• Title/Summary/Keyword: enzyme-linked immunosorbent assay(ELISA)

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Significance of Suppressor of Cytokine Signaling-3 Expression in Bladder Urothelial Carcinoma in Relation to Proinflammatory Cytokines and Tumor Histopathological Grading

  • Gaballah, Hanaa Hibishy;Shafik, Noha Mohamed;Wasfy, Rania Elsayed;Farha, Mohamed Osama Abou
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.1
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    • pp.307-314
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    • 2015
  • Background: Bladder cancer is among the five most common malignancies worldwide. Altered expression of suppressor of cytokine signaling -3 (SOCS-3) has been implicated in various types of human cancers; however, its role in bladder cancer is not well established. Aim: The present study was undertaken to investigate the mRNA expression of SOCS-3 in normal and cancerous bladder tissue and to explore its correlation with urinary levels of some proinflammatory cytokines, cytokeratin-18 (CK -18) and with tumor histopathological grading, in order to evaluate their role as potential diagnostic markers. Materials and Methods: SOCS3 mRNA expression levels were evaluated using quantitative real time PCR. Urinary levels of interleukins 6 and 8 were estimated by enzyme linked immunosorbent assay (ELISA). Cytokeratin-18 expression was analyzed by immuunohistochemistry then validated by ELISA. Results: SOC3 m RNA expression levels were significantly lower in high grade urothelial carcinoma ($0.36{\pm}0.12$) compared to low grade carcinoma ($1.22{\pm}0.38$) and controls ($4.08{\pm}0.88$), (p<0.001). However, in high grade urothelial carcinoma the urinary levels of IL-6, IL-8, total CK-18($221.33{\pm}22.84pg/ml$, $325.2{\pm}53.6pg/ml$, $466.7{\pm}57.40U/L$ respectively) were significantly higher than their levels in low grade carcinoma ($58.6{\pm}18.6pg/ml$, $58.3{\pm}50.2pg/ml$, $185.5{\pm}60.3U/L$ respectively) and controls ($50.9{\pm}23.0pg/ml$, $7.12{\pm}2.74pg/ml$, $106.7{\pm}47.3U/L$ respectively), (p<0.001). Conclusions: Advanced grade of urothelial bladder carcinoma is significantly associated with lowered mRNA expression of SOC3 as well as elevated urinary levels of proinflammatory cytokines and CK-18. Furthermore, our results suggested that urinary IL-8, IL-6 and CK-18 may benefit as noninvasive biomarkers for early detection as well as histopathological subtyping of urothelial carcinoma.

CA 15-3 (Mucin-1) and Physiological Characteristics of Breast Cancer from Lahore, Pakistan

  • Begum, Mumtaz;Karim, Sajjad;Malik, Arif;Khurshid, Rukhshan;Asif, Muhammad;Salim, Asmat;Nagra, Saeed Ahmed;Zaheer, Ahmad;Iqbal, Zafar;Abuzenadah, Adel Mohammed;Alqahtani, Mohammed Hussain;Rasool, Mahmood
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5257-5261
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    • 2012
  • Background: High incidence of breast cancer and its fatal effect has reached an alarming stage across the globe, including the third world countries. Many factors have been reported to be associated with the development of breast cancer but detailed structural and functional information is missing. CA 15-3 is one of the known potential tumor marker of breast cancer; however little is known about structure and functional site of this protein. Present study aims to investigate the functional role of CA 15-3 in breast cancer, especially in development and metastasis. Material and Methods: Hundred female breast cancer patients confirmed by histopathological reports were included in the study. Their physiological characters were recorded in a performa. Enzyme linked immunosorbent assay (ELISA) technique was used to estimate serum CA 15-3 level. Immunohistochemistry was done for estrogen (ER), progesterone (PR) and Her2/neu receptors expression. Results: The study revealed the details of physiological characteristics of female breast cancer. Mean age was $37.72{\pm}5.99$ and $55.05{\pm}7.28$ years and serum CA 15-3 (MUC1) level was $60.47{\pm}8.59$ and $63.17{\pm}4.58$ U/ml in pre and post-menopause respectively, and both groups of women had sedentary life style. Their receptor status especially of progesterone, estrogen and HER-2/neu were positive in 50% of premenopausal women and 65% of postmenopausal women. Conclusion: There are multiple physiological factors promoting breast cancer. High serum CA 15-3 level and hormonal imbalance of ER, PR and Her2/neu appears to be the main cause of breast cancer. It may be possible that the functional sites of these proteins may be altered which may increase the chances of metastasis in breast cancer.

THE EFFECT OF SEVERAL ROOT-END FILLING MATERIALS ON MG63 OSTEOBLAST-LIKE CELLS (수종의 치근단역충전 재료가 MG63 osteoblast-like cells에 미치는 영향)

  • Lee, Jeong-Ho;Shon, Won-Jun;Lee, Woo-Cheol;Baek, Seung-Ho
    • Restorative Dentistry and Endodontics
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    • v.35 no.3
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    • pp.222-228
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    • 2010
  • The purpose of this study was to compare mineral trioxide aggregate (MTA; Dentsply, Tulsa Dental, Tulsa, OK, USA), which is widely used as root-end filling material, with DiaRoot BioAggregate (DB; Innovative BioCaramix Inc, Vancouver, BC, Canada), newly developed product, by using MG63 osteoblast-like cells. MTA, DB, and Intermediate Restorative Material (IRM; Dentsply Caulk, Milford, DE, USA) were used for root-end filling material while tissue culture plastic was used for control group. Each material was mixed and, the mixtures were left to set for 24 hours. MG63 cells were seeded to each group and then they were cultured for attachment for 4 hours. Following the attachment of cells to the root-end filling material, early cellular response was observed. After another 12 hours'culture, the level of attachment between cells and material was observed and in order to identify the effect of each material to bone formation, transforming growth factor beta1 ($TGF{\beta}1$) and osteocalin (OC) were estimated by using enzyme-linked immunosorbent assay (ELISA), and the amount of alkaline phosphatase (ALP) was also measured. The data were analyzed using one-way ANOVA. As a result, only at OC and the number of cells which were attached to materials, there was no statistical difference between MTA and DB. At other items, there was statistically significant difference in all groups. Although DB has not shown exactly the same cellular response like that of MTA, the number of attached cells shows that biocompatibility of the material and OC indicates bone formation rate. Therefore, if DB is used for root end filling material, it is expected to lead to similar results to MTA.

Wound Healing Effects of Rose Placenta in a Mouse Model of Full-Thickness Wounds

  • Kim, Yang Woo;Baek, Seung Ryeol;Lee, Eun Sook;Lee, Sang Ho;Moh, Sang Hyun;Kim, Soo Yun;Moh, Ji Hong;Kondo, Chieko;Cheon, Young Woo
    • Archives of Plastic Surgery
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    • v.42 no.6
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    • pp.686-694
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    • 2015
  • Background Rosa damascena, a type of herb, has been used for wound healing in Eastern folk medicine. The goal of this study was to evaluate the effectiveness of rose placenta from R. damascena in a full-thickness wound model in mice. Methods Sixty six-week-old C57BL/6N mice were used. Full-thickness wounds were made with an 8-mm diameter punch. Two wounds were made on each side of the back, and wounds were assigned randomly to the control and experimental groups. Rose placenta ($250{\mu}g$) was injected in the experimental group, and normal saline was injected in the control group. Wound sizes were measured with digital photography, and specimens were harvested. Immunohistochemical staining was performed to assess the expression of epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), transforming growth factor-${\beta}1$ (TGF-${\beta}1$), and CD31. Vessel density was measured. Quantitative analysis using an enzyme-linked immunosorbent assay (ELISA) for EGF was performed. All evaluations were performed on postoperative days 0, 2, 4, 7, and 10. Statistical analyses were performed using the paired t-test. Results On days 4, 7, and 10, the wounds treated with rose placenta were significantly smaller. On day 2, VEGF and EGF expression increased in the experimental group. On days 7 and 10, TGF-${\beta}1$ expression decreased in the experimental group. On day 10, vessel density increased in the experimental group. The increase in EGF on day 2 was confirmed with ELISA. Conclusions Rose placenta was found to be associated with improved wound healing in a mouse full-thickness wound model via increased EGF release. Rose placenta may potentially be a novel drug candidate for enhancing wound healing.

The prevalence of pullorum disease-fowl typhoid in grand parent stock and parent stock in Korea, 2003 (2003년 국내 원종계 및 종계의 추백리-가금티푸스 감염실태)

  • Kim, Ae Ran;Kim, Jae Hong;Lee, Young Ju;Cho, Young Mi;Kwon, Jun Hun;Kwon, Yong Kuk;Lee, Youn Jeong;Choi, Jun Gu;Joh, Sung Jun;Kim, Min Chul;Lee, Eun Kyoung;Kim, Chang Seub;Yang, Hung Gu;Kwag, Sang Ick;Sung, Haan Woo;Moo, In Pil
    • Korean Journal of Veterinary Research
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    • v.46 no.4
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    • pp.347-353
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    • 2006
  • Serum samples of 30 chickens per flock from 6 grand parent stock (GPS) farms and 70 parent stock (PS) farms were collected for seroprevalent study of pullorum disease-fowl typhoid (PD-FT) infection by serum plate agglutination test (SPA). The incidence of PD-FT infection in GPS flocks and PS flocks were 0% and 15.7%, respectively. Especially PS flocks infected with PD-FT showed age dependent patterns that 22.2% of flocks between 20 to 30 weeks of age and 38.9% of flocks between 30 to 40 weeks of age were positive. The incidence of GPS flocks and PS flocks using Salmonella (S.) gallinarum 9R (SG9R) live vaccine were 33.3% and 58.6%, respectively. The sero-positive rate of 11 flocks were 6.7-83.3% by SPA and 2.9-55.6% by enzyme linked immunosorbent assay (ELISA), and ELISA showed more lower antibody levels than SPA. Furthermore, specific antibodies produced by SG9R vaccination were detectable by SPA using SG9R antigen without cross-reaction with the PD-FT infection.

Usefulness of the Neutrophil Gelatinase-Associated Lipocalin (NGAL) Kit for Acute Kidney Injury Patients at the Emergency Medical Center in Daegu (대구지역 응급의료센터에 내원한 급성 콩팥손상 환자의 진단을 위한 호중구 젤라티나제 관련 리포칼린 키트의 유용성)

  • Lee, Seung-Jin;Park, Sangwook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.49-53
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    • 2016
  • Acute kidney injury (AKI) is a common syndrome resulting in kidney damage and malfunction within a few days or even a few hours. The diagnosis of AKI depends on routine biochemical tests, including serum creatinine, aspartate aminotransferase (AST), alanine aminotransaminase (ALT), blood urea nitrogen (BUN), and electrolytes. Plasma neutrophil gelatinase-associated lipocalin (NGAL) is a biomarker that shows correlation with the severity of acute infections and kidney injuries. The predictive value in other conventional assays for kidney functions has been reported to cause distraction for AKI syndrome. The aim of this study is to verify the predictive value of plasma NGAL in patients with established AKI. The NGAL kit for checkup demonstrates sensitivity of ${\geq}300$ (92.2%), ${\geq}200$ (95.6%), ${\geq}100$ (99.6%), specificity of ${\geq}300$ (95.1%), ${\geq}200$ (97.3%), ${\geq}100$ (99.4%), positive predictability of ${\geq}300$ (93.3%), ${\geq}200$ (93.4%), ${\geq}100$ (99.2%), and negative predictability of ${\geq}300$ (96.7%), ${\geq}200$ (97.7%), ${\geq}100$ (98.1%), respectively. The plasma NGAL compared with the enzyme-linked immunosorbent assay (ELISA) has been shown to be an early predictive biomarker of AKI. The NGAL kit, recently developed for point-of-care of plasma specimens, is thought to be a useful and reliable biomarker for the early diagnosis of decreased kidney functions.

Immunoassay of Pertussis According to Ages (백일해에 대한 연령별 면역혈청학적 연구)

  • Lee, Soo Young;Choi, Ui Yoon;Kim, Ju Sang;Ahn, Joong Hyun;Choi, Jung Hyun;Ma, Sang Hyuk;Park, Joon Soo;Kim, Hwang Min;Kang, Jin Han
    • Pediatric Infection and Vaccine
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    • v.19 no.2
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    • pp.55-60
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    • 2012
  • Purpose: We conducted the immunoassay of pertussis according to ages, in order to evaluate protective immunity against pertussis in Korean populations. Methods: Healthy subjects were enrolled at four university hospitals in Korea. The subjects were grouped as seven age groups (every 10 years). Antibodies against pertussis toxin (PT) in sera were measured by enzyme linked immunosorbent assay (ELISA) kits. Geometric mean concentrations (GMC) of antibodies and the ratios of the subjects with seroprotective antibody levels were determined. The subjects with antibody titers ${\geq}24.0EU/mL$ were considered to seroprotective as the manufacturer's protocol. Results: Total 1,605 subjects (age: 2 months-65 years) participated in this study, and their GMC was $56.16{\pm}50.54EU/mL$. Among seven age groups, age group <11 year showed the highest GMC ($64.78{\pm}53.24EU/mL$) (P<0.001). In the analysis of the ratios of the subjects with seroprotective antibody titers, 68.2% of the subjects were proven to seroprotective, and age group <11 year also showed the highest ratio (76.5%) (P<0.001). Conclusions: We found that adolescences or adults (age group ${\geq}11$ year) showed lower levels of antibody against pertussis and lower ratio of the subjects with seroprotective antibody titers than children (age group <11 year).

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Study on Laboratory Diagnosis of the Ebola Virus and Its Current Trends (에볼라 바이러스 진단법과 개발 동향에 관한 고찰 연구)

  • Jeong, Hye Seon;Kang, Yun-Jung
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.105-111
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    • 2015
  • In late December 2013, the Ebola virus emerged from West Africa. The outbreak started in Guinea and rapidly spread to Liberia and Sierra Leone. Initially, the virus is spread to the human population after contact with infected wildlife and then spread person-to-person through direct contact with body fluids such as blood, sweat, urine, semen, and breast milk. The Ebola virus infects endothelial cells, mononuclear phagocytes and hepatocytes. It causes massive damage to internal tissues and organs, such as blood vessels and the liver, and ultimately death. Most tests for the virus RNA rely on a technology called reverse-transcriptase polymerase chain reaction (RT-PCR). While this method is highly sensitive, it is also expensive, requiring skilled scientists, and delicate power supplies. The strip analytical technique (enzyme-linked immunosorbent assay or ELISA) detects antigens or antibodies to the Ebola virus. This test is cheap and does not require electricity or refrigeration. Despite ongoing efforts directed at experimental treatments and vaccine development, current medical work on the Ebola viral disease is largely limited to supportive therapy. Thus, rapid and reliable diagnoses of the Ebola virus are critically important for patient management, infections, prevention, and control measures.

Effect of Piryongbanggamgil-tang on Airway Mucin Secretion, Production, Gene Expression and Hypersecretion of Mucus (필용방감길탕이 기도 뮤신의 분비, 생성, 유전자 발현 및 점액 과다 분비에 미치는 영향)

  • Kim, Yoon Young;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.28 no.2
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    • pp.56-71
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    • 2014
  • Objectives In this study, the author tried to investigate whether piryongbang-gamgil-tang (PGGT) significantly affect in vitro airway mucin secretion, PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production / gene expression from human airway epithelial cells and increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats. Materials and Methods For in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of PGGT to assess the effect of PGGT on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effect of PGGT on PMA- or EGFor TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PGGT and treated with PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24 hrs, to assess both effect of PGGT on PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production by ELISA and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). For in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to $SO_2$ during 3 weeks. Effect of orally-administered PGGT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assesed by using histopathological analysis after staining the epithelial tissue with alcian blue. Possible cytotoxicities of PGGT in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of PGGT were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering PGGT orally. Results (1) PGGT did not affect in vitro mucin secretion from cultured RTSE cells. (2) PGGT significantly inhibited PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. (3) PGGT decreased the amount of intraepithelial mucosubstances and showed the tendency of expectorating airway mucus already produced. (4) PGGT increased LDH release from RTSE cells. However, PGGT did not show in vivo liver and kidney toxicities and cytotoxicity to NCI-H292 cells. Conclusion The result from this study suggests that PGGT can regulate the production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of PGGT with their components should be further studied using animal experimental models that reflect the diverse pathophysiology of respiratory diseases through future investigations.

Study on the Changes in Allergen and Allergenicity Originated from Shrimp by Physical Treatments (물리적 처리에 의한 새우유래 Allergen 및 Allergenicity 변화)

  • Kim, Seong-Mi;Park, Jin-Gyu;Kim, Koth-Bong-Woo-Ri;Lee, Ju-Woon;Byun, Myung-Woo;Park, Sun-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.8
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    • pp.990-996
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    • 2006
  • This research was conducted to evaluate the changes in allergenicity of shrimp by physical treatments using competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). Shrimp was subjected to physical treatments such as high hydrostatic pressure (HHP), sonication, autoclave and microwave. Heat-stable protein (HSP) purified from raw shrimp was used as a major allergen. The binding ability of monoclonal IgG and shrimp-allergic patients' IgE to HSP treated with HHP decreased, increasing the pressure up to 400 MPa. Especially, it became less than 50% at 400 MPa. The binding ability of mAb to HSP treated with sonication (10, 20, 30 and 60 min) decreased with the treated time. Especially, it became less than 60% with the treatment for 60 min. The allergenicity change of HSP treated with autoclave and microwave little decreased. The binding ability of mAb to HSP during the treatment for 20 min became more than 70%. The results suggest that allergenicity of HSP in raw shrimp be more easily lost by HHP or sonication treatment than by autoclave or microwave treatment.