• Title/Summary/Keyword: strain recovery

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Antibacterial Activity and Probiotic Potential of Lactobacillus plantarum HKN01: A New Insight into the Morphological Changes of Antibacterial Compound-Treated Escherichia coli by Electron Microscopy

  • Sharafi, Hakimeh;Maleki, Hadi;Ahmadian, Gholamreza;Zahiri, Hossein Shahbani;Sajedinejad, Neda;Houshmand, Behzad;Vali, Hojatollah;Noghabi, Kambiz Akbari
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.225-236
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    • 2013
  • Among several bacteria examined, an antibacterial-producing Lactobacillus strain with probiotic characteristics was selected and identified based on 16S rRNA gene sequencing. Subsequent purification and mode of action of the antibacterial compounds on target cells including E. coli were investigated. Maximum production of the antibacterial compound was recorded at 18 h incubation at $30^{\circ}C$. Interestingly, antibacterial activity remained unchanged after heating at $121^{\circ}C$ for 45 min, 24 h storage in temperature range of $70^{\circ}C$ to room temperature, and 15 min exposure to UV light, and it was stable in the pH of range 2-10. The active compounds were inactivated by proteolytic enzymes, indicating their proteinaceous nature, and, therefore, referred to as bacteriocin-like inhibitory substances. Isolation and partial purification of the effective agent was done by performing ammonium sulfate precipitation and gel filtration chromatography. The molecular mass of the GFC-purified active compound (~3 kDa) was determined by Tris-Tricine SDS-PAGE. To predict the mechanisms of action, transmission electron microscopy (TEM) analysis of ultrathin sections of E. coli before and after antibacterial treatment was carried out. TEM analysis of antibacterial compounds-treated E. coli demonstrated that the completely altered bacteria appear much darker compared with the less altered bacteria, suggesting a change in the cytoplasmic composition. There were also some membrane-bound convoluted structures visible within the completely altered bacteria, which could be attributed to the response of the E. coli to the treatment with the antibacterial compound. According to the in vivo experiments oral administration of L. plantarum HKN01 resulted in recovery of infected BALB/c mice with Salmonella enterica ser. Typhimurium.

Comparison of Four Different Isolation Media for Staphylococcus aureus (황색포도상구균 분리배지 비교)

  • Oh, Min-Hee;Kang, Seong-Il;Hong, Sang-Phil;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.5
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    • pp.606-611
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    • 2009
  • Performance test was carried out between selective media which are generally used in Staphylococcus aureus isolation from food. Sensitivity, determined according to the appearance of characteristic colonies when 30 different S. aureus strains were tested, resulted as Baird-Parker agar (RPF)> $Petrifilm^{TM}$ Staph Express plate> Baird-Parker agar> Mannitol salt agar. Also, the four different media showed the same selectivity because all tested media did not produce the false positive colonies. Recovery efficiency from the artificially inoculated foodstuff was almost the same for the tested media. Presumptive colonies were collected from the dried fishery product using Mannitol salt agar and collected strains were tested on 4 different selective agar. Almost presumptive strains did not show the false positive colonies except for S. carnosus ssp carnosus. This strain was identified as false positive colonies on Mannitol salt agar, Baird-Parker agar and $Petrifilm^{TM}$ Staph Express plate. But Baird-Parker agar (RPF) did not show the false positive colonies with the same strains. So, it was concluded that the Baird-Parker agar (RPF) has more higher selectivity than other tested media in this experiment.

A Novel Acid-Stable Endo-Polygalacturonase from Penicillium oxalicum CZ1028: Purification, Characterization, and Application in the Beverage Industry

  • Cheng, Zhong;Chen, Dong;Lu, Bo;Wei, Yutuo;Xian, Liang;Li, Yi;Luo, Zhenzhen;Huang, Ribo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.989-998
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    • 2016
  • Acidic endo-polygalacturonases are the major part of pectinase preparations and extensively applied in the clarification of fruits juice, vegetables extracts, and wines. However, most of the reported fungal endo-polygalacturonases are active and stable under narrow pH range and low temperatures. In this study, an acidic endo-polygalacturonase (EPG4) was purified and characterized from a mutant strain of Penicillium oxalicum. The N-terminal amino acid sequence of EPG4 (ATTCTFSGSNGAASASKSQT) was different from those of reported endo-polygalacturonases. EPG4 displayed optimal pH and temperature at 5.0 and 60-70℃ towards polygalacturonic acid (PGA), respectively, and was notably stable at pH 2.2-7.0. When tested against pectins, EPG4 showed enzyme activity over a broad acidic pH range (>15.0% activity at pH 2.2-6.0 towards citrus pectin; and >26.6% activity at pH 2.2-7.0 towards apple pectin). The Km and Vmax values were determined as 1.27 mg/ml and 5,504.6 U/mg, respectively. The enzyme hydrolyzed PGA in endo-manner, releasing oligo-galacturonates from PGA, as determined by TLC. Addition of EPG4 (3.6 U/ml) significantly reduced the viscosity (by 42.4%) and increased the light transmittance (by 29.5%) of the papaya pulp, and increased the recovery (by 24.4%) of the papaya extraction. All of these properties make the enzyme a potential application in the beverage industry.

The Effect of 30Hz Whole Body Vibration Exercise on the Thickness and Mechanical Properties of the Erector Spinae (30Hz 전신 진동운동이 척추세움근의 두께와 기계적 속성에 미치는 영향)

  • Park, Jae-Cheol;Kim, Yong-Nam
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.1
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    • pp.45-51
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    • 2021
  • The purpose of this study was to investigate the effect of applying whole body vibration exercise at 30 Hz on the thickness and mechanical properties of the erector spinae. The study population included 24 adults (11 male and 13 female) in the N region. The subjects were divided into two groups, a 30Hz whole body vibration exercise(WG, n=12) and an LSG(12 subjects). Changes in the thickness and mechanical properties of the erector spinae were analyzed before the exercises and after four and eight weeks of exercises using a two-way repeated analysis of variance with a statistical significance level of α=0.05. When there was an interaction between the groups and measurement times, post hoc analyses were conducted at a statistical significance level of α=0.01. The thickness, frequency, stiffness and strain of the erector spinae muscles were significantly different in each time period and in the interaction between time period and the group (P<0.05), and recovery was significantly different only in each time period (P<0.05). Thus, 30Hz whole body vibration exercise had a positive effect on the thickness and mechanical properties of the erector spinae. The results of this preliminary study suggest that whole body vibration exercise may find a place as a lumbar stabilization exercise in clinical practice in the future.

Effects of Glycogen Supercompensation in Fast and Slow Muscles on Maximal Running Time of Endurance-trained Rats (단련한 흰쥐의 속근 및 지근에 Glycogen 과저장이 운동 지구력에 미치는 영향)

  • Yeoun, Dong-Soo;Hwang, Soo-Kwan;Kim, In-Sook;Ko, Seong-Kyeong;Nam, Taick-Sang;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.22 no.1
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    • pp.63-77
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    • 1988
  • This study was carried out to investigate relationships between maximal running time (MRT) and glycogen supercompensation in fast twitch white fibers (white vastus, WV), fast twitch red fibers (red vastus, RV) and slow twitch red fibers (soleus muscle, SM) of endurance-trained rats. Male rats of a Sprague-Dawley strain were divided into the trained groups and untrained groups. Untrained groups were acquired to run on the treadmill 10 minutes for 3 days and remained rest and maintained with mixed diet for 4 weeks. For last 10 days of resting period, the untrained rats were divided into 3 groups i.e. mixed diet (untrained control), high and low carbohydrate (CHO) diet groups. And each group was subdivided into 2 groups, one group was tested for the MRT and the other was sacrificed to measure the blood glucose, blood lactate, glycogen contents of liver and muscles. The experimental groups were trained on treadmill by a modified method of Constable et al. (1984) maintained with mixed diet for 4 weeks. After measurement of MRT of this group, they were also divided into high and low CHO groups and fed with these diet for 2 days and MRT of each group was measured again to see the effect of high or low CHO feeding on the MRT. Each group was maintained with the same diet for next 2 days during which some of the rats were sacrificed at given time intervals for the measurements of blood glucose and lactate, liver and the muscles glycogen. The results were summarized as follows; 1) In the untrained group, there were no significant differences between subgroups in MRT, glycogen conent of SM, RV and WV. But blood glucose concentration and glycogen content of liver of low CHO group were significantly lower than those of mixed diet group. 2) The MRT and glycogen content of SM, RV and WV of trained mixed diet group were significantly increased compared to those of untrained mixed diet group, but there was no significant difference in glycogen content of liver. 3) MRT of trained mixed, high CHO and low CHO groups were $137{\pm}9.8,\;176{\pm}9.8\;and\;129{\pm}7.3\;min$ respectively with the significant difference between them. 4) There were no differences in blood lactate concentrations between the trained high and low CHO groups immediately after maximal running and during recovery period. 5) Glycogen contents in RV and SM of trained high CHO group were significantly increased, and glycogen contents in RV, WV and liver of trained low CHO group were significantly decreased compared to those of trained mixed diet group. 6) Immediately after maximal running, the blood glucose concentrations of trained high CHO and low CHO groups were $73{\pm}4.0\;and\;67{\pm}6.9mg%$ respecitively. The blood glucose of the trained high CHO group was fully recovered within one hour by feeding. But blood glucose concentration of low CHO group was slowly recovered up to $114{\pm}4.1mg%$ after two hours of feeding and maintained. Those values were still significantly lower than that of trained mixed diet group. The synthetic rates of glycogen in liver and muscles during the recovery period followed the similar time course of the blood glucose recoveries in each group. These results suggest that an increase in MRT of trained high CHO group was attributed to the glycogen supercompensation in slow twitch muscle fibers. And a decrease in MRT of trained low CHO may be due to decreased glycogen contents of liver and muscles. The results also suggest that glycogen supercompensation was more evident in slow twitch red fibers of endurance-trained rats and blood glucose is one of the limiting factors of glycogen synthesis.

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Bioleaching of Mn(II) from Manganese Nodules by Bacillus sp. MR2 (Bacillus sp. MR2에 의한 망간단괴의 생물용출)

  • Choi, Sung-Chan;Lee, Ga-Hwa;Lee, Hong-Keum
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.411-415
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    • 2009
  • Some microorganisms are capable of leaching Mn(II) from nonsulfidic manganese ores indirectly via nonenzymatic processes. Such reductive dissolution requires organic substrates, such as glucose, sucrose, or galactose, as a source of carbon and energy for microbial growth. This study investigated characteristics of Mn(II) leaching from manganese nodules by using heterotrophic Bacillus sp. strain MR2 provided with corn starch as a less-expensive substrate. Leaching of Mn(II) at 25.6 g Mn(II) $kg^{-1}$ nodule $day^{-1}$ was accompanied with cell growth, but part of the produced Mn(II) re-adsorbed onto residual $MnO_2$ particles after 24 h. Direct contact of cells to manganese nodule was not necessary as a separation between them with a dialysis tube produced similar amount [24.6 g Mn(II) $kg^{-1}$ nodule $day^{-1}$]. These results indicated an involvement of extracellular diffusible compound(s) during Mn(II) leaching by strain MR2. In order to optimize a leaching process we tested factors that influence the reaction, and the most efficient conditions were $25\sim35^{\circ}C$, pH 5~7, inoculum density of 1.5~2.5% (v/v), pulp density of 2~3 g/L, and particle size <75 ${\mu}m$. Although Mn(II) leaching was enhanced as particle size decrease, we suggest <212 ${\mu}m$ as a proper size range since more grinding means more energy consumption The results would help for the improvement of bioleaching of manganese nodule as a less expensive, energy-efficient, and environment-friendly technology as compared to the existing physicochemical metal recovery technologies.

CELL CULTURE STUDIES OF MAREK'S DISEASE ETIOLOGICAL AGENT (조직배양(組織培養)에 의한 Marek 병(病) 병원체(病原體)의 연구(硏究))

  • Kim, Uh-Ho
    • Korean Journal of Veterinary Research
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    • v.9 no.1
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    • pp.23-62
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    • 1969
  • Throughout the studies the following experimental results were obtained and are summarized: 1. Multiplication of agents in primary cell cultures of both GF classical and CR-64 acute strain of Marek's disease infected chicken kidneys was accompanied by the formation of distinct transformed cell foci. This characteristic nature of cell transformation was passaged regularly by addition of dispersed cell from infected cultures to normal chicken kidney cell cultures, and also transferred was the nature of cell transformation to normal chick-embryo liver and neuroglial cell cultures. No cytopathic changes were noticed in inoculated chick-embryo fibroblast cultures. 2. The same cytopathic effects were noticed in normal kidney cell monolayers after the inoculation of whole blood and huffy coat cells derived from both forms of Marek's disease infected chickens. In these cases, however, the number of transformed cell foci appearing was far less than that of uninoculated monolayers prepared directly from the kidneys of Marek's disease infected chickens. 3. The change in cell culture IS regarded as a specific cell transformation focus induced by an oncogenic virus rather than it plaque in slowly progressing cytopathic effect by non-oncogenic viruses, and it is quite similar to RSV focus in chick-embryo fibroblasts in many respects. 4. The infective agent (cell transformable) were extremely cell-associated and could not be separated in an infective state from cells under the experimental conditions. 5. The focus assay of these agents was valid as shown by the high degree of linear correlation (r=0.97 and 0.99) between the relative infected cell concentration (in inoculum) and the transformed cell foci counted. 6. No differences were observed between the GF classical strain and the CR-64 acute strain of Marek's disease as far as cell culture behavior. 7. Characterization of the isolates by physical and chemical treatments, development of internuclear inclusions in Infected cells, and nucleic acid typing by differential stainings and cytochemical treatments indicated that the natures of these cell transformation agents closely resemble to those described fer the group B herpes viruses. 8. Susceptible chicks inoculated with infected kidney tissue culture cells developed specific lesions of Marek's disease, and in a case of prolonged observation after inoculation (5 weeks) the birds developed clinical symptoms and gross lesions of Marek's disease. Kidney cell cultures prepared from those inoculated birds and sacrificed showed a superior recovery of cell transformation property by formation of distinct foci. 9. Electron microscopic study of infected kidney culture cells (GF agent) by negative staining technique revealed virus particles furnishing the properties of herpes viruses. The particle was measured about $100m{\mu}$ and, so far, no herpes virus envelop has been seen from these preparations. 10. No relationship of both isolates to avian leukosis/sarcoma group viruses and PPLO was observed.

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Method of Nitrate Nitrogen Determination for Plant, Soil and Water Analysis by E. coil Cells (E. coli 세포(細胞)를 이용한 식물(植物), 토양(土壤) 및 수질(水質)의 질산태(窒酸態) 질소(窒素) 분석방법(分析方法))

  • Sohn, Sang Mok;Kucke, Martin;Lee, Yoon Gun
    • Korean Journal of Soil Science and Fertilizer
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    • v.30 no.4
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    • pp.361-369
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    • 1997
  • A microbiological nitrate determination method by E. coli is modified in Korea, using K12 wildtype, KCTC 1116, for the quantitative reduction of $NO_3{^-}$ to $NO_2{^-}$. The nitrate in plant, soil or water sample is determined spectrophotometrically after being diazotized with sulfaniamide and N-(1-naphthl)-ethlenediamine. The modified E. coli cell method and principle for nitrate determination using Korean wildtype E. coli strain is described, and cell culture and preparation of stock suspension for E. coli as well. This modified E. coli cell method can be managed simply and fast, it is suitable for the investigation of the large serials, it can be also automated and has a high degree of sensitivity up to 0.01ppm $NO_3{^-}-N$ in the sample solution. The applicability of the modified E. coli cell method has been tested for plant, soil and water analysis on a wide range of different samples. Recovery rates of added nitrate have been determined and comparisons with other standard nitrate analytical procedures have been carried out. The results with the modified E. coli cell method show high correlation ($r^2=0.98$) with those gained by the standard analytical procedures. The advantages and disadvantages of the method are also discussed to other nitrate determination methods.

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Selection and Cultural Characteristics of Whole Chicken Feather-Degrading Bacterium, Bacillus sp. SMMJ-2 (Whole Chicken Feather-Degrading Keratinolytic Protease 생산균주의 분리 및 특성)

  • Park Sung-Min;Jung Hyuck-Jun;Yu Tae-Shick
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.7-14
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    • 2006
  • Feather, generated in large quantities as a byproduct of commercial poultry processing, is almost pure keratin, which is not easily degradable by common professes. Four strains, SMMJ-2, FL-3, NO-4 and RM-12 were isolated from soil for production of extracellular keratinolytic protease. They were identified as Bacillus sp. based on their morphological and physiological characteristics. They shown high protease activity on 5.0% skim milk agar medium and produced a substrate like mucoid on keratin agar medium. Bacillus sp. SMMJ-2 had a faster production time for producing keratinolytic protease than other strains. This strain did not completely degrade whole chicken feather for five days in basal medium but completely degraded whole chicken feather when supplied with nitrogen source for 40hours in keratinolytic producing medium ($0.7%\;K_{2}HPO_{4},\;0.2%\;KH_{2}PO_{4},\;0.1%$ fructose, 1.2% whole chicken feather, $0.01%\;Na_{2}CO_3$, pH 7.0). When supplied with chicken feather as nitrogen source, keratinolytic protease activity was 89 units/ml/min. When soybean meal was used as nitrogen source, the keratinolytic protease production reached a maximum of 106 units/ml/min after 48 hours under $30^{\circ}C$, 180 agitation. To isolate the keratinolytic protease, the culture filtrate was precipitated with $(NH_4)_{2}SO_4$ and acetone. The recovery rate of keratinolytic protease was about 96% after treatment with 50% acetone. The enzyme was stable in the range of $30{\sim}50^{\circ}C$ and pH $6.0{\sim}12.0$.

The Feasibility of the DKUH-75 Left Ventricular Assist Device for Acute Cardiogenic Shock in Pigs (돼지의 급성 심인성 쇼크 모델에서 DKUH-75 좌심실보조키의 유용성에 관한 연구)

  • Park, Seong-Sik
    • Journal of Chest Surgery
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    • v.40 no.3 s.272
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    • pp.168-179
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    • 2007
  • Background: The recent trend of an increasing number of patients with acute cardiogenic shock or chronic congestive heart failure following myocardial infarction, as well as the considerable number who can not be weaned from cardiopulmonary bypass after open heart surgery, call for immediate efforts to develop affordable ventricular assist devices that are suitable for the Korean physique. Recently, a pneumatic pulsatile ventricular assist device (VAD), named DKUH-75, has been developed by the Department of Biomedical Engineering, in collaboration with the Department of Thoracic and Cardiovascular Surgery of Dankook University College of Medicine. The feasibility of the DKUH-75 VAD was evaluated on the bases of common hemodynamic variables and echocardiographic measurements in pigs, which are subjected to an acute cardiogenic shock state following myocardial infarction, using a novel coronary artery ligation method employing the ischemic preconditioning concept. Material and Method: Acute cardiogenic shock was induced in 10 Yorkshire Landrace Duroc strain pigs by ligating the left anterior descending coronary artery via an ischemic preconditioning process. The hemodynamic variables were monitored, with epicardial echocardiographic measurements performed before and one hour after the ligation. The DKUH-75 VAD was implanted into 5 pigs one hour after the onset of the shock. The hemodynamic variables and echocardiographic measurements were taken one hour after installation of the VAD. Result: The systolic, diastolic and mean systemic arterial pressures were significantly decreased in all the experimental animals one hour after the ligation. The systolic, diastolic and mean pulmonary arterial pressures were increased (Eds note: this completely contradicts the preceding statement? However, if you mean the non-experimental animals this should be stated?). The left ventricular end diastolic pressure (LVEDP) was increased, but the cardiac index decreased, An increase in the left ventricular end systolic dimension and decreases in the fractional shortening and ejection fraction were observed all animals one hour after the coronary artery ligation. In all 5 of the VAD implanted pigs, the systolic and mean systemic arterial pressures were increased, and the pulmonary arterial pressures decreased one hour after the implantation; the LVEDP decreased, but the cardiac index was significantly increased, In the echocardiographic measurements, the left ventricular end systolic dimension decreased after the implantation of the VAD, but the fractional shortening and ejection fraction significantly increased. Conclusion: Significant improvements in the hemodynamic variables and echocardiographic measurements were observed in the 5 VAD implanted animals one hour after installation, which had been subjected to an acute cardiogenic shock state by ligation of the coronary artery, indicating that the DKUH-75 VAD could help in the recovery of the myocardial function. This suggests that the DKUH-75 VAD is feasible in the short term in relation to an acute cardiogenic shock state due to myocardial infarction.