• Title/Summary/Keyword: Immunoassays

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Analytical-numerical formula for estimating the characteristics of a cylindrical NaI(Tl) gamma-ray detector with a side-through hole

  • Thabet, Abouzeid A.;Badawi, Mohamed S.
    • Nuclear Engineering and Technology
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    • v.54 no.10
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    • pp.3795-3802
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    • 2022
  • NaI(Tl) scintillation materials are considered to be one of many materials that are used exclusively for γ-ray detection and spectroscopy. The gamma-ray spectrometer is not an easy-to-use device, and the accuracy of the numerical values must be carefully checked based on the rules of the calibration technique. Therefore, accurate information about the detection system and its effectiveness is of greater importance. The purpose of this study is to estimate, using an analytical-numerical formula (ANF), the purely geometric solid angle, geometric efficiency, and total efficiency of a cylindrical NaI(Tl) γ-ray detector with a side-through hole. This type of detector is ideal for scanning fuel rods and pipelines, as well as for performing radio-immunoassays. The study included the calculation of the complex solid angle, in combination with the use of various points like gamma sources, located axially and non-axially inside the through detector side hole, which can be applied in a hypothetical method for calibrating the facility. An extended γ-ray energy range, the detector, source dimensions, "source-to-detector" geometry inside the side-through hole, path lengths of γ-quanta photons crossing the facility, besides the photon average path length inside the detector medium itself, were studied and considered. This study is very important for an expanded future article where the radioactive point source can be replaced by a volume source located inside the side-trough hole of the detector, or by a radioactive pipeline passing through the well. The results provide a good and useful approach to a new generation of detectors that can be used for low-level radiation that needs to be measured efficiently.

Biodegradation of marine microplastics by the whole-cell catalyst overexpressing recombinant PETase (PET분해효소(PETase) 과발현 전세포 촉매의 해양미세플라스틱 생분해 활성 연구)

  • Hyunji, Kim;Jong-Ha, Park;Ae-Ran, Park;Dae-Hee, Lee;Joonho, Jeon;Hyuk Taek, Kwon;Sung In, Lim
    • Journal of Marine Bioscience and Biotechnology
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    • v.14 no.2
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    • pp.133-142
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    • 2022
  • The increased production and consumption of polyethylene terephthalate (PET)-based products over the past several decades has resulted in the discharge of countless tons of PET waste into the marine environment. PET microparticles resulting from the physical erosion of general PET wastes end up in the ocean and pose a threat to the marine biosphere and human health, necessitating the development of new technologies for recycling and upcycling. Notably, enzyme-mediated PET degradation is an appealing option due to its eco-friendly and energy-saving characteristics. PETase, a PET-hydrolyzing enzyme originating from Ideonella sakaiensis, is one of the most thoroughly researched biological catalysts. However, the industrial application of PETase-mediated PET recycling is limited due to the low stability and poor reusability of the enzyme. Here we developed the whole-cell catalyst (WCC) in which functional PETase is attached to the outer membrane of Escherichia coli. Immunoassays are used to identify the surface-expressed PETase, and we demonstrated that the WCC degraded PET microparticles most efficiently at 30℃ and pH 9 without agitation. Furthermore, the WCC increased the PET-degrading activity in a concentration-dependent manner, surpassing the limited activity of soluble PETase above 100 nM. Finally, we demonstrated that the WCC could be recycled up to three times.

Detection of a Thermal Stable-Soluble Protein (TSSP) as a Marker of Peanut Adulteration Using a Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay based on Monoclonal Antibodies

  • Sol-A Kim;Sazzad Hossen Toushik;Jeong-Eun Lee;Won-Bo Shim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.9
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    • pp.1170-1178
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    • 2023
  • Food allergy represents a severe problem for many societies, including sensitive populations, academies, health authorities, and the food industry. Peanut allergy occupies a special place in the food allergy spectrum. To prevent consumption by consumers suffering from a peanut allergy, a rapid and sensitive detection method is essential to identify unintended peanut adulteration in processed foods. In this study, we produced four monoclonal antibodies (MAbs; RO 3A1-12, PB 4C12-10, PB 5F9-23, and PB 6G4-30) specific to thermo-stable and soluble proteins (TSSPs) of peanut and developed an enzyme-linked immunosorbent assay (ELISA) based on the MAbs. Among them, PB 5F9-23 MAb was firmly bound to Ara h 1, and other MAbs strongly reacted to Ara h 3 in the Western blot analysis. An antibody cocktail solution of the MAbs was used to enhance the sensitivity of an indirect ELISA, and the limit of detection of the indirect ELISA based on the antibody cocktail solution was 1 ng/ml and improved compared to the indirect ELISA based on the single MAb (11 ng/ml). The cross-reaction analysis revealed the high specificity of developed MAbs to peanut TSSPs without cross-reaction to other food allergens, including nuts. Subsequently, analyzing processed foods by indirect ELISA, all foods labeled as containing peanuts in the product description were confirmed to be positive. The results indicate that the developed antibodies exhibit high specificity and sensitivity to peanuts and can be used as bio-receptors in immunoassays or biosensors to detect intentional or unintentional adulteration of peanuts in processed foods, particularly heat-processed foods.

Inhibitory Effect of Jeungaektang Water Extract on Nitric Oxide and Cytokine Production in Lipopolysaccharide - activated RAW 264.7 Cells (증액탕(增液湯) 물추출물이 LPS로 유도된 RAW 264.7 cell에서의 Nitric Oxide 및 Cytokine에 미치는 영향)

  • Ahn, Sun-June;Lee, Jong-Rok;Kim, Sang-Chan;Jee, Seon-Young
    • Herbal Formula Science
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    • v.15 no.1
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    • pp.163-173
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    • 2007
  • Jeungaektang (JAT) is the herbal formula, has the effect of moistening the dryness by activating lung Qi and by nourishing Yin, has being used for dryness syndromes. Generally the herbal formulae for moistening dryness are used for exogenous or endogenous dryness syndromes. JAT has been clinically used for the treatment of endogenous dryness syndromes. It is composed of Scrophulariae Radix. Rehmanniae Radix and Liriopis Tuber. Recent studies showed that JAT has a protective effect against $CCl_{4}-induced$ hepatotoxicity and anti-inflammatory effects against ear swelling of mouse induced by Crotonis Fructus. However, the effect of JAT on the immunological activity was rarely studied. Therefore, this study evaluated the effects of JAT the regulatory mechanism of nitric oxide (NO) and cytokines in the lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. After the treatment of JAT water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Cyclooxygenase-2 (COX -2) and inducible nitric oxide synthase (iNOS) were determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidence that JAT inhibited the production of nitrite and nitrate ($0.1{\sim}1.0$ mg/ml), iNOS ($0.1{\sim}1.0$ mg/ml), $interleukin-1{\beta}$ ( $0.1{\sim}1.0$ mg/ml) and tumor necrosis $factor-{\alpha}$ ($0.1{\sim}1.0$ mg/ml) in RAW 264.7 cells activated with LPS. Furthermore, JAT inhibited the expression of COX-2 expression and production of prostagladin E2 ($0.1{\sim}1.0$ mg/ml). These findings suggest that JAT can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

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Inhibitory Effect of Phellinus Igniarius water extract on TNF-$\alpha$, IL-1$\beta$, IL-6 and Nitric Oxide Production in lipopolysaccharide - activated Raw 264.7 cells (상황 물추출물이 LPS로 유도된 Raw 264.7 cell에서의 TNF-$\alpha$, IL-1$\beta$, IL-6 및 Nitric Oxide production에 미치는 영향)

  • Kim Sang Chan;Jung Youn Suk;Lee Jae Ryung;Kim Young Woo;Byun Boo Hyeong;Kwon Teag Kyu;Suh Seong Il;Byun Sung Hui;Kwon Young Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.880-886
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    • 2004
  • Phellinus igniarius has been clinically used for the treatment of hemorrhoidal fistula, dysmenorrhea and the prevention of cancer in traditional oriental medicine. Recent studies showed that Phellinus igniarius produced anti-cancer, anti-metastasis and immuno-modulatory effects, There is lack of studies regarding the effects of Phellinus igniarius on the immunological activities. The present study was conducted to evaluate the effect of Phellinus igniarius on the regulatory mechanism of cytokines and nitric oxide (NO) for the immunological activities in Raw 264,7 cells. After the treatment of Phellinus igniarius water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. COX-2 and iNOS were determined by Immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidence that Phellinus igniarius inhibited the production of nitrite and nitrate (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β) and interleukin-6 (IL-6), and the activation of phospholylation of inhibitor κBα (p-IκBα) in Raw 264.7 cells activated with lipopolysaccharide (LPS). These findings suggest that Phellinus igniarius can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

Expression and Significance of ER, PR, VEGF, CA15-3, CA125 and CEA in Judging the Prognosis of Breast Cancer

  • Zhang, Su-Jie;Hu, Yi;Qian, Hai-Li;Jiao, Shun-Chang;Liu, Zhe-Feng;Tao, Hai-Tao;Han, Lu
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3937-3940
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    • 2013
  • Objective: To explore the expression and significance of estrogen receptor (ER), progestrone receptor (PR), vascular endothelial growth factor (VEGF), CA15-3, CA125 and carcinoma embryonic antigen (CEA) expression in judging the prognosis of breast cancer. Materials and Methods: Sixty-five patients with breast cancer undergoing operations in the general surgery department were considered as the observation group, and 50 healthy outpatients of our hospital as the control group. Cubital venous blood was drawn in the morning from fasting patients in the two groups and chemiluminescence immunoassays were used to detect the levels of CA15-3, CA125 and CEA in serum. The follow-up duration was from 4 months to 2 years, and change in levels of the indicators was detected by dynamically drawing blood. After surgery, cancer tissue samples of patients in observation group remained on file (the non-recurrent patients were biopsied). Immunohistochemistry was applied to determine the expression of ER, PR and VEGF in tissue. Results: The effective rate of 12 patients with negative ER and PR expression was 33.3% in the observation group, being associated with prognosis to varying extents. Serum CA15-3, CA125 and CEA in the observation group were all significantly higher than in control group (p<0.01). With increase in pathological staging, levels of serum CA15-3, CA125 and CEA gradually increased (p<0.01). Levels in patients with lymph node metastasis were markedly higher than in those without (p<0.01). In addition, values with distal lymph node metastasis were notably higher than with adjacent lymph node metastasis (p<0.01). The postoperative follow-up results revealed that positive VEGF and levels of serum VEGF, CA15-3, CA125 and CEA in recurrence group were obviously higher than in non-recurrence group (p<0.01). Conclusions: Joint detection of ER and PR expression as well as levels of serum VEGF, CA15-3, CA125 and CEA is meaningful and can guide the diagnosis and treatment for breast cancer.

Evaluation of the antibodies against Pseudomonas tolaasii and Pseudomonas agarici for Immunoassays and Its Application (항 Pseudomonas tolaasii 및 Pseudomonas agarici 항체의 면역측정법을 위한 평가와 그 이용)

  • Jeon, Nak-Beom;Lee, Hyang-Burm;Shon, Dong-Hwa;Yu, Seung Hun
    • Research in Plant Disease
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    • v.9 no.2
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    • pp.94-98
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    • 2003
  • To produce antibodies against Pseudomonas tolaasii and P. agarici, lyophilized P. tolaasii and P. agarici ($5{\times}10^7$ cfu/ml) and Freund, s adjuvant were immunized into rabbits 4 times. The specificity and sensitivity of the antibodies were evaluated by immunodiffusion test and indirect enzyme-linked immunosorbent assay (id ELISA). The ${\alpha}$-P. tolaasii antibody was very specific only against P. tolaasii, while ${\alpha}$-P. agarici antibody was not specific and showed a high cross reactivity toward P. tolaasii with detection limit concentration of $2{\times}10^3$ cfu/ml. However, the cross reactivities of ${\alpha}$-P. agarici antibody toward the related species including P. reactans were very low. Our results showed that ${\alpha}$-P. tolaasii and ${\alpha}$-P. agarici antibodies against P. tolaasii and P. agarici, respectively, might be useful for rapid and simple detection of the causal agents of bacterial brown and yellow blotches in cultivated oyster mushrooms.

Evaluation of Pregnancy and Thyroid Function (임신과 갑상선 기능의 평가)

  • Park, Chang-Eun
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.1
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    • pp.1-10
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    • 2018
  • During early pregnancy, before the development of a functioning thyroid gland, thyroid stimulating hormone (TSH) is a very sensitive marker of thyroid dysfunction during pregnancy. Normal values have been modified during gestation with a downward shift. The fetus is influenced by the TSH supplied by the mother. TSH and free thyroxine (FT4) concentrations vary during pregnancy and conventional units can vary between laboratories. A downward shift of the TSH reference range occurs during pregnancy, with a decrease in both the lower and upper limits of maternal TSH, relative to the typical non-pregnant TSH reference range. Each laboratory produces its own reference TSH and FT4 concentrations because there are many different assays that yield different results in pregnancy. Therefore, automated immunoassays used for serum FT4 analysis are still used widely, but the important considerations discussed above must be noted. The use of population-based, trimester-specific reference ranges remains the best way to handle this issue The slight downward shift in the upper reference range of TSH occurring in the latter first trimester (7~12 weeks) of pregnancy, typically not observed prior to 7 weeks. Their use indicates high or low levels in a quantitative manner independent of the reference ranges. These data highlight the importance of calculating population-based pregnancy-specific thyroid parameter reference intervals. A precision medicine initiative in this area will require the collection and analysis of a large number of genetic, biological, psychosocial, and environmental variables in large cohorts of individuals. Large prospective randomized controlled trials will be needed to resolve these controversies.

Thermal Stable Soluble Proteins in Pork Fat and Meat, and Their Antigenicity (돈지육 및 돈육 중 열안정성 단백질의 존재 확인과 이의 항원성 확인)

  • Kim, Jeong-Sook;Lee, Jeong-Eun;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.31 no.6
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    • pp.445-450
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    • 2016
  • Thermal-stable soluble proteins (TSSP) in livestock products has been recently reported. Therefore, the development of antibodies and immunoassay using a TSSP is useful because the presence of TSSP can be measured on processed food. In this study, the existence of TSSPs in pork fat and meat was confirmed and their antigenicity was investigated. The extracts from pork fat and meat by heating method were analyzed by SDS-PAGE with 5% stacking and 12% separating gels. The protein profiles from the raw pork fat and meat extracts (major band ranged 25 to 100 kDa) without cooking and heating treatments were significantly different compared to those from cooked and heated pork fat and meat extracts (several major bands > 100 kDa and < 30 kDa). This meant that non thermal-stable soluble proteins ranged from 25 to 100 kDa may be denaturated to insoluble proteins by cooking and heating treatments, and TSSPs were in pork fat and meat at kept their properties. The confirmed TSSPs were used as an immunogen to investigate their antigenicity. Eight mice (5 mice for pork fat and 3 mice for pork meat) were separately immunized with the TSSPs of pork fat and meat, and the anti-sera obtained from the immunized mice showed high titer values. Polyclonal antibodies against each target protein showed the specific reaction to pork fat and meat, individually. These indicated that TSSP could be used as an immunogen to produce antibodies such as monoclonal and polyclonal antibodies. In addition, antibodies specific to TSSP from pork fat and meat may be used as a bio-receptor in immunoassays for the identification of fraudulent adulteration with pork fat and meat in livestock products.

Effects of Salviae Miltiorrhizae Radix Hot Aqueous Extract on Nitric Oxide and Prostaglandin E2 Production and on 1,1-diphenyl-2-picryl hydrazyl Radical Scavenging in Macrophages

  • Yeo, In Ho;Lee, Cham Kyul;Lee, Eun Yong
    • Journal of Pharmacopuncture
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    • v.17 no.1
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    • pp.7-12
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    • 2014
  • Objectives: The objective of this study is to investigate the effects of Salviae Miltiorrhizae Radix hot aqueous extract on nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production and on 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical scavenging in macrophages. Methods: Salviae Miltiorrhizae Radix (300 g) was heated at $100^{\circ}C$ with distilled water (2 L) for 4 hours. The extract was filtered and concentrated to 100 mL by using a rotary evaporator, was frozen at $-80^{\circ}C$, and was then freeze-dried by using a freezing-drying system. The RAW 264.7 macrophage was subcultured by using $10-{\mu}g/mL$ lipopolysaccharide (LPS). In order to evaluate cytotoxicity, we performed 3-(4,5-dimrthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and measured the cell viability. The NO production was measured by using Griess assays, and the $PGE_2$ production was measured by using enzyme immunoassays. The antioxidant activity, the 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical scavenging capability, was measured by using the DPPH method. Results: Cell viability with the 1-, 5-, 25-, 125- and $625-{\mu}g/mL$ Salviae Miltiorrhizae Radix hot aqueous extract was not significantly decreased compared to the cell viability without the extract. When 125 and $625{\mu}g/mL$ of Salviae Miltiorrhizae Radix hot aqueous extract were used, nitric oxide (NO) production in LPS-stimulated RAW 264.7 macrophages was significantly inhibited compared to that in the control group. When 25, 125, and $625{\mu}g/mL$ of Salviae Miltiorrhizae Radix hot aqueous extract were used, $PGE_2$ production in LPS-stimulated RAW 264.7 macrophages was significantly inhibited compared to that in the control group. The 125- and $625-{\mu}g/mL$ Salviae Miltiorrhizae Radix hot aqueous extracts had high DPPH free-radical scavenging capabilities in RAW 264.7 macrophages. Conclusion: This study indicates that Salviae Miltiorrhizae Radix hot aqueous extract suppresses NO and $PGE_2$ production and improves DPPH free-radical scavenging capability. Thus, it seems that Salviae Miltiorrhizae Radix hot aqueous extract may have an anti-inflammation effect and antioxidant activity.