• Title/Summary/Keyword: Cocktail approach

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Comparison of Gastrointestinal Permeability of Caffeine, Propranolol, Atenolol, Ofloxacin, and Quinidine Measured Using Ussing Chamber System and Caco-2 Cell Monolayer

  • Song, Im-Sook;Choi, Young A;Choi, Min-Koo
    • Mass Spectrometry Letters
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    • v.8 no.2
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    • pp.34-38
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    • 2017
  • The purpose of this study was to develop a cocktail approach for the measurement of the permeability of marker compounds, caffeine and propranolol (high permeability), ofloxacin (intermediate), atenolol (low), and quinidine (P-glycoprotein substrate), simultaneously. Then we compared the permeability in Caco-2 cells with that in rat intestinal segments. The difference between individual measurement and cocktail approach was less than 20 %, and the permeabilities of these compounds were similar to those previously reported, suggesting that the cocktail transport study and simultaneous drug analysis were successfully developed and validated in this study. Additionally, in the application of this cocktail method, the permeability of five drugs in rat jejunum was similar to that in ileum but different from that in colon, which was measured using the Ussing chamber system. Moreover, permeability in jejunum and ileum was similar to that in Caco-2 cells. In conclusion, the permeability in Caco-2 cells was equivalent to the permeability in rat jejunum and ileum determined with the Ussing system. Therefore, this newly developed cocktail assay and its application to the Ussing system can be a useful tool for robust and rapid screening for site-specific permeability in rat intestine, thus accelerating the prediction of absorption of new chemical entities.

On the intra-granular behaviour of a cocktail of inert gases in oxide nuclear fuel: Methodological recommendation for accelerated experimental investigation

  • Romano, M.;Pizzocri, D.;Luzzi, L.
    • Nuclear Engineering and Technology
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    • v.54 no.5
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    • pp.1929-1934
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    • 2022
  • Besides recent progresses in the physics-based modelling of fission gas and helium behaviour, the scarcity of experimental data concerning their combined behaviour (i.e., cocktail) hinders further model developments. For this reason, in this work, we propose a modelling methodology aimed at providing recommendations for accelerated experimental investigations. By exploring a wide range of annealing temperatures and cocktail compositions with a physics-based modelling approach we identify the most interesting conditions to be targeted by future experiments. To corroborate the recommendations arising from the proposed methodology, we include a sensitivity analysis quantifying the impact of the model parameters on fission gas and helium release, in conditions representative of high and low burnup.

Supplementation of enzyme cocktail in chickens diet is an effective approach to increase the utilization of nutrient in wheat-based diets

  • Ko, Hanseo;Kang, Hwan Ku;Moturi, Joseph;Ingale, Santosh Laxman;Kim, Jinsoo
    • Journal of Animal Science and Technology
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    • v.63 no.1
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    • pp.69-76
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    • 2021
  • This experiment was conducted to evaluate the effect of supplementing enzyme cocktail on growth performance, digestibility of nutrients, and monosaccharide concentration in ileum and ceca of broiler chickens fed wheat-based diets. A total of 600 male broilers (42.26 ± 1.76 g, 0 day old) were used for 35 days of feeding trial consisting of 2 phases (starter phase from d 0 to 21 and finisher phase from d 21 to 35). Four dietary treatments were prepared based on wheat diets containing four levels of enzyme cocktail supplementation at 0, 0.2, 0.3, and 20 g/kg. Overall, dietary enzyme cocktail supplementation decreased feed conversion ratio (linear p = 0.007; quadratic p = 0.013) and improved (linear p < 0.05) the apparent ileal digestibility of dry matter (DM), crude protein, and soluble and insoluble non-starch polysaccharides. The apparent total tract digestibility of DM and gross energy were increased (linear p < 0.01) with increasing supplementation levels of the dietary enzyme cocktail. The concentrations of arabinose, xylose, mannose, and glucose in ileal digesta were linearly increased (p < 0.01) with increasing enzyme cocktail supplementation levels. In addition, the quadratic effect was observed (quadratic p = 0.046) in mannose concentration of ileal digesta. The concentration of arabinose, xylose, mannose, and galactose in cecal digesta was increased (linear p < 0.05) with increasing dietary enzyme cocktail supplementation levels. The supplementation of enzyme cocktail efficiently increased the utilization of nutrients in broiler and there was no adverse effects of high dosage supplementation level.

Effects of dietary enzyme cocktail on diarrhea and immune responses of weaned pigs

  • Kang, Joowon;Cho, Jeeyeon;Jang, Kibeom;Kim, Junsu;Kim, Sheena;Mun, Daye;Kim, Byeonghyeon;Kim, Younghwa;Park, Juncheol;Choe, Jeehwan;Song, Minho
    • Korean Journal of Agricultural Science
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    • v.44 no.4
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    • pp.525-530
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    • 2017
  • Weaning is the most stressful event for nursery pigs because they are moved from familiar to unfamiliar environments. In addition, weaned pigs have immature digestive and immune systems. This situation makes weaned pigs susceptible to diseases and makes the absorption of nutrients from diets difficult. A feed approach, such as dietary enzyme supplementation, can be considered a solution. This study investigated the effects of dietary enzyme cocktail on diarrhea and immune responses of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.05% enzyme cocktail (Cocktail; combination of xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Incidence of diarrhea, packed cell volume (PCV), white blood cells (WBC) count, and immunoglobulin content were measured. A significantly lower incidence of diarrhea (p < 0.05) was observed in the Cocktail group as compared with the CON group. The Cocktail group also showed a decreased PCV (p < 0.1) on d 3 after weaning than the CON group. However, no differences were observed for number of WBC and contents of immunoglobulin G, M, and A between the Cocktail and CON groups. Consequently, inclusion of an enzyme cocktail in diets for weaned pigs had a positive influence on gut health by reducing the incidence of diarrhea in the present study.

A Study on Fire Performance and Heat Transfer of HPC Column with Fiber-Cocktail in ISO Fire under Loading Condition (표준화재 재하조건 Fiber Cocktail을 혼입한 고강도 콘크리트 기둥의 전열 특성 및 화재 거동에 관한 연구)

  • Kim, Heung-Youl;Kim, Hyung-Jun;Jeon, Hyun-Kyu;Youm, Kwang-Soo
    • Journal of the Korea Concrete Institute
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    • v.22 no.1
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    • pp.29-39
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    • 2010
  • In this study, experimental test and numerical analysis were conducted to investigate the heat transfer characteristics and fiber performance of high strength concrete. The fire characteristics of the high strength concrete that couldn't be obtained through the test due to specific requirements and restrictions were forecast using numerical analysis approach. The outcome from the numerical analysis and the test were compared to verify and improve the reliability of the analysis. A numerical analysis of 80 and 100 MPa high strength concrete cases were carried out to identify the heat transfer characteristics and fire behavior using software, ABACUS (V6.8) From the results of verification experiment, a 25~55% level of beam shrinkage reduction was observed compared to the concrete without Fiber-Cocktail, indicating the improved fire resistance performance, which appeared to be attributable to the function of Fiber-Cocktail that was able to control the heat transfer characteristics and ultimately result in enhancing the fire resistance performance.

Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.88-89
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    • 2013
  • A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

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A Synthetic Tul4 and FopA Peptide Cocktail of Francisella tularensis Induces Humoral and Cell-Mediated Immune Responses in Mice

  • Oh, Hanseul;Kim, C-Yoon;Kim, Chang-Hwan;Hur, Gyeung-Haeng;Park, Jae-Hak
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1613-1619
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    • 2016
  • Francisella tularensis is a highly virulent pathogen of humans and other mammals. Moreover, F. tularensis has been designated a category A biothreat agent, and there is growing interest in the development of a protective vaccine. In the present study, we determine the in vitro and in vivo immune responses of a subunit vaccine composed of recombinant peptides Tul4 and FopA from epitopes of the F. tularensis outer membrane proteins. The recombinant peptides with adjuvant CpG induced robust immunophenotypic change of dendritic cell (DC) maturation and secretion of inflammatory cytokines (IL-6, IL-12). In addition, the matured DCs enabled ex vivo proliferation of naive splenocytes in a mixed lymphocyte reaction. Lastly, we determined the in vivo immune response by assessment of antibody production in C57BL/6 mice. Total IgG levels were produced after immunization and peaked in 6 weeks, and moreover, Tul4-specific IgG was confirmed in the mice receiving peptides with or without CpG. Based on these results, we concluded that the recombinant peptides Tul4 and FopA have immunogenicity and could be a safe subunit vaccine candidate approach against F. tularensis.

Transcriptome analysis and promoter sequence studies on early adipogenesis in 3T3-L1 cells

  • Kim, Su-Jong;Lee, Ki-Hwan;Lee, Yong-Sung;Mun, Eun-Gyeng;Kwon, Dae-Young;Cha, Youn-Soo
    • Nutrition Research and Practice
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    • v.1 no.1
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    • pp.19-28
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    • 2007
  • To identify regulatory molecules which play key roles in the development of obesity, we investigated the transcriptional profiles in 3T3-L1 cells at early stage of differentiation and analyzed the promoter sequences of differentially regulated genes. One hundred and sixty-one (161) genes were found to have significant changes in expression at the 2nd day following treatment with differentiation cocktail. Among them, 86 transcripts were up-regulated and 75 transcripts were down-regulated. The 161 transcripts were classified into 10 categories according to their functional roles; cytoskeleton, cell adhesion, immune, defense response, metabolism, protein modification, protein metabolism, regulation of transcription, signal transduction and transporter. To identify transcription factors likely involved in regulating these differentially expressed genes, we analyzed the promoter sequences of up- or - down regulated genes for the presence of transcription factor binding sites (TFBSs). Based on coincidence of regulatory sites, we have identified candidate transcription factors (TFs), which include those previously known to be involved in adipogenesis (CREB, OCT-1 and c-Myc). Among them, c-Myc was also identified by our microarray data. Our approach to take advantage of the resource of the human genome sequences and the results from our microarray experiments should be validated by further studies of promoter occupancy and TF perturbation.

Radioimmunoscintigraphy Using IMACIS-1 in Gastrointestinal Cancer (IMACIS-1을 이용한 위장관 종양의 방사면역신티그램)

  • Sohn, Hyung-Sun;Kim, Choon-Yul;Bahk, Yong-Whee
    • The Korean Journal of Nuclear Medicine
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    • v.24 no.1
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    • pp.29-36
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    • 1990
  • Most of the diagnostic methods currently used for the detection of neoplastic masses provide indirect evidence. To obtain greater specificity in the interpretation of neoplasias by in vivo methods, the immunological approach appears to be most promising. Two problems that interfered with progress in this field were the lack of tumor specific antigen and the lack of well-defined and reproducible antibodies. To improve the sensitivity and specificity of radioimmunoscintigraphy as a technique for tumor localization, the use of monoclonal antibodies, fragments of antibodies and single photon emission computerized tomography (SPECT) are reasonable. The obvious advantages of monoclonal antibodies are their homogeneity, their specificity for the immunizing antigen and the reaction with a single determinant-thus no large immunecomplexes with antigen are formed. Monoclonal antibody technique has recently provided an opportunity to reevaluate the role of nuclear medicine for the diagnosis of malignant diseases by using the immunological approach. Out first results by means of radioimmunoscintigraphy of CEA and CA 19-9 producing tumors using a cocktail of fragments F $(ab')_2$, of mocolonal antibodies to CA 19-9 and CEA labeled with $^{131}I$ (IMACIS-1) are reported. The aims of this investigation was to evaluate the role of immunoscintigraphy in patients with colorectal and other cancers for diagnosis of local recurrences and metastasis. This report contains results of the first 8 colorectal and pancreas cancer patients with the elevation of the level of serum CEA and/or CA 19-9. IMACIS-1 was injected intravenously during 30 minutes in 100 ml saline solution after skin test. Planar scintigrams were recorded 3, 5 and 7 days after the injection of the IMACIS-1. Anterior, lateral and posterior views of the liver as well as anterior and posterior views of the pelvis were obtained in each patients as an $^{131}I-antibody$ image. We were able to localize exactly the malignant process with the double-nuclide double-compound $^{99m}Tc\;^{131}I$ (Tc+l) scintigrams. In Tc & I double-nuclide scintigraphy, computer subtraction display provided more clear localization of the tumor. We compared the results of radioimmunoscintigraphy with CT, ultrasonograms, conventional scintigrams. The results were as follows: 1) The sensitivity and specificity of radioimmunoscintigraphy using the fragments $F(ab')_2$ of the cocktails of CEA and CA 19-9 monoclonal antibodies were 80% and 100% respectively. 2) Tumor detection rate was not proportionated to the level of serum tumor markets. 3) Second tracer technique was essential for tumor localization as an anatomic landmark using double-nuclide scintigraphy. 4) A slow infusion of the antibodies was necessary to prevent the formation of large immune complexes. 5) Tumor/non-tumor radioactivity was most elevated at 7 days delayed imaging. 6) Using planar scintigraphic technique of $^{131}I$ labeled monoclonal antibodies are possible for imaging most of the tumors.

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Accuracy and Clinical Outcomes of Ultrasound-guided Glenohumeral joint Injection: Acromioclavicular Approach in Patients with Adhesive Capsulitis (초음파를 이용한 견관절 관절강내 주사의 정확성과 임상적 결과: 유착성 관절낭염 환자의 견봉 쇄골 접근법)

  • Lhee, Sang-Hoon;Hwang, Seok-Min
    • The Journal of Korean Orthopaedic Ultrasound Society
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    • v.5 no.1
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    • pp.9-14
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    • 2012
  • Purpose: The purpose of the study was to evaluate the accuracy and clinical outcome of ultrasound-guided glenohumeral joint steroid injection on adhesive capsulitis. Materials and Methods: Patients who were diagnosed as adhesive capsulitis by MRI and physical examination and did not improve their symptom with physical therapy and NSAIDS treatment more than 6 months were included in the study. Patients who showed any other shoulder pathology or history if trauma were excluded from the study. 33 patients including 15 males and 18 females were enrolled in the study, the average age being 55.1 (age 42~72). Cocktail of steroid, lidocaine, saline and contrast medium injected inside shoulder glenohumeral joint using novel approach (which we called acromioclavicular approach) under ultrasound guidance. Clinical outcome was measured through passive range of motion and VAS scoring system. Results: Based on radiographic findings, cases were classified according to the leakage of contrast medium; perfect confinement of contrast-medium inside the capsule, partial leakage of the medium and contrast-medium found at outside the joint. Total 25 cases (76%) out of 33 cases showed perfect confinement of contrast-medium inside the glenohumeral joint. Partial leakage was observed in 6 cases (18%), and contrast medium was observed outside of the glenohumeral joint in 2 cases (6%). Perfect-confinement group showed $111^{\circ}$($80{\sim}140^{\circ}$) of forward flexion and $48^{\circ}$($0{\sim}90^{\circ}$) of external rotation before injection, and improved to $134^{\circ}$($90{\sim}150^{\circ}$) of forward flexion and $70^{\circ}$($30{\sim}90^{\circ}$) of external rotation after injection (p<0.01). Partial leakage showed $120^{\circ}$($90{\sim}150^{\circ}$) of forward flexion and $70^{\circ}$($10{\sim}90^{\circ}$) of external rotation before injection, and improved to $139^{\circ}$($135{\sim}140^{\circ}$) of forward flexion and $78^{\circ}$($50{\sim}90^{\circ}$) of external rotation after injection (p<0.01). VAS score improved from 7.1 (score 3~9) to 2.6 (score 0~5) (p<0.01) in perfect confinement group, from 7.5 (score 7~9) to 3.3 (score 2~4) (p<0.01) in partial leakage group. Two group showed no significant difference. Conclusion: Accuracy of Acromioclavicular approach was 94% which is better than any other methods published so far. Partial leakage of the injection material did not show inferior result compared to perfect injection.

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