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Evaluation and Optimization of a Serum-based Minimum Inhibitory Concentration Assay to Caspofungin in Candida albicans Clinical Isolates

  • Yoo, Young Bin;Kim, Sung-Soon;Kim, Young Kwon;Kim, Sunghyun
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.174-183
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    • 2016
  • In the present study, a serum-based minimum inhibitory concentration (MIC) testing to caspofungin was optimized and evaluated to solve the limitations of the conventional Clinical and Laboratory Standards Institute (CLSI) guideline-based antifungal agent MIC test and the usefulness of this testing for clinical application was determined. A total of 105 Candida albicans clinical isolates were used for measuring MIC to caspofungin. Results showed that growth characteristics were different according to types of serum and the mouse serum was the most suitable for this assay. In order to measure the optimal concentration of mouse serum, 0 to 100% mouse serum were added to the media during fungal culture. The optimal concentration of serum was 50% when consideration of antifungal agent administration and inoculum size, serum components and ease of hyphae separated, and the consideration of the degree of growth. In comparison of the usefulness between the conventional Alamar-modified broth microdilution MIC assay and 50% mouse serum-based MIC testing, the range of $MIC_{80}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.42{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $2.0{\sim}32.0{\mu}g/mL$ (SD ${\pm}9.01{\mu}g/mL$). The range of $MIC_{50}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.40{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $1.0{\sim}16.0{\mu}g/mL$ (SD ${\pm}2.36{\mu}g/mL$). The MICs of 50% mouse serum-based MIC testing was increased by up to 4 to 64 times than Alamar-modified broth microdilution MIC assay. In conclusion, a 50% mouse serum-based MIC assay was more useful for measuring MIC in Candida albicans clinical isolates than conventional colorimetric broth microdilution MIC testing.

Immune Function-enhancing Effects of Bojungikkitanggami-bang (보중익기탕가미방의 면역기능 증진 효과)

  • Lee Sang Hun;Lee Seung Eon;Lee Si Hyeong;Shin Jo Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.2
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    • pp.528-533
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    • 2004
  • The immune system acts to protect the host from infectious agents that exist in the environment and from other noxious insults. The immune system has two functional divisions: the innate and the acquired. Both components involve various factors such as cytokines. A number of methodologies exist to assess aspects of immune function. There are large inter-individual variations in many immune functions even among the healthy. Genetics, age, gender, smoking habits, habitual levels of exercise, alcohol consumption, diet, stage in the female menstrual cycle, stress, history of infections and vaccinations, and early life experiences are likely to be important contributors to the observed variation. While it is clear that individuals with immune responses significantly below 'normal' are more susceptible to infectious agents and exhibit increased infectious morbidity and mortality, it is not clear how the variation in immune function among healthy individuals relates to variation in susceptibility to infection. Oriental medicine is an important factor contributing to immune competence. The author investigated the immune enhancement effects of Bojungikkitanggami-bang (BITB). The forced swimming test (FST) has been used as a screening model for new immune enhancement agents. In the present study, the author investigated the effects of BITB on FST and blood biochemical parameters related to fatigue, glucose (Glc); blood urea nitrogen (BUN); lactate dehydrogenase (LDH); creatinine; and total protein (TP). The author found that BITB (1 g/kg) significantly reduced the immobility time in the FST compared to the control. In addition, the contents of Glc, LDH, BUN, TP in the blood serum were increased in BITB (1g/kg)-fed group. Also, the author investigated the effects of BITB on the production of cytokines in human T-cell line, MOLT-4 cells. BITB (1 mg/ml) significantly increased the interferon (IFN)-vproduction compared with media control (about 2.2-fold for IFN-γ) at 24 h. However, BITB has not affect the production of IL-2 and IL-4. In addition, BITB increased the protein expression level of IFN-γ in MOLT-4 cells. Thus, BITB may have therapeutic value in generating or enhancing immune function in a clinical setting.

Channel and Data Analysis System for Digital TV Broadcasting Using Modified Hilbert Transform (변형된 힐버트 변환을 이용한 디지털 TV 방송 채널 및 데이터 분석 시스템)

  • Suh, Young-Woo;Lee, Jae-Kwon;Mok, Ha-Kyun;Choi, Jin-Yong;Seo, Jong-Soo
    • Journal of Broadcast Engineering
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    • v.14 no.4
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    • pp.438-449
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    • 2009
  • To analyze reception environments of ATSC Digital TV, CIR (Channel Impulse Response) analysis systems are widely applied. The receiving performances of conventional CIR analysis systems are not as good as those of commercial state-of-the-art receivers. There are difficulties in measuring and analyzing reception problems caused by multi-path interferences. To solve these problems, commercial DTV chip sets embedded CIR analysis system is proposed. Generally, commercial chip sets provide baseband (In-phase) channel data and field or segment sync data. For more precise analysis of measured I channel data, it is necessary to extract Q (quadrature) channel data components as well. This paper presents the technical requirements of CIR analysis system for DTV. In order to satisfy such requirements and measure more accurate magnitude and phase of CIR, a method to derive the quadrature data from the measured in-phase channel data is proposed. The proposed channel analysis system is implemented with a commercial DTV chip set and expedites the data analysis for use on DTV field test vehicles. Computer simulation and laboratory test results are provided to demonstrate the performance of the proposed channel analysis system.

Studies on the Callus Culture of Stevia as a New Sweetening Source and the Formation of Stevioside (신감미자원식물(新甘味資源植物) 스테비아의 Callus 배양(培養)과 Stevioside 생성(生成)에 관(關)한 연구(硏究))

  • Lee, Kap-Rang;Park, Jyung-Rewng;Choi, Bong-Soon;Han, Jae-Sook;Oh, Sang-Lyong;Yamada, Yasuyuki
    • Korean Journal of Food Science and Technology
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    • v.14 no.2
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    • pp.179-183
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    • 1982
  • This experiment was carried out to clarify the optimal concentrations of growth regulators for callus induction and the condition of callus culture of leaf tissue taken from Stevia rebaudiana Bertoni. The content of stevioside, sweetening component, in leaf-derived callus of stevia was also investigated. It was shown that the optimal concentrations of benzyladenine (BA) and ${\alpha}-naphthalene$acetic acid (NAA) for callus induction were $10^{-6}M$ and $10^{-5}M$, respectively. Reculture of these calli in media (Linsmaier and Skoog) supplemented with BA $10^{-6}M$ and NAA $10^{-5}M$ resulted in profuse calli 15 to 20 days after incubation. When sweetening components produced by callus were extracted and identified by TLC, stevioside appeared to have Rf value 0.50 in TLC which was exactly same as standard stevioside. Stevioside content obtained by TLC-FID analyzer was 260mg per 100g on the basis of dry weight.

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Isolation and Medium Development of the Actinomycetes, Streptomyces griseofuscus CNU-A91231, Inhibiting Phytopathogenic Fungi (식품병원성 곰팡이 성장을 저해하는 방선균 Streptomyces griseofuscus CNU-A91231의 분리 및 배지 조성)

  • Choi, Seung-Hyun;Son, Min-Jung;Kim, Sung-Han;Choi, Suk-Yul;Lee, Yoon-Hui;Choi, Jae-Eul;An, Gil-Hwan
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.322-332
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    • 2009
  • Five hundred strains of actinomycetes were screened for inhibitory activity against the phytopathogenic fungi; Alternaria alternata, Botryotinia fuckeliana, Colletotrichum acutatum, Colletotrichum gleosporioides, Corticium sasaki, Cylindrocarpon destructans, Fusarium oxysporium, Magnaporthe grisea, Phytophthora infestans, Phytium ultimum, and Thanatephorus cucumeris. The strain CNU-A91231 (Korea Agricultural Culture Collection #20938) showed a strong activity against the phytopathogenic fungi and it was identified as Streptomyces griseofuscus based on the sequence of 16s rDNA. Practical and simple media for the strain S. griseofuscus CNU-A91231 was developed at the conditions ($28^{\circ}C$ and pH 6 with aeration) for efficient bacterial growth. Alanine, glutamine, proline and ammonium ion were good nitrogen sources for the bacterium. Addition of the major salts including Na, Cl, Ca, P, K, and Mg into molasses did not increase the growth of S. griseofuscus. Addition of fertilizers containing amino acids significantly enhanced growth of the bacterium. The optimal medium was formulated as molasses + 1% of glutamate fermentation waste powder. All the conditions and components used in this study did not affect the antifungal activity of S. griseofuscus. The bacterium and the medium in this study can be used as a bio-antifungal agent for plant farming.

Morphological, Molecular, and Biochemical Characterization of Astaxanthin-Producing Green Microalga Haematococcus sp. KORDI03 (Haematococcaceae, Chlorophyta) Isolated from Korea

  • Kim, Ji Hyung;Affan, Abu;Jang, Jiyi;Kang, Mee-Hye;Ko, Ah-Ra;Jeon, Seon-Mi;Oh, Chulhong;Heo, Soo-Jin;Lee, Youn-Ho;Ju, Se-Jong;Kang, Do-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.238-246
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    • 2015
  • A unicellular red microalga was isolated from environmental freshwater in Korea, and its morphological, molecular, and biochemical properties were characterized. Morphological analysis revealed that the isolate was a unicellular biflagellated green microalga that formed a non-motile, thick-walled palmelloid or red aplanospore. To determine the taxonomical position of the isolate, its 18S rRNA and rbcL genes were sequenced and phylogenetic analysis was performed. We found that the isolate was clustered together with other related Haematococcus strains showing differences in the rbcL gene. Therefore, the isolated microalga was classified into the genus Haematococcus, and finally designated Haematococcus sp. KORDI03. The microalga could be cultivated in various culture media under a broad range of pH and temperature conditions. Compositions of the microalgal cellular components were analyzed, and its protein, carbohydrate, and lipid compositions were estimated to be 21.1 ± 0.2%, 48.8 ± 1.8%, and 22.2 ± 0.9%, respectively. In addition, D-glucose and D-mannose were the dominant monosaccharides in the isolate, and its amino acids were composed mainly of aspartic acid, glutamic acid, alanine, and leucine. Moreover, several polyunsaturated fatty acids accounted for about 80% of the total fatty acids in Haematococcus sp. KORDI03, and the astaxanthin content in the red aplanospores was estimated to be 1.8% of the dry cell weight. To the best of our knowledge, this is the first report of an Haematococcus sp. isolated from Korea, which may be used for bioresource production in the microalgal industry.

Characteristics of the inward current and its changes following fertilization in hamster eggs (햄스터 난자에서 관찰되는 내향전류의 성상과 수정후의 변화)

  • Han, Jae-hee;Hong, Seong-geun
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.280-289
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    • 1998
  • Voltage-sensitive ion channels contribute to establishment of the cell excitablity and the generation of the cellular function. At hamster oocytes in the primitive stage during developing process, an inward current elicited by voltage pulses was found to be carried mainly by $Ca^{2+}$. Even at present, $Ca^{2+}$ channels serve as the most probable route to pass this inward current but there is no evidence of the presence of this channels in eggs. To date, both the characteristic properties and the physiological role in the early stage of development remain unclear. Here we examined the characteristic properties of the inward current and changes in this currents at unfertilized oocytes, fertilized zygotes and two-cell embryos using whole-cell voltage clamp technique. The inward current carried reportedly by $Ca^{2+}$ was remained following removing external $Ca^{2+}$ but completely abolished by further replacement of impermeants such as tetramethylammonium ion ($TMA^+$) or $choline^+$ instead of $[Na^+]_0$. Tetrodotoxin did not affect on this inward current remained at $[Ca^{2+}]_0$-free condition. Removal of $Na^+$ ion out of the experimental solution clearly decreased the current. After adding 2mM $Ca^{2+}$ to the $Na^+$-free media, the inward current was restored. Interestingly, this current carried by either $Ca^{2+}$ or $Na^+$ was decreased by the reduction of intracellular $Cl^-$ concentration, or by $Cl^-$ channel blockers such as niflumic acid, DIDS and SITS. When $Cl^-$ concentration was lowered without changes in other ionic components, this inward current was reduced. At fertilized oocytes and two-cell embryos, the inward current carried by $Ca^{2+}$ and $Na^+$ was severely reduced. Also $Cl^-$ component could not be observed. From these results, the inward current is composed of $Ca^{2+}$, $Na^+$ and $Cl^-$ component, suggesting that the channel carrying this inward current is not selective specifically to $Ca^{2+}$. During early stage of development, the voltage-sensitive ion current seems not to contribute essentially to the cell cleavage and differentiation. The loss of $Cl^-$ component after fertilization suggests that $Cl^-$ may play a role in maintaining the viability of unfertilized ova.

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Influence of Calcium Supply on the Growth, Calcium and Oxalate Contents, Mineral Nutrients and Ca-oxalate Crystal Formation of Cucumber (오이생육, 칼슘, 옥살산 및 무기성분 함량 및 칼슘-옥살산염 형성에 대한 칼슘처리 효과)

  • Sung, Jwa-Kyung;Lee, Su-Yeon;Lee, Ye-Jin;Kim, Rog-Young;Lee, Ju-Young;Lee, Jong-Sik;Jang, Byoung-Choon
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.4
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    • pp.471-477
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    • 2010
  • Although the roles of calcium in plant are widely known, little is known about on an antagonistic effect of macro elements, oxalate biosynthesis and main shape of crystal in cucumber plant organs. Seeds of cucumber (Cucumis sativus cv. Ijoeunbackdadagi) were germinated in perlite tray supplied with distilled-deionized water. Seedlings were transplanted into aerated containers with a half strength of Ross nutrient solution. Ca levels treated in media were as follows; No-Ca, $Ca(NO_3)_2$ 0.25, 1.25 and 2.5 mmol $L^{-1}$, and $Ca(NO_3)_2$ 2.5 mmol $L^{-1}$ + $CaCl_210$, 25 and 50 mmol $L^{-1}$. Ca-deficient and -excessive conditions severely reduced cucumber growth, as compared to the control, and adversely affected an accumulation of macro elements (N, P, K, and Mg). Calcium favorably induced oxalate (acid-soluble) synthesis in leaves and roots of cucumber plant, but not in stem. Acid-soluble oxalate contents in leaves proportionally increased with Ca supply levels (0.91, P<0.001), however, this pattern was not observed in stem and roots. Ca-oxalate crystal formation and compositional analysis were examined using SEM-EDS technique in cucumber leaves. The main type of crystal revealed a prismatic crystal and main components were Ca, Na and Cl.

A Wavelet-Domain IKONOS Satellite Image Fusion Algorithm Considering the Spectrum Range of Multispectral Images (다중분광 영상의 색상별 스펙트럼 영역을 고려한 웨이블릿 변역 IKONOS 위성영상 융합 알고리즘)

  • Lee, Young-Gun;Kuk, Jung-Gap;Cho, Nam-Ik
    • Journal of Broadcast Engineering
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    • v.16 no.1
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    • pp.14-22
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    • 2011
  • The conventional satellite image fusion methods usually add the same amount of higher frequency components extracted from the panchromatic image to all the multispectral images. However, it is noted that each of multispectral images has different amount of overlap with the panchromatic image in terms of its spectrum, and also has different intensities. Thus giving the same amount of high frequency contents to all the spectral bands does not match with this observation, which causes color distortion in the fused image. In this paper, we propose a new wavelet-domain satellite image fusion algorithm that can compensate for these differences in intensity and spectrum overlap. For the compensation of intensity differences, we first estimate the high resolution multispectral images from P, considering the relative intensity ratios. For the compensation of the amount of spectral overlap, their wavelet coefficients are appended to the conventional wavelet-domain method where the coefficients for the addition is determined by the amount of spectrum overlap. Experiments are conducted for the IKONOS satellite images whose spectrums are well known, and the results show that the proposed algorithm gives higher PSNR and correlation coefficients compared to the conventional methods.

Effects of Gamijinhae-tang (Jiaweizhenke-tang) on Tracheal Smooth Muscle Contraction and Mucin Secretion from Airway Epithelial Cells (가미진해탕(加味鎭咳湯)이 호흡기 점액의 mucin분비에 미치는 영향)

  • Kim, Young-Ho;Joo, Ye-Jin;Jung, Hye-Mi;Seo, Un-Kyo
    • The Journal of Korean Medicine
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    • v.29 no.3
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    • pp.63-75
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    • 2008
  • Objectives: In the present study, the author intended to investigate whether Gamijinhae-tang (Jiaweizhenke-tang) (GJHT) significantly affects both contractility of tracheal smooth muscle and mucin secretion from airway epithelial cells. Materials and Methods: Effect of GJHT on contractility of isolated tracheal smooth muscle of rabbit was investigated. Confluent hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of GJHT to assess the effect of the agent on 3H-mucin secretion. At the same time, confluent NCI-H292 cells were chased for 30 min in the presence of GJHT to assess the effect of the agent on MUC5AC secretion by ELISA. Total elution profiles of control spent media and treatment sample (radioactive mucin) through Sepharose CL-4B column were analyzed. Also, effect of the agent on MUC5AC gene expression in cultured NCI-H292 cells was investigated. Possible cytotoxicities of the agent were assessed by measuring both lactate dehydrogenase (LDH) release from HTSE cells and examining the rate of survival and proliferation of NCI-H292 cells. Results: (1) GJHT inhibited Ach-induced contraction of isolated tracheal smooth muscle; (2) GJHT significantly increased mucin secretion from cultured HTSE cells. However, it did not affect MUC5AC secretion from NCI-H292 cells, only chiefly affecting the 'mucin' secretion; (3) GJHT did not significantly affect the expression levels of MUC5AC gene in cultured NCI-H292 cells; (4) GJHT did not significantly inhibit the survival and proliferation of NCI-H292 cells. However, it slightly increased LDH release from HTSE cells. Conclusion: The author suggests that effects of GJHT with their components should be further investigated and it is valuable to find, from oriental medical prescriptions, novel agents which might regulate mucin secretion from airway epithelial cells.

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