtOAc Ex (EC₅₀=13.43 μg/ml for DPPH and EC₅₀=41.83 μg/ml for ABTS radical scavenging activity assay). These results showed that J. regia green husk extracts can be used as an easily accessible source of natural antibacterial agents and natural antioxidants.

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.210-218
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• 1990

The survival and transfer of chromosomal genes coding for the synthesis of amino acids (threonine, tryptophan, histidine, leucine, methionine) and of plasmid-borne genes coding for resistance to antibiotics (chloramphenicol, kanamycin, erythromycin) by transformation in sterile and nonsterile soil (the soil was amended to 12% vol/vol with the clay mineral, montmorillonite) was studied. In pure culture, the numbers of vegetative cells of the Bacillus subtilis strains decreased by 1 to 1.5 orders of magnitude within one week, but spores of each strain showed lesser decreases. In sterile soil, the populations of vegetative cells and spores decreased by 1.5 to 3 orders of magnitude within 2 to 4 days and then showed little additional decreased. The transformation frequencies (number of transformants/numbers of donors and recipients) of individual amino acid-genes invitro ranged from $1.3{\pm}0.6{\times}10^{-6}$ to $6.0{\pm}2.36{\times}10^{-6}$, of two amino acid-genes from $8.5{\pm}0.7{\times}10^{-8}$ to $3.1{\pm}0.6{\times}10^{-7}$, and of the antibiotic-resistance genes from $1.5{\pm} 0.2{\itmes} 10^{-7}$ to $1.4{\pm} 0.4{\times} 10^{-5}$ . In sterile soil, the frequencies of transfer of individual amino acid-genes ranged from $2.0{\times} 10^{-7}$ to $2.0{\times} 10^{-5}$ and of the antibiotic-resistance genes from $2.0{\times} 10^{-7}$ to $9.4{\pm} 4.7{\times} 10^{-6}$. The transfer of two amino acid-genes in sterile soil was detected at a frequency of $2.0{\times} 10^{-6}$ to $4.5{\times} 10^{-6}$, but only in three instances. The transformation frequencies of antibiotic-resistance genes in nonsterile soil were essentially similar to those in sterile soil. However, to detect transformants in nonsterile soil, higher concentrations of antibiotics were needed, as the result of the large numbers of indigenous soil bacteria resistant to the concentration of antibiotics used in the sterile soil and in vitro studies. The results of these studies show that genes can be transferred by transformation in soil and that this mechanism of transfer must be considered in risk assessment of the release of genetically engineered microorganisms to the environment.

• The Korean Journal of Mycology
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• v.29 no.2
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• pp.79-85
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• 2001

A strain of Dictyophora echinovolvata ASI 32002 showing good fruiting body formation was selected. Analyses of chemical and nutritional components as well as antimicrobial activity of different parts of the mushroom such as mycelium, egg, and fruiting body were carried out. There were differences in the chemical compositions and the quantities depending on developmental stages of veiled lady mushroom, D. echinovolvata ASI 32002. Nitrogen, phosphate, magnesium, and calcium in inorganic chemicals were abundant in mycelium, and potassium and mineral elements were abundant in the egg and fruiting body. Mannitol and trehalose were abundant in free sugar contents. Glutamic acid and arginine in mycelium and aspartic acid and glutamic acid in egg and fruiting body were abundant in free amino acid contents. Linoleic acid, an polyunsaturated fatty acid, was abundant in all parts of the Dictyophora species, but compositions and quantities of other fatty acids varied depending on the different parts of the mushroom. It was detected that malic acid, lactic acid and acetic acid in mycelium, formic acid, acetic acid and fumaric acid in egg, and malic acid, citric acid, lactic acid, fumaric acid in fruiting body were abundant. The methanol extracts of D. echinovolvata ASI 32002 mycelium showed antifungal activity with minimal inhibition concentration (MIC) of $62{\sim}125\;{\mu}g/ml$ that was similar levels of cyclohexamide against Aspergillus awamori, Hypocrea nigricance and Trichoderma virens. The MIC of extracts from mycelium and fruiting body against Candida albicans was $250\;{\mu}g/ml$, similar to that of tetracycline. In addition to the above results, further as food additives and ingredient of cosmetics.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2007.11a
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• pp.79-92
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• 2007

Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

• Food Science of Animal Resources
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• v.30 no.3
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• pp.435-442
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• 2010

The study analyzed the effects of salt concentration [high salt (HS) and low salt (LS)] and sodium nitrite ($NaNO_2$), which are typically utilized in Korean processing facilities, on fatty acid composition, free amino acids, microbial counts and sensory characteristics of processed dry-cured ham. Four different treatments were considered: three hams (11.30 kg) salted with 92 g/kg salt (w/w) (HS), three hams (10.65 kg) treated with HS and 100 ppm $NaNO_2$ (HS+$NaNO_2$), three hams (11.42 kg) salted with 62 g/kg salt (w/w) (LS), and three hams (10.62 kg) treated with LS and 100 ppm $NaNO_2$ (LS+$NaNO_2$). Fatty acid composition analysis revealed significantly (p<0.05) higher saturated fatty acid and lower (p<0.05) unsaturated fatty acid in the HS+$NaNO_2$ group compared with the other groups. Glutamate, alanine and lysine free amino acids were higher than the other free amino acids. The processing conditions did not significantly affect the free amino acids of biceps femoris muscles, except for the proline content (p>0.05). In sensory evaluation, the fermentation aroma of the LS group was higher than that of the HS group. The aerobic counts consistently ranged from from $2.3{\times}10^2$ to $1.11{\times}10^4$ CFU/g. Escherichia coli including strain O157:H7, Staphylococcus aureus, and Salmonella spp. were not detected.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.5
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• pp.857-863
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• 2004

The effects of adipic acid on the growth of psychrotrophic kimchi lactic acid bacteria and on the fermentatation of mulkimchi were investigated. Four Leuconostoc and one Lactobacillus species were isolated from kinnchi fermented for 50 days at 6$^{\circ}C$. The growth inhibition rate of adipic acid on psychrotrophic kimchi lactic acid bacteria and control strain, Leuconostoc mesenteroides KCCM l1324, was gradually increased from 0.1% of adipic acid concentration and its growth inhibition rate on selected strains reached 90% at 0.4% addition of adipic acid. On the bases of these results, the Preservative effect of adipic acid on the fermentation of mulkimchi was investigated for 25 days at 1$0^{\circ}C$. The pH of mulkimchi containing adipic acid was lower than that of control mulkimchi at the beginning of fermentation. However, the pH of the control mulkimchi decreased rapidly and the pH is lower at the end of fermentation than that of all samples containing adipic acid. The control increased rapidly during fermentation at the acidity. Adipic acid inhibited the growth of several microorganisms in mulkimchi including Lactobacilli. The number of Lactobacilli in control mutkimchi increased rapidly at the beginning stage of fermentation and it decreased at the end stage due to lowering of pH. However, that of mulkimchi with adipic acid slowly increased. Addition of 0.2% ethyl alcohol showed increase of preservative effect of 0.1% adipic acid in mulkimchi.

• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.132-138
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• 2019

Among several marine-derived microorganisms isolated from the coast of Jeju Island that had antimicrobial activity against fish disease pathogens, Bacillus subtilis MD-02 was tested for its dietary effect on the innate immune response and disease resistance of olive flounder. Strain MD-02 was fed to the olive flounder at a concentration of $1.2{\times}10^4$, $1.2{\times}10^6$, or $1.2{\times}10^8CFU/100g$, respectively. Consequently, the hematocrit was higher in these three groups than that in the control group at 4 weeks, and the aspartate aminotransferase and alanine aminotransferase levels were decreased in the $1.2{\times}10^8$ and $1.2{\times}10^4CFU/100$ groups compared with the control group levels. The amylase activity and total protein were significantly increased in the $1.2{\times}10^4CFU/100g$ group at 3 weeks. The innate immune response, determined from the lysozyme and macrophage activities, was higher in the $1.2{\times}10^8CFU/100g$ group than in the control group. In addition, treatment of the olive flounders with Streptococcus parauberis at $1.2{\times}10^6CFU/ml$ confirmed the mortality rate, which was 100% in the control group and 40-60% in the groups fed B. subtilis MD-02, indicating that the fish had resistance to fish disease pathogens. Therefore, it was confirmed that when fed MD-02, olive flounder builds an innate immune response and acquires resistance to fish disease pathogens, indicating that B. subtilis MD-02 can be developed as a beneficial feed additive.

• Journal of Mushroom
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• v.18 no.4
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• pp.331-338
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• 2020

This study was conducted to cultivate new Lepista nuda varieties with shorter cultivation period and better fruiting body compared to that of wild strains, for mass production and commercial application. Eighteen genetic resources of L. nuda were collected and grown in boxes using rice straw-fermented growth medium. Four lines with fruiting bodies were formed and selected as cross-breeding lines. Although 657 combinations were crossed through monospore crossing, only 17 combinations were bred between the 'CBMLN-19' line and the 'CBMLN-30' line. Among them, 8 lines with fast mycelial growth and high density were selected. After inoculating the rice straw-fermented growth medium with 14 genetic resources and 8 cross-breeding lines, their incubation period was investigated. Six of the cross-breeding lines completed their incubation in 20 days, while 7 of the 14 genetic resources took more than 40 days to complete their incubation, reducing the incubation period by more than 20 days in most cross-breeding lines. After the incubations were completed, the clay loam soil was covered with for post-cultivation, and when the mycelial cultivation was complete, the formation of fruiting bodies was induced after scraping the mycelial bodies under these environmental conditions: 14℃, 95% relative humidity or higher, and 1,500 to 2,000 ppm CO2 concentration. The temperature was reduced to 6℃ at night, resulting in a low temperature shock. Thus, 4 lines of fruiting bodies occurred from two genetic resources 'CBMLN-31' and 'CBMLN-44' and two cross-bred lines 'CBMLN-96' and 'CBMLN-103'. After inoculation, the longest period for fruiting bodies to occur was 100 days for the control:, the genetic resource 'CBMLN-31', and the shortest period (45 days) was observed for the cross-breeding line 'CBMLN-103'. The result of the investigation of the fruiting body characteristics shows that the cross-bred line 'CBMLN-103' showed a small form with 1.9 g of individual weight and 123validstipes per box, which was the highest incidence among the four lines. Another cross-bred line, 'CBMLN-96', had an individual weight of 5.5 g, which is larger than that of 'CBMLN-103'; however, the number of valid stipes per box was 30 less than that of 'CBMLN-103'. Quantity analysis showed that the control, 'CBMLN-31', had the highest quantity of 783 g per box, followed by the cross-bred line, 'CBMLN-96' with 165 g per box, and then the 'CBMLN-103' with 232 g. The quantity of the two crossbred lines was lower than that of the control 'CBMLN-31'; however, the amount of fruiting bodies was higher, and the cultivation period was shortened by 32 to 33 days. Therefore, these two lines would be selected as superior lines.

• Journal of Food Hygiene and Safety
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• v.39 no.1
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• pp.54-60
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• 2024

This study investigated the effectiveness of using pathogens and aqueous acids to reduce the Aspergillus ochraceus and Rhodotorula mucilaginosa contamination in livestock production environments. For this study, 1 mL of each bacterial suspension (10⁷-10⁸ spores/mL) was inoculated on a knife surface, dried at 37℃, and used under each treatment condition. First, to investigate the effect of organic acids, acetic, lactic, and citric acids were used. Subsequently, to select the appropriate concentration, they were prepared at concentrations of 0.5, 1, 2, 3, 4, and 5%, respectively. Accordingly, to further maximize the effect of organic acid treatment, we combined the treatment with ultraviolet light. The two strains showed a significant difference (P<0.05) compared to the initial strain, with a greater than 90% decrease in the concentrations of all organic acids. Consequently, acetic and lactic acids decreased by approximately 5 and 2 log colony forming unit (CFU)/cm², respectively, when treated with ultraviolet light (360 mJ/cm²); however, citric acid decreased by less than 1 log CFU/cm². However, when manufactured with 4% acetic acid, a severe malodor was emitted, making it difficult for workers to use it in a production environment. Accordingly, the optimal treatment conditions for organic acid and ultraviolet light for application were selected as follows: immersion in a 4% lactic acid solution for 1 minute and then, sterilization with ultraviolet light at 360 mJ/cm². Finally, when a pork meat sample was cut with a knife that was finally washed with lactic acid and treated with ultraviolet light, the low level of inoculum transferred from the cleaned knife to the surface of the sample was not detected. In conclusion, using this established method can prevent cross-contamination of the surface of the meat during processing.