• 제목/요약/키워드: cELISA

검색결과 678건 처리시간 0.03초

Association of Viral Infections with Risk of Human Lymphomas, Egypt

  • Kadry, Dalia Y;Khorshed, Amira M;Rashed, Reham A;Mokhtar, Nadia M
    • Asian Pacific Journal of Cancer Prevention
    • /
    • 제17권4호
    • /
    • pp.1705-1712
    • /
    • 2016
  • Background: The aim of this study was to determine and evaluate the association of different viral infections, with hepatitis B and C viruses, Epstein-Barr virus, cytomegalovirus and human herpes virus-8 (HBV, HCV, EBV, CMV, HHV-8) with the risk of lymphomas (Hodgkin and non-Hodgkin) among Egyptian patients, and correlate with the histopathological staging and typing as well as the prevalence of combined infections. Materials and Methods: A total of 100 newly diagnosed lymphoma patients with 100 healthy age and sex matched normal controls were assayed for viral infection using enzyme linked immunosorbant assay (ELISA) followed by real time polymerase chain reaction (RT-PCR). Results: Our results showed a high statistical significant difference between cases and controls as regards clinical and laboratory findings (P<0.001 and=0.003). A high statistical difference was seen for the association of most viruses and lymphoma cases (p<0.001) except for positive HBs Ag, positive CMV IgG and HHV-8 (p=0.37, 0.70 and 1.0 respectively). No statistical significant difference was found between Hodgkin (HL) and non-Hodgkin (NHL) as regards viral prevalence except HCV antigen, 57.1% for HL and 26.5% for NHL (p = 0.03). Only, HBV DNA showed a high significant value among infiltrated bone marrow cases (p=0.003) and finally, a high significant association of 2 combined viral infections with infiltrated bone marrow lymphoma cases (p=0.04). Conclusions: Our results showed that infection with HBV, HCV, CMV and EBV were associated with increased risk of lymphoma among the Egyptian population. Detection of new associations between infectious agents and risk of cancer development will facilitate progress in elaboration of prophylactic measures, early diagnostic methods and, hopefully, novel therapy of malignant tumours.

One-Step Simultaneous Immunochromatographic Strip Test for Multianalysis of Ochratoxin A and Zearalenone

  • Shim, Won-Bo;Dzantiev, Boris B.;Eremin, Sergei A.;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
    • /
    • 제19권1호
    • /
    • pp.83-92
    • /
    • 2009
  • Individual immunochromatographic assays (ICG) for ochratoxin A (OTA) and zearalenone (ZEA) were optimized and used in the development of a one-step simultaneous immunochromatographic assay (OS-ICG) for the rapid multianalysis of two mycotoxins in corn samples. The nitrocellulose membrane of the OS-ICG was treated with OTA-bovine serum albumin (BSA), ZEA-ovalbumin (OVA), and anti-mouse IgG in the OTA test, ZEA test, and control zones, respectively. Monoclonal antibody-gold conjugates (OTA3 MAb-gold and ZEA2C5 MAb-gold) were sprayed onto the conjugate pad. The visual detection limits were 2.5 and 5 ng/ml for OTA and ZEA, respectively, and the results were obtained within 15 min after starting the analysis. An efficient, simple, and rapid extraction method using 30% MeOH/PBS was established and validated by analyzing the corn samples spiked with OTA/ZEA mixtures (0/0, 5/10, 10/20, and $20/30\;{\mu}g/kg$). The cut-off values of the OS-ICG for the spiked corn were 5 and $10\;{\mu}g/kg$ for OTA and ZEA, respectively. Natural corn samples were analyzed by OS-ICG, direct competitive enzyme-linked immunosorbent assay (DC-ELISA), and HPLC. Results of the OS-ICG were in good agreement with those obtained by DC-ELISA and HPLC. The developed OS-ICG offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site simultaneous determination of OTA and ZEA in cereals, food, and agricultural products in one analytical cycle.

IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen

  • Shin, Kwang-Hyun;Jeong, Kyoung-Yong;Hong, Chein-Soo;Yong, Tai-Soon
    • Parasites, Hosts and Diseases
    • /
    • 제47권1호
    • /
    • pp.31-36
    • /
    • 2009
  • Cockroaches have been recognized as a major cause of asthma. Bla g 4 is one of the most important German cockroach allergens. The aim of this study is to investigate IgE reactivity to the recombinant Bla g 4 (rBla g 4) in the sera of allergic patients and identify linear IgE binding epitope. For protein expression, full-length Bla g 4 (EF202172) was divided into 5 overlapping peptide fragments (E1: aa 1-100, E2: aa 34-77, E3: aa 74-117, E4: aa 114-156, and E5: aa 153-182). The full-length and 5 peptide fragments of Bla g 4 was generated by PCR and over-expressed in E. coli BL21 (DE3). The IgE binding reactivities of the full-length and peptide fragments were measured by ELISA using 32 serum samples of cockroach allergy. The sera of 8 patients (25%) reacted with rBla g 4. Four sera (100%) showed IgE-binding reactivity to full-length and peptide fragment 4, and 2 sera (50%) reacted with peptide fragment 2. One (20%) serum reacted with peptide fragment 3. The results of ELISA using overlapping recombinant fragments indicated that the epitope region was located at amino acid sequences 34-73 and 78-113, and major IgE epitope of Bla g 4 was located at amino acid sequences 118-152 of C-terminal. B-cell epitope analysis of German cockroach allergen Bla g 4 could contribute to the strategic development of more specific and potentially efficacious immunotherapy.

Serological Detection of Antibodies against Anaplasma spp. in Cattle Reared in the Gyeongsangbuk-do, Korea

  • Seo, Min-Goo;Ouh, In-Ouh;Lee, Seung-Hun;Son, Ui-Han;Geraldino, Paul John L.;Rhee, Man Hee;Kwon, Oh-Deog;Kim, Tae-Hwan;Kwak, Dongmi
    • Parasites, Hosts and Diseases
    • /
    • 제56권3호
    • /
    • pp.287-290
    • /
    • 2018
  • Anaplasmosis is a tick-borne, non-contagious, zoonotic disease caused by Anaplasma spp., which include Anaplasma marginale, A. centrale, A. phagocytophilum, A. platys, A. ovis, and A. bovis. Recently, in Korea, the prevalence of Anaplasma spp. has been investigated in some animals, such as dogs, horses, goats, cats, and Korean water deer. In cattle, A. marginale is the most virulent species and regarded as the typical type of species. However, data on the seroprevalence of Anaplasma spp. in cattle in Korea during the last decade is limited. This study was designed to investigate the seroprevalence of bovine anaplasmosis in Korea. From 2010 to 2013, blood samples were collected from 568 cattle. Forty animals (7.0%) tested seropositive for Anaplasma spp. by cELISA. Despite that current bovine anaplasmosis seropositivity rate in the Gyeongsangbuk-do is lower than those in tropical countries, anaplasmosis needs to be regarded as a concerning disease. The identification of the specific Anaplasma species infecting cattle in this province requires additional molecular studies. Moreover, further monitoring and control programs for bovine anaplasmosis is required, and the information from this study will be beneficial to develop these programs.

DA-9601, Artemisia Asiatica Herbal Extract, Ameliorates Airway Inflammation of Allergic Asthma in Mice

  • Kim, Ji Young;Kim, Dae Yong;Lee, Yun Song;Lee, Bong Ki;Lee, Kyung-Hoon;Ro, Jai Youl
    • Molecules and Cells
    • /
    • 제22권1호
    • /
    • pp.104-112
    • /
    • 2006
  • We previously reported that DA-9601, ethanol herbal extract of Artemisia asiatica, inhibited histamine and leukotriene releases in guinea pig lung mast cells activated with specific antigen/antibody reaction. This study aimed to evaluate the inhibitory effect of DA-9601 on the OVA-induced airway inflammation in allergic asthma mouse model. BALB/c mice were sensitized and challenged with OVA. DA-9601 was administered orally 1 h before every local OVA-challenge. OVA-specific serum IgE was measured by ELISA, recruitment of inflammatory cells in BAL fluids and lung tissues by Diff-Quik and H&E staining, respectively, the expressions of CD40, CD40L and VCAM-1 by immunohistochemistry, goblet cell hyperplasia by PAS staining, activities of MMPs by gelatin zymography, expressions of mRNA and proteins of cytokines by RT-PCR and ELISA, activities of MAP kinases by western blot, and activity of NF-${\kappa}B$ by EMSA. DA-9601 reduced IgE level, recruitment of inflammatory cells into the BAL fluid and lung tissues, expressions of CD40, CD40L and VCAM-1 molecules, goblet cell hyperplasia, MMPs activity, expressions of mRNA and productions of various cytokines, activities of MAP kinases and NK-${\kappa}B$ increased from OVA-challenged mice. These data suggest that DA-9601 may be developed as a clinical therapeutic agent in allergic diseases due to suppressing the airway allergic inflammation via regulation of various cellular molecules expressed by MAP kinases/NF-${\kappa}B$ pathway.

Three Predictive Tests Using Mice for the Identification of Contact Sensitizer

  • Jung-Hyun Shin;Min
    • 대한화장품학회지
    • /
    • 제22권2호
    • /
    • pp.201-210
    • /
    • 1996
  • Predictive tests for the identification of contact sensitizing chemicals have been developed. We measured the sensitization potential with three predictive tests, the in vitro and the in vivo Local Lymph Node Assay(LLNA), ELISA to detect interferon-gamma(IFN-${\gamma}$) from supernatant and flow cytometry to detect change of cell surface proteins, using draining lymph nodes of mice. BALB/c mice were exposed to various chemicals or vehicles on the ears daily for 3 consecutive days in all experiments. With some exceptions of propyl paraben, neomycin sulfate, the in vivo LLNA was able to detect the sensitizing capacity of test chemicals and was more sensitive than the in vitro LLNA for chemicals used in the present study. In another experiment, contact sensitivity was assessed by the ELISA to detect IFN-Υ from the supernatants of the cultured LNCs after sensitization with chemicals. There was a good correlation between the LLNA and the IFN-Υ production for test chemicals. We also examined the change of cell surface proteins on LNCs after sensitization by flow cytometry for some cell adhesion molecules(ICAM-1, E-cadherine, B7 molecule), T cell markers(CD3, CD4, CD8, T$\alpha$$\beta$,T${\gamma}$$\delta$) and B cell markers(LR1, CD45R, I-Ad). The number of ICAM-1 positive cells and B cells in LNCs were increased after sensitization with DNCB, TNCB, isoeugenol and 25%, 50% cinnamic aldehyde compared with that of vehicle as a control. In conclusion, the in vivo LLNA could provide more sensitive screening test for moderate to strong sensitizers and some weak sensitizers including cosmetic raw materials than the in vitro LLNA. The production of IFN-Υ by allergen-activated LNCs might be a values indicators without radioisotopes for the identification of contact allergens. Detection of allergens by testing the increase of ICAM-1 positive cells and B cells in LNCs by flow cytometry might be used as a test method to detect allergens.

  • PDF

The effect of plant extracts on the activity and the expression of MMPs (matrix metalloprotease) induced by UVA

  • Lee, Dong-hwan;Lee, Bum-chun;Yoon, Eun-jeong;Lee, Kyung-eun;Park, Sung-min;Pyo, Hyeong-bae;Choe, Tae-boo
    • 대한화장품학회:학술대회논문집
    • /
    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
    • /
    • pp.32-43
    • /
    • 2003
  • UV irradiation on a skin brings about the qualitative and quantitative alterations of the extracellular matrix. Repeated-UV irradiation suppressed the synthesis of collagen and activated the expression of the matrix metalloprotease (MMP). In this paper, on the purpose of development of novel anti-aging agents from natural sources, effects of several natural products on in vitro MMP-1 activity and UVA induced MMP-1 synthesis in human dermal fibroblast (HDF) culture were studied. We measured MMP-1 activities by fluorescence assay using gelatin as substrates. As a result, the extract of Dicentra spectabilis, and flower buds of Tussilago farfara showed strong inhibitory effect. Among them, the extract of flower buds of Tussilago fartara and Dicentra spectabilis inhibited MMP-1 activity by 92% and 87% at 0.05% (w/v). And UVA induced MMP-1 expression were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in HDF culture. The extract of flower buds of Tussilago farfara and Dicentra spectabilis suppressed the UVA induced expression of MMP-1 by similar level of Vitamin C 200$\mu$M at 0.1% (w/v). These results suggest that the extract of Dicentra spectabilis, and flower buds of Tussilago farfara effectively prevent skin from the UV-induced photoaging. So the extracts are thought to have potential as effective raw materials for anti-aging cosmetics.

  • PDF

Biocompatibility of two experimental scaffolds for regenerative endodontics

  • Leong, Dephne Jack Xin;Setzer, Frank C.;Trope, Martin;Karabucak, Bekir
    • Restorative Dentistry and Endodontics
    • /
    • 제41권2호
    • /
    • pp.98-105
    • /
    • 2016
  • Objectives: The biocompatibility of two experimental scaffolds for potential use in revascularization or pulp regeneration was evaluated. Materials and Methods: One resilient lyophilized collagen scaffold (COLL), releasing metronidazole and clindamycin, was compared to an experimental injectable poly(lactic-co-glycolic) acid scaffold (PLGA), releasing clindamycin. Human dental pulp stem cells (hDPSCs) were seeded at densities of $1.0{\times}10^4$, $2.5{\times}10^4$, and $5.0{\times}10^4$. The cells were investigated by light microscopy (cell morphology), MTT assay (cell proliferation) and a cytokine (IL-8) ELISA test (biocompatibility). Results: Under microscope, the morphology of cells coincubated for 7 days with the scaffolds appeared healthy with COLL. Cells in contact with PLGA showed signs of degeneration and apoptosis. MTT assay showed that at $5.0{\times}10^4$ hDPSCs, COLL demonstrated significantly higher cell proliferation rates than cells in media only (control, p < 0.01) or cells co-incubated with PLGA (p < 0.01). In ELISA test, no significant differences were observed between cells with media only and COLL at 1, 3, and 6 days. Cells incubated with PLGA expressed significantly higher IL-8 than the control at all time points (p < 0.01) and compared to COLL after 1 and 3 days (p < 0.01). Conclusions: The COLL showed superior biocompatibility and thus may be suitable for endodontic regeneration purposes.

Changes of cytokine mRNA expression and IgG responses In rats Infected with Capillaria hepatica

  • Kim, Dong-Kwan;Joo, Kyoung-Hwan;Chung, Myung-Sook
    • Parasites, Hosts and Diseases
    • /
    • 제45권2호
    • /
    • pp.95-102
    • /
    • 2007
  • The mRNA expression of several cytokines was evaluated in splenocytes and mesenteric lymph node (MLN) cells of rats infected with Capillaria hepatica by reverse-transcription (RT)-PCR until week 12 after infection. IgG1 and IgG2a, which are associated with Th1 and Th2 response, respectively, were also assessed by ELISA. The results indicated that the majority of cytokines, including the Th1 (IL-2 and $IFN-{\gamma}$ and Th2 cytokines (IL-4, IL-5 and IL-10) were expressed at maximal levels during the early stage of infection (after week 1-2), and the ELISA data also evidenced a similar pattern of changes in IgG1 arid IgG2a. Th1 and Th2 cytokines responded in a similar fashion in this rat model. The expression of cytokines in splenocytes was significantly higher than that in MLN cells, thereby indicating that cytokine production is controlled more by spleen than by MLN. In addition, the observation that $IFN-{\gamma}$ expression increased unexpectedly at the time of maximal egg production (6 weeks after infection) indicated that $IFN-{\gamma}$ is a cytokine reacting against egg production. However, increased IL-5 expression occurring in tandem with worm activity indicated that the activity of C. hepatica might be controlled by IL-5 expression.

만성폐쇄성폐질환 동물모델에서 SGX01의 폐손상 억제 효과 (Inhibitory Effects of SGX01 on Lung Injury of COPD Mice Model)

  • 박재준;양원경;유이란;김승형;박양춘
    • 대한한방내과학회지
    • /
    • 제40권4호
    • /
    • pp.567-581
    • /
    • 2019
  • Objective: This study aimed to evaluate the inhibitory effects of SGX01 on the lung injuries of COPD mice model. Materials and Methods: This study was carried out in two ways: in vitro and in vivo. In vitro, L929 cells were challenged with LPS, and then treated with six concentrations of SGX01 (10, 30, 50, 100, 300, and $500{\mu}g/ml$) and analyzed by ELISA. In vivo, C57BL/6 mice were challenged with LPS and cigarette smoking solution (CSS), and then treated with a vehicle only (control group), dexamethasone 3 mg/kg (dexa group), or a SGX01 200 mg/kg (SGX01 group). After sacrifice, the BALF or lung tissue was analyzed with Cytospin, FACS, ELISA, real-time PCR and H&E, and Masson's trichrome staining. Results: SGX01 significantly decreased NO, $TNF-{\alpha}$, and IL-6 on L929 cells challenged with LPS. In the COPD model, SGX01 significantly inhibited the increase of neutrophils, $TNF-{\alpha}$, IL-17A, CXCL-1, MIP2, CD8+ cells in BALF, and $TNF-{\alpha}$, $IL-1{\beta}$ mRNA expression in lung tissue. It also decreased the severity of the histological lung injury. Conclusion: This study suggests the usability of SGX01 for COPD patients by controlling lung tissue injury.