• Title/Summary/Keyword: heat-killed

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A Characteristics of Phagocytic Plaque on Staphylococcus aureus Layer Formed by Leukocytes of the Alcoholics (Staphylococcus aureus를 이용한 주정중독환자 백혈구의 Phagocytic Plaque 형성력)

  • 성희경;서병배;김용호
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.205-212
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    • 2002
  • This study was conducted to develop a method for direct determination of phagocytic activities in human circulatic systems and to measure the phagocytic activities in human leukocytes from the alcoholics, since phagocytic activity was considered to be very important in human immune mechanism at early stage for the health care of the alcoholics. The subjects for this study were 130 among which 95 males and 3 females were diagnosed as alcoholism and 32 was healthy blood donors. A thin layer of heat-killed Staphylococcus aureus Cowan I was placed on a plastic dish and reacted with whole blood to measure the phagocytic plaque formation by human leukocytes. In order to determine the health conditions of the subjects, some clinical laboratory tests, such as white blood cell counts, hemoglobin contents (Hgb), mean corpuscular volume of red blood cells(MCV), serum electrophoresis, B and T-lymphocytes, T-lymphocyte subtypes and phytohemagglutination test were also implemented. Compared to the non-alcoholism, new and old alcoholic inpatients showed statistically significant differences on levels of Hgb and MCV (p<0.05), but showed that T and B-lymphocyte numbers decreased and Helper T cell/Suppressor T cell ratio ($1.6{\pm}0.8$%) increased. Compared to non-alcoholism, phagocytic plaque activities of leukocytes from alcoholic patients decreased significantly and an unusual pattern in phagocytic plaque was observed, showing a strange body and chain shaped phagocytosis. Based upon these results, it is concluded that a phagocytic-plaques of Staphylococcus aureus Cowan I by leukocytes was very simple and useful method for the early immunological determination of phagocytic activities in alcoholic patients without requiring any special equipments.

Investigation of Microorganism-Based Autonomous Crack Healing Agent and Full-scale Verification of Crack Healing (미생물 기반 자발적 콘크리트 균열치유제 성능 분석 및 실스케일 균열치유성능 검증)

  • Yeon-Jun Yoo;Byung-Jae Lee;Joo-Kyoung Yang;Yun Lee
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.27 no.6
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    • pp.138-143
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    • 2023
  • In this study, the crack healing performance of each crack healing agent manufacturing method was analyzed by adding crack healing agents in the form of alginate gel and spore suspension inoculated with endospores of calcium carbonate-forming bacteria to mortar. In addition, by applying it to an full-scale structure in the form of a box-type culvert, we attempted to create an environment in which the developed crack healing agent can be applied not only to a laboratory environment but also to an actual field. The crack healing agent using the dry heat drying method showed crack healing performance, but in the case of the freeze drying method, many spores were killed by freeze hardening and therefore the crack healing performance was lost. As a result of SEM and XRD pattern analysis of the presumed crack healing material extracted from the crack of a full-scale structure, it was found to be calcite, one of the calcium carbonate crystals produced by microorganisms applied to the crack healing agent. In conclusion, it was found that the crack healing by microorganisms can be implemented in a real structure.

Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3 s.20
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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Phagocytosis of Drug-Resistant Mycobacterium Tuberculosis by Peripheral Blood Monocytes (결핵균의 약제내성과 말초혈액단핵구의 결핵균 탐식능에 관한 연구)

  • Park, Jae-Seuk;Kim, Jae-Yeal;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.470-478
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    • 1997
  • Background : Phagocytosis is probably the first step for mycobacteria to be virulent in host because virulent strains are more readily phagocytosed by macrophage than attenuated strains. According to the traditional concept, multi-drug resistant strains have been regarded as less virulent. However, this concept has been challenged, since recent studies(reported) showed that the degree of virulence and drug-resistance is not related. The purpose of this study is to evaluate whether the phagocytic activity of M.tuberculosis by peripheral blood mononuclear cells(PBMC) is different according to drug-resistance or host factor. To evaluate this, we estimated the difference of phagocytic activity of drug-resistant and drug-sensitive M.tuberculosis and also estimated the phagocytic activity of PBMC from intractable tuberculosis patients and healthy controls. Methods : PBMC from ten intractable tuberculosis patients and twelve healthy control, and three different strains of heat-killed M.tuberculosis, ie, ADS(all drug sensitive), MDR(multi-drug resistant), and ADR(all drug resistant) were used. After incubation of various strains of M.tuberculosis with PBMC, the phagocytic activity was evaluated by estimating proportion of PBMC which have phagocytosed M.tuberculosis. Results : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains(Percentage of PBMC phagocytosed M.tuberculosis in healthy control : ADS : $32.3{\pm}2.9%$, ADR : $49.6{\pm}3.4%$, p = 0.0022, Percentage of PBMC phagocytosed M.tuberculosis in intractable tuberculosis patients : ADS : $34.9{\pm}3.6%$, ADR : $50.7{\pm}4.5%$, p = 0.0069). However, there was no difference in phagocytic activity of PBMC from healthy control and intractable tuberculosis patients. Conclusion : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains and host factors does not seems to influence the phagocytosis of M.tuberculosis.

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Antibacterial and Anti-inflammatory Effects of Medicinal Plants Against Acne-inducing Bacteria (천연 약용식물 추출물의 여드름 원인균에 대한 항균 및 항염증 효과)

  • Lee, Eung-Ji;Bae, Seong-Yun;NamKung, Woo;Lee, Yong-Hwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.1
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    • pp.57-63
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    • 2010
  • To develop natural therapeutic agents for acne vulgaris, we investigated antibacterial and anti-inflammatory effects of various medicinal plant extracts. Among candidate extracts, we selected Psoralea corylifolia L. extract (AC-1) and Magnoliae officinalis extract (AC-2) which showed the relatively high antibacterial effects, and Inula helenium L. extract (ACF-1) and Chrysanthemum zawadskii var. latilobum extract (ACF-2) which showed the relatively high anti-inflammatory effects for further investigations. All of them did not show cytotoxic effects below the concentration of $50{\mu}g/mL$. The antibacterial effects of AC-1, AC-2 and extract complex (AC) against P. acnes were 2.8, 2.5 and 3.2 times higher than that of 10 % salicylic acid respectively. And the antibacterial effect of AC-2 and extract complex against S. aureus were 1.4 and 1.5 times higher than that of 10 % methylparaben respectively. Also, it was shown that ACF-1, ACF-2 and extract complex had anti-inflammatory effects. All of them exhibited inhibitory effects for the secretion of IL-8 and TNF-$\alpha$ from THP-1 cells activated by heat-killed P. acnes. They reduced about 27 %, 38 %, 44 % of IL-8 secretion and 90 %, 88 %, 90 % of TNF-$\alpha$ secretion at concentration of $50{\mu}g/mL$ respectively. These results showed that the complex of medicinal plant extracts, AC-1. AC-2, ACF-1, and ACF-2, had therapeutic effects to acne vulgaris through antibacterial and anti-inflammatory effects. Therefore, we suggest that extract complex of AC-1, AC-2, ACF-1 and ACF-2 may be used as a useful agent for development of natural cosmetics which have therapeutic effects to acne vulgaris.