• Title/Summary/Keyword: Diluted factor

Search Result 60, Processing Time 0.023 seconds

Comparison of Quantitative Endotoxin against 5 Species of Enterobacteriaceae (장내세균 5종의 Endotoxin 정량 비교)

  • Kwon, Pil Seung
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.48 no.2
    • /
    • pp.124-129
    • /
    • 2016
  • Endotoxin, also known as lipopolysaccharide (LPS) produced by the cell wall of gram negative bacteria can be present in any liquid or on any biomaterial. Endotoxin in blood can cause fever and inflammation. In this study, we compared bacterial endotoxin using Escherichia coli O157:H7, Klebsiella oxytoca, Salmonella Typhi, Shigella sonnei and Morganella morganii. Bacteria were cultured for use in the experiment, and diluted to $1.5{\times}10^8CFU/mL$. A check marked sensitivity confirmatory test of the Limulus amebocyte lysate (LAL) reagent was performed to examine the validity. The end point reaction to each bacteria sample was confirmed with 10 fold dilution and then the final reaction end point was confirmed by 2 fold dilution between the dilution step and the upper dilution step. According to the results, in detection of endotoxins in more than 0.015 EU/mL, E. coli O157 was 75~37.5 CFU/mL, K. oxytoca 37.5~18.75 CFU/mL, M. morganii and S. Typhi 3.75~1.875 CFU/mL, and S. sonnei 7.5~3.75 CFU/mL. The resulting value was finally ensured by a confirmation test for the inhibitory factor. Based on this study, conduct of further research on bacterial endotoxin is encouraged.

Effect of Bordeaux Mixture on Control of Rice Leaf Blast (벼 유기재배에서 석회보르도액을 이용한 벼 잎도열병 방제 효과)

  • Kang, Beom-Ryong;Kim, Seon-Gon;Kim, Do-Ik;Lee, Yong-Hwan;Choi, Kyong-Ju;Choi, Yong-Soo
    • Research in Plant Disease
    • /
    • v.14 no.3
    • /
    • pp.182-186
    • /
    • 2008
  • Recently organic farming practice of rice has been emerged in Korea, but one of the major limiting factor is the no effective environmental-friendly agro-materials to control major plant diseases. Bordeaux mixture has been used effectively as a preventive agro-chemical. The aim of this study was to investigate efficacy of Bordeanux mixture on control of rice blast caused by Magnaporthe grisea which is one of the disruptive rice diseases in world-wide. In greenhouse experiment, pre-treatment of 6-6 type of Bordeaux mixture before inoculation of spore suspension of M. grasea showed 71 % of control value. In field experiment, preventive applications of 4-8 and 6-6 types of Bordeaux mixture showed over 71 % of the control value. Chemical injury on rice leaves were not found in the application concentrations of all types of Bordeaux mixture, but observed in applications of Bordeaux mixtures between 30 and 100 diluted concentrations. This results indicate Bordeaux mixture can be used as an effective environmental-friendly agro-chemical to control rice blast disease in the field.

Effects of Hominis Placenta Pharmacopucture in a Chronic Physical Stress Rat Model on Changes in Behavior, Weight, Sleep-Related Hormones, Inflammatory Cytokines, and Anti-Oxidant Enzymes (자하거 약침이 만성 신체 스트레스 모델 백서의 행동, 체중, 수면 호르몬, 염증 사이토카인, 항산화 효소의 변화에 미치는 영향)

  • Ha, Ji-Won;Lee, Yu-Mi;Na, Chang-Su;Sakong, Jong-Won;Kim, Bo-Kyung
    • Journal of Oriental Neuropsychiatry
    • /
    • v.30 no.4
    • /
    • pp.357-369
    • /
    • 2019
  • Objectives: This study was designed to examine the effect of Hominis Placenta Pharmacopuncture on the change in behavior, sleep-related hormones, inflammatory cytokines, anti-oxidants, weight, blood, and serum on rats given chronic physical stress. Methods: Wistar rats older than age 10 weeks were used in this experiment. They were divided into six groups. The normal group was not given stress. The control group was given only chronic physical stress and no treatment. The positive control group was given chronic physical stress and treated with zolpidem. Three Hominis Placenta Pharmacopuncture (HPP) groups were given chronic physical stress, then treated with different concentrations of HPP; HPP-0.5× (0.5 times diluted), HPP-1× (undiluted) and HPP-2× (2 times condensed). The changes of values of Nestlet Shredding results, weight, Melatonin, Gamma-aminobutylic Acid (GABA), Tumor Necrosis Factor Alpha (TNF-α), Interleukin-6 (IL-6), Superoxide dismutase (SOD), glutathione peroxidase (GPx), AST, ALT, BUN, Creatinine, and leukocytes were observed during the experiment. Results: The changes in chronic physical stress of the rat model were as follow. The Nestlet Shredding result increased in the control group compared to the normal group (p<0.05), and decreased in the HPP-1× and HPP-2× compared to the control group (p<0.05). The amount of weight gain showed increasing tendency in the HPP-2× compared to the control group since the second week. The GABA increased (p<0.05) and Melatonin also showed certain increasing tendency in the HPP-1× and HPP-2× compared to the control group. The TNF-α and IL-6 increased in the control group compared to the normal group (p<0.01), and decreased in all the HPPs compared to the control group (p<0.05). The SOD level decreased in the control group compared to the normal group (p<0.01), and increased in all the HPPs compared to the control group (p<0.05). GPx, AST, ALT, Bun, Creatinine and leukocytes showed no noticeable difference among all groups. Conclusions: Hominis Placenta Pharmacopuncture was effective in increasing weight, GABA, Melatonin, SOD, and decreasing Nestlet Shredding and inflammatory cytokines, suggesting that it consequently facilitates in relieving physical stress.

The Buffer Capacity of the Carbonate System in the Southern Korean Surface Waters in Summer (하계 한국 남부해역 표층수의 탄산계 완충역량)

  • HWANG, YOUNGBEEN;LEE, TONGSUP
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
    • /
    • v.27 no.1
    • /
    • pp.17-32
    • /
    • 2022
  • The buffer capacity of southern Korean waters in summer was quantified using data set of temperature, salinity, dissolved inorganic carbon, total alkalinity obtained from August 2020 cruise. The geographical distribution and variability of six buffer factors, which amended the existing Revelle factor, are discussed their relationship with the hydrological parameters of temperature and salinity. The calculated results of six buffer factors showed the spatial variations according to the distributions of various water masses. The buffer capacity was low in the East Sea Surface Mixed Water (ESMW) and South Sea Surface Mixed Water (SSMW) where upwelling occurred, and showed an intermediate value in the Yellow Sea Surface Water (YSSW). In addition, the buffer capacity increased in the order of high temperature Tsushima Warm Current (TWC) and Changjiang Diluted Water (CDW). This means that the Changjiang discharge water in summer strengthens the buffer capacity of the study area. The highest buffer capacity of CDW is due to its relatively higher temperature and biological productivity, and a summer stratification. Temperature showed a good positive correlation (R2=0.79) with buffer capacity in all water masses, whereas salinity exhibited a poor negative correlation (R2=0.30). High temperature strengthens buffer capacity through thermodynamic processes such as gas exchange and distribution of carbonate system species. In the case of salinity, the relationship with buffer capacity is reversed because salinity of the study area is not controlled by precipitation or evaporation but by a local freshwater input and mixing with upwelled water.

The responsibility of C-terminal domain in the thermolabile haemolysin activity of Vibrio parahaemolyticus and inhibition treatments by Phellinus sp. extracts

  • Tran Thi Huyen;Ha Phuong Trang;Nguyen Thi-Ngan;Bui Dinh-Thanh;Le Pham Tan Quoc;Trinh Ngoc Nam
    • Fisheries and Aquatic Sciences
    • /
    • v.26 no.3
    • /
    • pp.204-215
    • /
    • 2023
  • The thermolabile haemolysin (tlh) of Vibrio parahaemolyticus (Vptlh) from V. parahaemolyticus is a multiple-function enzyme, initially describes as a haemolytic factor activated by lecithin and phospholipase A2 enzymatic activity (Shinoda, 1991; Vazquez-Morado, 2021; Yanagase et al., 1970). Until now, the tlh structure has hypothesized including N-terminal and C-terminal domain, but what domain of the Vptlh structure does the haemolytic activity has not been refined yet. In this study, a 450-bp VpTLH nucleotide sequence of the entire Vptlh gene encoded the C-terminal domain cloned firstly to examine its responsibility in the activity of the Vptlh. The C-terminal domain fused with a 6-His-tag named the His-tag-VpC-terminal domain was expressed successfully in soluble form in the BL21 (DE3) PlysS cell. Remarkably, both expression and purification results confirmed a high agreement in the molecular weight of the His-tag-VpC-terminal domain was 47 kDa. This work showed the His-tag-VpC-terminal domain lysed the erythrocyte membranes in the blood agar and the phosphate buffered saline (0.9%) media without adding the lecithin substrate of the phospholipase enzyme. Haemolysis occurred at all tested diluted concentrations of His-tag-VpC-terminal domain (p < 0.05), providing evidence for the independent haemolytic activity of the His-tag-VpC-terminal domain. The content of 100 ㎍ of the His-tag-VpC-terminal domain brought the highest haemolytic activity of 80% compared to that in the three remaining contents. Significantly, the His-tag-VpC-terminal domain demonstrated not to involve the phospholipase activity in Luria-Bertani agar supplemented with 1% (vol/vol) egg yolk emulsion. All results proved the vital responsibility of the His-tag-VpC-terminal domain in causing the haemolytic activity without the required activation by the phospholipase enzyme. Raw extracts of Phellinus igniarus and Phellinus pipi at 10-1 mg/mL inhibited the haemolytic activity of the His-tag-VpC-terminal domain from 67.7% to 87.42%, respectively. Hence applying the His-tag-VpC-terminal domain as a simple biological material to evaluate quickly potential derivatives against the Vptlh in vivo conditions will accessible and more advantageous than using the whole of the Vptlh.

Phytoplankton Diversity and Community Structure Driven by the Dynamics of the Changjiang Diluted Water Plume Extension around the Ieodo Ocean Research Station in the Summer of 2020 (2020년 하계 장강 저염수가 이어도 해양과학기지 주변 해역의 식물플랑크톤 다양성 및 개체수 변화에 미치는 영향)

  • Kim, Jihoon;Choi, Dong Han;Lee, Ha Eun;Jeong, Jin-Yong;Jeong, Jongmin;Noh, Jae Hoon
    • Journal of the Korean Society of Marine Environment & Safety
    • /
    • v.27 no.7
    • /
    • pp.924-942
    • /
    • 2021
  • The expansion of the Changjiang Diluted Water (CDW) plume during summer is known to be a major factor influencing phytoplankton diversity, community structure, and the regional marine environment of the northern East China Sea (ECS). The discharge of the CDW plume was very high in the summer of 2020, and cruise surveys and stationary monitoring were conducted to understand the dynamics of changes in environmental characteristics and the impact on phytoplankton diversity and community structure. A cruise survey was conducted from August 16 to 17, 2020, using R/V Eardo, and a stay survey at the Ieodo Ocean Research Station (IORS) from August 15 to 21, 2020, to analyze phytoplankton diversity and community structure. The southwestern part of the survey area exhibited low salinity and high chlorophyll a fluorescence under the influence of the CDW plume, whereas the southeastern part of the survey area presented high salinity and low chlorophyll a fluorescence under the influence of the Tsushima Warm Current (TWC). The total chlorophyll a concentrations of surface water samples from 12 sampling stations indicated that nano-phytoplankton (20-3 ㎛) and micro-phytoplankton (> 20 ㎛) were the dominant groups during the survey period. Only stations strongly influenced by the TWC presented approximately 50% of the biomass contributed by pico-phytoplankton (< 3 ㎛). The size distribution of phytoplankton in the surface water samples is related to nutrient supplies, and areas where high nutrient (nitrate) supplies were provided by the CDW plume displayed higher biomass contribution by micro-phytoplankton groups. A total of 45 genera of nano- and micro-phytoplankton groups were classified using morphological analysis. Among them, the dominant taxa were the diatoms Guinardia flaccida and Nitzschia spp. and the dinoflagellates Gonyaulax monacantha, Noctiluca scintillans, Gymnodinium spirale, Heterocapsa spp., Prorocentrum micans, and Tripos furca. The sampling stations affected by the TWC and low in nitrate concentrations presented high concentrations of photosynthetic pico-eukaryotes (PPE) and photosynthetic pico-prokaryotes (PPP). Most sampling stations had phosphate-limited conditions. Higher Synechococcus concentrations were enumerated for the sampling stations influenced by low-nutrient water of the TWC using flow cytometry. The NGS analysis revealed 29 clades of Synechococcus among PPP, and 11 clades displayed a dominance rate of 1% or more at least once in one sample. Clade II was the dominant group in the surface water, whereas various clades (Clades I, IV, etc.) were found to be the next dominant groups in the SCM layers. The Prochlorococcus group, belonging to the PPP, observed in the warm water region, presented a high-light-adapted ecotype and did not appear in the northern part of the survey region. PPE analysis resulted in 163 operational taxonomic units (OTUs), indicating very high diversity. Among them, 11 major taxa showed dominant OTUs with more than 5% in at least one sample, while Amphidinium testudo was the dominant taxon in the surface water in the low-salinity region affected by the CDW plume, and the chlorophyta was dominant in the SCM layer. In the warm water region affected by the TWC, various groups of haptophytes were dominant. Observations from the IORS also presented similar results to the cruise survey results for biomass, size distribution, and diversity of phytoplankton. The results revealed the various dynamic responses of phytoplankton influenced by the CDW plume. By comparing the results from the IORS and research cruise studies, the study confirmed that the IORS is an important observational station to monitor the dynamic impact of the CDW plume. In future research, it is necessary to establish an effective use of IORS in preparation for changes in the ECS summer environment and ecosystem due to climate change.

Determination of Sulfur Requirement to Adjust pH of Alkaline Soil by Buffer Curve Method (알칼리성 토양 pH 교정시 완충곡선법을 이용한 황 시용량 결정)

  • Lee, In-Bog;Lim, Jae-Hyun;Yiem, Myoung-Soon
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.33 no.6
    • /
    • pp.405-415
    • /
    • 2000
  • To determine application rate of elemental sulfur to adjust pH of alkaline soil, buffer curve method was investigated. The elemental sulfur required to control pH 8.3 to pH 6.3 by buffer curve calculation was treated in two soils of silty loam and sandy loam, and the sulfur-mixed soils were moistened with 50% of water holding capacity during incubation of 6 weeks at $30^{\circ}C$. Soil pH was lowered with incubation and reached to target point after 4 weeks of incubation, and elemental sulfur was oxidised entirely to sulfate. This means that buffer curve has the accuracy to determine sulfur application rate in alkaline soil. However it is estimated that application rate of sulfur should be carefully determined in the field scale. Excess application of elemental sulfur resulted in extremely low soil pH and caused the hinderance of lettuce growth by nutritional imbalance and ion toxicity. To simplify the determination procedure of sulfur requirement, buffer curve method by addition of 0.1N-HCl solution as unit of mL was developed, it was compared with theroutine methods which diluted $H_2SO_4$ solution and $Ca(OH)_2$ are added as cmolc per kg soil to adjust each pH step. Buffer capacities, cmolc kg $soil^{-1}$ $pH^{-1}$, calculated from two buffer curves were not significantly different. The result indicates that buffer curve method by 0.1N-HCl can be used to adjust high pH of alkaline soil.

  • PDF

Determination of methamphetamine, 4-hydroxymethamphetamine, amphetamine and 4-hydroxyamphetamine in urine using dilute-and-shoot liquid chromatography-tandem mass spectrometry (시료 희석 주입 LC-MS/MS를 이용한 소변 중 메스암페타민, 4-하이드록시메스암페타민, 암페타민 및 4-하이드록시암페타민 동시 분석)

  • Heo, Bo-Reum;Kwon, NamHee;Kim, Jin Young
    • Analytical Science and Technology
    • /
    • v.31 no.4
    • /
    • pp.161-170
    • /
    • 2018
  • The epidemic of disorders associated with synthetic stimulants, such as methamphetamine (MA) and amphetamine (AP), is a health, social, legal, and financial problem. Owing to the high potential of their abuse and addiction, reliable analytical methods are required to detect and identify MA, AP, and their metabolites in biological samples. Thus, a dilute-and-shoot liquid chromatography-tandem mass spectrophotometry (LC-MS/MS) was developed for simultaneous determination of MA, 4-hydroxymethamphetamine (4HMA), AP, and 4-hydroxyamphetamine (4HA) in urine. Urine sample ($100{\mu}L$) was mixed with $50{\mu}L$ of mobile phase consisting of 0.4 % formic acid and methanol and $50{\mu}L$ of working internal-standard solution. Aliquots of $8{\mu}L$ diluted urine was injected into the LC-MS/MS system. For all analytes, chromatographic separation was performed using a C18 reversed-phase column with gradient elution and a total run time of 5 min. The identification and quantification were performed by multiple reaction monitoring (MRM). Linear least-squares regression was conducted to generate a calibration curve, with $1/x^2$ as the weighting factor. The linear ranges were 2.0-200, 1.0-800, and 10-2500 ng/mL for 4HA and 4HMA, AP, and MA, respectively. The inter- and intraday precisions were within 6.6 %, whereas the inter- and intraday accuracies ranged from -14.9 to 11.3 %. The low limits of quantification were 2.0 ng/mL (4HA and 4HMA), 1.0 ng/mL (AP), and 10 ng/mL (MA). The proposed method exhibited satisfactory selectivity, dilution integrity, matrix effect, and stability, which are required for validation. Moreover, the purification efficiency of high-speed centrifugation was clearly higher than 6-15 % for QC samples (n=5), which was higher than that of the membrane-filtration method. The applicability of the proposed method was tested by forensic analysis of urine samples from drug abusers.

Screening Methods for Plant-Coating Materials and Transpiration Inhibitory Effect of Soybean Oil to Crops (식물 코팅 소재 선발법과 작물들에 대한 콩 오일의 증산 억제 효과)

  • Jung, In Hong;Park, No Bong;Kim, Sang-Yeol;Na, Young-Eun;Kim, Soon-Il
    • Korean Journal of Plant Resources
    • /
    • v.27 no.4
    • /
    • pp.380-391
    • /
    • 2014
  • Plants as well as crops are damaged by a combination of the hot and dry winds that has been a major factor in the reduction of crop production. A means to protect them from damaging conditions is to consider a coating material. In this study, we established laboratory screening methods to find a coating material to protect a crop from rapid transpiration caused by various factors. In a test measuring the weight loss of kidney bean seedlings for 6 days, Avion treatments decreased its weight loss (P=0.05). Owing to long-time spend in completing this assay, we performed a more simple method using a cobalt chloride paper strip, which changes from blue to red colors under water condition. Beewax, guagum, paraffin liquid, soybean oil, and PE-635 gave a waterproofing effect above 37 and 43% at 0.5 and 1 h after treatment, respectively. However, these tested materials did not show significant waterproofing results at 2 h. Although the methods produced reasonable results, a screening method to obtain more objective data is needed. An alternative is to use an instrument that can detect the transpiration of crop leaves. In a preliminary test using barley leaves, a portable photosynthesis system showed transpiration inhibition of 2% soybean oil and 10 times-diluted Avion under field conditions. In another test using the leaves of maize seedlings and apricot tree, 2% liquid paraffin and plant oils such as apricot oil, linseed oil, olive oil, and soybean oil showed significant transpiration inhibition (P=0.05). Especially, paraffin liquid and soybean oil selected from above tests gave good transpiration inhibitory effects against rice at 2%. In addition, the mixture of 2% soybean oil and a spreader showed more elevated inhibition results comparing with soybean oil or the spreader alone indicating that the spreader may be attributed to more uniform diffusion of the hydrophobic material onto the leaf surface of maize seedlings. The hydrophobic material coated physically the stomata and cuticle layers on leaf surfaces of rice. These hydrophobic materials screened in this study are expected to be used as plant coating materials.

Measuring Intracellular Mycobacterial Killing Using a Human Whole Blood Assay (인체 전혈 모델을 이용한 세포내 결핵균 살균력에 관한 연구)

  • Cheon, Seon-Hee;Song, Ho-Yeon;Lee, Eun-Hee;Oh, Hee-Jung;Kang, In-Sook;Cho, Ji-Yoon;Hong, Young-Sun
    • Tuberculosis and Respiratory Diseases
    • /
    • v.53 no.5
    • /
    • pp.497-509
    • /
    • 2002
  • Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.